WorldWideScience

Sample records for botulinum strains isolated

  1. An atypical Clostridium strain related to the Clostridium botulinum group III strain isolated from a human blood culture.

    Science.gov (United States)

    Bouvet, Philippe; Ruimy, Raymond; Bouchier, Christiane; Faucher, Nathalie; Mazuet, Christelle; Popoff, Michel R

    2014-01-01

    A nontoxigenic strain isolated from a fatal human case of bacterial sepsis was identified as a Clostridium strain from Clostridium botulinum group III, based on the phenotypic characters and 16S rRNA gene sequence, and was found to be related to the mosaic C. botulinum D/C strain according to a multilocus sequence analysis of 5 housekeeping genes.

  2. 一株C型肉毒梭菌的分离和鉴别%The isolation and identification of a type C strain of clostridium botulinum

    Institute of Scientific and Technical Information of China (English)

    冉枭; 钟权涛; 傅思武

    2013-01-01

    OBJECTIVE The study is examined for distribution of clostridium botulinum in Northwest,China.METHODS All soils from Gansu Province were washed by 0.2% gelatin phosphate buffer,and then carried out with the methods of bacteria-proliferating,isolating culture repeatedly.The isolated strain was identified by toxin determination and biochemical characteristics.RESULTS 2 of the 175 soil specimens appeared typical symptoms of botulism,and only 1 could be completely neutralized by type C botulinum antitoxin.CONCLUSION The strain is identified as type C clostridium botulinum.%目的 为了解肉毒梭菌在我国西北地区的分布情况.方法 对从甘肃省采集的所有土壤样品用0.2%的明胶磷酸盐缓冲液冲洗后,反复增菌、分离培养,并对分离菌株进行毒素测定和生化特性分析.结果 在采集的175份样品中有2份样品出现典型的肉毒中毒症状,1株与C型肉毒抗毒素完全中和.结论 分离得到的菌株经鉴定为C型肉毒梭菌.

  3. Genetic Diversity Among Botulinum Neurotoxin-Producing Clostridial Strains

    Science.gov (United States)

    2007-02-01

    fragment length polymorphism analysis of Norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl. Environ. Microbiol. 67:4863–4873...isolation and purification. Individual bacterial colonies of each of the C. botulinum strains were removed from anaerobic CDC blood agar plates and

  4. Molecular diversity of Clostridium botulinum and phenotypically similar strains.

    Science.gov (United States)

    Grenda, T; Kukier, E; Sieradzki, Z; Goldsztejn, M; Kwiatek, K

    2016-12-01

    This study was undertaken to examine phenotypic and genetic features of strains preliminary classified as Clostridium botulinum species. The phenotypic characteristics were assessed with different culture media and biochemical tests. The genetic characterization included detection of botulinum toxin genes by PCR and macrorestriction analysis with SmaI, XhoI and SacII by PFGE (Pulsed-field Gel Electrophoresis). Despite similar biochemical properties of all analysed strains, only 47% of them contained genes determining toxicity specific to C. botulinum species. The most valuable differentiation of C. botulinum and C. botulinum-like strains was obtained after SmaI digestion. The highest affinity was observed among C. botulinum type B profiles which was even up to 100%. It was found 100% of affinity between C. botulinum and C. botulinum-like strains, however, the similarity among C. botulinum and C. botulinum-like was generally lower than 80%.

  5. Identification and genetic characterization of Clostridium botulinum serotype A strains from commercially pasteurized carrot juice.

    Science.gov (United States)

    Marshall, Kristin M; Nowaczyk, Louis; Raphael, Brian H; Skinner, Guy E; Rukma Reddy, N

