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Sample records for abortus strains isolated

  1. Identifikasi Brucella abortus Isolat Lokal dengan Brucella abortus Strain Specific-Polymerase Chain Reaction (IDENTIFICATION OF LOCAL ISOLATES OF BRUCELLA ABORTUS USING BRUCELLA ABORTUS STRAIN SPECIFIC-POLYMERASE CHAIN REACTION ASSAY

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    Susan Maphilindawati Noor

    2014-10-01

    Full Text Available Brucella abortus Strain Specific-Polymerase Chain Reaction (BaSS-PCR is a single multiplex PCRtechnique which able to identify and differentiate between Brucella abortus field strains (biovar 1, 2, and4, B. abortus vaccine strains, Brucella species, and non-Brucella species. In this study, BaSS-PCR wasapplied to identify local isolates of B. abortus in order to investigate the B. abortus strains that infectedcattle in Indonesia. Fifty local strains of B.abortus isolated from infected cattle in Java (Jakarta andBandung, South Sulawesi (Maros, East Nusa Tenggara (Kupang and Belu were used in this study. TheDNA bands were observed by agarose gel in the presence of ethidium bromide. Identification was performedbased on the size and number of DNA products amplified by PCR from each isolates. The results showedthat the 50 isolates were of B. abortus field strains. This finding showed that the cause of bovine brucellosisin Indonesia is B. abortus field strains.

  2. Use of polymerase chain reaction to identify Brucella abortus strain RB51 among Brucella field isolates from cattle in Italy.

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    Adone, R; Ciuchini, F; La Rosa, G; Marianelli, C; Muscillo, M

    2001-03-01

    Brucella abortus strain RB51, a rough mutant of the B. abortus 2308 virulent strain, was recently approved in the United States as the official vaccine for brucellosis in cattle. Following recent evidence of unauthorized use of RB51 vaccine in Italy, where the use of vaccines for brucellosis is no longer allowed, the suitability of an RB51-specific polymerase chain reaction assay for identifying the RB51 strain among Brucella field isolates from cattle in Italy was investigated. The oligonucleotide primers used in this study, belonging to a six-primer cocktail for Brucella species previously described by other authors, allowed the amplification of a 364-base pair (bp) fragment specific for RB51 and its parent strain 2308, and a 498-bp product specific for B. abortus. In addition, unresolved bands ranging from 600 to 700 bp were observed from RB51 strain. Brucella abortus biovars 1, 2 and 4 have only one specific sensitive 498-bp band. The B. abortus biovars 3, 5 and 6 did not give any signal. The 498-bp product from a reference Brucella strain was sequenced and submitted to EMBL with the accession number AJ271969 while the 364-bp fragment from RB51 strain was submitted to EMBL database with accession number AJ271968. The sequence studies confirmed the specificity of the detected fragments. No amplification was obtained by testing DNA from strains antigenically related to Brucella, such as Yersinia enterocolitica O:9, Escherichia coli O:157, Salmonella urbana and Pasteurella multocida. The results of this study indicate that this technique, in combination with specific serological tests, could be a useful diagnostic method to verify the use of RB51 vaccine and can contribute to the creation of a databank of circulating strains.

  3. Dissemination and genetic diversity of chlamydial agents in Polish wildfowl: Isolation and molecular characterisation of avian Chlamydia abortus strains.

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    Monika Szymańska-Czerwińska

    Full Text Available Wild birds are considered as a reservoir for avian chlamydiosis posing a potential infectious threat to domestic poultry and humans. Analysis of 894 cloacal or fecal swabs from free-living birds in Poland revealed an overall Chlamydiaceae prevalence of 14.8% (n = 132 with the highest prevalence noted in Anatidae (19.7% and Corvidae (13.4%. Further testing conducted with species-specific real-time PCR showed that 65 samples (49.2% were positive for C. psittaci whereas only one was positive for C. avium. To classify the non-identified chlamydial agents and to genotype the C. psittaci and C. avium-positive samples, specimens were subjected to ompA-PCR and sequencing (n = 83. The ompA-based NJ dendrogram revealed that only 23 out of 83 sequences were assigned to C. psittaci, in particular to four clades representing the previously described C. psittaci genotypes B, C, Mat116 and 1V. Whereas the 59 remaining sequences were assigned to two new clades named G1 and G2, each one including sequences recently obtained from chlamydiae detected in Swedish wetland birds. G1 (18 samples from Anatidae and Rallidae grouped closely together with genotype 1V and in relative proximity to several C. abortus isolates, and G2 (41 samples from Anatidae and Corvidae grouped closely to C. psittaci strains of the classical ABE cluster, Matt116 and M56. Finally, deep molecular analysis of four representative isolates of genotypes 1V, G1 and G2 based on 16S rRNA, IGS and partial 23S rRNA sequences as well as MLST clearly classify these isolates within the C. abortus species. Consequently, we propose an expansion of the C. abortus species to include not only the classical isolates of mammalian origin, but also avian isolates so far referred to as atypical C. psittaci or C. psittaci/C. abortus intermediates.

  4. Molecular characterisation and ovine live vaccine 1B evaluation toward a Chlamydophila abortus strain isolated from springbok antelope abortion.

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    Berri, M; Bernard, F; Lecu, A; Ollivet-Courtois, F; Rodolakis, A

    2004-11-15

    Chlamydiosis is a zoonosis with a worldwide distribution. The reservoir of susceptible hosts is large and includes birds and both domestic and wild mammals. Chlamydial infection, determined serologically, seems to be widespread among wild ruminants in the Paris zoo (France). In February 2003, an abortion case was reported within the springbok (Antidorcas marsupialis) herd of the zoo. PCR assay using primers targeting the polymorph membrane protein gene (pmp) family was performed on both vaginal swab and placenta samples revealing the presence of Chlamydophila. The inoculation into chicken embryos of an infected placenta extract led to the successful isolation of a C. abortus strain referred to as ASb1. The omp1 gene coding the major outer membrane protein (momp) and the 16S-23S rRNA spacer region of ASb1 were compared to those of various strains by restriction fragment length polymorphism (RFLP). The RFLP analysis showed that this isolate belonged to Chlamydophila abortus species and is highly related to known domestic ruminant's strains causing abortion. The efficacy of a live vaccine 1B, based on a temperature-sensitive mutant of the ovine abortion reference strain AB7, was tested. Protection-challenge experiments in a mouse model show that the ASb1 strain led to mice abortions and that vaccination with 1B vaccine provided them with effective protection.

  5. Exploring the Diversity of Field Strains of Brucella abortus Biovar 3 Isolated in West Africa

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    Moussa Sanogo

    2017-06-01

    Full Text Available Brucellosis is one of the most widespread bacterial zoonotic diseases in the world, affecting both humans and domestic and wild animals. Identification and biotyping of field strains of Brucella are of key importance for a better knowledge of the epidemiology of brucellosis, for identifying appropriate antigens, for managing disease outbreaks and for setting up efficient preventive and control programmes. Such data are required both at national and regional level to assess potential threats for public health. Highly discriminative genotyping methods such as the multiple locus variable number of tandem repeats analysis (MLVA allow the comparison and assessment of genetic relatedness between field strains of Brucella within the same geographical area. In this study, MLVA biotyping data retrieved from the literature using a systematic review were compared using a clustering analysis and the Hunter-Gaston diversity index (HGDI. Thus, the analysis of the 42 MLVA genotyping results found in the literature on West Africa [i.e., from Ivory Coast (1, Niger (1, Nigeria (34, The Gambia (3, and Togo (3] did not allow a complete assessment of the actual diversity among field strains of Brucella. However, it provided some preliminary indications on the co-existence of 25 distinct genotypes of Brucella abortus biovar 3 in this region with 19 genotypes from Nigeria, three from Togo and one from Ivory Coast, The Gambia, and Niger. The strong and urgent need for more sustainable molecular data on prevailing strains of Brucella in this sub-region of Africa and also on all susceptible species including humans is therefore highlighted. This remains a necessary stage to allow a comprehensive understanding of the relatedness between field strains of Brucella and the epidemiology of brucellosis within West Africa countries.

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

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    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.

  7. Molecular strain typing of Brucella abortus isolates from Italy by two VNTR allele sizing technologies.

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    De Santis, Riccardo; Ancora, Massimo; De Massis, Fabrizio; Ciammaruconi, Andrea; Zilli, Katiuscia; Di Giannatale, Elisabetta; Pittiglio, Valentina; Fillo, Silvia; Lista, Florigio

    2013-10-01

    Brucellosis, one of the most important re-emerging zoonoses in many countries, is caused by bacteria belonging to the genus Brucella. Furthermore these bacteria represent potential biological warfare agents and the identification of species and biovars of field strains may be crucial for tracing back source of infection, allowing to discriminate naturally occurring outbreaks instead of bioterrorist events. In the last years, multiple-locus variable-number tandem repeat analysis (MLVA) has been proposed as complement of the classical biotyping methods and it has been applied for genotyping large collections of Brucella spp. At present, the MLVA band profiles may be resolved by automated or manual procedures. The Lab on a chip technology represents a valid alternative to standard genotyping techniques (as agarose gel electrophoresis) and it has been previously used for Brucella genotyping. Recently, a new high-throughput genotyping analysis system based on capillary gel electrophoresis, the QIAxcel, has been described. The aim of the study was to evaluate the ability of two DNA sizing equipments, the QIAxcel System and the Lab chip GX, to correctly call alleles at the sixteen loci including one frequently used MLVA assay for Brucella genotyping. The results confirmed that these technologies represent a meaningful advancement in high-throughput Brucella genotyping. Considering the accuracy required to confidently resolve loci discrimination, QIAxcel shows a better ability to measure VNTR allele sizes compared to LabChip GX.

  8. Evaluation of the HOOF-Print assay for typing Brucella abortus strains isolated from cattle in the United States: results with four performance criteria

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    Ewalt Darla R

    2005-06-01

    Full Text Available Abstract Background A fundamental question that arises during epidemiological investigations of bacterial disease outbreaks is whether the outbreak strain is genetically related to a proposed index strain. Highly discriminating genetic markers for characterizing bacterial strains can help in clarifying the genetic relationships among strains. Under the auspices of the European Society of Clinical Microbiology and Infectious Diseases, the European Study Group for Epidemiological Markers (ESGEM established guidelines for evaluating the performance of typing systems based of a number of criteria. Recently, HOOF-Print genotype analysis, a new method for typing Brucella abortus strains based on hypervariability at eight tandem repeat loci, was described. This paper evaluates the HOOF-Print assay by four of the criteria set out by the ESGEM: typeability, reproducibility, power of discrimination, and concordance with other typing methods. Results The HOOF-Print Assay was evaluated with a test population composed of 97 unrelated field isolates and 6 common laboratory strains of B. abortus. Both typeability and reproducibility of the assay were excellent. Allele diversity and frequency varied widely among the eight loci, ranging from 1 to 13 alleles. The power of discrimination, measured by the Hunter-Gaston discrimination index (HGDI, varied by locus ranging from 0 to 0.89, where a maximal value of 1.0 indicates discrimination of all strains. The HGDI values calculated for subgroups sorted by biovar were similar to the values determined for the whole population. None of the individual loci achieved the recommended HGDI threshold of 0.95, but the HGDI of the composite profiles was 0.99 (93 unique genotypes from 97 field strains evaluated, well above the recommended threshold. By comparison, the HGDI value for biovar typing was 0.61 in a test population biased with disproportionate numbers of the less common biovars. Cluster analysis based on HOOF

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

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    Chauhan, H C; Patel, Bharat K; 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

    2016-10-27

    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. Copyright © 2016 Chauhan et al.

  10. Brucella abortus RB51 induces protection in mice orally infected with the virulent strain B. abortus 2308.

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    Pasquali, Paolo; Rosanna, Adone; Pistoia, Claudia; Petrucci, Paola; Ciuchini, Franco

    2003-05-01

    Brucellae are gram-negative, facultative intracellular bacteria which are one of the most common causes of abortion in animals. In addition, they are the source of a severe zoonosis. In this trial, we evaluated the effect of oral inoculation of Brucella abortus RB51 in mice against a challenge infection with B. abortus 2308. First, we showed that a gastric acid neutralization prior to the oral inoculation contributed to a more homogeneous and consistent infection with both vaccine strain B. abortus RB51 and virulent strain B. abortus 2308. Successively, we assessed the clearance and the immune response following an oral infection with B. abortus RB51. Oral inoculation gave a mild infection which was cleared 42 days after infection, and it induced a delayed humoral and cell-mediated immune response. Finally, we immunized mice by oral inoculation with B. abortus RB51, and we challenged them with the virulent strain B. abortus 2308 by an oral or intraperitoneal route 42 days after vaccination. Oral inoculation of B. abortus RB51 was able to give protection to mice infected with the virulent strain B. abortus 2308 by the oral route but not to mice infected intraperitoneally. Our results indicate that oral inoculation of mice with B. abortus RB51 is able to give a protective immunity against an oral infection with virulent strains, and this protection seems to rely on an immune response at the mucosal level.

  11. 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.

  12. Post-exposure serological and bacteriological responses of water buffalo (Bubalus bubalis) to Brucella abortus biovar 1 following vaccination with Brucella abortus strain RB51.

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    Diptee, M D; Asgarali, Z; Campbell, M; Fosgate, G; Adesiyun, A A

    2007-12-01

    Serological and bacteriological responses to Brucella abortus biovar 1 following vaccination with B. abortus strain RB51 (RB51) were evaluated in thirty domestic water buffalo (Bubalus bubalis) randomly divided into five treatment groups. Groups I to V received, respectively, the recommended dose (RD) of RB51 vaccine once, RD twice 4 weeks apart, double RD once, double RD twice 4 weeks apart, and saline once (control). Vaccination did not result in a serological response. Experimental animals released 27 weeks post initial inoculation (27 PIIW) into a brucellosis-positive herd failed to seroconvert after 29 weeks. Experimental challenge commenced at 57 PIIW. All animals received B. abortus biovar 1 intraconjunctivally at 0, 5 and 9 weeks post experimental exposure (PEEW). Serum samples collected at 4, 8 and 13 PEEW were negative. At 16 PEEW all animals received B. abortus biovar 1 subcutaneously (SC), and all seroconverted by 20 PEEW. Five of twenty-six animals were positive for Brucella infection on bacterial culture. Brucella abortus biovar 1 was isolated from three animals; B. abortus RB51 was isolated from two. Treatment group, age and sex had no effect on the isolation of Brucellae (P>0.05).

  13. Molecular epidemiology of Brucella abortus isolated from cattle in Brazil, 2009-2013.

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    Oliveira, Mayra Silva; Dorneles, Elaine Maria Seles; Soares, Paulo Martins Filho; Fonseca, Antônio Augusto; Orzil, Lívia; de Souza, Patrícia Gomes; Lage, Andrey Pereira

    2017-02-01

    The aims of the present study were to genotype Brucella abortus strains isolated from cattle in Brazil between 2009 and 2013, and to analyze their distribution to support the Programa Nacional de Controle e Erradicação de Brucelose e Tuberculose (PNCEBT) (National Brucellosis and Tuberculosis Control and Eradication Program). One hundred forty B. abortus strains isolated from cattle in Brazil between 2009 and 2013 were genotyped using a set of 18 variable number of tandem repeats (VNTR) (MLVA16+HOOF-Print 3 and 4). The multiple locus VNTR analysis (MLVA) composed by eight markers (MLVA8) revealed eight different genotypes among B. abortus strains, including five previously described and three new ones. Analysis of the MLVA16 loci revealed fifty-eight distinct genotypes, from which three were identical, thirty-eight were considered very close, and seventeen were considered distant compared to those previously described and deposited in MLVAbank. Analysis of the HOOF-Prints 3 and 4 revealed the larger number of different alleles among all VNTR assessed, exhibiting maximum resolution when associated with MLVA16 markers. This study also provides insights on the genotypes of B. abortus circulating in Brazil, which certainly contribute for the better understanding of the epidemiology and control of bovine brucellosis in the country. Moreover, our data showed a high genetic diversity among the B. abortus strains isolated between 2009 and 2013, and a close relationship among these strains and Brazilian B. abortus deposited by MLVAbank. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Characterization of genomic island 3 and genetic variability of Chilean field strains of Brucella abortus.

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    Céspedes, Sandra; Salgado, Paulina; Valenzuela, Patricio; Vidal, Roberto; Oñate, Angel A

    2011-07-01

    One of the capabilities developed by bacteria is the ability to gain large fragments of DNA from other bacteria or to lose portions of their own genomes. Among these exchangeable fragments are the genomic islands (GIs). Nine GIs have been identified in Brucella, and genomic island 3 (GI-3) is shared by two pathogenic species, B. melitensis and B. abortus. GI-3 encodes mostly unknown proteins. One of the aims of this study was to perform pulsed-field gel electrophoresis (PFGE) on field isolates of B. abortus from Chile to determine whether these isolates are clonally related. Furthermore, we focused on the characterization of GI-3, studying its organization and the genetic conservation of the GI-3 sequence using techniques such as tiling-path PCR (TP-PCR) and restriction fragment length polymorphism-PCR (RFLP-PCR). Our results, after PFGE was performed on 69 field isolates of B. abortus from Chile, showed that the strains were genetically homogeneous. To increase the power of genetic discrimination among these strains, we used multiple locus variable-number tandem-repeat (VNTR) analysis with 16 loci (MLVA-16). The results obtained by MLVA-16 showed that the strains of B. abortus were genetically heterogeneous and that most of them clustered according to their geographic origin. Of the genetic loci studied, panel 2B was the one describing the highest diversity in the analysis, as well as locus Bruce19 in panel 2A. In relation to the study of GI-3, our experimental analysis by TP-PCR identified and confirmed that GI-3 is present in all wild strains of B. abortus, demonstrating the high stability of gene cluster GI-3 in Chilean field strains.

  15. Outer Membrane Proteins of Brucella abortus Vaccinal and Field Strains and their Immune Response in Buffaloes

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    Rukhshanda Munir*, M. Afzal1, M. Hussain2, S. M. S. Naqvi3 and A. Khanum3

    2010-04-01

    Full Text Available Outer membrane proteins (OMPs of three strains of B. abortus i.e. S19, RB51 and a local field isolate of biotype 1 were isolated through disrupting cells to generate membranes by centrifugation and sodium lauryl sarcosinate solubilisation of inner membrane proteins. Distinct OMP profiles of each strain were seen on SDS-PAGE. SDS-PAGE analysis of S19 and field isolate revealed eight protein bands in each strain. The OMPs of S19 had molecular masses 89.0, 73.0, 53.7, 49.0, 38.0, 27.0, 22.3, and 17.7 kDa, while field isolate had OMPs of 151.3, 89.0, 75.8, 67.6, 37.0, 27.0, 24.0 and 19.0 kDa. B. abortus RB51 yielded 11 OMP bands ranging from 12.5 to 107.1 kDa, with 34.2, 15.8 and 12.5 kDa as additional OMPs. Western immunoblot analysis using antisera raised against all three strains in buffaloes indicated an almost similar pattern of immuno-reactive OMPs in S19 and field strain. Two OMPs of molecular weight 37-38 and 19 kDa were immuno-reactive in all strains in buffaloes. There is possibility of use of these OMPs in a recombinant vaccine for B. abortus. A distinct protein of molecular weight of 151.3 kDa was identified in field strain but not in both vaccine strains of B. abortus. Use of this OMP in a diagnostic assay may differentiate between vaccinated and infected animals.

  16. Abortion and premature birth in cattle following vaccination with Brucella abortus strain RB51.

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    Fluegel Dougherty, Amanda M; Cornish, Todd E; O'Toole, Donal; Boerger-Fields, Amy M; Henderson, Owen L; Mills, Ken W

    2013-09-01

    Brucella abortus RB51 is the vaccine strain currently licensed for immunizing cattle against brucellosis in the United States. Most cattle are vaccinated as heifer calves at 4-12 months of age. Adult cattle may be vaccinated in selected high-risk situations. Two herds of pregnant adult cattle in the brucellosis-endemic area of Wyoming were vaccinated with a standard label dose (1.0-3.4 × 10(10) organisms) of RB51. Reproductive losses in the vaccinated herds were 5.3% (herd A) and 0.6% (herd B) and included abortions, stillbirths, premature calves, and unbred cows (presumed early abortion). Brucella abortus was cultured from multiple tissues of aborted and premature calves (7/9), and from placenta. Isolates were identified as B. abortus strain RB51 by standard strain typing procedures and a species-specific polymerase chain reaction. Bronchopneumonia with intralesional bacteria and placentitis were observed microscopically. There was no evidence of involvement of other infectious or toxic causes of abortion. Producers, veterinarians, and laboratory staff should be alert to the risk of abortion when pregnant cattle are vaccinated with RB51, to potential human exposure, and to the importance of distinguishing field from vaccinal strains of B. abortus.

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

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    Caporale, Vincenzo; Bonfini, Barbara; Di Giannatale, Elisabetta; Di Provvido, Andrea; Forcella, Simona; Giovannini, Armando; Tittarelli, Manuela; Scacchia, Massimo

    2010-01-01

    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.

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

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    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. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. A rapid minor groove binder PCR method for distinguishing the vaccine strain Brucella abortus 104M.

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    Nan, Wenlong; Qin, Lide; Wang, Yong; Zhang, Yueyong; Tan, Pengfei; Chen, Yuqi; Mao, Kairong; Chen, Yiping

    2018-01-24

    Brucellosis is a widespread zoonotic disease caused by Gram-negative Brucella bacteria. Immunisation with attenuated vaccine is an effective method of prevention, but it can interfere with diagnosis. Live, attenuated Brucella abortus strain 104M has been used for the prevention of human brucellosis in China since 1965. However, at present, no fast and reliable method exists that can distinguish this strain from field strains. Single nucleotide polymorphism (SNP)-based assays offer a new approach for such discrimination. SNP-based minor groove binder (MGB) and Cycleave assays have been used for rapid identification of four Brucella vaccine strains (B. abortus strains S19, A19 and RB51, and B. melitensis Rev1). The main objective of this study was to develop a PCR assay for rapid and specific detection of strain 104M. We developed a SNP-based MGB PCR assay that could successfully distinguish strain 104M from 18 representative strains of Brucella (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae, and B. ovis), four Brucella vaccine strains (A19, S19, S2, M5), and 55 Brucella clinical field strains. The assay gave a negative reaction with four non-Brucella species (Escherichia coli, Pasteurella multocida, Streptococcus suis and Pseudomonas aeruginosa). The minimum sensitivity of the assay, evaluated using 10-fold dilutions of chromosomal DNA, was 220 fg for the 104M strain and 76 fg for the single non-104M Brucella strain tested (B. abortus A19). The assay was also reproducible (intra- and inter-assay coefficients of variation = 0.006-0.022 and 0.012-0.044, respectively). A SNP-based MGB PCR assay was developed that could straightforwardly and unambiguously distinguish B. abortus vaccine strain 104M from non-104M Brucella strains. Compared to the classical isolation and identification approaches of bacteriology, this real-time PCR assay has substantial advantages in terms of

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

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    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.

  2. 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.

  3. 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.

  4. First isolation, identification, phenotypic and genotypic characterization of Brucella abortus biovar 3 from dairy cattle in Tanzania.

    Science.gov (United States)

    Mathew, C; Stokstad, M; Johansen, T B; Klevar, S; Mdegela, R H; Mwamengele, G; Michel, P; Escobar, L; Fretin, D; Godfroid, J

    2015-07-21

    Brucellosis is a disease of worldwide public health and economic importance. Successful control is based on knowledge of epidemiology and strains present in an area. In developing countries, most investigations are based on serological assays. This study aimed at investigating a dairy herd experiencing abortions in order to establish within-herd seroprevalence to Brucella spp., identify, characterize Brucella strains by Multiple Loci Variable Number of Tandem Repeats Analysis (MLVA-VNTR) and investigate possible spillover to other species. The within-herd seroprevalence in cattle (n = 200) was 48 % (95 % CI 41-55), using an indirect ELISA, while the Rose Bengal Test (RBT) yielded lower prevalence (21.5 %; 95 % CI 16-27). Two sheep (n = 35) and one goat (n = 50) were seropositive using ELISA while none of the dogs (n = 6) was positive with the RBT. Three Brucella were isolated from an aborted fetus and associated membranes. Real time PCR (IS711), Bruce-ladder and classical biotyping classified the isolates as B. abortus biovar 3. MLVA-VNTR revealed two different but closely related genotypes. The isolates showed unique profiles, providing the first genotypic data from Tanzania. These genotypes were not related to B. abortus biovar 3 reference strain Tulya originally isolated from a human patient in Uganda in 1958, unlike the genotypes isolated and characterized recently in Kenya. High within-herd prevalence, isolation of the pathogen and abortion confirm that B. abortus is circulating in this herd with cattle as reservoir hosts. A low seroprevalence in sheep and goats suggests a spillover of B. abortus from cattle to small ruminants in the herd. This is the first isolation and characterization of B. abortus biovar 3 from a dairy cow with abortion in Tanzania. The origin of the Tanzanian genotypes remain elusive, although they seem to be related to genotypes found in Europe, Turkey and China but not related to B. abortus biovar 3 reference strain or genotypes from

  5. Protein profile of Chlamydophila abortus isolates from Kerala, India

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    Binu K Mani

    Full Text Available Chlamydiae are of microbiological interest because of their mode of interaction with eukaryotic host cells and their specialized life cycle with unique features of parasitism. Reports regarding prevalence of infections of Chlamydophila abortus, the causative organism for chlamydial abortions in livestock, was the basis of the study. Two isolates, one each from cattle and goat abortion along with a reference isolate, were used for characterization with Sodium Dodecyl Sulphate-Poly Acrylamide Gel Electrophoresis (SDS-PAGE. Elementary bodies infected Mc Coy cells, harvested from bottle cultures were disrupted by Teflon coated magnetic pellet. Urografin-76 diluted with Tris-Potassium hydrochloride was used for purification of Elementary bodies of Chlamydophila abortus organism. On protein estimation of Elementary bodies by Biuret method, all the three isolates revealed protein concentration between 500-1000 mg/100ml, which were sufficient for electrophoresis. Ten percent of resolving gel and five percent of stacking gel of polyacrylamide in which 10g of processed isolate samples along with standard protein marker and Mc Coy cell protein (control were electrophoresed. Using Alpha Imager Gel Documentation System, the protein bands were analyzed. Twelve bands each for local bovine isolate and reference isolate were noticed while only 10 bands were there in the caprine isolate. Additional bands of 148 kDa and 135 kDa were present in bovine isolate, compared to the reference isolate, while 152 kDa and 137 kDa bands were unique for caprine isolate. [Vet. World 2011; 4(10.000: 470-472

  6. Safety of Brucella abortus strain RB51 in deer mice.

    Science.gov (United States)

    Cook, W E; Williams, E S; Thorne, E T; Taylor, S K; Anderson, S

    2001-07-01

    Brucella abortus strain RB51 is an approved brucellosis vaccine for use in cattle that may have potential as an oral vaccine for use in elk (Cervus elaphus) and/or bison (Bison bison). This study was designed to determine effects of strain RB51 on deer mice (Peromyscus maniculatus), a nontarget species that could have access to treated baits in a field situation. In February 1994, 90 mice were orally dosed or intraperitoneally injected with 1 x 10(8) colony forming units strain RB51 and 77 controls were similarly dosed with sterile saline. At weekly intervals through early April 1994, 4 to 6 mice from each group were euthanized, gross necropsies performed, spleens and uteruses cultured, and tissues examined histologically. All orally inoculated mice cleared the infection by 6 wk post-inoculation (PI). While most of the injected mice cleared the infection by 7 wk PI, a few required 9 wk. There were minimal adverse effects attributable to strain RB51. Apparently, strain RB51 would not negatively impact P. maniculatus populations if it were used in a field situation. Also, deer mice appear to be able to clear the vaccine in 6 to 9 wk, thus the probability of these mice transmitting the vaccine to other animals is low.

  7. Safety of B. abortus rough mutant strain RB51 administration in Buffalo cows

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    G. Iovane

    2010-02-01

    Full Text Available The objective of this study was to determine if B. abortus rough mutant strain RB51 is eliminated in Buffalo milk. Five milk buffaloes were inoculated with the triple of the recommended calfhood dose (3.0 – 10.2 x 1010 cfu/ml of B. abortus RB51 strain by subcutaneous route in the right axillary region. Milk samples were taken aseptically on a daily basis for the first 30 days and weekly for the second and third months. The samples were inoculated on Brucella Medium Base (BMB (Oxoid and Rifampin Brucellae Medium (RBM and incubated under 10% CO2 at 37°C for 10 days. The suspicious colonies were recultured in BMB and RBM. PCR analysis was also performed on milk samples. There were no isolations of bacteria with characteristics of Brucella from any of the milk samples collected during 90 days of the study. However Brucella RB51 DNA was detected on day 2 and 3 post vaccination in one buffalo cow and on day 21 post vaccination in another buffalo cow. It was concluded that the strain used at this dose wasn’t eliminated by milk in Buffaloes inoculated during lactation, however PCR positive results underline the necessity of milk pasteurization in order to minimize food-chain exposure.

  8. 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.

  9. Molecular epidemiology of Brucella abortus isolates from cattle, elk, and bison in the United States, 1998 to 2011.

    Science.gov (United States)

    Higgins, James; Stuber, Tod; Quance, Christine; Edwards, William H; Tiller, Rebekah V; Linfield, Tom; Rhyan, Jack; Berte, Angela; Harris, Beth

    2012-05-01

    A variable-number tandem repeat (VNTR) protocol targeting 10 loci in the Brucella abortus genome was used to assess genetic diversity among 366 field isolates recovered from cattle, bison, and elk in the Greater Yellowstone Area (GYA) and Texas during 1998 to 2011. Minimum spanning tree (MST) and unweighted-pair group method with arithmetic mean (UPGMA) analyses of VNTR data identified 237 different VNTR types, among which 14 prominent clusters of isolates could be identified. Cattle isolates from Texas segregated into three clusters: one comprised of field isolates from 1998 to 2005, one comprised of vaccination-associated infections, and one associated with an outbreak in Starr County in January 2011. An isolate obtained from a feral sow trapped on property adjacent to the Starr County herd in May 2011 clustered with the cattle isolates, suggesting a role for feral swine as B. abortus reservoirs in Starr County. Isolates from a 2005 cattle outbreak in Wyoming displayed VNTR-10 profiles matching those of strains recovered from Wyoming and Idaho elk. Additionally, isolates associated with cattle outbreaks in Idaho in 2002, Montana in 2008 and 2011, and Wyoming in 2010 primarily clustered with isolates recovered from GYA elk. This study indicates that elk play a predominant role in the transmission of B. abortus to cattle located in the GYA.

  10. Molecular Epidemiology of Brucella abortus Isolates from Cattle, Elk, and Bison in the United States, 1998 to 2011

    Science.gov (United States)

    Stuber, Tod; Quance, Christine; Edwards, William H.; Tiller, Rebekah V.; Linfield, Tom; Rhyan, Jack; Berte, Angela; Harris, Beth

    2012-01-01

    A variable-number tandem repeat (VNTR) protocol targeting 10 loci in the Brucella abortus genome was used to assess genetic diversity among 366 field isolates recovered from cattle, bison, and elk in the Greater Yellowstone Area (GYA) and Texas during 1998 to 2011. Minimum spanning tree (MST) and unweighted-pair group method with arithmetic mean (UPGMA) analyses of VNTR data identified 237 different VNTR types, among which 14 prominent clusters of isolates could be identified. Cattle isolates from Texas segregated into three clusters: one comprised of field isolates from 1998 to 2005, one comprised of vaccination-associated infections, and one associated with an outbreak in Starr County in January 2011. An isolate obtained from a feral sow trapped on property adjacent to the Starr County herd in May 2011 clustered with the cattle isolates, suggesting a role for feral swine as B. abortus reservoirs in Starr County. Isolates from a 2005 cattle outbreak in Wyoming displayed VNTR-10 profiles matching those of strains recovered from Wyoming and Idaho elk. Additionally, isolates associated with cattle outbreaks in Idaho in 2002, Montana in 2008 and 2011, and Wyoming in 2010 primarily clustered with isolates recovered from GYA elk. This study indicates that elk play a predominant role in the transmission of B. abortus to cattle located in the GYA. PMID:22427502

  11. Serological and bacteriological responses of water buffalo (Bubalus bubalis) vaccinated with two doses of Brucella abortus strain RB51 vaccine.

    Science.gov (United States)

    Ramnanan, Anil; Diptee, Michael; Asgarali, Zinora; Campbell, Mervyn; Adesiyun, Abiodun Adewale

    2012-10-01

    Thirty-two water buffalo (Bubalus bubalis) calves aged 6–10 months were used to evaluate serological responses to Brucella abortus strain RB51 (RB51) vaccination in a dose-response study and to compare the use of two selective media for the isolation of RB51. The animals were randomly divided into three treatment groups. Groups I-III received the recommended vaccine dose (RD) twice 4 weeks apart, RD twice 18 weeks apart and saline once, respectively. Lymph nodes were excised from the three groups and subjected to bacteriological examination to determine the frequency of detection of RB51. Pre- and post-vaccination blood samples were collected and tested for B. abortus antibodies using the buffered plate agglutination test (BPAT), complement fixation test (CFT), and dot-blot assay. Sera taken at all post-inoculation weeks (PIW) were negative for field strain B. abortus using the BPAT. Antibody responses to RB51 were demonstrated in all vaccinates but not in controls by CFT and dot-blot assay from 1 PIW up to 16 weeks following booster vaccination. The agreement for both assays was 80.7% and there was a linear interdependence with a Pearson's correlation coefficient value of 0.578. The frequency of isolation of RB51 from the two selective media used was not significantly different (P > 0.05).

  12. 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.

  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

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    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. 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%.

  15. Diagnostic efficacy of Brucella abortus strain RB51 in experimentally inoculated Sprague-Dawley rats using western blot assay.

    Science.gov (United States)

    Rahman, Siddiqur; Baek, Byeong Kirl

    2008-10-01

    To investigate the diagnosis and efficacy of Brucella abortus strain RB51 (SRB51) in experimentally inoculated Sprague-Dawley (SD) rat using western blot assay. Female SD rats were orally administered with 1.0 x 10(7) colony forming unit (cfu) suspension of SRB51 and half of these SD rats were challenged at 4 weeks post inoculation with 1.0 x 10(9) cfu suspension of B. abortus biotype 1 isolated in South Korea. Sera of SD rats were monitored at regular intervals by western blot assay using whole cell antigen of B. abortus strain 1119-3 (S1119-3). The bacteriological examination of blood and clinical examination of the rats were also performed. There were several bands at 120, 70, 45, 30, 20 kDa and clear specific bands were found after vaccination (20, 70 kDa) and challenge (15, 20, 45, 70, 120 kDa). The highest immune response was observed in sera 4 weeks post SRB51 vaccination. SRB51 was recovered from the blood of all of SRB51 inoculated rats until one week post vaccination and there were no clinical signs in that inoculated rats. It is concluded that the SRB51 elicits antigen specific immunity in SD rats based on western blot assay.

  16. Overexpression of Protective Antigen as a Novel Approach To Enhance Vaccine Efficacy of Brucella abortus Strain RB51

    OpenAIRE

    Vemulapalli, Ramesh; He, Yongqun; Cravero, Silvio; Sriranganathan, Nammalwar; Boyle, Stephen M.; Schurig, Gerhardt G.

    2000-01-01

    Brucella abortus strain RB51 is an attenuated rough strain that is currently being used as the official live vaccine for bovine brucellosis in the United States and several other countries. We reasoned that overexpression of a protective antigen(s) of B. abortus in strain RB51 should enhance its vaccine efficacy. To test this hypothesis, we overexpressed Cu/Zn superoxide dismutase (SOD) protein of B. abortus in strain RB51. This was accomplished by transforming strain RB51 with a broad-host-r...

  17. Brucella abortus strain RB51 leucine auxotroph as an environmentally safe vaccine for plasmid maintenance and antigen overexpression.

    Science.gov (United States)

    Rajasekaran, Parthiban; Seleem, Mohamed N; Contreras, Andrea; Purwantini, Endang; Schurig, Gerhardt G; Sriranganathan, Nammalwar; Boyle, Stephen M

    2008-11-01

    To avoid potentiating the spread of an antibiotic resistance marker, a plasmid expressing a leuB gene and a heterologous antigen, green fluorescent protein (GFP), was shown to complement a leucine auxotroph of cattle vaccine strain Brucella abortus RB51, which protected CD1 mice from virulent B. abortus 2308 and elicited GFP antibodies.

  18. Evaluation and Selection of Multilocus Variable-Number Tandem-Repeat Analysis Primers for Genotyping Brucella abortus Biovar 1 Isolated from Human Patients.

    Science.gov (United States)

    Lee, Subok; Hwang, Kyu-Jam; Park, Mi-Yeoun; Hwang, Seon-Do; Chai, Hee-Youl; Chu, Hyuk; Park, Sang-Hee

    2013-10-01

    Brucellosis is the most common bacterial zoonosis in the world. Multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) is a molecular method for genotyping bacterial species. Brucella abortus biovar I was isolated from most of the brucellosis-suspected patients in Korea. This study was conducted to investigate the ability of various MLVA primers that are used for molecular typing B. abortus isolates and for analyzing their epidemiological data. A total of 80 human isolates of B. abortus biovar I isolated from human patients and the reference strain were used for MLVA. Genetic diversity was determined by calculating the Simpson's diversity index (DI) of each VNTR locus. The Brucella strains were subcultured 30 times to determine the stability of each locus. The DNA of the strains cultivated in each passage was extracted and subjected to MLVA for further investigation. The 15 VNTR loci were selected based on high DI values. The DIs of the 15 VNTR loci showed considerable discrimination power ranging from 59% for Bruce 43 to 87% for Bruce 22. Bruce 09, Bruce 11, Bruce 16, Bruce 42, and Bruce 43 were confirmed to remain stable in vitro among the 15 VNTR loci selected. The results of this study suggest that the five loci subsets may be a useful epidemiological tool for investigating B. abortus biovar 1 outbreak.

  19. Immune Responses and Protection against Experimental Brucella suis Biovar 1 Challenge in Nonvaccinated or B. abortus Strain RB51-Vaccinated Cattle▿

    OpenAIRE

    Olsen, S C; Hennager, S. G.

    2010-01-01

    Twenty Hereford heifers approximately 9 months of age were vaccinated with saline (control) or 2 × 1010 CFU of the Brucella abortus strain RB51 (RB51) vaccine. Immunologic responses after inoculation demonstrated significantly greater (P < 0.05) antibody and proliferative responses to RB51 antigens in cattle vaccinated with RB51 than in the controls. Pregnant cattle received a conjunctival challenge at approximately 6 months of gestation with 107 CFU of B. suis bv. 1 strains isolated from nat...

  20. Experimental Infection of Richardson's Ground Squirrels (Spermophilus richardsonii) with Attenuated and Virulent Strains of Brucella abortus

    Science.gov (United States)

    Exposure of non-target species to wildlife vaccines is an important concern when evaluating a candidate vaccine for use in the field. A previous investigation of the safety of Brucella abortus strain RB51 (sRB51) in various non-target species suggested that Richardson’s ground squirrels (Spermophil...

  1. 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...

  2. 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.

  3. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Characterization of Brucella abortus mutant strain Δ22915, a potential vaccine candidate.

    Science.gov (United States)

    Bao, Yanqing; Tian, Mingxing; Li, Peng; Liu, Jiameng; Ding, Chan; Yu, Shengqing

    2017-04-04

    Brucellosis, caused by Brucella spp., is an important zoonosis worldwide. Vaccination is an effective strategy for protection against Brucella infection in livestock in developing countries and in wildlife in developed countries. However, current vaccine strains including S19 and RB51 are pathogenic to humans and pregnant animals, limiting their use. In this study, we constructed the Brucella abortus (B. abortus) S2308 mutant strain Δ22915, in which the putative lytic transglycosylase gene BAB_RS22915 was deleted. The biological properties of mutant strain Δ22915 were characterized and protection of mice against virulent S2308 challenge was evaluated. The mutant strain Δ22915 showed reduced survival within RAW264.7 cells and survival in vivo in mice. In addition, the mutant strain Δ22915 failed to escape fusion with lysosomes within host cells, and caused no observable pathological damage. RNA-seq analysis indicated that four genes associated with amino acid/nucleotide transport and metabolism were significantly upregulated in mutant strain Δ22915. Furthermore, inoculation of ∆22915 at 10 5 colony forming units induced effective host immune responses and long-term protection of BALB/c mice. Therefore, mutant strain ∆22915 could be used as a novel vaccine candidate in the future to protect animals against B. abortus infection.

  5. Characterization of Recombinant B. abortus Strain RB51SOD Toward Understanding the Uncorrelated Innate and Adaptive Immune Responses Induced by RB51SOD Compared to Its Parent Vaccine Strain RB51

    OpenAIRE

    Zhu, Jianguo; Larson, Charles B.; Ramaker, Megan Ann; Quandt, Kimberly; Wendte, Jered M.; Ku, Kimberly P.; Chen, Fang; Jourdian, George W.; Vemulapalli, Ramesh; Schurig, Gerhardt G.; He, Yongqun

    2011-01-01

    Brucella abortus is a Gram-negative, facultative intracellular pathogen for several mammals, including humans. Live attenuated B. abortus strain RB51 is currently the official vaccine used against bovine brucellosis in the United States and several other countries. Overexpression of protective B. abortus antigen Cu/Zn superoxide dismutase (SOD) in a recombinant strain of RB51 (strain RB51SOD) significantly increases its vaccine efficacy against virulent B. abortus challenge in a mouse model. ...

  6. Characterization of recombinant B. abortus strain RB51SOD towards understanding the uncorrelated innate and adaptive immune responses induced by RB51SOD compared to its parent vaccine strain RB51

    OpenAIRE

    Jianguo eZhu; Jianguo eZhu; Charles Bradford Larson; Megan Ann Ramaker; Kimberly eQuandt; Jered eWendte; Kimberly eKu; Fang eChen; George eJourdian; Ramesh eVemulapalli; Gerhardt G. Schurig; Yongqun Oliver eHe

    2011-01-01

    Brucella abortus is a Gram-negative, facultative intracellular pathogen for several mammals, including humans. Live attenuated B. abortus strain RB51 is currently the official vaccine used against bovine brucellosis in the United States and several other countries. Overexpression of protective B. abortus antigen Cu/Zn superoxide dismutase (SOD) in a recombinant strain of RB51 (strain RB51SOD) significantly increases its vaccine efficacy against virulent B. abortus challenge in a mouse model. ...

  7. Shedding of Brucella abortus rough mutant strain RB51 in milk of water buffalo (Bubalus bubalis).

    Science.gov (United States)

    Longo, Mariangela; Mallardo, Karina; Montagnaro, Serena; De Martino, Luisa; Gallo, Sergio; Fusco, Giovanna; Galiero, Giorgio; Guarino, Achille; Pagnini, Ugo; Iovane, Giuseppe

    2009-07-01

    The objective of this study was to determine if Brucella abortus rough mutant strain RB51 (SRB51) is eliminated in buffalo milk. Thirty Brucella-free female buffaloes were used in this study: ten 4-5 years old were inoculated with the triple of the recommended calfhood dose of SRB51 by subcutaneous route, ten 2-3 years old at the first lactation were previously vaccinated twice as calves with triple the recommended calf dose of RB51, while five 4-5 years old and five 2-3 years old not vaccinated Brucella-free female buffaloes served as controls. Milk samples were taken aseptically on a daily basis for the first 30 days and weekly for the second and third months. The samples were inoculated on selective media for isolation of SRB51 and incubated for 11 days. Moreover, PCR analysis was also performed directly on milk samples. SRB51 was isolated from milk samples only during the first week post-vaccination while RB51 DNA was detected during the first week till the fourth week post-vaccination only in water buffaloes vaccinated as adults. The identification of Brucella RB51 in milk samples, strongly suggests that this Brucella vaccine could be excreted in milk of buffalo cows vaccinated as adults, while our data demonstrate that the vaccine is safe for use in buffaloes vaccinated as calves in which it was not excreted in milk.

  8. Efficacy of Dart or Booster Vaccination with Strain RB51 in Protecting Bison against Experimental Brucella abortus Challenge

    OpenAIRE

    Olsen, S. C.; Johnson, C. S.

    2012-01-01

    This study characterized the efficacy of the Brucella abortus strain RB51 vaccine in bison when delivered by single intramuscular vaccination (hand RB51), by single pneumatic dart delivery (dart RB51), or as two vaccinations approximately 13 months apart (booster RB51) in comparison to control bison. All bison were challenged intraconjunctivally in midgestation with 107 CFU of B. abortus strain 2308 (S2308). Bison were necropsied and sampled within 72 h of abortion or delivery of a live calf....

  9. Parenteral vaccination of domestic pigs with Brucella abortus strain RB51.

    Science.gov (United States)

    Stoffregen, William C; Olsen, Steven C; Bricker, Betsy J

    2006-10-01

    To determine the immunogenicity and efficacy of Brucella abortus strain RB51 (SRB51) as a vaccine in domestic pigs. Sixty-eight 6-week-old crossbred domestic pigs and twenty-four 4-month-old gilts. In experiment 1, pigs were vaccinated IM (n = 51) with 2 x 10(10) CFUs of SRB51 or sham inoculated (17). Periodic blood samples were obtained to perform blood cultures, serologic evaluations, and cell-mediated immunity assays. Necropsies were performed at selected times between weeks 1 and 23 after vaccination to determine vaccine clearance. In experiment 2, gilts were similarly vaccinated (n = 18) or sham inoculated (8) and similar samples were obtained after vaccination. Gilts were bred and challenged conjunctivally with 5.0 x 10(7) CFUs of virulent Brucella suis strain 3B. Necropsies were performed on gilts and on fetuses or neonates after abortion or parturition, respectively. Bacterial cultures and serologic evaluations were performed on samples obtained at necropsy to determine vaccine efficacy. Humoral and cell-mediated immune responses did not differ between vaccinates and controls. After vaccination, SRB51 was not isolated from blood cultures of either group and was isolated from lymphoid tissues of 3 pigs at 2 weeks (n = 2) and 4 weeks (1) after vaccination. No differences were found in isolation of B suis or in seroconversion between vaccinated and control gilts and between their neonates or aborted fetuses. Parenteral vaccination with SRB51 does not induce humoral or cell-mediated immune responses. Vaccination with SRB51 did not protect gilts or their neonates and fetuses from virulent challenge with B suis.

  10. Isolation of Chlamydia abortus from a laboratory worker diagnosed with atypical pneumonia.

    Science.gov (United States)

    Ortega, Nieves; Caro, M Rosa; Gallego, M Carmen; Murcia-Belmonte, Antonio; Álvarez, Daniel; Del Río, Laura; Cuello, Francisco; Buendía, Antonio J; Salinas, Jesús

    2015-01-01

    Identifying the aetiological agent of atypical pneumonia in human can sometimes be a tedious process, especially in cases where Mycoplasma pneumoniae, Legionella species and Chlamydia pneumoniae are ruled out. In such cases, a correct anamnesis of the patient is basic to clarify which pathogens might have produced the infection. For this reason, health professionals including veterinarians and laboratory personnel working with zoonotic pathogens should keep their doctors informed. A human case of atypical pneumonia linked to Chlamydia abortus is reported. A 47-year-old male, a veterinarian researcher into chlamydiae, developed respiratory symptoms, breathing problems and high fever. Serological analyses ruled out the involvement of several respiratory pathogens, such as M. pneumoniae, Legionella pneumophila, Rickettsia conorii and C. pneumoniae, and Chlamydia abortus was identified as the possible aetiological agent of the infection. The isolation of C. abortus from the patient's sputum and subsequent molecular analysis confirmed the presence of this microorganism. As far as we know, although C. abortus has not been previously described as capable of causing pneumonia in humans, this is the first reported case of atypical pneumonia in which C. abortus is thought to have played an aetiological role.

  11. Safety of revaccination of pregnant bison with Brucella abortus strain RB51.

    Science.gov (United States)

    Olsen, S C; Holland, S D

    2003-10-01

    From December 1998 through February 1999, a study was conducted in a Brucella-infected bison herd to evaluate the safety of booster vaccination of adult bison (Bison bison) with 6 x 10(9) colony forming units (CFU) of Brucella abortus strain RB51 (SRB51) that had previously been vaccinated as yearlings with 1 x 10(10) CFU of SRB51. Abortions or other adverse effects were not observed after SRB51 booster vaccination. At 10 wk after adult vaccination, pregnant and nonpregnant bison (n = 65) were randomly selected for bacteriologic sampling of targeted maternal tissues during abattoir processing. Fetal tissues were also sampled in pregnant bison. The SR351 recovered from tissue samples of eight of 48 pregnant bison and none of 17 nonpregnant bison. In three of the eight culture-positive bison, SRB51 was recovered from fetal tissues. In three additional bison, one pregnant and two nonpregnant, B. abortus biovar 1 field strain was recovered from internal iliac or supramammary lymphatic tissues. Results of this study suggest the possibility that the SRB51 vaccine can be safely used to booster vaccinate pregnant bison in a Brucella-infected bison herd. Our data also reaffirms the potential for B. abortus field strains to persist in bison until attainment of reproductive age, despite extensive use of vaccination and serologic testing.

  12. Brucella abortus strain RB51 vaccination in elk. I. Efficacy of reduced dosage.

    Science.gov (United States)

    Cook, Walter E; Williams, Elizabeth S; Thorne, E Tom; Kreeger, Terry J; Stout, Glen; Bardsley, Katie; Edwards, Hank; Schurig, Gerhardt; Colby, Lesley A; Enright, Fred; Elzer, Philip H

    2002-01-01

    Bovine brucellosis is a serious zoonotic disease affecting some populations of Rocky Mountain elk (Cervus elaphus nelsoni) and bison (Bison bison) in the Greater Yellowstone Area, USA. The fear that elk and/or bison may spread Brucella abortus to livestock has prompted efforts to reduce or eliminate the disease in wildlife. Brucella abortus strain RB51 (RB51) vaccine has recently been approved for use in cattle. Unlike strain 19 vaccine, RB51 does not cause false positive reactions on standard brucellosis serologic tests. If effective, it may become the vaccine of choice for wildlife. In February 1995, 45 serologically negative female elk calves were trapped and taken to the Sybille Wildlife Research and Conservation Education Unit near Wheatland, Wyoming, USA. In May 1995, 16 of these elk calves were hand-vaccinated with 1 x 10(9) colony forming units (CFU) of RB51, 16 were vaccinated with 1 x 10(8) CFU RB51 by biobullet, and 13 were given a saline placebo. The elk were bred in fall of 1996 and they were challenged with 1 x 10(7) CFU of B. abortus strain 2308 by intraconjunctival inoculation in March 1997. Thirteen (100%) control elk aborted, 14 (88%) hand-vaccinated elk aborted, and 12 (75%) biobullet vaccinated elk aborted or produced nonviable calves. These results suggest that a single dose of 1 x 10(8) to 1 x 10(9) CFU RB51 does not provide significant protection against B. abortus induced abortion in elk. However, the vaccine appears to be safe at this dose and additional study may reveal a more effective RB51 vaccine regimen for elk.

  13. Genome sequence of Brucella abortus vaccine strain S19 compared to virulent strains yields candidate virulence genes.

    Directory of Open Access Journals (Sweden)

    Oswald R Crasta

    Full Text Available The Brucella abortus strain S19, a spontaneously attenuated strain, has been used as a vaccine strain in vaccination of cattle against brucellosis for six decades. Despite many studies, the physiological and molecular mechanisms causing the attenuation are not known. We have applied pyrosequencing technology together with conventional sequencing to rapidly and comprehensively determine the complete genome sequence of the attenuated Brucella abortus vaccine strain S19. The main goal of this study is to identify candidate virulence genes by systematic comparative analysis of the attenuated strain with the published genome sequences of two virulent and closely related strains of B. abortus, 9-941 and 2308. The two S19 chromosomes are 2,122,487 and 1,161,449 bp in length. A total of 3062 genes were identified and annotated. Pairwise and reciprocal genome comparisons resulted in a total of 263 genes that were non-identical between the S19 genome and any of the two virulent strains. Amongst these, 45 genes were consistently different between the attenuated strain and the two virulent strains but were identical amongst the virulent strains, which included only two of the 236 genes that have been implicated as virulence factors in literature. The functional analyses of the differences have revealed a total of 24 genes that may be associated with the loss of virulence in S19. Of particular relevance are four genes with more than 60 bp consistent difference in S19 compared to both the virulent strains, which, in the virulent strains, encode an outer membrane protein and three proteins involved in erythritol uptake or metabolism.

  14. Vector Development for the Expression of Foreign Proteins in the Vaccine Strain Brucella abortus S19

    Science.gov (United States)

    Comerci, Diego J.; Pollevick, Guido D.; Vigliocco, Ana M.; Frasch, Alberto C. C.; Ugalde, Rodolfo A.

    1998-01-01

    A vector for the expression of foreign antigens in the vaccine strain Brucella abortus S19 was developed by using a DNA fragment containing the regulatory sequences and the signal peptide of the Brucella bcsp31 gene. This fragment was cloned in broad-host-range plasmid pBBR4MCS, resulting in plasmid pBEV. As a reporter protein, a repetitive antigen of Trypanosoma cruzi was used. The recombinant fusion protein is stably expressed and secreted into the Brucella periplasmic space, inducing a good antibody response against the T. cruzi antigen. The expression of the repetitive antigen in Brucella neither altered its growth pattern nor generated a toxic or lethal effect during experimental infection. The application of this strategy for the generation of live recombinant vaccines and the tagging of B. abortus S19 vaccine is discussed. This is the first time that a recombinant protein has been expressed in the periplasm of brucellae. PMID:9673273

  15. 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.

  16. Efficacy of strain RB51 vaccine in protecting infection and vertical transmission against Brucella abortus in Sprague-Dawley rats.

    Science.gov (United States)

    Islam, Md Ariful; Khatun, Mst Minara; Baek, Byeong-Kirl; Lee, Sung-Il

    2009-09-01

    Immunizing animals in the wild against Brucella (B.) abortus is essential to control bovine brucellosis because cattle can get the disease through close contact with infected wildlife. The aim of this experiment was to evaluate the effectiveness of the B. abortus strain RB51 vaccine in protecting infection as well as vertical transmission in Sprague-Dawley (SD) rats against B. abortus biotype 1. Virgin female SD rats (n = 48) two months of age were divided into two groups: one group (n = 24) received RB51 vaccine intraperitoneally with 3 x 10(10) colony forming units (CFU) and the other group (n = 24) was used as non-vaccinated control. Non-vaccinated and RB51-vaccinated rats were challenged with 1.5 x 10(9) CFU of virulent B. abortus biotype 1 six weeks after vaccination. Three weeks after challenge, all rats were bred. Verification of RB51-vaccine induced protection in SD rats was determined by bacteriological, serological and molecular screening of maternal and fetal tissues at necropsy. The RB51 vaccine elicited 81.25% protection in SD rats against infection with B. abortus biotype 1. Offspring from rats vaccinated with RB51 had a decreased (p RB51 vaccination efficacy against the vertical transmission of B. abortus in the SD rat model.

  17. 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 (PRB51). Similar results were observed on BALB/c groups. However, in Swiss mouse groups, S19 was more protective than RB51 (PRB51 vaccines. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Overexpression of Protective Antigen as a Novel Approach To Enhance Vaccine Efficacy of Brucella abortus Strain RB51

    Science.gov (United States)

    Vemulapalli, Ramesh; He, Yongqun; Cravero, Silvio; Sriranganathan, Nammalwar; Boyle, Stephen M.; Schurig, Gerhardt G.

    2000-01-01

    Brucella abortus strain RB51 is an attenuated rough strain that is currently being used as the official live vaccine for bovine brucellosis in the United States and several other countries. We reasoned that overexpression of a protective antigen(s) of B. abortus in strain RB51 should enhance its vaccine efficacy. To test this hypothesis, we overexpressed Cu/Zn superoxide dismutase (SOD) protein of B. abortus in strain RB51. This was accomplished by transforming strain RB51 with a broad-host-range plasmid, pBBR1MCS, containing the sodC gene along with its promoter. Strain RB51 overexpressing SOD (RB51SOD) was tested in BALB/c mice for its ability to protect against challenge infection with virulent strain 2308. Mice vaccinated with RB51SOD, but not RB51, developed antibodies and cell-mediated immune responses to Cu/Zn SOD. Strain RB51SOD vaccinated mice developed significantly (P RB51 alone. The presence of the plasmid alone in strain RB51 did not alter its vaccine efficacy. Also, overexpression of SOD did not alter the attenuation characteristic of strain RB51. PMID:10816475

  19. 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 ...

  20. 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...

  1. Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests.

    Science.gov (United States)

    Aguirre, N P; Vanzini, V R; Torioni de Echaide, S; Valentini, B S; De Lucca, G; Aufranc, C; Canal, A; Vigliocco, A; Nielsen, K

    2002-01-01

    The serological response induced by Brucella abortus strain 19 was evaluated in 52 Holstein females from a brucellosis-free herd using seven serological tests. Each calf was vaccinated at an age of 4 and 8 months old with 3 x 10(10) CFU B. abortus S19 and the antibody response was determined as the proportion of positive results to each test. The antibody dynamics, measured with the buffered plate antigen (BPA) test and the rapid automated presumptive (RAP) test, were similar. The proportion of positive reactions in these tests reached 100% one week after vaccination and remained at this level for seven weeks, after which the proportion of positive samples slowly declined to 8% (BPA) and 2% (RAP) at week 50. The response in the indirect enzyme immunoassay (i-ELISA) was similar, but shorter than that observed with the BPA/RAP. The antibody dynamic, measured using the seroagglutination test (SAT) in parallel with the 2-mercaptoethanol (2-Me) test and the complement fixation test (CFT) were similar to the RAP/BPA, but of slightly shorter duration. The competitive ELISA (c-ELISA) was positive in all animals for 3 weeks, followed by a rapid decline. The fluorescence polarization assay (FPA) reached a maximum of 68.5% positive animals at week 4 and then declined. Based on these data, the c-ELISA and FPA discriminated residual antibody activity due to vaccination more efficiently than the other tests.

  2. Brucella abortus vaccine strain RB51 produces low levels of M-like O-antigen.

    Science.gov (United States)

    Cloeckaert, Axel; Zygmunt, Michel S; Guilloteau, Laurence A

    2002-03-15

    Brucella abortus RB51 is a rough (R) stable vaccine strain used in cattle and is believed to be devoid of O-side chain. We analyzed by use of a panel of monoclonal antibodies (MAbs) directed against seven previously defined O-polysaccharide (O-PS) epitopes the O-chain expression in strain RB51. Two MAbs specific for the C/Y (A=M) and C (M>A) epitopes showed low bindings in ELISA to strain RB51. O-chain expression was further confirmed by Western blot after SDS-PAGE of strain RB51. In particular, the MAb of C (M>A) specificity, showing preferential binding to M-dominant smooth (S) Brucella strains, revealed in strain RB51 a typical smooth-lipopolysaccharide (S-LPS) pattern which resembled that of M-dominant S-LPS. Thus, the results clearly show that strain RB51 produces low levels of M-like O-antigen.

  3. Experimental infection of nontarget species of rodents and birds with Brucella abortus strain RB51 vaccine

    Science.gov (United States)

    Januszewski, M.C.; Olsen, S.C.; McLean, R.G.; Clark, L.; Rhyan, Jack C.

    2001-01-01

    The Brucella abortus vaccine strain RB51 (SRB51) is being considered for use in the management of bnucellosis in wild bison (Bison bison) and elk (Cervus elaphus) populations in the Greater Yellowstone Area (USA). Evaluation of the vaccines safety in non-target species was considered necessary prior to field use. Between June 1998 and December 1999, ground squirrels (Spermophilus richardsonii, n = 21), deer mice (Peromyscus maniculatus, n = 14), prairie voles (Microtus ochrogaster, n = 21), and ravens (Corvus corax, n = 13) were orally inoculated with SRB51 or physiologic saline. Oral and rectal swabs and blood samples were collected for bacteriologic evaluation. Rodents were necropsied at 8 to 10 wk and 12 to 21 wk post inoculation (PI), and ravens at 7 and 11 wk PI. Spleen, liver and reproductive tissues were collected for bacteriologic and histopathologic evaluation. No differences in clinical signs, appetite, weight loss or gain, or activity were observed between saline- and SRB51-inoculated animals in all four species. Oral and rectal swabs from all species were negative throughout the study. In tissues obtained from SRB51-inoculated animals, the organism was isolated from six of seven (86%) ground squirrels, one of six (17%) deer mice, none of seven voles, and one of five (20%) ravens necropsied at 8, 8, 10, and 7 wk PI, respectively. Tissues from four of seven (57%) SRB51-inoculated ground squirrels were culture positive for the organism 12 wk PI; SRB51 was not recovered from deer mice, voles. or ravens necropsied 12, 21, or 11 wk, respectively, PI. SRB51 was not recovered from saline-inoculated ground squirrels, deer mice, or voles at any time but was recovered from one saline-inoculated raven at necropsy, 7 wk PI, likely attributable to contact with SRB51-inoculated ravens in an adjacent aviary room. Spleen was time primary tissue site of colonization in ground squirrels, followed by the liver and reproductive organs. The results indicate oral exposure to

  4. The Humoral Immune Response of Elks (Cervus elaphus nelsoni) and Mice to Vaccination with Brucella abortus Strain RB51

    OpenAIRE

    Colby, Lesley A.

    1997-01-01

    Vaccine Brucella abortus strain RB51, unlike the wild strain 2308 and another vaccine strain (strain 19) does not induce anti-O-chain antibodies. An efficacious vaccine strain that fails to produce an O-chain and thus a lack of an anti-O-chain humoral response greatly simplifies identification of vaccinated versus field strain infected animals. The three primary objectives of this research were the following: 1) to develop a serological assay to detect anti-RB51 antibodies in vaccinated elk (...

  5. Isolation of Brucella abortus Using PCR-RFLP Analysis

    OpenAIRE

    M Salehi; E Pishva; R Salehi; R Rahmani

    2006-01-01

    Brucella transmission and epidemiology depend on infecting species and biovar. Therefore, exact identification of the Brucella is important to design correct control and treatment strategies. In this study, we examined presence of other Brucellae in Isfahan. One hundred twenty Brucella isolates were collected and genomic DNA was extracted from them. omp2a fragment of all isolates were amplified using a pair of specific primers and the PCR products were electrophoresed and stained with EtBr. T...

  6. 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 RB51-vaccinated bison at 8, 16, and 24 weeks after the initial vaccination and at 8 weeks after the booster vaccination. The vaccinated bison had 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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  7. Protection to respiratory challenge of Brucella abortus strain 2308 in the lung.

    Science.gov (United States)

    Surendran, Naveen; Sriranganathan, Nammalwar; Boyle, Stephen M; Hiltbold, Elizabeth M; Tenpenny, Nancy; Walker, Michelle; Zimmerman, Kurt; Werre, Stephen; Witonsky, Sharon G

    2013-08-28

    Brucella is amongst the top 5 causes of zoonotic disease worldwide. Infection is through ingestion, inhalation or contact exposure. Brucella is characterized as a class B pathogen by Centers of Disease Control and Prevention (CDC). Currently, there are no efficacious vaccines available in people. Currently available USDA approved vaccines for animals include B. abortus strain RB51 and B. melitensis Rev1. Protection is mediated by a strong innate and CD4 Th1, CD8 Tc1 immune response. If protective vaccines can be developed, disease in people and animals can be controlled. While strain RB51 protects in cattle, and against intraperitoneal challenge in mice, it does not protect against respiratory challenge. Therefore, we assessed the efficacy of strain RB51 combined with different TLR agonists, and O-side chain from LPS, to enhance protection against respiratory challenge with strain 2308. We hypothesized that TLR agonists and O-side chain would enhance protection. Strains RB51 with TLR2 agonist, RB51 with TLR4 agonist and strain 19 provided significant protection in the lung. Protection using strain RB51 with TLR agonists was associated with increased IgG2a and IgG1 in the (bronchoalveolar lavage) BAL and serum, and increased IgA (serum). Splenocytes from strain RB51 with TLR2 vaccinated mice up-regulated antigen specific interferon-gamma and TNF-alpha production. Vaccination and challenge resulted in significant increases in activated dendritic cells (DCs), and increased CD4 and CD8 cells in the BAL. Overall, this study demonstrates the ability of TLR agonists 2 and 4 to up-regulate strain RB51 mediated protection in the lung to respiratory challenge against strain 2308. Copyright © 2013. Published by Elsevier Ltd.