    2014-12-01

    Clostridium botulinum is an important foodborne pathogen capable of forming heat resistant endospores and producing deadly botulinum neurotoxins (BoNTs). In 2006, C. botulinum was responsible for an international outbreak of botulism attributed to the consumption of commercially pasteurized carrot juice. The purpose of this study was to isolate and characterize strains of C. botulinum from the adulterated product. Carrot juice bottles retrieved from the manufacturing facility were analyzed for the presence of BoNT and BoNT-producing isolates using DIG-ELISA. Toxigenic isolates from the carrot juice were analyzed using pulsed-field gel electrophoresis (PFGE) and DNA microarray analysis to determine their genetic relatedness to the original outbreak strains CDC51348 and CDC51303. PFGE revealed that isolates CJ4-1 and CJ10-1 shared an identical pulsotype with strain CDC51303, whereas isolate CJ5-1 displayed a unique restriction banding pattern. DNA microarray analysis identified several phage related genes unique to strain CJ5-1, and Southern hybridization analysis of XhoI digested and nondigested DNA showed their chromosomal location, while a homolog to pCLI_A009 of plasmid pCLI of C. botulinum serotype Langeland F, was located on a small plasmid. The acquisition or loss of bacteriophages and other mobile genetic elements among C. botulinum strains has epidemiological and evolutionary implications.

  6. Diversity of Group I and II Clostridium botulinum Strains from France Including Recently Identified Subtypes.

    Science.gov (United States)

    Mazuet, Christelle; Legeay, Christine; Sautereau, Jean; Ma, Laurence; Bouchier, Christiane; Bouvet, Philippe; Popoff, Michel R

    2016-06-13

    In France, human botulism is mainly food-borne intoxication, whereas infant botulism is rare. A total of 99 group I and II Clostridium botulinum strains including 59 type A (12 historical isolates [1947-1961], 43 from France [1986-2013], 3 from other countries, and 1 collection strain), 31 type B (3 historical, 23 recent isolates, 4 from other countries, and 1 collection strain), and 9 type E (5 historical, 3 isolates, and 1 collection strain) were investigated by botulinum locus gene sequencing and multilocus sequence typing analysis. Historical C. botulinum A strains mainly belonged to subtype A1 and sequence type (ST) 1, whereas recent strains exhibited a wide genetic diversity: subtype A1 in orfX or ha locus, A1(B), A1(F), A2, A2b2, A5(B2') A5(B3'), as well as the recently identified A7 and A8 subtypes, and were distributed into 25 STs. Clostridium botulinum A1(B) was the most frequent subtype from food-borne botulism and food. Group I C. botulinum type B in France were mainly subtype B2 (14 out of 20 historical and recent strains) and were divided into 19 STs. Food-borne botulism resulting from ham consumption during the recent period was due to group II C. botulinum B4. Type E botulism is rare in France, 5 historical and 1 recent strains were subtype E3. A subtype E12 was recently identified from an unusual ham contamination. Clostridium botulinum strains from human botulism in France showed a wide genetic diversity and seems to result not from a single evolutionary lineage but from multiple and independent genetic rearrangements.

  7. Evidence that plasmid-borne botulinum neurotoxin type B genes are widespread among Clostridium botulinum serotype B strains.

    Directory of Open Access Journals (Sweden)

    Giovanna Franciosa

    Full Text Available BACKGROUND: Plasmids that encode certain subtypes of the botulinum neurotoxin type B have recently been detected in some Clostridium botulinum strains. The objective of the present study was to investigate the frequency with which plasmid carriage of the botulinum neurotoxin type B gene (bont/B occurs in strains of C. botulinum type B, Ab, and A(B, and whether plasmid carriage is bont/B subtype-related. METHODOLOGY/PRINCIPAL FINDINGS: PCR-Restriction fragment length polymorphism was employed to identify subtypes of the bont/B gene. Pulsed-field gel electrophoresis and Southern blot hybridization with specific probes were performed to analyze the genomic location of the bont/B subtype genes. All five known bont/B subtype genes were detected among the strains; the most frequently detected subtype genes were bont/B1 and /B2. Surprisingly, the bont/B subtype gene was shown to be plasmid-borne in >50% of the total strains. The same bont/B subtype gene was associated with the chromosome in some strains, whereas it was associated with a plasmid in others. All five known bont/B subtype genes were in some cases found to reside on plasmids, though with varying frequency (e.g., most of the bont/B1 subtype genes were located on plasmids, whereas all but one of the bont/B2 subtypes were chromosomally-located. Three bivalent isolates carried both bont/A and /B genes on the same plasmid. The plasmids carrying the bont gene were five different sizes, ranging from approximately 55 kb to approximately 245 kb. CONCLUSIONS/SIGNIFICANCE: The unexpected finding of the widespread distribution of plasmids harboring the bont/B gene among C. botulinum serotype B strains provides a chance to examine their contribution to the dissemination of the bont genes among heterogeneous clostridia, with potential implications on issues related to pathogenesis and food safety.

  8. Genomics of Clostridium botulinum group III strains.

    Science.gov (United States)

    Sakaguchi, Yoshihiko; Suzuki, Tomonori; Yamamoto, Yumiko; Nishikawa, Atsushi; Oguma, Keiji

    2015-05-01

    In Clostridium botulinum, the characteristics of type C and D strains are quite different from other types, and they are classified as group III. They produce C2 binary toxin and C3 exoenzyme in addition to type C and D neurotoxins. Two different phages and many plasmids are identified in the organisms. The genes of neurotoxin and C3 exoenzyme are converted from toxigenic strains to non-toxigenic strains by the specific bacteriophages (phages), whereas, the C2 toxin gene is carried by large or small plasmids. Classification of type C and D strains has been in confusion because 1) antigenicity of type C and D neurotoxins is complex, 2) the cells produce two types of toxins, neurotoxin and C2 toxin, and 3) some non-toxigenic strains can be converted to produce C or D neurotoxin by the infection with phages. Until now, entire nucleotide sequences of cell chromosomes, phages, and plasmids have been determined. Since both genetic and protein-chemical analyses have been clarifying the above confusions, these data are reviewed historically.

  9. Flagellin Diversity in Clostridium botulinum Groups I and II: a New Strategy for Strain Identification▿

    Science.gov (United States)

    Paul, Catherine J.; Twine, Susan M.; Tam, Kevin J.; Mullen, James A.; Kelly, John F.; Austin, John W.; Logan, Susan M.

    2007-01-01

    Strains of Clostridium botulinum are traditionally identified by botulinum neurotoxin type; however, identification of an additional target for typing would improve differentiation. Isolation of flagellar filaments and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that C. botulinum produced multiple flagellin proteins. Nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) analysis of in-gel tryptic digests identified peptides in all flagellin bands that matched two homologous tandem flagellin genes identified in the C. botulinum Hall A genome. Designated flaA1 and flaA2, these open reading frames encode the major structural flagellins of C. botulinum. Colony PCR and sequencing of flaA1/A2 variable regions classified 80 environmental and clinical strains into group I or group II and clustered isolates into 12 flagellar types. Flagellar type was distinct from neurotoxin type, and epidemiologically related isolates clustered together. Sequencing a larger PCR product, obtained during amplification of flaA1/A2 from type E strain Bennett identified a second flagellin gene, flaB. LC-MS analysis confirmed that flaB encoded a large type E-specific flagellin protein, and the predicted molecular mass for FlaB matched that observed by SDS-PAGE. In contrast, the molecular mass of FlaA was 2 to 12 kDa larger than the mass predicted by the flaA1/A2 sequence of a given strain, suggesting that FlaA is posttranslationally modified. While identification of FlaB, and the observation by SDS-PAGE of different masses of the FlaA proteins, showed the flagellin proteins of C. botulinum to be diverse, the presence of the flaA1/A2 gene in all strains examined facilitates single locus sequence typing of C. botulinum using the flagellin variable region. PMID:17351097

  10. Systemic colonization of clover (Trifolium repens by Clostridium botulinum strain 2301

    Directory of Open Access Journals (Sweden)