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Science.gov (United States)

    Adone, Rosanna; Muscillo, Michele; La Rosa, Giuseppina; Francia, Massimiliano; Tarantino, Michela

    2011-01-01

    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.

  11. Tipificación molecular de Brucella abortus cepa 19 y su aplicación al control de biológicos Molecular characterization of Brucella abortus strain 19 and its application for controlling biologics

    Directory of Open Access Journals (Sweden)

    M.E. Pavan

    2005-09-01

    Full Text Available Brucella abortus es el agente etiológico de la brucelosis bovina. La cepa 19, utilizada en la elaboración de vacunas, puede ser identificada a través de una deleción en la región eri asociada con la sensibilidad al eritritol. Se optimizó un ensayo de PCR para caracterizar específicamente esta cepa. El método que describimos es un procedimiento rápido para identificar B. abortus y simultáneamente diferenciar la cepa 19 de otras cepas de B. abortus biovar 1. Hemos aplicado este ensayo para la detección de la cepa 19 en vacunas contra la brucelosis bovina elaboradas en Argentina. Los resultados indican que este método podría ser útil para el seguimiento de las cepas madres y semillas utilizadas en la producción industrial de esta vacuna. Esta metodología también contribuiría a la reducción del riesgo de la infección adquirida en el laboratorio y podría aplicarse como prueba de rutina para confirmar la presencia de B. abortus en vacunas no relacionadas.Brucella abortus is the etiological agent of bovine brucellosis. The strain 19 used in vaccine elaboration can be identified through a deletion in the eri region associated with its susceptibility to erythritol. We optimized a PCR assay for specific characterization of this strain. The method described here is a rapid procedure that enables identification of B. abortus, and simultaneous differentiation of the strain 19 from other B. abortus biovar 1 strains. We applied the assay to detect the strain 19 in vaccines against B. abortus produced in Argentina. The results show this method could be used to follow vaccine seed cultures of this strain. The methodology could also contribute to reduce the risk of a laboratory-acquired infection and could be of great help as a routine test for confirmation of B. abortus in non related vaccines.

  12. Comparison of Abortion and Infection after Experimental Challenge of Pregnant Bison and Cattle with Brucella abortus Strain 2308▿

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    Olsen, S. C.; Johnson, C.

    2011-01-01

    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 during midgestation. The incidence of abortion, fetal infection, uterine or mammary infection, or infection in maternal tissues after experimental challenge was greater (P abort, the time between experimental challenge and abortion was shorter (P abortion did not differ (P > 0.05) between cattle and bison. The results of our study suggest that naive bison and cattle have similarities and differences after experimental exposure to a virulent B. abortus strain. Although our data suggest that bison may be more susceptible to infection with Brucella, some pathogenic characteristics of brucellosis were similar between bison and cattle. PMID:21976222

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

    Science.gov (United States)

    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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Effectiveness of Brucella abortus Strain 19 single calfhood vaccination in elk (Cervus elaphus)

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    Roffe, Thomas J.; Jones, Lee C.; Coffin, Kenneth; Sweeney, Steven J.; Williams, Beth; Quist, Charlotte

    2002-01-01

    Brucellosis in Greater Yellowstone Area (GYA) bison and elk has been a source of controversy and focus of the Greater Yellowstone Interagency Brucellosis Committee (GYIBC) for years. Brucellosis has been eradicated from cattle in the 3 states of Wyoming, Montana, and Idaho and all three states currently are classified as “brucellosis free” with regard to livestock. Yet free-ranging elk that attend feedgrounds in the GYA, and bison in Yellowstone and Grand Teton National Parks, still have high seroprevalence to the disease and are viewed as a threat to the state-federal cooperative national brucellosis eradication program. Recently, cattle in eastern Idaho were found infected with brucellosis and transmission was apparently from fed elk. The GYIBC, formed of state and federal agencies involved in wildlife and livestock management in the 3 states, has committed to eventual elimination of the disease from wildlife. Management tools to control or eliminate the disease are limited; however, wildlife vaccination is one of the methods currently employed. Effective wildlife vaccination depends on dose efficacy, deliverability, and safety to non-targeted species. We commenced a single-dose efficacy study of vaccine Brucella abortus strain 19 (S19) in elk in 1999.

  15. Safety of Brucella abortus strain RB51 vaccine in non-target ungulates and coyotes.

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    Kreeger, Terry J; DeLiberto, Thomas J; Olsen, Steven C; Edwards, William H; Cook, Walter E

    2002-07-01

    Brucellosis is endemic in free-ranging elk (Cervus elaphus) and bison (Bison bison) in the Greater Yellowstone Area (GYA; USA). It is possible that an oral brucellosis vaccine could be developed and disseminated in the GYA to reduce disease transmission. Should this occur, non-target species other than elk and bison may come in contact with the vaccine resulting in morbidity or mortality. To assess biosafety, bighorn sheep (Ovis canadensis; n = 10), pronghorn (Antilocapra americana; n = 9), mule deer (Odocoileus hemionus; n = 11), moose (Alces alces shirasi; n = 10), and coyotes (Canis latrans; n = 24) were given a single oral dose of at least 1.0 x 10(10) colony-forming units of Brucella abortus strain RB51 vaccine (RB51). Animals were randomly divided into vaccinated and control groups. Ungulates were captured, blood sampled, and swabs taken from the nares, rectum, and vagina for bacterial culture on day 0, 42, and 84 post-inoculation (PI). On day 42, the vaccinated group became a control group and vice versa in a crossover design. Blood and swab samples were taken from coyotes on days 0, 14, 28, and 42 PI. There was no crossover for the coyote study. Two coyotes from each group were also euthanized and cultured for RB51 on days 42, 84, 168, and 336 PI. Blood samples were analyzed for hematologic changes and antibodies to RB51 using a modified dot-blot assay. No morbidity or mortality as a result of vaccination was observed in any animal. There were no differences in hematologic parameters at any time for ungulate species; vaccinated coyotes had higher hematocrit, hemoglobin, and eosinophil counts (P RB51. Strain RB51 was cultured from oropharyngeal lymph nodes from one coyote 42 days PI and from a moose 117 days PI. This study suggested that a single oral dose of RB51 was safe in these species.

  16. Identification of an IS711 Element Interrupting the wboA Gene of Brucella abortus Vaccine Strain RB51 and a PCR Assay To Distinguish Strain RB51 from Other Brucella Species and Strains

    Science.gov (United States)

    Vemulapalli, Ramesh; McQuiston, John R.; Schurig, Gerhardt G.; Sriranganathan, Nammalwar; Halling, Shirley M.; Boyle, Stephen M.

    1999-01-01

    Brucella abortus vaccine strain RB51 is a natural stable attenuated rough mutant derived from the virulent strain 2308. The genetic mutations that are responsible for the roughness and the attenuation of strain RB51 have not been identified until now. Also, except for an assay based on pulsed-field gel electrophoresis, no other simple method to differentiate strain RB51 from its parent strain 2308 is available. In the present study, we demonstrate that the wboA gene encoding a glycosyltransferase, an enzyme essential for the synthesis of O antigen, is disrupted by an IS711 element in B. abortus vaccine strain RB51. Exploiting this feature, we developed a PCR assay that distinguishes strain RB51 from all other Brucella species and strains tested. PMID:10473532

  17. Safety and immunogenicity of Brucella abortus strain RB51 vaccine in cross bred cattle calves in India.

    Science.gov (United States)

    Singh, Rashmi; Basera, Sanjay Singh; Tewari, Kamal; Yadav, Shweta; Joshi, Sumit; Singh, Brajesh; Mukherjee, Falguni

    2012-03-01

    Safety and immunogenicity of Brucella abortus RB51 vaccine has been evaluated in an organised dairy farm in India. All the cattle (r = 29) vaccinated with strain RB51 'responded' to the vaccine as demonstrated by iELISA using acetone killed strain RB51 antigen. The percentage responders at day 35, 60 and 90 post vaccination were 100%, 95% and 20%, respectively. Strain RB51 was able to elicit a good IFN-gamma response from vaccinated animals. The post-vaccination time point analysis indicated that the cumulative IFN-gamma response of whole blood from vaccinates stimulated with heat killed RB51 antigen was elicited in 80% of calves at 60 days post vaccination. Absence of strain RB51 in the secretions and excretion and lack of local or systemic reaction indicated the safety of the vaccine.

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

    Science.gov (United States)

    Olsen, S C; Johnson, C S

    2012-06-01

    This study characterized the efficacy of the Brucella abortus strain RB51 vaccine in bison when delivered by single intramuscular vaccination (hand RB51), by single pneumatic dart delivery (dart RB51), or as two vaccinations approximately 13 months apart (booster RB51) in comparison to control bison. All bison were challenged intraconjunctivally in midgestation with 10(7) CFU of B. abortus strain 2308 (S2308). Bison were necropsied and sampled within 72 h of abortion or delivery of a live calf. Compared to nonvaccinated bison, bison in the booster RB51 treatment had a reduced (P RB51 and dart RB51) did not differ (P > 0.05) from the control group in the incidence of abortion or recovery of S2308 from uterine, mammary, fetal, or maternal tissues at necropsy. Compared to nonvaccinated animals, all RB51 vaccination groups had reduced (P RB51 group having reduced (P RB51 enhances protective immunity against Brucella challenge compared to single vaccination with RB51 by hand or by pneumatic dart. Our study also suggests that an initial vaccination of calves followed by booster vaccination as yearlings should be an effective strategy for brucellosis control in bison.

  19. Serological response to administration of Brucella abortus strain RB51 vaccine in beef and dairy heifers, using needle-free and standard needle-based injection systems

    Science.gov (United States)

    The purpose of this study was to compare immunologic responses of heifers vaccinated with 10**10 colony-forming units (CFU) of Brucella abortus strain RB51 (SRB51) by standard needle-and-syringe system or a needle-free injection system. Heifers were randomly assigned to control and vaccination gro...

  20. 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

  1. 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...

  2. Origins and global context of Brucella abortus in Italy.

    Science.gov (United States)

    Garofolo, Giuliano; Di Giannatale, Elisabetta; Platone, Ilenia; Zilli, Katiuscia; Sacchini, Lorena; Abass, Anna; Ancora, Massimo; Cammà, Cesare; Di Donato, Guido; De Massis, Fabrizio; Calistri, Paolo; Drees, Kevin P; Foster, Jeffrey T

    2017-02-02

    Brucellosis is a common and chronic disease of cattle and other bovids that often causes reproductive disorders. Natural infection in cattle is caused by Brucella abortus and transmission typically occurs during abortions, calving, or nursing. Brucellosis is also a major zoonotic disease due to contamination of dairy products or contact with the tissues of infected animals. Brucellosis has been eradicated from most of the developed world in the last 40 years but persists in many regions-the disease remains prevalent in portions of Africa, the Middle East, Asia, and Central and South America, as well as in the Mediterranean basin. In Italy, B. abortus has persisted in southern regions in both cattle and water buffalo. Previous attempts at analyzing the phylogenetics of B. abortus in Italy have been challenging due to limited genetic variability and unresolved global population genetic structure of this pathogen. We conducted genome-wide phylogenetic analyses on 11 representative strains of B. abortus from Italy, and compared these sequences to a worldwide collection of publically available genomes. Italian isolates belong to three clades that are basal to the main and global B. abortus lineage. Using six SNP-based assays designed to identify substructure within the Italian clades, we surveyed a collection of 261 isolates and found that one clade predominates throughout endemic districts in the country, while the other two clades are more geographically restricted to portions of southern Italy. Although related strains exist worldwide, B. abortus isolates from Italy are substantially different than those found in much of the rest of Europe and North America, and are more closely related to strains from the Middle East and Asia. Our assays targeting genetic substructure within Italy allowed us to identify the major lineages quickly and inexpensively, without having to generate whole genome sequences for a large isolate collection. These findings highlight the

  3. Immune responses and protection against experimental Brucella suis biovar 1 challenge in nonvaccinated or B. abortus strain RB51-vaccinated cattle.

    Science.gov (United States)

    Olsen, S C; Hennager, S G

    2010-12-01

    Twenty Hereford heifers approximately 9 months of age were vaccinated with saline (control) or 2 × 10(10) CFU of the Brucella abortus strain RB51 (RB51) vaccine. Immunologic responses after inoculation demonstrated significantly greater (P RB51 antigens in cattle vaccinated with RB51 than in the controls. Pregnant cattle received a conjunctival challenge at approximately 6 months of gestation with 10(7) CFU of B. suis bv. 1 strains isolated from naturally infected cattle. The fluorescence polarization assay and the buffered acid plate agglutination test had the highest sensitivities in detecting B. suis-infected cattle between 2 and 12 weeks after experimental infection. Serologic responses and lymphocyte proliferative responses to B. suis antigens did not differ between control and RB51 vaccinees after experimental infection. No abortions occurred in cattle in either treatment group after challenge, although there appeared to be an increased incidence of retained placenta after parturition in both the control and the RB51 vaccination treatment groups. Our data suggest that the mammary gland is a preferred site for B. suis localization in cattle. Vaccination with RB51 did not reduce B. suis infection rates in maternal or fetal tissues. In conclusion, although B. suis is unlikely to cause abortions and fetal losses in cattle, our data suggest that RB51 vaccination will not protect cattle against B. suis infection after exposure.

  4. Immune responses and safety after dart or booster vaccination of bison with Brucella abortus strain RB51.

    Science.gov (United States)

    Olsen, S C; Johnson, C

    2012-05-01

    One alternative for management of brucellosis in Yellowstone National Park bison (Bison bison) is vaccination of calves and yearlings. Although Brucella abortus strain RB51 vaccination protects bison against experimental challenge, the effect of booster vaccinations was unknown. This study characterized immunologic responses after dart or booster vaccination of bison with Brucella abortus strain RB51. In two studies, 8- to 10-month-old female bison were inoculated with saline (n = 14), hand vaccinated with 1.1 × 10(10) to 2.0 × 10(10) CFU of RB51 (n = 21), or dart vaccinated with 1.8 × 10(10) CFU of RB51 (n = 7). A subgroup of hand vaccinates in study 1 was randomly selected for booster vaccination 15 months later with 2.2 × 10(10) CFU of RB51. Compared to single vaccinates, booster-vaccinated bison had greater serologic responses to RB51. However, there was a trend for antigen-specific proliferative responses of peripheral blood mononuclear cells (PBMC) from booster vaccinates to be reduced compared to responses of PBMC from single vaccinates. PBMC from booster vaccinates tended to have greater gamma interferon (IFN-γ) production than those from single vaccinates. In general, dart vaccination with RB51 induced immunologic responses similar to those of hand vaccination. All vaccinates (single hand, dart, or booster) demonstrated greater (P RB51 in early gestation did not induce abortion or fetal infection. Our data suggest that booster vaccination does not induce strong anamnestic responses. However, phenotypic data on resistance to experimental challenge are required to fully assess the effect of booster vaccination on protective immunity.

  5. 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.

  6. Live Brucella abortus rough vaccine strain RB51 stimulates enhanced innate immune response in vitro compared to rough vaccine strain RB51SOD and virulent smooth strain 2308 in murine bone marrow-derived dendritic cells.

    Science.gov (United States)

    Surendran, Naveen; Hiltbold, Elizabeth M; Heid, Bettina; Sriranganathan, Nammalwar; Boyle, Stephen M; Zimmerman, Kurt L; Makris, Melissa R; Witonsky, Sharon G

    2011-01-10

    Brucella spp. are Gram-negative, coccobacillary, facultative intracellular pathogens. B. abortus strain 2308 is a pathogenic strain affecting cattle and humans. Rough B. abortus strain RB51, which lacks the O-side chain of lipopolysaccharide (LPS), is the live attenuated USDA approved vaccine for cattle in the United States. Strain RB51SOD, which overexpresses Cu-Zn superoxide dismutase (SOD), has been shown to confer better protection than strain RB51 in a murine model. Protection against brucellosis is mediated by a strong CD4+ Th(1) and CD8+ Tc(1) adaptive immune response. In order to stimulate a robust adaptive response, a solid innate immune response, including that mediated by dendritic cells, is essential. As dendritic cells (DCs) are highly susceptible to Brucella infection, it is possible that pathogenic strains could limit the innate and thereby adaptive immune response. By contrast, vaccine strains could limit or bolster the innate and subsequent adaptive immune response. Identifying how Brucella vaccines stimulate innate and adaptive immunity is critical for enhancing vaccine efficacy. The ability of rough vaccine strains RB51 and RB51SOD to stimulate DC function has not been characterized. We report that live rough vaccine strain RB51 induced significantly better (p ≤ 0.05) DC maturation and function compared to either strain RB51SOD or smooth virulent strain 2308, based on costimulatory marker expression and cytokine production. Copyright © 2010. Published by Elsevier B.V.

  7. Characterization of recombinant B. abortus strain RB51SOD towards understanding the uncorrelated innate and adaptive immune responses induced by RB51SOD compared to its parent vaccine strain RB51

    Directory of Open Access Journals (Sweden)

    Jianguo eZhu

    2011-11-01

    Full Text Available Brucella abortus is a Gram-negative, facultative intracellular pathogen for several mammals, including humans. Live attenuated B. abortus strain RB51 is currently the official vaccine used against bovine brucellosis in the United States and several other countries. Overexpression of protective B. abortus antigen Cu/Zn superoxide dismutase (SOD in a recombinant strain of RB51 (strain RB51SOD significantly increases its vaccine efficacy against virulent B. abortus challenge in a mouse model. An attempt has been made to better understand the mechanism of the enhanced protective immunity of RB51SOD compared to its parent strain RB51. We previously reported that RB51SOD stimulated enhanced Th1 immune response. In this study, we further found that T effector cells derived from RB51SOD-immunized mice exhibited significantly higher cytotoxic T lymphocyte (CTL activity than T effector cells derived from RB51-immunized mice against virulent B. abortus-infected target cells. Meanwhile, the macrophage responses to these two strains were also studied. Compared to RB51, RB51SOD cells had a lower survival rate in macrophages and induced lower levels of macrophage apoptosis and necrosis. The decreased survival of RB51SOD cells correlates with the higher sensitivity of RB51SOD, compared to RB51, to the bactericidal action of either Polymyxin B or sodium dodecyl sulfate (SDS. Furthermore, a physical damage to the outer membrane of RB51SOD was observed by electron microscopy. Possibly due to the physical damage, overexpressed Cu/Zn SOD in RB51SOD was found to be released into the bacterial cell culture medium. Therefore, the stronger adaptive immunity induced by RB51SOD did not correlate with the low level of innate immunity induced by RB51SOD compared to RB51. This unique and apparently contradictory profile is likely associated with the differences in outer membrane integrity and Cu/Zn SOD release.

  8. Characterization of recombinant B. abortus strain RB51SOD toward understanding the uncorrelated innate and adaptive immune responses induced by RB51SOD compared to its parent vaccine strain RB51.

    Science.gov (United States)

    Zhu, Jianguo; Larson, Charles B; Ramaker, Megan Ann; Quandt, Kimberly; Wendte, Jered M; Ku, Kimberly P; Chen, Fang; Jourdian, George W; Vemulapalli, Ramesh; Schurig, Gerhardt G; He, Yongqun

    2011-01-01

    Brucella abortus is a Gram-negative, facultative intracellular pathogen for several mammals, including humans. Live attenuated B. abortus strain RB51 is currently the official vaccine used against bovine brucellosis in the United States and several other countries. Overexpression of protective B. abortus antigen Cu/Zn superoxide dismutase (SOD) in a recombinant strain of RB51 (strain RB51SOD) significantly increases its vaccine efficacy against virulent B. abortus challenge in a mouse model. An attempt has been made to better understand the mechanism of the enhanced protective immunity of RB51SOD compared to its parent strain RB51. We previously reported that RB51SOD stimulated enhanced Th1 immune response. In this study, we further found that T effector cells derived from RB51SOD-immunized mice exhibited significantly higher cytotoxic T lymphocyte activity than T effector cells derived from RB51-immunized mice against virulent B. abortus-infected target cells. Meanwhile, the macrophage responses to these two strains were also studied. Compared to RB51, RB51SOD cells had a lower survival rate in macrophages and induced lower levels of macrophage apoptosis and necrosis. The decreased survival of RB51SOD cells correlates with the higher sensitivity of RB51SOD, compared to RB51, to the bactericidal action of either Polymyxin B or sodium dodecyl sulfate (SDS). Furthermore, a physical damage to the outer membrane of RB51SOD was observed by electron microscopy. Possibly due to the physical damage, overexpressed Cu/Zn SOD in RB51SOD was found to be released into the bacterial cell culture medium. Therefore, the stronger adaptive immunity induced by RB51SOD did not correlate with the low level of innate immunity induced by RB51SOD compared to RB51. This unique and apparently contradictory profile is likely associated with the differences in outer membrane integrity and Cu/Zn SOD release.

  9. Brucella abortus strain RB51 vaccine: immune response after calfhood vaccination and field investigation in Italian cattle population.

    Science.gov (United States)

    Tittarelli, Manuela; Bonfini, Barbara; De Massis, Fabrizio; Giovannini, Armando; Di Ventura, Mauro; Nannini, Donatella; Caporale, Vincenzo

    2008-01-01

    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.

  10. An ELISA for the evaluation of gamma interferon production in cattle vaccinated with Brucella abortus strain RB51.

    Science.gov (United States)

    Tittarelli, Manuela; De Massis, Fabrizio; Bonfini, Barbara; Di Ventura, Mauro; Scacchia, Massimo

    2009-01-01

    The results of an enzyme-linked immunosorbent assay (ELISA) implemented for the detection of gamma interferon (gamma interferon) production in cattle vaccinated with Brucella abortus strain RB51 are presented. A purified protein fraction derived from RB51 (RB51 brucellin) has been used as antigenic stimulus for whole blood. The test was evaluated for 300 days in ten heifers vaccinated at calfhood with 10 x 10(9) colony-forming units of RB51 and in five control heifers. All animals came from officially brucellosis-free herds. Vaccinated animals started to give positive results from day 17 post vaccination (pv) until day 239 pv. All vaccinated animals gave a positive reaction at least once (with a stimulation index exceeding 2.5). Nevertheless, if sampling on day 20 pv is excluded (90% of vaccinated animals gave positive results), the sensitivity of the test varies from 20% to 70%, with a 40% average. A stimulation index over 2.5 was also recorded in three control animals. The results suggest that the gamma-interferon test is not suitable for the detection of cattle vaccinated with RB51, either at the individual or at the herd level.

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

    Directory of Open Access Journals (Sweden)

    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.

  12. An ELISA for the evaluation of gamma interferon production in cattle vaccinated with Brucella abortus strain RB51

    Directory of Open Access Journals (Sweden)

    Manuela Tittarelli

    2009-06-01

    Full Text Available The results of an enzyme-linked immunosorbent assay (ELISA implemented for the detection of gamma interferon (g-interferon production in cattle vaccinated with Brucella abortus strain RB51 are presented. A purified protein fraction derived from RB51 (RB51 brucellin has been used as antigenic stimulus for whole blood. The test was evaluated for 300 days in ten heifers vaccinated at calfhood with 10 × 109 colony-forming units of RB51 and in five control heifers. All animals came from officially brucellosis-free herds. Vaccinated animals started to give positive results from day 17 post vaccination (pv until day 239 pv. All vaccinated animals gave a positive reaction at least once (with a stimulation index exceeding 2.5. Nevertheless, if sampling on day 20 pv is excluded (90% of vaccinated animals gave positive results, the sensitivity of the test varies from 20% to 70%, with a 40% average. A stimulation index over 2.5 was also recorded in three control animals. The results suggest that the g-interferon test is not suitable for the detection of cattle vaccinated with RB51, either at the individual or at the herd level.

  13. Immune Responses and Protection against Experimental Challenge after Vaccination of Bison with Brucella abortus Strain RB51 or RB51 Overexpressing Superoxide Dismutase and Glycosyltransferase Genes▿

    OpenAIRE

    Olsen, S. C.; Boyle, S. M.; Schurig, G. G.; Sriranganathan, N. N.

    2009-01-01

    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 strain RB51 (RB51) or a recombinant RB51 strain overexpressing superoxide dismutase (sodC) and glycosyltransferase (wboA) genes (RB51+sodC,wboA). Bison were vaccinated with saline only or with 4.6 × 101...

  14. Lymphocyte proliferation in response to Brucella abortus 2308 or RB51 antigens in mice infected with strain 2308, RB51, or 19.

    OpenAIRE

    Stevens, M G; Olsen, S C; Pugh, G W

    1994-01-01

    Lymphocyte proliferation to 22 protein fractions (106 to 18 kDa) of Brucella abortus 2308 or the lipopolysaccharide O-antigen-deficient mutant of 2308, strain RB51, was measured for 20 weeks after infection of mice with strain 2308, RB51, or 19. Throughout the 20-week study, the 22 protein fractions of 2308 and RB51 induced a similar pattern of proliferation when they were incubated with lymphocytes from the infected mice. In addition, during the 20 weeks, lymphocytes from all groups of infec...

  15. Genetic stability of Brucella abortus S19 and RB51 vaccine strains by multiple locus variable number tandem repeat analysis (MLVA16).

    Science.gov (United States)

    Dorneles, Elaine Maria Seles; de Faria, Ana Paula Paiva; Pauletti, Rebeca Barbosa; Santana, Jordana Almeida; Caldeira, George Afonso Vítor; Heinemann, Marcos Bryan; Titze-de-Almeida, Ricardo; Lage, Andrey Pereira

    2013-10-01

    The aims of the present study were (i) to assess the in vitro genetic stability of S19 and RB51 Brucella abortus vaccines strains and (ii) to evaluate the ability of multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA) as a tool to be used in the quality control of live vaccines against brucellosis. Sixty-three batches of commercial S19 (n=53) and RB51 (n=10) vaccines, produced between 2006 and 2009, were used in this study. S19 and RB51 vaccines were obtained from, respectively, seven and two different manufacturers. Ten in vitro serial passages were performed on reference strains and on selected batches of commercial vaccines. All batches, reference strains and strains of serial passages were typed by the MLVA16. The results demonstrated that B. abortus S19 and RB51 vaccine strains are genetically stable and very homogeneous in their respective groups. Anyway, batches of S19 from one manufacturer and batches of RB51 from another presented genotypes distincts from the reference vaccine strains. In both cases, differences were found on locus Bruce07, which had addition of one repeat unit in the case of S19 batches and the deletion of one repeat unit in the case of RB51 batches. In summary, MLVA16 proved to be a molecular tool capable of discriminating small genomic variations and should be included in in vitro official tests. Copyright © 2013. Published by Elsevier Ltd.

  16. Prevention of lethal experimental infection of C57BL/6 mice by vaccination with Brucella abortus strain RB51 expressing Neospora caninum antigens.

    Science.gov (United States)

    Ramamoorthy, Sheela; Sanakkayala, Neelima; Vemulapalli, Ramesh; Duncan, Robert B; Lindsay, David S; Schurig, Gerhart S; Boyle, Stephen M; Kasimanickam, Ramanathan; Sriranganathan, Nammalwar

    2007-11-01

    Bovine abortions caused by the intracellular protozoal parasite Neospora caninum are a major concern to cattle industries worldwide. A strong Th1 immune response is required for protection against N. caninum. Brucella abortus strain RB51 is currently used as a live, attenuated vaccine against bovine brucellosis. Strain RB51 can also be used as an expression vector for heterologous protein expression. In this study, putative protective antigens of N. caninum MIC1, MIC3, GRA2, GRA6 and SRS2, were expressed individually in B. abortus strain RB51. The ability of each of the recombinant RB51 strains to induce N. caninum-specific immunity was assessed in C57BL/6 mice. Mice were immunised by two i.p. inoculations, 4 weeks apart. Five weeks after the second immunisation, spleen cells from the vaccinated mice secreted high levels of IFN-gamma and IL-10 upon in vitro stimulation with N. caninum whole cell lysate antigens. N. caninum-specific antibodies of both IgG1 and IgG2a subtypes were detected in the serum of the vaccinated mice. Mice in the vaccinated and control groups were challenged with 2 x 10(7)N. caninum tachyzoites i.p. and observed for 28 days after vaccination. All unvaccinated control mice died within 7 days. Mice in the MIC1 and GRA6 vaccine groups were completely protected while the mice in the SRS2, GRA2 and MIC3 vaccinated groups were partially protected and experienced 10-50% mortality. The non-recombinant RB51 vector control group experienced an average protection of 69%. These results suggest that expression of protective antigens of N. caninum in B. abortus strain RB51 is a novel approach towards the development of a multivalent vaccine against brucellosis and neosporosis.

  17. Use of the complement fixation and brucellin skin tests to identify cattle vaccinated with Brucella abortus strain RB51.

    Science.gov (United States)

    De Massis, F; Giovannini, A; Di Emidio, B; Ronchi, G F; Tittarelli, M; Di Giannatale, E; Di Ventura, M; Nannini, D; Caporale, V

    2005-01-01

    In the European Union, RB51 vaccine can be used only under strictly controlled conditions for the immunisation of cattle at risk of infection with Brucella abortus. A test is therefore necessary to distinguish vaccinated from unvaccinated animals. The complement fixation test with RB51 antigen (RB51-CFT), dot-blot and gamma-interferon used to identify vaccinated animals have been described, but sensitivity of the tests has been poor and positivity transient after calfhood vaccination. To avail of a rapid and accurate diagnostic tool, the authors produced, controlled and evaluated an experimental brucellin prepared from strain RB51 (RB51 brucellin). The potency of this brucellin was evaluated in guinea-pigs sensitised with RB51 and compared with a commercially available brucellin. Both allergens produced similar biological activity in guinea-pigs. The RB51 brucellin skin test was performed in 10 cattle 414 days after calfhood vaccination with RB51 when they were negative to the RB51-CFT. The skin test revealed 60% sensitivity (with a confidence interval of 95%, CI 30.8%-83.3%) and 100% specificity (CI 60.7%-100%). These findings limit the use of the skin test only for screening to detect RB51 vaccinated herds, not individual animals. Nevertheless, following intradermal inoculation of RB51 brucellin, a transient antibody increase to the RB51-CFT was observed, from day 9 to day 20 post inoculation with RB51 brucellin. This transient antibody increase, when evaluated in parallel with the RB51 brucellin skin test results, enables detection of individual vaccinated animals (sensitivity 100%; CI 76.2%-100%).

  18. An indirect ELISA to detect the serologic response of elk (Cervus elaphus nelsoni) inoculated with Brucella abortus strain RB51.

    Science.gov (United States)

    Colby, Lesley A; Schurig, Gerhardt G; Elzer, Philip H

    2002-10-01

    An indirect enzyme-linked immunosorbent assay (ELISA) was developed to identify elk (Cervus elaphus nelsoni) with Brucella abortus strain RB51 (RB51)-specific antibodies using a mouse monoclonal antibody specific for bovine IgG1. This test was relatively easy to perform, accurate, and easily reproducible; therefore it could be standardized for use between laboratories. In addition, we attempted to compensate for inherent variabilities encountered when comparing ELISA readings from multiple samples taken from many animals over time. Optical density (OD) readings for each sample were converted into a percent positivity value for analysis. A negative cutoff value was determined above which a sample was considered to have a significantly elevated anti-RB51 antibody level. Pre- and postvaccination sera from 64 6-8 mo old elk, divided into four groups (females subcutaneously inoculated with saline (control animals), females ballistically inoculated with RB51, females subcutaneously inoculated with RB51, and males subcutaneously inoculated with RB51) were used. All serum samples were collected between 27 April and 15 November 1995. Values for all saline controls were appropriately below the negative cutoff value. All subcutaneously and ballistically inoculated elk were serologically positive to RB51 for at least two sampling periods during the study. The difference in percent positivity values for the ballistically compared to the subcutaneously inoculated groups was not statistically significant at 8, 10, 14, or 18 wk postvaccination. This suggests that processing RB51 into lactose based pellets and ballistically inoculating elk with these pellets does not alter the detectable elk antibody response. Also, inoculated and control animals can be accurately identified with ELISA at 4-8 weeks postvaccination.

  19. Immune responses of elk to initial and booster vaccinations with Brucella abortus strain RB51 or 19.

    Science.gov (United States)

    Olsen, S C; Fach, S J; Palmer, M V; Sacco, R E; Stoffregen, W C; Waters, W R

    2006-10-01

    Previous studies have suggested that currently available brucellosis vaccines induce poor or no protection in elk (Cervus elaphus nelsoni). In this study, we characterized the immunologic responses of elk after initial or booster vaccination with Brucella abortus strains RB51 (SRB51) and 19 (S19). Elk were vaccinated with saline or 10(10) CFU of SRB51 or S19 (n=seven animals/treatment) and booster vaccinated with a similar dosage of the autologous vaccine at 65 weeks. Compared to nonvaccinates, elk vaccinated with SRB51 or S19 had greater (P0.05) from the responses of nonvaccinated elk. Gamma interferon production in response to autologous or nonautologous Brucella antigens did not differ (P>0.05) between controls and vaccinates after booster vaccination. Flow cytometric techniques suggested that proliferation occurred more frequently in immunoglobulin M-positive cells, with differences between vaccination and control treatments in CD4+ and CD8+ subset proliferation detected only at 22 weeks after initial vaccination. After booster vaccination, one technique ([3H]thymidine incorporation) suggested that proliferative responses to SRB51 antigen, but not S19 antigen, were greater (PBrucella antigens in S19 or SRB51 vaccinates after booster vaccination. Although some cellular immune responses were detected after initial or booster vaccination of elk with SRB51 or S19, our data suggest that responses tend to be transient and much less robust than previously reported in SRB51-vaccinated cattle (Bos taurus) or bison (Bison bison). These data may explain why the vaccination of elk with S19 and SRB51 induces poor protection against brucellosis.

  20. Reduced cerebral infection of Neospora caninum in BALB/c mice vaccinated with recombinant Brucella abortus RB51 strains expressing N. caninum SRS2 and GRA7 proteins.

    Science.gov (United States)

    Vemulapalli, Ramesh; Sanakkayala, Neelima; Gulani, Jatinder; Schurig, Gerhardt G; Boyle, Stephen M; Lindsay, David S; Sriranganathan, Nammalwar

    2007-09-30

    Neospora caninum, an obligate intracellular protozoan parasite, is the causative agent of bovine neosporosis, an important disease affecting the reproductive performance of cattle worldwide. Currently there is no effective vaccine available to prevent N. caninum infection in cattle. In this study, we examined the feasibility of developing a live, recombinant N. caninum vaccine using Brucella abortus vaccine strain RB51 as the expression and delivery vector. We generated two recombinant RB51 strains each expressing SRS2 (RB51/SRS2) or GRA7 (RB51/GRA7) antigens of N. caninum. BALB/c mice immunized by single intraperitoneal inoculation of the recombinant RB51 strains developed IgG antibodies specific to the respective N. caninum antigen. In vitro stimulation of splenocytes from the vaccinated mice with specific antigen resulted in the production of interferon-gamma, but not IL-5 or IL-10, suggesting the development of a Th1 type immune response. Upon challenge with N. caninum tachyzoites, mice vaccinated with strain RB51/SRS2, but not RB51/GRA7, showed significant resistance to cerebral infection when compared to the RB51 vaccinated mice, as determined by the tissue parasite load using a real-time quantitative TaqMan assay. Interestingly, mice vaccinated with either strain RB51 or RB51/GRA7 also contained significantly lower parasite burden in their brains compared to those inoculated with saline. Mice vaccinated with strain RB51/SRS2 or RB51/GRA7 were protected to the same extent as the strain RB51 vaccinated mice against challenge with B. abortus virulent strain 2308. These results suggest that a recombinant RB51 strain expressing an appropriate protective antigen(s), such as SRS2 of N. caninum, can confer protection against both neosporosis and brucellosis.

  1. 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

  2. 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

    Directory of Open Access Journals (Sweden)

    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.

  3. 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.

  4. 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

  5. Immune responses and protection against experimental challenge after vaccination of bison with Brucella abortus strain RB51 or RB51 overexpressing superoxide dismutase and glycosyltransferase genes.

    Science.gov (United States)

    Olsen, S C; Boyle, S M; Schurig, G G; Sriranganathan, N N

    2009-04-01

    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 strain RB51 (RB51) or a recombinant RB51 strain overexpressing superoxide dismutase (sodC) and glycosyltransferase (wboA) genes (RB51+sodC,wboA). Bison were vaccinated with saline only or with 4.6 x 10(10) CFU of RB51 or 7.4 x 10(10) CFU of RB51+sodC,wboA (n = eight animals/treatment). Bison vaccinated with RB51 or RB51+sodC,wboA had greater (P RB51 after vaccination than did nonvaccinates. However, bison vaccinated with RB51+sodC,wboA cleared the vaccine strain from draining lymph nodes faster than bison vaccinated with the parental RB51 strain. Immunologic responses of bison vaccinated with RB51+sodC,wboA were similar to responses of bison vaccinated with RB51. Pregnant bison were intraconjunctivally challenged in midgestation with 10(7) CFU of B. abortus strain 2308. Bison vaccinated with RB51, but not RB51+sodC,wboA vaccinates, had greater protection from abortion, fetal/uterine, mammary, or maternal infection than nonvaccinates. Our data suggest that the RB51+sodC,wboA strain is less efficacious as a calfhood vaccine for bison than the parental RB51 strain. Our data also suggest that the RB51 vaccine is a currently available management tool that could be utilized to help reduce brucellosis in free-ranging bison.

  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. 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

  8. Prevention of vertical transmission of Neospora caninum in C57BL/6 mice vaccinated with Brucella abortus strain RB51 expressing N. caninum protective antigens.

    Science.gov (United States)

    Ramamoorthy, Sheela; Sanakkayala, Neelima; Vemulapalli, Ramesh; Jain, Neeta; Lindsay, David S; Schurig, Gerhardt S; Boyle, Stephen M; Sriranganathan, Nammalwar

    2007-11-01

    Bovine abortions caused by the apicomplexan parasite Neospora caninum have been responsible for severe economic losses to the cattle industry. Infected cows either experience abortion or transmit the parasite transplacentally at a rate of up to 95%. Neospora caninum vaccines that can prevent vertical transmission and ensure disruption in the life cycle of the parasite greatly aid in the management of neosporosis in the cattle industry. Brucella abortus strain RB51, a commercially available vaccine for bovine brucellosis, can also be used as a vector to express plasmid-encoded proteins from other pathogens. Neospora caninum protective antigens MIC1, MIC3, GRA2, GRA6 and SRS2 were expressed in strain RB51. Female C57BL/6 mice were vaccinated with a recombinant strain RB51 expressing N. caninum antigen or irradiated tachyzoites, boosted 4 weeks later and then bred. Antigen-specific IgG, IFN-gamma and IL-10 were detected in vaccinated pregnant mice. Vaccinated mice were challenged with 5 x 10(6)N. caninum tachyzoites between days 11-13 of pregnancy. Brain tissue was collected from pups 3 weeks after birth and examined for the presence of N. caninum by real-time PCR. The RB51-MIC3, RB51-GRA6, irradiated tachyzoite vaccine, pooled strain RB51-Neospora vaccine, RB51-MIC1 and RB51-SRS2 vaccines elicited approximately 6-38% protection against vertical transmission. However, the differences in parasite burden in brain tissue of pups from the control and vaccinated groups were highly significant for all groups. Thus, B. abortus strain RB51 expressing the specific N. caninum antigens induced substantial protection against vertical transmission of N. caninum in mice.

  9. 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

    OpenAIRE

    N.I. Montaña S.; O.E. Rueda L.; C.P. Calderon P.; A. Ortega; A .R. Puentes; M.I. Gallego M.; O.C. Mariño J.

    1998-01-01

    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 contro...

  10. Characterization of large plasmids encoding resistance to toxic heavy metals in Salmonella abortus equi.

    Science.gov (United States)

    Ghosh, A; Singh, A; Ramteke, P W; Singh, V P

    2000-05-27

    Salmonella abortus equi vaccine strains were found to be resistant to high levels of toxic heavy metals--arsenic, chromium, cadmium, and mercury. The two strains 157 and 158 were resistant to ampicillin also. Curing of these strains resulted in loss of one or more resistance marker indicating plasmid borne resistance. Plasmid profile of strain 157 showed presence of three plasmids of 85, 54, and 0.1 Kb, whereas 158 strain showed presence of 85 Kb and 2 Kb plasmids. Plasmids were isolated from strain 157 and introduced into E. coli DH5alpha with a transformation efficiency of 2 x 10(3) transformants/microg DNA. Interestingly the transformants were resistant to antibiotics, heavy metals (As, Cr, Cd, Hg) and was also able to utilize citrate, a trait specific to Salmonella species. We report and establish for the first time the transferable large plasmids encoding resistance to various heavy metals, antibiotics and biochemical nature of S. abortus equi.

  11. Brucella abortus strain RB51 as a vector for heterologous protein expression and induction of specific Th1 type immune responses.

    Science.gov (United States)

    Vemulapalli, R; He, Y; Boyle, S M; Sriranganathan, N; Schurig, G G

    2000-06-01

    Brucella abortus strain RB51 is a stable, rough, attenuated mutant widely used as a live vaccine for bovine brucellosis. Our ultimate goal is to develop strain RB51 as a preferential vector for the delivery of protective antigens of other intracellular pathogens to which the induction of a strong Th1 type of immune response is needed for effective protection. As a first step in that direction, we studied the expression of a foreign reporter protein, beta-galactosidase of Escherichia coli, and the 65-kDa heat shock protein (HSP65) of Mycobacterium bovis in strain RB51. We cloned the promoter sequences of Brucella sodC and groE genes in pBBR1MCS to generate plasmids pBBSODpro and pBBgroE, respectively. The genes for beta-galactosidase (lacZ) and HSP65 were cloned in these plasmids and used to transform strain RB51. An enzyme assay in the recombinant RB51 strains indicated that the level of beta-galactosidase expression is higher under the groE promoter than under the sodC promoter. In strain RB51 containing pBBgroE/lacZ, but not pBBSODpro/lacZ, increased levels of beta-galactosidase expression were observed after subjecting the bacteria to heat shock or following internalization into macrophage-like J774A.1 cells. Mice vaccinated with either of the beta-galactosidase-expressing recombinant RB51 strains developed specific antibodies of predominantly the immunoglobulin G2a (IgG2a) isotype, and in vitro stimulation of their splenocytes with beta-galactosidase induced the secretion of gamma interferon (IFN-gamma), but not interleukin-4 (IL-4). A Th1 type of immune response to HSP65, as indicated by the presence of specific serum IgG2a, but not IgG1, antibodies, and IFN-gamma, but not IL-4, secretion by the specific-antigen-stimulated splenocytes, was also detected in mice vaccinated with strain RB51 containing pBBgroE/hsp65. Studies with mice indicated that expression of beta-galactosidase or HSP65 did not alter either the attenuation characteristics of strain RB51 or

  12. Phenotypic characterization of Lactobacillus strains isolated from ...

    African Journals Online (AJOL)

    Thirty three strains of Lactobacillus were isolated from human milk and infant faeces, animal (cow and goat) milks and from plants (Anagalis arvensis and Bromus mango species). The various strains were identified based on phenotypic tests. Amongst them, 12 strains belonged to group 1, which comprised L. acidophilus, ...

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

    Directory of Open Access Journals (Sweden)

    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.

  14. Brucella abortus detected in cheese from the Amazon region: differentiation of a vaccine strain (B19 from the field strain in the states of Pará, Amapá and Rondônia, Brazil

    Directory of Open Access Journals (Sweden)

    Jacqueline Silva

    Full Text Available Abstract: Brucellosis is an infectious-contagious disease responsible for significant economic losses to the meat and milk supply chain, because it causes reproductive disorders in animals and is a chronic anthropozoonosis. This study was designed to detect the DNA of Brucella spp. in cheese and to differentiate between a vaccine strain (B19 and the field strain. Sixty-six samples of different cheeses which are produced and marketed in three states of the Brazilian Amazon region (Amapá [5 samples], Pará [55 samples] and Rondônia [6 samples] were evaluated. Thirty-nine of these samples were from cheeses made from cow's milk, and 27 were from cheeses made from buffalo milk. Four of the 66 samples were from cheeses produced in milk processing plants regulated by the Federal Inspection Service (Serviço de Inspeção Federal; nine of the samples were from cheeses produced in processing plants regulated by the State Inspection Service (Serviço de Inspeção Estadual; five of the samples were from artisanal cheeses; and the remaining 48 samples were from informally produced cheese. DNA was obtained from the samples following a DNA extraction protocol, and PCR was conducted using primers B4 and B5 to detect Brucella spp. Primers eri1 and eri2 were used to differentiate the field strain from the B19 vaccine strain. The results showed that 21.21% (14/66 of the samples were positive for Brucella spp., of which 21.43% (3/14 were positive for the B. abortus field strain, and 7.14% (1/14 were identified as harboring vaccine strain B19. These results demonstrate that it is possible to identify Brucella spp. in cheese from the Amazon region using the PCR technique and to differentiate the B. abortus field strain from the B19 vaccine strain.

  15. Over-expression of homologous antigens in a leucine auxotroph of Brucella abortus strain RB51 protects mice against a virulent B. suis challenge.

    Science.gov (United States)

    Rajasekaran, Parthiban; Surendran, Naveen; Seleem, Mohamed N; Sriranganathan, Nammalwar; Schurig, Gerhardt G; Boyle, Stephen M

    2011-04-12

    Infection by members of the Gram-negative bacterial genus Brucella causes brucellosis in a variety of mammals. Brucellosis in swine remains a challenge, as there is no vaccine in the USA approved for use in swine against brucellosis. Here, we developed an improved recombinant Brucella abortus vaccine strain RB51 that could afford protection against Brucella suis infection by over-expressing genes encoding homologous proteins: L7/L12 ribosomal protein, Cu/Zn superoxide dismutase [SOD] and glycosyl-transferase [WboA]. Using strain RB51leuB as a platform and an antibiotic-resistance marker free plasmid, strains RB51leuB/SOD, RB51leuB/SOD/L7/L12 and RB51leuB/SOD/WboA were constructed to over-express the antigens: SOD alone, SOD and ribosomal protein L7/L12 or SOD and glycosyl-transferase, respectively. The ability of these vaccine candidates to protect against a virulent B. suis challenge were evaluated in a mouse model. All vaccine groups protected mice significantly (PBrucella antigens can be over-expressed in strain RB51leuB and elicit protective immune responses against brucellosis. Since the plasmid over-expressing homologous antigens does not carry an antibiotic resistance gene, it complies with federal regulations and therefore could be used to develop safer multi-species vaccines for prevention of brucellosis caused by other species of Brucella. Copyright © 2011 Elsevier Ltd. All rights reserved.

  16. Typing of Brucella field strains isolated from livestock populations in Italy between 2001 and 2006

    Directory of Open Access Journals (Sweden)

    Elisabetta Di Giannatale

    2008-06-01

    Full Text Available The identification of species and biovars of Brucella field strains isolated in outbreaks is essential to fully understand the epidemiology of the disease and to trace sources of infection, thereby improving the outcome of brucellosis eradication programmes. It is important to identify the presence of Brucella strains in livestock populations and to determine the presence of new strains that might previously have been considered exotic. In this study, 732 Brucella strains isolated from livestock were submitted for typing by the Italian Istituti Zooprofilattici Sperimentali to the National Reference Laboratory for brucellosis between 2001 and 2006. Animal species affected, biovars typed and spatial distribution of isolates are discussed. Brucella field strains were identified using both conventional bacteriological methods and molecular techniques. Species identification was performed using the AMOS (AbortusMelitensisOvisSuis-polymerase chain reaction. For biovar identification, amplification of omp2a, omp2b and omp31 genes was performed, followed by restriction fragment length polymorphism. Final biovar identification was performed by growth in the presence of basic fuchsin and thionin, using the slide agglutination test with Brucella A- and M- specific antisera.

  17. Comparison of the efficacy of Brucella abortus strain RB51 and Brucella melitensis Rev. 1 live vaccines against experimental infection with Brucella melitensis in pregnant ewes.

    Science.gov (United States)

    el Idrissi, A H; Benkirane, A; el Maadoudi, M; Bouslikhane, M; Berrada, J; Zerouali, A

    2001-12-01

    To test the efficacy of rough Brucella strain vaccines in sheep, a vaccine recently developed in cattle (Brucella abortus strain RB51) was assessed in comparison with the conventional Rev. 1 vaccine. Forty-five ewes from twelve to fourteen months of age, from brucellosis-free flocks, were allotted to three groups of fifteen ewes each. Group one was vaccinated by the conjunctival route with 1.73 x 10(8) colony forming units (CFU) of Rev. 1 vaccine. Group two was vaccinated subcutaneously with 11 x 10(9) CFU of RB51 vaccine and group three was considered as a control. All sheep were challenged at two to three months of gestation with 5 x 10(7) CFU of virulent B. melitensis H38. Vaccination with RB51 vaccine did not result in the production of any antibodies against the O-side chain of lipopolysaccharide, as measured by conventional serological tests (Rose Bengal plate test and complement fixation test). Protection of sheep against abortion and excretion of virulent Brucella strain in vaginal fluid, aborted foetuses and/or non viable lambs at parturition and abortion was significantly lower than that afforded by Rev. 1 vaccine. The difference compared to the control group was not significant. Data from this study suggest that the RB51 vaccine used for cattle vaccination does not provide effective protection of sheep against abortion induced by B. melitensis.

  18. Serologic responses, biosafety and clearance of four dosages of Brucella abortus strain RB51 in 6-10 months old water buffalo (Bubalus bubalis).

    Science.gov (United States)

    Diptee, M D; Adesiyun, A A; Asgarali, Z; Campbell, M; Adone, R

    2006-01-15

    Thirty water buffalo were obtained from a brucellosis-free farm in order to evaluate antibody responses, bacterial clearance and safety to Brucella abortus strain RB51 vaccine in a dose response study. The animals were randomly divided into five treatment groups. Groups I-V received the recommended dose of RB51 vaccine (RD) once, RD twice 4 weeks apart, double RD once, double RD twice 4 weeks apart and saline once, respectively. Antibody responses to RB51 were monitored at 2, 4, 6, 8, 10, 12, 16 18, 22, 24 and 27 post-initial-inoculation weeks (PIW). Clearance of RB51 from the prescapular lymph node was evaluated at 2, 4, 6, 12, 18 and 24 PIW for groups 1, III and V and at 6, 8, 10, 16, 22 and 27 PIW for groups II and IV. To evaluate shedding of the RB51 strain, nasal, conjunctival, vaginal or preputial swabs were taken from all experimental animals at 1, 2, 3, 4, 6, 8 and 12 PIW. Sera taken at all PIW were negative for field strain B. abortus by both the buffered plate agglutination test (BPAT) and competitive enzyme-linked immunosorbent assay (c-ELISA). Antibody responses to RB51 were demonstrated in all vaccinates but not in the controls, up to 12 PIW, by complement fixation test (CFT) and the dot-blot assay with an 83.7% agreement for both tests. Clearance of RB51 occurred between 6 and 12 PIW in group I but less than 2 weeks after booster vaccinations in groups II and IV and between 4 and 6 PIW in group III. RB51 was not recovered at any time from swabs obtained from either RB51-vaccinates or non-vaccinates. The results of this study indicate that serologic responses to RB51 vaccination can be monitored by both CFT and dot-blot assay in water buffalo. Our data also indicates that RB51 vaccination does not interfere with brucellosis sero-surveillance and is safe (no serological and bacteriological evidence of spread to non-vaccinates, no adverse clinical signs or detectable abnormalities on haematology and serum biochemistry) for use in water buffalo.

  19. Update: potential exposures to attenuated vaccine strain Brucella abortus RB51 during a laboratory proficiency test--United States and Canada, 2007.

    Science.gov (United States)

    2008-01-18

    In November 2007, New York State Department of Health (NYSDOH) officials notified CDC of potential exposures to attenuated vaccine strain Brucella abortus RB51 (RB51) in multiple clinical laboratories that participated in a Laboratory Preparedness Survey (LPS) proficiency test. NYSDOH conducted a survey of participating laboratories and identified 17 laboratories that reported handling the RB51 sample in a manner placing lab workers at potential risk for exposure. Subsequently, CDC recommended that public health officials conduct a review of biosafety practices at all LPS-participating laboratories to identify any additional RB51 exposures. This report summarizes the results of investigations in 36 states, two cities, one county, and the District of Columbia. As of January 14, 2008, follow-up by public health officials with LPS-participating laboratories throughout the United States identified a total of 916 laboratory workers in 254 laboratories with potential RB51 exposure. The results highlight the need for routine adherence to recommended biosafety practices when working with infectious organisms, particularly during widespread infectious-disease events, including bioterrorism attacks.

  20. Immunologic responses of bison to vaccination with Brucella abortus strain RB51: comparison of parenteral to ballistic delivery via compressed pellets or photopolymerized hydrogels.

    Science.gov (United States)

    Olsen, Steven C; Christie, R J; Grainger, D W; Stoffregen, W S

    2006-02-27

    This study compared responses of bison calves to 10(10)CFU of Brucella abortus strain RB51 (SRB51) delivered by parenteral or ballistic methods. Two types of biobullet payloads were evaluated; compacted SRB51 pellets or SRB51 encapsulated in photopolymerized poly(ethylene glycol) hydrogels. Bison were vaccinated with saline, parenteral SRB51 alone, or in combination with Spirovac, or ballistically with compressed SRB51 or hydrogel biobullets. Bison parenterally vaccinated with SRB51 had greater (P<0.05) immunologic responses when compared to control bison. Co-administration of Spirovac as an adjuvant did not influence immunologic responses. As compared to compressed SRB51 biobullets, ballistic vaccination with hydrogel biobullets increased cellular immune responses at some sampling times. Our data suggest that hydrogel formulations of SRB51 may be a superior alternative to compressed SRB51 tablets for ballistic vaccination of bison. Although preliminary, data suggests that immunologic responses of bison to SRB51 hydrogel bullets are similar to responses after parenteral vaccination with SRB51.

  1. Defensin susceptibility and colonization in the mouse model of AJ100, a polymyxin B-resistant, Brucella abortus RB51 isolate.

    Science.gov (United States)

    Halling, Shirley M; Jensen, Allen E; Olsen, Steven C

    2008-03-01

    Intracellular pathogens selected for increased susceptibility to polycations are commonly attenuated, yet the effect of decreased susceptibility to polycations on pathogenicity has not been researched. The polymyxin-resistant mutant Brucella abortus AJ100 was characterized by comparing its susceptibility to the polycationic antibiotic polymyxin B, defensins, and lactoferricin, and its colonization and clearance in the mouse model to the parent strain RB51. MIC (minimum inhibitory concentration) values determined by Etest for AJ100 and RB51 were 1.5 and 0.25 mug/ml, respectively. Though AJ100 is less susceptible to polymyxin B than RB51, it was more susceptible than its parent strain to the cationic defensins melittin, magainin 2, and cecropin P1. In the mouse model, initial colonization of the spleen was lower for AJ100 than RB51, and the rate of clearance from the spleen was faster for AJ100 than RB51. However, initial colonization and clearance rates of AJ100 from the liver were indistinguishable from those of RB51. This study suggests that the susceptibility profile of Brucella to polycationic defensins rather than polymyxin B may be indicative of differential survival in the spleen and liver in the mouse and is indicative of spleen and liver residential macrophages' differing ability to inactivate Brucella.

  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

    Directory of Open Access Journals (Sweden)

    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. Shewanella strain isolated from black powder

    Energy Technology Data Exchange (ETDEWEB)

    Lutterbach, Marcia T.S.; Contador, Luciana S.; Oliveira, Ana Lucia C.; Galvao, Mariana M. [National Institute of Technology (INT), Rio de Janeiro, RJ (Brazil); Pimenta, Gutemberg S. [PETROBRAS, Rio de Janeiro, RJ (Brazil)

    2009-07-01

    Black powder is a term frequently used to refer to residues formed by various types of iron sulfides mixed with contaminants eventually present in the natural gas flow. According to some researchers, the occurrence of black powder in gas pipelines, besides its chemical corrosion origin, can be directly related to the sulfate-reducing bacteria (SRB) metabolism in this environment. A black powder sample was inoculated in a Post gate E medium modified with the addition of thioglycolate. The resulting positive culture was kept in the laboratory for four years until its use. A dilution technique was then performed aiming to isolate an SRB strain. The bacterial strain isolated and identified through DNA sequencing was not an SRB but rather a Shewanella sp. Compared to the sulfate-reducing bacteria group-traditionally considered the foremost responsible for microbially-influenced corrosion (MIC) - Shewanella is a facultative anaerobe and has a versatile metabolism. Shewanella is able to reduce ferric iron and sulfite, oxidize hydrogen gas, and produce hydrogen sulfide; therefore, these bacteria can be responsible for MIC and pit formation. The isolated Shewanella was used in a corrosion experiment, and the corrosion products were characterized by X-ray diffraction, identifying iron sulfides, iron oxides, and sulfur. Our results indicate that the strain isolated, S. putrefaciens, plays a key role in corrosion problems in gas pipelines. (author)

  4. 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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. 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...

  6. 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

    OpenAIRE

    M. Villarroel; M. Grell; R. Saenz

    2000-01-01

    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 ...

  7. 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

    OpenAIRE

    M Mancilla; M Villarroel; ME Saldías; J Soto; AM Zárraga

    2008-01-01

    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....

  8. Multi-locus variable-number tandem repeat analysis of Chinese Brucella strains isolated from 1953 to 2013.

    Science.gov (United States)

    Tian, Guo-Zhong; Cui, Bu-Yun; Piao, Dong-Ri; Zhao, Hong-Yan; Li, Lan-Yu; Liu, Xi; Xiao, Pei; Zhao, Zhong-Zhi; Xu, Li-Qing; Jiang, Hai; Li, Zhen-Jun

    2017-05-02

    Brucellosis was a common human and livestock disease caused by Brucella strains, the category B priority pathogens by the US Center for Disease Control (CDC). Identified as a priority disease in human and livestock populations, the increasing incidence in recent years in China needs urgent control measures for this disease but the molecular background important for monitoring the epidemiology of Brucella strains at the national level is still lacking. A total of 600 Brucella isolates collected during 60 years (from 1953 to 2013) in China were genotyped by multiple locus variable-number tandem repeat analysis (MLVA) and the variation degree of MLVA11 loci was calculated by the Hunter Gaston Diversity Index (HGDI) values. The charts and map were processed by Excel 2013, and cluster analysis and epidemiological distribution was performed using BioNumerics (version 5.1). The 600 representative Brucella isolates fell into 104 genotypes with 58 singleton genotypes by the MLVA11 assay, including B. melitensis biovars 2 and 3 (five main genotypes), B. abortus biovars 1 and 3 (two main genotypes), B. suis biovars 1 and 3 (three main genotypes), and B. canis (two main genotypes) respectively. While most B. suis biovar 1 and biovar 3 were respectively found in northern provinces and southern provinces, B. melitensis and B. abortus strains were dominant in China. Canine Brucellosis was only found in animals without any human cases reported. Eight Brucellosis epidemic peaks emerged during the 60 years between 1953 and 2013: 1955 - 1959, 1962 - 1969, 1971 - 1975, 1977 - 1983, 1985 - 1989, 1992 - 1997, 2000 - 2008 and 2010 - 2013 in China. Brucellosis has its unique molecular epidemiological patterns with specific spatial and temporal distribution according to MLVA. IDOP-D-16-00101.

  9. Enrichment and isolation of microbial strains degrading bioplastic ...

    African Journals Online (AJOL)

    Polyvinyl alcohol (PVA) degrading bacterial strains were isolated from various environmental sites rich in plastic wastes by using the enrichment culture technique. Among the various isolated strains, the selected potent PVA degrading bacterial strains were tentatively characterized as Bacillus and Pseudomonas sp.

  10. Enrichment and isolation of microbial strains degrading bioplastic ...

    African Journals Online (AJOL)

    acer

    2015-07-08

    Jul 8, 2015 ... Polyvinyl alcohol (PVA) degrading bacterial strains were isolated from various environmental sites rich in plastic wastes by using the enrichment culture technique. Among the various isolated strains, the selected potent PVA degrading bacterial strains were tentatively characterized as Bacillus and.

  11. 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. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  12. Genetic variations among Mycoplasma bovis strains isolated from Danish cattle

    DEFF Research Database (Denmark)

    Kusiluka, L.J.M.; Kokotovic, Branko; Ojeniyi, B.

    2000-01-01

    The genetic heterogeneity of Mycoplasma bovis strains isolated in Denmark over a 17-year period was investigated. Forty-two field strains isolated from different geographic locations and specimens, including strains from 21 herds involved in two outbreaks of M. bovis-induced mastitis, and the type....... Among the analyzed strains, 18 different AFLP profiles were detected. The similarity between individual fingerprints, calculated by Dice similarity coefficient, ranged from 0.9 to 1.0. Twenty-five strains, including 23 which were isolated during two outbreaks of M. bovis-induced mastitis which occurred...... 2 years apart; showed indistinguishable AFLP patterns. More genetic diversify was observed among the recent strains. The similarity of the genotypes of the field strains to that of the M. bovis type strain (PG45(T)) was 97.7%. The results of this study have demonstrated a remarkable genomic...

  13. Fluorene biodegradation potentials of Bacillus strains isolated from ...

    African Journals Online (AJOL)

    Fluorene biodegradation potentials of Bacillus strains isolated from tropical hydrocarbon-contaminated soils. ... The rate of degradation of fluorene (50 mg/L) by the two isolates, after 30 days of incubation were 0.09 and 0.08 mg/L/h for strains BM1 and BR1, respectively. Gas chromatographic analyses of residual fluorene, ...

  14. Isolation of polyvinyl chloride degrading bacterial strains from ...

    African Journals Online (AJOL)

    In the present investigation, potent microbial strains degrading plastic constituting polymer polyvinyl chloride (PVC) were isolated using enrichment culture technique. To increase the chances of isolating such strain which could have adapted to metabolize plastic constituting polymers, samples were collected from different ...

  15. Aflatoxin B1 producing potential of Aspergillus flavus strains isolated ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-20

    Jul 20, 2009 ... Extraction of AFB1 from A. flavus strains grown on agar media. Eighty five strains of A. flavus isolated from rice grain samples were grown on sterilized different agar media (AFPA, Czapeks agar, PDA and YES agar) for 5 days at 25 ± 2°C. Three replications were maintained for each isolate for each media.

  16. Antibiotics susceptibility pattern of Escherichia coli strains isolated ...

    African Journals Online (AJOL)

    This study was carried out to determine the susceptibility of E coli strains isolated from broiler and layer chicken with colisepticemia to antibiotics used in poultry industry in the country. A total of fifty seven E. coli strains isolated from 43 broiler and14 layer farms with colisepticemia in Khartoum and Gezera state were ...

  17. Identification and determination of antibiotic susceptibilities of Brucella strains isolated from patients in van, Turkey by conventional and molecular methods.

    Science.gov (United States)

    Parlak, Mehmet; Güdücüoğlu, Hüseyin; Bayram, Yasemin; Çıkman, Aytekin; Aypak, Cenk; Kılıç, Selçuk; Berktaş, Mustafa

    2013-01-01

    Brucellosis is a worldwide zoonotic disease and still constitutes a major public health problem. In this study, we aimed to identify biovars of Brucella strains isolated from clinical specimens taken from brucellosis patients from the Eastern Anatolia region as well determine the susceptibility of these isolates to tigecycline and azithromycin, drugs that may serve as alternatives to the conventional drugs used in the therapy. Seventy-five Brucella spp. isolates were included in the study. All strains were identified by both conventional and molecular methods. Brucella Multiplex PCR kit (FC-Biotech, Code: 0301, Turkey) and B. melitensis biovar typing PCR kit (FC-Biotech, Code: 0302, Turkey) were used for molecular typing. Antimicrobial susceptibilities of all strains were determined by E-tests. By conventional biotyping, 73 strains were identified as B. melitensis biovar 3 and two strains as B. abortus biovar 3. Molecular typing results were compatible with conventional methods. The MIC50 and MIC90 values of doxycycline were 0.047 and 0.094; tigecycline 0.094 and 0.125; trimethoprim/sulfamethoxazole 0.064 and 0.19; ciprofloxacin 0.19 for both; streptomycin 0.75 and 1; rifampin 1 and 2 and azithromycin 4 and 8. According to the MIC values, doxycycline was found to be the most effective antibiotic, followed by tigecycline, trimethoprim-sulfamethoxazole and ciprofloxacin. Currently recommended antibiotics for the treatment of brucellosis such as doxycycline, rifampin, streptomycin, trimethoprim-sulfamethoxazole and ciprofloxacin were found to be still effective. While our results showed that tigecycline can be used an alternative agent in the treatment of brucellosis, azithromycin has not been confirmed as an appropriate agent for the treatment.