    Matthias eZeiller

    2015-10-01

    Full Text Available In recent years, cases of botulism in cattle and other farm animals and also in farmers increased dramatically. It was proposed, that these cases could be affiliated with the spreading of compost or other organic manures contaminated with Clostridium botulinum spores on farm land. Thus, soils and fodder plants and finally farm animals could be contaminated. Therefore, the colonization behavior and interaction of the botulinum neurotoxin (BoNT D producing C. botulinum strain 2301 and the non-toxin producing Clostridium sporogenes strain 1739 were investigated on clover (Trifolium repens in a field experiment as well as in phytochamber experiments applying axenic and additionally soil based systems under controlled conditions. Plants were harvested and divided into root and shoot parts for further DNA isolation and PCR assays; subsamples were fixed for fluorescence in situ hybridization (FISH analysis in combination with confocal laser scanning microscopy (CLSM. To target C. botulinum and C. sporogenes, 16S rDNA directed primers were used and to specifically detect C. botulinum, BoNT D toxin genes targeted primers, using a multiplex PCR approach, were applied. Our results demonstrate an effective colonization of roots and shoots of clover by C. botulinum strain 2301 and C. sporogenes strain 1739. Detailed analysis of colonization behavior showed that C. botulinum can occur as individual cells, in cell clusters and in microcolonies within the rhizosphere, lateral roots and within the roots tissue of clover. In addition, we observed significant differences in the growth behavior of clover plants when inoculated with Clostridia spores, indicating a plant growth promoting effect. Inoculated plants showed an increased growth index (shoot size, wet and dry weight and an enlarged root system, which suggests the involvement of phytohormonal effects induced by the systemic colonization of clover by C. botulinum strain 2301.

  11. The same clade of Clostridium botulinum strains is causing avian botulism in southern and northern Europe.

    Science.gov (United States)

    Anza, Ibone; Skarin, Hanna; Vidal, Dolors; Lindberg, Anna; Båverud, Viveca; Mateo, Rafael

    2014-04-01

    Avian botulism is a paralytic disease caused by Clostridium botulinum-produced botulinum neurotoxins (BoNTs), most commonly of type C/D. It is a serious disease of waterbirds and poultry flocks in many countries in Europe. The objective of this study was to compare the genetic relatedness of avian C. botulinum strains isolated in Spain with strains isolated in Sweden using pulsed-field gel electrophoresis (PFGE). Fifteen strains were isolated from Spanish waterbirds using an immunomagnetic separation technique. Isolates were characterized by PCR, and all were identified as the genospecies Clostridium novyi sensu lato and eight harboured the gene coding for the BoNT type C/D. PFGE analysis of the strains revealed four highly similar pulsotypes, out of which two contained strains from both countries. It also showed that outbreaks in wild and domestic birds can be caused by the same strains. These results support a clonal spreading of the mosaic C. botulinum type C/D through Europe and give relevant information for future epidemiological studies.

  12. Genomic characterization of Italian Clostridium botulinum group I strains.

    Science.gov (United States)

    Giordani, Francesco; Fillo, Silvia; Anselmo, Anna; Palozzi, Anna Maria; Fortunato, Antonella; Gentile, Bernardina; Azarnia Tehran, Domenico; Ciammaruconi, Andrea; Spagnolo, Ferdinando; Pittiglio, Valentina; Anniballi, Fabrizio; Auricchio, Bruna; De Medici, Dario; Lista, Florigio

    2015-12-01

    Clostridium botulinum is a gram-positive bacterium capable of producing the botulinum neurotoxin, a powerful poison that causes botulism, a severe neuroparalytic disease. Its genome has been sequenced entirely and its gene content has been analyzed. To date, 19 full genomes and 64 draft genomes are available. The geographical origin of these genomes is predominantly from the US. In the present study, 10 Italian genomes of C. botulinum group I were analyzed and compared with previously sequenced group I genomes, in order to genetically characterize the Italian population of C. botulinum group I and to investigate the phylogenetic relationships among different lineages. Using the suites of software ClonalFrame and ClonalOrigin to perform genomic analysis, we demonstrated that Italian C. botulinum group I population is phylogenetically heterogeneous encompassing different and distant lineages including overseas strains, too. Moreover, a high recombination rate was demonstrated in the evolution of C. botulinum group I species. Finally, genome sequencing of the strain 357 led us to identify a novel botulinum neurotoxin subtype, F8.