  18. Antimicrobial resistance of bacterial strains isolated from orange ...

    African Journals Online (AJOL)

    ... obtained for the strains of S. aureus. The MIC for cloxacillin was 0.1-1.0mg/ml for E. coli strains, and 0.01-1.0mg/ml for S. aureus strains. In all, ten strains of the bacterial isolates had evidence for the production of β-lactamases. Key words: Orange juice, antibiotics, resistance pattern, β-lactamase, microbiological standard.

  19. 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. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  20. [Isolation and identification of Cronobacter (Enterobacter sakazakii) strains from food].

    Science.gov (United States)

    Dong, Xiaohui; Li, Chengsi; Wu, Qingping; Zhang, Jumei; Mo, Shuping; Guo, Weipeng; Yang, Xiaojuan; Xu, Xiaoke

    2013-05-04

    This study aimed to detect and quantify Cronobacter in 300 powdered milk samples and 50 non-powdered milk samples. Totally, 24 Cronobacter (formerly Enterobacter sakazakii) strains isolated from powdered milk and other foods were identified and confirmed. Cronobacter strains were detected quantitatively using most probable number (MPN) method and molecular detection method. We identified 24 Cronobacter strains using biochemical patterns, including indole production and dulcitol, malonate, melezitose, turanose, and myo-Inositol utilization. Of the 24 strains, their 16S rRNA genes were sequenced, and constructed phylogenetic tree by N-J (Neighbour-Joining) with the 16S rRNA gene sequences of 17 identified Cronobacter strains and 10 non-Cronobacter strains. Quantitative detection showed that Cronobacter strains were detected in 23 out of 350 samples yielding 6.6% detection rate. Twenty-four Cronobacter strains were isolated from 23 samples and the Cronobacter was more than 100 MPN/100g in 4 samples out of 23 samples. The 24 Cronobacter spp. isolates strains were identified and confirmed, including 19 Cronobacter sakazakii strains, 2 C. malonaticus strains, 2 C. dubliensis subsp. lactaridi strains, and 1 C. muytjensii strain. The combination of molecular detection method and most probable number (MPN) method could be suitable for the detection of Cronobacter in powdered milk, with low rate of contamination and high demand of quantitative detection. 24 isolated strains were confirmed and identified by biochemical patterns and molecular technology, and C. sakazakii could be the dominant species. The problem of Cronobacter in powdered milk should be a hidden danger to nurseling, and should catch the government and consumer's attention.

  1. Male rats transmit Brucella abortus biotype 1 through sexual intercourse.

    Science.gov (United States)

    Islam, Md Ariful; Khatun, Mst Minara; Baek, Byeong-Kirl

    2013-08-30

    The aim of this study was to evaluate transmission of Brucella abortus biotype 1 via sexual intercourse in rats. Male and female virgin Sprague-Dawley (SD) rats were experimentally infected intraperitoneally with 1×10(9)colony forming units (CFU) of B. abortus biotype 1, a Korean bovine isolate. At 14 days after infection, infected male rats (n=10) were housed with uninfected female rats (n=10) and infected female rats (n=10) were housed with uninfected male rats (n=10) for a period of one month. During this period all uninfected female rats became pregnant and 6 of 10 infected female rats became pregnant. Serum from two out of 10 female uninfected rats had positive reactions in the Rose Bengal Plate Agglutination Test (RBPAT), Tube Agglutination Test (TAT) or the Enzyme-Linked Immunosorbent Assay (ELISA); whereas none of the uninfected male rat had positive reactions in these tests. Using bacteriological culture and AMOS-PCR assay, B. abortus biotype 1 was isolated and identified from two uninfected female rats and all of the uninfected male rats were found negative for B. abortus biotype 1. It was concluded that transmission of B. abortus biotype 1 from infected male to uninfected female rats resulted from sexual intercourse. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Isolation, identification and characterization of regional indigenous Saccharomyces cerevisiae strains.

    Science.gov (United States)

    Šuranská, Hana; Vránová, Dana; Omelková, Jiřina

    2016-01-01

    In the present work we isolated and identified various indigenous Saccharomyces cerevisiae strains and screened them for the selected oenological properties. These S. cerevisiae strains were isolated from berries and spontaneously fermented musts. The grape berries (Sauvignon blanc and Pinot noir) were grown under the integrated and organic mode of farming in the South Moravia (Czech Republic) wine region. Modern genotyping techniques such as PCR-fingerprinting and interdelta PCR typing were employed to differentiate among indigenous S. cerevisiae strains. This combination of the methods provides a rapid and relatively simple approach for identification of yeast of S. cerevisiae at strain level. In total, 120 isolates were identified and grouped by molecular approaches and 45 of the representative strains were tested for selected important oenological properties including ethanol, sulfur dioxide and osmotic stress tolerance, intensity of flocculation and desirable enzymatic activities. Their ability to produce and utilize acetic/malic acid was examined as well; in addition, H2S production as an undesirable property was screened. The oenological characteristics of indigenous isolates were compared to a commercially available S. cerevisiae BS6 strain, which is commonly used as the starter culture. Finally, some indigenous strains coming from organically treated grape berries were chosen for their promising oenological properties and these strains will be used as the starter culture, because application of a selected indigenous S. cerevisiae strain can enhance the regional character of the wines. Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  3. Isolation, identification and characterization of regional indigenous Saccharomyces cerevisiae strains

    Directory of Open Access Journals (Sweden)

    Hana Šuranská

    2016-03-01

    Full Text Available Abstract In the present work we isolated and identified various indigenous Saccharomyces cerevisiae strains and screened them for the selected oenological properties. These S. cerevisiae strains were isolated from berries and spontaneously fermented musts. The grape berries (Sauvignon blanc and Pinot noir were grown under the integrated and organic mode of farming in the South Moravia (Czech Republic wine region. Modern genotyping techniques such as PCR-fingerprinting and interdelta PCR typing were employed to differentiate among indigenous S. cerevisiae strains. This combination of the methods provides a rapid and relatively simple approach for identification of yeast of S. cerevisiae at strain level. In total, 120 isolates were identified and grouped by molecular approaches and 45 of the representative strains were tested for selected important oenological properties including ethanol, sulfur dioxide and osmotic stress tolerance, intensity of flocculation and desirable enzymatic activities. Their ability to produce and utilize acetic/malic acid was examined as well; in addition, H2S production as an undesirable property was screened. The oenological characteristics of indigenous isolates were compared to a commercially available S. cerevisiae BS6 strain, which is commonly used as the starter culture. Finally, some indigenous strains coming from organically treated grape berries were chosen for their promising oenological properties and these strains will be used as the starter culture, because application of a selected indigenous S. cerevisiae strain can enhance the regional character of the wines.

  4. Method for bacteriophage isolation against target Campylobacter strains

    National Research Council Canada - National Science Library

    Carvalho, C; Susano, M; Fernandes, E; Santos, S; Gannon, B; Nicolau, A; Gibbs, P; Teixeira, P; Azeredo, J

    2010-01-01

    .... This work focuses on the isolation of Campylobacter coli lytic bacteriophages (phages) against target C. coli strains. A method involving the enrichment of free-range chicken samples in a broth containing the target C...

  5. Identification of a new strain of Actinomadura isolated from Saharan ...

    African Journals Online (AJOL)

    Identification of a new strain of Actinomadura isolated from Saharan soil and ... soil, with strong antifungal activity against pathogenic and toxinogenic fungi, was ... The butanolic extract contained four bioactive spots detected on thin layer ...

  6. Reclassification of 30 Pasteurellaceae strains isolated from rodents.

    Science.gov (United States)

    Boot, R; Bisgaard, M

    1995-07-01

    Thirty Pasteurellaceae strains isolated from gerbil, guineapig, hamster, mouse, muskrat and rat were reinvestigated and reclassified after comparison with reference strains. Strains originally described as Pasteurella pneumotropica were reclassified as [Pasteurella] pneumotropica Heyl biotype (7), [P.] pneumotropica Jawetz biotype (1), Pasteurella dagmatis (1) or Taxon 22 (2). Strains previously reported as Actinobacillus sp. were reclassified as [P.] pneumotropica biotype Jawetz (3), P. dagmatis (3) or Taxon 6 (7). Strains earlier described as Pasteurella gallinarum were renamed as SP group pasteurella (4) or Taxon 25 (2). Some of these reclassified Pasteurellaceae have not been reported previously in rodents. The present findings underline the importance of extended characterization of isolates and comparison with references strains to avoid misclassification within the family Pasteurellaceae Pohl 1981.

  7. Taxonomy of Streptomyces strains isolated from rhizospheres of ...

    African Journals Online (AJOL)

    This work was designed to identify and taxonomically classify Streptomyces strains isolated from the rhizospheres of various plant species; banana, rose, pomegranate and grape plants, having antagonistic activity against some microbial (bacteria and fungi) tissue culture contaminants. Streptomyces strains with the most ...

  8. Antimicrobial Resistance of Staphylococcal Strains Isolated from Various Pathological Products

    Directory of Open Access Journals (Sweden)

    Laura-Mihaela SIMON

    2010-12-01

    Full Text Available Background: The optimal choice of antimicrobial therapy is an important problem in hospital environment in which the selection of resistant and virulent strains easy occurs. S. aureus and especially MRSA(methicillin-resistant S. aureus creates difficulties in both treatment and prevention of nosocomial infections. Aim: The purpose of this study is to determine the sensitivity and the resistance to chemotherapy of staphylococci strains isolated from various pathological products. Material and Method: We identified Staphylococccus species after morphological appearance, culture properties, the production of coagulase, hemolisines and the enzyme activity. The susceptibility tests were performed on Mueller-Hinton medium according to CLSI (Clinical and Laboratory Standards Institute. Results: The strains were: MSSA (methicillin-susceptible S. aureus (74%, MRSA (8%, MLS B (macrolides, lincosamides and type B streptogramines resistance (12% and MRSA and MLS B (6%. MRSA strains were more frequently isolated from sputum. MRSA associated with the MLS B strains were more frequently isolated from pus. MLS B strains were more frequently isolated from sputum and throat secretions. All S. aureus strains were susceptible to vancomycin and teicoplanin. Conclusions: All staphylococcal infections require resistance testing before treatment. MLS B shows a high prevalence among strains of S. aureus. The association between MLS B and MRSA remains a major problem in Romania.

  9. Rat islet isolation yield and function are donor strain dependent

    NARCIS (Netherlands)

    de Groot, M; de Haan, BJ; Schuurs, TA; van Schilfgaarde, R; Leuvenink, HGD; KEIZER, J

    Effective rat islet isolation is pertinent for successful islet transplantation and islet studies in vitro. To determine which rat strain yields the highest number of pure and functional islets, four commonly used rat strains were compared with regard to islet yield, islet purity and islet function.

  10. Isolation and characterization of lipase-producing Bacillus strains ...

    African Journals Online (AJOL)

    Bacillus strains (B1 - B5) producing extra cellular lipase were isolated from the soil sample of coconut oil industry. The strains were identified by morphological and biochemical characters. Growth of the organisms and lipase production were measured with varying pH (4 - 9) temperature (27, 37 and 47ºC) and various ...

  11. Preliminary characterization of Rhizobium strains isolated from ...

    African Journals Online (AJOL)

    SERVER

    2008-03-18

    Mar 18, 2008 ... range of carbohydrates and salts of organic acids as carbon sources which are also diagnostic for root nodule bacteria (Sa et al., 1993; Rodriguez-Navarro et al., 2000;. Zerhari et al., 2000). Utilizasation of different compounds by strains, as sole carbon and nitrogen sources is one of the most useful traits for ...

  12. Ethanol production potential of local yeast strains isolated from ripe ...

    African Journals Online (AJOL)

    The ability of different yeast strains isolated from ripe banana peels to produce ethanol was investigated. Of the 8 isolates screened for their fermentation ability, 5 showed enhanced performance and were subsequently identified and assessed for important ethanol fermentation attributes such as ethanol producing ability, ...

  13. Identification of Candida strains isolated from Tanzanian pregnant ...

    African Journals Online (AJOL)

    Objective: To identify Candida strains isolated from Tanzanian women (13 to 45 years) with vaginal candidiasis. Design: A cross-sectional study. Setting: Antenatal clinic in llala district hospital in Dar es Salaam, Tanzania from March 1998 to December 2000. Results: The identities of the 272 isolates tested with API Candida ...

  14. Characterization of Rhizobium strain isolated from the roots of ...

    African Journals Online (AJOL)

    Trigonella foenumgraecum (fenugreek) is known for its dietary protein source, medicinal properties and symbiotic nitrogen fixation by Rhizobium present in its root nodules. The present study describes the characterization of a Rhizobium strain isolated from root nodules of fenugreek. The Rhizobium isolates were rod ...

  15. Heavy metal resistance by two bacteria strains isolated from a ...

    African Journals Online (AJOL)

    USER

    2010-06-28

    Jun 28, 2010 ... essential but poisonous for plants, animals and humans. It has high toxicity ... were used to isolate cadmium and zinc tolerant bacteria strains, respectively. 0.1 ml of the liquid soil bacterial supernatant was plated on each disk. ..... isolated from root nodule of Lespedeza cuneata in gold mine tailings in China ...

  16. Antimicrobial drug susceptibility of Neisseria meningitidis strains isolated from carriers

    Directory of Open Access Journals (Sweden)

    Dayamí García

    2000-06-01

    Full Text Available When it is necessary to determine the susceptibility of Neisseria meningitidis (Nm strains to antimicrobial drugs, it is important to consider that it should be analyzed in a double context. One of them related to the use of drugs in a specific medical treatment; and the other; to chemoprophylatic drugs, both with the same purpose: the accurate selection of the “in vivo” antimicrobial agent. This requires the study of the sensitivity and resistance of strains isolated in both carriers and patients. With the aim of further studying the behavior of the strains that currently circulate in Cuba, an antimicrobial drug susceptibility study was conducted in 90 strains isolated from carriers during the first half of 1998. The agar dilution method was used to determine the minimum inhibitory concentrations (MICs to: penicillin, ampicillin, rifampin, sulfadiazine, chloramphenicol, ciprofloxacin, ceftriaxone, cefotaxime. The study of the three latter drugs was done for the first time in our country. The search for β- lactamase-producer strains was also performed. There was a predominance of penicillin sensitive strains (82,2% with an intermediate sensitivity to ampicillin (57,8%, while 70% of the strains were sensitive to sulfadiazine. Regarding the rest of the antimicrobial drugs, 100% of the strains were sensitive. The paper shows the MICs for each drug as well as the phenotypic characteristics of the strains with the penicillin and sulfadiazine sensitivity and resistance patterns. No β-lactamase-producer strains were found.

  17. Biofilm production among Staphylococcus epidermidis strains isolated from healthy people

    Directory of Open Access Journals (Sweden)

    Fateh Rahimi

    2016-09-01

    Full Text Available Introduction: Staphylococcus epidermidis is well documented as a nosocomial pathogen causing biofilm in patients and healthy people. The aim of this study was to analyze the biofilm formation of S. epidermidis strains isolated from healthy people during 2013-2014. Materials and methods: Totally 200 healthy people were selected and the sampling was carried out from arm, armpit and axillary area using sterile swaps. Swaps were transferred to thiogylcollate broth and then cultured on mannitol salt agar plates. Isolates were identified at the species level using biochemical tests. Potential of biofilm formation of strains was measured using congo red agar plate and microtiter plate tests. Genes involved in biofilm formation, icaA and icaD, were detected using PCR. Results: Totally 104 S. epidermidis strains were isolated from healthy people. Amongst these, 66 (63% and 38 (37% strains were positive and negative for biofilm formation, respectively. icaA and icaD genes were detected in 100% of strains. Discussion and conclusion: Prevalence of biofilm producing S. epidermidis isolates among healthy people indicating their colonization with hospital strains. Prevalence of such strains is  urgent for public health.

  18. Hexadecane degradation by bacterial strains isolated from ...

    African Journals Online (AJOL)

    A study was undertaken to detect and monitor the degradation of hexadecane by three potential degrading bacteria (Pseudomonas putida, Rhodococcus erythroplolis and Bacillus thermoleovorans) isolated from contaminated soils in Riyadh, Saudi Arabia. The extraction of the bacterial populations from these polluted soils ...

  19. Antimicrobial susceptibility of Salmonella enterica strains isolated ...

    African Journals Online (AJOL)

    SARAH

    2015-11-30

    Nov 30, 2015 ... ABSTRACT. Objectives: The aim of this study was to determine the prevalence and antibiotic resistance profile of. Salmonella enterica isolated from raw beef, mutton and intestines sold in Ouagadougou; Burkina Faso. Methodology and Results: A total of 450 samples from raw meat of beef (n=175), mutton ...

  20. A study of V. cholerae strains isolated in Bombay.

    Directory of Open Access Journals (Sweden)

    Saraswathi K

    1990-07-01

    Full Text Available Of 935 faecal samples studied over a period of one year, V. cholerae 01 was isolated from 102 samples (10.9%. All the strains were found to be E1 Tor Ogawa. The strains belonging to the phage types 2 and 4 were encountered in our study, type 2 being the highest (76.5%. The sensitivity pattern of all strains to the commonly used antibiotics was determined. Strains sensitive to gentamicin (92.2%, nalidixic acid (85.3%, kanamycin (83.3%, cotrimoxazole (80.4% and chloramphenicol (75.5% were observed. Out of the total, 36.3%, 29.4% and 28.4% of V. cholerae strains were found to be resistant to ampicillin, streptomycin and tetracycline respectively. V. cholerae was isolated throughout the year indicating the endemicity of cholera in Bombay.

  1. Neutralization epitope patterns of poliovirus strains isolated from paralytic cases.

    Science.gov (United States)

    Crainic, R; Blondel, B; Aubert-Combiescu, A; Beytout, D; Couillin, P; Candrea, A; Boué, A; Horaud, F

    1984-01-01

    Monoclonal antibodies were used in previous work to define neutralization epitopes (NE) of poliovirus during its natural variation upon interhuman passages. Two kind of NE were found: constant (K) epitopes, present on practically all poliovirus strains of the same serotype, and variable (V) epitopes, which were strain-specific either for wild-laboratory strains (Mahoney, MEF1, and Saukett) (VW epitopes), or for attenuated Sabin strains (VS epitopes), or for attenuated Sabin strains (VS epitopes). In the present paper we analyzed the NE formulae of poliovirus strains of each of the three serotypes isolated from paralytic cases that have occurred after mass vaccination with oral poliovaccine (Sabin). Type 1 isolates from both the central nervous system and stools showed the existence of the four VW epitopes characteristic for Mahoney virus. However, in some cases, variation of the K epitope formula was noticed. As concerns the paralytic cases where type 2 poliovirus was isolated, the situation was more complex due to the existence of three different VS epitopes. Various patterns of VS epitopes were encountered, making difficult the definition of the strains origin. Most of the type 3 strains isolated from paralytic cases contained one or both VS epitopes, some of them having an identical NE formula to the Sabin type 3 vaccine virus. These data showed that the use of strain-specific neutralizing monoclonal antibodies for the characterization of strain origin is more complex than generally agreed. The multitude of NE coexisting on the same virus particle and their variation during viral replication in the human intestine call for cautious interpretation of the results obtained in poliovirus intratypic differentiation by neutralizing monoclonal antibodies.

  2. Isolation of Mycoplasma capricolum-like strains from chickens.

    Science.gov (United States)

    Bencina, Dusan; Bradbury, Janet M; Stipkovits, Laszlo; Varga, Zsuzsana; Razpet, Andrej; Bidovec, Andrej; Dovc, Peter

    2006-01-10

    Members of the genus Mycoplasma infect a wide range of hosts, but individual Mycoplasma species tend to exhibit a considerable degree of host specificity. We characterized Mycoplasma strain 700, isolated from a kidney of a layer hen in Spain and Mycoplasma strains ULB-A and ULB-B, isolated from the air sac and from the bile of stunted broiler chickens in Slovenia. The serologic examination showed that these three strains are antigenically unrelated to all of the recognized Mycoplasma species of avian origin, but closely related to the ruminant mycoplasma Mycoplasma capricolum subspecies capricolum (M. capricolum). The comparison of their 16S rRNA gene sequences with the sequence of M. capricolum (California kid) revealed 99.66% sequence identity for the strain 700 and 99.59% identity for strains ULB-A and ULB-B. Moreover, the predicted DnaK sequences of the M. capricolum-like strains, isolated from chickens, were identical to DnaK sequences of M. capricolum. Comparison of their dnaK gene sequences with M. capricolum showed 99.64% sequence identity for strain 700 and 99.27% identity for strains ULB-A and ULB-B. In the flock from which M. capricolum-like strains ULB-A and ULB-B were isolated, the majority of chickens (83% of the chickens examined) raised antibodies reacting with M. capricolum antigens. Notably, the infection of chickens with M. capricolum-like strains represents an unusual exception to the range of Mycoplasma species host specificity.

  3. Antibiotic Resistance in Staphylococcus aureus Strains Isolated from Clinical Specimens

    Directory of Open Access Journals (Sweden)

    M. Sirin

    2016-04-01

    Full Text Available Aim: The aim of this study was to determine the antibiotic susceptibilities of S.aureus strains isolated from various clinical specimens between the years 2011-2014 and to investigate the changes of these susceptibilities over the years. Material and Method: Identification and antibiotic susceptibility testing of the strains were performed by Vitek 2 compact automated system (bioMérieux, France. The strains found to be intermediate susceptible to vancomycin and teicoplanin were also tested by E-test method. Results: S.aureus strains (n=1442 were most commonly isolated from wound, urine and blood samples. The isolation rates of methicillin-resistant S.aureus (MRSA in hospitalized patients were significantly higher than the isolation rates of MRSA in outpatients. All strains were susceptible to vancomycin, teicoplanin, linezolid and tigecycline. The total of four years resistance rates of MRSA strains to erythromycin, clindamycin, ciprofloxacin, moxifloxacin, gentamicin, co-trimoxazole, fusidic acid were significantly higher than the resistance rates of methicillin-sensitive S.aureus (MSSA. The changes in the rates of antibiotic resistance were not statistically significant in MSSA strains over the years, and statistically significant decrease was found in erythromycin, clindamycin, ciprofloxacin, moxifloxacin and gentamicin resistance in MRSA strains. Discussion: Glycopeptides, linezolid and tigecycline were the most effective antibiotics against S.aureus strains. It was considered as necessary to detect antimicrobial resistance profiles by effective surveillance studies and monitor the changes occurred over the years in order to prevent the development of resistance and control of infections.

  4. [Determination of in vitro susceptibilities of Brucella spp. strains against 11 different antibacterial gents isolated from blood cultures].

    Science.gov (United States)

    Keşli, Recep; Bilgin, Hüseyin; Yılmaz, Halim

    2017-07-01

    Brucellosis is a worldwide zoonotic disease and still continuous to be a major public health problem. In this study, it was aimed to identify the Brucella strains to the species level isolated from blood cultures, and to determine the rate of antimicrobial susceptibility against eleven antibacterial agents. A total of 106 Brucella spp. strains were included in the study, which were isolated from blood cultures in University of Health Sciences, Konya Training and Research Hospital, Medical Microbiology Laboratory between January 2011 and June 2013. Identification of the isolated strains were mainly based on conventional methods. In vitro antibacterial susceptibilities of azithromycin, ciprofloxacin, doxycycline, gentamicin, levofloxacin, moxifloxacin, rifampicin, streptomycin, tetracycline, tigecycline, and trimethoprim/sulfamethoxazole, were evaluated by using the gradient (E-test, bioMerieux, France) strip method. The bacterial suspensions adjusted to 0.5 McFarland turbidity was inoculated to Mueller Hinton agar plates, supplemented with 5% sheep blood, and E-test strips of selected antibacterial were applied. The plates were incubated in ambient air 48 hours at 37ºC and Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213 were used as quality control strains for antimicrobial susceptibility testing. Minimum inhibitors concentration (MIC) values were interpreted according to Clinical and Laboratory Standards Institute (CLSI) guidelines for slow-growing bacteria such as Haemophilus spp. Of the 106 Brucella spp. strains included in to the study, 90 were identified as Brucella melitensis, and 16 were Brucella abortus. MIC90 values of azithromycin, ciprofloxacin, doxycycline, gentamicin, levofloxacin, moxifloxacin, rifampicin, streptomycin, tetracycline, tigecycline, and trimethoprim/sulfamethoxazole were determined as 1 µg/ml, 0.25 µg/ml, 0.19 µg/ml, 0.25 µg/ml, 0.19 µg/ml, 0.75 µg/ml, 0.25 µg/ml, 0.75 µg/ml, 0.38 µg/ml, 0.64 µg/ml, and 0

  5. [Isolation, identification and genetic analysis of a murine norovirus strain].

    Science.gov (United States)

    Yuan, Wen; Zhang, Yu; Wang, Jing; Liu, Xiang-Mei; Zhao, Wei-Bo; Huang, Ren

    2014-07-01

    Murine norovirus (MNV) was first discovered in mice in 2003. MNV is a member of the genus Norovirus in the family Caliciviridae. It is one of the most important and prevalent pathogens of laboratory mice, and almost all mouse strains are susceptible to MNV infection. In this study, a MNV strain was isolated from the cecal contents of infected mice and identified by the cytopathic effect (CPE) assay, virus plaque assay, 50% tissue culture infectious dose (TCID50) assay, electron microscopy, indirect immunofluorescence assay (IFA) and nucleotide sequencing. On infection, the RAW264.7 cell line showed obvious cytopathic effects within 24 to 48 hours post-inoculation, as infected cells became rounded, bright and shrunken, with ultimate disintegration of the cell sheet. After the isolation of the MNV virus, the virus was plaque-purified in RAW264.7 cells. The TCID50 of the virus was 10(5.25/0.1 mL. Electron microscopic observations of the purified virus showed the presence of spherical and non-enveloped viral particles that were 30 to 35 nm in diameter. According to the identification results, the isolate was named as MNV Guangzhou/K162/09/CHN. Thereafter, five overlapping gene fragments that covered the entire open reading frame (ORF) were amplified by RT-PCR, and the 3'-untranslated region (UTR) and 5'-UTR were amplified using the 3'-rapid amplification of cDNA ends (RACE) and the 5'-RACE method, respectively. Each of the gene fragments were cloned and sequenced, and whole genome sequences of the strain were obtained by assembling the cDNA fragment sequences. The results showed that the length of the complete genome was 7 380 nucleotides (GenBank accession number: HQ317203). The comparison of nucleotide and deduced amino acid sequences of the isolate was performed against other MNV strains in the GenBank database. A phylogenetic tree based on VP1 nucleotide sequences was constructed using MEGA5.0 software. The homology of nucleotides between the MNV Guangzhou/K162

  6. Differential Stimulatory Activities of Smooth and Rough Brucella abortus Lipopolysaccharide in Murine Macrophages

    Directory of Open Access Journals (Sweden)

    Raheela Akhtar1,2*, Yongqun O. He2, Charles B. Larson2, Zafar I. Chaudhary3 and Mansur ud-Din Ahmad4

    2012-06-01

    Full Text Available Brucella abortus lipopolysaccharide (LPS was isolated and purified from rough (RB51 and smooth (S2308 strains of Brucella. The LPS preparations were used to treat murine (RAW 264.7 macrophages in order to study their differential effects. Treated macrophages were tested by lysozyme release test (LRT, nitroblue tetrazolium test (NBT and nitric oxide (NO assay, respectively. Rough Brucella LPS induced significantly higher levels of lysozyme release, oxidative stress, and nitric oxide in murine macrophages than smooth Brucella LPS or combined LPS (rough + smooth LPS. These responses were dose-dependent. Macrophages treated with rough LPS were more Brucellacidal than those treated with smooth LPS. The minimal stimulation of murine macrophages by Brucella smooth LPS may provide basis for less active immune responses against smooth strains.

  7. Proteogenomic Investigation of Strain Variation in Clinical Mycobacterium tuberculosis Isolates

    KAUST Repository

    Heunis, Tiaan

    2017-08-18

    Mycobacterium tuberculosis consists of a large number of different strains that display unique virulence characteristics. Whole-genome sequencing has revealed substantial genetic diversity among clinical M. tuberculosis isolates, and elucidating the phenotypic variation encoded by this genetic diversity will be of utmost importance to fully understand M. tuberculosis biology and pathogenicity. In this study we integrated whole-genome sequencing and mass spectrometry (GeLC-MS/MS) to reveal strain-specific characteristics in the proteomes of two clinical M. tuberculosis Latin American-Mediterranean isolates. Using this approach we identified 59 peptides containing single amino acid variants, which covered ~9% of all total coding nonsynonymous single nucleotide variants detected by whole-genome sequencing. Furthermore, we identified 29 distinct peptides that mapped to a hypothetical protein not present in the M. tuberculosis H37Rv reference proteome. Here we provide evidence for the expression of this protein in the clinical M. tuberculosis SAWC3651 isolate. The strain-specific databases enabled confirmation of genomic differences (i.e. large genomic regions of difference and nonsynonymous single nucleotide variants) in these two clinical M. tuberculosis isolates and allowed strain differentiation at the proteome level. Our results contribute to the growing field of clinical microbial proteogenomics and can improve our understanding of phenotypic variation in clinical M. tuberculosis isolates.

  8. NEW PRODUCER STRAINS OF BIOBUTANOL. І. ISOLATION AND IDENTIFICATION

    Directory of Open Access Journals (Sweden)

    O. A. Tigunova

    2013-02-01

    Full Text Available Getting new, more productive strains of microorganisms that produce butanol is a topical problem. Studing of morphological and physiological characteristics of the isolated strains, improvement of their cultivation conditions, optimization of biobutanol synthesis gives the possibility to organize a cost-effective butanol production technology. The aim of this work was searching new butanol and butyric acid producer strains, their identification and studying the main steps of the selective strains biosynthesis. The objects of this study were microorganisms that had allocated from soils and sludges samples of Kiev’s lakes. Obtained cultures have been screened. Three strains were obtained as promising and identified as C. acetobutylicum, C. tyrobutylicum, C. butylicum. Selective medium have been developed and modified for the microorganisms. Producer’s features were investigated in order to maximize the accumulation of target metabolites.

  9. Characterization of Pasteurella multocida strains isolated from geese.

    Science.gov (United States)

    Varga, Zsuzsanna; Volokhov, Dmitriy V; Stipkovits, László; Thuma, Akos; Sellyei, Boglárka; Magyar, Tibor

    2013-04-12

    Phenotypic and genotypic diversity of forty-two Pasteurella multocida isolates from geese were characterized by analysis of their capsular type, Heddleston serotype, biotype, fimbrial gene allele type, comparative outer membrane protein (OMP) electrophoresis patterns, and were analyzed using PCR for the presence of virulence-associated genes (toxA, tbpA, pfhA, hgbA, hgbB, nanH, nanB, fimA, hsf-1, and pmHAS). A sequence comparison of the thdF and rpoB housekeeping genes of twenty representative P. multocida strains from three different OMP groups demonstrated that seventeen strains were closely related phylogenetically to previously published strains of P. multocida subsp. multocida and P. multocida subsp. gallicida, and only three strains from geese were grouped with previously published strains of P. multocida subsp. septica. This study is the first report regarding the characterization of phenotypic and genotypic features of different P. multocida field strains isolated from geese with the different clinical representation of pasteurellosis and provide our overview on the usefulness of these in vitro tests for primary characterization of P. multocida strains for the epidemiological studies of fowl cholera. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Molecular characterisation of Xanthomonas strains isolated from aroids in Mauritius.

    Science.gov (United States)

    Khoodoo, M H R; Sahin, F; Donmez, M F; Fakim, Y Jaufeerally

    2005-06-01

    Mauritius is one of the largest world producers of Anthurium cut flowers but outbreaks of bacterial blight have never been reported on the island. This work was about the characterisation and identification of bacterial strains isolated from Anthurium andreanum, Dieffenbachia maculata and Aglaonema simplex in Mauritius. Fifteen strains, that showed the morphological properties of Xanthomonas on conventional media, were tested on two semi-selective media (Esculin-trehalose and cellobiose-starch). ELISA tests using a panel of monoclonal antibodies were carried out and three out of 15 strains reacted with a Xanthomonas-specific monoclonal antibody (MAb XII). Analysis using four sets of ribosomal primers revealed that the same three Mauritius strains shared conserved PCR products with reference xanthomonads including virulent strains of Xanthomonas axonopodis pv. dieffenbachiae (Xad). BIOLOG tests and the Sherlock Microbial Identification system (MIDI) identified these three new strains at the species level as X. axonopodis. The complementary tests that were carried out clearly confirmed that the three strains are xanthomonads and, moreover, a DNA probe which showed specificity to Xad strains suggested that the three Mauritius strains are non-virulent forms of the pathogen causing Anthurium blight.

  11. Antimicrobial resistance of bacterial strains isolated from avian cellulitis

    Directory of Open Access Journals (Sweden)

    MM Santos

    2014-03-01

    Full Text Available Avian cellulitis is an inflammatory process in the subcutaneous tissue, mainly located in the abdomen and thighs. This problem is commonly observed in poultry at slaughter and it is considered one of the major causes of condemnation of carcasses in Brazil. The aim of this study was to perform the microbial isolation of lesions of avian cellulitis from a processing plant located in the State of Goiás in order to analyze antimicrobial resistance by antibiogram test and to detect resistance genes by polymerase chain reaction. A total of 25 samples of avian cellulitis lesions were analyzed, from which 30 bacterial strains were isolated. There were eleven (44% strains of Escherichia coli, nine (36% strains of Staphylococcus epidermidis, seven (28% strains of Proteus mirabilis and three (12% strains of Manheimiahaemolytica. The antibiogram test showed that all strains were resistant to at least one antimicrobial. The gene of antimicrobial resistance tetB was detected in E. coli, S. epidermidis and P. mirabilis strains, and was the most frequently observed gene. The gene of antimicrobial resistance Sul1 was detected in all bacterial species, while tetA was found in E. coli and S. epidermidis strains, SHV in E. coli strains, S. epidermidis and P. mirabilis,and cat1 in one P. mirabilis strain. The results suggest a potential public health hazard due to the ability of these microorganisms to transmit antimicrobial resistancegenes to other microorganisms present in the intestinal tract of humans and animals, which may affect clinical-medical usage of these drugs.

  12. Characteristics of an Aeromonas trota strain isolated from cerebrospinal fluid.

    Science.gov (United States)

    Dallagassa, Cibelle B; Surek, Monica; Vizzotto, Bruno S; Prediger, Karoline C; Moriel, Bárbara; Wolf, Suélen; Weiss, Vinícius; Cruz, Leonardo M; Assis, Flávia E A; Paludo, Katia S; Rego, Fabiane G M; Farah, Sônia M S S; Picheth, Geraldo; Souza, Emanuel M; Pedrosa, Fábio O; Chubatsu, Leda S; Fadel-Picheth, Cyntia M T

    2018-01-16

    Aeromonas are ubiquitous in aquatic habitats. However some species can cause infections in humans, but rarely meningitis. Here we describe the isolation and characterization of an Aeromonas strain from cerebrospinal fluid of a meningitis patient. The isolate, identified as A. trota by biochemical and molecular methods, was susceptible to ampicillin but resistant to cephalothin and cefazolin. Genome sequencing revealed virulence factor genes such as type VI secretion system, aerolysin and lateral flagella. The isolate exhibited swarming motility, hemolytic activity and adhesion and cytotoxicity on HeLa cells. This is the first report of A. trota associated with meningitis and its virulence characteristics. Copyright © 2018. Published by Elsevier Ltd.

  13. Bioactivity characterization of Lactobacillus strains isolated from dairy products.

    Science.gov (United States)

    Haghshenas, Babak; Nami, Yousef; Haghshenas, Minoo; Abdullah, Norhafizah; Rosli, Rozita; Radiah, Dayang; Khosroushahi, Ahmad Yari

    2015-10-01

    This study aimed to find candidate strains of Lactobacillus isolated from sheep dairy products (yogurt and ewe colostrum) with probiotic and anticancer activity. A total of 100 samples were randomly collected from yogurt and colostrum and 125 lactic acid bacteria were isolated. Of these, 17 Lactobacillus strains belonging to five species (L. delbrueckii, L. plantarum, L. rhamnosus, L. paracasei, and L. casei) were identified. L. plantarum 17C and 13C, which isolated from colostrums, demonstrated remarkable results such as resistant to low pH and high concentrations of bile salts, susceptible to some antibiotics and good antimicrobial activity that candidate them as potential probiotics. Seven strains (1C, 5C, 12C, 13C, 17C, 7M, and 40M), the most resistant to simulated digestion, were further investigated to evaluate their capability to adhere to human intestinal Caco-2 cells. L. plantarum 17C was the most adherent strain. The bioactivity assessment of L. plantarum 17C showed anticancer effects via the induction of apoptosis on HT-29 human cancer cells and negligible side effects on one human epithelial normal cell line (FHs 74). The metabolites produced by this strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  14. Cephalosporin resistant bacterial strains isolated from respiratory infections

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    Botnarciuc Mihaela

    2015-02-01

    Full Text Available Objectives: The objective of the study is the evaluation of the actual resistance to second, third, and fourth generation cephalosporins over bacterial strains isolated from respiratory infections. The main causes for cephalosporin resistance of pathogenic and conditioned pathogen bacteria are: widespread usage, and impair immune response.

  15. Typing of Staphylococcus aureus strains isolated from milk cows ...

    African Journals Online (AJOL)

    Surveys conducted in Senegal have shown a strong association of staphylococci with subclinical mastitis in dairy cows. This study aimed to characterise Staphylococcus aureus strains identified in the dairy farms in Dakar. Of a total of 244 Staphylococcus spp isolates col ected from 135 lactating cows with subclinical ...

  16. Characterization and Strains of Mycobacteria Isolated from Cattle ...

    African Journals Online (AJOL)

    This work is a pilot study looking kind conducted from March to October 2011 and to determine the frequency of M. bovis strains isolated from cattle carcasses in ... The study also helped to establish the 16 samples positive culture of Mycobacterium bovis distribution of M. bovis based on race, sex, age of cattle and make ...

  17. Investigation of possible virulence factors in Candida strains isolated ...

    African Journals Online (AJOL)

    Background: The Candida species, which are one of the most common causes of nosocomial bloodstream infections, present with high mortality and morbidity rates. This study aims to investigate the production of esterase, phospholipase, proteinase, and biofilm formation ability of the Candida strains isolated from the ...

  18. Probiotic characteristics of Lactobacillus fermentum strains isolated from tulum cheese.

    Science.gov (United States)

    Tulumoğlu, Şener; Kaya, Halil İbrahim; Şimşek, Ömer

    2014-12-01

    The aim of this study was to characterize the probiotic characteristics of Lactobacillus fermentum strains isolated from Tulum cheese. Seven L. fermentum strains were selected among the isolated and identified lactobacillus strains due to their abundance. When the gastric condition was considered, L. fermentum LP3 and LP4 were able to tolerate pH 2.5 and 1% bile salt. All L. fermentum strains had similar enzymatic activity and antibiotic resistance pattern but the highest antagonistic effect was detected within LP3, LP4 and LP6. Cholesterol assimilation amount of L. fermentum strains ranged between 12.1 and 45.3% in MRS and 20.7-71.1% in MRS with bile. The highest cholesterol assimilation in MRS and MRS with bile was occurred by LP3 and LP4, respectively. L. fermentum LP2 adhered to caco-2 cells more than Lactobacillus rhamnosus LGG where LP3, LP4 and LP5 adhered at similar level. In conclusion, L. fermentum LP3 and LP4 fulfilled sufficient criteria to be probiotics for use as a starter culture in the production of tulum cheese or other dairy products. Also this study indicated that some food-associated Lactobacillus strains non-predominant for gut biota have significant probiotic potential. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Pathogenic Potential of Saccharomyces Strains Isolated from Dietary Supplements

    Science.gov (United States)

    Monteoliva, Lucía; Querol, Amparo; Molina, María; Fernández-Espinar, María T.

    2014-01-01

    Saccharomyces cerevisiae plays a beneficial role in health because of its intrinsic nutritional value and bio-functional properties, which is why it is also used as a dietary supplement. However, the perception that S. cerevisiae is harmless has changed due to an increasing number of infections caused by this yeast. Given this scenario, we have tested whether viable strains contained in dietary supplements displayed virulence-associated phenotypic traits that could contribute to virulence in humans. We have also performed an in vivo study of the pathogenic potential of these strains using a murine model of systemic infection by intravenous inoculation. A total of 5 strains were isolated from 22 commercial products and tested. Results highlight one strain (D14) in terms of burden levels in brains and kidneys and ability to cause death, whereas the other two strains (D2 and D4) were considered of low virulence. Our results suggest a strong relationship between some of the virulence-associated phenotypic traits (ability to grow at 39°C and pseudohyphal growth) and the in vivo virulence in a mouse model of intravenous inoculation for isolates under study. The isolate displaying greatest virulence (D14) was evaluated in an experimental murine model of gastrointestinal infection with immunosuppression and disruption of mucosal integrity, which are common risk factors for developing infection in humans, and results were compared with an avirulent strain (D23). We showed that D14 was able to spread to mesenteric nodes and distant organs under these conditions. Given the widespread consumption of dietary supplements, we recommend only safe strains be used. PMID:24879417

  20. Genome Sequence of Campylobacter jejuni strain 327, a strain isolated from a turkey slaughterhouse

    DEFF Research Database (Denmark)

    Takamiya, Monica; Özen, Asli Ismihan; Rasmussen, Morten

    2011-01-01

    , catalase positive bacterium obtains energy from the metabolism of amino acids and Krebs cycle intermediates. Strain 327 was isolated from a turkey slaughter production line and is considered environmentally sensitive to food processing (cold, heat, drying) and storage conditions. The 327 whole genome...... shotgun sequence of 1,618,613 bp long consists of 1,740 protein-coding genes, 46 tRNA genes and 3 rRNA operons. A protein based BLAST analysis places the turkey isolate 327 close to the human clinical strain 81116 (NCTC 11828)....

  1. A novel strain D5 isolated from Acacia confusa.

    Science.gov (United States)

    Huang, Baoling; Lv, Chengqun; Zhao, Yili; Huang, Rong

    2012-01-01

    We isolated a novel strain D5 from nodules of Acacia confusa. Under strict sterile conditions the strain could successfully nodulate Acacia confusa, A. crassicarpa and A. mangium, with nitrogenase activity ranging from 18.90 to 19.86 nmol·g(-1)·min(-1). In the phylogenetic tree based on a complete 16S rRNA gene sequence, the sequence of strain D5 shared 99% homology with that of four species of genus Pseudomonas. The 685 bp nodA fragment amplified from strain D5 shared 95% homology with the nodA sequence of 9 species of genus Bradyrhizobium, with a genetic distance of 0.01682. The 740 bp nifH gene fragment was amplified from strain D5. This strain D5 nifH gene and Bradyrhizobium spp. formed a branch, showing 98% homology and a genetic distance of 0. The homology between this branch and the Bradyrhizobium spp. DG in another branch was 99%, with a genetic distance of 0.007906. These results indicate that this strain D5 is a new type of nitrogen-fixing bacterium.

  2. Development of an in vivo model of Chlamydia abortus chronic infection in mice overexpressing IL-10.

    Science.gov (United States)

    Del Río, Laura; Murcia, Antonio; Buendía, Antonio J; Álvarez, Daniel; Ortega, Nieves; Navarro, José A; Salinas, Jesús; Caro, María Rosa

    2018-01-01

    Chlamydia abortus, like other members of the family Chlamydiaceae, have a unique intracellular developmental cycle that is characterized by its chronic nature. Infection of a flock can remain undetected for months, until abortion occurs the following reproductive season but, to date, neither the location nor the mechanisms that maintain this latent phase are fully understood. Studies have shown that IL-10 produced as a response to certain micro-organisms sustains the intracellular survival of pathogens and increases host susceptibility to chlamydial infections. In order to induce a sustained infection C. abortus, transgenic mice that constitutively express IL-10 were infected and the immunological mechanisms that maintain infection in these mice were compared with the mechanisms of a resistant wild-type mouse strain. Viable bacteria could be detected in different tissues of transgenic mice up to 28 days after infection, as analysed by bacterial isolation and immunohistochemistry. Chronic infection in these mice was associated with an impaired recruitment of macrophages, decreased iNOS activity at the site of infection and a more diffuse distribution of inflammatory cells in the liver. This murine model can be of great help for understanding the immunological and bacterial mechanisms that lead to chronic chlamydial infections. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. An analytical model of strain isolation for stretchable and flexible electronics

    Science.gov (United States)

    Cheng, H.; Wu, J.; Li, M.; Kim, D.-H.; Kim, Y.-S.; Huang, Y.; Kang, Z.; Hwang, K. C.; Rogers, J. A.

    2011-02-01

    One important aspect of stretchable electronics design is to shield the active devices from strains through insertion of a soft layer between devices and substrate. An analytical model is established, which gives linear dependence of strain isolation on the reciprocal of strain-isolation layer thickness, and the reciprocal of device and substrate stiffness. Strain isolation is also linearly proportional to the shear modulus of strain-isolation layer and square of device length.

  4. Multivalent Fusion DNA Vaccine against Brucella abortus

    Science.gov (United States)

    Gómez, Leonardo; Llanos, Javiera; Escalona, Emilia; Sáez, Darwin; Álvarez, Francisco; Molina, Raúl; Flores, Manuel

    2017-01-01

    As an alternative brucellosis prevention method, we evaluated the immunogenicity induced by new multivalent DNA vaccines in BALB/c mice. We constructed the vaccines by fusion of BAB1_0273 and/or BAB1_0278 open reading frames (ORFs) from genomic island 3 (GI-3) and the Brucella abortus 2308 sodC gene with a link based on prolines and alanines (pV273-sod, pV278-sod, and pV273-278-sod, resp.). Results show that immunization with all tested multivalent DNA vaccines induced a specific humoral and cellular immune response. These novel multivalent vaccines significantly increased the production of IgM, IgG, and IgG2a antibodies as well as IFN-γ levels and the lymphoproliferative response of splenocytes. Although immunization with these multivalent vaccines induced a typical T-helper 1- (Th1-) dominated immune response, such immunogenicity conferred low protection levels in mice challenged with the B. abortus 2308 pathogenic strain. Our results demonstrated that the expression of BAB1_0273 and/or BABl_0278 antigens conjugated to SOD protein can polarize mice immunity to a Th1-type phenotype, conferring low levels of protection. PMID:29082252

  5. Multivalent Fusion DNA Vaccine against Brucella abortus

    Directory of Open Access Journals (Sweden)

    Leonardo Gómez

    2017-01-01

    Full Text Available As an alternative brucellosis prevention method, we evaluated the immunogenicity induced by new multivalent DNA vaccines in BALB/c mice. We constructed the vaccines by fusion of BAB1_0273 and/or BAB1_0278 open reading frames (ORFs from genomic island 3 (GI-3 and the Brucella abortus 2308 sodC gene with a link based on prolines and alanines (pV273-sod, pV278-sod, and pV273-278-sod, resp.. Results show that immunization with all tested multivalent DNA vaccines induced a specific humoral and cellular immune response. These novel multivalent vaccines significantly increased the production of IgM, IgG, and IgG2a antibodies as well as IFN-γ levels and the lymphoproliferative response of splenocytes. Although immunization with these multivalent vaccines induced a typical T-helper 1- (Th1- dominated immune response, such immunogenicity conferred low protection levels in mice challenged with the B. abortus 2308 pathogenic strain. Our results demonstrated that the expression of BAB1_0273 and/or BABl_0278 antigens conjugated to SOD protein can polarize mice immunity to a Th1-type phenotype, conferring low levels of protection.

  6. Pathogenicity of rabies viruses isolated in China: two fixed strains and a street strain.

    Science.gov (United States)

    Huang, Ying; Tang, Qing; Rayner, Simon; Gong, Kai; Song, Bo; Liang, Guo Dong

    2013-07-01

    To investigate the virulence characteristics of two fixed strains (CTN and aG) and a street strain (HN10) of rabies viruses isolated in China. ICR mice of different age groups were inoculated with CTN, aG and HN10 rabies virus strains via the intracracerebral (i.c.) or intramuscular (i.m.) routes, and observed for 20 days. The CTN strain was pathogenic to 7-day-old suckling mice that received i.c. inoculations and 3-day-old suckling mice that received i.m. inoculations. The aG strain was pathogenic to 4-week-old mice that received i.c. inoculations and 7-day-old suckling mice that received i.m. inoculations. The HN10 strain was pathogenic to mice of all age groups via both inoculation routes. In moribund mice, the viruses had spread to most regions of the brain. The CTN and HN10 strains had similar dissemination patterns in the brain; both viral antigens could be found in the dentate gyrus (DG), whereas few viral antigens were present in the DG from specimens that had been infected with the aG strain. A comprehensive sequence analysis of the G protein suggested that differences in gene sequences may be responsible for producing strain-specific differences in pathogenicity and distribution in the brain. Copyright © 2013 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  7. Evaluation of cell-mediated immune responses and bacterial clearance in 6-10 months old water buffalo (Bubalus bubalis) experimentally vaccinated with four dosages of commercial Brucella abortus strain RB51 vaccine.

    Science.gov (United States)

    Diptee, M D; Adesiyun, A A; Asgarali, Z; Campbell, M; Fosgate, G T

    2005-07-15

    Thirty water buffalo, obtained from a brucellosis-free farm, were used to evaluate cell-mediated immune responses and bacterial clearance in response to vaccination with Brucella abortus strain RB51 (RB51) in a dose-response study. The animals were randomly divided into five treatment groups. Groups I--V received the recommended dose (RD) of RB51 vaccine once, RD twice 4 weeks apart, double RD once, double RD twice 4 weeks apart and saline once, respectively. Cell-mediated immune response to RB51 was assessed by the histological examination of haematoxylin and eosin (H&E) stained sections of lymph nodes draining the sites of inoculation and by comparison of stimulation indices (SI) derived from gamma interferon (IFN-gamma) assay. A mixture of cytoplasmic proteins from B. melitensis B115 (brucellergene) was used as a specific antigenic stimulus to peripheral blood mononuclear cells (PBMC) and lymph node mononuclear cells (LNMC) up to 22 post-initial-inoculation week (PIW). Supernatants harvested at 18-24h after the in vitro antigenic stimulus were assayed for their IFN-gamma content by using a commercial sandwich enzyme-linked immunosorbent assay (ELISA) kit. Clearance of RB51 was assessed by the sequential immunohistochemical examination of sections of draining lymph nodes post-inoculation. There was no observable expansion of the deep cortex of lymph nodes on H&E sections indicating poor T-cell stimulation. All group V (control) water buffalo PBMC ELISA values were negative (SIRB51 occurred between 4 and 6 PIW in treatment groups I and III and between 6 and 12 PIW in groups II and IV. RB51 was not detected in any of the control animals at sampling intervals post-inoculation.

  8. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Purification of Helicobacter suis Strains From Biphasic Cultures by Single Colony Isolation: Influence on Strain Characteristics.

    Science.gov (United States)

    Liang, Jungang; De Bruyne, Ellen; Ducatelle, Richard; Smet, Annemieke; Haesebrouck, Freddy; Flahou, Bram

    2015-06-01

    Helicobacter (H.) suis causes gastritis and decreased weight gain in pigs. It is also the most prevalent non-Helicobacter pylori Helicobacter species in humans with gastric disease. H. suis is extremely fastidious, and so far, biphasic culture conditions were essential for isolation and culture, making it impossible to obtain single colonies. Hence, cultures obtained from an individual animal may contain multiple H. suis strains, which is undesirable for experiments aiming for instance at investigating H. suis strain differences. Pure cultures of H. suis were established by growing bacteria as colonies on 1% brucella agar plates, followed by purification and enrichment by biphasic subculture. Characteristics of these single colony-derived strains were compared with those of their parent strains using multilocus sequence typing (MLST) and by studying bacterium-host interactions using a gastric epithelial cell line and Mongolian gerbil model. The purification/enrichment procedure required a nonstop culture of several weeks. For 4 of 17 H. suis strains, MLST revealed differences between parental and single colony-derived strains. For three of four single colony-derived strains tested, the cell death-inducing capacity was higher than for the parental strain. One single colony-derived strain lost its capacity to colonize Mongolian gerbils. For the four other strains tested, colonization capacity and histopathologic changes were similar to what has been described when using strains with only a history of limited biphasic culture. A method was developed to obtain single colony-derived H. suis strains, but this procedure may affect the bacterial genotype and phenotype. © 2015 John Wiley & Sons Ltd.

  10. Characterization of Genomic Island 3 and Genetic Variability of Chilean Field Strains of Brucella abortus▿

    Science.gov (United States)

    Céspedes, Sandra; Salgado, Paulina; Valenzuela, Patricio; Vidal, Roberto; Oñate, Angel A.

    2011-01-01

    One of the capabilities developed by bacteria is the ability to gain large fragments of DNA from other bacteria or to lose portions of their own genomes. Among these exchangeable fragments are the genomic islands (GIs). Nine GIs have been identified in Brucella, and genomic island 3 (GI-3) is shared by two pathogenic species, B. melitensis and B. abortus. GI-3 encodes mostly unknown proteins. One of the aims of this study was to perform pulsed-field gel electrophoresis (PFGE) on field isolates of B. abortus from Chile to determine whether these isolates are clonally related. Furthermore, we focused on the characterization of GI-3, studying its organization and the genetic conservation of the GI-3 sequence using techniques such as tiling-path PCR (TP-PCR) and restriction fragment length polymorphism-PCR (RFLP-PCR). Our results, after PFGE was performed on 69 field isolates of B. abortus from Chile, showed that the strains were genetically homogeneous. To increase the power of genetic discrimination among these strains, we used multiple locus variable-number tandem-repeat (VNTR) analysis with 16 loci (MLVA-16). The results obtained by MLVA-16 showed that the strains of B. abortus were genetically heterogeneous and that most of them clustered according to their geographic origin. Of the genetic loci studied, panel 2B was the one describing the highest diversity in the analysis, as well as locus Bruce19 in panel 2A. In relation to the study of GI-3, our experimental analysis by TP-PCR identified and confirmed that GI-3 is present in all wild strains of B. abortus, demonstrating the high stability of gene cluster GI-3 in Chilean field strains. PMID:21543580

  11. Caspase-2 mediated apoptotic and necrotic murine macrophage cell death induced by rough Brucella abortus.

    Directory of Open Access Journals (Sweden)

    Fang Chen

    Full Text Available Brucella species are Gram-negative, facultative intracellular bacteria that cause zoonotic brucellosis. Survival and replication inside macrophages is critical for establishment of chronic Brucella infection. Virulent smooth B. abortus strain 2308 inhibits programmed macrophage cell death and replicates inside macrophages. Cattle B. abortus vaccine strain RB51 is an attenuated rough, lipopolysaccharide O antigen-deficient mutant derived from smooth strain 2308. B. abortus rough mutant RA1 contains a single wboA gene mutation in strain 2308. Our studies demonstrated that live RB51 and RA1, but not strain 2308 or heat-killed Brucella, induced both apoptotic and necrotic cell death in murine RAW264.7 macrophages and bone marrow derived macrophages. The same phenomenon was also observed in primary mouse peritoneal macrophages from mice immunized intraperitoneally with vaccine strain RB51 using the same dose as regularly performed in protection studies. Programmed macrophage cell death induced by RB51 and RA1 was inhibited by a caspase-2 inhibitor (Z-VDVAD-FMK. Caspase-2 enzyme activation and cleavage were observed at the early infection stage in macrophages infected with RB51 and RA1 but not strain 2308. The inhibition of macrophage cell death promoted the survival of rough Brucella cells inside macrophages. The critical role of caspase-2 in mediating rough B. abortus induced macrophage cell death was confirmed using caspase-2 specific shRNA. The mitochondrial apoptosis pathway was activated in macrophages infected with rough B. abortus as demonstrated by increase in mitochondrial membrane permeability and the release of cytochrome c to cytoplasm in macrophages infected with rough Brucella. These results demonstrate that rough B. abortus strains RB51 and RA1 induce apoptotic and necrotic murine macrophage cell death that is mediated by caspase-2. The biological relevance of Brucella O antigen and caspase-2-mediated macrophage cell death in Brucella

  12. Caspase-2 mediated apoptotic and necrotic murine macrophage cell death induced by rough Brucella abortus.

    Science.gov (United States)

    Chen, Fang; He, Yongqun

    2009-08-28

    Brucella species are Gram-negative, facultative intracellular bacteria that cause zoonotic brucellosis. Survival and replication inside macrophages is critical for establishment of chronic Brucella infection. Virulent smooth B. abortus strain 2308 inhibits programmed macrophage cell death and replicates inside macrophages. Cattle B. abortus vaccine strain RB51 is an attenuated rough, lipopolysaccharide O antigen-deficient mutant derived from smooth strain 2308. B. abortus rough mutant RA1 contains a single wboA gene mutation in strain 2308. Our studies demonstrated that live RB51 and RA1, but not strain 2308 or heat-killed Brucella, induced both apoptotic and necrotic cell death in murine RAW264.7 macrophages and bone marrow derived macrophages. The same phenomenon was also observed in primary mouse peritoneal macrophages from mice immunized intraperitoneally with vaccine strain RB51 using the same dose as regularly performed in protection studies. Programmed macrophage cell death induced by RB51 and RA1 was inhibited by a caspase-2 inhibitor (Z-VDVAD-FMK). Caspase-2 enzyme activation and cleavage were observed at the early infection stage in macrophages infected with RB51 and RA1 but not strain 2308. The inhibition of macrophage cell death promoted the survival of rough Brucella cells inside macrophages. The critical role of caspase-2 in mediating rough B. abortus induced macrophage cell death was confirmed using caspase-2 specific shRNA. The mitochondrial apoptosis pathway was activated in macrophages infected with rough B. abortus as demonstrated by increase in mitochondrial membrane permeability and the release of cytochrome c to cytoplasm in macrophages infected with rough Brucella. These results demonstrate that rough B. abortus strains RB51 and RA1 induce apoptotic and necrotic murine macrophage cell death that is mediated by caspase-2. The biological relevance of Brucella O antigen and caspase-2-mediated macrophage cell death in Brucella pathogenesis and

  13. [Antimicrobial susceptibility of Helicobacter pylori strains isolated from gastric lesions].

    Science.gov (United States)

    Pereyra, Laura Del V; Gorordo Ipiña, Rosa C; Berruezo, Fabiana A; Amieva, Cristian A; García, Maria E; Bottiglieri, Marina T

    Antimicrobial susceptibility was evaluated by two diffusion methods: E-test strips to determine minimum inhibitory concentration (MIC) and disk diffusion for amoxicillin, clarithromycin, metronidazole and ciprofloxacin in 30 Helicobacter pylori strains isolated from gastric biopsies. No strains were resistant to amoxicillin, 17% (5/30) were resistant to clarithromycin, 20% (6/30) ciprofloxacin by both methods, and 37% (11/30) to metronidazole by the E-test. Although the number of strains studied was reduced, there was a single mismatch in interpreting susceptibility when both methods were compared; the same mismatch was observed for metronidazole, being categorized as sensitive by the E-test and as intermediate by disk diffusion. No association between the histological type of lesion and the resistance pattern found could be determined. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  14. [Frequency of isolation and susceptibility to antibiotics of Escherichia coli strains isolated from blood].

    Science.gov (United States)

    Białucha, Agata; Budzyńska, Anna; Kozuszko, Sylwia; Gospodarek, Eugenia

    2010-01-01

    The aim of our study was the analysis of Escherichia coli strains obtained from patients of University Hospital No 1 of dr A. Jurasz Collegium Medicum of L. Rydygier in Bydgoszcz Nicolaus Copernicus University in Toruń and State Infectious Diseases Observatory Hospital of T. Browicz in Bydgoszcz, between 2007 and 2010. Among all microorganisms isolated from blood was 8.7% E. coli strains and 45.1% of all rods from Enterobacteriaceae family. Number of E. coli isolations from positive blood samples was: 64 in 2007, 69 in 2008, 77 in 2009 and 26 in the first half of 2010 year. The highest percentage of E. coli strains were obtained from patients of the Transplantology and Surgery Clinic (16.1%), the Nephrology and Internal Diseases Clinic with the Dialysis Centre (14.0%), the Pediatric, Hematology and Oncology Clinic (13.6%) and the Anesthesiology and Intensive Care Clinic (13.6%). All analysed strains were susceptible to carbapenems, amikacin and tygecycline. The highest percentage of resistant strains were observed to ampicillin (70.7%), piperacillin (43.9%), tetracycline (42.8%) and doxycycline (38.8%). During four years of study 4 (6.3%), one, three and two E. coli strains with ESBL were isolated, respectively.

  15. Probiotic potential of Enterococcus faecalis strains isolated from meconium

    Directory of Open Access Journals (Sweden)

    Ahmed eKhassaf Al Atyaa

    2015-04-01

    Full Text Available 107 bacterial isolates with Gram positive staining and negative catalase activity, presumably assumed as LAB, were isolated from samples of meconium of six donors at Roubaix hospital, in the north of France. All these bacterial isolates were identified by MALDI-TOF mass spectrometry as Enterococcus faecalis. However, only six isolates among which E. faecalis 14, E. faecalis 28, E. faecalis 90, E. faecalis 97 and E. faecalis 101 (obtained from donor 3 and E. faecalis 93 (obtained from donor 5 were active against some Gram negative bacteria (GNB and Gram positive bacteria (GPB, through production of lactic acid and bacteriocin like inhibitory substances (BLIS. The identification of these isolates was confirmed by 16rDNA sequencing and their genetic relatedness was established by REP-PCR and pulsed field gel electrophoresis (PFGE methods. Importantly, the aforementioned antagonistic isolates were sensitive to various classes of antibiotics tested, exhibited high scores of coaggregation and hydrophobicity, and were not hemolytic. Taken together, these properties render these strains as potential candidates for probiotic applications.

  16. Biodegradation of acetochlor by a newly isolated Pseudomonas strain.

    Science.gov (United States)

    Luo, Wei; Gu, Qiuya; Chen, Wenting; Zhu, Xiangcheng; Duan, Zhibing; Yu, Xiaobin

    2015-05-01

    A novel microbial strain JD115 capable of degrading acetochlor was isolated from the sludge of acetochlor manufacture and was identified as Pseudomonas aeruginosa species. This strain was able to grow on acetochlor as the sole source of both carbon and nitrogen. The biodegradation of acetochlor by strain JD115 could be described either by the pseudo-first-order or by the second-order kinetics models, while the latter gave a better performance. The strain optimally degraded acetochlor at a pH value of 7.0 and a temperature of 37 °C. Additional nutriments could greatly enhance the degradation rate of acetochlor up to 95.4% in the presence of 50 mg acetochlor l(-1). The metabolite analyses by GC-MS presumed that catechol was an intermediate product of acetochlor, which was finally degraded for 5 days of incubation. This study highlights the potential use of this strain for the bioremediation of an acetochlor-polluted environment.

  17. Probiotic properties of Lactobacillus strains isolated from Tibetan kefir grains.

    Science.gov (United States)

    Zheng, Yongchen; Lu, Yingli; Wang, Jinfeng; Yang, Longfei; Pan, Chenyu; Huang, Ying

    2013-01-01

    The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB) isolated from Tibetan kefir grains. Three Lactobacillus isolates identified as Lactobacillus acidophilus LA15, Lactobacillus plantarum B23 and Lactobacillus kefiri D17 that showed resistance to acid and bile salts were selected for further evaluation of their probiotic properties. The 3 selected strains expressed high in vitro adherence to Caco-2 cells. They were sensitive to gentamicin, erythromycin and chloramphenicol and resistant to vancomycin with MIC values of 26 µg/ml. All 3 strains showed potential bile salt hydrolase (BSH) activity, cholesterol assimilation and cholesterol co-precipitation ability. Additionally, the potential effect of these strains on plasma cholesterol levels was evaluated in Sprague-Dawley (SD) rats. Rats in 4 treatment groups were fed the following experimental diets for 4 weeks: a high-cholesterol diet, a high-cholesterol diet plus LA15, a high-cholesterol diet plus B23 or a high-cholesterol diet plus D17. The total cholesterol, triglyceride and low-density lipoprotein cholesterol levels in the serum were significantly (P<0.05) decreased in the LAB-treated rats compared with rats fed a high-cholesterol diet without LAB supplementation. The high-density lipoprotein cholesterol levels in groups B23 and D17 were significantly (P<0.05) higher than those in the control and LA15 groups. Additionally, both fecal cholesterol and bile acid levels were significantly (P<0.05) increased after LAB administration. Fecal lactobacilli counts were significantly (P<0.05) higher in the LAB treatment groups than in the control groups. Furthermore, the 3 strains were detected in the rat small intestine, colon and feces during the feeding trial. The bacteria levels remained high even after the LAB administration had been stopped for 2 weeks. These results suggest that these strains may be used in the future as probiotic starter cultures for manufacturing

  18. Isolation, identification and antibiotic resistance of Campylobacter strains isolated from domestic and free-living pigeons.

    Science.gov (United States)

    Dudzic, A; Urban-Chmiel, R; Stępień-Pyśniak, D; Dec, M; Puchalski, A; Wernicki, A

    2016-04-01

    1. The aim of this study was to evaluate the occurrence of Campylobacter spp. in domestic and free-living pigeons and to evaluate the antibiotic resistance profiles. 2. The material consisted of cloacal swabs obtained from 108 homing pigeons and fresh faeces from 72 wild birds from Lublin and its vicinity. The identification of strains isolated on differential/selective media for Campylobacter spp. was carried out by MALDI-TOF and PCR. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in Mueller-Hinton broth. 3. A total of 35 strains of Campylobacter spp. were isolated; 27 were identified as Campylobacter jejuni and 8 as Campylobacter coli. Over half of the isolates were resistant to erythromycin and streptomycin, 40% of strains were resistant to tetracycline and ampicillin and 37% isolates were resistant to amoxicillin. Resistance to two or more antibiotics was observed in all strains tested. 4. The results indicate that both domestic and free-living pigeons are reservoirs for bacteria of the genus Campylobacter, which are characterised by varied and growing resistance to commonly used antibiotics.

  19. [Strains of Klebsiella isolated from patients Łódź hospitals in 2006 year].

    Science.gov (United States)

    Szczerba, Izabela; Gortat, Katarzyna; Majewski, Karol

    2009-01-01

    The aim of this study was to determine a susceptibility of 223 clinical strains Klebsiella to 11 antimicrobial agents and occurrence ESBL-positive strains. These strains were isolated from clinical specimens during 2006 year. Susceptibility of these strains against antimicrobial agents was determined with disc diffusion method according to CLSI recommendation. None of the analysed strains was resistant to imipenem. The Klebsiella pneumonie strains were most frequently isolated from respiratory tract.

  20. Isolation and Identification of L-asparaginase producing Erwinia strains which isolated from Potato Farms

    Directory of Open Access Journals (Sweden)

    Arastoo Badoei-Dalfard

    2016-09-01

    Full Text Available Introduction: L-Asparaginase can be effectively used for the treatment of lymphoblastic leukemia. The rapid growth of cancer cells are needed for L-asparagine abundant storage. L-asparaginase catalyzes the hydrolysis of L-asparagine into L-aspartic acid and ammonia. The purpose of this study was to isolate and identify the L-asparaginase producing Erwinia strains from the potato farms of Jiroft. Materials and methods: Pectolytic Erwinia species isolated from crumbling potato in M9 medium. The desired L-asparaginase producing bacteria were isolated based on the color changes. Biochemical-microbial and the plant pathogenicity tests of these strains were also investigated with potato and geranium. The L-asparaginase production and molecular detection of these Erwinia strains were also investigated. Results: In this study, L-asparaginase producing Erwinia was isolated on the CVP and M9 mediums. The inoculation of Erwinia strains on the potato and geranium plants showed that Er8 and Er11 species have the ability to cause plant pathogenicity. Results showed that the maximum pathogenicity of Er8 and Er11 was observed after 48 and 15 h of inoculation in potato and geranium plants, respectively. 16S rDNA sequencing and phylogenetic analyses exhibited that Er8 and Er11 strains were similar to Erwinia chrysanthemi with 98% homology. Discussion and conclusion: Because of several applications of the Erwinia L-asparaginase in various fields, isolated Erwinia and their L-asparaginase can be suitable for applied utilization.

  1. Entamoeba dispar strains: analysis of polymorphism in Tunisian isolates.

    Science.gov (United States)

    Ayed, Soumaya Ben; Bouratbine, Aida

    2013-01-01

    The ability to detect intra-species polymorphism in Entamoeba histolytica and Entamoeba dispar is an important tool for studying geographic distribution and transmission mechanisms. E. dispar and E. histolytica share the same mechanism for transmission among human hosts, and so after differentiation between these species. We studied the intra-species variation and distribution of E. dispar strains obtained from cyst passers, specifically from African students and Tunisian food handlers. We analyzed the polymorphic region of the chitinase protein gene in 13 individuals infected with E. dispar, of which 9 were from Tunisia and 4 from other African countries. We identified 7 different chitinase patterns in Tunisians while the 4 isolates from other countries each had a distinct pattern. Two of the patterns we found have been reported in studies from Mexico and India, possibly indicating worldwide spread of certain strains. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Isolation of an osmotolerant ale strain of Saccharomyces cerevisiae.

    Science.gov (United States)

    Pironcheva, G

    1998-01-01

    Saccharomyces cerevisiae (ale strain) grown in batch culture to stationary phase was tested for its tolerance to heat (50 degrees C for 5 min), hydrogen peroxide (0.3 M) and salt (growth in 1.5 M sodium chloride/YPD medium). Yeast cells which have been exposed previously to heat shock are more tolerant to hydrogen peroxide and high salt concentrations (1.5 M NaCl) than the controls. Their fermentative activity as judged by glucose consumption and their viability, as judged by cell number and density have higher levels when compared with cells not previously exposed to heat shock. Experimental conditions facilitated the isolation of S. cerevisiae ale strain, which was tolerant to heat, and other agents such as hydrogen peroxide and sodium chloride.

  3. Brucella abortus RB51 and Hot Saline Extract from Brucella ovis as Antigens in a Complement Fixation Test Used To Detect Sheep Vaccinated with Brucella abortus RB51

    OpenAIRE

    Adone, Rosanna; Ciuchini, Franco

    2001-01-01

    The efficacy of Brucella abortus RB51 and hot saline extract (HSE) from Brucella ovis as antigens in complement fixation (CF) tests was comparatively evaluated in detecting immune responses of sheep vaccinated with B. abortus strain RB51. For this study, four 5-month-old sheep were vaccinated subcutaneously with 5 × 109 CFU of RB51, and two sheep received saline. Serum samples collected at different times after vaccination were tested for the presence of antibodies to RB51 by a CF test with R...

  4. Complement Fixation Test To Assess Humoral Immunity in Cattle and Sheep Vaccinated with Brucella abortus RB51

    OpenAIRE

    Adone, Rosanna; Ciuchini, Franco

    1999-01-01

    The live attenuated Brucella abortus strain RB51 is a rifampin-resistant, lipopolysaccharide (LPS) O-chain-deficient mutant of virulent B. abortus 2308. The reduced O-chain content in RB51 prevents this bacterium from inducing antibodies detectable by the conventional serologic tests for bovine brucellosis diagnosis that mainly identify antibodies to LPS. The absence of available serologic tests for RB51 also complicates the diagnosis of possible RB51 infections in...

  5. Use of lactic strains isolated from Algerian ewe's milk in the ...

    African Journals Online (AJOL)

    Fifty three strains of thermophilic and mesophilic lactic acid bacteria were isolated from the ewe's milk. Identification reveals the presence of nineteen strains (36%) of Lactobacillus sp., seventeen strains (32%) of Lactococcus sp., nine strains (17%) of Streptococcus thermophilus and eight strains (15%) of Leuconostoc sp.

  6. Deletion of znuA virulence factor attenuates Brucella abortus and confers protection against wild-type challenge.

    Science.gov (United States)

    Yang, Xinghong; Becker, Todd; Walters, Nancy; Pascual, David W

    2006-07-01

    znuA is known to be an important factor for survival and normal growth under low Zn(2+) concentrations for Escherichia coli, Haemophilus spp., Neisseria gonorrhoeae, and Pasteurella multocida. We hypothesized that the znuA gene present in Brucella melitensis 16 M would be similar to znuA in B. abortus and questioned whether it may also be an important factor for growth and virulence of Brucella abortus. Using the B. melitensis 16 M genome sequence, primers were designed to construct a B. abortus deletion mutant. A znuA knockout mutation in B. abortus 2308 (DeltaznuA) was constructed and found to be lethal in low-Zn(2+) medium. When used to infect macrophages, DeltaznuA B. abortus showed minimal growth. Further study with DeltaznuA B. abortus showed that its virulence in BALB/c mice was attenuated, and most of the bacteria were cleared from the spleen within 8 weeks. Protection studies confirmed the DeltaznuA mutant as a potential live vaccine, since protection against wild-type B. abortus 2308 challenge was as effective as that obtained with the RB51 or S19 vaccine strain.

  7. Infection of cattle in Kenya with Brucella abortus biovar 3 and Brucella melitensis biovar 1 genotypes

    NARCIS (Netherlands)

    Muendo, Esther N.; Mbatha, Peter M.; Macharia, Joseph; Abdoel, Theresia H.; Janszen, Paul V.; Pastoor, Rob; Smits, Henk L.

    2012-01-01

    Brucella melitensis biovar 1 was isolated from bovine milk samples from a herd in central Kenya, and Brucella abortus biovar 3 was isolated from aborted fetus materials and vaginal discharge fluids from cattle in central and eastern provinces of Kenya. All infections including those with B.

  8. Arsenite oxidizing Thiomonas strains isolated from different mining sites

    Science.gov (United States)

    Battaglia-Brunet, F.; Duquesne, K.; Dictor, M. C.; Garrido, F.; Bonnefoy, V.; Baranger, P.; Morin, D.

    2003-04-01

    Arsenic is commonly found in sulfide rocks and ores. This toxic metalloid is transferred to the water phase through acidophilic bio-oxidation of sulfides in mining galleries and waste dumps. Inorganic arsenic As(III) and As(V) are both soluble anions, however As(III) is more mobile and toxic than As(V). Bacteria can participate to the biogeochemical arsenic cycling through As(III) oxidation or As(V) reduction. Mineral selective media, containing As(III) as sole energy source, were used to isolate As(III)-oxidizing bacteria from two disused mining sites. Cheni site (Haute Vienne) was a gold mine, and Carnoules (Gard) was lead-zinc mine. Both sites are highly contaminated with arsenic. Samples of sediments and water from Cheni (pH 6) and Carnoules (pH 3) were used to inoculate mineral selective media whose pH were adjusted to those of the sampling environments. In both cases, organisms belonging to the genus Thiomonas were selected, then isolated. These bacteria oxidize arsenite during their exponential growth phase. The Both bacteria are able to grow, as a pure strains, in autotrophic conditions. The As(III)-oxidase activity of the Carnoules strain was exclusively found in cells cultivated with arsenite, and was associated to the membrane. If they can use As(III) as energetic substrate, Thiomonas-related organisms may play an important role in the biogeochemical cycling of arsenic within mining ecosystems.

  9. 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

    OpenAIRE

    V Leyán; D Pesutic; G Schurig; F Wittwer; P A Contreras; J Kruze

    2005-01-01

    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...

  10. 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

    OpenAIRE

    V Leyán; F Wittwer; P A Contreras; G Schurig

    2006-01-01

    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 ...

  11. Gentamicin resistance among Escherichia coli strains isolated in neonatal sepsis.

    Science.gov (United States)

    Hasvold, J; Bradford, L; Nelson, C; Harrison, C; Attar, M; Stillwell, T

    2013-01-01

    Neonatal sepsis is a significant cause of morbidity and mortality among term and preterm infants. Ampicillin and gentamicin are standard empiric therapy for early onset sepsis. Four cases of neonatal sepsis secondary to Escherichia coli (E. coli) found to be gentamicin resistant occurred within a five week period in one neonatal intensive care unit (NICU). To determine whether these cases could be tied to a single vector of transmission, and to more broadly evaluate the incidence of gentamicin resistant strains of E. coli in the neonatal population at our institution compared to other centers, we reviewed the charts of the four neonates (Infants A through D) and their mothers. The E. coli isolates were sent for Pulse Field Gel Electrophoresis (PFGE) to evaluate for genetic similarity between strains. We also reviewed all positive E. coli cultures from one NICU over a two year period. Infants A and B had genetically indistinguishable strains which matched that of urine and placental cultures of Infant B's mother. Infant C had a genetically distinct organism. Infant D, the identical twin of Infant C, did not have typing performed. Review of all cultures positive for E. coli at our institution showed a 12.9 percent incidence of gentamicin-resistance. A review of other studies showed that rates of resistance vary considerably by institution. We conclude that gentamicin-resistant E. coli is a relatively uncommon cause of neonatal sepsis, but should remain a consideration in patients who deteriorate despite initiation of empiric antibiotics.

  12. Probiotic properties of Lactobacillus strains isolated from Tibetan kefir grains.

    Directory of Open Access Journals (Sweden)

    Yongchen Zheng

    Full Text Available The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB isolated from Tibetan kefir grains. Three Lactobacillus isolates identified as Lactobacillus acidophilus LA15, Lactobacillus plantarum B23 and Lactobacillus kefiri D17 that showed resistance to acid and bile salts were selected for further evaluation of their probiotic properties. The 3 selected strains expressed high in vitro adherence to Caco-2 cells. They were sensitive to gentamicin, erythromycin and chloramphenicol and resistant to vancomycin with MIC values of 26 µg/ml. All 3 strains showed potential bile salt hydrolase (BSH activity, cholesterol assimilation and cholesterol co-precipitation ability. Additionally, the potential effect of these strains on plasma cholesterol levels was evaluated in Sprague-Dawley (SD rats. Rats in 4 treatment groups were fed the following experimental diets for 4 weeks: a high-cholesterol diet, a high-cholesterol diet plus LA15, a high-cholesterol diet plus B23 or a high-cholesterol diet plus D17. The total cholesterol, triglyceride and low-density lipoprotein cholesterol levels in the serum were significantly (P<0.05 decreased in the LAB-treated rats compared with rats fed a high-cholesterol diet without LAB supplementation. The high-density lipoprotein cholesterol levels in groups B23 and D17 were significantly (P<0.05 higher than those in the control and LA15 groups. Additionally, both fecal cholesterol and bile acid levels were significantly (P<0.05 increased after LAB administration. Fecal lactobacilli counts were significantly (P<0.05 higher in the LAB treatment groups than in the control groups. Furthermore, the 3 strains were detected in the rat small intestine, colon and feces during the feeding trial. The bacteria levels remained high even after the LAB administration had been stopped for 2 weeks. These results suggest that these strains may be used in the future as probiotic starter cultures for

  13. 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 organism...

  14. Different distribution patterns of ten virulence genes in Legionella reference strains and strains isolated from environmental water and patients.

    Science.gov (United States)

    Zhan, Xiao-Yong; Hu, Chao-Hui; Zhu, Qing-Yi

    2016-04-01

    Virulence genes are distinct regions of DNA which are present in the genome of pathogenic bacteria and absent in nonpathogenic strains of the same or related species. Virulence genes are frequently associated with bacterial pathogenicity in genus Legionella. In the present study, an assay was performed to detect ten virulence genes, including iraA, iraB, lvrA, lvrB, lvhD, cpxR, cpxA, dotA, icmC and icmD in different pathogenicity islands of 47 Legionella reference strains, 235 environmental strains isolated from water, and 4 clinical strains isolated from the lung tissue of pneumonia patients. The distribution frequencies of these genes in reference or/and environmental L. pneumophila strains were much higher than those in reference non-L. pneumophila or/and environmental non-L. pneumophila strains, respectively. L. pneumophila clinical strains also maintained higher frequencies of these genes compared to four other types of Legionella strains. Distribution frequencies of these genes in reference L. pneumophila strains were similar to those in environmental L. pneumophila strains. In contrast, environmental non-L. pneumophila maintained higher frequencies of these genes compared to those found in reference non-L. pneumophila strains. This study illustrates the association of virulence genes with Legionella pathogenicity and reveals the possible virulence evolution of non-L. pneumophia strains isolated from environmental water.

  15. Efficacy of vaccination strategies against intranasal challenge with Brucella abortus in BALB/c mice.

    Science.gov (United States)

    Surendran, Naveen; Sriranganathan, Nammalwar; Lawler, Heather; Boyle, Stephen M; Hiltbold, Elizabeth M; Heid, Bettina; Zimmerman, Kurt; Witonsky, Sharon G

    2011-03-24

    Brucellosis is a zoonotic disease affecting 500,000 people worldwide annually. Inhalation of aerosol containing a pathogen is one of the major routes of disease transmission in humans. Currently there are no licensed human vaccines available. Brucella abortus strain RB51 is a USDA approved live attenuated vaccine against cattle brucellosis. In a mouse model, strain RB51 over-expressing superoxide dismutase (SOD) administered intraperitoneally (IP) has been shown to be more protective than strain RB51 against an IP challenge with B. abortus pathogenic strain 2308. However, there is lack of information on the ability of these vaccine strains to protect against intranasal challenge. With the long-term goal of developing a protective vaccine for animals and people against respiratory challenge of Brucella spp., we tested a number of different vaccination strategies against intranasal infection with strain 2308. We employed strains RB51 and RB51SOD to assess the efficacy of route, dose, and prime-boost strategies against strain 2308 challenge. Despite using multiple protocols to enhance mucosal and systemic protection, neither rough RB51 vaccine strains provided respiratory protection against intranasal pathogenic Brucella infection. However, intranasal (IN) administration of B. abortus vaccine strain 19 induced significant (p≤0.05) pulmonary clearance of strain 2308 upon IN challenge infection compared to saline. Further studies are necessary to address host-pathogen interaction in the lung microenvironment and elucidate immune mechanisms to enhance protection against aerosol infection. Published by Elsevier Ltd.

  16. Comparison of Mycoplasma pneumoniae genome sequences from strains isolated from symptomatic and asymptomatic patients

    Directory of Open Access Journals (Sweden)

    Emiel B.M. Spuesens

    2016-10-01

    Full Text Available Mycoplasma pneumoniae is a common cause of respiratory tract infections (RTI in children. We recently demonstrated that this bacterium can be carried asymptomatically in the respiratory tract of children. To identify potential genetic differences between M. pneumoniae strains that are carried asymptomatically and those that cause symptomatic infections, we performed whole-genome sequence analysis of 20 M. pneumoniae strains. The analyzed strains included 3 reference strains, 3 strains isolated from asymptomatic children, 13 strains isolated from clinically well-defined patients suffering from an upper (n=4 or lower (n=9 RTI, and one strain isolated from a follow-up patient who recently recovered from an RTI. The obtained sequences were each compared to the sequences of the reference strains. To find differences between strains isolated from asymptomatic and symptomatic individuals, a variant comparison was performed between the different groups of strains. Irrespective of the group (asymptomatic versus symptomatic from which the strains originated, subtype 1 and subtype 2 strains formed separate clusters. We could not identify a specific genotype associated with M. pneumoniae virulence. However, we found marked genetic differences between clinical isolates and the reference strains, which indicated that the latter strains may not be regarded as appropriate representatives of circulating M. pneumoniae strains.

  17. Comparison of Mycoplasma pneumoniae Genome Sequences from Strains Isolated from Symptomatic and Asymptomatic Patients

    Science.gov (United States)

    Spuesens, Emiel B. M.; Brouwer, Rutger W. W.; Mol, Kristin H. J. M.; Hoogenboezem, Theo; Kockx, Christel E. M.; Jansen, Ruud; Van IJcken, Wilfred F. J.; Van Rossum, Annemarie M. C.; Vink, Cornelis

    2016-01-01

    Mycoplasma pneumoniae is a common cause of respiratory tract infections (RTIs) in children. We recently demonstrated that this bacterium can be carried asymptomatically in the respiratory tract of children. To identify potential genetic differences between M. pneumoniae strains that are carried asymptomatically and those that cause symptomatic infections, we performed whole-genome sequence analysis of 20 M. pneumoniae strains. The analyzed strains included 3 reference strains, 3 strains isolated from asymptomatic children, 13 strains isolated from clinically well-defined patients suffering from an upper (n = 4) or lower (n = 9) RTI, and one strain isolated from a follow-up patient who recently recovered from an RTI. The obtained sequences were each compared to the sequences of the reference strains. To find differences between strains isolated from asymptomatic and symptomatic individuals, a variant comparison was performed between the different groups of strains. Irrespective of the group (asymptomatic vs. symptomatic) from which the strains originated, subtype 1 and subtype 2 strains formed separate clusters. We could not identify a specific genotype associated with M. pneumoniae virulence. However, we found marked genetic differences between clinical isolates and the reference strains, which indicated that the latter strains may not be regarded as appropriate representatives of circulating M. pneumoniae strains. PMID:27833597

  18. Isolation and Molecular Characterization of Brucella Isolates in Cattle Milk in Uganda

    Directory of Open Access Journals (Sweden)

    Denis Rwabiita Mugizi

    2015-01-01

    Full Text Available Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle. To shed new light on the epidemiology of brucellosis in Uganda the present study used phenotypic and molecular approaches to delineate the Brucella species, biovars, and genotypes shed in cattle milk. Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda. These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world. This study further highlights the usefulness of MLVA as an epidemiological tool for investigation of Brucella infections.

  19. Isolation and Molecular Characterization of Brucella Isolates in Cattle Milk in Uganda

    Science.gov (United States)

    Mugizi, Denis Rwabiita; Muradrasoli, Shaman; Erume, Joseph; Nasinyama, George William; Waiswa, Charles; Magnusson, Ulf

    2015-01-01

    Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle. To shed new light on the epidemiology of brucellosis in Uganda the present study used phenotypic and molecular approaches to delineate the Brucella species, biovars, and genotypes shed in cattle milk. Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda. These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR) loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world. This study further highlights the usefulness of MLVA as an epidemiological tool for investigation of Brucella infections. PMID:25793204

  20. Enhanced efficacy of recombinant Brucella abortus RB51 vaccines against B. melitensis infection in mice.

    Science.gov (United States)

    Vemulapalli, Ramesh; Contreras, Andrea; Sanakkayala, Neelima; Sriranganathan, Nammalwar; Boyle, Stephen M; Schurig, Gerhardt G

    2004-09-08

    Brucella abortus strain RB51 is an attenuated rough strain, currently being used as the official live vaccine for bovine brucellosis in the USA and several other countries. In strain RB51, the wboA gene, encoding a glycosyltransferase required for the O-side chain synthesis, is disrupted by an IS711 element. Recently, we have demonstrated that strain RB51WboA, RB51 complemented with a functional wboA gene, remains rough but expresses low quantities of O-side chain in the cytoplasm. Mice vaccinated with strain RB51WboA develop greatly enhanced resistance against challenge with B. abortus virulent strain 2308. We have also demonstrated that overexpression of Cu/Zn superoxide dismutase (SOD) in strain RB51 (RB51SOD) significantly increases its vaccine efficacy against strain 2308 challenge. In this study, we constructed a new recombinant strain, RB51SOD/WboA, that over expresses SOD with simultaneous expression of O-side chain in the cytoplasm. We tested the vaccine potential of strains RB51SOD, RB51WboA, RB51SOD/WboA against challenge with virulent Brucella melitensis 16M and B. abortus 2308 in mice. In comparison with strain RB51, strain RB51SOD induced better protection against strain 2308, but not strain 16M, challenge. Similar to strain RB51WboA, vaccination with strain RB51SOD/WboA resulted in complete protection of the mice from infection with strain 2308. When challenged with strain 16M, mice vaccinated with either strain RB51WboA or strain RB51SOD/WboA were significantly better protected than those vaccinated with strain RB51 or RB51SOD. These results suggest that strains RB51WboA and RB51SOD/WboA are good vaccine candidates for inducing enhanced protection against B. melitensis infection.

  1. Comparison of cytokine immune responses to Brucella abortus and Yersinia enterocolitica serotype O:9 infections in BALB/c mice.

    Science.gov (United States)

    Gu, Wenpeng; Wang, Xin; Qiu, Haiyan; Cui, Buyun; Zhao, Shiwen; Zheng, Han; Xiao, Yuchun; Liang, Junrong; Duan, Ran; Jing, Huaiqi

    2013-12-01

    Brucella abortus and Yersinia enterocolitica serotype O:9 serologically cross-react in the immune response with the host; therefore, our aim was to compare the immune responses to these two pathogens. We selected typical B. abortus and Y. enterocolitica O:9 strains to study the cytokine immune response and the histopathological changes in livers and spleens of BALB/c mice. The data showed the cytokine responses to the two strains of pathogens were different, where the average levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), gamma interferon (IFN-γ), interleukin-12 (IL-12), and tumor necrosis factor alpha (TNF-α) were higher with B. abortus infections than with Y. enterocolitica O:9 infections, especially for IFN-γ, while the IL-10 level was lower and the levels of IL-1β, IL-4, IL-5, and IL-6 were similar. The histopathological effects in the livers and spleens of the BALB/c mice with B. abortus and Y. enterocolitica O:9 infections were similar; however, the pathological changes in the liver were greater with B. abortus infections, while damage in the spleen was greater with Y. enterocolitica O:9 infections. These observations show that different cytokine responses and histopathological changes occur with B. abortus and Y. enterocolitica O:9 infections.

  2. Characterization of Methylobacterium strains isolated from the phyllosphere and description of Methylobacterium longum sp nov

    NARCIS (Netherlands)

    Knief, C.; Dengler, V.; Bodelier, P.L.E.; Vorholt, J.A.

    2012-01-01

    Methylobacterium strains are abundantly found in the phyllosphere of plants. Morphological, physiological and chemotaxonomical properties of 12 previously isolated strains were analyzed in order to obtain a more detailed overview of the characteristics of phyllosphere colonizing Methylobacterium

  3. Antimicrobial Activity of Starch-Degrading Lactobacillus Strains Isolated from Boza

    National Research Council Canada - National Science Library

    Petrova, Penka M; Petrov, Kaloyan K

    2011-01-01

    .... In the present study, three strains-Lactobacillus paracasei B41, Lactobacillus plantarum Bom 816, and Lactobacillus pentosus N3, isolated from boza and possessing significant amylolytic activity...

  4. Brucella abortus detected in cheese from the Amazon region: differentiation of a vaccine strain (B19) from the field strain in the states of Pará, Amapá and Rondônia, Brazil

    OpenAIRE

    Silva,Jacqueline; Moraes, Carina M. de; Cleyzer L. Silva; Gustavo A. Sales; Lara B. Keid; Paulo C.M. Matos; Lara,Ana P.S.S.; Moraes,Carla C.G. de

    2016-01-01

    Abstract: Brucellosis is an infectious-contagious disease responsible for significant economic losses to the meat and milk supply chain, because it causes reproductive disorders in animals and is a chronic anthropozoonosis. This study was designed to detect the DNA of Brucella spp. in cheese and to differentiate between a vaccine strain (B19) and the field strain. Sixty-six samples of different cheeses which are produced and marketed in three states of the Brazilian Amazon region (Amapá [5 sa...

  5. Molecular characterization of Mycobacterium bovis strains isolated from cattle slaughtered at two abattoirs in Algeria

    Directory of Open Access Journals (Sweden)

    Ouzrout Rachid

    2009-01-01

    Full Text Available Abstract Background Bovine Tuberculosis is prevalent in Algeria despite governmental attempts to control the disease. The objective of this study was to conduct, for the first time, molecular characterization of a population sample of Mycobacterium bovis strains isolated from slaughter cattle in Algeria. Between August and November 2007, 7250 animals were consecutively screened at the abattoirs of Algiers and Blida. In 260 animals, gross visible granulomatous lesions were detected and put into culture. Bacterial isolates were subsequently analysed by molecular methods. Results Altogether, 101 bacterial strains from 100 animals were subjected to molecular characterization. M. bovis was isolated from 88 animals. Other bacteria isolated included one strain of M. caprae, four Rhodococcus equi strains, three Non-tuberculous Mycobacteria (NTM and five strains of other bacterial species. The M. bovis strains isolated showed 22 different spoligotype patterns; four of them had not been previously reported. The majority of M. bovis strains (89% showed spoligotype patterns that were previously observed in strains from European cattle. Variable Number of Tandem Repeat (VNTR typing supported a link between M. bovis strains from Algeria and France. One spoligotype pattern has also been shown to be frequent in M. bovis strains from Mali although the VNTR pattern of the Algerian strains differed from the Malian strains. Conclusion M. bovis infections account for a high amount of granulomatous lesions detected in Algerian slaughter cattle during standard meat inspection at Algiers and Blida abattoir. Molecular typing results suggested a link between Algerian and European strains of M. bovis.

  6. Molecular characterization of Mycobacterium bovis strains isolated from cattle slaughtered at two abattoirs in Algeria

    Science.gov (United States)

    Sahraoui, Naima; Müller, Borna; Guetarni, Djamel; Boulahbal, Fadéla; Yala, Djamel; Ouzrout, Rachid; Berg, Stefan; Smith, Noel H; Zinsstag, Jakob

    2009-01-01

    Background Bovine Tuberculosis is prevalent in Algeria despite governmental attempts to control the disease. The objective of this study was to conduct, for the first time, molecular characterization of a population sample of Mycobacterium bovis strains isolated from slaughter cattle in Algeria. Between August and November 2007, 7250 animals were consecutively screened at the abattoirs of Algiers and Blida. In 260 animals, gross visible granulomatous lesions were detected and put into culture. Bacterial isolates were subsequently analysed by molecular methods. Results Altogether, 101 bacterial strains from 100 animals were subjected to molecular characterization. M. bovis was isolated from 88 animals. Other bacteria isolated included one strain of M. caprae, four Rhodococcus equi strains, three Non-tuberculous Mycobacteria (NTM) and five strains of other bacterial species. The M. bovis strains isolated showed 22 different spoligotype patterns; four of them had not been previously reported. The majority of M. bovis strains (89%) showed spoligotype patterns that were previously observed in strains from European cattle. Variable Number of Tandem Repeat (VNTR) typing supported a link between M. bovis strains from Algeria and France. One spoligotype pattern has also been shown to be frequent in M. bovis strains from Mali although the VNTR pattern of the Algerian strains differed from the Malian strains. Conclusion M. bovis infections account for a high amount of granulomatous lesions detected in Algerian slaughter cattle during standard meat inspection at Algiers and Blida abattoir. Molecular typing results suggested a link between Algerian and European strains of M. bovis. PMID:19173726

  7. Decolorization of Methyl Red by Staphylococcus saprophyticus strain AUCASVE3 Isolated from Textile Effluent

    OpenAIRE

    Azizul Hakim; Sirajul Hoque; Shah Mohammad Ullah

    2014-01-01

    In this study, an attempt was made to isolate methyl red, an azo dye, decolorizing bacteria from textile effluent. Among three isolates with methyl red decolorizing ability, the potential bacterial isolates, Staphylococcus saprophyticus strain AUCASVE3 was selected for identification and characterization. The organism was found to belong to the genus Staphylococcus and species saprophyticus. The isolate Staphylococcus saprophyticus strain AUCASVE3 could decolorize 600, 700, 800, and 1000 mg/L...

  8. Characteristics of Bacterial Strains from Pseudomonas Genera Isolated from Diseased Plum Trees

    Directory of Open Access Journals (Sweden)

    Veljko Gavrilović

    2008-01-01

    Full Text Available Characteristics of Pseudomonas syringae strains isolated from diseased plum trees are presented is this paper. Based on pathogenic, biochemical and physiological characteristics, isolated starins were divided into two groups: First group of strains, isolated from diseased plum branches with symptoms of suden decay, was simillar to Pseudomonas syringae pv. syringae; second group of strains, isolated from necrotic flower buds on plum trees, exhibited characteristics simillar to Pseudomonas syringae pv. morsprunorum. In addition, phytopathogenic fungi belonging to genera Phomopsis, Botryosphaeria and Leucostoma, were also isolated from diseased plum trees. Further study of these pathogens and their role in the epidemiology of suden plum trees decay is in progress.

  9. Selective enrichment media bias the types of Salmonella enterica strains isolated from mixed strain cultures and complex enrichment broths.

    Science.gov (United States)

    Gorski, Lisa

    2012-01-01

    For foodborne outbreak investigations it can be difficult to isolate the relevant strain from food and/or environmental sources. If the sample is contaminated by more than one strain of the pathogen the relevant strain might be missed. In this study mixed cultures of Salmonella enterica were grown in one set of standard enrichment media to see if culture bias patterns emerged. Nineteen strains representing four serogroups and ten serotypes were compared in four-strain mixtures in Salmonella-only and in cattle fecal culture enrichment backgrounds using Salmonella enrichment media. One or more strain(s) emerged as dominant in each mixture. No serotype was most fit, but strains of serogroups C2 and E were more likely to dominate enrichment culture mixtures than strains of serogroups B or C1. Different versions of Rappaport-Vassiliadis (RV) medium gave different patterns of strain dominance in both Salmonella-only and fecal enrichment culture backgrounds. The fittest strains belonged to serogroups C1, C2, and E, and included strains of S. Infantis, S. Thompson S. Newport, S. 6,8:d:-, and S. Give. Strains of serogroup B, which included serotypes often seen in outbreaks such as S. Typhimurium, S. Saintpaul, and S. Schwarzengrund were less likely to emerge as dominant strains in the mixtures when using standard RV as part of the enrichment. Using a more nutrient-rich version of RV as part of the protocol led to a different pattern of strains emerging, however some were still present in very low numbers in the resulting population. These results indicate that outbreak investigations of food and/or other environmental samples should include multiple enrichment protocols to ensure isolation of target strains of Salmonella.

  10. Oxidative metabolic profiles of Brucella strains isolated from marine mammals: contribution to their species classification.

    Science.gov (United States)

    Jacques, Isabelle; Grayon, Maggy; Verger, Jean-Michel

    2007-05-01

    Since the 1990s, Brucella strains not matching the characteristics of any of the six conventional species have been isolated worldwide from marine mammals. In this study, 31 Brucella strains isolated from various marine mammals were examined for their oxidative metabolic pattern on 12 amino-acid and carbohydrate substrates. Three main oxidative profiles different from those of the Brucella terrestrial mammal strains were identified for the marine mammal strains: one gathering strains isolated from pinnipeds and two gathering strains from cetaceans. Thus, both oxidative metabolism results and previous molecular studies are in agreement with the proposal of two new Brucella species, Brucella pinnipediae and Brucella cetaceae, to classify the Brucella strains isolated from marine mammals, and are also in accordance with a classification of species of the Brucella genus based on host preference.

  11. Complement Fixation Test To Assess Humoral Immunity in Cattle and Sheep Vaccinated with Brucella abortus RB51

    Science.gov (United States)

    Adone, Rosanna; Ciuchini, Franco

    1999-01-01

    The live attenuated Brucella abortus strain RB51 is a rifampin-resistant, lipopolysaccharide (LPS) O-chain-deficient mutant of virulent B. abortus 2308. The reduced O-chain content in RB51 prevents this bacterium from inducing antibodies detectable by the conventional serologic tests for bovine brucellosis diagnosis that mainly identify antibodies to LPS. The absence of available serologic tests for RB51 also complicates the diagnosis of possible RB51 infections in humans exposed to this strain. The purpose of this study was to evaluate the suitability of a complement fixation (CF) test performed with the rough strain B. abortus RB51, previously deprived of anticomplementary activity, in detecting anti-B. abortus RB51 antibodies in cattle and sheep experimentally vaccinated with this strain. The results of this study showed that a CF test with RB51 as the antigen is able to specifically detect antibodies following RB51 vaccination in cattle and sheep. In addition, this method could be a useful tool for detecting B. abortus RB51 infection in humans. PMID:10548564

  12. Heavy metal resistance by two bacteria strains isolated from a ...

    African Journals Online (AJOL)

    The strain DX-T3-03 had a larger range of antibiotic resistance than DX-T3-01. On the basis of 16S rDNA sequencing, the two strains were identified as Ralstonia pickettii strain DX-T3-01 and Sphingomonas sp. strain DX-T3-03, respectively. This study supplied potential bacterial materials for tailing bioremediation in the ...

  13. ANTIBIOTIC RESISTANCE IN ENTEROBACTERIACEAE STRAINS ISOLATED FROM CHICKEN AND MILK SAMPLES

    Directory of Open Access Journals (Sweden)

    Lukáš Hleba

    2015-02-01

    Full Text Available Antibiotic resistance and identification of strains in Enterobacteriaceae genera isolated from milk, milk products and rectal swabs of chicken was examined in this experiment. After samples collection cultivation and identification of bacterial strain was done. MALDI TOF MS Biotyper for identification of Enterobacteriaceae strains was used. For susceptibility testing disc diffusion methodology was used according by EUCAST. Results showed high level of ampicillin resistance in isolates from milk and milk samples. The highest streptomycin resistance was detected in isolates from rectal swabs of chicken. After identification, we determined that S. enterica ser. Typhimurium, which was isolated from rectal swabs of chicken showed the most multi-resistance from all identificated strains of Enterobacteriaceae. The most isolates bacterial strain was E. coli, which showed resistance against four antibiotics from rectal swabs of chicken. Also our results showed that the higher resistance level is in rectal swabs of chicken like in milk samples.

  14. Physiology of Saccharomyces cerevisiae strains isolated from Brazilian biomes: new insights into biodiversity and industrial applications

    DEFF Research Database (Denmark)

    Beato, Felipe B.; Bergdahl, Basti; Rosa, Carlos A.

    2016-01-01

    Fourteen indigenous Saccharomyces cerevisiae strains isolated from the barks of three tree species located in the Atlantic Rain Forest and Cerrado biomes in Brazil were genetically and physiologically compared to laboratory strains and to strains from the Brazilian fuel ethanol industry. Although...

  15. Genome Sequences of Two Pseudoalteromonas Strains Isolated from the South China Sea

    Science.gov (United States)

    Zeng, Zhenshun; Dai, Shikun; Xie, Yunchang; Tian, Xinpeng; Li, Jie

    2014-01-01

    Two Pseudoalteromonas strains, SCSIO 04301 and SCSIO 11900, were isolated from the South China Sea, and both strains form biofilms. Here we present the draft genome sequences of these two strains, which will aid the study of marine microbes that are adapted to marine sediments or are associated with eukaryotic hosts. PMID:24744335

  16. Coagulase-negative staphylococci strains resistant to oxacillin isolated from neonatal blood cultures

    Directory of Open Access Journals (Sweden)

    Valeria Cataneli Pereira

    2013-11-01

    Full Text Available Coagulase-negative staphylococci (CoNS are the microorganisms most frequently isolated from clinical samples and are commonly found in neonatal blood cultures. Oxacillin is an alternative treatment of choice for CoNS infections; however, resistance to oxacillin can have a substantial impact on healthcare by adversely affecting morbidity and mortality. The objective of this study was to detect and characterise oxacillin-resistant CoNS strains in blood cultures of newborns hospitalised at the neonatal ward of the University Hospital of the Faculty of Medicine of Botucatu. One hundred CoNS strains were isolated and the mecA gene was detected in 69 of the CoNS strains, including 73.2% of Staphylococcus epidermidis strains, 85.7% of Staphylococcus haemolyticus strains, 28.6% of Staphylococcus hominis strains and 50% of Staphylococcus lugdunensis strains. Among these oxacillin-resistant CoNS strains, staphylococcal cassette chromosome mec (SCCmec type I was identified in 24.6%, type II in 4.3%, type III in 56.5% and type IV in 14.5% of the strains. The data revealed an increase in the percentage of CoNS strains isolated from blood cultures from 1991-2009. Furthermore, a predominant SCCmec profile of the oxacillin-resistant CoNS strains isolated from neonatal intensive care units was identified with a prevalence of SCCmec types found in hospital-acquired strains.

  17. Genetic analysis of Saccharomyces cerevisiae strains isolated from palm wine in eastern Nigeria. Comparison with other African strains.

    Science.gov (United States)

    Ezeronye, O U; Legras, J-L

    2009-05-01

    To study the yeast diversity of Nigerian palm wines by comparison with other African strains. Twenty-three Saccharomyces cerevisiae strains were obtained from palm wine samples collected at four locations in eastern Nigeria, and characterized using different molecular techniques: internal transcribed spacer restriction fragment length polymorphism and sequence analysis, pulsed field gel electrophoresis, inter delta typing and microsatellite multilocus analysis. These techniques revealed that palm wine yeasts represent a group of closely related strains that includes other West African isolates (CBS400, NCYC110, DVPG6044). Population analysis revealed an excess of homozygote strains and an allelic richness similar to wine suggestive of local domestication. Several other African yeast strains were not connected to this group. Ghana sorghum beer strains and other African strains (DBVPG1853 and MUCL28071) displayed strikingly high relatedness with European bread, beer or wine strains, and the genome of strain MUCL30909 contained African and wine-type alleles, indicating its hybrid origin. Nigerian palm wine yeast represents a local specific yeast flora, whereas a European origin or hybrid was suspected for several other Africa isolates. This study presents the first genetic characterization of an autochthonous African palm wine yeast population and confirms the idea that human intervention has favoured yeast migration.

  18. Genomic DNA restriction endonuclease from Pasteurella multocida isolated from Indonesia, katha strain and reference strains and analysed by PFGE

    Directory of Open Access Journals (Sweden)

    Supar

    2003-10-01

    Full Text Available Pasteurella multocida strains are the causative disease agents of wide range of domestic and wild animals in Indonesia. The most important serotypes are associated with Hemorrhagic septicaemic (HS diseases in cattle and buffaloes, cholera in ducks and chickens. The HS disease associated with P. multocia in large ruminants in Indonesia is controled by killed whole cell vaccines produced by the use of P. multocida Katha strains. There is no discriminatory data of the molecular biology technique has been applied to investigate P. multocida isolates from different geographic locations in Indonesia. The purpose of this studies were to observe the genetic diversity among P. multocida isolated from various geograpic locations and compared with Katha vaccine strain and other reference strains. A total samples of 38 isolates and strains of P. multocida were analysed by means of pulsed-field gel electrophoresis (PFGE. Each sample was grown in nutrient broth, cells were separeted by centrifugation. Whole cell pellet was mixed with agarose and then prepared agarose plugs. The genomic DNA of each sample was digested in situ (plug with either restriction endonuclease of ApaI and/or BamHI. The digested genomic DNA of each sample was analysed by PFGE, the genomic DNA restricted profile of each sample was compared with others. The use of ApaI restriction endonuclease digestion and analysed by PFGE, demonstrated that 34 out of 38 P. multocia samples could be differentiated into 16 ApaI types, whereas based on the BamHI digestion of these samples were differentiated into 20 BamHI types. Genomic DNA restriction pattern of Indonesian P. multocida isolates originated from cattle and buffaloes associated with haemorrhagic septicaemic diseases demonstrated different pattern to those of vaccine Katha strain, poultry strains as well as the reference strains currenly kept at Balitvet Culture Collection (BCC unit. Two P. multocida isolates derived from ducks with cholera

  19. Monoclonal antibodies to Toxoplasma gondii strain 119 identify recently isolated Danish strains as one group

    DEFF Research Database (Denmark)

    Jensen, L.; Petersen, E.; Henriksen, S.A.

    1998-01-01

    Four mAb raised against the Danish Toxoplasma gondii strain 119, were selected by screening hybridoma supernatants by indirect immunofluorescence against tachyzoites of the RH strain in order to obtain strain restricted markers. Strain restriction extended beyond discrimination of the 119 and RH...

  20. Diversity of Vibrio navarrensis Revealed by Genomic Comparison: Veterinary Isolates Are Related to Strains Associated with Human Illness and Sewage Isolates While Seawater Strains Are More Distant.

    Science.gov (United States)

    Schwartz, Keike; Kukuc, Cindy; Bier, Nadja; Taureck, Karin; Hammerl, Jens A; Strauch, Eckhard

    2017-01-01

    Strains of Vibrio navarrensis are present in aquatic environments like seawater, rivers, and sewage. Recently, strains of this species were identified in human clinical specimens. In this study, V. navarrensis strains isolated from livestock in Germany were characterized that were found in aborted fetuses and/or placentas after miscarriages. The veterinary strains were analyzed using phenotypical and genotypical methods and compared to isolates from marine environments of the Baltic Sea and North Sea. The investigated phenotypical traits were similar in all German strains. Whole genome sequencing (WGS) was used to evaluate a phylogenetic relationship by performing a single nucleotide polymorphism (SNP) analysis. For the SNP analysis, WGS data of two American human pathogenic strains and two Spanish environmental isolates from sewage were included. A phylogenetic analysis of concatenated sequences of five protein-coding housekeeping genes (gyrB, pyrH, recA, atpA, and rpoB), was additionally performed. Both phylogenetic analyses reveal a greater distance of the environmental seawater strains to the other strains. The phylogenetic tree constructed from concatenated sequences of housekeeping genes places veterinary, human pathogenic and Spanish sewage strains into one cluster. Presence and absence of virulence-associated genes were investigated based on WGS data and confirmed by PCR. However, this analysis showed no clear pattern for the potentially pathogenic strains. The detection of V. navarrensis in human clinical specimens strongly suggests that this species should be regarded as a potential human pathogen. The identification of V. navarrensis strains in domestic animals implicates a zoonotic potential of this species. This could indicate a potential threat for humans, as according to the "One Health" concept, human, animal, and environmental health are linked. Future studies are necessary to search for reservoirs of these bacteria in the environment and/or in

  1. Development of a Selective Culture Medium for Primary Isolation of the Main Brucella Species▿

    OpenAIRE

    Miguel, María Jesús de; Marín, Clara M.; Muñoz, Pilar M.; Dieste, L.; Grilló, María Jesús; Blasco, José M

    2011-01-01

    Bacteriological diagnosis of brucellosis is performed by culturing animal samples directly on both Farrell medium (FM) and modified Thayer-Martin medium (mTM). However, despite inhibiting most contaminating microorganisms, FM also inhibits the growth of Brucella ovis and some B. melitensis and B. abortus strains. In contrast, mTM is adequate for growth of all Brucella species but only partially inhibitory for contaminants. Moreover, the performance of both culture media for isolating B. suis ...

  2. Probiotic properties of Lactobacillus strains isolated from Tibetan kefir grains

    National Research Council Canada - National Science Library

    Zheng, Yongchen; Lu, Yingli; Wang, Jinfeng; Yang, Longfei; Pan, Chenyu; Huang, Ying

    2013-01-01

    ...) isolated from Tibetan kefir grains. Three Lactobacillus isolates identified as Lactobacillus acidophilus LA15, Lactobacillus plantarum B23 and Lactobacillus kefiri D17 that showed resistance to acid and bile salts were selected...

  3. Isolation and Characterization of Bacillus thuringiensis strains from ...

    African Journals Online (AJOL)

    Twelve isolates of Bacillus thuringiensis were obtained from soil samples collected from farmers' fields in Kakamega and Machakos districts. Soil samples from Machakos yielded more B. thuringiensis isolates t han those from Kakamega. Three isolates K13-1, 12F-K and K10-2 were obtained from Kakamega soil samples.

  4. Full genome sequence of a Danish isolate of Mycobacterium avium subspecies paratuberculosis, strain Ejlskov2007

    DEFF Research Database (Denmark)

    Afzal, Mamuna; Abidi, Soad; Mikkelsen, Heidi

    We have sequenced a Danish isolate of Mycobacterium avium subspecies paratuberculosis, strain Ejlskov2007. The strain was isolated from faecal material of a 48 month old second parity Danish Holstein cow, with clinical symptoms of chronic diarrhoea and emaciation. The cultures were grown on Löwen......We have sequenced a Danish isolate of Mycobacterium avium subspecies paratuberculosis, strain Ejlskov2007. The strain was isolated from faecal material of a 48 month old second parity Danish Holstein cow, with clinical symptoms of chronic diarrhoea and emaciation. The cultures were grown......, consisting of 4317 unique gene families. Comparison with M. avium paratuberculosis strain K10 revealed only 3436 genes in common (~70%). We have used GenomeAtlases to show conserved (and unique) regions along the Ejlskov2007 chromosome, compared to 2 other Mycobacterium avium sequenced genomes. Pan......-genome analyses of the sequenced Mycobacterium genomes reveal a surprisingly open and diverse set of genes for this bacterial genera....

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

    Directory of Open Access Journals (Sweden)

    Elena Shevtsova

    Full Text Available 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

  6. 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.

  7. Genotypic Characterization of Seven Strains of Mycoplasma fermentans Isolated from Synovial Fluids of Patients with Arthritis

    Science.gov (United States)

    Schaeverbeke, Thierry; Clerc, Maïthé; Lequen, Laurence; Charron, Alain; Bébéar, Cécile; de Barbeyrac, Bertille; Bannwarth, Bernard; Dehais, Joël; Bébéar, Christiane

    1998-01-01

    We performed a genotypic characterization of seven strains of Mycoplasma fermentans which have been isolated from the synovial fluid of patients with rheumatoid arthritis (n = 2), spondyloarthropathy (n = 1), and unclassified arthritis (n = 4). We compared them to three reference strains (strains PG18 and K7 and incognitus strain) and to a clinical isolate from the urethra of a patient with nongonococcal urethritis. The characterization methods included electrophoresis of native DNA, arbitrarily primed PCR, and restriction fragment length polymorphism analysis following conventional and pulsed-field gel electrophoresis. Southern blot analysis with a probe internal to an insertion sequence was performed with the restriction products produced by the last two techniques. No extrachromosomal DNA sequences were detected. The M. fermentans strains identified by these methods did not present a unique profile, but they could be separated into two main categories: four articular isolates were genetically related to PG18 and the three other isolates, the urethral isolate, and the incognitus strain were related to K7. We also looked for the presence of the bacteriophage MAV1 (associated with the arthritogenic property of Mycoplasma arthritidis in rodents) in the M. fermentans strains. MAV1 DNA was not detected in either the clinical isolates or the reference strains of M. fermentans. PMID:9574681

  8. Characteristics of Yersinia enterocolitica biotype 1A strains isolated from patients and asymptomatic carriers.

    Science.gov (United States)

    Stephan, R; Joutsen, S; Hofer, E; Säde, E; Björkroth, J; Ziegler, D; Fredriksson-Ahomaa, M

    2013-07-01

    Yersinia enterocolitica biotype 1A strains are frequently isolated from the environment, foods, and animals, and also from humans with yersiniosis. There are controversial reports on the pathogenicity of biotype 1A strains. In this study, 811 fecal samples from asymptomatic humans from Switzerland were studied for the presence of Y. enterocolitica. Nine (1.1%) of the 811 samples were positive for Y. enterocolitica 1A. These strains were compared with 12 Y. enterocolitica 1A strains from Swiss patients with diarrhea isolated in the same year. Almost all (20/21) Y. enterocolitica 1A strains carried the ystB gene, seven strains carried the hreP gene, and none carried the ail, ystA, myfA, yadA, or virF genes. Most (17/21) Y. enterocolitica 1A strains belonged to two major clusters, A and B, by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Strains of cluster B were only isolated from humans with diarrhea; however, ystB and hreP genes were detected in strains from both clinical and non-clinical samples and from strains of clusters A and B. Using ribotyping, six restriction patterns among biotype 1A strains were obtained with HindIII enzyme. The most common ribotype (RT I) was found in strains isolated from humans with and without diarrhea. All biotype 1A strains had a unique NotI profile by pulsed-field gel electrophoresis (PFGE), showing a very high genetic diversity. In this study, Y. enterocolitica 1A strains from clinical and non-clinical samples could not be clearly differentiated from each other. More research is needed in order to prove that biotype 1A strains are a primary cause for human yersiniosis and not only a secondary finding.

  9. Streptococcus anginosus (milleri) Group Strains Isolated in Poland (1996-2012) and their Antibiotic Resistance Patterns.

    Science.gov (United States)

    Obszańska, Katarzyna; Kern-Zdanowicz, Izabella; Kozińska, Aleksandra; Machura, Katarzyna; Stefaniuk, Elzbieta; Hryniewicz, Waleria; Sitkiewicz, Izabela

    2016-01-01

    Streptococcus anginosus, Streptococcus intermedius and Streptococcus constellatus form a group of related streptococcal species, namely the Streptococcus Anginosus Group (SAG). The group, previously called "milleri" had been rarely described until 1980/1990 as source of infections. Nowadays SAG bacteria are often described as pathogens causing predominantly purulent infections. The number of infections is highly underestimated, as SAG strains are often classified in the microbiology laboratory as less virulent "viridans streptococci" Epidemiological situation regarding SAG infections in Poland has been unrecognized, therefore we performed a retrospective analysis of strains isolated between 1996 and 2012. Strains suspected of belonging to SAG were re-identified using an automated biochemical approach (Vitek2) and MALDI-TOF MS. We performed first analysis of antibiotic resistance among SAG strains isolated in Poland using automated methods (Vitek2), disk diffusion tests and E-Tests. We also performed PCR detection of resistance determinants in antibiotic resistant strains. Clonal structure of analyzed strains was evaluated with PFGE and MLVF methods. All three species are difficult to distinguish using automated diagnostic methods and the same is true for automated MIC evaluation. Our analysis revealed SAG strains are rarely isolated in Poland, predominantly from purulent infections. All isolates are very diverse on the genomic level as estimated by PFGE and MLVF analyses. All analyzed strains are sensitive to penicillin, a substantial group of strains is resistant to macrolides and the majority of strains are resistant to tetracycline.

  10. Formation of biofilm by strains of Listeria monocytogenes isolated ...

    African Journals Online (AJOL)

    Quantification of biofilm formation by 40 Listeria monocytogenes strains from wara soft cheese and its processing environment was assessed on glass vials surfaces. Attachement to glass surface was quantified using a crystal violet binding assay. All the 40 strains produced biofilms after 48 and 72 h incubation at 37oC.

  11. MICROBIOLOGICAL CHARACTERISATION OF Haemophilus influenzae STRAINS ISOLATED FROM PATIENTS WITH INVASIVE AND RESPIRATORY DISEASES

    Directory of Open Access Journals (Sweden)

    Tomislav Kostyanev

    2010-01-01

    Full Text Available A total of 175 H. influenzae strains were collected between 1994 and 2009 from all aged patient groups. The strains were isolated from patients with invasive and community-acquired respiratory tract infections. All strains were identified according to standard microbiological methods. Serotyping was done by a coagglutination test and by molecular PCR capsular genotyping. Beta-lactamase production was determined by the chromogenic cephalosporin test with nitrocephin as substrate. Most of the isolated H. influenzae strains were from children under 5 years of age (57.7%. Overall, 61 strains belonged to serotype b (34.9% by the means of PCR capsular typing, 1 strain was type f, and 113 isolates (64.6% were non-typeable (non-encapsulated H. influenzae. Among the infants and children with meningitis or other invasive infections, aged 2 month to 5 years, all strains, except one, were serotype b. In respiratory tract infections (pneumonia, otitis media, sinusitis and people with chronic pulmonary diseases - exacerbations of COPD, bronchiectasis, cystic fibrosis the most common - 96.5% were non-typeable strains in both groups children and adults. Overall, the prevalence of beta-lactamase production was 19.4%. But, it was much higher for invasive strains from CSF isolates - 37.7%, 25% in blood samples, and 37.5% in otitis media causative strains. Beta-lactamase production was less frequent in respiratory tract isolates - in sputum 13.3% and in URT samples - 2.3%. The rate of beta-lactamase production in CSF isolates has not changed for the last 10 years.PCR capsular genotyping method has to be performed for all non-b-type strains. The implementation of Hib vaccine in our country will be accompanied by a reduction in invasive diseases caused by H. influenzae type b in children, but it is not useful in preventing infections caused by non-typeable H. influenzae strains.

  12. Validation of a PCR Assay for Chlamydophila abortus rRNA gene detection in a murine model

    Directory of Open Access Journals (Sweden)

    Francielle Gibson da Silva-Zacarias

    2009-11-01

    Full Text Available Chlamydophila abortus (C. abortus is associated with reproductive problems in cattle, sheep, and goats. Diagnosis of C. abortus using embryonated chicken eggs or immortalized cell lines has a very low sensitivity. Polymerase chain reaction (PCR assays have been used to detect C. abortus infection in clinical specimens and organ fragments, such as placenta, fetal organs, vaginal secretions, and semen. The aim of this study was to develop a PCR assay for the amplification of an 856-bp fragment of the rRNA gene of the Chlamydiaceae family. The PCR assay was evaluated using organs from 15 mice experimentally infected with the S26/3 reference strain of C. abortus. The results of the rRNA PCR were compared to the results from another PCR system (Omp2 PCR that has been previously described for the Omp2 (outer major protein gene from the Chlamydiaceae family. From the 15 C. abortus-inoculated mice, 13 (K=0.84, standard error =0.20 tested positive using the rRNA PCR assay and 9 (K=0.55, standard error=0.18 tested positive using the Omp2 PCR assay. The detection limit, measured using inclusion-forming units (IFU, for C. abortus with the rRNA PCR (1.05 IFU was 100-fold lower than for the Omp2 PCR (105 IFU. The higher sensitivity of the rRNA PCR, as compared to the previously described PCR assay, and the specificity of the assay, demonstrated using different pathogenic microorganisms of the bovine reproductive system, suggest that the new PCR assay developed in this study can be used for the molecular diagnosis of C. abortus in abortion and other reproductive failures in bovines, caprines, and ovines.Chlamydophila abortus (C. abortus é frequentemente associada a distúrbios reprodutivos em bovinos, ovinos e caprinos. Para o diagnóstico, os métodos de cultivo em ovo embrionado de galinha e em células de linhagem contínua apresentam baixa sensibilidade. A reação em cadeia da polimerase (PCR tem sido utilizada em placenta, órgãos fetais, secre

  13. Expression of curli by Escherichia coli O157:H7 strains isolated from patients during outbreaks is different from similar strains isolated from leafy green production environments

    Directory of Open Access Journals (Sweden)

    Subbarao Venkata Ravva

    2016-12-01

    Full Text Available We previously reported that the strains of Escherichia coli O157:H7 (EcO157 that survived longer in austere soil environment lacked expression of curli, a fitness trait linked with intestinal colonization. In addition, the proportion of curli-positive variants of EcO157 decreased with repeated soil exposure. Here we evaluated 84 and 176 clinical strains from outbreaks and sporadic infections in the US, plus 211 animal fecal and environmental strains for curli expression. These shiga-toxigenic strains were from 328 different genotypes, as characterized by multi-locus variable-number tandem-repeat analysis (MLVA. More than half of the fecal strains (human and animal and a significant proportion of environmental isolates (82% were found to lack curli expression. EcO157 strains from several outbreaks linked with the consumption of contaminated apple juice, produce, hamburgers, steak and beef were also found to lack curli expression. Phylogenetic analysis of fecal strains indicates curli expression is distributed throughout the population. However, a significant proportion of animal fecal isolates (84% gave no curli expression compared to human fecal isolates (58%. In addition, analysis of environmental isolates indicated nearly exclusive clustering of curli expression to a single branch of the minimal spanning tree. This indicates that curli expression depends primarily upon the type of environmental exposure and the isolation source, although genotypic differences also contribute to clonal variation in curli. Furthermore, curli-deficient phenotype appears to be a selective trait for survival of EcO157 in agricultural environments.

  14. Urea hydrolysis can predict the potential pathogenicity of Vibrio parahaemolyticus strains isolated in the Pacific Northwest.

    Science.gov (United States)

    Kaysner, C A; Abeyta, C; Trost, P A; Wetherington, J H; Jinneman, K C; Hill, W E; Wekell, M M

    1994-01-01

    The ability of some strains of Vibrio parahaemolyticus to hydrolyze urea (uh+) can be used as a marker to predict which strains isolated from molluscan shellfish harvested in the Pacific Northwest are potentially pathogenic. The thermostable direct hemolysin-producing (TDH+) characteristic is a marker that is correlated with potential pathogenicity, and all of the TDH+ strains that we have isolated have been found to be uh+. Most of the uh+ strains belong to somatic antigen groups O3, O4 and O5. TDH+ strains are usually members of groups O4 and O5. The strains most often associated with human illness are members of the uh+, O4 group. The test for urease production is a simple screening test that can be helpful in predicting which strains are potentially pathogenic. PMID:8085837

  15. EVALUATION OF THERMOTOLERANT ACETOBACTER PASTEURIANUS STRAINS ISOLATED FROM MOROCCAN FRUITS CATALYZING OXIDATIVE FERMENTATION AT HIGH TEMPERATURE.

    Science.gov (United States)

    Mounir, M; Shafiei, R; Zarmehrkhorshid, R; Hamouda, A; Alaoui, M Ismaili; Thonart, P

    2015-01-01

    Six strains of acetic acid bacteria were isolated from Moroccan local products and their potential as industrial strains was evaluated in lab-bioreactor. Three of them, namely TAV01, AF01 and CV01, isolated from traditional apple vinegar, apple and cactus fruit, respectively were selected and their responses to high temperature were assessed. Morphological and biochemical identification confirmed that these strains belong to Acetobacter species. Their growth and acetic acid production were compared with the thermoresistant reference strain, Acetobacter senegalensis and mesophilic strains of Acetobacter pasteurianus. The two strains AF01 and CV01 showed abundant growth and noticeable acetic acid production ability at high temperatures (38 to 41°C). A thermophilic character was observed for AF01 strain. Indeed, this bacterium grew better at 38 than 30°C.

  16. Cometabolic transformation and cleavage of nitrodiphenylamines by three newly isolated sulfate-reducing bacterial strains.

    Science.gov (United States)

    Drzyzga, O; Schmidt, A; Blotevogel, K

    1996-05-01

    Three sulfate-reducing bacterial strains (Desulfovibrio sp. strain SHV, Desulfococcus sp. strain WHC, and Desulfomicrobium sp. strain WHB) with the capacity to cometabolize 2-nitrodiphenylamine, 4-nitrodiphenylamine, and 2,4-dinitrodiphenylamine were newly isolated. Before breaking down the diphenylamine structure, these strains cometabolically reduce the nitrodiphenylamines to the corresponding aminodiphenylamines during anaerobic oxidation of the growth substrate lactate (Desulfovibrio strain SHV and Desulfomicrobium strain WHC) or benzoate (Desulfococcus strain WHB), leading to the formation of aniline and a smaller quantity of methylaniline. These compounds were not further metabolized by the sulfate reducers. The anaerobic metabolism of aminodiphenylamines also led to the formation of heterocyclic condensation products such as phenazine and acridine derivatives, provided that they contained an amino group in the ortho position of the diphenylamine (e.g., 2-aminodiphenylamine or 2,4-diaminodiphenylamine). In addition, low levels of indole and benzothiazole derivatives were identified, but these also were not further metabolized by the three sulfate-reducing strains.

  17. Champacyclin, a New Cyclic Octapeptide from Streptomyces Strain C42 Isolated from the Baltic Sea

    OpenAIRE

    Alexander Pesic; Heike I. Baumann; Katrin Kleinschmidt; Paul Ensle; Jutta Wiese; Süssmuth, Roderich D.; Imhoff, Johannes F.

    2013-01-01

    New isolates of Streptomyces champavatii were isolated from marine sediments of the Gotland Deep (Baltic Sea), from the Urania Basin (Eastern Mediterranean), and from the Kiel Bight (Baltic Sea). The isolates produced several oligopeptidic secondary metabolites, including the new octapeptide champacyclin (1a) present in all three strains. Herein, we report on the isolation, structure elucidation and determination of the absolute stereochemistry of this isoleucine/leucine (Ile/Leu = Xle) rich ...

  18. High-catalase strains of Mycobacterium kansasii isolated from water in Texas.

    OpenAIRE

    Steadham, J E

    1980-01-01

    Isolation techniques with membrane-filtered potable water samples resulted in the isolation of potentially pathogenic high-catalase strains of Mycobacterium kansasii from 8 of 19 representative outlets in a small central Texas town. Mycobacterium gordonae was isolated from all samples, and Mycobacterium fortuitum was isolated from two samples. Data on chlorine levels are presented along with a possible explanation for the unusually high numbers of mycobacteria in these potable water samples. ...