  13. A penicillin- and metronidazole-resistant Clostridium botulinum strain responsible for an infant botulism case.

    Science.gov (United States)

    Mazuet, C; Yoon, E-J; Boyer, S; Pignier, S; Blanc, T; Doehring, I; Meziane-Cherif, D; Dumant-Forest, C; Sautereau, J; Legeay, C; Bouvet, P; Bouchier, C; Quijano-Roy, S; Pestel-Caron, M; Courvalin, P; Popoff, M R

    2016-07-01

    The clinical course of a case of infant botulism was characterized by several relapses despite therapy with amoxicillin and metronidazole. Botulism was confirmed by identification of botulinum toxin and Clostridium botulinum in stools. A C. botulinum A2 strain resistant to penicillins and with heterogeneous resistance to metronidazole was isolated from stool samples up to 110 days after onset. Antibiotic susceptibility was tested by disc agar diffusion and MICs were determined by Etest. Whole genome sequencing allowed detection of a gene cluster composed of blaCBP for a novel penicillinase, blaI for a regulator, and blaR1 for a membrane-bound penicillin receptor in the chromosome of the C. botulinum isolate. The purified recombinant penicillinase was assayed. Resistance to β-lactams was in agreement with the kinetic parameters of the enzyme. In addition, the β-lactamase gene cluster was found in three C. botulinum genomes in databanks and in two of 62 genomes of our collection, all the strains belonging to group I C. botulinum. This is the first report of a C. botulinum isolate resistant to penicillins. This stresses the importance of antibiotic susceptibility testing for adequate therapy of botulism.

  14. Growth of group II Clostridium botulinum strains at extreme temperatures.

    Science.gov (United States)

    Derman, Yağmur; Lindström, Miia; Selby, Katja; Korkeala, Hannu

    2011-11-01

    The minimum and maximum growth temperatures and the maximum growth rates at 10, 30, 37, and 40°C were determined for 24 group II Clostridium botulinum strains. Genetic diversity of the strains was revealed by amplified fragment length polymorphism (AFLP) analysis. The minimum growth temperatures ranged from 6.2 to 8.6°C, and the maximum growth temperatures ranged from 34.7 to 39.9°C. The mean maximum growth temperatures and mean maximum growth rates of type E strains at 37°C were significantly higher than those of type B and type F strains. A significant correlation between maximum growth rates at 37°C and maximum growth temperatures was found for all strains. Some type E strains with a high minimum growth temperature also had a higher maximum growth rate at 37°C than at 30°C, which suggests that some group II C. botulinum strains are more mesophilic in their growth properties than others. We found relatively small differences between AFLP clusters, indicating that diverse genetic background among the strains was not reflected in the growth properties. The growth characteristics of group II C. botulinum and some type E strains with mesophilic growth properties may have an impact on inoculation studies and predictive modeling for assessing the safety of foods.

  15. Clostridium botulinum Group II Isolate Phylogenomic Profiling Using Whole-Genome Sequence Data.

    Science.gov (United States)

    Weedmark, K A; Mabon, P; Hayden, K L; Lambert, D; Van Domselaar, G; Austin, J W; Corbett, C R

    2015-09-01

    Clostridium botulinum group II isolates (n = 163) from different geographic regions, outbreaks, and neurotoxin types and subtypes were characterized in silico using whole-genome sequence data. Two clusters representing a variety of botulinum neurotoxin (BoNT) types and subtypes were identified by multilocus sequence typing (MLST) and core single nucleotide polymorphism (SNP) analysis. While one cluster included BoNT/B4/F6/E9 and nontoxigenic members, the other comprised a wide variety of different BoNT/E subtype isolates and a nontoxigenic strain. In silico MLST and core SNP methods were consistent in terms of clade-level isolate classification; however, core SNP analysis showed higher resolution capability. Furthermore, core SNP analysis correctly distinguished isolates by outbreak and location. This study illustrated the utility of next-generation sequence-based typing approaches for isolate characterization and source attribution and identified discrete SNP loci and MLST alleles for isolate comparison.