  19. ANTIBIOTIC RESISTANCE OF LACTOBACILLI STRAINS ISOLATED FROM MILK AND MILK PRODUCTS FROM MIDDLE SLOVAKIA

    Directory of Open Access Journals (Sweden)

    Lukáš Hleba

    2012-08-01

    Full Text Available The aim of this study was to determine the antibiotic susceptibility of lactobacilli strains isolated from milk and milk products from middle Slovakia. The 42 samples of milk and milk products were collected from sheep farming from middle Slovakia. MRS agar was used for this cultivation. It was done at 37 °C during 36-48 hours in CO2 box with 10 % CO2. Identification of lactobacilli strains was done by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with Biotyper software – MALDI TOF MS Biotyper. Four anitibiotics M.I.C. We used four anitibiotics M.I.C. evaluators strips for antibiotic susceptibility testing. These antibiotics were used in this experiment: erythromycin (E 256 – 0.015 µg/ml, ampicillin (AMP 256 – 0.015 µg/ml, gentamycin (CN 256 – 0.015 µg/ml and tetracycline (TE 256 – 0.015 µg/ml. We isolated 42 lactobacilli strains and identified them as Lactobacillus plantarum and Lactobacillus spp. We isolated 7 strains of Lactobacillus plantarum and 35 isolates were Lactobacillus spp. From the seven isolates of Lactobacillus plantarum 3 isolates were resistant to ampicillin (MIC 16 µg/ml in 2 isolates and MIC 12 µg/ml in one isolate and one resistant to tetracycline (MIC 16 µg/ml. From 35 isolates of Lactobacillus spp. 9 isolates were resistant to erytromycin (32 µg/ml, 10 isolates resistant to ampicillin (MIC 16 µg/ml in 4 isolates, MIC 12 µg/ml in 5 isolates and MIC 4 µg/ml in one isolate, 5 isolates resistant to tetracycline (16 µg/ml. Resistance to gentamycin was not detected. Lactobacilli isolated from milk and milk products were resistant to erytromycin (resistanece was 21.42 %, ampicillin (resistance was 30.95 % and tetracycline (resistance was 14.28 % and were sensitive in 100 % to gentamycin.

  20. Shiga Toxin-Producing Serogroup O91 Escherichia coli Strains Isolated from Food and Environmental Samples.

    Science.gov (United States)

    Feng, Peter C H; Delannoy, Sabine; Lacher, David W; Bosilevac, Joseph M; Fach, Patrick; Beutin, Lothar

    2017-09-15

    Shiga toxin-producing Escherichia coli (STEC) strains of the O91:H21 serotype have caused severe infections, including hemolytic-uremic syndrome. Strains of the O91 serogroup have been isolated from food, animals, and the environment worldwide but are not well characterized. We used a microarray and other molecular assays to examine 49 serogroup O91 strains (environmental, food, and clinical strains) for their virulence potential and phylogenetic relationships. Most of the isolates were identified to be strains of the O91:H21 and O91:H14 serotypes, with a few O91:H10 strains and one O91:H9 strain being identified. None of the strains had the eae gene, which codes for the intimin adherence protein, and many did not have some of the genetic markers that are common in other STEC strains. The genetic profiles of the strains within each serotype were similar but differed greatly between strains of different serotypes. The genetic profiles of the O91:H21 strains that we tested were identical or nearly identical to those of the clinical O91:H21 strains that have caused severe diseases. Multilocus sequence typing and clustered regularly interspaced short palindromic repeat analyses showed that the O91:H21 strains clustered within the STEC 1 clonal group but the other O91 serotype strains were phylogenetically diverse. IMPORTANCE This study showed that food and environmental O91:H21 strains have similar genotypic profiles and Shiga toxin subtypes and are phylogenetically related to the O91:H21 strains that have caused hemolytic-uremic syndrome, suggesting that these strains may also have the potential to cause severe illness. Copyright © 2017 American Society for Microbiology.

  1. Isolation and in vitro selection of actinomycetes strains as potential probiotics for aquaculture

    Directory of Open Access Journals (Sweden)

    Milagro García Bernal

    2015-02-01

    Full Text Available Aim: This study was designed to describe a series of in vitro tests that may aid the discovery of probiotic strains from actinomycetes. Materials and Methods: Actinomycetes were isolated from marine sediments using four different isolation media, followed by antimicrobial activity and toxicity assessment by the agar diffusion method and the hemolysis of human blood cells, respectively. Extracellular enzymatic production was monitored by the hydrolysis of proteins, lipids and carbohydrates. Tolerance to different pH values and salt concentrations was also determined, followed by hydrophobicity analysis and genetic identification of the most promising strains. Results: Five out of 31 isolated strains showed antimicrobial activity against three Vibrio species. Three non-hemolytic strains (N7, RL8 and V4 among these active isolates yielded positive results in hydrophobicity tests and exhibited good growth at salt concentrations ranging from 0% to 10%, except strain RL8, which required a salt concentration >0.6%. Although these strains did not grow at pH<3, they showed different enzymatic activities. Phylogenetic analysis revealed that strains N7 and V4 have more than 99% identity with several Streptomyces species, whereas the closest matches to strain RL8 are Streptomyces panacagri and Streptomyces flocculus, with 98% and 98.2% similarity, respectively. Conclusion: Three actinomycetes strains showing probiotic-like properties were discovered using several in vitro tests that can be easily implemented in different institutions around the world.

  2. Characterization and phylogenetic analysis of Varicella-zoster virus strains isolated from Korean patients.

    Science.gov (United States)

    Kim, Min Ho; Jeon, Jeong Seon; Kim, In Kyo; Park, Ji Seon; Park, Hosun; Shin, Ok Sarah; Lee, Chan Hee

    2017-08-01

    Varicella-zoster virus (VZV) is a causative agent of chickenpox in primary infection and shingles after its reactivation from latency. Complete or almost-complete genomic DNA sequences for various VZV strains have been reported. Recently, clinical VZV strains were isolated from Korean patients whose genome was sequenced using high-throughput sequencing technology. In this study, we analyzed single nucleotide polymorphism (SNP) of VZV strains to genetically characterize Korean clinical isolates. Phylogenetic analyses revealed that three Korean strains, YC01, YC02, and YC03, were linked to clade 2. Comprehensive SNP analysis identified 86 sites specific for the 5 VZV clades. VZV strains isolated from Korea did not form a phylogenetic cluster. Rather, YC02 and YC03 clustered strongly with Chinese strain 84-7 within clade 2, more specifically cluster 2a. Signature sequences for the cluster 2a were identified and found to play an important role in the separation of cluster 2a strains from other clade 2 strains, as shown in substitution studies. Further genetic analysis with additional strains isolated from Japan, China, and other Asian countries would provide a novel insight into the significance of two distinct subclades within clade 2.

  3. Antagonistic bioactivity of endophytic strains isolated from Salvia ...

    African Journals Online (AJOL)

    The antibiotic-producing potential of endophytic populations from medical plant of Salvia miltiorrhiza was examined. A total of 63 isolates was screened against five fungal and three bacterial species for the production of antimicrobial compounds. It showed that more isolates was antagonistic to fungi than to bacteria.

  4. Molecular characterization of indigenous Bacillus thuringiensis strains isolated from Kashmir valley

    National Research Council Canada - National Science Library

    Reyaz, A L; Gunapriya, L; Indra Arulselvi, P

    2017-01-01

    Bacillus thuringiensis (Bt) being an eco-friendly bioinsecticide is effectively used in pest management strategies and, therefore, isolation and identification of new strains effective against a broad range of target pests is important...

  5. Complete Genome Sequence of a Dengue Virus Serotype 4 Strain Isolated in Roraima, Brazil

    Science.gov (United States)

    Souza, Victor C.; Silva, George A. V.; Maito, Rodrigo M.; Granja, Fabiana; Siqueira, Thalita; Acosta, Pablo O. A.

    2012-01-01

    Dengue is the most important arboviral disease worldwide. We report the complete genome sequence of a dengue virus serotype 4, genotype II strain isolated in 2010 from a patient with classical dengue fever in Boa Vista, Roraima, Brazil. PMID:22247521

  6. Genetic diversity of Mycobacterium tuberculosis strains isolated in Algeria: Results of spoligotyping

    Directory of Open Access Journals (Sweden)

    Malika Ifticene

    2015-01-01

    Conclusion: The study of strains isolated between January and December 2011 has allowed insight into the frequency of different genomic families and the importance of existing clusters in the population of central and eastern Algeria.

  7. Genotypic diversity and pathogenic potential of Yersinia enterocolitica biotype 2 strains isolated in Brazil.

    Science.gov (United States)

    Frazão, M R; Falcão, J P

    2015-04-01

    To investigate the pathogenic potential and genotypic diversity of Yersinia enterocolitica biotype 2 strains isolated in Brazil and to compare these strains with other Y. enterocolitica biotypes using ERIC-PCR and PFGE. Forty strains of Y. enterocolitica biotype 2 (B2) isolated from humans (5), the environment (34) and animal (1), in Brazil over 19 years were studied. In addition to these isolates, we also analysed 26 Y. enterocolitica strains belonging to the biotypes 1A, 1B, and 3-5. All of the B2 strains contained the genes inv, ail, ystA, hreP, tccC and myfA. The genes fepD and fes were detected in 39 (97·5%) strains, virF was found in three (7·5%) strains, and ystB and fepA were not detected in any strains. The B2 strains showed genotypic similarities of more than 84·8% by ERIC-PCR and of more than 69·0% by PFGE. The pathogenic potential of the B2 strains examined in this study was highlighted by the occurrence of the majority of the virulence markers searched. The results of the ERIC-PCR and PFGE showed that the B2 strains evaluated in this study had a high genotypic similarity, suggesting that these strains differed little over the 19 year study period and that the environment was a possible source of contamination of humans and animals in Brazil. Furthermore, the ERIC-PCR technique grouped the strains belonging to Y. enterocolitica biotypes 1A, 1B, 2, 3, 4 and 5 according to their pathogenicity. This study provided new information about the pathogenic potential and genotypic similarity of Y. enterocolitica B2 isolated from diverse sources in Brazil. Furthermore, ERIC-PCR showed to be a valuable tool for grouping Y. enterocolitica of different biotypes according their pathogenicity. © 2015 The Society for Applied Microbiology.

  8. Draft genome sequences of five clinical Enterococcus cecorum strains isolated from different poultry species in Poland

    DEFF Research Database (Denmark)

    Dolka, Beata; Olsen, Rikke Heidemann; Thøfner, Ida

    2015-01-01

    Here, we report five draft genome sequences of Enterococcus cecorum strains that were isolated from different bird species of affected poultry flocks (commercial broilers [CB], broiler breeders [BB], commercial layers [CL], ducks [D], and geese [G]) in Poland.......Here, we report five draft genome sequences of Enterococcus cecorum strains that were isolated from different bird species of affected poultry flocks (commercial broilers [CB], broiler breeders [BB], commercial layers [CL], ducks [D], and geese [G]) in Poland....

  9. Isolation and Fatty Acid Profile of Selected Microalgae Strains from the Red Sea for Biofuel Production

    OpenAIRE

    Hugo Pereira; Luísa Barreira; Luísa Custódio; Salman Alrokayan; Fouzi Mouffouk; João Varela; Khalid M. Abu-Salah; Radhouan Ben-Hamadou

    2013-01-01

    The isolation of lipid-rich autochthonous strains of microalgae is a crucial stage for the development of a microalgae-based biofuel production plant, as these microalgae already have the necessary adaptations to withstand competition, predation and the temperatures observed at each production site. This is particularly important in extreme climates such as in Saudi Arabia. Resorting to fluorescence activated cell sorting (FACS) we screened for and isolated several microalgal strains from sam...

  10. RAPD Analysis and Antibiotic Susceptibility for Mycobacterium tuberculosis Strains Isolated from Different Locations in Egypt

    Directory of Open Access Journals (Sweden)

    Ali, A. M.

    2011-01-01

    Full Text Available The routine identification of mycobacterial strains isolated from patients in different locations in Egypt was confirmed by specific DNA fragment amplification. The susceptibilities of 72 Mycobacterium tuberculosis strains against the four antibiotics used in tuberculosis treatment (Isoniazid, INH; Rifampicin, Rif; Streptomycin, St and Ethambutol, E were examined. Our results indicated that, multi drug resistant tuberculosis (MDR-TB represents about 19.5% of the tested strains, whereas sensitive strains represented 26.4%. The genetic polymorphism of the tested strains was examined using RAPD analysis. Six selected strains represent the different antibiotic susceptibility groups were examined using RAPD fingerprinting. No difference between the strains was recorded using the RFLP analysis of amplified specific fragment. The discrimination power of RAPD analysis was inadequate to clarify the genetic correlation between the tested strains. MDR-TB was approximately double time in 2008 compared with the value in 2007. Most of the new MDRTB was correlated with resident dense population regions.

  11. Cross-Comparison of Leaching Strains Isolated from Two Different Regions: Chambishi and Dexing Copper Mines

    Directory of Open Access Journals (Sweden)

    Baba Ngom

    2014-01-01

    Full Text Available A cross-comparison of six strains isolated from two different regions, Chambishi copper mine (Zambia, Africa and Dexing copper mine (China, Asia, was conducted to study the leaching efficiency of low grade copper ores. The strains belong to the three major species often encountered in bioleaching of copper sulfide ores under mesophilic conditions: Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, and Leptospirillum ferriphilum. Prior to their study in bioleaching, the different strains were characterized and compared at physiological level. The results revealed that, except for copper tolerance, strains within species presented almost similar physiological traits with slight advantages of Chambishi strains. However, in terms of leaching efficiency, native strains always achieved higher cell density and greater iron and copper extraction rates than the foreign microorganisms. In addition, microbial community analysis revealed that the different mixed cultures shared almost the same profile, and At. ferrooxidans strains always outcompeted the other strains.

  12. Primary isolation strain determines both phage type and receptors recognised by Campylobacter jejuni bacteriophages.

    Directory of Open Access Journals (Sweden)

    Martine C Holst Sørensen

    Full Text Available In this study we isolated novel bacteriophages, infecting the zoonotic bacterium Campylobacter jejuni. These phages may be used in phage therapy of C. jejuni colonized poultry to prevent spreading of the bacteria to meat products causing disease in humans. Many C. jejuni phages have been isolated using NCTC12662 as the indicator strain, which may have biased the selection of phages. A large group of C. jejuni phages rely on the highly diverse capsular polysaccharide (CPS for infection and recent work identified the O-methyl phosphoramidate modification (MeOPN of CPS as a phage receptor. We therefore chose seven C. jejuni strains each expressing different CPS structures as indicator strains in a large screening for phages in samples collected from free-range poultry farms. Forty-three phages were isolated using C. jejuni NCTC12658, NCTC12662 and RM1221 as host strains and 20 distinct phages were identified based on host range analysis and genome restriction profiles. Most phages were isolated using C. jejuni strains NCTC12662 and RM1221 and interestingly phage genome size (140 kb vs. 190 kb, host range and morphological appearance correlated with the isolation strain. Thus, according to C. jejuni phage grouping, NCTC12662 and NCTC12658 selected for CP81-type phages, while RM1221 selected for CP220-type phages. Furthermore, using acapsular ∆kpsM mutants we demonstrated that phages isolated on NCTC12658 and NCTC12662 were dependent on the capsule for infection. In contrast, CP220-type phages isolated on RM1221 were unable to infect non-motile ∆motA mutants, hence requiring motility for successful infection. Hence, the primary phage isolation strain determines both phage type (CP81 or CP220 as well as receptors (CPS or flagella recognised by the isolated phages.

  13. A rhizobia strain isolated from root nodule of gymnosperm Podocarpus macrophyllus.

    Science.gov (United States)

    Huang, BaoLing; Lü, ChengQun; Wu, Bo; Fan, LiQin

    2007-04-01

    Symbiotic nitrogen fixation of rhizobia and leguminous plants is considered as the most important biologic nitrogen fixation system on earth. Symbiotic nodulation of gymnosperm Podocarpus macrophyllus and rhizobia has never been reported. In this study, 11 endophytic bacteria strains were isolated from root nodules of P. macrophyllus and its variation P. macrophyllus var. maki. The plant infection tests on these strains indicated that the isolated strains could be nodulated on P. macrophyllus plants, and weak nitrogenase activity of nodules was found in acetylene reduction method. According to the physiological and biochemical characteristics of the 11 strains, GXLO 02 was selected as the representative strain. 16S rDNA full-length sequence analysis of GXLO 02 confirmed that the representative strain GXLO 02 belongs to Rhizobium sp.

  14. Phenotypic and molecular characterization of Salmonella Enteritidis strains isolated in São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Fernandes Sueli A.

    2003-01-01

    Full Text Available In São Paulo State, Brazil, the epidemic increase in isolation of Salmonella Enteritidis has been observed since 1994. A total of 105 S. Enteritidis strains (72 from human and 33 from non-human sources isolated during the period 1975-1995, previously characterized by phage typing, was analyzed by antimicrobial susceptibility, plasmid profile, and ribotyping. Over 70% of the strains were susceptible to all antimicrobial agents tested, however, multiple resistance to antimicrobials was observed among the studied strains, mainly those from hospitalized patients. Phage type 8 (PT-8 was predominant among the strains isolated during the period of 1975-1992, but in the following years, PT-4 was the most frequent phage type identified. Seven different plasmid profiles were detected and 96% of the isolates harbored a plasmid of approximately 36 MDa. Ribotyping discriminated fourteen ribotypes (R1 to R14 among the strains examined. By analysis of dendrogram the strains were included in three groups with similarity level of 60%. The obtained results indicate that, a single ribotype (R11, determined for PT-4 strains isolated from 1993, characterizes the epidemic clone of S. Enteritidis in our region.

  15. Variability of isolated colonies in bean nodulating Rhizobium strains before and after exposure to high temperature

    Directory of Open Access Journals (Sweden)

    Raposeiras Rui

    2002-01-01

    Full Text Available Irregular response to bean plants to Rhizobium inoculation has been attributed to among other factors, low competitive ability, low N2 fixation efficiency and genetic instability of the symbiont. This genetic instability caused by high rates of genomic rearrangements and/or plasmid deletions can be accentuated by high temperatures. This fact may limit the utilization of these strains as inoculants, especially in tropical soils. In this study, the variability of isolated colonies derived from effective R. leguminosarum bv. phaseoli (SLP1.3 and BR 10.026 and R tropici (SLA2.2 and BR322 strains was evaluated before and after exposure to high temperatures (four consecutive thermal shocks at 45masculineC. This evaluation involved plant dry matter analysis of inoculated plants and genotypic (plasmid profile and genomic patterns via RAPD analysis of the Rhizobium strains. The results evidenced that high temperature improve the natural performance variability especially between isolated colonies from R. leguminosarum bv. phaseoli strains. The plasmid profile of isolated colonies from R. tropici strains were identical regardless of temperature treatment whereas isolated colonies from R. leguminosarum bv. phaseoli alterations were detected especially after the thermal treatment. The genomic patterns generated by AP-PCR showed more alterations and genetic variation in isolated colonies from R. leguminosarum bv. phaseoli strains indicating that R. tropici strains are more stable and lower affected by high temperature.

  16. Variable characteristics of bacteriocin-producing Streptococcus salivarius strains isolated from Malaysian subjects.

    Science.gov (United States)

    Barbour, Abdelahhad; Philip, Koshy

    2014-01-01

    Salivaricins are bacteriocins produced by Streptococcus salivarius, some strains of which can have significant probiotic effects. S. salivarius strains were isolated from Malaysian subjects showing variable antimicrobial activity, metabolic profile, antibiotic susceptibility and lantibiotic production. In this study we report new S. salivarius strains isolated from Malaysian subjects with potential as probiotics. Safety assessment of these strains included their antibiotic susceptibility and metabolic profiles. Genome sequencing using Illumina's MiSeq system was performed for both strains NU10 and YU10 and demonstrating the absence of any known streptococcal virulence determinants indicating that these strains are safe for subsequent use as probiotics. Strain NU10 was found to harbour genes encoding salivaricins A and 9 while strain YU10 was shown to harbour genes encoding salivaricins A3, G32, streptin and slnA1 lantibiotic-like protein. Strain GT2 was shown to harbour genes encoding a large non-lantibiotic bacteriocin (salivaricin-MPS). A new medium for maximum biomass production buffered with 2-(N-morpholino)ethanesulfonic acid (MES) was developed and showed better biomass accumulation compared with other commercial media. Furthermore, we extracted and purified salivaricin 9 (by strain NU10) and salivaricin G32 (by strain YU10) from S. salivarius cells grown aerobically in this medium. In addition to bacteriocin production, S. salivarius strains produced levan-sucrase which was detected by a specific ESI-LC-MS/MS method which indicates additional health benefits from the developed strains. The current study established the bacteriocin, levan-sucrase production and basic safety features of S. salivarius strains isolated from healthy Malaysian subjects demonstrating their potential for use as probiotics. A new bacteriocin-production medium was developed with potential scale up application for pharmaceuticals and probiotics from S. salivarius generating different

  17. Variable characteristics of bacteriocin-producing Streptococcus salivarius strains isolated from Malaysian subjects.

    Directory of Open Access Journals (Sweden)

    Abdelahhad Barbour

    Full Text Available BACKGROUND: Salivaricins are bacteriocins produced by Streptococcus salivarius, some strains of which can have significant probiotic effects. S. salivarius strains were isolated from Malaysian subjects showing variable antimicrobial activity, metabolic profile, antibiotic susceptibility and lantibiotic production. METHODOLOGY/PRINCIPAL FINDINGS: In this study we report new S. salivarius strains isolated from Malaysian subjects with potential as probiotics. Safety assessment of these strains included their antibiotic susceptibility and metabolic profiles. Genome sequencing using Illumina's MiSeq system was performed for both strains NU10 and YU10 and demonstrating the absence of any known streptococcal virulence determinants indicating that these strains are safe for subsequent use as probiotics. Strain NU10 was found to harbour genes encoding salivaricins A and 9 while strain YU10 was shown to harbour genes encoding salivaricins A3, G32, streptin and slnA1 lantibiotic-like protein. Strain GT2 was shown to harbour genes encoding a large non-lantibiotic bacteriocin (salivaricin-MPS. A new medium for maximum biomass production buffered with 2-(N-morpholinoethanesulfonic acid (MES was developed and showed better biomass accumulation compared with other commercial media. Furthermore, we extracted and purified salivaricin 9 (by strain NU10 and salivaricin G32 (by strain YU10 from S. salivarius cells grown aerobically in this medium. In addition to bacteriocin production, S. salivarius strains produced levan-sucrase which was detected by a specific ESI-LC-MS/MS method which indicates additional health benefits from the developed strains. CONCLUSION: The current study established the bacteriocin, levan-sucrase production and basic safety features of S. salivarius strains isolated from healthy Malaysian subjects demonstrating their potential for use as probiotics. A new bacteriocin-production medium was developed with potential scale up application for

  18. Heavy metal resistance by two bacteria strains isolated from a ...

    African Journals Online (AJOL)

    USER

    2010-06-28

    T3-03, were isolated from the biggest tailing in Asia-Dexing ... well as affect the qualitative and quantitative structure of microbial communities (Giller et al., .... The enriched bacterial cells were collected. Olympus CX 31 optical.

  19. Isolation and characterization of a new Pseudomonas- related strain ...

    African Journals Online (AJOL)

    SAM

    2014-04-02

    . 64(11):4396-4402. Wei GH, Yu JF, Zhu YH, Chen WM, Wang L (2008). Characterization of phenol degradation by Rhizobium sp CCNWTB 701 isolated from. Astragalus chrysopteru in mining tailing region. J. Hazard. Mater.

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

    Directory of Open Access Journals (Sweden)

    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

  1. Isolation and characterization of some dominant yeast strains for ...

    African Journals Online (AJOL)

    The isolate ACP12 showed even higher population number (9.7 ± 1.00 x 107 CFU/ml) than the standard culture (Saccharomyces cerevisiae) with colony count of 8.7 ± 1.00 x 107 CFU/ml at 30°C and at pH 5. Based on morphological, physiological and biochemical characteristics, the two isolates (ACE12 and ACP12) were ...

  2. Draft Genome Sequence of Micromonospora sp. Strain HK10, Isolated from Kaziranga National Park, India

    Science.gov (United States)

    Talukdar, Madhumita; Das, Dhrubajyoti; Borah, Chiranjeeta; Deka Boruah, Hari Prasanna; Bora, Tarun Chandra

    2016-01-01

    We report the 6.92-Mbp genome sequence of Micromonospora sp. HK10, isolated from soil samples collected from Kaziranga National Park, Assam, India. The full genome of strain Micromonospora sp. strain HK10 consists of 6,911,179 bp with 73.39% GC content, 6,196 protein-coding genes, and 86 RNAs. PMID:27516496

  3. Isolation of a nitrate-reducing bacteria strain from oil field brine and ...

    African Journals Online (AJOL)

    A nitrate-reducing bacteria (NRB) strain with vigorous growth, strong nitrate reduction ability, strain B9 2-1, was isolated from Suizhong36-1 oilfield, its routine identification and analysis of 16S rRNA and also the competitive inhibition experiments with the enrichment of sulfate-reducing bacteria (SRB) were carried out.

  4. Complete Genome Sequence of Pediococcus pentosaceus Strain wikim 20, Isolated from Korean Kimchi

    Science.gov (United States)

    Lee, Se Hee; Jung, Min Young; Park, Boyeon; Sung-Oh, Sohn; Park, Hae Woong; Choi, Hak-Jong

    2016-01-01

    Pediococcus pentosaceus strain wikim 20 is a lactic acid bacterium that was isolated from kimchi, a representative traditional Korean fermented food. Here, we announce the complete genome sequence of P. pentosaceus strain wikim 20 consisting of a 1,830,629-bp chromosome and provide a description of its annotation. PMID:27834699

  5. Complete Genome Sequence of Lactobacillus curvatus Strain WiKim38 Isolated from Kimchi

    Science.gov (United States)

    Lee, Se Hee; Jung, Min Young; Song, Jung-Hee; Lee, Moeun

    2017-01-01

    ABSTRACT Lactobacillus curvatus WiKim38 is a potential probiotic strain isolated from kimchi, a traditional Korean fermented food. The complete genome of the WiKim38 strain consisted of a circular chromosome of 1,940,170 bp in length with a G+C content of 41.93%. PMID:28473381

  6. Isolation of a nitrate-reducing bacteria strain from oil field brine and ...

    African Journals Online (AJOL)

    Jane

    2011-08-31

    Aug 31, 2011 ... A nitrate-reducing bacteria (NRB) strain with vigorous growth, strong nitrate reduction ability, strain B9. 2-1, was isolated from Suizhong36-1 oilfield, its routine identification and analysis of 16S rRNA and also the competitive inhibition experiments with the enrichment of sulfate-reducing bacteria (SRB) were.

  7. Genome sequence of Janthinobacterium sp. strain PAMC 25724, isolated from alpine glacier cryoconite.

    Science.gov (United States)

    Kim, Su Jin; Shin, Seung Chul; Hong, Soon Gyu; Lee, Yung Mi; Lee, Hyoungseok; Lee, Jungeun; Choi, In-Geol; Park, Hyun

    2012-04-01

    The draft genome of Janthinobacterium sp. strain PAMC 25724, which is a violacein-producing psychrotolerant bacterium, was determined. The strain was isolated from glacier cryoconite of the Alps mountain permafrost region. The sequence will allow identification and characterization of the genetic determination of its cold-adaptive properties.

  8. Isolation and characterization of the E. coli membrane protein production strain Mutant56(DE3)

    DEFF Research Database (Denmark)

    Baumgarten, Thomas; Schlegel, Susan; Wagner, Samuel

    2017-01-01

    Membrane protein production is usually toxic to E. coli. However, using genetic screens strains can be isolated in which the toxicity of membrane protein production is reduced, thereby improving production yields. Best known examples are the C41(DE3) and C43(DE3) strains, which are both derived...

  9. Isolation and characterization of the E. coli membrane protein production strain Mutant56(DE3)

    NARCIS (Netherlands)

    Baumgarten, Thomas; Schlegel, Susan; Wagner, Samuel; Löw, Mirjam; Eriksson, Jonas; Bonde, Ida; Herrgård, Markus J; Heipieper, Hermann J; Nørholm, Morten H H; Slotboom, Dirk Jan; de Gier, Jan-Willem

    2017-01-01

    Membrane protein production is usually toxic to E. coli. However, using genetic screens strains can be isolated in which the toxicity of membrane protein production is reduced, thereby improving production yields. Best known examples are the C41(DE3) and C43(DE3) strains, which are both derived from

  10. DIFFERENTIATION OF BIOMASS COMPOSITION BETWEEN ISOLATED AND COMMERCIAL STRAINS OF MICROALGAE

    Directory of Open Access Journals (Sweden)

    ZURAIFAH BT. MINHAT

    2016-05-01

    Full Text Available Biomass produced from the cultivation of microalgae has many potential in producing high value product. Main objective of this research is to investigate the potential of biomass produced by algae from isolated strains and commercialize strains as well as to compare their potential in producing high quality of biomass. Isolated microalgaestrains used were C. sorokiana (UKM 3 and Characium sp. (UKM 1 extracted from Palm Oil Mill Effluent (POME and commercial strains used were C. vulgaris and Ankistrodesmus sp.. Results shows that the commercial strain Ankistrodesmus sp has a highest biomass’s weight which is 3.0 g/L, followed by another type of commercial strain, C. vulgaris which is at 2.72g/L and the other two types of isolated strains, C.sorokiniana and Characium sp. both at 2.52g/L and 2.27g/L respectively. For the biomass composition, the highest concentration of protein was found in locally isolated strain, Characium sp. with concentration of 0.52g/L, while for carbohydrates and lipid were found highest in commercial strain, C. vulgaris with concentration at 0.27g/L and 2.07g/L respectively.

  11. Genome Sequence of Lawsonia intracellularis Strain N343, Isolated from a Sow with Hemorrhagic Proliferative Enteropathy.

    Science.gov (United States)

    Sait, Michelle; Aitchison, Kevin; Wheelhouse, Nick; Wilson, Kim; Lainson, F Alex; Longbottom, David; Smith, David G E

    2013-01-01

    Lawsonia intracellularis is the etiological agent of proliferative enteropathy (PE), causing mild or acute hemorrhagic diarrhea in infected animals. Here we report the genome sequence of strain N343, isolated from a sow that died of hemorrhagic PE. N343 contains 24 single nucleotide polymorphisms and 90 indels compared to the reference strain PHE/MN1-00.

  12. Increasing multidrug resistance in Helicobacter pylori strains isolated from children and adults in Mexico.

    Science.gov (United States)

    Torres, J; Camorlinga-Ponce, M; Pérez-Pérez, G; Madrazo-De la Garza, A; Dehesa, M; González-Valencia, G; Muñoz, O

    2001-07-01

    The susceptibilities to three antimicrobials of 195 Helicobacter pylori strains isolated from Mexican patients is reported; 80% of the strains were resistant to metronidazole, 24% were resistant to clarithromycin, and 18% presented a transient resistance to amoxicillin. Resistance to two or more antimicrobials increased significantly from 1995 to 1997.

  13. FERMENTATION OF INULIN BY A NEW STRAIN OF CLOSTRIDIUM-THERMOAUTOTROPHICUM ISOLATED FROM DAHLIA TUBERS

    NARCIS (Netherlands)

    DRENT, WJ; GOTTSCHAL, JC

    1991-01-01

    A new inulin-fermenting strain of Clostridium thermoautotrophicum was isolated through enrichment on dahlia tubers, and subsequent plating on agar media with undissolved inulin. Both the cell-bound and cell-free inulinase(s) functioned optimally at 60-degrees-C and at neutral pH. This new strain I1

  14. Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood

    Directory of Open Access Journals (Sweden)

    Kirill V. Sergueev

    2017-06-01

    Full Text Available For decades, bacteriophages (phages have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types.

  15. 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.

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

    Science.gov (United States)

    Allen, Adrian; Breadon, Eleanor; Byrne, Andrew; Mallon, Thomas; Skuce, Robin; Groussaud, Pauline; Dainty, Amanda; Graham, Judith; Jones, Kerri; Pollock, Lorraine; Whatmore, Adrian

    2015-01-01

    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.

  17. Molecular Epidemiology of Brucella abortus in Northern Ireland—1991 to 2012

    Science.gov (United States)

    Byrne, Andrew; Mallon, Thomas; Skuce, Robin; Groussaud, Pauline; Dainty, Amanda; Graham, Judith; Jones, Kerri; Pollock, Lorraine; Whatmore, Adrian

    2015-01-01

    Background 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. Results 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. Conclusions 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. PMID:26325586

  18. Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood

    Science.gov (United States)

    Sergueev, Kirill V.; Filippov, Andrey A.; Nikolich, Mikeljon P.

    2017-01-01

    For decades, bacteriophages (phages) have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter) within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types. PMID:28604602

  19. Virulence genes of Escherichia coli strains isolated from mastitic milk.

    Science.gov (United States)

    Bean, A; Williamson, J; Cursons, R T

    2004-08-01

    Escherichia coli, a Gram-negative environmental pathogen associated with bovine mastitis was isolated from the milk of 34 symptomatic cows that had been diagnosed with clinical mastitis. Eighty isolates were obtained over a 17-month period and these isolates were screened by DNA amplification for the following E. coli virulence genes: cnf1, cnf2, eaeA, eagg, einv, ltx1, stx1, stx2 and vt2e. Thirty of the bacterial isolates, obtained from 23 different cows, had toxin genes identified in their DNA. The most common virulence gene detected was stx1, with a prevalence of 31%, followed by cnf2 (7.5%), vt2e (6.25%) and eaeA (4%). The possession of different virulence genes by the bacterial isolates had no discernable impact on the health status of the cows as there was no correlation between the potential for toxin production by the E. coli isolates and the systemic clinical condition of the respective infected cows.

  20. 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

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

    Science.gov (United States)

    Li, Xinna; He, Yongqun

    2012-01-01

    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 presentation, and T

  2. Isolation and characterization of a fluoranthene-utilizing strain of Pseudomonas paucimobilis.

    OpenAIRE

    Mueller, J G; Chapman, P J; Blattmann, B O; Pritchard, P. H.

    1990-01-01

    A soil bacterium capable of utilizing fluoranthene as the sole source of carbon and energy for growth was purified from a seven-member bacterial community previously isolated from a creosote waste site for its ability to degrade polycyclic aromatic hydrocarbons. By standard bacteriological methods, this bacterium was characterized taxonomically as a strain of Pseudomonas paucimobilis and was designated strain EPA505. Utilization of fluoranthene by strain EPA 505 was demonstrated by increase i...

  3. Technological Aptitude and Applications of Leuconostoc mesenteroides Bioactive Strains Isolated from Algerian Raw Camel Milk

    OpenAIRE

    Zineb Benmechernene; Hanane Fatma Chentouf; Bellil Yahia; Ghazi Fatima; Marcos Quintela-Baluja; Pilar Calo-Mata; Jorge Barros-Velázquez

    2013-01-01

    Two strains (B7 and Z8) of the Leuconostoc mesenteroides subspecies mesenteroides that were isolated from Algerian camel milk from an initial pool of 13 strains and demonstrated a high ability to inhibit the growth of Listeria spp. were selected and characterised at the phenotypic and genotypic levels. Probiotic profiling and inhibition spectra against food borne pathogens in mixed cultures were also investigated. The bacteriocin produced by L. mesenteroides strain B7 was identified as le...

  4. Isolation, cultivation and molecular characterization of a new Trypanosoma equiperdum strain in Mongolia

    OpenAIRE

    Suganuma, Keisuke; Narantsatsral, Sandagdorj; Battur, Banzragch; Yamasaki, Shino; Otgonsuren, Davaajav; Musinguzi, Simon Peter; Davaasuren, Batdorj; Battsetseg, Badgar; Inoue, Noboru

    2016-01-01

    Background Trypanosoma equiperdum causes dourine via sexual transmission in Equidae. T. equiperdum is classified under the subgenus Trypanozoon along with the T. brucei sspp. and T. evansi; however, the species classification of Trypanozoon remains a controversial topic due to the limited number of T. equiperdum reference strains. In addition, it is possible that some were misclassified T. evansi strains. Thus, there is a strong need for a new T. equiperdum strain directly isolated from the g...

  5. Characterization of functional properties of Enterococcus faecium strains isolated from human gut.

    Science.gov (United States)

    İspirli, Hümeyra; Demirbaş, Fatmanur; Dertli, Enes

    2015-11-01

    The aim of this work was to characterize the functional properties of Enterococcus faecium strains identified after isolation from human faeces. Of these isolates, strain R13 showed the best resistance to low pH, bile salts, and survival in the simulated in vitro digestion assay, and demonstrated an important level of adhesion to hexadecane as a potential probiotic candidate. Analysis of the antibiotic resistance of E. faecium strains indicated that in general these isolates were sensitive to the tested antibiotics and no strain appeared to be resistant to vancomycin. Examination of the virulence determinants for E. faecium strains demonstrated that all strains contained the virulence genes common in gut- and food-originated enterococci, and strain R13 harboured the lowest number of virulence genes. Additionally, no strain contained the genes related to cytolysin metabolism and showed hemolytic activity. The antimicrobial role of E. faecium strains was tested against several pathogens, in which different levels of inhibitory effects were observed, and strain R13 was inhibitory to all tested pathogens. PCR screening of genes encoding enterocin A and B indicated the presence of these genes in E. faecium strains. Preliminary characterization of bacteriocins revealed that their activity was lost after proteolytic enzyme treatments, but no alteration in antimicrobial activity was observed at different pHs (3.5 to 9.5) and after heat treatments. In conclusion, this study revealed the functional characteristics of E. faecium R13 as a gut isolate, and this strain could be developed as a new probiotic after further tests.

  6. Antagonistic Activity of Three Newly Isolated Yeast Strains from the Surface of Fruits

    Directory of Open Access Journals (Sweden)

    Ortansa Csutak

    2013-01-01

    Full Text Available Three Metschnikowia pulcherrima strains isolated from white grapes(strains designated SG1 and SG2 and cherries (strain CPM1 from Romania were tested for antagonistic activity against natural and reference yeasts, fungi and E. coli. The best results were obtained for SG2 strain against the following microbes isolated from natural environments: yeasts Candida tropicalis OT4 and Candida albicans Cc, and fungal strains Botrytis cinerea BCS and BCF1. Using 2 % sodium hydrogen carbonate and 1 or 2 % calcium chloride, antimicrobial activities of strains SG1 and SG2 were maintained or even enhanced, while strain CPM1 was less influenced. Antimicrobial tests revealed four types of colonies and halos of strains SG1, SG2 and CPM1 reflecting differences in iron immobilization under similar growth conditions. Antifungal activity and the level of pigment production increased at 25 °C, and the largest diffusible pigment area was obtained by strain SG2, which altered Botrytis conidia germination and induced modifications of mycelium. Competition for iron seems to be the major mechanism of antagonistic action of our strains, which is reduced/absent under iron chloride addition. Lowering of antifungal activity by heating or proteolytic treatment indicates that a protein might also be involved. The inhibitory activity of yeast supernatants was less important, suggesting a cell-related mechanism. When combined, strains SG1 and SG2 had synergistic activity, which was reduced in the presence of CPM1 due to possible production of an inhibitory compound.

  7. CHARACTERIZATION OF EXTENDED-SPECTRUM Β-LACTAMASE-PRODUCING ESCHERICHIA COLI STRAINS ISOLATED FROM DAIRY PRODUCTS

    Directory of Open Access Journals (Sweden)

    Rahem Khoshbakht

    2014-02-01

    Full Text Available Extended-spectrum β-lactamases (ESBLs are enzymes that hydrolyze the β-lactam ring, and ESBL-producing E. coli has rapidly spread worldwide with pose a serious hazard for humans. The aim of this study was to determine the prevalence of ESBL producing E. coli and molecular evaluation of four ESBL-associated genes among E. coli strains isolated from milk and cheese in southern Iran. Antibiotic susceptibility test was carried out for a total of 150 isolates of E. coli, previously collected from dairy products. ESBL production was screened using a double-disc synergy test (DDST and presence of four ESBL genes (PER, VEB, TEM and CTX-M was tested using PCR. Among 150 E. coli strains 57 (38% isolates were identified as ESBL-producing strains. All ESBL positive isolates could be typed for one or more genes and the most prevalent ESBL-associated gene was CTX-M (80.7%. The PER gene was not present among isolates. Isolates showed high susceptibility to imipe¬nem and cefoxitin. The results showed the high prevalence of ESBL producing E. coli strains among dairy products and high occurrence of CTX-M-associated ESBL activity among isolates indicating the hazards of increasing the strains with antibiotic resistance which can transfer to human trough the dairy food products.

  8. Isolating and evaluating lactic acid bacteria strains for effectiveness of Leymus chinensis silage fermentation.

    Science.gov (United States)

    Zhang, Q; Li, X J; Zhao, M M; Yu, Z

    2014-10-01

    Five LAB strains were evaluated using the acid production ability test, morphological observation, Gram staining, physiological, biochemical and acid tolerance tests. All five strains (LP1, LP2, LP3, LC1 and LC2) grew at pH 4·0, and LP1 grew at 15°C. Strains LP1, LP2 and LP3 were identified as Lactobacillus plantarum, whereas LC1 and LC2 were classified as Lactobacillus casei by sequencing 16S rDNA. The five isolated strains and two commercial inoculants (PS and CL) were added to native grass and Leymus chinensis (Trin.) Tzvel. for ensiling. All five isolated strains decreased the pH and ammonia nitrogen content, increased the lactic acid content and LP1, LP2 and LP3 increased the acetic content and lactic/acetic acid ratio of L. chinensis silage significantly. The five isolated strains and two commercial inoculants decreased the butyric acid content of the native grass silage. LP2 treatment had lower butyric acid content and ammonia nitrogen content than the other treatments. The five isolated strains improved the quality of L. chinensis silage. The five isolated strains and the two commercial inoculants were not effective in improving the fermentation quality of the native grass silage, but LP2 performed better comparatively. Significance and impact of the study: Leymus chinensis is an important grass in China and Russia, being the primary grass of the short grassland 'steppe' regions of central Asia. However, it has been difficult to make high-quality silage of this species because of low concentration of water-soluble carbohydrates (WSC). Isolating and evaluating lactic acid bacteria strains will be helpful for improving the silage quality of this extensively grown species. © 2014 The Society for Applied Microbiology.

  9. Isolation of soil thermophilic strains of actinomycetes for the ...

    African Journals Online (AJOL)

    amylase enzyme produced by the strains. Extraction of α-amylase was done in liquid medium. The assay was observed by measuring the release of reducing sugar (RS) by 3,5- dinitrosalicylic acid (DNS) method and expressed in International ...

  10. methoxyethanol by a new bacterium isolate Pseudomonas sp. Strain ...

    African Journals Online (AJOL)

    Michael Horsfall

    at room temperature by using a Clark - Type electrode Model 10 Ranke Brothers, Birmingham,. Great Britain) ... strain VB was determined with Clarke-Type electrode. Briefly, 3ml of 50mM phosphate buffer. (pH 7.2) was ... Aerobic transformation of 2 – methoxyethanol and hypothetical pathways intermediates: Studies of the.

  11. Isolation and characterization of a new Pseudomonas- related strain ...

    African Journals Online (AJOL)

    SAM

    2014-04-02

    Apr 2, 2014 ... bacterial strains from phenol-formaldehyde resin manufacturing industrial wastewater. J. Hazard. Mater. B127:238-243. Ausubel FM , Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA,. Struhl K(1995). Current Protocols in Molecular Biology. Greene. Publishing Associates/Wiley Interscience, New ...

  12. Isolation of soil thermophilic strains of actinomycetes for the ...

    African Journals Online (AJOL)

    use

    2011-12-05

    Dec 5, 2011 ... proportional to the amount of α-amylase enzyme produced by the strains. Extraction of α-amylase was done in liquid medium. The assay was observed by measuring the release of reducing sugar (RS) by 3,5- dinitrosalicylic acid (DNS) method and expressed in International Units (IU). The thermophilic ...

  13. Isolation and Identification of a Brevibacterium linens strain ...

    African Journals Online (AJOL)

    It showed tyrosine clearing and had meso-DAP as the characteristic cell wall amino acid. On the basis of the morphological, physiological, and biochemical tests the organism was identified as Brevibacterium linens. To our knowledge, this is the first report of any Brevibacterium strain able to degrade PNP. African Journal of ...

  14. Ethanol production potential of local yeast strains isolated from ripe ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-16

    May 16, 2008 ... has been Saccharomyces cerevisiae. This yeast also has the ability to produce ethanol which is not contaminated by other products from the substrate. Banana peels are readily available agricultural waste in. Nigeria, yet they seem to be underutilized as potential growth medium for local yeast strains, ...

  15. Characterization of Streptomyces strain SLO-105 isolated from Lake ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-16

    Nov 16, 2009 ... Hopwood et al., 1985). Morphological characteristics. The micro morphology of the strain was carried out using a scan- ning electron microscope Philips XL 30 ESEM-FEG (Philips Optique. Electronique, Limeil Brévannes, FRANCE). The sample used for observation has been cultured for 14 days on Yeast ...

  16. Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil

    Directory of Open Access Journals (Sweden)

    Carolina S.A.B. Santos

    2012-11-01

    Full Text Available Few data are available on the prevalence and risk factors of Chlamydophila abortus infection in goats in Brazil. A cross-sectional study was carried out to determine the flock-level prevalence of C. abortus infection in goats from the semiarid region of the Paraíba State, Northeast region of Brazil, as well as to identify risk factors associated with the infection. Flocks were randomly selected and a pre-established number of female goats > 12 mo old were sampled in each of these flocks. A total of 975 serum samples from 110 flocks were collected, and structured questionnaire focusing on risk factors for C. abortus infection was given to each farmer at the time of blood collection. For the serological diagnosis the complement fixation test (CFT using C. abortus S26/3 strain as antigen was performed. The flock-level factors for C. abortus prevalence were tested using multivariate logistic regression model. Fifty-five flocks out of 110 presented at least one seropositive animal with an overall prevalence of 50.0% (95%; CI: 40.3%, 59.7%. Ninety-one out of 975 dairy goats examined were seropositive with titers >32, resulting in a frequency of 9.3%. Lend buck for breeding (odds ratio = 2.35; 95% CI: 1.04-5.33 and history of abortions (odds ratio = 3.06; 95% CI: 1.37-6.80 were associated with increased flock prevalence.

  17. Many globally isolated AD hybrid strains of Cryptococcus neoformans originated in Africa.

    Directory of Open Access Journals (Sweden)

    Anastasia P Litvintseva

    2007-08-01

    Full Text Available Interspecific and intervarietal hybridization may contribute to the biological diversity of fungal populations. Cryptococcus neoformans is a pathogenic yeast and the most common fungal cause of meningitis in patients with AIDS. Most patients are infected with either of the two varieties of C. neoformans, designated as serotype A (C. neoformans var. grubii or serotype D (C. neoformans var. neoformans. In addition, serotype AD strains, which are hybrids of these two varieties, are commonly isolated from clinical and environmental samples. While most isolates of serotype A and serotype D are haploid, AD strains are diploid or aneuploid, and contain two sets of chromosomes and two mating type alleles, MATa and MATalpha, one from each of the serotypes. The global population of serotype A is dominated by isolates with the MATalpha mating type (Aalpha; however, about half of the globally analyzed AD strains possess the extremely rare serotype A MATa allele (Aa. We previously described an unusual population of serotype A in Botswana, in which 25% of the strains contain the rare MATa allele. Here we utilized two methods, phylogenetic analysis of three genes and genotyping by scoring amplified fragment length polymorphisms, and discovered that AD hybrid strains possessing the rare serotype A MATa allele (genotype AaDalpha cluster with isolates of serotype A from Botswana, whereas AD hybrids that possess the MATalpha serotype A allele (AalphaDa and AalphaDalpha cluster with cosmopolitan isolates of serotype A. We also determined that AD hybrid strains are more resistant to UV irradiation than haploid serotype A strains from Botswana. These findings support two hypotheses: (i AaDalpha strains originated in sub-Saharan Africa from a cross between strains of serotypes A and D; and (ii this fusion produced hybrid strains with increased fitness, enabling the Botswanan serotype A MATa genome, which is otherwise geographically restricted, to survive, emigrate

  18. Draft Genome Sequences of Three Novel Low-Abundance Species Strains Isolated from Kefir Grain

    Science.gov (United States)

    Kim, Yongkyu; Blasche, Sonja

    2017-01-01

    ABSTRACT We report here the genome sequences of three novel bacterial species strains—Bacillus kefirresidentii Opo, Rothia kefirresidentii KRP, and Streptococcus kefirresidentii YK—isolated from kefir grains collected in Germany. The draft genomes of these isolates were remarkably dissimilar (average nucleotide identities, 77.80%, 89.01%, and 92.10%, respectively) to those of the previously sequenced strains. PMID:28963201

  19. Complete genome sequence of Bifidobacterium breve CECT 7263, a strain isolated from human milk.

    Science.gov (United States)

    Jiménez, Esther; Villar-Tajadura, M Antonia; Marín, María; Fontecha, Javier; Requena, Teresa; Arroyo, Rebeca; Fernández, Leónides; Rodríguez, Juan M

    2012-07-01

    Bifidobacterium breve is an actinobacterium frequently isolated from colonic microbiota of breastfeeding babies. Here, we report the complete and annotated genome sequence of a B. breve strain isolated from human milk, B. breve CECT 7263. The genome sequence will provide new insights into the biology of this potential probiotic organism and will allow the characterization of genes related to beneficial properties.

  20. Colorful microdiversity of Synechococcus strains (picocyanobacteria) isolated from the Baltic Sea

    NARCIS (Netherlands)

    Haverkamp, T.H.A.; Schouten, D.; Doeleman, M.; Wollenzien, U.; Huisman, J.; Stal, L.J.

    2009-01-01

    Synechococcus is a cosmopolitan genus of picocyanobacteria living in the photic zone of freshwater and marine ecosystems. Here, we describe the isolation of 46 closely related picocyanobacterial strains from the Baltic Sea. The isolates showed considerable variation in their cell size and

  1. Complete genome sequence of the Campylobacter iguaniorum strain RM11343, isolated from an alpaca

    Science.gov (United States)

    Campylobacter iguaniorum is a member of the C. fetus group of campylobacters and is one of two Campylobacter taxa isolated from reptiles. This study describes the whole-genome sequence of the C. iguaniorum strain RM11343, which was isolated from a California alpaca fecal sample....

  2. SELECTION OF INDIGENOUS YEAST STRAINS ISOLATED FROM CV. GEWURZTRAMINER FROM ILOK WINEGROWING REGION

    Directory of Open Access Journals (Sweden)

    Dora DUJMOVIĆ

    2008-11-01

    Full Text Available A total of 36 indigenous strains belonging to the Saccharomyces sensu stricto complex isolated from the variety of Gewurztraminer from Ilok winegrowing region were tested with the purpose of making top-quality wines. Microfermentation was applied in order to examine the synthesis of ethyl alcohol and volatile acidity. Killer factor, synthesis of pectinase, ß-glucosidase and H2S was determined in isolated yeast strains. In the selection results, strains RO 1344, RO 1351, RO 1364 were the best evaluated and they should be examined in midifermentation.

  3. [Molecular typing of 12 Brucella strains isolated in Guizhou province in 2010-2013].

    Science.gov (United States)

    Wang, Yue; Chen, Hong; Liu, Ying; Zhou, Jingzhu; Li, Shijun; Hang, Yan; Tang, Guangpeng; Wang, Dingming; Chen, Guichun

    2015-09-01

    To identify and characterize the Brucella strains from Guizhou province in 2010-2013. A total of 12 strains of Brucella suspicious bacteria were isolated in Guizhou province from 2010 to 2013. Four strains (GZLL3, GZLL4, GZLL11 and SH2) were isolated from goat blood samples and eight strains (SH4, GZZY, GZSQ, GZZA, BR13001, BR13004, BR13005 and BR13006) were isolated from blood samples of patient 12 Brucella suspicious strains were identified and characterized using conventional methods. Brucella genus specific gene BCSP31-based PCR (BCSP31-PCR) was used to identify the genus of Brucella and IS711 insert sequence-based PCR (AMOS-PCR) was applied to identify the species of Brucella strains. Goats and patients originated Brucella strains were comparatively analysed using Pulse-field Gel Electrophoresis (PFGE). Both of conventional methods and PCR identified the 12 Brucella suspicious strains as B. melitensis biotype 3. BCSP31-PCR identification results showed that a specific DNA bands (223 bp) were detected in all the 12 strains and positive control samples with no DNA band in negative samples. AMOS-PCR amplified a 731 bp-DNA bands in all the 12 strains, with 731 bp, 498 bp and 275 bp in M5, S2 and A19 strains, respectively, and no DNA band was detected in the negative control samples. PFGE analysis showed that 12 Brucella isolates from patients and goats showed consistent PFGE patterns with the digestion of restriction enzyme Xba I. The epidemic species/type of Brucella in both human and animal in Guizhou province was B. melitensis biotype 3 and goat was the main animal source of infection of brucellosis in Guizhou province.

  4. Molecular characterization of Helicobacter pylori strains isolated from cynomolgus monkeys (M. fascicularis).

    Science.gov (United States)

    Doi, Sonia Q; Kimbason, Tara; Reindel, James; Dubois, Andre

    2005-06-15

    We recently reported the occurrence of natural infection with H. pylori in a group of cynomolgus monkeys with chronic active gastritis and gastric erosions. The goal of the present study was to characterize and to compare strains isolated from animals originating from two different geographical areas. Gross and microscopic pathology determined at the time of necropsy was similar in all animals. H. pylori were isolated from specimens harvested in five monkeys (four from Vietnam and one from the Philippines) with gastritis. Isolates from monkeys bred in Vietnam had a similar DNA fingerprint pattern, which was distinct from that of isolates from a monkey bred in the Philippines. All strains were of the s1a vacA subtype, but all the 'Vietnamese' strains were cagA+ and all but one were iceA1 whereas the 'Philippino' strains were cagA- and iceA2. The sequences of the 16S rRNA of the Vietnamese and Philippino strains shared 98% homology and both clustered with H. pylori sequences present in the NCBI database. In conclusion, cynomolgus monkeys can be naturally colonized by H. pylori, and the strains isolated from these animals appear to vary according to the geographical origin, thus indicating probable infection prior to importation. Since some of the cynomolgus monkeys developed antral erosions during natural infection, we propose that this animal model may be used to investigate the role of H. pylori in ulcerogenesis.

  5. Isolation of Dickeya dadantii strains from potato disease and biocontrol by their bacteriophages.

    Science.gov (United States)

    Soleimani-Delfan, Abbas; Etemadifar, Zahra; Emtiazi, Giti; Bouzari, Majid

    2015-01-01

    One of the most economically important bacterial pathogens of plants and plant products is Dickeya dadantii. This bacterium causes soft rot disease in tubers and other parts of the potato and other plants of the Solanaceae family. The application of restricted host range bacteriophages as biocontrol agents has recently gained widespread interest. This study purposed to isolate the infectious agent of the potato and evaluate its biocontrol by bacteriophages. Two phytopathogenic strains were isolated from infected potatoes, identified based on biochemical and 16S rRNA gene sequencing, and submitted to GenBank as D. dadantii strain pis3 (accession no. HQ423668) and D. dadantii strain sip4 (accession no. HQ423669). Their bacteriophages were isolated from Caspian Sea water by enriching the water filtrate with D. dadantii strains as hosts using spot or overlay methods. On the basis of morphotypes, the isolated bacteriophages were identified as members of the Myoviridae and Siphoviridae families and could inhibit the growth of antibiotic resistant D. dadantii strains in culture medium. Moreover, in Dickeya infected plants treated with bacteriophage, no disease progression was detected. No significant difference was seen between phage-treated and control plants. Thus, isolated bacteriophages can be suggested for the biocontrol of plant disease caused by Dickeya strains.

  6. Widespread pyocyanin over-production among isolates of a cystic fibrosis epidemic strain

    Science.gov (United States)

    Fothergill, Joanne L; Panagea, Stavroula; Hart, Charles A; Walshaw, Martin J; Pitt, Tyrone L; Winstanley, Craig

    2007-01-01

    Background Some isolates of the Liverpool cystic fibrosis epidemic strain of Pseudomonas aeruginosa exhibit an unusual virulence-related phenotype, characterized by over-production of quorum sensing-regulated exoproducts such as pyocyanin and LasA protease. Our aim was to determine the prevalence of this unusual phenotype amongst isolates of the epidemic strain, and to study other intraclonal phenotypic and genotypic variations. Results The unusual phenotype was detected in at least one epidemic strain isolate from the majority of cystic fibrosis patients tested, and can be retained for up to seven years during chronic infection. Multiple sequential isolates of the epidemic strain taken from six patients over a period of up to nine years exhibited a wide range of phenotypes, including different antimicrobial susceptibilities. Our data suggest that each sputum sample contains a mixture of phenotypes and genotypes within the epidemic strain population, including within colony morphotypes. Many isolates exhibit premature (during early rather than late exponential growth) and over-production of pyocyanin, which has a number of toxic effects directly relevant to cystic fibrosis. Conclusion The widespread occurrence of this unusual phenotype suggests that it may play an important role in the success of the epidemic strain. PMID:17521417

  7. Widespread pyocyanin over-production among isolates of a cystic fibrosis epidemic strain

    Directory of Open Access Journals (Sweden)

    Pitt Tyrone L

    2007-05-01

    Full Text Available Abstract Background Some isolates of the Liverpool cystic fibrosis epidemic strain of Pseudomonas aeruginosa exhibit an unusual virulence-related phenotype, characterized by over-production of quorum sensing-regulated exoproducts such as pyocyanin and LasA protease. Our aim was to determine the prevalence of this unusual phenotype amongst isolates of the epidemic strain, and to study other intraclonal phenotypic and genotypic variations. Results The unusual phenotype was detected in at least one epidemic strain isolate from the majority of cystic fibrosis patients tested, and can be retained for up to seven years during chronic infection. Multiple sequential isolates of the epidemic strain taken from six patients over a period of up to nine years exhibited a wide range of phenotypes, including different antimicrobial susceptibilities. Our data suggest that each sputum sample contains a mixture of phenotypes and genotypes within the epidemic strain population, including within colony morphotypes. Many isolates exhibit premature (during early rather than late exponential growth and over-production of pyocyanin, which has a number of toxic effects directly relevant to cystic fibrosis. Conclusion The widespread occurrence of this unusual phenotype suggests that it may play an important role in the success of the epidemic strain.

  8. Selection of oleuropein-degrading lactic acid bacteria strains isolated from fermenting Moroccan green olives

    Energy Technology Data Exchange (ETDEWEB)

    Ghabbour, N.; Lamzira, Z.; Thonart, P.; Cidalia, P.; Markaouid, M.; Asehraoua, A.

    2011-07-01

    A total of 177 strains of lactic acid bacteria (LAB) were isolated from early-stage Moroccan Picholine green olive fermentation, including Lactobacillus plantarum (44.63%), Lactobacillus pentosus (25.99%), Lactobacillus brevis (9.61%) and Pediococcus pentosaceus (19.77%). All the isolates were screened for their tolerance to olive leaf extract and oleuropein. Most of the isolates (85.3%) were found able to degrade oleuropein, when evaluated by either oleuropein or 5-Bromo-4-chloro-3-indolyl {beta}-D-glucuronide (X-Gluc) as substrates. The biodegradation capacity of the selected strains of each species was confirmed by HPLC analysis. (Author).

  9. Immunoproteomics of Brucella abortus RB51 as candidate antigens in serological diagnosis of brucellosis.

    Science.gov (United States)

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

    2014-08-15

    The current brucellosis serodiagnostic assays are chiefly based on detecting anti-LPS (lipopolysaccharide) antibodies. However, cross-reaction with some gram-negative bacteria can occasionally induce due to similar O-polysaccharide (OPS) structure. Therefore, the aim of the present study was to identify new candidate antigens from Brucella abortus RB51, a mutant strain lacking the LPS portion, which might be valuable in brucellosis diagnosis. To detect potential antigens, immobilized pH gradients (IPG) strips with three ranges (pH 3-5.6, 4-7 and 6-11) were applied. After separating the insoluble proteins of B. abortus RB51 using two-dimensional electrophoresis (2-DE), their immunogenicity was evaluated by western blotting using four types of antisera - B. abortus, Yersinia enterocolitica O:9 and Escherichia coli O157:H7-positive, and B. abortus-negative bovine sera. Among the several immunogenic spots, the spots showing specific reactivity with only the B. abortus-positive antisera, were considered as candidate antigens. Overall, eleven immuno-reactive proteins were identified, as follows: Cu/Zn superoxide dismutase, histidinol dehydrogenase, chaperonin DnaK, chaperonin GroES, beta-ketoadipyl CoA thiolase, two-component response regulator, the cell-division protein FtsZ, aldehyde dehydrogenase, 50s ribosomal protein L10 and invasion protein B. These selected highly immunogenic protein spots might be useful as alternative antigens for brucellosis and helpful in reducing the cross-reactivity. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Bacteriological Typing of C. Diphtheriae Strains Recently Isolated in Teheran

    Directory of Open Access Journals (Sweden)

    H. Esterabady

    1963-01-01

    Full Text Available From a total of 600 nose and throat introduction swabs examined for diphtherie, 200 or 33% were positive. Cultures were carfully classified on the basis of morphological appearance and biochemical characteristics into Gravis, Mitis and Intermedius groups.  A special tellurite serum agar was used for colonial appearance. Neill's broth culture was employed for haemolytic tests. The virulence of each culture was examined in laboratory animals by the agar gel precipitation method of Elek. From 200 cultures tested, 138 or 69% were gravis, 5 or 2.5% were intermedius, and 57 or 28% were mitis. 'I'hre.e strains of gravis type and one strain of mitis type were avirulent

  12. Characterization of Streptomyces strain SLO-105 isolated from Lake ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-16

    Nov 16, 2009 ... Actinomycetes represent a large group of Gram- positive filamentous bacteria that currently comprises over forty genera and hundreds of species among which is the important Streptomyces genus (Watve et al., 2001). Members of this group have been isolated from different soils, plant materials, waters and ...

  13. Characterization of Rhizobium strain isolated from the roots of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-10-20

    Oct 20, 2008 ... biofertilizer. Key words: Fenugreek, rhizobium isolation, biochemical analysis, immobilization, enzyme production. INTRODUCTION. Nutrient enrichment of soils by nitrogen fixing symbiotic bacteria present in legumes has been known for cen- turies. Scientific demonstration of this symbiosis was started in ...

  14. Isolation and molecular identification of a Serratia strain from ...

    African Journals Online (AJOL)

    ONOS

    2010-04-05

    Apr 5, 2010 ... suitable agar dishes and used in further experiments. Biochemical tests were selectively done according to the standards in Bergey's. Manual of Systematic Bacteriology (second ed) and The Prokaryotes: A Handbook on the Biology of Bacteria (third ed). The isolated colonies were differentiated by colonial ...

  15. Production Of Α-Galactosidase From Local Isolated Strain Of ...

    African Journals Online (AJOL)

    α- Galactosidase was detected in the culture media of Bacillus circulans isolated from soil of (plants Island in Aswan). The optimum condition for growth and enzyme production were determined where the optimum pH was 6.5 to 7and incubation temperatures was 35 to 40ºC. Maximum activity of α-galactosidase in the ...

  16. Two novel EHEC/EAEC hybrid strains isolated from human infections.

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    Rita Prager

    Full Text Available The so far highest number of life-threatening hemolytic uremic syndrome was associated with a food-borne outbreak in 2011 in Germany which was caused by an enterohemorrhagic Escherichia coli (EHEC of the rare serotype O104:H4. Most importantly, the outbreak strain harbored genes characteristic of both EHEC and enteroaggregative E. coli (EAEC. Such strains have been described seldom but due to the combination of virulence genes show a high pathogenicity potential. To evaluate the importance of EHEC/EAEC hybrid strains in human disease, we analyzed the EHEC strain collection of the German National Reference Centre for Salmonella and other Bacterial Enteric Pathogens (NRC. After exclusion of O104:H4 EHEC/EAEC strains, out of about 2400 EHEC strains sent to NRC between 2008 and 2012, two strains exhibited both EHEC and EAEC marker genes, specifically were stx2 and aatA positive. Like the 2011 outbreak strain, one of the novel EHEC/EAEC harbored the Shiga toxin gene type stx2a. The strain was isolated from a patient with bloody diarrhea in 2010, was serotyped as O59:H-, belonged to MLST ST1136, and exhibited genes for type IV aggregative adherence fimbriae (AAF. The second strain was isolated from a patient with diarrhea in 2012, harbored stx2b, was typed as Orough:H-, and belonged to MLST ST26. Although the strain conferred the aggregative adherence phenotype, no known AAF genes corresponding to fimbrial types I to V were detected. In summary, EHEC/EAEC hybrid strains are currently rarely isolated from human disease cases in Germany and two novel EHEC/EAEC of rare serovars/MLST sequence types were characterized.

  17. Draft Genome Sequences of Mycobacterium setense Type Strain DSM-45070 and the Nonpathogenic Strain Manresensis, Isolated from the Bank of the Cardener River in Manresa, Catalonia, Spain.

    Science.gov (United States)

    Rech, Gabriel; Vilaplana, Cristina; Velasco, Juan; Pluvinet, Raquel; Santín, Sheila; Prat, Cristina; Julián, Esther; Alcaide, Fernando; Comas, Iñaki; Sumoy, Lauro; Cardona, Pere-Joan

    2015-02-05

    We present here the draft genome sequences of two Mycobacterium setense strains. One of them corresponds to the M. setense type strain DSM-45070, originally isolated from a patient with a posttraumatic chronic skin abscess. The other one corresponds to the nonpathogenic M. setense strain Manresensis, isolated from the Cardener River crossing Manresa, Catalonia, Spain. A comparative genomic analysis shows a smaller genome size and fewer genes in M. setense strain Manresensis relative to those of the type strain, and it shows the genome segments unique to each strain. Copyright © 2015 Rech et al.

  18. Isolation and Characterization of Hydrocarbon-Degrading Yeast Strains from Petroleum Contaminated Industrial Wastewater

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    Boutheina Gargouri

    2015-01-01

    Full Text Available Two yeast strains are enriched and isolated from industrial refinery wastewater. These strains were observed for their ability to utilize several classes of petroleum hydrocarbons substrates, such as n-alkanes and aromatic hydrocarbons as a sole carbon source. Phylogenetic analysis based on the D1/D2 variable domain and the ITS-region sequences indicated that strains HC1 and HC4 were members of the genera Candida and Trichosporon, respectively. The mechanism of hydrocarbon uptaking by yeast, Candida, and Trichosporon has been studied by means of the kinetic analysis of hydrocarbons-degrading yeasts growth and substrate assimilation. Biodegradation capacity and biomass quantity were daily measured during twelve days by gravimetric analysis and gas chromatography coupled with mass spectrometry techniques. Removal of n-alkanes indicated a strong ability of hydrocarbon biodegradation by the isolated yeast strains. These two strains grew on long-chain n-alkane, diesel oil, and crude oil but failed to grow on short-chain n-alkane and aromatic hydrocarbons. Growth measurement attributes of the isolates, using n-hexadecane, diesel oil, and crude oil as substrates, showed that strain HC1 had better degradation for hydrocarbon substrates than strain HC4. In conclusion, these yeast strains can be useful for the bioremediation process and decreasing petroleum pollution in wastewater contaminated with petroleum hydrocarbons.

  19. Isolation and Characterization of Hydrocarbon-Degrading Yeast Strains from Petroleum Contaminated Industrial Wastewater.

    Science.gov (United States)

    Gargouri, Boutheina; Mhiri, Najla; Karray, Fatma; Aloui, Fathi; Sayadi, Sami

    2015-01-01

    Two yeast strains are enriched and isolated from industrial refinery wastewater. These strains were observed for their ability to utilize several classes of petroleum hydrocarbons substrates, such as n-alkanes and aromatic hydrocarbons as a sole carbon source. Phylogenetic analysis based on the D1/D2 variable domain and the ITS-region sequences indicated that strains HC1 and HC4 were members of the genera Candida and Trichosporon, respectively. The mechanism of hydrocarbon uptaking by yeast, Candida, and Trichosporon has been studied by means of the kinetic analysis of hydrocarbons-degrading yeasts growth and substrate assimilation. Biodegradation capacity and biomass quantity were daily measured during twelve days by gravimetric analysis and gas chromatography coupled with mass spectrometry techniques. Removal of n-alkanes indicated a strong ability of hydrocarbon biodegradation by the isolated yeast strains. These two strains grew on long-chain n-alkane, diesel oil, and crude oil but failed to grow on short-chain n-alkane and aromatic hydrocarbons. Growth measurement attributes of the isolates, using n-hexadecane, diesel oil, and crude oil as substrates, showed that strain HC1 had better degradation for hydrocarbon substrates than strain HC4. In conclusion, these yeast strains can be useful for the bioremediation process and decreasing petroleum pollution in wastewater contaminated with petroleum hydrocarbons.

  20. High-frequency rugose exopolysaccharide production by Vibrio cholerae strains isolated in Haiti.

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    Mustafizur Rahman

    Full Text Available In October, 2010, epidemic cholera was reported for the first time in Haiti in over 100 years. Establishment of cholera endemicity in Haiti will be dependent in large part on the continued presence of toxigenic V. cholerae O1 in aquatic reservoirs. The rugose phenotype of V. cholerae, characterized by exopolysaccharide production that confers resistance to environmental stress, is a potential contributor to environmental persistence. Using a microbiologic medium promoting high-frequency conversion of smooth to rugose (S-R phenotype, 80 (46.5% of 172 V. cholerae strains isolated from clinical and environmental sources in Haiti were able to convert to a rugose phenotype. Toxigenic V. cholerae O1 strains isolated at the beginning of the epidemic (2010 were significantly less likely to shift to a rugose phenotype than clinical strains isolated in 2012/2013, or environmental strains. Frequency of rugose conversion was influenced by incubation temperature and time. Appearance of the biofilm produced by a Haitian clinical rugose strain (altered biotype El Tor HC16R differed from that of a typical El Tor rugose strain (N16961R by confocal microscopy. On whole-genome SNP analysis, there was no phylogenetic clustering of strains showing an ability to shift to a rugose phenotype. Our data confirm the ability of Haitian clinical (and environmental strains to shift to a protective rugose phenotype, and suggest that factors such as temperature influence the frequency of transition to this phenotype.

  1. Phenotypic and molecular identification of Fonsecaea pedrosoi strains isolated from chromoblastomycosis patients in Mexico and Venezuela.

    Science.gov (United States)

    Carolina Rojas, O; León-Cachón, Rafael B R; Pérez-Maya, Antonio Alí; Aguirre-Garza, Marcelino; Moreno-Treviño, María G; González, Gloria M

    2015-05-01

    Chromoblastomycosis is a chronic granulomatous disease caused frequently by fungi of the Fonsecaea genus. The objective of this study was the phenotypic and molecular identification of F. pedrosoi strains isolated from chromoblastomycosis patients in Mexico and Venezuela. Ten strains were included in this study. For phenotypic identification, we used macroscopic and microscopic morphologies, carbohydrate assimilation test, urea hydrolysis, cixcloheximide tolerance, proteolitic activity and the thermotolerance test. The antifungal activity of five drugs was evaluated against the isolates. Molecular identification was performed by sequencing the internal transcribed spacer (ITS) ribosomal DNA regions of the isolated strains. The physiological analysis and morphological features were variable and the precise identification was not possible. All isolates were susceptible to itraconazole, terbinafine, voriconazole and posaconazole. Amphotericin B was the least effective drug. The alignment of the 559-nucleotide ITS sequences from our strains compared with sequences of GenBank revealed high homology with F. pedrosoi (EU285266.1). In this study, all patients were from rural areas, six from Mexico and four from Venezuela. Ten isolates were identified by phenotypic and molecular analysis, using ITS sequence and demonstrated that nine isolates from Mexico and Venezuela were 100% homologous and one isolate showed a small genetic distance. © 2015 Blackwell Verlag GmbH.

  2. Phylogeny of bacteria isolated from Rhabditis sp. (Nematoda) and identification of novel entomopathogenic Serratia marcescens strains.

    Science.gov (United States)

    Tambong, James T

    2013-02-01

    Twenty-five bacterial strains isolated from entomopathogenic nematodes were characterized to the genus level by 16S rRNA phylogeny and BLAST analyses. Bacteria strains isolated could be affiliated with seven genera. Microbacterium-like isolates phylogenetically affiliated with M. oxydans while those of Serratia were highly similar to S. marcescens. 16S rRNA sequences of Bacillus isolates matched those of both B. mycoides and B. weihenstephanesis. One isolate each matched Pseudomonas mosselii, Rheinheimera aquimaris, Achromobacter marplatensis, or Staphylococcus hominis. Serratia isolates were examined further for their pathogenicity to Galleria mellonella larvae. All the Serratia isolates exhibited potent pathogenicity toward G. mellonella larvae and possessed a metalloprotease gene encoding for a novel serralysin-like protein. The nucleotide sequence of the metalloprotease gene had 60 synonymous and 8 nonsynonymous substitutions when compared to the closest genBank entry, S. marcescens E-15, with an insertion of a new aspartic acid residue. Tajima's test for equality of evolutionary rate was significant between the metalloprotease gene sequence of S. marcescens strain DOAB 216-82 (this study) and strain E-15. This new insecticidal metalloprotease gene and/or its product could have applications in agricultural biotechnology.

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

    Science.gov (United States)

    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. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Probiotic features of Lactobacillus strains isolated from Ragusano and Pecorino Siciliano cheeses.

    Science.gov (United States)

    Caggia, C; De Angelis, M; Pitino, I; Pino, A; Randazzo, C L

    2015-09-01

    In the present study 177 Lactobacillus spp. strains, isolated from Ragusano and Pecorino Siciliano cheeses, were in vitro screened for probiotic traits, and their characteristics were compared to those of Lactobacillus rhamnosus GG, commercial strain. Based on acidic and bile salt resistance, thirteen Lactobacillus strains were selected. The multiplex-PCR application revealed that nine strains belonged to L. rhamnosus species and four to Lactobacillus paracasei species. All selected strains were further investigated for transit tolerance in simulated upper gastrointestinal tract (GI), for adhesion capacity to human intestinal cell lines, for hydrophobicity, for co-aggregation and auto-aggregation and for antimicrobial activities. Moreover, antibiotic resistance, hemolytic and bile salt hydrolase activities were investigated for safety assessment. Viable counts after simulated gastric and duodenal transit revealed that overall the selected lactobacilli tolerated better pancreatic juice and bile salts than acidic juice. In particular, three L. rhamnosus strains (FS10, FS2, and PS11) and one L. paracasei strain (PM8) increased their cell density after the simulated GI transit. The same strains showed also high percentage of auto-aggregation and co-aggregation with Escherichia coli. All strains were effective against both Staphylococcus aureus and E. coli and variability was achieved versus Listeria monocytogenes and Enterococcus faecalis used as pathogenic indicator strains. Different behavior was revealed by strains for adhesion ability and hydrophobicity, which are not always linked each other and are strongly strain-dependent. From the safety point of view, no isolate showed hemolytic and bile salt hydrolase activities, except one, and most of the strains were sensitive to a broad range of clinical antibiotics. This work showed that the L. rhamnosus FS10 and the L. paracasei PM8 are good promising probiotic candidates for further in vivo investigations

  5. POTENTIAL PRODUCTION OF CYCLOPIAZONIC ACID BY PENICILLIUM CAMEMBERTI STRAINS ISOLATED FROM CAMEMBERT TYPE CHEESE

    Directory of Open Access Journals (Sweden)

    Miroslava Císarová

    2012-10-01

    Full Text Available The aim of this study was to isolate the strains of fungi from Camembert type cheese, identify them and to test isolated strains of Penicillium camemberti for their ability to produce cyclopiazonic acid. The description of micro- and macromorphological features was used for identification of Penicillium camemberti strains. Strains were subsequently in vitro tested on their potential ability to produce mycotoxin cyclopiazonic acid (CPA. All of the 14 strains of Penicillium camemberti, which were obtained from 20 samples of Camembert type cheese, were cultivated 7, 14, 21, 27 and 30 days on CYA medium at 10±1°C, 15±1°C and 25±1°C in the dark. For determination of CPA production ability by P. camemberti isolates in vitro was TLC used. After 7 days of cultivation cyclopiazonic acid was produced only by 5 from 14 strains cultivated at all cultivation temperatures. After 14 and 21 days of cultivation was CPA produced by 6 strains at all of cultivation temperatures. After 27 and 30 days of cultivation was CPA identified in 7 strains cultivated at all temperatures of cultivation. The other strains also produced mycotoxin, however, not at each temperature. The most productive at all temperatures and after all days were 5 out of 14 tested strains (S9, S10, S13, S18 and S19. Strains S6 and S16 did not produce CPA at any temperature. The lowest production after all days of cultivation was found at 10±1 °C (44% and the highest at 25±1 °C (85%.

  6. Draft genome sequences of eight bacteria isolated from the indoor environment: Staphylococcus capitis strain H36, S. capitis strain H65, S. cohnii strain H62, S. hominis strain H69, Microbacterium sp. strain H83, Mycobacterium iranicum strain H39, Plantibacter sp. strain H53, and Pseudomonas oryzihabitans strain H72.

    Science.gov (United States)

    Lymperopoulou, Despoina S; Coil, David A; Schichnes, Denise; Lindow, Steven E; Jospin, Guillaume; Eisen, Jonathan A; Adams, Rachel I

    2017-01-01

    We report here the draft genome sequences of eight bacterial strains of the genera Staphylococcus, Microbacterium, Mycobacterium, Plantibacter, and Pseudomonas. These isolates were obtained from aerosol sampling of bathrooms of five residences in the San Francisco Bay area. Taxonomic classifications as well as the genome sequence and gene annotation of the isolates are described. As part of the "Built Environment Reference Genome" project, these isolates and associated genome data provide valuable resources for studying the microbiology of the built environment.

  7. Isolation and identification of a distinct strain of Culex Flavivirus from mosquitoes collected in Mainland China

    Directory of Open Access Journals (Sweden)

    Huanyu Wang

    2012-03-01

    Full Text Available Abstract Background Culexflavivirus (CxFV is an insect specific virus that has been isolated from Culexpipiens, Culexquinquefasciatus, Culextritaeniorhynchus and other Culex mosquitoes. It is a novel flavivirus isolated in Asia, North America, Central America and Africa. Phylogenetic analysis indicates that, based on the envelope gene (E gene sequence, the worldwide CxFV strains can be divided into two genotypes. Result A virus (SDDM06-11 was isolated from Culexpipiens collected in Shandong Province, China in 2006. The strain caused cytopathic effect (CPE in Aedesalbopictus (C6/36 cells by 3 days post-infection and immunofluorescence assay (IFA showed a reaction with Japanese encephalitis virus (JEV polyclonal antibodies. Phylogenetic analysis of the E gene sequence showed CxFV formed two genotypes with the SDDM06-11 strain assigned to genotype 1. Analysis of the E gene nucleotide homology showed the virus possessed characteristic amino acids at specific sites. The nucleotide homology of the open reading frame (ORF was 94.8%-95.1% between SDDM06-11 and isolates from Japan, Iowa and Texas, and 90.2%-90.5% between SDDM06-11 and isolates from Uganda and Mexico. Conclusion In this paper we report the first isolation and identification of an isolate of CxFV in mainland China. Phylogenetic analysis indicates the isolate belongs to genotype 1. Our findings provide insight into the occurrence of CxFV in Culex mosquito populations and its distribution on a global scale.

  8. Probiotic potential of selected lactic acid bacteria strains isolated from Brazilian kefir grains.

    Science.gov (United States)

    Leite, A M O; Miguel, M A L; Peixoto, R S; Ruas-Madiedo, P; Paschoalin, V M F; Mayo, B; Delgado, S

    2015-06-01

    A total of 34 lactic acid bacteria isolates from 4 different Brazilian kefir grains were identified and characterized among a group of 150 isolates, using the ability to tolerate acidic pH and resistance to bile salts as restrictive criteria for probiotic potential. All isolates were identified by amplified ribosomal DNA restriction analysis and 16S rDNA sequencing of representative amplicons. Eighteen isolates belonged to the species Leuconostoc mesenteroides, 11 to Lactococcus lactis (of which 8 belonged to subspecies cremoris and 3 to subspecies lactis), and 5 to Lactobacillus paracasei. To exclude replicates, a molecular typing analysis was performed by combining repetitive extragenic palindromic-PCR and random amplification of polymorphic DNA techniques. Considering a threshold of 90% similarity, 32 different strains were considered. All strains showed some antagonistic activity against 4 model food pathogens. In addition, 3 Lc. lactis strains and 1 Lb. paracasei produced bacteriocin-like inhibitory substances against at least 2 indicator organisms. Moreover, 1 Lc. lactis and 2 Lb. paracasei presented good total antioxidative activity. None of these strains showed undesirable enzymatic or hemolytic activities, while proving susceptible or intrinsically resistant to a series of clinically relevant antibiotics. The Lb. paracasei strain MRS59 showed a level of adhesion to human Caco-2 epithelial cells comparable with that observed for Lactobacillus rhamnosus GG. Taken together, these properties allow the MRS59 strain to be considered a promising probiotic candidate. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. Isolation from cattle of a prion strain distinct from that causing bovine spongiform encephalopathy.

    Directory of Open Access Journals (Sweden)

    Vincent Béringue

    2006-10-01

    Full Text Available To date, bovine spongiform encephalopathy (BSE and its human counterpart, variant Creutzfeldt-Jakob disease, have been associated with a single prion strain. This strain is characterised by a unique and remarkably stable biochemical profile of abnormal protease-resistant prion protein (PrP(res isolated from brains of affected animals or humans. However, alternate PrP(res signatures in cattle have recently been discovered through large-scale screening. To test whether these also represent separate prion strains, we inoculated French cattle isolates characterised by a PrP(res of higher apparent molecular mass--called H-type--into transgenic mice expressing bovine or ovine PrP. All mice developed neurological symptoms and succumbed to these isolates, showing that these represent a novel strain of infectious prions. Importantly, this agent exhibited strain-specific features clearly distinct from that of BSE agent inoculated to the same mice, which were retained on further passage. Moreover, it also differed from all sheep scrapie isolates passaged so far in ovine PrP-expressing mice. Our findings therefore raise the possibility that either various prion strains may exist in cattle, or that the BSE agent has undergone divergent evolution in some animals.

  10. Identification of novel anti-inflammatory probiotic strains isolated from pulque.

    Science.gov (United States)

    Torres-Maravilla, Edgar; Lenoir, Marion; Mayorga-Reyes, Lino; Allain, Thibault; Sokol, Harry; Langella, Philippe; Sánchez-Pardo, María E; Bermúdez-Humarán, Luis G

    2016-01-01

    Probiotics are live microorganisms which when administered in adequate amounts, confer health benefits on the host. Their use is more and more widespread for both prevention and treatment of diseases, including traveler’s diarrhea and inflammatory bowel diseases (IBDs). In this work, we isolated and characterized novel candidate probiotic strains from pulque (xaxtle), a traditional Mexican alcoholic fermented beverage. A total of 14 strains were obtained from xaxtle samples isolated from three different Mexican regions. Species identification was performed by biochemical methods and 16S rRNA gene targeted PCR. The isolates belonged to the Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus brevis, and Lactobacillus composti phylogenetic groups, with L. brevis being the most dominant group. Bacteria were tested for lysozyme, low pH, and bile acid resistance. Moreover, the strains were tested for adherence to human intestinal epithelial cells and screened for their immunomodulatory properties using a cellular model. Selected bacterial strains with anti-inflammatory properties were then tested in vivo in a dinitro-benzene sulfonic acid (DNBS)-induced chronic colitis mouse model, and weight loss, gut permeability, and cytokine profiles were measured as readouts of inflammation. One of the selected strains, Lactobacillus sanfranciscensis LBH1068, improved mice health as observed by a reduction of weight loss, significant decreases in gut permeability, and cytokine modulation. Altogether, our results highlighted the potential of lactobacilli isolated from pulque and in particular the strain L. sanfranciscensis LBH1068 as a novel probiotic to treat IBD.

  11. Characterization of Yersinia enterocolitica Biotype 1A Strains Isolated from Swine Slaughterhouses and Markets

    Directory of Open Access Journals (Sweden)

    Renata Paixão

    2013-01-01

    Full Text Available Yersinia enterocolitica is an important foodborne pathogen that causes illness in humans and animals. Y. enterocolitica is also the most heterogeneous species of the genus and is divided into distinct serotypes and over six biotypes. Y. enterocolitica biotype 1A strains are classically considered as nonpathogenic; however, some biotype 1A isolates have been considered as causative of gastrointestinal disease, yielding symptoms indistinguishable from those produced by pathogenic biotypes. Even after decades of isolation of clinical strains, the pathogenic mechanisms of these isolates are still not fully understood. In the present study, 122 Yersinia enterocolitica biotype 1A strains isolated from swine slaughterhouses and meat markets in Sao Paulo, Brazil, were characterized according to the presence of the virulence genes ail, virF, and ystA. A total of 94 strains were positive to at least one virulence gene (77.05%, and 67 were positive to all of them (54.92%. Twenty-two strains were submitted to PFGE genotyping resulting in 22 distinct pulsotypes, varying from 50% to 84% of genetic similarity. Any clustering tendency among pulsotypes related to origin, isolation site, or even virulence profile was not observed. The present study reports an important contamination of the environment in swine slaughterhouses, meat markets, and pork, by potentially virulent Y. enterocolitica biotype 1A.

  12. Characterization of Yersinia enterocolitica biotype 1A strains isolated from swine slaughterhouses and markets.

    Science.gov (United States)

    Paixão, Renata; Moreno, Luisa Zanolli; Sena de Gobbi, Débora Dirani; Raimundo, Daniele Cristine; Hofer, Ernesto; Matté, Maria Helena; Ferreira, Thais Sebastiana Porfida; de Moura Gomes, Vasco Tulio; Costa, Barbara Leticia Pereira; Moreno, Andrea Micke

    2013-01-01

    Yersinia enterocolitica is an important foodborne pathogen that causes illness in humans and animals. Y. enterocolitica is also the most heterogeneous species of the genus and is divided into distinct serotypes and over six biotypes. Y. enterocolitica biotype 1A strains are classically considered as nonpathogenic; however, some biotype 1A isolates have been considered as causative of gastrointestinal disease, yielding symptoms indistinguishable from those produced by pathogenic biotypes. Even after decades of isolation of clinical strains, the pathogenic mechanisms of these isolates are still not fully understood. In the present study, 122 Yersinia enterocolitica biotype 1A strains isolated from swine slaughterhouses and meat markets in Sao Paulo, Brazil, were characterized according to the presence of the virulence genes ail, virF, and ystA. A total of 94 strains were positive to at least one virulence gene (77.05%), and 67 were positive to all of them (54.92%). Twenty-two strains were submitted to PFGE genotyping resulting in 22 distinct pulsotypes, varying from 50% to 84% of genetic similarity. Any clustering tendency among pulsotypes related to origin, isolation site, or even virulence profile was not observed. The present study reports an important contamination of the environment in swine slaughterhouses, meat markets, and pork, by potentially virulent Y. enterocolitica biotype 1A.

  13. Antibiotic resistance determinants in a Pseudomonas putida strain isolated from a hospital.

    Directory of Open Access Journals (Sweden)

    Lázaro Molina

    Full Text Available Environmental microbes harbor an enormous pool of antibiotic and biocide resistance genes that can impact the resistance profiles of animal and human pathogens via horizontal gene transfer. Pseudomonas putida strains are ubiquitous in soil and water but have been seldom isolated from humans. We have established a collection of P. putida strains isolated from in-patients in different hospitals in France. One of the isolated strains (HB3267 kills insects and is resistant to the majority of the antibiotics used in laboratories and hospitals, including aminoglycosides, ß-lactams, cationic peptides, chromoprotein enediyne antibiotics, dihydrofolate reductase inhibitors, fluoroquinolones and quinolones, glycopeptide antibiotics, macrolides, polyketides and sulfonamides. Similar to other P. putida clinical isolates the strain was sensitive to amikacin. To shed light on the broad pattern of antibiotic resistance, which is rarely found in clinical isolates of this species, the genome of this strain was sequenced and analysed. The study revealed that the determinants of multiple resistance are both chromosomally-borne as well as located on the pPC9 plasmid. Further analysis indicated that pPC9 has recruited antibiotic and biocide resistance genes from environmental microorganisms as well as from opportunistic and true human pathogens. The pPC9 plasmid is not self-transmissible, but can be mobilized by other bacterial plasmids making it capable of spreading antibiotic resistant determinants to new hosts.

  14. Biochemical and serological characterization of mycoplasma strains isolated from the genital tracts of humans in Nigeria.

    Science.gov (United States)

    Agbakoba, N R; Adetosoye, A I; Adewole, I F

    2006-06-01

    Fifty-five (55) Mycoplasma strains isolated from the genital tracts of humans were biochemically characterized using various biochemical tests and also serologically identified by growth inhibition technique using 5 mycoplasma antisera namely M. hominis PG2 1: M. genitalium G37: M. penetrans GTU54 and 2 strains of M. fermentans PG18 (HRC 6-62-S-170 and MB713-501-069). Biochemically, 43 (78.2%) strains were identified as Mycoplasma hominis, 8 (14.5%) strains as M. fermentans and 4 (7.3%) as M. penetrans. The M. hominis strains hydrolyzed only arginine while the M. fermentans and M. penetrans strains in addition to arginine hydrolysis also broke down glucose fermentatively and oxidatively. The M. fermentans strains showed varying reactions to phosphatase activity and to the reduction of tetrazolium chloride. Serologically, 4 (7.3%) mycoplasma strains were confirmed as M. penetrans GTU54 and of the 8 M. fermentans strains, 4 (7.3%) were identified as M. fermentans PG18 serotype HRC 6-62-S-170 and the other 4 (7.3%) as M. fermentans PG18 serotype MB 713-501-069. Only 13 (30.2%) of the 43 M. hominis strains were identified as M. hominis serotype PG2 1. None was identified as M. genitalium. The heterogeneity of the mycoplasma strains especially M. hominis was observed in this study and the need for the use of multiple antisera in growth inhibition test is hereby supported.

  15. Isolation and determination antimicrobial susceptibility pattern of Enterobacter cloacae strains isolated from consumed powdered infant formula milk in NICU ward

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    Jalal Mardaneh

    2014-11-01

    Full Text Available Background: Enterobacter cloacae is a rod-shaped, gram-negative bacillus, from the family of Enterobacteriaceae. It is an opportunistic pathogen and causes disease in plants and humans (premature and immunocompromised persons of all age groups. The goal of this study was to isolate and determine antimicrobial susceptibility pattern of Enterobacter cloacae strains isolated from consumed powdered infant formula (PIF milk in Neonatal Intensive Care Unit (NICU ward. Material and Methods: In this cross-sectional study, a total of 125 consumed powdered infant formula milk in NICU ward were surveyed. Isolation and Identification of microorganisms was carried out according to FDA method. Antimicrobial susceptibility test was performed using the standard disc diffusion method based on CLSI (2011 recommendations. Results: Enterobacter cloacae was isolated from 2 (1.6% of 125 PIF milk samples. The results showed that isolated strains are sensitive to most antibiotics. All isolates were resistant to amoxicillin. Conclusion: Since the infant formula (PIF samples are unsterile products and contamination could occure during different steps, it is imperative to prepare the infant formula milk foods according to the manufacturer’s instruction and in an aseptic condition. Contamination of PIF only could be reduced or prevented by monitoring the critical control points and taking appropriate action during the processing.

  16. Molecular Detection of Inducible Clindamycin Resistance among Staphylococcal Strains Isolated from Hospital Patients

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    Shadiyeh Abdollahi

    2013-04-01

    Full Text Available Background & Objectives: Macrolide, lincosamide and streptogramin B (MLSB antimicrobial agents are used in the treatment of staphylococcal infections. They prevent the microbial protein synthesis system through binding to 23 S rRNA. The aim of this study was to apply molecular methods to detect inducible clindamycin resistance genes among staphylococcal strains isolated from clinical specimens.   Methods : Two hundred staphylococcus strains were isolated from nose and throat swabs of patients in Toohid and Besat hospitals in Sanandaj . Antimicrobial susceptibilities of isolates were determined using disc diffusion method, agar screen test and D-Test. A multiplex PCR was performed using primers specific for erm (A, B, C, TR genes.   Results: Out of 200 isolates, 18.5 % were MRSA and 32% were MRCNS (methicillin resistant coagulase negative staphylococci. Of 80 erythromycin resistant isolates, 48 were coagulase negative and 32 were S. aureus. Among the 48 coagulase negative staphylococci (CONS isolates, 11.63% expressed the MLSB-inducible phenotypes. Using PCR, the frequency of different genes in the collection of isolates were as follows: ermA 5.41 % , erm B 5.41 % , and erm C 3.13%. The ermTR gene was negative in all isolates. Among the 32 S. aureus isolates, 9.38% expressed the MLSB-nducible phenotype. Using PCR, these isolates harbored erm A (2.22%, ermB (2.22%, ermC (2.22% and ermTR (2.22% .   Conclusion: This is the first study to show the rate of inducible clindamycin clinical isolates of staphylococci harboring erm genes in Sananadaj. It also demonstrated the frequency of erm genes was higher among CONS isolates than S. aureus. This data suggested the transfer of resistance gene from nonpathogenic to pathogenic strains is likely to happen. Therefore, screening and control of these resistance genes is recommended at clinical laboratories.

  17. [Drug susceptibility and homologous analysis on Corynebacterium striatum strains isolated from inpatients].

    Science.gov (United States)

    Wang, Junrui; Du, Xiaoli; Cui, Jinghua; Guo, Sufang; Fu, Quan; Wang, Yanyan; Fan, Wenbing; Han, Yanqiu

    2014-08-26

    To explore the molecular epidemiological features and in vitro susceptibility profile of Corynebacterium striatum strains isolated from different sites of inpatients, and further provide new data and idea for clinicians to better get knowledge of the clinical significance of Corynebacterium striatum. Fourty-five strains of Corynebacterium striatum isolated from different sites of inpatients from November, 2013 to March, 2014 in Affiliated hospital of Inner Mongolian medical university, and microdilution method was employed to do in vitro antibiotics susceptibility test. Saline-cotton swab method was used to sample the surrounding environmental surfaces for Corynebacterium striatum carrying patients, and suspected colonies were further identified and in vitro drug susceptibility test were performed. Pulsed Field Gel Electrophoresis(PFGE) method was used to do molecular typing for 41 isolates of Corynebacterium striatum. Fourty-five isolates of Corynebacterium striatum strains were mainly isolated from neurosurgical unit(21 isolates), respiratory unit(8 isolates) and intensive care unit(8 isolates), 39 isolates of which were isolated from lower respiratory tract. The 45 isolates presented an aggregate distribution in the following 3 months, which were December 2013, January 2014 and February 2014.In vitro antibiotics susceptibility test showed that MIC90 for penecillin, erythromycin, tetracycline, ciprofloxacin and clindamycin were 64 µg/ml or higher and the MIC90 for gentamicin, vancomycin and rifampicin were all 0.5 µg/ml. Only one isolate was sensitive to all of the antibiotics tested, except clindamycin. PFGE typing results showed that 41 isolates were divided into 7 genotypes, among which 0002 type and 0006 type were the predominant types and accounted for 63% (26/41) and 22% (9/41), respectively. The isolates from different patients showed high homology, which were isolated from the same unit during the same periods.For surrounding surfaces sampling

  18. Characterization of Clostridium perfringens Strains Isolated from Healthy and Necrotic Enteritis-Afflicted Broiler Chickens.

    Science.gov (United States)

    Li, Charles; Lillehoj, Hyun S; Gadde, Ujvala Deepthi; Ritter, Don; Oh, SungTaek

    2017-06-01

    Necrotic enteritis (NE) is an important enteric disease in poultry, and Clostridium perfringens (CP) type A strains are the primary etiology. NE is responsible for annual losses of US $6 billion to the poultry industry in the United States. An increase in the incidence of NE has been also associated with withdrawal of antibiotic growth promoters from poultry feed. In this study, CP strains isolated from healthy and NE-afflicted birds were characterized microbiologically and molecularly, and their virulence was experimentally tested in chickens. All strains were hemolytic, lecithinase positive, and identified as CP by biochemical tests. Three distinct colony morphologies were seen in brain-heart infusion media with 0.3% agarose, FeSO 4 , and ZnCl 2 . The CP strains responded differently to iron chelation with 2,2'-bidypinol. PCR toxinotyping showed that all tested strains were alpha toxin-positive, seven (N11, N10, CP1, CP5, CP13, JGS, and Del1) were beta2-toxin-positive, and only one (Del1) was necrotic enteritis toxin B-like-positive. In vivo studies indicated that most isolates, including strain N11 isolated from the normal chicken gut, were sufficiently virulent to produce NE disease in the Eimeria/CP dual infection model. The Del1 and N11 strains merit further investigation to identify their virulence factors and immune-protective antigens.

  19. UPEC strain characterization isolated from Mexican patients with recurrent urinary infections.

    Science.gov (United States)

    Morales-Espinosa, Rosario; Hernandez-Castro, Rigoberto; Delgado, Gabriela; Mendez, Jose Luis; Navarro, Armando; Manjarrez, Angel; Cravioto, Alejandro

    2016-04-28

    Any microorganism is capable of causing urinary tract infections (UTI). However, Uropathogenic Escherichia coli (UPEC) is responsible for the majority of UTI. A variety of virulence genes have been identified in UPEC. Diverse epidemiological studies support that specific subsets of genes are characteristic of each E. coli uropathogenic subtype involved in the development of cystitis, pyelonephritis, and urosepsis. Twenty-three E. coli strains isolated from women with cystitis, 25 from men with prostatitis were characterized according to serotype, virulence genes, PFGE profile and susceptibility antimicrobials. E. coli O25:H4-ST131 was more frequently isolated from cystitis than prostatitis. The majority of cystitis strains presented almost all the virulence genes, contrary to 60% of prostatitis strains. The strains characterized by serology, PFGE and MLST and virulence profile showed that cystitis strains shared a lineage with their own genomic fingerprint, indicating that these strains derived from a common ancestor. The prostatitis strains showed serotypes diverse and much higher degree of genetic diversity, indicating that they are an unrelated group.  More than 50% of isolates were resistant to at least 7 of the antimicrobials tested. Antibiotic-tolerant cells were observed, presenting with more frequency in response chloramphenicol, tetracycline, trimethoprim-sulfometoxazole and dicloxicillin. Persister cells can be detected from the beginning of the infection. The importance of these persister cells that exhibit multidrug tolerance is that a single surviving bacterium can initiate again an infection making it clinical relevant in the chronic urinary tract infection.

  20. Genotypes of Leptospira spp. strains isolated from dogs in Buenos Aires, Argentina.

    Science.gov (United States)

    Grune Loffler, Sylvia; Passaro, Diego; Samartino, Luis; Soncini, Analía; Romero, Graciela; Brihuega, Bibiana

    2014-01-01

    Leptospirosis is an infectious disease of wide global distribution, which is endemic in Argentina. The objective of this study was to obtain the genetic profiles of Leptospira spp. strains isolated from clinical cases of dogs in the province of Buenos Aires by the multiple-locus variable-number tandem repeat analysis (MLVA). Eight isolated canine strains were genotyped by MLVA, obtaining the identical profile of Leptospira interrogans serovar Canicola Hond Utrecht IV in the strains named Dogy and Mayo. The strains named Bel, Sarmiento, La Plata 4581 and La Plata 5478 were identical to the profile of the genotype of L. interrogans serovar Portlandvere MY 1039.The strain named Avellaneda was identical to the genotype profile of L. interrogans serovar Icterohaemorrhagiae RGA and the strain named SB had the same profile as the L. interrogans serovar Pomona Baires genotype and was similar to the profile of serovar Pomona Pomona genotype. It would be useful to include a larger number of isolates from different dog populations in various provinces of Argentina and to characterize the genetic profiles of the strains circulating in the country. The information obtained will be useful for the control of leptospirosis in the dog population. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España. All rights reserved.

  1. Antibiotic resistance of Enterobacteriaceae strains isolated from different animals gastrointestinal tracts

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    Lukáš Hleba

    2015-05-01

    Full Text Available In this study we monitored antibiotic resistance in Enterobacteriaceae strains isolated from different animals gastrointestinal tracts  (GIT. We isolated Enterobacteriaceae from chicken, ducks, lambs, pigs, sheeps, cows and rabbits collected from slovakian farms. Enterobacteriaceae strains were cultivated on MacConkey agar at 35° ± 2°C at 24 hours. Pure cultures of Enterobacteriaceae strains were obtained by four-way streak method on Chromogenic coliform agar. Identification of purified Enterobacteriaceae strains were done by Enterotest 24 and MALDI TOF MS. For susceptibility testing disk diffusion method was used according by EUCAST. We determined the most resistance in Enterobacteriaceae strains against streptomycin, tetracycline, ampicillin, piperecillin, levofloxacine, chloramphenicol and smaller level of resistance against amikacin, ceftriaxone and ofloxacine. Equally we detected resistance to more antibiotics in one strain. The most resistance was Salmonella enterica ser. Typhimurium. Also E. coli was resistance against four antibiotics and Raoultella ornithinolytica too. Antibiotic resistance was found in other isolated strains too.

  2. Antibiotic susceptibility, antibacterial activity and characterisation of Enterococcus faecium strains isolated from breast milk

    Science.gov (United States)

    Kıvanç, Sertaç Argun; Kıvanç, Merih; Yiğit, Tülay

    2016-01-01

    Enterococci, which have useful biotechnological applications, produce bacteriocins, including those that exert anti-Listerial activity. The present study aimed to determine the antibiotic susceptibility patterns and antimicrobial activity of Enterococcus faecium strains isolated from human breast milk. The strains were identified using carbohydrate fermentation tests and ribotyping. Subsequently, the antibacterial activity of the isolates was investigated, and the quantities of lactic acid and hydrogen peroxide produced, and the proteolytic activity of E. faecium, were determined. In addition, biofilm formation by E. faecium strains was assessed. E. faecium strains exhibited antimicrobial activity against food-borne and clinical bacterial isolates. Furthermore, following 24 h incubation, the tested strains exhibited resistance to a pH range of 2.0–9.5 and tolerance of bile acid, lysozyme activity and phenol. Supernatants of the E. faecium TM13, TM15, TM17 and TM18 strains were shown to be effective against Listeria monocytogenes, and were also resistant to heat. Further studies are required in order to determine whether certain strains of E. faecium may be used for the development of novel antibacterial agents. PMID:27602088

  3. Antibacterial activity of Achillea tenuifolia Lam. extract against standard bacteria and isolated strains

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    Sahar Omidpanah

    2016-12-01

    Full Text Available Researchers have been trying to develop new broad-spectrum antibiotics against the infectious diseases caused by bacteria, fungi, viruses, and parasites for many decades. Prolonged usage of the antibiotics has led to the emergence of drug resistance among bacteria; therefore, there is a tremendous need for novel antimicrobial agents from different sources such as plants which are used in traditional medicine. The aim of this study was to evaluate antibacterial effect of Achillea tenuifolia. The plant material was extracted by maceration method using methanol three times at room temperature. The extract was concentrated after removing the solvent by rotary evaporator and then lyophilized using freeze dryer. Inhibitory effect of the extract was examined against four standard bacteria strains and two isolated strains from diseased hen using disk diffusion method and microdilution method to evaluate their inhibition zone diameter (IZD and minimum inhibitory concentration (MIC, respectively. The results showed that the extract of the plant was active against standard strains including Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis with IZDs of 10.3±0.5, 14±0.0, 12±0.0 and 11.6±0.5, respectively. However, growths of isolated strains were not inhibited in the presence of the extract. Although, the growths of isolated strains were not inhibited by the plant extract, the standard strains were moderately susceptible to the extract; among those P. aeroginosa was more sensible than other tested strains

  4. THE STUDY OF ANTIBIOTIC- AND FAGOSENSITIVITY OF NOSOCOMIAL STRAINS BACTERIA ISOLATED FROM TRANSPLANTED PATIENTS

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    N. I. Gabrielan

    2011-01-01

    Full Text Available Antibiotic and fagosensitivity most etiologically important nosocomial strains of bacteria – Pseudomonas aeru- ginosa, Klebsiella pneumoniae, E. coli, Proteus spp., Staphylococcus spp. were studied. Multiple drug-resistant bacteria as gram-positive and gram-negative, isolated from 8 substrates, had been demonstrated. With regard to the sensitivity of Pseudomonas aeruginosa >40% was observed in 40–50% of the strains to aminoglycosides – aztreonam, amikacin, netilmicin, and only 23–25% of the strains – to gentamicin and levofloxacin (an average of antibiotic susceptibility was 27%. All strains of ESBL Klebsiella drew up and were sensitive only to imipenem, meropenem and aminoglycosides. Specific phages lysed 43–48% of the strains Pseudomonas aeruginosa and Klebsiella pneumoniae, E. coli, Pro- teus spp., multidrug resistant strains of Staphylococcus spp. It is proposed to introduce the use of phages in clinical practice. 

  5. Isolates of Citrus tristeza virus that overcome Poncirus trifoliata resistance comprise a novel strain.

    Science.gov (United States)

    Harper, S J; Dawson, T E; Pearson, M N

    2010-04-01

    The economically important rootstock species Poncirus trifoliata is resistant to most isolates of Citrus tristeza virus (CTV), but not to members of the CTV resistance-breaking (RB) strain presently found in New Zealand. In this study, five known and suspected RB isolates were separated from field mixtures, and their genomes were sequenced in full. It was found that the RB isolates are members of a single phylogenetically distinct clade with an average of 90.3% genomic nucleotide sequence identity to the closest extant isolate, T36. These isolates also show evidence of multiple recombination events throughout their evolutionary history, with T36, T30 and VT-like isolates, and with each other. Finally, the genomic sequences of these isolates show that several genes contain unique polymorphisms that may or may not be involved in overcoming resistance. These data will aid in the understanding of host-virus interactions, and the mechanism of resistance in P. trifoliata.

  6. Epidemiological typing of Acanthamoeba strains isolated from keratitis cases in Belgium.

    Science.gov (United States)

    De Jonckheere, J F

    2003-01-01

    From the corneas of nine keratitis patients and from their contact lenses, contact lens boxes and saline solutions, 15 strains of Acanthamoeba have been isolated. An Acanthamoeba strain was isolated from the swimming pool where one of the patients swam, while in the tapwater of the houses of three patients investigated, no Acanthamoeba could be detected. All the Acanthamoeba isolates from the cornea belong to genotype T4, but are different subtypes of T4. The Acanthamoeba detected on the contact lenses (and/or associated paraphernalia) of a patient are of the same subtype as that isolated from the cornea. The only Acanthamoeba strain isolated from a contact lens which was not related to an Acanthamoeba keratitis infection proved to be another genotype. A strain of Hartmannella from a cornea and two vahlkampfiids isolated from contact lenses had no connection with keratitis. This study confirms that, as found elsewhere, only Acanthamoeba genotype T4 of the 12 known Acanthamoeba genotypes is responsible for keratitis in Belgium. Most cases of Acanthamoeba keratitis cases are due to poor hygiene in the treatment (cleaning and storage) of contact lenses.

  7. QUANTIFICATION OF SIDEROPHORE AND HEMOLYSIN FROM STACHYBOTRYS CHARTARUM STRAINS, INCLUDING A STRAIN ISOLATED FROM THE LUNG OF A CHILD WITH PULMONARY HEMORRHAGE AND HEMOSIDEROSIS

    Science.gov (United States)

    A strain of Stachybotrys chartarum was recently isolated from the lung of a pulmonary hemorrhage and hemosiderosis (PH) patient in Texas (designated the Houston strain). This is the first time that S. chartarum has been isolated from the lung of a PH patient. In this study, the ...

  8. Whole-Genome Sequences of 14 Strains of Bradyrhizobium canariense and 1 Strain of Bradyrhizobium japonicum Isolated from Lupinus spp. in Algeria.

    Science.gov (United States)

    Chekireb, Djamel; Crovadore, Julien; Brachmann, Andreas; Chablais, Romain; Cochard, Bastien; Lefort, François

    2017-07-20

    We report here the whole-genome sequences of 14 strains of Bradyrhizobium canariense, isolated from root nodules of Lupinus microanthus and Lupinus angustifolius, and 1 strain of Bradyrhizobium japonicum isolated from root nodules from Lupinus angustifolius in Algeria. These sequences add to the known diversity of this agronomically important genus. Copyright © 2017 Chekireb et al.

  9. The cultural and biochemical characters of Streptococcus milleri strains isolated from human sources

    Science.gov (United States)

    Ball, Lyn C.; Parker, M. T.

    1979-01-01

    isolations from the bloodstream of patients suffering from systemic infections were β haemolytic and only 18% possessed a group antigen, but a considerably greater proportion of those from visceral abscesses were β haemolytic (28%). Among isolations from superficial lesions in some body sites there were considerably greater proportions of β-haemolytic and groupable strains; thus, nearly one-half of those isolated from the abdomen other than the female genital tract were β-haemolytic and over one-half were groupable. On the other hand, strains from the teeth and gums were nearly always non-haemolytic and ungroupable, and most vaginal isolations were of non-haemolytic strains with a wide sugar-fermentation pattern. PMID:762404

  10. Isolation and characterization of novel bacterial strains exhibiting ligninolytic potential

    Directory of Open Access Journals (Sweden)

    Bandounas Luaine

    2011-10-01

    Full Text Available Abstract Background To expand on the range of products which can be obtained from lignocellulosic biomass, the lignin component should be utilized as feedstock for value-added chemicals such as substituted aromatics, instead of being incinerated for heat and energy. Enzymes could provide an effective means for lignin depolymerization into products of interest. In this study, soil bacteria were isolated by enrichment on Kraft lignin and evaluated for their ligninolytic potential as a source of novel enzymes for waste lignin valorization. Results Based on 16S rRNA gene sequencing and phenotypic characterization, the organisms were identified as Pandoraea norimbergensis LD001, Pseudomonas sp LD002 and Bacillus sp LD003. The ligninolytic capability of each of these isolates was assessed by growth on high-molecular weight and low-molecular weight lignin fractions, utilization of lignin-associated aromatic monomers and degradation of ligninolytic indicator dyes. Pandoraea norimbergensis LD001 and Pseudomonas sp. LD002 exhibited best growth on lignin fractions, but limited dye-decolourizing capacity. Bacillus sp. LD003, however, showed least efficient growth on lignin fractions but extensive dye-decolourizing capacity, with a particular preference for the recalcitrant phenothiazine dye class (Azure B, Methylene Blue and Toluidene Blue O. Conclusions Bacillus sp. LD003 was selected as a promising source of novel types of ligninolytic enzymes. Our observations suggested that lignin mineralization and depolymerization are separate events which place additional challenges on the screening of ligninolytic microorganisms for specific ligninolytic enzymes.

  11. Bioprospecting of Leuconostoc mesenteroides strains isolated from Algerian raw camel and goat milk for technological properties useful as adjunct starters

    OpenAIRE

    ZAROUR, K.; BENMECHERNENE, Z.; HADADJI, M.; MOUSSA-BOUDJEMAA, B.; HENNI, D.; Kihal, M.

    2012-01-01

    Leuconostoc species are lactic acid bacteria widely used in milk fermentation. Based on morphological, physiological and biochemical analysis, 18 strains of Leuconostoc mesenteroides were isolated and identified from 10 samples of goat's milk and camel's milk. Strains were identified as follows 09 strains of L. mesenteroides subsp. mesenteroides and 09 strains of L. mesenteroides subsp. dextranicum The results of technological tests of the strains showed that strains produced dextran, car...

  12. Characteristics of multidrug-resistant Acinetobacter baumannii strains isolated in Geneva during colonization or infection.

    Science.gov (United States)

    Cherkaoui, Abdessalam; Emonet, Stéphane; Renzi, Gesuele; Schrenzel, Jacques

    2015-09-11

    This study determined the antibiotic susceptibility profile and genetic mechanisms of β-lactam resistance in 27 clinical strains of Acinetobacter baumannii isolated at the University Hospitals of Geneva, Switzerland. The antimicrobial susceptibility testing was performed using Etest and the disc diffusion method in accordance with CLSI guidelines. All of the strains were defined as multi-drug resistant (MDR) and were susceptible to colistin and moderately susceptible to tigecycline. Uniplex PCR assays were used to detect the following β-lactamase genes: four class D carbapenem-hydrolysing oxacillinases (blaOXA-51, blaOXA-23, blaOXA-24 and blaOXA-58), four class B metallo-β-lactamases genes (blaIMP, blaVIM, blaSPM and blaNDM) and two class A carbapenemases (blaKPC and blaGES). All of the strains were positive for blaOXA-51 (intrinsic resistance), 14/27 strains carried blaOXA-23, 2/27 strains carried a blaOXA-24-like gene, and 4/27 strains had a blaOXA-58 gene. blaGES-11 was found in three strains, and NDM-1-harbouring strains were identified in three patients. All of the A. baumannii isolates were typed by rep-PCR (DiversiLab) and excluded any clonality. Altogether, this analysis suggests a very high genetic diversity of imported MDR A. baumannii.

  13. Probiotic properties of Lactobacillus and Bifidobacterium strains isolated from porcine gastrointestinal tract.

    Science.gov (United States)

    Kim, Pyoung Il; Jung, Min Young; Chang, Young-Hyo; Kim, Saehun; Kim, Seong-Jae; Park, Yong-Ha

    2007-04-01

    One strain of Lactobacillus salivarius, two strains of Lactobacillus reuteri and Lactobacillus amylovorus, and two strains of Bifidobacterium thermacidophilum with antagonistic effect against Clostridium perfringens were isolated from porcine gastrointestinal tract. Isolates were assayed for their ability to survive in synthetic gastric juice at pH 2.5 and were examined for their ability to grow on agar plate containing porcine bile extract. There was a large variation in the survival of the isolates in gastric juice and growth in the medium containing 0.3% (w/v) bile. L. salivarius G11 and L. amylovorus S6 adhered to the HT-29 epithelial cell line. Cell-free supernatant of L. amylovorus S6 showed higher antagonistic activity as effective as the antibiotics such as neomycin, chlortetracycline, and oxytetracycline against bacterial pathogens including C. perfringens, Salmonella typhimurium, Staphylococcus aureus, Vibrio cholerae, Edwardsiella tarda, and Aeromonas salmonicida subsp. salmonicida.

  14. Phenotypic and Genotypic Antimicrobial Resistance of Lactococcus Sp. Strains Isolated from Rainbow Trout (Oncorhynchus Mykiss

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    Ture Mustafa

    2015-04-01

    Full Text Available A current profile of antimicrobial resistance and plasmid of 29 Lactococcus garvieae and one Lactococcus lactis strains isolated from rainbow trouts (Oncorhynchus mykiss from farms throughout Turkey were investigated. All isolates were sensitive to penicillin G (90%, ampicillin (86.7%, florfenicol (83.3%, amoxicillin (80.1%, and tetracycline (73.4%, and resistant to trimethoprim+sulfamethoxazole (86.6% and gentamycin (46.6% by disc diffusion method. Twenty-eight (93% isolates had two to seven antibiotic resistance genes (ARGs determined by PCR. The most prevalent ARGs were tetracycline (tetB, erythromycin (ereB, and β-lactam (blaTEM. Bacterial strains were also screened for plasmid DNA by agarose gel electrophoresis and two strains harboured plasmids, with sizes ranging from 3 to 9 kb.

  15. Longitudinal genotyping of Candida dubliniensis isolates reveals strain maintenance, microevolution, and the emergence of itraconazole resistance.

    LENUS (Irish Health Repository)

    Fleischhacker, M

    2010-05-01

    We investigated the population structure of 208 Candida dubliniensis isolates obtained from 29 patients (25 human immunodeficiency virus [HIV] positive and 4 HIV negative) as part of a longitudinal study. The isolates were identified as C. dubliniensis by arbitrarily primed PCR (AP-PCR) and then genotyped using the Cd25 probe specific for C. dubliniensis. The majority of the isolates (55 of 58) were unique to individual patients, and more than one genotype was recovered from 15 of 29 patients. A total of 21 HIV-positive patients were sampled on more than one occasion (2 to 36 times). Sequential isolates recovered from these patients were all closely related, as demonstrated by hybridization with Cd25 and genotyping by PCR. Six patients were colonized by the same genotype of C. dubliniensis on repeated sampling, while strains exhibiting altered genotypes were recovered from 15 of 21 patients. The majority of these isolates demonstrated minor genetic alterations, i.e., microevolution, while one patient acquired an unrelated strain. The C. dubliniensis strains could not be separated into genetically distinct groups based on patient viral load, CD4 cell count, or oropharyngeal candidosis. However, C. dubliniensis isolates obtained from HIV-positive patients were more closely related than those recovered from HIV-negative patients. Approximately 8% (16 of 194) of isolates exhibited itraconazole resistance. Cross-resistance to fluconazole was only observed in one of these patients. Two patients harboring itraconazole-resistant isolates had not received any previous azole therapy. In conclusion, longitudinal genotyping of C. dubliniensis isolates from HIV-infected patients reveals that isolates from the same patient are generally closely related and may undergo microevolution. In addition, isolates may acquire itraconazole resistance, even in the absence of prior azole therapy.

  16. /sup 125/I-peptide mapping of protein III isolated from four strains of Neisseria gonorrhoeae

    Energy Technology Data Exchange (ETDEWEB)

    Judd, R.C.

    1982-08-01

    Gonococcal outer-membrane protein I (PI) and PIII were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from reduced and unreduced whole-cell and outer-membrane lysates of four strains of nonpiliated (P-), transparent (O-) Neisseria gonorrhoeae. These proteins were radioiodinated and digested with alpha-chymotrypsin. The resultant /sup 125/I-peptides were then resolved by high-voltage thin-layer electrophoresis, followed by ascending thin-layer chromatography, and visualized by autoradiography. Results corroborated previous observations regarding the structural relationships of PIs having different apparent subunit molecular weights. All PIIIs had very similar apparent primary structures, regardless of the strain from which they were isolated, the source (i.e., whole cells or outer membranes), or the reduction state of the sodium dodecyl sulfate lysates. By the techniques used, it appeared that PIII is structurally similar in all of the gonococcal strains studied, even though each strain had structurally unique PIs.

  17. [The susceptibility of Proteus mirabilis strains isolated from white stork (Ciconia ciconia)].

    Science.gov (United States)

    Kwiecińska-Piróg, Joanna; Bogiel, Tomasz; Gospodarek, Eugenia; Kasprzak, Mariusz; Jerzak, Leszek; Kamiński, Piotr

    2011-01-01

    Proteus sp. rods are ubiquitous bacteria, widespread in the environment and classified also as opportunistic human pathogens. The aim of our study was to evaluate susceptibility of Proteus mirabilis strains isolated from white stork (Ciconia ciconia) regarding as his natural bacterial flora, compare and discuss their results with data obtained from scientific literature for clinical strains of the same species. Susceptibility of 59 P. mirabilis strains was estimated for 27 antimicrobials using disc-diffusion method and the ability to produce extended spectrum beta-lactamases was evaluated by double disc synergy test. Environmental P. mirabilis strains isolated from white stork were assessed as more susceptible to most of the examined antimicrobials and production of extended spectrum beta-lactamases was not noted amongst them.

  18. Biochemical and serological study of two mycoplasma strains isolated from geese.

    Science.gov (United States)

    Varga, Z; Stipkovits, L; Dobos-Kovács, M; Czifra, G

    1989-01-01

    2 mycoplasma strains were isolated, one from the phallic lymph of a gander and the other from a cloacal swab of a laying goose. The strains proved to be different from mycoplasma species isolated from geese before. Strain No. 1223 is a glucose-negative and arginine-negative species belonging to the genus Mycoplasma. In the growth inhibition test, it fails to react with hyperimmune sera raised in rabbits against the presently known mycoplasma species of avian origin nor with sera produced against mammalian mycoplasma species sharing its biochemical properties. Strain No. 1225 belongs to the digitonin resistant Acholeplasmataceae family. It is glucose-positive and aesculin-positive. It is negative by all the other tests and fails to react with sera produced against the presently known acholeplasma species.

  19. Effect of CuO Nanoparticles over Isolated Bacterial Strains from Agricultural Soil

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    Sandra I. Concha-Guerrero

    2014-01-01

    Full Text Available The increased use of the nanoparticles (NPs on several processes is notorious. In contrast the ecotoxicological effects of NPs have been scarcely studied. The main current researches are related to the oxide metallic NPs. In the present work, fifty-six bacterial strains were isolated from soil, comprising 17 different OTUs distributed into 3 classes: Bacilli (36 strains, Flavobacteria (2 strains, and Gammaproteobacteria (18 strains. Copper oxide nanoparticles (CuONPs were synthesized using a process of chemical precipitation. The obtained CuONPs have a spherical shape and primary size less than 17 nm. Twenty-one strains were used to evaluate the cytotoxicity of CuONPs and 11 of these strains showed high sensibility. Among those 11 strains, 4 (Brevibacillus laterosporus strain CSS8, Chryseobacterium indoltheticum strain CSA28, and Pantoea ananatis strains CSA34 and CSA35 were selected to determine the kind of damage produced. The CuONPs toxic effect was observed at expositions over 25 mg·L−1 and the damage to cell membrane above 160 mg·L−1. The electron microscopy showed the formation of cavities, holes, membrane degradation, blebs, cellular collapse, and lysis. These toxic effects may probably be due to the ions interaction, the oxide-reduction reactions, and the generation of reactive species.

  20. IncH plasmids in Escherichia coli strains isolated from broiler chicken carcasses.

    OpenAIRE

    Chaslus-Dancla, E; Lafont, J P

    1985-01-01

    Plasmids conferring tellurite resistance were transferred at low temperature (27 degrees C) from Escherichia coli strains isolated from chicken carcasses at the time of slaughter and after storage. They belonged to group IncH, as evidenced by their large molecular weight and incompatibility with plasmid pIP233. E. coli strains contaminating chickens meat can thus represent a source of IncH plasmids in the food chain of humans.

  1. Zearalenone and Its Derivatives ?-Zearalenol and ?-Zearalenol Decontamination by Saccharomyces cerevisiae Strains Isolated from Bovine Forage

    OpenAIRE

    Keller, Luiz; Abrunhosa, Lu?s; Keller, Kelly; Rosa, Carlos Alberto; Cavaglieri, Lilia; Ven?ncio, Armando

    2015-01-01

    Zearalenone (ZEA) and its derivatives are mycotoxins with estrogenic effects on mammals. The biotransformation for ZEA in animals involves the formation of two major metabolites, - and -zearalenol (-ZOL and -ZOL), which are subsequently conjugated with glucuronic acid. The capability of Saccharomyces cerevisiae strains isolated from silage to eliminate ZEA and its derivatives -ZOL and -ZOL was investigated as, also, the mechanisms involved. Strains were grown on Yeast Extract-Peptone-Dextrose...

  2. Prevalence of extended spectrum beta lactamases among strains of Pseudomonas aeruginosa isolated from burn patients

    Directory of Open Access Journals (Sweden)

    Mirsalehian

    2008-08-01

    Full Text Available Background: The resistance of Pseudomonas aeruginosa strains to broad spectrum cephalosporins may be mediated by extended spectrum b-lactamases (ESBLs. These enzymes are encoded by different genes located either on chromosome or plasmids. In this study, we determined the antimicrobial resistance patterns of P. aeruginosa isolates and screened for ESBL production. Methods: After isolation from burn patients in Tehran Hospital, identification of P. aeruginosa isolates were assessed using biochemical tests. We then performed disk agar diffusion (DAD according to CLSI guidelines to determine the pattern of antimicrobial resistance. The frequency of ESBLs and prevalence of the OXA-10 and PER-1 genes were determined with combined disk and polymerase chain reaction (PCR methods, respectively. Results: One hundred strains of P. aeruginosa were isolated. The resistance of these strains to cephpodoxime, aztreonam, ciprofloxacin, ofloxacin, ceftazidime, cefepime, imipenem, meropenem, cefotaxime, levofloxacin, piperacilin- tazobactam and ceftriaxon was 100%, 90%, 83%, 92%, 85%, 88%, 63%, 66%, 98%, 89%, 70% and 91%, respectively. Of these, 40 strains (40% were ESBL positive, 29 strains (29% were OXA-10 positive and 18 strains (18% were PER-1 positive. Conclusion: Our results confirm the need for proper antimicrobial therapy in burn hospitals, considering the resistance pattern and frequency of strains producing ESBLs and the presence of the OXA-10 and PER-1 genes. Since an increase in the prevalence of ESBL in P. aeruginosa strains might lead to the transfer of these ESBL genes to other gram-negative bacteria, we recommend the use of appropriate drugs, especially cephalosporins, in burn hospitals.

  3. Purification Crude Extracellular Protease of Saccharomycopsis fibuligera strain R64 Isolated from Tape’ Indonesian Fermented Food

    OpenAIRE

    L.B. Roostita; W.S. Putranto; C. Charoenchai; E. Wulandari; L.U. Gemilang

    2013-01-01

    The research aims to purify crude extracellular protease produced by Saccharomycopsis fibuligera strain R64 that isolated from Tape' Indonesian fermented food. Saccharomycopsis fibuligera strain R64 was tested on Malt Extract Agar (MEA) that added 3% skim milk powder to discover the proteolytic activity. Extracellular protease purified with ammonium sulfate precipitation, dialysis and gel filtration chromatography with Sephadex G-100. Bradford method used to measure protein content, protease ...

  4. Isolation and characterization of Alicycliphilus denitrificans strain BC, which grows on benzene with chlorate as the electron acceptor

    NARCIS (Netherlands)

    Weelink, S.A.B.; Tan, N.C.G.; Broeke, H. ten; Kieboom, C. van den; Doesburg, W. van; Langenhoff, A.A.M.; Gerritse, J.; Junca, H.; Stams, A.J.M.

    2008-01-01

    A bacterium, strain BC, was isolated from a benzene-degrading chlorate-reducing enrichment culture. Strain BC degrades benzene in conjunction with chlorate reduction. Cells of strain BC are short rods that are 0.6 μm wide and 1 to 2 μm long, are motile, and stain gram negative. Strain BC grows on

  5. Prophages in Enterococcal Isolates from Renal Transplant Recipients: Renal Failure Etiologies Promote Selection of Strains

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    Agnieszka Daca

    2014-01-01

    Full Text Available Infections caused by commensal bacteria may be fatal for the patients under immunosuppressive therapy. This results also from difficulty in identification of high risk strains. Enterococcal infections are increasingly frequent but despite many studies on virulence traits, the difference between commensal and pathogenic strains remains unclear. Prophages are newly described as important elements in competition between strains during colonization, as well as pathogenicity of the strains. Here we evaluate a difference in presence of pp4, pp1, and pp7 prophages and ASA (aggregation substance gene expression in enterococcal isolates from renal transplant recipients (RTx with different etiology of the end-stage renal failure. Prophages sequence was screened by PCR in strains of Enterococcus faecalis isolated from urine and feces of 19 RTx hospitalized at Medical University of Gdansk and 18 healthy volunteers. FLOW-FISH method with use of linear locked nucleic acid (LNA probe was used to assess the ASA gene expression. Additionally, ability of biofilm formation was screened by crystal violet staining method. Presence of prophages was more frequent in fecal isolates from immunocompromised patients than in isolates from healthy volunteers. Additionally, both composition of prophages and ASA gene expression were related to the etiology of renal disease.

  6. SUSCEPTIBILITY OF Listeria monocytogenes STRAINS ISOLATED FROM FOOD TO ANTIMICROBIAL AGENTS

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    E. Zanardi

    2013-02-01

    Full Text Available The objective of this study was to evaluate the susceptibility of 40 L. monocytogenes strains isolated from seafood and processing environments to 19 antibiotics currently used in veterinary and human therapy. Susceptibility tests were performed by the automated system VITEK2. Apart from Penicillin, Ampicillin and Trimethoprim-Sulfamethoxazole, for which clinical breakpoint for Listeria susceptibility testing are defined according to the Clinical and Laboratory Standard Institute (CLSI, in the present study the CLSI criteria for staphylococci were applied. This study shows that isolated L. monocytogenes strains are susceptible to the antibiotics commonly used in veterinary and human listeriosis treatment. Very few strains (7,5% showed a resistance behaviour towards Oxacillin, whereas a variable pattern was showed for Ciprofloxacin and Moxifloxacin. Moreover, an increase in tetracycline resistance, reported by several authors, can not be confirmed in this study, probably due to the different sources of strains isolation. At last, the VITEK2 system represents a rapid and easy-to-use means for antimicrobial susceptibility test of Listeria monocytogenes. In conclusion, because of the increase of antimicrobial resistance showed by L. monocytogenes, a continuous surveillance of emerging antimicrobial resistance among this pathogen is important to ensure effective treatment of human listeriosis. These data can be used for improve background data on antibiotic resistance of strains isolated from food and food environment, even considering the lack of clinical breakpoint provided by the CLSI.