  16. Clostridium botulinum Group I Strain Genotyping by 15-Locus Multilocus Variable-Number Tandem-Repeat Analysis ▿ †

    Science.gov (United States)

    Fillo, Silvia; Giordani, Francesco; Anniballi, Fabrizio; Gorgé, Olivier; Ramisse, Vincent; Vergnaud, Gilles; Riehm, Julia M.; Scholz, Holger C.; Splettstoesser, Wolf D.; Kieboom, Jasper; Olsen, Jaran-Strand; Fenicia, Lucia; Lista, Florigio

    2011-01-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of C. botulinum isolates of any BoNT type is relevant for both epidemiological studies and forensic microbiology. A 10-locus multiple-locus variable-number tandem-repeat analysis (MLVA) was previously applied to isolates of C. botulinum type A. The present study includes five additional loci designed to better address proteolytic B and F serotypes. We investigated 79 C. botulinum group I strains isolated from human and food samples in several European countries, including types A (28), B (36), AB (4), and F (11) strains, and 5 nontoxic Clostridium sporogenes. Additional data were deduced from in silico analysis of 10 available fully sequenced genomes. This 15-locus MLVA (MLVA-15) scheme identified 86 distinct genotypes that clustered consistently with the results of amplified fragment length polymorphism (AFLP) and MLVA genotyping in previous reports. An MLVA-7 scheme, a subset of the MLVA-15, performed on a lab-on-a-chip device using a nonfluorescent subset of primers, is also proposed as a first-line assay. The phylogenetic grouping obtained with the MLVA-7 does not differ significantly from that generated by the MLVA-15. To our knowledge, this report is the first to analyze genetic variability among all of the C. botulinum group I serotypes by MLVA. Our data provide new insights into the genetic variability of group I C. botulinum isolates worldwide and demonstrate that this group is genetically highly diverse. PMID:22012011

  17. Clostridium botulinum group I strain genotyping by 15-locus multilocus variable-number tandem-repeat analysis.

    Science.gov (United States)

    Fillo, Silvia; Giordani, Francesco; Anniballi, Fabrizio; Gorgé, Olivier; Ramisse, Vincent; Vergnaud, Gilles; Riehm, Julia M; Scholz, Holger C; Splettstoesser, Wolf D; Kieboom, Jasper; Olsen, Jaran-Strand; Fenicia, Lucia; Lista, Florigio

    2011-12-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of C. botulinum isolates of any BoNT type is relevant for both epidemiological studies and forensic microbiology. A 10-locus multiple-locus variable-number tandem-repeat analysis (MLVA) was previously applied to isolates of C. botulinum type A. The present study includes five additional loci designed to better address proteolytic B and F serotypes. We investigated 79 C. botulinum group I strains isolated from human and food samples in several European countries, including types A (28), B (36), AB (4), and F (11) strains, and 5 nontoxic Clostridium sporogenes. Additional data were deduced from in silico analysis of 10 available fully sequenced genomes. This 15-locus MLVA (MLVA-15) scheme identified 86 distinct genotypes that clustered consistently with the results of amplified fragment length polymorphism (AFLP) and MLVA genotyping in previous reports. An MLVA-7 scheme, a subset of the MLVA-15, performed on a lab-on-a-chip device using a nonfluorescent subset of primers, is also proposed as a first-line assay. The phylogenetic grouping obtained with the MLVA-7 does not differ significantly from that generated by the MLVA-15. To our knowledge, this report is the first to analyze genetic variability among all of the C. botulinum group I serotypes by MLVA. Our data provide new insights into the genetic variability of group I C. botulinum isolates worldwide and demonstrate that this group is genetically highly diverse.

  18. Whole-genome single-nucleotide-polymorphism analysis for discrimination of Clostridium botulinum group I strains.

    Science.gov (United States)

    Gonzalez-Escalona, Narjol; Timme, Ruth; Raphael, Brian H; Zink, Donald; Sharma, Shashi K