  7. [Activity of imipenem and meropenem against strains of Acinetobacter baumannii isolated from various Chilean hospitals].

    Science.gov (United States)

    Bello, H; Domínguez, M; Sepúlveda, M; Cerda, F; González, G; Mella, S; Zemelman, R

    2000-12-01

    The activity of two carbapenem compounds, imipenem and meropenem, against 447 strains of Acinetobacter baumannii isolated between 1990 and 1998 in different Chilean hospitals was determined. MIC ranges, MIC(50) and MIC(90), were determined by an agar dilution method. Similar antibacterial activities were observed for both antibacterials; however, a slight increase in the MIC(50) of imipenem and meropenem, and in the MIC(90) of meropenem was found among strains isolated from 1997-1998. Although A. baumannii remains susceptible to these antibiotics, the MIC(50) and MIC(90) have increased in recent years.

  8. Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii

    NARCIS (Netherlands)

    Dhaliwal, S.S.; Oberoi, H.S.; Sandhu, S.K.; Nanda, D.; Kumar, D.; Uppal, S.K.

    2011-01-01

    The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from

  9. Comparison of the serum sensitivity of uropathogenic strains of Escherichia coli isolated from different diagnostic groups

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    J. Vraneš,

    2005-02-01

    Full Text Available The bactericidal activity of serum caused by complement system is an important defence mechanism protecting the host organism against infection. The capacity to resist bactericidal activity of normal human serum contributes to the virulence of many gram-negative pathogens. Serum resistance in bacteria has been attributed to their surface components, but exact mechanism of resistance which most likely involves multiple factors is not well understood. In this study, the capacity of Escherichia coli to resist the bactericidal action of serum was examined in 85 clinical isolates obtained from patients with acute pyelonephritis (n=23, acute cystitis (n=22, chronic pyelonephritis (n=22 and asymptomatic bacteriuria (n=18. Serum sensitivity was also examined in relation to the serogroup specificity and expression of the different adhesins of the strains.Bacterial susceptibility to serum killing was measured by assessing regrowth after incubation in serum according to Schiller and Hatch method. The adhesins of E. coli were determined by hemagglutination and inhibition of hemagglutiation, and serotyping was performed on glass slides and confirmed using a mechanized microtechnique.The significant correlation between serum resistance of uropathogenic strains of E. coli and expression of P-fimbriae and O6 serogroup was observed.Theincidence of serum-resistant E. coli strains was significantly higher in strains isolated from urine of patients with acute pyelonephritis, as compared to strains isolated in other diagnostic groups, which is in accordance with higher virulence and invasive potential of these strains.

  10. Isolation of a novel 'atypical' Brucella strain from a bluespotted ribbontail ray (Taeniura lymma).

    Science.gov (United States)

    Eisenberg, Tobias; Riße, Karin; Schauerte, Nicole; Geiger, Christina; Blom, Jochen; Scholz, Holger C

    2017-02-01

    A pleomorphic Gram-negative, motile coccobacillus was isolated from the gills of a wild-caught bluespotted ribbontail ray after its sudden death during quarantine. Strain 141012304 was observed to grow aerobically, to be clearly positive for cytochrome oxidase, catalase, urease and was initially identified as "Brucella melitensis" or "Ochrobactrum anthropi" by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and VITEK2-compact ® , respectively. Affiliation to the genus Brucella was confirmed by bcsp31 and IS711 PCR as well as by Brucella species-specific multiplex PCR, therein displaying a characteristic banding pattern recently described for Brucella strains obtained from amphibian hosts. Likewise, based on recA sequencing, strain 141012304 was found to form a separate lineage, within the so called 'atypical' Brucella, consisting of genetically more distantly related strains. The closest similarity was detected to brucellae, which have recently been isolated from edible bull frogs. Subsequent next generation genome sequencing and phylogenetic analysis confirmed that the ray strain represents a novel Brucella lineage within the atypical group of Brucella and in vicinity to Brucella inopinata and Brucella strain BO2, both isolated from human patients. This is the first report of a natural Brucella infection in a saltwater fish extending the host range of this medically important genus.

  11. Isolation and characterization of new Metschnikowia pulcherrima strains as producers of the antimicrobial pigment pulcherrimin.

    Science.gov (United States)

    Türkel, Sezai; Ener, Beyza

    2009-01-01

    Metschnikowia pulcherrima is a highly effective biocontrol yeast due to its pigment pulcherrimin that accumulates in the cells and in the growth medium. Three different strains of M. pulcherrima were isolated from local grapes. The yeast isolates were characterized on the basis of their biochemical, physiological and ITS1-5.8 s rDNA-ITS2 region. Based on the obtained results, the M. pulcherrima isolates were identified as new strains of M. pulcherrima. Strong antagonistic activities of the M. pulcherrima strains on the human pathogens Proteus vulgaris, Escherichia coli, Candida albicans, Candida parapsilosis, Candida krusei, and Trichosporon mucoides were determined. In addition, antagonistic effects of these M. pulcherrima strains were also tested against Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Trichoderma spp., Paecilomyces spp., and Bipolaris spp. and it was shown that the three different strains of M. pulcherrima also have an antagonistic effect on the growth of these fungal species at different extents. This study showed that all three strains of M. pulcherrima produce the same amount of the pigment pulcherrimin, but their antimicrobial activities on different microorganisms show important variations.

  12. Isolation and Antibiotic Sensitivity of Bacillus thuringinesis Strain From Dump Soil

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    Sarker, D.

    2010-01-01

    Full Text Available Bacillus thuringiensis (or Bt is a commonly used as a pesticide. B. thuringiensis is a naturally-occurring soil bacterium, also occurs naturally in the gut of caterpillars of various types of moths and butterflies, as well as on the dark surface of plants. The xylanase producing bacterial strains were isolated from dump soil. The strains were isolated on xylan agar media and screening was carried out by xylanolysis method. To test the sensitivity of the isolates, ten different antibiotics were used. The strains were tested for resistance to doxycyclin, erythromycin, chloramphenical, cephallaxin, kanamycin, ampicillin, steptomycin, vancomycin, amoxycillin and neomycin. The strains showed sensitive to doxycyclin, erythromycin, chloramphenical, cephallaxin, kanamycin, ampicillin, steptomycin and vancomycin and also showed resistance to amoxycillin and neomycin, when tested by disc diffusion method on nutrient agar plate confirmed by antibiotic spread plate method. The inhibitory effect of B. thuringiensis strains against the test bacteria Bacillus subtilis, Sarcina lutca, Shigella dysenteriae, Shigella sonnei and Pseudomonus aeruginosa examined. It was found that, B. thuringiensis S1, B. thuringiensis S2 and B. thuringiensis S3 strains showed an inhibitory effect on all of the test bacteria.

  13. Assimilation of cholesterol by yeast strains isolated from infant feces and Feta cheese.

    Science.gov (United States)

    Psomas, E I; Fletouris, D J; Litopoulou-Tzanetaki, E; Tzanetakis, N

    2003-11-01

    Eight yeast strains isolated from infant feces and the traditional Greek Feta cheese, selected for their probiotic properties, were tested along with a commercially available strain of Saccharomyces boulardii for their ability to remove cholesterol from a growth medium (yeast extract glucose peptone broth) supplemented with 0.3% Oxgall. The amount of cholesterol removed during 72 h of growth at 37 degrees C revealed significant variations among the yeast strains examined. Two isolates from infant feces, namely Saccharomyces cerevisiae KK1 and Isaatchenkia orientalis KK5.Y.1 and one isolate from Feta cheese, namely S. cerevisiae 832, along with the commercial strain S. boulardii, were able to remove cholesterol from the growth medium after 48 h of incubation at 37 degrees C. However, Saccharomyces strains proved to be able to remove cholesterol even after 24 h of growth at 37 degrees C. The cholesterol removed from the growth medium was not metabolically degraded but was rather assimilated into the yeast cells. The ability to assimilate cholesterol in vitro and to tolerate low pH levels, gastric juice, and bile indicate that S. cerevisiae 832, and especially S. cerevisiae KK1 and I. orientalis KK5.Y.1 (being more bile and gastric juice tolerant because of their human origin) may be promising candidate strains for use as probiotics.

  14. Features of Lactobacillus sakei isolated from Italian sausages: focus on strains from Ventricina del Vastese

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    Carmela Amadoro

    2015-12-01

    Full Text Available In this study bacterial isolates from Ventricina del Vastese sausage, previously identified as Lactobacillus (L. sakei, were characterised genotypically, physiologically and on the basis of some technologically relevant traits. A total of 70 L. sakei isolates from sausages manufactured with spontaneous fermentation in the same producing plant were taken into account. Six genotypic groups were distinguished on the basis of Rep-polymerase chain reaction with the GTG5 primer, some of which were found only in the sausages ripened at temperatures lower than 10°C for the first two months and lower than 16°C for the remaining three months, according to the traditional ripening process. Six strains were selected as representative of the genotypic profiles and further characterised. A high diversity in their fermentation profiles was observed, and different groups were separated on the basis of growth and acidifying capacity in meat extract. None of the strains produced histamine or tyramine in vitro. One strain was able to slightly inhibit Listeria (L. monocytogenes and L. innocua and all six strains were able to slightly inhibit Enterobacteriaceae isolated from Ventricina del Vastese sausages in vitro. Results showed that most L. sakei strains can have a role in improving the safety of low acidity fermented sausages, even though a limited acidifying capacity was observed in a meat-like substrate, and that L. sakei strains able to produce biogenic amines are unlikely to occur in spontaneously fermented meat products.

  15. Decolorization of Methyl Red by Staphylococcus saprophyticus strain AUCASVE3 Isolated from Textile Effluent

    Directory of Open Access Journals (Sweden)

    Azizul Hakim

    2014-01-01

    Full Text Available In this study, an attempt was made to isolate methyl red, an azo dye, decolorizing bacteria from textile effluent. Among three isolates with methyl red decolorizing ability, the potential bacterial isolates, Staphylococcus saprophyticus strain AUCASVE3 was selected for identification and characterization. The organism was found to belong to the genus Staphylococcus and species saprophyticus. The isolate Staphylococcus saprophyticus strain AUCASVE3 could decolorize 600, 700, 800, and 1000 mg/L of methyl red within 24 h and 48 h, respectively, in presence of 1.0 g/L of glucose. This bacterium showed 94 and 97 % methyl red (600 mg L-1 decolorization within 24 and 48 h, respectively. In case of temperature, the ranging between 30-40 �C was fond to be suitable for the decolorization by S. saprophyticus strain AUCASVE3 and a further increase in the temperature drastically affected decolorization activity of bacterial culture. Thus, due to high potential for methyl red decolorization the isolate Staphylococcus saprophyticus strain AUCASVE3 can be used in the biological treatment plant of industrial effluent containing azo dyes.

  16. Screening, identification and characterization of bacteriocins produced by wine-isolated LAB strains.

    Science.gov (United States)

    Ndlovu, B; Schoeman, H; Franz, C M A P; du Toit, M

    2015-04-01

    To screen and identify wine-isolated LAB strains for bacteriocin production, and to identify and characterize bacteriocins. One hundred and fifty-five LAB strains isolated from South African red wines undergoing spontaneous malolactic fermentation were screened for bacteriocin production. Eight isolates were identified to be bacteriocin producers and were identified as Enterococcus faecium. All eight isolates had the same phenotypic and genotypic profiles. The peptides were preliminarily identified as enterocin P using mass spectrometry and further confirmed by PCR-amplifying enterocin P gene. The enterocin activity was inhibited by α-Chymotrypsin, papain and proteinase K treatments. It was heat stable at 37, 60, 80 and 100°C and showed activity over a broad pH range of 2-10. The production of the enterocin followed that of primary metabolite kinetics and, it showed bactericidal effect to some wine spoilage LAB strains. Our study identified the presence of the enterocin-producing Enterococcus in wine. The enterocin was heat stable; with broad pH range and bactericidal effects to sensitive strains. This is one of very few studies that isolated Enterococcus species from wine. It is, however, the first to report presence of bacteriocin-producing Enterococcus in wine fermentation. © 2015 The Society for Applied Microbiology.

  17. Heat shock protein of Mycoplasma salivarium and Mycoplasma orale strains isolated from HIV-seropositive patients.

    Science.gov (United States)

    Chattin-Kacouris, Blanca Rosa; Ishihara, Kazuyuki; Miura, Tadashi; Okuda, Katsuji; Ikeda, Masakazu; Ishikawa, Tatsuya; Rowland, Randal

    2002-11-01

    It has been suggested that infection by some mycoplasma species can act as possible cofactors in the acceleration of immunodeficiency in HIV-infected patients. The present study was designed to examine infections by oral mycoplasma species in HIV-seropositive (HIV(+)) patients. Mycoplasma salivarium and Mycoplasma orale were isolated from 59.5% and 16.7% of 42 HIV(+) patients, respectively. Non-M. salivarium and non-M. orale species were isolated from 40.5% of saliva samples from the HIV(+) group and 20.8% of those from 24 HIV-seronegative (HIV(-)) subjects, respectively. Although the production of superantigen by human peripheral lymphocytes in the isolated mycoplasma species from HIV(+) and HIV(-) subjects was evaluated, none of the examined mycoplasma strains, including ATCC strains of M. salivarium, M. orale, Mycoplasma buccae and Mycoplasma penetrans, were found to produce superantigen. Production of heat shock proteins (HSPs) by isolated mycoplasma strains was examined by immunoblotting using monoclonal antibodies against Helicobacter pylori HSP60. It was found that all the strains of M. salivarium, M. orale, and unidentified mycoplasma species isolated from HIV(+) and HIV(-) groups produced heat shock proteins. HSP production by oral mycoplasma may play a role in the immunomodulation of HIV(+) patients.

  18. Acetobacter strains isolated during the acetification of blueberry (Vaccinium corymbosum L.) wine.

    Science.gov (United States)

    Hidalgo, C; García, D; Romero, J; Mas, A; Torija, M J; Mateo, E

    2013-09-01

    Highbush blueberries (Vaccinium corymbosum L.) are known to have positive health benefits. The production of blueberry vinegar is one method to preserve this seasonal fruit and allow extended consumption. In this study, blueberry wine acetification was performed with naturally occurring micro-organisms and with an inoculated Acetobacter cerevisiae strain. Acetifications were carried out in triplicate using the Schützenbach method. The successful spontaneous processes took up to 66% more time than the processes involving inoculation. The isolation of acetic acid bacteria (AAB) and the analysis of these AAB using molecular methods allowed the identification of the main genotypes responsible of the blueberry acetification. Although the Acet. cerevisiae strain was the predominant strain isolated from the inoculated process samples, Acetobacter pasteurianus was isolated from samples for both processes and was the only species present in the spontaneous acetification samples. To the best of our knowledge, this is the first report describing the identification and variability of AAB isolated during blueberry acetification. The isolated Acet. pasteurianus strains could be used for large-scale blueberry vinegar production or as a starter culture in studies of other vinegar production methods. © 2013 The Society for Applied Microbiology.

  19. Isolation of Escherichia coli 0157:H7 Strain from Fecal Samples of Zoo Animal

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    Aseel Mohammed Hamzah

    2013-01-01

    Full Text Available The isolation and characterization of Escherichia coli O157:H7 strains from 22 out of 174 fecal samples from petting zoo animals representing twenty-two different species (camel, lion, goats, zebra, bear, baboon monkey, Siberian monkey, deer, elk, llama, pony, horses, fox, kangaroo, wolf, porcupine, chickens, tiger, ostrich, hyena, dogs, and wildcats were investigated. One petting Al-Zawraa zoological society of Baghdad was investigated for E. coli O157:H7 over a 16-month period that spanned two summer and two autumn seasons. Variation in the occurrence of E. coli O157:H7-positive petting zoo animals was observed, with animals being culture positive only in the summer months but not in the spring, autumn, or winter. E. coli O157:H7 isolates were distinguished by agglutination with E. coli O157:H7 latex reagent (Oxoid, identified among the isolates, which showed that multiple E. coli strains were isolated from one petting zoo animal, in which a single animal simultaneously shed multiple E. coli strains; E. coli O157:H7 was isolated only by selective enrichment culture of 2 g of petting zoo animal feces. In contrast, strains other than O157:H7 were cultured from feces of petting zoo animals without enrichment.

  20. Variation in susceptibility to benznidazole in isolates derived from Trypanosoma cruzi parental strains

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    Veloso VM

    2001-01-01

    Full Text Available In this work, the susceptibility to benznidazole of two parental Trypanosoma cruzi strains, Colombian and Berenice-78, was compared to isolates obtained from dogs infected with these strains for several years. In order to evaluate the susceptibility to benznidazole two groups of mice were infected with one of five distinct populations isolated from dogs as well as the two parental strains of T. cruzi. The first group was treated with benznidazole during the acute phase and the second remained untreated controls. The animals were considered cured when parasitological and serological tests remained persistently negative. Mice infected with the Colombian strain and its isolates Colombian (A and B did not cure after treatment. On the other hand, all animals infected with Berenice-78 were cured by benznidazole treatment. However, 100%, 50% and 70% of cure rates were observed in animals infected with the isolates Berenice-78 B, C and D, respectively. No significant differences were observed in serological profile of infected control groups, with all animals presenting high antibody levels. However, the ELISA test showed differences in serological patterns between mice inoculated with the different T. cruzi isolates and treated with benznidazole. This variability was dependent on the T. cruzi population used and seemed to be associated with the level of resistance to benznidazole.

  1. Susceptibility of clinical isolates of Bacteroides fragilis group strains to cefoxitin, cefoperazone and ticarcillin/clavulanate

    Directory of Open Access Journals (Sweden)

    PEIXOTO JÚNIOR Arnaldo Aires

    2000-01-01

    Full Text Available A total of 40 strains of the B. fragilis group was isolated from clinical specimens in two hospital centers in Fortaleza from 1993 to 1997. The most frequently isolated species was Bacteroides fragilis (19 strains and most isolates came from intra-abdominal and wound infections. The susceptibility profile was traced for cefoxitin, cefoperazone and ticarcillin-clavulanate by using the agar dilution reference method. All isolates were susceptible to ticarcillin-clavulanate (128/2mug/ml. Resistance rates of 15 and 70% were detected to cefoxitin (64mug/ml and cefoperazone (64mug/ml, respectively. Such regional results permit a better orientation in choosing this group of antibiotics for prophylaxis and therapy especially in relation to cefoxitin, which is frequently used in the hospital centers studied.

  2. Isolation, identification and characterization of an electrogenic microalgae strain.

    Directory of Open Access Journals (Sweden)

    Yicheng Wu

    Full Text Available Extracellular electron transfer involving microbes is important as it closely reflects the ability of cells to communicate with the environment. However, there are few reports on electron transfer mechanisms of pure microalgae and a lack of any model alga to study the transfer processes. In the present study, nine green microalgae species were isolated from wastewater and characterized in terms of their ability to transfer electrons between cells and an electrode. One species showed direct electron transfer via membrane-associated proteins and indirect electron transfer via secreted oxygen. The microalga was identified as Desmodesmus sp. based on phylogenetic analysis and electron microscopy. Electrochemical tests demonstrated that Desmodesmus sp. was able to act as a cathodic microorganism. Stable current densities of -0.24, 35.54 and 170 mA m(-2 were achieved at potentials of +0.2, -0.2 and -0.4 V, respectively, under illumination. Dissolved oxygen concentration measurement showed gradients within the microalgae biofilm: 18.3 mg L(-1 in light decreasing to 4.29 mg L(-1 in the dark. This study diversified the exoelectrogen library and provided a potential model microalga to explore the associated mechanism of extracellular electron transfer.

  3. Antibioresistance of Escherichia coli Strains Isolated from Chicken Colibacillosis in Western Algeria

    OpenAIRE

    HAMMOUDI, Abdelhamid; AGGAD, Hebib

    2014-01-01

    In western Algeria, 251 Escherichia coli isolates were recovered from broilers with clinical signs and lesions of colibacillosis. Serotyping showed that 82% of the isolates belong to one of the following serotypes: O78, O2, and O1. Antibiograms revealed a high level of resistance to oxytetracycline (82%), amoxicilline and ampicilline (47%), trimethoprim-sulfmethoxazole (42%), oxolinic acid and flumequine (31%). However, only a low percentage of strains were resistant to enrofloxacine (6%) and...

  4. Characterization and identification of newly isolated Acinetobacter baumannii strain serdang 1 for phenol removal

    Science.gov (United States)

    Yadzir, Z. H. M.; Shukor, M. Y.; Nazir, M. S.; Abdullah, M. A.

    2012-09-01

    A new indigenous bacterial strain from Malaysian soil contaminated with petroleum waste had been successfully isolated, characterized and identified for phenol removal. The gram negative bacteria showed 98% identity with Acinetobacter baumannii based on Biolog{trade mark, serif} Identification System and the determination of a partial 16S ribosomal RNA (rRNA) sequence. The isolate clustered with species belonging to Acinetobacter clade in a 16S rDNA-based neighbour-joining phylogenetic tree.

  5. Species Identification, Strain Differentiation, and Antifungal Susceptibility of Dermatophyte Species Isolated From Clinically Infected Arabian Horses

    DEFF Research Database (Denmark)

    El Damaty, Hend M; Tartor, Yasmine H; Mahmmod, Yasser Saadeldien Ibrahim

    2017-01-01

    Arabian horses, the eldest equine breeds, have great economic and social significance for its long, unique, and storied history. Molecular characterization of dermatophyte species affecting Arabian horses is a crucial necessity for epidemiologic and therapeutic purposes. The objective of this study...... was to identify and differentiate isolates of dermatophytes isolated from naturally infected Arabian horses at species and strains levels using PCR-restriction fragment length polymorphism (RFLP) and DNA sequencing. Additionally, antifungal susceptibility testing using broth microdilution method was performed...

  6. Serological characterization of Actinobacillus pleuropneumoniae biotype 2 strains isolated from pigs in two Danish herds

    DEFF Research Database (Denmark)

    Nielsen, R.; Andresen, Lars Ole; Plambeck, Tamara

    1997-01-01

    Eight Actinobacillus pleuropneumoniae biotype 2 strains were isolated in pure culture from lungs of pigs originating from two Danish herds with growing and finishing pigs. The antigenic properties were studied by indirect haemagglutination (IHA) and immunodiffusion (ID) tests using soluble surface...... antigens and by enzyme-linked immunosorbent assays (ELISA) using capsular enriched fractions and LPS. In all tests the strains proved antigenically homogeneous and serologically distinct from the known biotype 1 and 2 serotypes. Thus, the strains represent a new serotype which is provisionally proposed...... as biotype 2 serotype 14....

  7. Virulence Factors of Enterococcus faecium and Enterococcus faecalis Strains Isolated from Humans and Pets

    OpenAIRE

    GÜLHAN, Timur; AKSAKAL, Abdulbaki; EKİN, İsmail Hakkı; SAVAŞAN, Serap; BOYNUKARA, Banur

    2006-01-01

    The virulence factors of 146 Enterococcus faecium and 32 Enterococcus faecalis strains isolated from faecal samples of humans, dogs, and cats were investigated. In total, 178 strains were examined by gelatinase (GelE), aggregation substance (AS), cytolysin, and slide haemagglutination tests. The results of detected virulence factors of E. faecium and E. faecalis strains were: GelE: 17.1% vs. 37.5%; AS: 13% vs. 12.5%; cytolysin: 7.5% vs. 12.5%; haemagglutination activities with rabbit erythroc...

  8. Comparison of Biological and Immunological Characterization of Lipopolysaccharides From Brucella abortus RB51 and S19.

    Science.gov (United States)

    Kianmehr, Zahra; Kaboudanian Ardestani, Sussan; Soleimanjahi, Hoorieh; Fotouhi, Fatemeh; Alamian, Saeed; Ahmadian, Shahin

    2015-11-01

    Brucella abortus RB51 is a rough stable mutant strain, which has been widely used as a live vaccine for prevention of brucellosis in cattle instead of B. abortus strain S19. B. abortus lipopolysaccharide (LPS) has unique properties in comparison to other bacterial LPS. In the current study, two types of LPS, smooth (S-LPS) and rough (R-LPS) were purified from B. abortus S19 and RB51, respectively. The aim of this study was to evaluate biological and immunological properties of purified LPS as an immunogenical determinant. Primarily, S19 and RB51 LPS were extracted and purified by two different modifications of the phenol water method. The final purity of LPS was determined by chemical analysis (2-keto-3-deoxyoctonate (KDO), glycan, phosphate and protein content) and different staining methods, following sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). C57BL/6 mice were immunized subcutaneously three times at biweekly intervals with the same amount of purified LPSs. The humoral immunity was evaluated by measuring specific IgG levels and also different cytokine levels, such as IFN-γ, TNF-α, IL-4 and IL-10, were determined for assessing T-cell immune response. Biochemical analysis data and SDS-PAGE profile showed that the chemical nature of S19 LPS is different from RB51 LPS. Both S and R-LPS induce an immune response. T-cell immune response induced by both S and R-LPS had almost the same pattern whereas S19 LPS elicited humoral immunity, which was higher than RB51 LPS. Purified LPS can be considered as a safe adjuvant and can be used as a component in prophylactic and therapeutic vaccines targeting infectious disease, cancer and allergies.

  9. Genetic complementation analysis of two independently isolated hycanthone-resistant strains of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Livia Pica-Mattoccia

    1992-01-01

    Full Text Available The objective of this study is to determine whether various hycanthone resistant strains of schistosomes which have been independently isolated are all affected in the same gene. A strain obtained from a Brazilian patient was compared with a strain of Puerto Rican origin selected in the laboratory. If the mutation conferring resistance involved two different genes, one would expect that the progeny of a cross between the two strains would show complementation, i.e. it would be sensitive to the drug. We have performed such a cross and obtained F1 hybrid worms wich were essentially all resistant, thus suggesting that the mutation conferring resistance in the two strains involves the same gene.

  10. Draft genome sequence of Thermoanaerobacterium sp. strain PSU-2 isolated from thermophilic hydrogen producing reactor.

    Science.gov (United States)

    O-Thong, Sompong; Khongkliang, Peerawat; Mamimin, Chonticha; Singkhala, Apinya; Prasertsan, Poonsuk; Birkeland, Nils-Kåre

    2017-06-01

    Thermoanaerobacterium sp. strain PSU-2 was isolated from thermophilic hydrogen producing reactor and subjected to draft genome sequencing on 454 pyrosequencing and annotated on RAST. The draft genome sequence of strain PSU-2 contains 2,552,497 bases with an estimated G + C content of 35.2%, 2555 CDS, 8 rRNAs and 57 tRNAs. The strain had a number of genes responsible for carbohydrates metabolic, amino acids and derivatives, and protein metabolism of 17.7%, 14.39% and 9.81%, respectively. Strain PSU-2 also had gene responsible for hydrogen biosynthesis as well as the genes related to Ni-Fe hydrogenase. Comparative genomic analysis indicates strain PSU-2 shares about 94% genome sequence similarity with Thermoanaerobacterium xylanolyticum LX-11. The nucleotide sequence of this draft genome was deposited into DDBJ/ENA/GenBank under the accession MSQD00000000.

  11. Mycobacterium eburneum sp. nov., a non-chromogenic, fast-growing strain isolated from sputum.

    Science.gov (United States)

    Nouioui, Imen; Carro, Lorena; Teramoto, Kanae; Igual, José M; Jando, Marlen; Del Carmen Montero-Calasanz, Maria; Sutcliffe, Iain; Sangal, Vartul; Goodfellow, Michael; Klenk, Hans-Peter

    2017-09-01

    A polyphasic study was undertaken to establish the taxonomic position of a non-chromogenic, rapidly growing Mycobacterium strain that had been isolated from sputum. The strain, CECT 8775T, has chemotaxonomic and cultural properties consistent with its classification in the genus Mycobacterium and was distinguished from the type strains of closely related mycobacterial species, notably from Mycobacterium paraense DSM 46749T, its nearest phylogenetic neighbour, based on 16S rRNA, hsp65 and rpoB gene sequence data. These organisms were also distinguished by a broad range of chemotaxonomic and phenotypic features and by a digital DNA-DNA relatedness value of 22.8 %. Consequently, the strain is considered to represent a novel species of Mycobacterium for which the name Mycobacterium eburneum sp. nov is proposed; the type strain is X82T (CECT 8775T=DSM 44358T).

  12. Biodegradation of nicotine by a novel Strain Shinella sp. HZN1 isolated from activated sludge.

    Science.gov (United States)

    Jiang, Hong J; Ma, Yun; Qiu, Guo J; Wu, Fei L; Chen, Sheng L

    2011-01-01

    The nicotine-degrading bacterium HZN1 was isolated from activated sludge and identified as Shinella sp. based on its physiological characteristics and analysis of 16S rDNA gene. Strain HZN1 is capable of using nicotine as the sole carbon source in the mineral salts medium. The optimum temperature and pH for strain HZN1 growth and nicotine degradation were 30°C and 7.0, respectively. It could degrade approximately 100 % of 0.5 g L(-1) of nicotine within 9 h. Three intermediate metabolites were produced by the strain HZN1 and identified as cotinine, myosmine and nicotyrine using gas chromatography-mass spectrometry. This is the first report of nicotine-degrading strain from the genus of Shinella. The results showed that strain HZN1 could be potentially employed in bioremediation of nicotine. Our findings would provide a new insight into the biodegradation of nicotine.

  13. Complete genome sequence of Yersinia pestis strain 91001, an isolate avirulent to humans

    DEFF Research Database (Denmark)

    Song, Yajun; Tong, Zongzhong; Wang, Jin

    2004-01-01

    Genomics provides an unprecedented opportunity to probe in minute detail into the genomes of the world's most deadly pathogenic bacteria- Yersinia pestis. Here we report the complete genome sequence of Y. pestis strain 91001, a human-avirulent strain isolated from the rodent Brandt's vole......-Microtus brandti. The genome of strain 91001 consists of one chromosome and four plasmids (pPCP1, pCD1, pMT1 and pCRY). The 9609-bp pPCP1 plasmid of strain 91001 is almost identical to the counterparts from reference strains (CO92 and KIM). There are 98 genes in the 70,159-bp range of plasmid pCD1. The 106,642-bp...... comparison, we conclude that strain 91001 and other strains isolated from M. brandti might have evolved from ancestral Y. pestis in a different lineage. The large genome fragment deletions in the 91001 chromosome and some pseudogenes may contribute to its unique nonpathogenicity to humans and host...

  14. [Pathogenicity island region of clinical and environmental strains of Vibrio parahaemolyticus, isolated in Chile].

    Science.gov (United States)

    Núñez, Harold; Ulloa, María Teresa; Guerra, Fabiola; Osorio, Carlos G

    2009-02-01

    Most clinical isolates of Vibrio parahaemolyticus produce a major virulence factor known as the thermostable direct hemolysin (TDH). TDH is encoded by the tdh gene which is located in a genomic pathogenicity island (PAI). Most environmental isolates are described as tdh negative. To assess if environmental strains lack the full pathogenicity island or if only the tdh gene is deleted. Thirty eight clinical and 66 environmental strains of Vibrio parahaemolyticus were studied. PAI was characterized by polymerase chain reaction (PCR). The presence of tdhA and tdhS genes, was determined by Southern blot. Fifty three environmental strains (80%) lacked a full PAI when compared with clinical strains. In environmental strains, Southern blot and sequence analysis showed that a genetic region of 80 kilobase pairs including genes from VPA1310 to VPA1396 was missing. These results highlight the genetic dynamism of Vibrio parahaemolyticus pathogenecity island region and suggest that new pathogenic strains could appear by horizontal transfer of the island between toxigenic and non-toxigenic strains.

  15. Diversity of Acetobacter pasteurianus strains isolated from solid-state fermentation of cereal vinegars.

    Science.gov (United States)

    Wu, JiaJia; Gullo, Maria; Chen, FuSheng; Giudici, Paolo

    2010-04-01

    Vinegar production is based on the acetification process by indigenous acetic acid bacteria (AAB). Among vinegar technologies, solid-state fermentation (SSF) processes are widespread in Asian countries to produce vinegar at small-scale. In this study, 21 AAB strains isolated from Chinese cereal vinegars produced by SSF collected in different regions of China were characterized by enterobacterial repetitive intergenic consensus (ERIC)-PCR fingerprinting. Isolates exhibited high degree of phenotypic variability as well as suitable traits for their uses as selected strains in SSF vinegar production (growth modality by superficial biofilm, no production of cellulose, ability to growth on ethanol media). 16S rRNA gene sequencing analysis of representative strains showed that strains of Acetobacter pasteurianus have a close association to cereal vinegars, whereas Gluconacetobacter europaeus population is not favoured. Selection of single or multiple strains culture within A. pasteurianus species was predicted in view of their application in SSF technology. This seems to be the first report showing phenotypic and genetic variability of AAB strains involved in SSF processes. Results can be exploited for the implementation of large-scale SSF processes by selected strains for vinegar production and other innovative biotechnological applications.

  16. Full genomic analysis of a human rotavirus G1P[8] strain isolated in South Korea.

    Science.gov (United States)

    Cho, Min-Kyu; Jheong, Weon-Hwa; Lee, Sung-Geun; Park, Chul Jong; Jung, Kum Hee; Paik, Soon-Young

    2013-01-01

    A rotavirus G1P[8] strain C1-81 was isolated from a 5-month-old female infant admitted to hospital with fever and severe diarrhea in Incheon, South Korea. To investigate its full genomic relatedness and its group, the full genome of strain C1-81 was determined. Based on a full genome classification system, C1-81 was shown to possess the typical Wa-like genotype constellation: G1-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1. On the basis of sequence similarities, the strain was shown to be the closest related strain to contemporary human rotavirus strains with recent strains isolated in Asia. This C1-81 strain showed the highest degree of nucleic acid similarity (98.8% and 97%) to G1 B4633-03 and P[8] (Thai-1604 and Dhaka8-02), respectively. This is the first report that group A rotavirus was analyzed with G1P[8] in South Korea. The study of the complete genome of the virus will help understanding of the evolution of rotavirus. Copyright © 2012 Wiley Periodicals, Inc.

  17. [Adhesion patterns in diffusely adherent Escherichia coli (DAEC) strains isolated from children with and without diarrhea].

    Science.gov (United States)

    Riveros, Maribel; Barletta, Francesca; Cabello, Martín; Durand, David; Mercado, Erik H; Contreras, Carmen; Rivera, Fulton P; Mosquito, Susan; Lluque, Angela; Ochoa, Theresa J

    2011-03-01

    Diffusely adherent E. coli (DAEC) is the sixth recognized group of diarrheagenic E. coli. However, its association with diarrhea remains controversial. Variability in the adherence patterns of clinical strains is unknown. To compare the adherence patterns between strains isolated from children with and without diarrhea. A total of 31 DAEC strains were analyzed, 25 from children with diarrhea and 6 from asymptomatic (control) children, isolated from a cohort study of children under one year of age in the southern districts of Lima. DAEC were identified by PCR (daaD gene). The pattern and adherence score in HEp-2 cell culture were evaluated, Actin polimerization was determined by fluorescence actin staining (FAS) and motility was evaluated by conventional microbiology methods. Diffuse adherence pattern was found in 88% of diarrhea samples and in the total of control strains. The number of bacteria adhered per cell was significantly lower in diarrhea samples (p<0.05). However, actin polymerization was greater in diarrhea samples (60% vs. 17%). Motility test was positive in 60% of the diarrhea samples and in all control samples. Our findings suggest a difference between adherence patterns, actin polymerization and motility between DAEC strains corresponding to diarrhea and control groups. The significance of these results must be confirmed with a bigger number of strains and determining the presence of virulence genes in the strains.

  18. Draft genome assembly of two Pseudoclavibacter helvolus strains, G8 and W3, isolated from slaughterhouse environments

    DEFF Research Database (Denmark)

    Raghupathi, Prem Krishnan; Herschend, Jakob; Røder, Henriette Lyng

    2016-01-01

    We report the draft genome sequences of twoPseudoclavibacter helvolusstrains. Strain G8 was isolated from a meat chopper and strain W3 isolated from the wall of a small slaughterhouse in Denmark. The two annotated genomes are 3.91 Mb and 4.00 Mb in size, respectively.......We report the draft genome sequences of twoPseudoclavibacter helvolusstrains. Strain G8 was isolated from a meat chopper and strain W3 isolated from the wall of a small slaughterhouse in Denmark. The two annotated genomes are 3.91 Mb and 4.00 Mb in size, respectively....

  19. Probiotic Potential of Lactobacillus Strains with Antifungal Activity Isolated from Animal Manure.

    Science.gov (United States)

    Ilavenil, Soundharrajan; Park, Hyung Soo; Vijayakumar, Mayakrishnan; Arasu, Mariadhas Valan; Kim, Da Hye; Ravikumar, Sivanesan; Choi, Ki Choon

    2015-01-01

    The aim of the study was to isolate and characterize the lactic acid bacteria (LAB) from animal manure. Among the thirty LAB strains, four strains, namely, KCC-25, KCC-26, KCC-27, and KCC-28, showed good cell growth and antifungal activity and were selected for further characterization. Biochemical and physiology properties of strains confirmed that the strains are related to the Lactobacillus sp.; further, the 16S rRNA sequencing confirmed 99.99% sequence similarity towards Lactobacillus plantarum. The strains exhibited susceptibility against commonly used antibiotics with negative hemolytic property. Strains KCC-25, KCC-26, KCC-27, and KCC-28 showed strong antifungal activity against Aspergillus fumigatus, Penicillium chrysogenum, Penicillium roqueforti, Botrytis elliptica, and Fusarium oxysporum, respectively. Fermentation studies noted that the strains were able to produce significant amount of lactic, acetic, and succinic acids. Further, the production of extracellular proteolytic and glycolytic enzymes, survival under low pH, bile salts, and gastric juice together with positive bile salt hydrolase (Bsh) activity, cholesterol lowering, cell surface hydrophobicity, and aggregation properties were the strains advantages. Thus, KCC-25, KCC-26, KCC-27, and KCC-28 could have the survival ability in the harsh condition of the digestive system in the gastrointestinal tract. In conclusion, novel L. plantarum KCC-25, KCC-26, KCC-27, and KCC-28 could be considered as potential antimicrobial probiotic strains.

  20. Probiotic Potential of Lactobacillus Strains with Antifungal Activity Isolated from Animal Manure

    Directory of Open Access Journals (Sweden)

    Soundharrajan Ilavenil

    2015-01-01

    Full Text Available The aim of the study was to isolate and characterize the lactic acid bacteria (LAB from animal manure. Among the thirty LAB strains, four strains, namely, KCC-25, KCC-26, KCC-27, and KCC-28, showed good cell growth and antifungal activity and were selected for further characterization. Biochemical and physiology properties of strains confirmed that the strains are related to the Lactobacillus sp.; further, the 16S rRNA sequencing confirmed 99.99% sequence similarity towards Lactobacillus plantarum. The strains exhibited susceptibility against commonly used antibiotics with negative hemolytic property. Strains KCC-25, KCC-26, KCC-27, and KCC-28 showed strong antifungal activity against Aspergillus fumigatus, Penicillium chrysogenum, Penicillium roqueforti, Botrytis elliptica, and Fusarium oxysporum, respectively. Fermentation studies noted that the strains were able to produce significant amount of lactic, acetic, and succinic acids. Further, the production of extracellular proteolytic and glycolytic enzymes, survival under low pH, bile salts, and gastric juice together with positive bile salt hydrolase (Bsh activity, cholesterol lowering, cell surface hydrophobicity, and aggregation properties were the strains advantages. Thus, KCC-25, KCC-26, KCC-27, and KCC-28 could have the survival ability in the harsh condition of the digestive system in the gastrointestinal tract. In conclusion, novel L. plantarum KCC-25, KCC-26, KCC-27, and KCC-28 could be considered as potential antimicrobial probiotic strains.

  1. Molecular characterization of Streptococcus mutans strains isolated from the heart valve of an infective endocarditis patient.

    Science.gov (United States)

    Nemoto, Hirotoshi; Nakano, Kazuhiko; Nomura, Ryota; Ooshima, Takashi

    2008-07-01

    Streptococcus mutans, known to be an aetiological agent of dental caries, is occasionally isolated from patients with infective endocarditis (IE). S. mutans strains with a defect in all three types of glucosyltransferase (GTF) obtained from an infected heart valve extirpated from an IE patient have been reported previously. In this study, molecular analyses of strains detected in heart valve (strain V1) and dental plaque (strain P1) samples taken from the same patient were performed. Complete nucleotide alignments of the gtfB, gtfC and gtfD regions in strains V1 and P1, as well as in the reference strain MT8148, were determined, which revealed the existence of alignments with a high similarity to erythromycin- and spectinomycin-resistance genes in the middle of the gtfB-gtfC and gtfD genes, respectively, of V1. Strain V1 also showed a higher MIC for these two antibiotics compared with strain P1. Next, primers to detect the specific sequences of the antibiotic-resistance genes in strain V1 were constructed and PCR amplification was performed with template DNA from dental plaque and infected valve tissue samples taken from the patient. Attenuated expression of GTFs in V1 caused a significantly lower susceptibility to phagocytosis by human polymorphonuclear leukocytes compared with the reference strain. These results suggest that the blood isolate V1 found in the oral cavity invaded and survived in the bloodstream for a long duration and that this was related to its virulence in IE in our patient.

  2. Screening and evaluation of polyhydroxybutyrate-producing strains from indigenous isolate Cupriavidus taiwanensis strains.

    Science.gov (United States)

    Wei, Yu-Hong; Chen, Wei-Chuan; Huang, Chin-Kuei; Wu, Ho-Shing; Sun, Yi-Ming; Lo, Chi-Wei; Janarthanan, Om-Murugan

    2011-01-05

    Polyhydroxyalkanoate (PHA) is a biodegradable material with many potential biomedical applications, including medical implants and drug delivery. This study developed a system for screening production strains in order to optimize PHA production in Cupriavidus taiwanensis 184, 185, 186, 187, 204, 208, 209 and Pseudomona oleovorans ATCC 29347. In this study, Sudan black B staining, Infrared (IR) and Gas Chromatography (GC) analysis indicated that the best strain for PHA synthesis is C. taiwanensis 184, which obtains polyhydroxybutyrate (PHB). Cultivation of C. taiwanensis 184 under a pH of 7.0, at 30 °C, and at an agitation rate of 200 rpm, obtained a PHB content of 10% and PHB production of 0.14 g/L. The carbon and nitrogen types selected for analysis of PHB production by C. taiwanensis 184 were gluconic acid and NH(4)Cl, respectively. Optimal carbon/nitrogen ratio for PHB production was also determined. This study demonstrated a PHB content of 58.81% and a PHB production of 2.44 g/L when the carbon/nitrogen ratio of 8/1 was selected for C. taiwanensis 184. A two-stage fermentation strategy significantly enhanced PHB content and PHB production. Under a two-stage fermentation strategy with nutrient-limited conditions, C. taiwanensis 184 obtained a PHB content of 72% and a PHB concentration of 7 g/L. Finally, experimental results confirmed that optimizing the growth medium and fermentation conditions for cultivating the indigenous C. taiwanensis 184 strain substantially elevated PHB content from 10% to 72% and PHB production from 0.14 g/L to 7 g/L, respectively.

  3. Screening and Evaluation of Polyhydroxybutyrate-Producing Strains from Indigenous Isolate Cupriavidus taiwanensis Strains

    Directory of Open Access Journals (Sweden)

    Om-Murugan Janarthanan

    2011-01-01

    Full Text Available Polyhydroxyalkanoate (PHA is a biodegradable material with many potential biomedical applications, including medical implants and drug delivery. This study developed a system for screening production strains in order to optimize PHA production in Cupriavidus taiwanensis 184, 185, 186, 187, 204, 208, 209 and Pseudomona oleovorans ATCC 29347. In this study, Sudan black B staining, Infrared (IR and Gas Chromatography (GC analysis indicated that the best strain for PHA synthesis is C. taiwanensis 184, which obtains polyhydroxybutyrate (PHB. Cultivation of C. taiwanensis 184 under a pH of 7.0, at 30 °C, and at an agitation rate of 200 rpm, obtained a PHB content of 10% and PHB production of 0.14 g/L. The carbon and nitrogen types selected for analysis of PHB production by C. taiwanensis 184 were gluconic acid and NH4Cl, respectively. Optimal carbon/nitrogen ratio for PHB production was also determined. This study demonstrated a PHB content of 58.81% and a PHB production of 2.44 g/L when the carbon/nitrogen ratio of 8/1 was selected for C. taiwanensis 184. A two‑stage fermentation strategy significantly enhanced PHB content and PHB production. Under a two-stage fermentation strategy with nutrient‑limited conditions, C. taiwanensis 184 obtained a PHB content of 72% and a PHB concentration of 7 g/L. Finally, experimental results confirmed that optimizing the growth medium and fermentation conditions for cultivating the indigenous C. taiwanensis 184 strain substantially elevated PHB content from 10% to 72% and PHB production from 0.14 g/L to 7 g/L, respectively.

  4. Screening and Evaluation of Polyhydroxybutyrate-Producing Strains from Indigenous Isolate Cupriavidus taiwanensis Strains

    Science.gov (United States)

    Wei, Yu-Hong; Chen, Wei-Chuan; Huang, Chin-Kuei; Wu, Ho-Shing; Sun, Yi-Ming; Lo, Chi-Wei; Janarthanan, Om-Murugan

    2011-01-01

    Polyhydroxyalkanoate (PHA) is a biodegradable material with many potential biomedical applications, including medical implants and drug delivery. This study developed a system for screening production strains in order to optimize PHA production in Cupriavidus taiwanensis 184, 185, 186, 187, 204, 208, 209 and Pseudomona oleovorans ATCC 29347. In this study, Sudan black B staining, Infrared (IR) and Gas Chromatography (GC) analysis indicated that the best strain for PHA synthesis is C. taiwanensis 184, which obtains polyhydroxybutyrate (PHB). Cultivation of C. taiwanensis 184 under a pH of 7.0, at 30 °C, and at an agitation rate of 200 rpm, obtained a PHB content of 10% and PHB production of 0.14 g/L. The carbon and nitrogen types selected for analysis of PHB production by C. taiwanensis 184 were gluconic acid and NH4Cl, respectively. Optimal carbon/nitrogen ratio for PHB production was also determined. This study demonstrated a PHB content of 58.81% and a PHB production of 2.44 g/L when the carbon/nitrogen ratio of 8/1 was selected for C. taiwanensis 184. A two-stage fermentation strategy significantly enhanced PHB content and PHB production. Under a two-stage fermentation strategy with nutrient-limited conditions, C. taiwanensis 184 obtained a PHB content of 72% and a PHB concentration of 7 g/L. Finally, experimental results confirmed that optimizing the growth medium and fermentation conditions for cultivating the indigenous C. taiwanensis 184 strain substantially elevated PHB content from 10% to 72% and PHB production from 0.14 g/L to 7 g/L, respectively. PMID:21339985

  5. Characterization of a Bacillus thuringiensis strain collection isolated from diverse Costa Rican natural ecosystems.

    Science.gov (United States)

    Arrieta, Glen; Espinoza, Ana M

    2006-03-01

    Costa Rican natural ecosystems are among the most diverse in the world. For this reason, we isolated strains of the entomopathogenic bacteria Bacillus thuringiensis (Bt) to determine their diversity, distribution and abundance. A total of 146 Bt strains were obtained from environmental samples collected from diverse natural ecosystems and life zones of Costa Rica. We recovered Bt strains from 71%, 63%, 61% and 54% of soil samples, fresh leaves, other substrates and leaf litter respectively. Bt was isolated in 65% of the samples collected in the humid tropical forest in national parks (Braulio Carrillo, Gandoca Manzanillo, Sierpe, Hitoy Cerere, and Cahuita), and in 59% of the samples collected in the dry tropical forest (Parque Nacional Marino las Baulas, Palo Verde and Santa Rosa). In the very humid tropical forest (Tortuguero) Bt was isolated in 75% of the samples and in the very humid tropical forest transition perhumid (Carara) it was found in 69% of the samples. The strains exhibit a diverse number, size and morphology of parasporal inclusion bodies: irregular (47%), oval (20%), bipyramidal (3%), bipyramidal and cubic (1%), bipyramidal, oval and irregular (5%) and bipyramidal, oval and cubic crystals (2%). Strains isolated from Braulio Carrillo, Tortuguero and Cahuita, presented predominantly irregular crystals. On the other hand, more than 60% of the isolates from Térraba-Sierpe and Hitoy-Cerere had medium oval crystals. Strains from Gandoca-Manzanillo, Palo Verde and Carara presented mainly combinations of oval and irregular crystals. Nevertheless, the greatest diversity in crystal morphology was observed in those from Santa Rosa, Llanos del Rio Medio Queso and Parque Marino las Baulas. Protein analyses of the crystal-spore preparations showed delta-endotoxin with diverse electrophoretic patterns, with molecular weights in the range of 20 to 160 kDa. Fifty six percent of the strains amplified with the cry2 primer, 54% with vip3, 20% with cry1, 9% with cry3

  6. Isolation of a strain of Pseudomonas putida capable of metabolizing anionic detergent sodium dodecyl sulfate (SDS).

    Science.gov (United States)

    Chaturvedi, V; Kumar, A

    2011-03-01

    Sodium Dodecyl Sulfate (SDS) is one of the most widely used anionic detergents. The present study deals with isolation and identification of SDS-degrading bacteria from a detergent contaminated pond situated in Varanasi city, India. Employing enrichment technique in minimal medium (PBM), SDS-degrading bacteria were isolated from pond water sample. Rate of degradation of SDS was studied in liquid PBM and also degradation of different concentrations of SDS was also studied to find out maximum concentration of SDS degraded by the potent isolates. Alkyl sulfatase activity (key enzyme in SDS degradation) was estimated in crude cell extracts and multiplicity of alkyl sulfatase was studied by Native PAGE Zymography. The potent isolate was identified by 16S rRNA sequence analysis. Using enrichment technique in minimal medium containing SDS as a sole carbon source, initially three SDS degrading isolates were recovered. However, only one isolate, SP3, was found to be an efficient degrader of SDS. It was observed that this strain could completely metabolize 0.1% SDS in 16 h, 0.2% SDS in 20 h and 0.3% SDS in 24 h of incubation. Specific activity of alkyl sulfatase was 0.087±0.004 µmol SDS/mg protein/min and Native PAGE Zymography showed presence of alkyl sulfatase of Rf value of 0.21. This isolate was identified as Pseudomonas putida strain SP3. This is the report of isolation of SDS-degrading strain of P. putida, which shows high rate of SDS degradation and can degrade up to 0.3% SDS. It appears that this isolate can be exploited for bioremediation of this detergent from water systems.

  7. Virulence factors and genetic variability of Staphylococcus aureus strains isolated from raw sheep's milk cheese.

    Science.gov (United States)

    Spanu, Vincenzo; Spanu, Carlo; Virdis, Salvatore; Cossu, Francesca; Scarano, Christian; De Santis, Enrico Pietro Luigi

    2012-02-01

    Contamination of dairy products with Staphylococcus aureus can be of animal or human origin. The host pathogen relationship is an important factor determining genetic polymorphism of the strains and their potential virulence. The aim of the present study was to carry out an extensive characterization of virulence factors and to study the genetic variability of S. aureus strains isolated from raw ewe's milk cheese. A total of 100 S. aureus strains isolated from cheese samples produced in 10 artisan cheese factories were analyzed for the presence of enterotoxins (sea-see) and enterotoxins-like genes (seh, sek, sel, sem, seo, sep), leukocidins, exfoliatins, haemolysins, toxic shock syndrome toxin 1 (TSST-1) and the accessory gene regulator alleles (agr). Strains were also typed using pulsed-field gel electrophoresis (PFGE). AMOVA analysis carried out on PFGE and PCR data showed that the major component explaining genetic distance between strains was the dairy of origin. Of the total isolates 81% had a pathogenicity profile ascribable to "animal" biovar while 16% could be related to "human" biovar. The biovar allowed to estimate the most likely origin of the contamination. Minimum inhibitory concentrations (MICs) of nine antimicrobial agents and the presence of the corresponding genes coding for antibiotic resistance was also investigated. 18 strains carrying blaZ gene showed resistance to ampicillin and penicillin and 6 strains carrying tetM gene were resistant to tetracycline. The presence of mecA gene and methicillin resistance, typical of strains of human origin, was never detected. The results obtained in the present study confirm that S. aureus contamination in artisan cheese production is mainly of animal origin. Copyright © 2011. Published by Elsevier B.V.

  8. Comparative effectiveness of isolation techniques for contemporary Influenza A virus strains circulating in exhibition swine.

    Science.gov (United States)

    Bowman, Andrew S; Nelson, Sarah W; Edwards, Jody L; Hofer, Christian C; Nolting, Jacqueline M; Davis, Ian C; Slemons, Richard D

    2013-01-01

    The current study sought to compare the effectiveness of 2 virus isolation methods for the recovery of contemporary Influenza A virus (FLUAV) strains circulating in swine at agricultural exhibitions. Following the emergence of the influenza A (H1N1)pdm09 virus, increased surveillance of FLUAV strains among swine was recommended for early detection of emerging strains that threaten animal and human health. The increase in genetic drift and genomic reassortment among FLUAV strains infecting swine since 1998 necessitates that detection protocols be periodically validated for contemporary FLUAV strains. During 2009, nasal swabs were collected from 221 clinically healthy pigs at 12 agricultural exhibitions in Ohio. Nasal swabs were tested in parallel for the presence of FLUAV strains using 3 methodologies: 2 passages through Madin-Darby canine kidney (MDCK) cells adapted to serum-free medium (SFM), 2 passages through embryonated chicken eggs (ECEs), and real-time reverse transcription polymerase chain reaction (real-time RT-PCR). Of the 221 samples, 40 (18.1%) were positive for FLUAV recovery in MDCK cell culture and 13 (5.9%) were positive in ECEs (P = 0.015). All samples positive in ECEs were also positive in MDCK cell culture. MDCK cell culture virus isolation results were in perfect agreement with results of the real-time RT-PCR. Hemagglutinin and neuraminidase combinations of the recovered isolates were H1N2 and H3N2, which were consistent with FLUAV strains circulating in U.S. pigs. Effectiveness and cost savings justify the use of SFM-adapted MDCK cell culture over ECEs for the recovery of contemporary FLUAV strains from exhibition swine.

  9. Phylogenomics of Brazilian epidemic isolates of Mycobacterium abscessus subsp. bolletii reveals relationships of global outbreak strains

    Science.gov (United States)

    Davidson, Rebecca M.; Hasan, Nabeeh A.; de Moura, Vinicius Calado Nogueira; Duarte, Rafael Silva; Jackson, Mary; Strong, Michael

    2013-01-01

    Rapidly growing, non-tuberculous mycobacteria (NTM) in the Mycobacterium abscessus (MAB) species are emerging pathogens that cause various diseases including skin and respiratory infections. The species has undergone recent taxonomic nomenclature refinement, and is currently recognized as two subspecies, M. abscessus subsp. abscessus (MAB-A) and M. abscessus subsp. bolletii (MAB-B). The recently reported outbreaks of MAB-B in surgical patients in Brazil from 2004 to 2009 and in cystic fibrosis patients in the United Kingdom (UK) in 2006 to 2012 underscore the need to investigate the genetic diversity of clinical MAB strains. To this end, we sequenced the genomes of two Brazilian MAB-B epidemic isolates (CRM-0019 and CRM-0020) derived from an outbreak of skin infections in Rio de Janeiro, two unrelated MAB strains from patients with pulmonary infections in the United States (US) (NJH8 and NJH11) and one type MAB-B strain (CCUG 48898) and compared them to 25 publically available genomes of globally diverse MAB strains. Genome-wide analyses of 27,598 core genome single nucleotide polymorphisms (SNPs) revealed that the two Brazilian derived CRM strains are nearly indistinguishable from one another and are more closely related to UK outbreak isolates infecting CF patients than to strains from the US, Malaysia or France. Comparative genomic analyses of six closely related outbreak strains revealed geographic-specific large-scale insertion/deletion variation that corresponds to bacteriophage insertions and recombination hotspots. Our study integrates new genome sequence data with existing genomic information to explore the global diversity of infectious M. abscessus isolates and to compare clinically relevant outbreak strains from different continents. PMID:24055961

  10. Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status.

    Science.gov (United States)

    Ewers, Christa; Lübke-Becker, Antina; Bethe, Astrid; Kiebling, Sabine; Filter, Matthias; Wieler, Lothar H

    2006-05-31

    To learn more about the molecular biology of Pasteurella multocida 289 strains isolated from various clinically healthy and diseased hosts were examined for capsule biosynthesis genes (capA, B, D, E, and F) and 14 virulence associated genes by PCR and DNA-DNA-hybridization. As expected, capsule type A strains were highly adapted to bovines (92.3%) and poultry (85.7%) while we mainly found capA (34.9%)- and capD (58.1%)-positive strains in swine. A noticeable amount of capD-positive strains also originated from small ruminants (34.9%) and capF was detected in wild type strains from diseased cattle (2.2%) and cats (7.4%). None of the isolates harboured capE, while capB was exclusively found in all strains from buffaloes. Nearly all isolates showed a combination of genes encoding outer membrane proteins, colonization factors, iron aquisition factors and superoxid-dismutases without any clue for host specificity. In contrast, the transferrin binding protein encoding gene tbpA (31.5%) was limited to ruminant strains and only 37.0% of all P. multocida strains harboured pfhA, coding for a filamentous hemagglutinin, supposed to be a putative adhesion- und serum resistance factor. PfhA revealed a strong positive association to the outcome of disease in bovine hosts and in combination with toxA to that in swine. The dermonecrotoxin encoding toxA, present in 12.5% of all strains, was detected in isolates from swine, small ruminants, cattle, and poultry. A significant association to the disease status, however, was only existent in swine, although with 66.7% we found a notably high prevalence of the toxin gene among strains from small ruminants. The genes toxA, tbpA and pfhA as well as capsule biosynthesis genes are supposed to be important epidemiological marker genes for characterizing P. multocida field strains.

  11. Characterization of Methylobacterium strains isolated from the phyllosphere and description of Methylobacterium longum sp. nov.

    Science.gov (United States)

    Knief, Claudia; Dengler, Vanina; Bodelier, Paul L E; Vorholt, Julia A

    2012-01-01

    Methylobacterium strains are abundantly found in the phyllosphere of plants. Morphological, physiological and chemotaxonomical properties of 12 previously isolated strains were analyzed in order to obtain a more detailed overview of the characteristics of phyllosphere colonizing Methylobacterium strains. All strains showed the typical properties of the genus Methylobacterium, including pink pigmentation, facultative methylotrophy, a fatty acid profile dominated by C18:1 ω7c, and a high G+C content of 65 mol % or more. However, some strains showed only weak growth on methanol and pigmentation varied from pale pink to red. Strains grew best under mesophilic, neutrophilic conditions and low salt (≤1%) concentrations, but variation was seen with respect to the temperature and pH range under which growth occurred. Likewise, differences were seen with respect to carbon source utilization. Some strains were versatile and utilized diverse organic acids, amino acids and sugars, while others could only metabolize a restricted number of organic acids. The strains that were most distinct from existing type strains based on 16S rRNA gene sequence analysis were selected for DNA-DNA hybridization experiments to analyze whether they are sufficiently different at the genomic level from existing type strains to justify their classification as new species. This resulted in the delineation of strain 440 and its description as Methylobacterium longum sp. nov. strain 440 (=DSM 23933(T) = CECT 7806(T)). A main characteristic of this species is the formation of relatively long rods compared to other Methylobacterium species.

  12. Clinical analysis of bacterial strain profiles isolated from urinary tract infections: A 30-year study.

    Science.gov (United States)

    Wada, Koichiro; Uehara, Shinya; Yamamoto, Masumi; Sadahira, Takuya; Mitsuhata, Ritsuko; Araki, Motoo; Kobayashi, Yasuyuki; Ishii, Ayano; Kariyama, Reiko; Watanabe, Toyohiko; Nasu, Yasutomo; Kumon, Hiromi

    2016-07-01

    We analyzed bacterial strains isolated from urine samples of patients with urinary tract infections (UTI) at Okayama University Hospital over a 30-year period to characterize trends in species and antimicrobial susceptibilities. Clinical isolates were collected from in- and out-patients with pyuria and bacteriuria who were treated between 1984 and 2014 (one episode per patient and plural isolates were counted in polymicrobial infection). We examined these isolates to identify pathogens and tested for antimicrobial susceptibility. Isolates from complicated UTI over a 30-year period revealed Pseudomonas aeruginosa (P. aeruginosa) was the most frequently isolated in the first decade (1984-1994), MRSA in the second decade (1995-2004), and Escherichia coli (E. coli) in the latest decade (2005-2014). In uncomplicated UTI examined over 20 years, E. coli was the most frequently isolated species accounting for 47-94% of isolates. Fluoroquinolone (FQs)-insusceptible E. coli were first isolated in 1994 and increased to about 35% in 2013 in patients with complicated UTI. Complicated UTI involving P. aeruginosa and MRSA decreased over the last 10 years. Our data suggest that several factors such as shorter hospitalizations, shorter indwelling catheter use, and appropriate antimicrobial use has decreased colonization of P. aeruginosa and MRSA with relative increases in isolation of E. coli including FQs-insusceptible strains. We must continue our surveillance of antimicrobial-resistant bacteria isolated from urine samples and evaluate antibiograms, since their persistence in the urinary tract would be problematic. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  13. Isolation and characterization of Bacillus thuringiensis strains native to Assam soil of North East India.

    Science.gov (United States)

    Rabha, Mihir; Sharma, Shaswati; Acharjee, Sumita; Sarmah, Bidyut Kumar

    2017-10-01

    We have identified both crystalliferous and acrystalliferous Bt isolates from the Assam soil of North East India for the first time. A total of 301 Bacillus type colonies were selected based on their appearance and colony morphology. Out of these colonies, 42 isolates had characteristics similar to Bt isolates on MYP (Mannitol Egg Yolk Polymyxin) agar base medium. The ERIC-PCR and 16S rDNA analyses confirmed that 42 isolates are Bacillus thuringiensis. Phase contrast microscopy showed that 37 isolates produced crystal endospore during the sporulation phase and 5 acrystalliferous isolates were also found. Amplification of cry gene was carried out using general Cry primers along with one cry2 gene specific primer. Out of 42 isolates, 50% of the isolates showed presence of cry2 gene followed by cry9 (40.47) and cry1 (40.47). Moreover, 21.42% of isolates showed the presence of more than one cry genes. We also screened these isolates for the possibility of having new Bt genes using universal primer and found two strains having a new type of Cry1I gene with 82 and 85% similarities with the available Cry1I gene sequences. Thus, these new types of Bt gene could be useful for Bt-based bioformulations and generation of transgenic plants.

  14. Genome sequencing and analysis of a highly virulent Vibrio parahaemolyticus strain isolated from the marine environment

    Science.gov (United States)

    Parks, M. C.; Moreno, E.

    2016-02-01

    Vibrio parahaemolyticus [Vp] is a Gram-negative bacterium and a natural inhabitant of coastal marine ecosystems worldwide. Vp is also a coincidental pathogen of humans. Virulent strains are commonly identified by the presence of the thermostable direct (tdh) or tdh-related (trh) hemolysin genes. However, virulence is multifaceted and many clinical Vp isolates do not carry tdh or trh. In this study, we sequenced and assembled the draft genome of a tdh- and trh-negative environmental isolate (805) shown previously to be highly virulent in zebrafish. To investigate potential mechanisms of virulence, we compared 805 to the clinical V. parahaemolyticus type strain (RIMD2210633). Pairwise comparison revealed the presence of multiple genomic regions including an IncF conjugative pilus (1.3 Kb) and a colicin V plasmid (1.49 Kb). These features are homologous to genomic regions present in clinical V. vulnificus and V. cholerae strains. Genome comparison also revealed the presence of five toxin-antitoxin systems. Isolate 805 likely attained these new features through the lateral acquisition of mobile genomic material - a hypothesis supported by the aberrant GC content of these regions. Colicin V plasmids are a diverse group of IncF plasmids found in invasive bacterial strains. Similarly, an abundance of toxin-antitoxin systems have been linked to virulence in Gram-negative bacteria. Current efforts are focused on characterizing 142 coding features present in 805 but absent from the type strain.

  15. Detection of enterotoxins and genotyping of Staphylococcus aureus strains isolated from Isfahan Educational Hospital, Iran

    Directory of Open Access Journals (Sweden)

    Seyed Asghar Havaei

    2017-10-01

    Full Text Available Background and aims: Staphylococcus aureus is known as one of the most important nosocomial pathogens, which may lead to several infections. The aim of this study was determining the enterotoxins A, C, and TSST-1 and molecular characterization of S. aureus strains with PFGE and MLST typing methods. Materials and methods: In the present study during the sixmonths sampling, fifty S. aureus strains were isolated from patients admitted to Al-Zahra university hospital. Antimicrobial susceptibility testing, Multiplex PCR for detection of enterotoxin A, C and TSST-1, pulse field gel electrophoresis (PFGE and multilocus sequence typing (MLST were used for molecular typing. Results: In antibiogram the highest and lowest percentage of resistance was belonged to tetracycline and rifampin respectively. Multiplex PCR indicated that 30% of the strains harbored sea and 34% harbored sec genes. However, only 4% of our collected isolates had tsst gene. In PFGE method analysis on all S. aureus strains, a total of 19 different patterns were identified. Nine various sequence types in 27 selected S. aureus isolates were identified by MLST. Conclusions: Present study indicates a possible higher variability among our S. aureus strains by two different molecular typing methods; nevertheless four main common types (CT1, CT7, CT9, and CT11 with at least one toxin genes were determined.

  16. Isolation and Fatty Acid Profile of Selected Microalgae Strains from the Red Sea for Biofuel Production

    Directory of Open Access Journals (Sweden)

    Khalid M. Abu-Salah

    2013-05-01

    Full Text Available The isolation of lipid-rich autochthonous strains of microalgae is a crucial stage for the development of a microalgae-based biofuel production plant, as these microalgae already have the necessary adaptations to withstand competition, predation and the temperatures observed at each production site. This is particularly important in extreme climates such as in Saudi Arabia. Resorting to fluorescence activated cell sorting (FACS we screened for and isolated several microalgal strains from samples collected from the Red Sea. Relying on the fluorescence of BODIPY 505/515 (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diazasindacene and growth performance, four promising candidates were identified and the total lipid content and fatty acid profile was assessed for biofuels production. Selected isolates were classified as chlorophytes, belonging to three different genera: Picochlorum, Nannochloris and Desmochloris. The lipid contents were assessed microscopically by means of BODIPY 505/515-associated fluorescence to detect intracellular lipid bodies, which revealed several lipid drops in all selected strains. This result was confirmed by lipid gravimetric determination, which demonstrated that all strains under study presented inner cell lipid contents ranging from 20% to 25% of the biomass dry weight. Furthermore, the fatty acid methyl esters profile of all strains seems ideal for biodiesel production due to a low degree of polyunsaturated fatty acid methyl esters and high amount of palmitic and oleic acids.

  17. Antimicrobial Activity of Monoramnholipids Produced by Bacterial Strains Isolated from the Ross Sea (Antarctica

    Directory of Open Access Journals (Sweden)

    Pietro Tedesco

    2016-04-01

    Full Text Available Microorganisms living in extreme environments represent a huge reservoir of novel antimicrobial compounds and possibly of novel chemical families. Antarctica is one of the most extraordinary places on Earth and exhibits many distinctive features. Antarctic microorganisms are well known producers of valuable secondary metabolites. Specifically, several Antarctic strains have been reported to inhibit opportunistic human pathogens strains belonging to Burkholderia cepacia complex (Bcc. Herein, we applied a biodiscovery pipeline for the identification of anti-Bcc compounds. Antarctic sub-sea sediments were collected from the Ross Sea, and used to isolate 25 microorganisms, which were phylogenetically affiliated to three bacterial genera (Psychrobacter, Arthrobacter, and Pseudomonas via sequencing and analysis of 16S rRNA genes. They were then subjected to a primary cell-based screening to determine their bioactivity against Bcc strains. Positive isolates were used to produce crude extracts from microbial spent culture media, to perform the secondary screening. Strain Pseudomonas BNT1 was then selected for bioassay-guided purification employing SPE and HPLC. Finally, LC-MS and NMR structurally resolved the purified bioactive compounds. With this strategy, we achieved the isolation of three rhamnolipids, two of which were new, endowed with high (MIC < 1 μg/mL and unreported antimicrobial activity against Bcc strains.

  18. Isolation and identification of bacterial strain I33M producing milk ...

    African Journals Online (AJOL)

    Igbigbi

    Full Length Research Paper. Isolation and identification of bacterial strain I33M producing milk-clotting enzyme: Optimization of culture parameters using response surface. Talantikite-Kellil, S1, Nouani, A. 1,2*, Saunier, M3, Gillmann, L3, Brahmi, A4 and Fazouane, F1. 1Food Technology Laboratory, University of Boumerdes ...

  19. Virulence potential of Enterococcus gallinarum strains isolated from selected Nigerian traditional fermented foods

    Directory of Open Access Journals (Sweden)

    IYABO C. OLADIPO

    2014-08-01

    Full Text Available Five Enterococcus isolates from some Nigerian traditional fermented foods were identified as Enterococcus gallinarum by using phenotypic and genotypic tests. Safety properties such as antibiotic susceptibility, virulence gene detection, haemolysin, gelatinase and bacteriocin production were determined using standard methods. There was no resistance to clinically relevant antibiotics. Virulence gene for collagen binding antigen and aggregation substance were detected in 60% of the E. gallinarum strains; while surface adhesin was detected in 20%, but none of the strains had cytolysin activator and gelatinase. Phenotype characterizations of the E. gallinarum isolates indicated that none of the isolates produced haemolysin and gelatinase. Enterococcus gallinarum C103 and U82 had no antimicrobial activity against all the selected bacteria pathogens while E. gallinarum W184, T71 and W21 were active against some of the indicator bacteria pathogens. Only E. gallinarum T71 and W21 showed broad spectra of antimicrobial activity. Combination of virulence factors did not appear in these food isolates. Therefore, these strains particularly the two strains with high spectra of antimicrobial activity could be exploited as functional starters in foods.

  20. Complete Genome Sequence of Bacillus amyloliquefaciens Strain BH072, Isolated from Honey

    NARCIS (Netherlands)

    Zhao, Xin; de Jong, Anne; Zhou, Zhijiang; Kuipers, Oscar P

    2015-01-01

    The genome of Bacillus amyloliquefaciens strain BH072, isolated from a honey sample and showing strong antimicrobial activity against plant pathogens, is 4.07 Mb and harbors 3,785 coding sequences (CDS). Several gene clusters for nonribosomal synthesis of antimicrobial peptides and a complete gene

  1. Genome sequence of Pseudomonas sp. strain PAMC 25886, isolated from alpine glacial cryoconite.

    Science.gov (United States)

    Shin, Seung Chul; Kim, Su Jin; Hong, Soon Gyu; Ahn, Do Hwan; Lee, Yung Mi; Lee, Hyoungseok; Lee, Jungeun; Park, Hyun

    2012-04-01

    Pseudomonas spp. have shown characteristics of efficiently metabolizing environmental pollutants and also producing exopolysaccharides known as biofilms. Here we present the draft genome sequence of Pseudomonas sp. strain PAMC 25886, which was isolated from glacier cryoconite in the Alps mountain permafrost region and which may provide further insight into biodegradative and/or biofilm-producing mechanisms in a cold environment.

  2. Genome comparison and physiological characterization of eight Streptococcus thermophilus strains isolated from Italian dairy products

    DEFF Research Database (Denmark)

    Vendramin, Veronica; Treu, Laura; Campanaro, Stefano

    2017-01-01

    Eight Streptococcus thermophilus strains of dairy origin isolated in Italy were chosen to investigate autochthonous bacterial diversity in this important technological species. In the present study a comparative analysis of all the 17 S. thermophilus genomes publicly available was performed to id...

  3. Draft Genome Sequence of Taylorella equigenitalis Strain MCE529, Isolated from a Belgian Warmblood Horse.

    Science.gov (United States)

    Hébert, Laurent; Touzain, Fabrice; de Boisséson, Claire; Breuil, Marie-France; Duquesne, Fabien; Laugier, Claire; Blanchard, Yannick; Petry, Sandrine

    2014-11-26

    Taylorella equigenitalis is the causative agent of contagious equine metritis (CEM), a sexually transmitted infection of horses. We herein report the genome sequence of T. equigenitalis strain MCE529, isolated in 2009 from the urethral fossa of a 15-year-old Belgian Warmblood horse in France. Copyright © 2014 Hébert et al.

  4. Chlamydia pneumoniae Clinical Isolate from Gingival Crevicular Fluid: A Potential Atherogenic Strain.

    Science.gov (United States)

    Filardo, Simone; Di Pietro, Marisa; Schiavoni, Giovanna; Minniti, Gianluca; Ortolani, Emanuela; Romano, Silvio; Sessa, Rosa

    2015-01-01

    Chlamydia pneumoniae has been associated to atherosclerotic cardiovascular diseases. The aim of our study was to characterize, for the first time, a C. pneumoniae strain isolated from the gingival crevicular fluid of a patient with chronic periodontitis, described as a risk factor for cardiovascular diseases. C. pneumoniae isolate was characterized and compared to the respiratory AR-39 strain by VD4-ompA genotyping and by investigating the intracellular growth in epithelial and macrophage cell lines and its ability to induce macrophage-derived foam cells. Inflammatory cytokine levels were determined in the gingival crevicular fluid sample. C. pneumoniae isolate showed a 99% similarity with the AR-39 strain in the VD4-ompA gene sequence and shared a comparable growth kinetic in epithelial cells and macrophages, as evidenced by the infectious progeny and by the number of chlamydial genomic copies. C. pneumoniae isolate significantly increased the number of foam cells as compared to uninfected and LDL-treated macrophages (45 vs. 6%, P = 0.0065) and to the AR-39 strain (45 vs. 30%, P = 0.0065). Significantly increased levels of interleukin 1-β (2.1 ± 0.3 pg/μL) and interleukin 6 (0.6 ± 0.08 pg/μL) were found. Our results suggest that C. pneumoniae may harbor inside oral cavity and potentially be atherogenic, even though further studies will be needed to clarify the involvement of C. pneumoniae in chronic periodontitis as a risk factor for cardiovascular diseases.

  5. Complete genome sequence of a virulent Streptococcus agalactiae strain 138P isolated from diseased Nile tilapia

    Science.gov (United States)

    Streptococcus agalactiae strain 138P was isolated from the kidney of diseased Nile tilapia in Idaho during a 2007 streptococcal disease outbreak. The full genome of S. agalactiae 138P is 1,838,716 bp. The availability of this genome will allow comparative genomics to identify genes for antigen disco...

  6. Identification and typing of the yeast strains isolated from bili bili, a ...

    African Journals Online (AJOL)

    Seventy six yeast strains isolated form bili bili and others sample were identified and typed in purpose of selecting appropriate starter culture. Identification techniques included conventional phenetic method, PCR/RFLP of NTS2 rDNA region, partial sequencing of the D1/D2 region of 26S rDNA and karyotyping using ...

  7. Isolation and characterization of Rhizobium sp. strain YS-1r that degrades lignin in plant biomass.

    Science.gov (United States)

    Jackson, C A; Couger, M B; Prabhakaran, M; Ramachandriya, K D; Canaan, P; Fathepure, B Z

    2017-04-01

    The aim of this work was to isolate novel lignin-degrading organisms. Several pure cultures of bacteria that degrade lignin were isolated from bacterial consortia developed from decaying biomass. Among the isolates, Rhizobium sp. strain YS-1r (closest relative of Rhizobium petrolearium strain SL-1) was explored for its lignin-degrading ability. Microcosm studies showed that strain YS-1r was able to degrade a variety of lignin monomers, dimers and also native lignin in switchgrass and alfalfa. The isolate demonstrated lignin peroxidase (LiP) activity when grown on alkali lignin, p-anisoin, switchgrass or alfalfa, and only negligible activity was measured in glucose-grown cells suggesting inducible nature of the LiP activity. Analysis of the strain YS-1r genome revealed the presence of a variety of genes that code for various lignin-oxidizing, H 2 O 2 -producing as well as polysaccharide-hydrolysing enzymes. This study shows both the genomic and physiological capability of bacteria in the genus Rhizobium to metabolize lignin and lignin-like compounds. This is the first detailed report on the lignocellulose-degrading ability of a Rhizobium species and thus this study expands the role of alpha-proteobacteria in the degradation of lignin. The organism's ability to degrade lignin is significant since Rhizobia are widespread in soil, water and plant rhizospheres and some fix atmospheric nitrogen and also have the ability to degrade aromatic hydrocarbons. © 2017 The Society for Applied Microbiology.

  8. Complete genome sequence of a Klebsiella pneumoniae strain isolated from a known cotton insect boll vector

    Science.gov (United States)

    Klebsiella pneumoniae (associated with bacterial pneumonia) was previously isolated from Nezara viridula, a significant vector of cotton boll-rot pathogens. We provide the first annotated genome sequence of the cotton opportunistic strain K. pneumoniae 5-1. This data provides guidance to study the...

  9. Induction, isolation, and characterization of aspergillus niger mutant strains producing elevated levels of beta-galactosidase.

    OpenAIRE

    Nevalainen, K M

    1981-01-01

    An Aspergillus niger mutant strain, VTT-D-80144, with an improvement of three- to fourfold in the production of extracellular beta-galactosidase was isolated after mutagenesis. The production of beta-galactosidase by this mutant was unaffected by fermentor size, and the enzyme was also suitable for immobilization.

  10. Induction, isolation, and characterization of aspergillus niger mutant strains producing elevated levels of beta-galactosidase.

    Science.gov (United States)

    Nevalainen, K M

    1981-01-01

    An Aspergillus niger mutant strain, VTT-D-80144, with an improvement of three- to fourfold in the production of extracellular beta-galactosidase was isolated after mutagenesis. The production of beta-galactosidase by this mutant was unaffected by fermentor size, and the enzyme was also suitable for immobilization. PMID:6784672

  11. Use of lactic strains isolated from Algerian ewe's milk in the ...

    African Journals Online (AJOL)

    SERVER

    2008-04-17

    Apr 17, 2008 ... Accepted 29 February, 2008. Fifty three strains of thermophilic and mesophilic lactic acid bacteria were isolated from the ewe's milk. ... at 10 and 45°C (cocci) or 15 and 45°C (rods), and gas production from MRS-broth (Falsen .... therefore certainly rates amino acids) in the medium. In addition, during the ...

  12. Isolation and identification of a cold-adapted lipase producing strain ...

    African Journals Online (AJOL)

    A cold-adapted lipase producing strain of mesophilic bacterium, named SYBC LIP-Y, was isolated from the decayed seeds of Ginkgo biloba L. by screening with plates containing Victoria Blue B and with the flask-shaking fermentation. It was identified as a novel Burkholderia species. The properties of its lipase after ...

  13. Isolation and identification of a cold-adapted lipase producing strain ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-05-03

    May 3, 2010 ... A cold-adapted lipase producing strain of mesophilic bacterium, named SYBC LIP-Y, was isolated from the decayed seeds of Ginkgo biloba L. by screening with plates containing Victoria Blue B and with the flask-shaking fermentation. It was identified as a novel Burkholderia species. The properties of its.

  14. The sensitivity to antibiotics of strains of group B streptococcus isolates from pregnant women in Belgrade

    Directory of Open Access Journals (Sweden)

    Jovanović Luka

    2016-01-01

    Full Text Available Introduction: Group B streptococcus (GBS is a significant human pathogen. GBS colonizes the vagina and it is one of the most important causes of early neonatal sepsis and meningitis. In many countries, screening of pregnant women and intrapartal use of antibiotics are common practice. Macrolide and lincosamide resistant strains of GBS are a significant problem, because these antibiotics are the second line therapy in case of penicillin allergy. Aim: Our aim was to investigate the frequency of antibiotic resistant strains of GBS and to detect macrolide resistance phenotypes in GBS strains obtained from pregnant women in Belgrade. Material and Methods: 105 GBS isolates were obtained from vaginal swabs of pregnant women attending two Gynecology and Obstetrics Centers in Belgrade. The isolates were tested for antimicrobial susceptibility pattern and D test were performed on Mueller Hinton agar. Results: Macrolide and lincosamide resistance was found in 30.4 %, and 23.8 % of isolates, respectively. There was a high frequency of tetracycline resistant strains (88.6 %. Most frequent macrolide resistant phenotype was iMLSb (macrolide and inducibile lincosamide resistance (62.4%. Conclusion: The results of our study indicate that there is a high level of macrolide resistance among GBS isolates in Serbia and the active surveillance is needed.

  15. Isolation of bacterial strains able to metabolize lignin and lignin-related compounds.

    Science.gov (United States)

    Tian, J-H; Pourcher, A-M; Peu, P

    2016-07-01

    In this study, we identified five strains isolated from soil and sediments able to degrade kraft lignin, aromatic dyes and lignin derivatives. Using 16S rRNA gene sequencing, the isolates were identified as Serratia sp. JHT01, Serratia liquefacien PT01, Pseudomonas chlororaphis PT02, Stenotrophomonas maltophilia PT03 and Mesorhizobium sp. PT04. All the isolates showed significant growth on lignin with no water-extractable compounds. Synthetic aromatic dyes were used to assess the presence of oxidative enzymes. All the isolates were able to use the thiazine dye Methylene blue and the anthraquinone dye Remazol Brilliant Blue R as the sole carbon source. Guaiacol, veratryl alcohol and biphenyl were also mineralized by all the strains isolated. These results suggest they could be used for the treatment of aromatic pollutants and for the degradation of the lignocellulosic biomass. The valorization of waste lignin and lignocellulosic biomass by biocatalysis opens up new possibilities for the production of value-added substituted aromatics, biofuel and for the treatment of aromatic pollutants. Bacteria with ligninolytic potential could be a source of novel enzymes for controlled lignin depolymerization. In this work, five soil bacteria were isolated and studied. Every isolate showed significant growth on lignin and was able to degrade several lignin monomers and ligninolytic indicator dyes. They could thus be a source of novel ligninolytic enzymes as well as candidates for a bacterial consortium for the delignification of lignocellulosic biomass. © 2016 The Society for Applied Microbiology.

  16. Isolation and characterization of Bacillus thuringiensis strains active against Elasmopalpus lignosellus (Zeller, 1848 (Lepidoptera, Pyralidae

    Directory of Open Access Journals (Sweden)

    Janaina Zorzetti

    2017-08-01

    Full Text Available Elasmopalpus lignosellus (Zeller, 1848 (Lepidoptera, Pyralidae is an insect pest of 60 economically important crops, including sugarcane, wheat, soybean, rice, beans, sorghum, peanuts, and cotton. The aim of this work was to select and characterize Bacillus thuringiensis isolates with insecticidal activity against E. Lignosellus that could be used as an alternative method of control. Selective bioassays were done to evaluate the toxicity of 47 isolates against first instar larvae of E. lignosellus. For the most toxic bacterial strains, the lethal concentration (LC50 was estimated and morphological, biochemical and molecular methods were used to characterize the isolates. Among the 47 isolates tested, 12 caused mortality above 85% and showed LC50 values from 0.038E+8 to 0.855E+8 spores mL-1. Isolates BR83, BR145, BR09, BR78, S1534, and S1302 had the lowest LC50 values and did not differ from the standard HD-1 strain; the exception was BR83.The protein profiles produced bands with molecular masses of 60-130 kDa. The genes cry1, cry2, cry3, and cry11 were identified in the molecular characterization. The morphological analysis identified three different crystal inclusions: bipyramidal, spherical and cuboidal. Among the tested isolates, 12 isolates have potential for biotechnological control of E. Lignosellus by development of new biopesticides or genetically modified plants.

  17. [Investigation of the virulence genes in methicillin-resistant Staphylococcus aureus strains isolated from biomaterial surfaces].

    Science.gov (United States)

    Sudağidan, Mert; Cavuşoğlu, Cengiz; Bacakoğlu, Feza

    2008-01-01

    Staphylococci are the most important agents of nosocomial infections originating from biomaterials. The aim of this study was to investigate the presence of virulence genes and their phenotypic expressions in 11 methicillin-resistant Staphylococcus aureus strains isolated from the surfaces of clinically used biomaterials of 48 thorasic intensive-care unit patients. By the use of specific primers, the presence of genes encoding the attachment and biofilm production (icaA, icaC, bap), methicillin resistance (mecA), enterotoxins A-E (sea, seb, sec, sed, see), toxic shock syndrome toxin (tst), exfoliative toxins A and B (eta and etb), alpha- and beta-hemolysins (hla and hlb), staphylococcal exotoxin-like protein-1 (set1), proteases (sspA, sspB, aur, serine proteaz gene), lipase (geh) and the regulatory genes (sarA and agrCA) were investigated by polymerase chain reaction (PCR). The phenotypic properties of the isolates such as biofilm formation, antibiotic susceptibility, extracellular protease and lipase production were also evaluated. None of the isolates were found to be biofilm and/or slime producers, however, all strains were found to have icaA gene which is responsible for biofilm formation. Nevertheless the presence of icaC and bap genes that are also responsible for biofilm formation were not detected. All the strains have had mecA gene and were resistant to oxacillin, penicilin G and gentamicin, while 10 were also resistant to erythromycin and nine were also resistant to ofloxacin. The isolates were susceptible to vancomycin, teicoplanin and co-trimoxazole. Screening of toxin and regulatory genes revealed that all the strains harboured sea, set1, hla, hlb and sarA genes. The phenotypic tests for the determination of extracellular protease production revealed that all the strains formed very weak zones on skim milk and milk agar plates, and yielded negative results on casein agar plates. Furthermore, all strains were found to harbour sspA, sspB, aur and serine

  18. Isolation and characterization of a high-efficiency erythromycin A-degrading Ochrobactrum sp. strain.

    Science.gov (United States)

    Zhang, Weiwei; Qiu, Lina; Gong, Aijun; Yuan, Xiaotao

    2017-01-30

    In this work, Erythromycin A(EA)- degrading bacteria was isolated from the contaminated soil obtained from a pharmaceutical factory in China. The isolate designated as strain WX-J1 was identified as Ochrobactrum sp. by sequence analysis of its 16S rDNA gene. It can grow in a medium containing EA as the sole source of carbon and its optimal growth pH and temperature were 6.5 and 32°C, respectively. Under these conditions, when the initial Erythromycin A concentration was 100mg·L-1, 97% of Erythromycin A has been degraded. HPLC-MS analyses indicated that Erythromycin A degradation produced intermediates contained the following three substances: 3-depyranosyloxy erythromycin A, 7,12-dyhydroxy-6-deoxyerythronolide B, 6-deoxyerythronolide B and propionaldehyde. Since Erythromycin A-degrading Ochrobactrum sp. strain rapidly degraded Erythromycin A, this strain might be useful for bioremediation purposes. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. IMMUNOLOGIC STUDY OF STRAINS OF BACILLUS PFEIFFERI ISOLATED FROM A CASE OF MENINGITIS.

    Science.gov (United States)

    Anderson, R A; Schultz, O T

    1921-04-30

    Five strains of Bacillus pfeifferi were isolated from a case of meningitis. These strains came from the spinal fluid, blood, nose, throat, and nasopharynx. Immunologic reactions show no definite relations between these strains, although those from the nose, throat, and nasopharynx might be presumed to be related to one another. It is also presumable that the spinal fluid strain was derived from the upper air passages, and that the blood was invaded from the meninges. In spite of immunologic differences, it is believed that the five strains were genetically related. The variations in these five strains from a single case are as great as those which have been found by others for strains from different cases, or individuals. As determined by immunologic reactions, the number of so called strains of Bacillus pfeifferi is apparently limited only by the number of cultures which have been or might be isolated. It is inconceivable that under the designation Bacillus pfeifferi is included a heterogeneous mixture of innumerable distinct races of bacteria; there must be some biological relation which cannot be established by agglutination, complement fixation, or opsonin reactions. The variations which have been noted, may be an indication of a degree of instability which may have some bearing upon questions relating to the epidemiology of Bacillus pfeifferi infections and to the serum therapy of influenzal meningitis. The strain variations which have been shown to exist by immunologic methods do not support the theory of the etiological relationn of Bacillus pfeifferi to epidemic influenza, unless it can be shown that such variations are due to instability of the organism. The fact that the incidence of meningeal involvement was only slightly increased during the last pandemic is also evidence against the causal relation of the organism to the pandemic disease.

  20. Molecular and virulence characterization of Toxoplasma gondii strains isolated from humans in Portugal.

    Science.gov (United States)

    Vilares, Anabela; Gargaté, Maria João; Ferreira, Idalina; Martins, Susana; Gomes, João Paulo

    2017-03-01

    Toxoplasma gondii is an apicomplexan parasite responsible for toxoplasmosis which infects all warm-blooded vertebrates, including mammals and birds. The majority of studies conducted in Europe have revealed that more than 80 % of strains isolated from human infections belong to genotype II, whereas genotypes I and III are responsible for a small number of cases. Atypical and recombinant strains are generally associated with more severe infections. In Portugal, there is a lack of data concerning genetic diversity as the classical typing studies in humans have never been performed. We aimed to determine the Sag2 and microsatellite-based (TUB2, TgM-A, W35, B17, B18) genotypes of T. gondii isolated from humans in Portugal, as well as to study their virulence in mice. We analyzed 48 strains from congenital and acquired toxoplasmosis collected during the last two decades. Sag2-based genotyping of T. gondii was achieved in all 48 strains where 35 (73 %) were classified as type II and 13 (27 %) were type I. The multilocus PCR of five microsatellites allowed the classification of 10 strains (21 %) as recombinant strains that had been previously identified as type II or I by Sag2 typing. Seven out of the 48 strains, including three type I, three recombinant, and one type I, were virulent in mice. This study constitutes the first evidence of recombinant strains circulating in Portugal in humans from congenital infection, highlighting the need for a better evaluation of these strains as their phenotype is still barely understood.

  1. Molecular and Phenotypic Characterization of Diarrheagenic Escherichia coli Strains Isolated from Bacteremic Children.

    Science.gov (United States)

    Riveros, Maribel; García, Wilfredo; García, Coralith; Durand, David; Mercado, Erik; Ruiz, Joaquim; Ochoa, Theresa J

    2017-11-01

    Escherichia coli is an important cause of Gram-negative bacteremia. The aim of this study was to characterize at the molecular and phenotypic levels E. coli strains belonging to different diarrheagenic pathotypes [diarrheagenic E. coli (DEC)] isolated from bacteremia in children younger than 5 years of age. Seventy bacteremia E. coli strains were collected in a prospective study in 12 hospitals in Lima, Peru. The presence of virulence genes associated with DEC [enterotoxigenic (lt and st), enteropathogenic (eaeA), shiga toxin-producing (stx1and stx2), enteroinvasive (ipaH), enteroaggregative (aggR), and diffusely adherent (daaD)] was determined by multiplex real-time polymerase chain reaction (PCR). Those positive E. coli strains were further analyzed for 18 additional virulence factors encoding genes and others phenotypic features. Virulence genes associated with DEC were identified in seven bacteremic children (10%), including: one aggR-positive [enteroaggregative E. coli (EAEC)], one eaeA-positive [enteropathogenic E. coli (EPEC)], one st-positive [enterotoxigenic E. coli (ETEC)], one daaD-positive [diffusely adherent E. coli (DAEC)], and three strain positive for aggR and daaD (EAEC/DAEC) at the same time. All strains, except EPEC, had the Ag43 adhesin, and all, except ETEC had the siderophore gene fyuA. The phylogenetic profile of these strains was variable, two (B2), two (D), two (A), and one (B1) strain. These isolates were susceptible to all tested antibacterial agents except to ampicillin and gentamicin. The three EAEC/DAEC strains showed biofilm formation and aggregative adhesion and had the same repetitive extragenic palindromic-PCR patterns. These findings suggest that some DEC strains, especially agg-R and daa-D positive, might cause bacteremia in children.

  2. Technological Aptitude and Applications of Leuconostoc mesenteroides Bioactive Strains Isolated from Algerian Raw Camel Milk

    Science.gov (United States)

    Benmechernene, Zineb; Chentouf, Hanane Fatma; Yahia, Bellil; Fatima, Ghazi; Quintela-Baluja, Marcos; Calo-Mata, Pilar; Barros-Velázquez, Jorge

    2013-01-01

    Two strains (B7 and Z8) of the Leuconostoc mesenteroides subspecies mesenteroides that were isolated from Algerian camel milk from an initial pool of 13 strains and demonstrated a high ability to inhibit the growth of Listeria spp. were selected and characterised at the phenotypic and genotypic levels. Probiotic profiling and inhibition spectra against food borne pathogens in mixed cultures were also investigated. The bacteriocin produced by L. mesenteroides strain B7 was identified as leucocin B by specific PCR. In vitro studies demonstrated that both Leuconostoc mesenteroides strains exhibited a marked probiotic profile, showing high survival at low pH (2-3 and 4) in the presence of 0.5%, 1%, and 2% of bile salts and at pH 3 in the presence of 3 mg/mL pepsin. Susceptibility testing against antimicrobial agents was also performed for both strains. When tested in a mixed culture with Listeria innocua, Listeria ivanovii, or Staphylococcus aureus, strain B7 reduced the numbers of these species by 1.87, 1.78, and 1.38 log units, respectively. Consequently, these two strains were found to possess good probiotic properties in vitro and a high capacity for Listeria spp. inhibition in mixed cultures. Therefore, these strains have a favourable technological aptitude and a potential application as novel probiotic starters. PMID:24392451

  3. [Isolation, identification and characterization of a microcystin-degrading bacterium Paucibacter sp. strain CH].

    Science.gov (United States)

    You, Di-Jie; Chen, Xiao-Guo; Xiang, Hui-Yi; Ouyang, Liao; Yang, Bing

    2014-01-01

    A bacterium capable of degrading microcystin (MC), strain CH, was isolated from the sediment of Lake Chaohu, China. Strain CH was tentatively identified as Paucibacter sp. based on the analysis of 16S rRNA gene sequences. Paucibacter sp. strain CH can use microcystin LR (MCLR) as the sole carbon and energy sources, and 11.6 microg x mL(-1) of MCLR was degraded to below the detection limit within 10 hours with the first-order reaction rate constant of 0.242 h(-1). The optimum temperature and initial pH for MC degradation were 25-30 degrees C and pH 6-9, respectively. A novel intermediate product containing the Adda residue was detected during the degradation of MCLR, which is different from those produced by strain ACM-3962, and Adda was recognized as the final product of the degradation process. Furthermore, no homologue to any of the four genes, mlrA, mlrB, mlrC and mlrD previously associated with the degradation of MCLR by strain ACM-3962 was found in strain CH. These findings suggest that Paucibacter sp. strain CH mighe degrade MC through a different pathway from that of strain ACM-3962.

  4. Technological aptitude and applications of Leuconostoc mesenteroides bioactive strains isolated from Algerian raw camel milk.

    Science.gov (United States)

    Benmechernene, Zineb; Chentouf, Hanane Fatma; Yahia, Bellil; Fatima, Ghazi; Quintela-Baluja, Marcos; Calo-Mata, Pilar; Barros-Velázquez, Jorge

    2013-01-01

    Two strains (B7 and Z8) of the Leuconostoc mesenteroides subspecies mesenteroides that were isolated from Algerian camel milk from an initial pool of 13 strains and demonstrated a high ability to inhibit the growth of Listeria spp. were selected and characterised at the phenotypic and genotypic levels. Probiotic profiling and inhibition spectra against food borne pathogens in mixed cultures were also investigated. The bacteriocin produced by L. mesenteroides strain B7 was identified as leucocin B by specific PCR. In vitro studies demonstrated that both Leuconostoc mesenteroides strains exhibited a marked probiotic profile, showing high survival at low pH (2-3 and 4) in the presence of 0.5%, 1%, and 2% of bile salts and at pH 3 in the presence of 3 mg/mL pepsin. Susceptibility testing against antimicrobial agents was also performed for both strains. When tested in a mixed culture with Listeria innocua, Listeria ivanovii, or Staphylococcus aureus, strain B7 reduced the numbers of these species by 1.87, 1.78, and 1.38 log units, respectively. Consequently, these two strains were found to possess good probiotic properties in vitro and a high capacity for Listeria spp. inhibition in mixed cultures. Therefore, these strains have a favourable technological aptitude and a potential application as novel probiotic starters.

  5. Technological Aptitude and Applications of Leuconostoc mesenteroides Bioactive Strains Isolated from Algerian Raw Camel Milk

    Directory of Open Access Journals (Sweden)

    Zineb Benmechernene

    2013-01-01

    Full Text Available Two strains (B7 and Z8 of the Leuconostoc mesenteroides subspecies mesenteroides that were isolated from Algerian camel milk from an initial pool of 13 strains and demonstrated a high ability to inhibit the growth of Listeria spp. were selected and characterised at the phenotypic and genotypic levels. Probiotic profiling and inhibition spectra against food borne pathogens in mixed cultures were also investigated. The bacteriocin produced by L. mesenteroides strain B7 was identified as leucocin B by specific PCR. In vitro studies demonstrated that both Leuconostoc mesenteroides strains exhibited a marked probiotic profile, showing high survival at low pH (2-3 and 4 in the presence of 0.5%, 1%, and 2% of bile salts and at pH 3 in the presence of 3 mg/mL pepsin. Susceptibility testing against antimicrobial agents was also performed for both strains. When tested in a mixed culture with Listeria innocua, Listeria ivanovii, or Staphylococcus aureus, strain B7 reduced the numbers of these species by 1.87, 1.78, and 1.38 log units, respectively. Consequently, these two strains were found to possess good probiotic properties in vitro and a high capacity for Listeria spp. inhibition in mixed cultures. Therefore, these strains have a favourable technological aptitude and a potential application as novel probiotic starters.

  6. Identification of mosquito larvicidal bacterial strains isolated from north Sinai in Egypt

    Science.gov (United States)

    2012-01-01

    In the present study, two of the most toxic bacterial strains of Bacillus sphaericus against mosquito were identified with the most recent genetic techniques. The PCR product profiles indicated the presence of genes encoding Bin A, Bin B and Mtx1 in all analyzed strains; they are consistent with protein profiles. The preliminary bioinformatics analysis of the binary toxin genes sequence revealed that the open reading frames had high similarities when matched with nucleotides sequence in the database of other B. sphaericus strains. The biological activity of B. sphaericus strains varied according to growing medium, and cultivation time. The highest yield of viable counts, spores and larvicidal protein were attained after 5 days. Poly (P) medium achieved the highest yield of growth, sporulation, protein and larvicidal activity for all tested strains compared to the other tested media. The larvicidal protein produced by local strains (B. sphaericus EMCC 1931 and EMCC 1932) in P medium was more lethal against the 3rd instar larvae of Culex pipiens than that of reference strains (B. sphaericus 1593 and B. sphaericus 2297). The obtained results revealed that P medium was the most effective medium and will be used in future work in order to optimize large scale production of biocide by the locally isolated Bacillus sphaericus strains. PMID:22280528

  7. Isolation and identification of Mycoplasma mycoides cluster strains from goats in Chongqing, China

    Directory of Open Access Journals (Sweden)

    Wang Haoju

    2014-03-01

    Full Text Available In order to evaluate the prevalence of the Mycoplasma mycoides cluster in goats in Chongqing, China, an epidemiological survey in this area was carried out. A total of 68 samples were subjected to bacteria isolation on Hartley’s medium. Four isolates (three from lung tissue and one from nasal discharges were recovered from the samples and identified as the Mycoplasma species by their morphological and biochemical characteristics. They were further confirmed by PCR using 16S rRNA specific primer pairs and by restriction enzyme analysis. In vitro antimicrobial susceptibility of the isolates indicated that some strains had developed resistance to the antibiotics tested. This is the first report on the isolation, identification, and molecular characterisation of Mycoplasma species isolated from goats in Chongqing. This study also revealed a prevalence of Mycoplasma species infection in goats in this area.

  8. Genomic differentiation among two strains of the PS1 clade isolated from geographically separated marine habitats

    KAUST Repository

    Jimenez Infante, Francy M.

    2014-05-22

    Using dilution-to-extinction cultivation, we isolated a strain affiliated with the PS1 clade from surface waters of the Red Sea. Strain RS24 represents the second isolate of this group of marine Alphaproteobacteria after IMCC14465 that was isolated from the East (Japan) Sea. The PS1 clade is a sister group to the OCS116 clade, together forming a putatively novel order closely related to Rhizobiales. While most genomic features and most of the genetic content are conserved between RS24 and IMCC14465, their average nucleotide identity (ANI) is < 81%, suggesting two distinct species of the PS1 clade. Next to encoding two different variants of proteorhodopsin genes, they also harbor several unique genomic islands that contain genes related to degradation of aromatic compounds in IMCC14465 and in polymer degradation in RS24, possibly reflecting the physicochemical differences in the environment they were isolated from. No clear differences in abundance of the genomic content of either strain could be found in fragment recruitment analyses using different metagenomic datasets, in which both genomes were detectable albeit as minor part of the communities. The comparative genomic analysis of both isolates of the PS1 clade and the fragment recruitment analysis provide first insights into the ecology of this group. © 2014 Federation of European Microbiological Societies.

  9. Molecular characterization of four Haemophilus influenzae serotype a strains isolated from patients in Quebec, Canada.

    Science.gov (United States)

    Sill, Michelle L; Zhou, Jianwei; Law, Dennis K S; Lorange, Manon; Ringuette, Louise; Bekal, Sadjia; Tsang, Raymond S W

    2007-10-01

    Four epidemiologically unrelated Haemophilus influenzae serotype a (Hia) strains from patients in Quebec, Canada, were characterized and found to represent 3 distinct groups. One isolate, found to be biotype I and sequence type (ST)-62 by multilocus sequence typing, was shown to possess the copper- and zinc-containing superoxide dismutase gene, sodC, and was suspected to belong to clonal division II. The other 3 isolates were classified as clonal division I based on the absence of the sodC gene. Among the 3 sodC-negative Hia strains, 2 were biotype II and had related STs (ST-23 and ST-403) and highly similar DNA fingerprints, similar to a group of previously described Hia isolates causing invasive disease in Manitoba, Canada. The remaining sodC-negative strain belonged to biotype I and ST-4 and shared no common allele with ST-23, ST-403, or ST-62. This isolate also possessed the IS1016-bexA partial deletion, which is often associated with increased virulence. Despite the small number of isolates used in this study, our finding of 3 distinct groups shows the existence of a potential genetic diversity not previously described for Hia. Whether this genetic diversity is related to the severity and epidemiology of Hia disease requires further studies.

  10. GROWTH CAPACITY OF BACILLUS POTENTIAL STARTER STRAINS ISOLATED FROM COCOA BEANS FERMENTATION UNDER CULTURE STRESS CONDITIONS

    Directory of Open Access Journals (Sweden)

    WILFRIED YAO

    2017-06-01

    Full Text Available The study investigates the growth capacity of Bacillus with pectinolytic activity, acidifying and citrate metabolism capacities under culture stress conditions. Spontaneous heaps fermentation was conducted from cocoa of six producing regions of Côte d’Ivoire. Bacillus isolated using plate agar on nutrient medium were analyzed for pectinolytic enzymes production, citric acid breakdown, acidification and growth capacity under different stress conditions. A total of 970 Bacillus strains were isolated and 44.53 % of them produced pectinolytic activity. Among pectinolytic strains, 163 (37.73 % exhibited acidifying and citrate metabolism capacity. Five (5 of these strains exhibited a strong thermotolerance at 50 ºC with the optimal growth at 35 ºC and a good capacity to grow at pH 4 to pH 8. Moreover, sugar concentrations ranged 5 to 25 % showed low effect on all tested strains growth with a maximum growth at 5 % fructose and sucrose concentration and at 15% glucose concentration. However, ethanol stress conditions (up to 8 % repress strongly growth capacity of the strains analyzed. This study indicates that Bacillus strains involved in Ivorian cocoa fermentation possess some properties essential for a well-fermented cocoa. Therefore, these results show that Bacillus studied should be potential candidate as starter for cocoa beans fermentation control.

  11. Comparative analysis of Pasteurella multocida strains isolated from bovine respiratory infections.

    Science.gov (United States)

    Sellyei, Boglárka; Rónai, Zsuzsanna; Jánosi, Szilárd; Makrai, László

    2015-12-01

    Bovine respiratory disease (BRD) is the leading cause of significant economic losses in the intensive beef industry worldwide. Beside numerous risk factors Pasteurella multocida, which is regarded as a secondary pathogen, may play a role in the development of the disease. Previous studies of strains from swine pneumonia revealed that there are a few clones associated with clinical disease, suggesting that some strains may be more virulent than others. This linkage may be true in the BRD, however composition of P. multocida populations in the herds are slightly characterized. Thus, we decided to perform phenotypic and genotypic characterisation of strains isolated from calves with respiratory infection at 31 different herds in Hungary. The results demonstrated the presence of two dominant strain types. At the identical taxonomic background (P. multocida subsp. multocida) with slight phenotypic variability they could be separated by trehalose fermentation capacity, α-glucosidase activity and molecular fingerprint patterns of ERIC- and M13-PCR. Independent prevalence and geographical origin of the strain types may refer to their significance in the illness, but their comparison with strains isolated from healthy individuals is taken into consideration.

  12. PCR detection of cytK gene in Bacillus cereus group strains isolated from food samples.

    Science.gov (United States)

    Oltuszak-Walczak, Elzbieta; Walczak, Piotr

    2013-11-01

    A method for detection of the cytotoxin K cytK structural gene and its active promoter preceded by the PlcR-binding box, controlling the expression level of this enterotoxin, was developed. The method was applied for the purpose of the analysis of 47 bacterial strains belonging to the Bacillus cereus group isolated from different food products. It was found that the majority of the analyzed strains carried the fully functional cytK gene with its PlcR regulated promoter. The cytK gene was not detected in four emetic strains of Bacillus cereus carrying the cesB gene and potentially producing an emetic toxin - cereulide. The cytotoxin K gene was detected in 4 isolates classified as Bacillus mycoides and one reference strain B. mycoides PCM 2024. The promoter region and the N-terminal part of the cytK gene from two strains of B. mycoides (5D and 19E) showed similarities to the corresponding sequences of Bacillus cereus W23 and Bacillus thuringiensis HD-789, respectively. It was shown for the first time that the cytK gene promoter region from strains 5D and 19E of Bacillus mycoides had a similar arrangement to the corresponding sequence of Bacillus cereus ATCC 14579. The presence of the cytK gene in Bacillus mycoides shows that this species, widely recognized as nonpathogenic, may pose potential biohazard to human beings. © 2013.

  13. Isolation of pathogenic Yersinia enterocolitica strains from different sources in Izmir region, Turkey.

    Science.gov (United States)

    Bozcal, Elif; Uzel, Atac; Aydemir, Sohret; Skurnik, Mikael

    2015-11-01

    Yersinia enterocolitica is a foodborne pathogen that is very rarely encountered in Turkey. In this work, several human, porcine, and environmental samples collected from Izmir region in Turkey were examined for the presence of Y. enterocolitica using different cultivation and enrichment methods. A total of nine pathogenic Y. enterocolitica strains were isolated; five strains from pig stool and manure samples and four strains from waste water samples. On the other hand, no Y. enterocolitica was isolated from human diarrheal stool samples (n = 102) and from 12 gulf, canal, municipal pool, and well water samples. Biochemical and serological characterization of the nine Y. enterocolitica strains revealed that they belonged to three different bioserotypes: 4/O:3, 2/O:9, and 2/O:5,27. All the strains were deemed pathogenic based on virulence factor-specific PCR analysis. Detection of pathogenic Y. enterocolitica strains from the pig and waste water samples from the Izmir region indicates that Y. enterocolitica is a potential risk for public health.

  14. [Isolation and characterization of a sodium dodecyl benzene sulfonate degrading bacterial strain].

    Science.gov (United States)

    Wu, Chu

    2006-12-01

    A bacterial strain, designated as WZR-A, which could utilize sodium dodecyl benzene sulfonate (SDBS) as sole carbon and energy source for growth, was isolated from contaminated river. The strain was identified as Ochrobactrum anthropi based on its morphological and physiological properties, and 16S rDNA sequence analysis. The optimum pH and temperature for cell growth and SDBS degradation were 7.0 and 30 degrees U, respectively. The degradation rate of SDBS by strain WZR-A was higher than 80% when its concentration was lower than 400mg/L. The results of whole cell protein SDS-PAGE electrophoresis showed that there were very obvious differences in the total cell protein composition of the strain between before and after SDBS induction. The enzyme distribution experiment showed that the enzymes relative SDBS degradation in the bacterium was intracellular one. Results from the characterization of degradation substrates together with the detection of activities of relative catabolic enzymes in crude extracts indicated that the aromatic ring cracking of SDBS by the strain probably via the modified ortho cleavage pathway and the strain could use broader spectrum substrates. A large plasmid was detected by utilizing plasmid isolating and curing technique and it was found that the genes involved in SDBS degradation were likely located on the plasmid.

  15. Polymorphism of viral dsRNA in Xanthophyllomyces dendrorhous strains isolated from different geographic areas

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    Libkind Diego

    2009-10-01

    Full Text Available Abstract Background Strains of the astaxanthin producing yeast Xanthophyllomyces dendrorhous have been isolated from different cold regions around the earth, and the presence of double stranded RNA (dsRNA elements was described in some isolates. This kind of viruses is widely distributed among yeasts and filamentous fungi and, although generally are cryptic in function, their studies have been a key factor in the knowledge of important fungi. In this work, the characterization and genetic relationships among dsRNA elements were determined in strains representatives of almost all regions of the earth where X. dendrorhous have been isolated. Results Almost all strains of X. dendrorhous analyzed carry one, two or four dsRNA elements, of molecular sizes in the range from 0.8 to 5.0 kb. Different dsRNA-patterns were observed in strains with different geographic origin, being L1 (5.0 kb the common dsRNA element. By hybridization assays a high genomic polymorphism was observed among L1 dsRNAs of different X. dendrorhous strains. Contrary, hybridization was observed between L1 and L2 dsRNAs of strains from same or different regions, while the dsRNA elements of minor sizes (M, S1, and S2 present in several strains did not show hybridization with neither L1 or L2 dsRNAs. Along the growth curve of UCD 67-385 (harboring four dsRNAs an increase of L2 relative to L1 dsRNA was observed, whiles the S1/L1 ratio remains constant, as well as the M/L1 ratio of Patagonian strain. Strains cured of S2 dsRNA were obtained by treatment with anisomycin, and comparison of its dsRNA contents with uncured strain, revealed an increase of L1 dsRNA while the L2 and S1 dsRNA remain unaltered. Conclusion The dsRNA elements of X. dendrorhous are highly variable in size and sequence, and the dsRNA pattern is specific to the geographic region of isolation. Each L1 and L2 dsRNA are viral elements able to self replicate and to coexist into a cell, and L1 and S2 dsRNAs elements could

  16. Polymorphism of viral dsRNA in Xanthophyllomyces dendrorhous strains isolated from different geographic areas.

    Science.gov (United States)

    Baeza, Marcelo; Sanhueza, Mario; Flores, Oriana; Oviedo, Vicente; Libkind, Diego; Cifuentes, Víctor

    2009-10-08

    Strains of the astaxanthin producing yeast Xanthophyllomyces dendrorhous have been isolated from different cold regions around the earth, and the presence of double stranded RNA (dsRNA) elements was described in some isolates. This kind of viruses is widely distributed among yeasts and filamentous fungi and, although generally are cryptic in function, their studies have been a key factor in the knowledge of important fungi. In this work, the characterization and genetic relationships among dsRNA elements were determined in strains representatives of almost all regions of the earth where X. dendrorhous have been isolated. Almost all strains of X. dendrorhous analyzed carry one, two or four dsRNA elements, of molecular sizes in the range from 0.8 to 5.0 kb. Different dsRNA-patterns were observed in strains with different geographic origin, being L1 (5.0 kb) the common dsRNA element. By hybridization assays a high genomic polymorphism was observed among L1 dsRNAs of different X. dendrorhous strains. Contrary, hybridization was observed between L1 and L2 dsRNAs of strains from same or different regions, while the dsRNA elements of minor sizes (M, S1, and S2) present in several strains did not show hybridization with neither L1 or L2 dsRNAs. Along the growth curve of UCD 67-385 (harboring four dsRNAs) an increase of L2 relative to L1 dsRNA was observed, while the S1/L1 ratio remains constant, as well as the M/L1 ratio of Patagonian strain. Strains cured of S2 dsRNA were obtained by treatment with anisomycin, and comparison of its dsRNA contents with uncured strain, revealed an increase of L1 dsRNA while the L2 and S1 dsRNA remain unaltered. The dsRNA elements of X. dendrorhous are highly variable in size and sequence, and the dsRNA pattern is specific to the geographic region of isolation. Each L1 and L2 dsRNA are viral elements able to self replicate and to coexist into a cell, and L1 and S2 dsRNAs elements could be part of a helper/satellite virus system in X

  17. Exploring the Genomic Traits of Non-toxigenic Vibrio parahaemolyticus Strains Isolated in Southern Chile

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    Daniel Castillo

    2018-02-01

    Full Text Available Vibrio parahaemolyticus is the leading cause of seafood-borne gastroenteritis worldwide. As reported in other countries, after the rise and fall of the pandemic strain in Chile, other post-pandemic strains have been associated with clinical cases, including strains lacking the major toxins TDH and TRH. Since the presence or absence of tdh and trh genes has been used for diagnostic purposes and as a proxy of the virulence of V. parahaemolyticus isolates, the understanding of virulence in V. parahaemolyticus strains lacking toxins is essential to detect these strains present in water and marine products to avoid possible food-borne infection. In this study, we characterized the genome of four environmental and two clinical non-toxigenic strains (tdh-, trh-, and T3SS2-. Using whole-genome sequencing, phylogenetic, and comparative genome analysis, we identified the core and pan-genome of V. parahaemolyticus of strains of southern Chile. The phylogenetic tree based on the core genome showed low genetic diversity but the analysis of the pan-genome revealed that all strains harbored genomic islands carrying diverse virulence and fitness factors or prophage-like elements that encode toxins like Zot and RTX. Interestingly, the three strains carrying Zot-like toxin have a different sequence, although the alignment showed some conserved areas with the zot sequence found in V. cholerae. In addition, we identified an unexpected diversity in the genetic architecture of the T3SS1 gene cluster and the presence of the T3SS2 gene cluster in a non-pandemic environmental strain. Our study sheds light on the diversity of V. parahaemolyticus strains from the southern Pacific which increases our current knowledge regarding the global diversity of this organism.

  18. Physiological and symbiotic diversity of Cupriavidus necator strains isolated from nodules of Leguminosae species

    Directory of Open Access Journals (Sweden)

    Ligiane Aparecida Florentino

    2012-01-01

    Full Text Available Biological nitrogen fixation, performed by diazotrophic bacteria, plays an important role in the maintenance of agricultural systems, as it contributes with significant amounts of the nitrogen (N needed for plant growth, totally or partially exempting the use of industrial N fertilisers. Twenty-five bacterial strains isolated from nodules of Leucaena leucocephala and Phaseolus vulgaris trap plants were studied. These nodules were formed after inoculation with suspensions of soil samples collected close to the root system of Sesbania virgata. In previous studies, these bacteria were identified as Cupriavidus necator. This study aimed to evaluate the ability of these strains to fix N2 in the free-living state and to use carbon (C sources; their resistance to antibiotics; growth in media with different pH values and salt concentrations and symbiotic efficiency with L. leucocephala and P. vulgaris. In each test, these strains were compared to C. taiwanensis LMG 19424T. Although a high variability regarding antibiotic resistance, salt tolerance and use of C sources were observed among the 25 C. necator strains, a large group behaved similar regarding salt tolerance (20 strains and antibiotic resistance (22 strains. C. necator strains behaved in a different way of LMG 19424T. Only one of the 25 strains studied, UFLA02-69, was not able to establish symbiosis with its trap species, P. vulgaris. Only the strains LMG19424T and UFLA01-672 were efficient in symbiosis with L. leucocephala. The ability to use C sources, grow in different pHs and salt concentrations and resistance to several antibiotics, may grant high saprophytic competence and greater competitivity to these strains in relation to the native Leguminosae-nodulating bacterial populations, suggesting potential use in inoculant strain selection studies for legumes cultivated in soils with a wide range of pH and salt concentrations.

  19. [Antimicrobial sensitivity of Escherichia coli strains with K1 antigen isolated from pregnant women and newborns].

    Science.gov (United States)

    Kaczmarek, Agnieszka; Budzyńska, Anna; Gospodarek, Eugenia

    2011-01-01

    The aim of this study was comparison of the susceptibility to antibiotics of E. coli strains with K1 antigen (E. coli K1+) and non-K1 E. coli strains (E. coli K1-). This study included 67 of E. coli K1+ and 67 of E. coli K1- strains isolated in the time period from June to September of 2008 from pregnant women and newborns hospitalized at dr. J. Biziel University Hospital number 2 L. Rydygier Collegium Medicum in Bydgoszcz Nicolaus Copernicus University in Toruń. Antimicrobial susceptibility of E. coli strains was tested by the disc-diffusion method, on the Mueller Hinton 2 Agar (Becton Dickinson). It was found that 64,2% of E. coli K1+ strains and 53,7% of E. coli K1-strains were susceptible to all tested antibiotics and chemioterapeutics. E. coli K1- strains were more often than E. coli K1+ nonsusceptible to at least one antimicrobial agent. The obtained results indicate that E. coli K1+ strains significant differed in the susceptibility to ampicillin/sulbactam (85,1% versus 95,5%) (p=0,041), cephalothin (70,1% versus 85,1%) (p=0,038) and tetracycline (91,0% versus 74,6%) (p=0,012) from E. coli K1-strains. All tested E. coli K1+ and K1-strains were sensitive to piperacillin/tazobactam, cefoperazone/sulbactam, cefotaxime, ceftazidime, cefepime, imipenem, amikacin, netilmicin and tigecycline. There weren't the ESBL-producing strains among tested E. coli K1+ and K1- rods.

  20. Characterization of Trypanosoma rangeli Strains Isolated in Central and South America: an Overview

    Directory of Open Access Journals (Sweden)

    Grisard Edmundo C

    1999-01-01

    Full Text Available Trypanosoma rangeli is a hemoflagelate parasite that infects domestic and sylvatic animals, as well as man, in Central and South America. T. rangeli has an overlapping distribution with T. cruzi, the etiological agent of Chagas disease, sharing several animal reservoirs and triatomine vectors. We have isolated T. rangeli strains in the State of Santa Catarina, in southern Brazil, which dramatically increased the distribution area of this parasite. This brief review summarizes several studies comparing T. rangeli strains isolated in Santa Catarina with others isolated in Colombia, Honduras and Venezuela. The different methods used include indirect immunofluorescence and western blot assays, lectin agglutination, isoenzyme electrophoresis and random amplified polymorphic DNA analysis, triatomine susceptibility, in vitro cell infection assays, and mini-exon gene analysis.

  1. Isolation and Characterization of a Bacteriocin-Like Substance Produced by Geobacillus toebii Strain HBB-247.

    Science.gov (United States)

    Başbülbül Özdemir, Gamze; Biyik, Haci Halil

    2012-03-01

    A total of 201 thermophilic bacteria isolated from various thermal spring, mud and soil were tested for their antibacterial activity. Among the mostly active isolates, Geobacillus toebii HBB-247 was further examined. Bacteriocin-like inhibitory substance (BLIS) produced by strain HBB-247 was found to be stable up to 60°C, sensitive to proteolytic enzymes and effective against Enterococcus faecalis, Listeria sp., E. avium, Clostridium pasteurianum, Cellulomonas fimi and some thermophilic strains isolated and identified in this study. As a result of Tricine-SDS-PAGE molecular weight of BLIS was estimated about 38 kDa. Production studies showed that G. toebii HBB-247 starts to produce antibacterial substance at early logarithmic phase of growth and maximum production was detected at the end of the logarithmic phase.

  2. Characterization of Thermotolerant Chitinases Encoded by a Brevibacillus laterosporus Strain Isolated from a Suburban Wetland

    Directory of Open Access Journals (Sweden)

    Pulin Liu

    2015-12-01

    Full Text Available To isolate and characterize chitinases that can be applied with practical advantages, 57 isolates of chitin-degrading bacteria were isolated from the soil of a suburban wetland. 16S rRNA gene analysis revealed that the majority of these strains belonged to two genera, Paenibacillus and Brevibacillus. Taking thermostability into account, the chitinases (ChiA and ChiC of a B. laterosporus strain were studied further. Ni-NTA affinity-purified ChiA and ChiC were optimally active at pH 7.0 and 6.0, respectively, and showed high temperature stability up to 55 °C. Kinetic analysis revealed that ChiC has a lower affinity and stronger catalytic activity toward colloidal chitin than ChiA. With their stability in a broad temperature range, ChiA and ChiC can be utilized for the industrial bioconversion of chitin wastes into biologically active products.

  3. [Genomic characteristics of coxsackievirus B5 A210/KM/09 strain isolated in Yunnan, China].

    Science.gov (United States)

    Liu, Jiansheng; Shao, Congwen; Zhao, Weizhong; Zhang, Yunkun; Ji, Ma; Zhu, Yanju; Ma, Zhongfei; Ma, Shaohui

    2014-03-01

    To characterize the complete genome sequence of coxsackievirus B5 (CVB5)A210/KM/09 strain which was isolated from Yunnan, China, 2009. Eight overlapping clones covering the whole viral genome (excluding the poly-A tail)were obtained by RT-PCR and sequenced, with their nucleotide and amino acid sequences compared with other known CVB5 strains. The genome of the CVB5 A210/KM/09 strain had 7 372 nucleotides in length, and containing a 742-nt non-translated region (NTR) at the 5' end and a 98-nt NTR at the 3' end. The entire open reading frame contained 6 555 nt, encoding a 2 185-aa polyprotein. In the coding region, there appeared no nucleotide deletion or insertion, but some changes of amino acid seemed unique. Based on the complete genome sequence alignments, CVB5 isolate A210/KM/09 strain showed the highest nucleotide (92.5%) and amino acid (97.3%) identities to the CVB5/CC10/10. It also shared nucleotide (80.1%-92.5%) and amino acid (95.0%-97.3%) homology with other CVB5 strains: 17Y, 19CSF, 20CSF, 1954/85/US, 2000/CSF/KOR, 03001N, CoxB5/Henan/2010, VB5/SD/09 and Faulkner. Blast between genome fragments, A210/KM/09 showed similarity on nucleotide (80.1%-92.5%) and amino acid (95.0%-97.3%) identities with other CVB5 strains. The phylogenetic tree, constructed on the complete VP1 regions, indicated that CVB5 could be divided into genotype A, B, C and D. while Genotype C and D could be further divided into C1-C4 and D1-D4 subgenotypes. A210/KM/09 and other CVB5 predominant strains isolated in China belonged to CVB5 subgenotype C4.

  4. Genetic diversity of environmental Vibrio cholerae O1 strains isolated in Northern Vietnam.

    Science.gov (United States)

    Takemura, Taichiro; Murase, Kazunori; Maruyama, Fumito; Tran, Thi Luong; Ota, Atsushi; Nakagawa, Ichiro; Nguyen, Dong Tu; Ngo, Tu Cuong; Nguyen, Thi Hang; Tokizawa, Asako; Morita, Masatomo; Ohnishi, Makoto; Nguyen, Binh Minh; Yamashiro, Tetsu

    2017-10-01

    Cholera epidemics have been recorded periodically in Vietnam during the seventh cholera pandemic. Since cholera is a water-borne disease, systematic monitoring of environmental waters for Vibrio cholerae presence is important for predicting and preventing cholera epidemics. We conducted monitoring, isolation, and genetic characterization of V. cholerae strains in Nam Dinh province of Northern Vietnam from Jul 2013 to Feb 2015. In this study, four V. cholerae O1 strains were detected and isolated from 110 analyzed water samples (3.6%); however, none of them carried the cholera toxin gene, ctxA, in their genomes. Whole genome sequencing and phylogenetic analysis revealed that the four O1 isolates were separated into two independent clusters, and one of them diverged from a common ancestor with pandemic strains. The analysis of pathogenicity islands (CTX prophage, VPI-I, VPI-II, VSP-I, and VSP-II) indicated that one strain (VNND_2014Jun_6SS) harbored an unknown prophage-like sequence with high homology to vibriophage KSF-1 phi and VCY phi, identified from Bangladesh and the USA, respectively, while the other three strains carried tcpA gene with a distinct sequence demonstrating a separate clonal lineage. These results suggest that the aquatic environment can harbor highly divergent V. cholera strains and serve as a reservoir for multiple V. cholerae virulence-associated genes which may be exchanged via mobile genetic elements. Therefore, continuous monitoring and genetic characterization of V. cholerae strains in the environment should contribute to the early detection of the sources of infection and prevention of cholera outbreaks as well as to understanding the natural ecology and evolution of V. cholerae. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  5. Induction of antigen-specific Th1-type immune responses by gamma-irradiated recombinant Brucella abortus RB51.

    Science.gov (United States)

    Sanakkayala, Neelima; Sokolovska, Anna; Gulani, Jatinder; Hogenesch, Harm; Sriranganathan, Nammalwar; Boyle, Stephen M; Schurig, Gerhardt G; Vemulapalli, Ramesh

    2005-12-01

    Brucella abortus strain RB51 is an attenuated rough mutant used as the live vaccine against bovine brucellosis in the United States and other countries. We previously reported the development of strain RB51 as a bacterial vaccine vector for inducing Th1-type immune responses against heterologous proteins. Because safety concerns may preclude the use of strain RB51-based recombinant live vaccines, we explored the ability of a gamma-irradiated recombinant RB51 strain to induce heterologous antigen-specific immune responses in BALB/c mice. Exposure of strain RB51G/LacZ expressing Escherichia coli beta-galactosidase to a minimum of 300 kilorads of gamma radiation resulted in complete loss of replicative ability. These bacteria, however, remained metabolically active and continued to synthesize beta-galactosidase. A single intraperitoneal inoculation of mice with 10(9) CFU equivalents of gamma-irradiated, but not heat-killed, RB51G/LacZ induced a beta-galactosidase-specific Th1-type immune response. Though no obvious differences were detected in immune responses to B. abortus-specific antigens, mice vaccinated with gamma-irradiated, but not heat-killed, RB51G/LacZ developed significant protection against challenge with virulent B. abortus. In vitro experiments indicated that gamma-irradiated and heat-killed RB51G/LacZ induced maturation of dendritic cells; however, stimulation with gamma-irradiated bacteria resulted in more interleukin-12 secretion. These results suggest that recombinant RB51 strains exposed to an appropriate minimum dose of gamma radiation are unable to replicate but retain their ability to stimulate Th1 immune responses against the heterologous antigens and confer protection against B. abortus challenge in mice.

  6. Brucella abortus agglutinins in dogs in Zaria, Nigeria | Osinubi ...

    African Journals Online (AJOL)

    Serum samples from dogs brought for routine physical examination, vaccination and other complaints at the Veterinary Teaching Hospital (VTH) of Ahmadu Bello University and other parts of Zaria were tested for Brucella abortus antibodies. Forty-three (21.5%) of the 200 dogs studied were positive for B. abortus agglutinins ...

  7. Characterization of trh2 Harbouring Vibrio parahaemolyticus Strains Isolated in Germany

    Science.gov (United States)

    Bechlars, Silke; Jäckel, Claudia; Diescher, Susanne; Wüstenhagen, Doreen A.; Kubick, Stefan; Dieckmann, Ralf; Strauch, Eckhard

    2015-01-01

    Background Vibrio parahaemolyticus is a recognized human enteropathogen. Thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) as well as the type III secretion system 2 (T3SS2) are considered as major virulence factors. As tdh positive strains are not detected in coastal waters of Germany, we focused on the characterization of trh positive strains, which were isolated from mussels, seawater and patients in Germany. Results Ten trh harbouring V. parahaemolyticus strains from Germany were compared to twenty-one trh positive strains from other countries. The complete trh sequences revealed clustering into three different types: trh1 and trh2 genes and a pseudogene Ψtrh. All German isolates possessed alleles of the trh2 gene. MLST analysis indicated a close relationship to Norwegian isolates suggesting that these strains belong to the autochthonous microflora of Northern Europe seawaters. Strains carrying the pseudogene Ψtrh were negative for T3SS2β effector vopC. Transcription of trh and vopC genes was analyzed under different growth conditions. Trh2 gene expression was not altered by bile while trh1 genes were inducible. VopC could be induced by urea in trh2 bearing strains. Most trh1 carrying strains were hemolytic against sheep erythrocytes while all trh2 positive strains did not show any