    2014-04-01

    Clostridium botulinum is a genetically diverse Gram-positive bacterium producing extremely potent neurotoxins (botulinum neurotoxins A through G [BoNT/A-G]). The complete genome sequences of three strains harboring only the BoNT/A1 nucleotide sequence are publicly available. Although these strains contain a toxin cluster (HA(+) OrfX(-)) associated with hemagglutinin genes, little is known about the genomes of subtype A1 strains (termed HA(-) OrfX(+)) that lack hemagglutinin genes in the toxin gene cluster. We sequenced the genomes of three BoNT/A1-producing C. botulinum strains: two strains with the HA(+) OrfX(-) cluster (69A and 32A) and one strain with the HA(-) OrfX(+) cluster (CDC297). Whole-genome phylogenic single-nucleotide-polymorphism (SNP) analysis of these strains along with other publicly available C. botulinum group I strains revealed five distinct lineages. Strains 69A and 32A clustered with the C. botulinum type A1 Hall group, and strain CDC297 clustered with the C. botulinum type Ba4 strain 657. This study reports the use of whole-genome SNP sequence analysis for discrimination of C. botulinum group I strains and demonstrates the utility of this analysis in quickly differentiating C. botulinum strains harboring identical toxin gene subtypes. This analysis further supports previous work showing that strains CDC297 and 657 likely evolved from a common ancestor and independently acquired separate BoNT/A1 toxin gene clusters at distinct genomic locations.

  19. Genetic Diversity Among Botulinum Neurotoxin Producing Clostridial Strains

    Energy Technology Data Exchange (ETDEWEB)

    Hill, K K; Smith, T J; Helma, C H; Ticknor, L O; Foley, B T; Svennson, R T; Brown, J L; Johnson, E A; Smith, L A; Okinaka, R T; Jackson, P J; Marks, J D

    2006-07-06

    Clostridium botulinum is a taxonomic designation for many diverse anaerobic spore forming rod-shaped bacteria which have the common property of producing botulinum neurotoxins (BoNTs). The BoNTs are exoneurotoxins that can cause severe paralysis and even death in humans and various other animal species. A collection of 174 C. botulinum strains were examined by amplified fragment length polymorphism (AFLP) analysis and by sequencing of the 16S rRNA gene and BoNT genes to examine genetic diversity within this species. This collection contained representatives of each of the seven different serotypes of botulinum neurotoxins (BoNT A-G). Analysis of the16S rRNA sequences confirmed earlier reports of at least four distinct genomic backgrounds (Groups I-IV) each of which has independently acquired one or more BoNT serotypes through horizontal gene transfer. AFLP analysis provided higher resolution, and can be used to further subdivide the four groups into sub-groups. Sequencing of the BoNT genes from serotypes A, B and E in multiple strains confirmed significant sequence variation within each serotype. Four distinct lineages within each of the BoNT A and B serotypes, and five distinct lineages of serotype E strains were identified. The nucleotide sequences of the seven serotypes of BoNT were compared and show varying degrees of interrelatedness and recombination as has been previously noted for the NTNH gene which is linked to BoNT. These analyses contribute to the understanding of the evolution and phylogeny within this species and assist in the development of improved diagnostics and therapeutics for treatment of botulism.

  20. Genetic diversity among Clostridium botulinum strains harboring bont/A2 and bont/A3 genes.

    Science.gov (United States)

    Lúquez, Carolina; Raphael, Brian H; Joseph, Lavin A; Meno, Sarah R; Fernández, Rafael A; Maslanka, Susan E

    2012-12-01

    Clostridium botulinum type A strains are known to be genetically diverse and widespread throughout the world. Genetic diversity studies have focused mainly on strains harboring one type A botulinum toxin gene, bont/A1, although all reported bont/A gene variants have been associated with botulism cases. Our study provides insight into the genetic diversity of C. botulinum type A strains, which contain bont/A2 (n = 42) and bont/A3 (n = 4) genes, isolated from diverse samples and geographic origins. Genetic diversity was assessed by using bont nucleotide sequencing, content analysis of the bont gene clusters, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). Sequences of bont genes obtained in this study showed 99.9 to 100% identity with other bont/A2 or bont/A3 gene sequences available in public databases. The neurotoxin gene clusters of the subtype A2 and A3 strains analyzed in this study were similar in gene content. C. botulinum strains harboring bont/A2 and bont/A3 genes were divided into six and two MLST profiles, respectively. Four groups of strains shared a similarity of at least 95% by PFGE; the largest group included 21 out of 46 strains. The strains analyzed in this study showed relatively limited genetic diversity using either MLST or PFGE.

  1. Immunoprecipitation of native botulinum neurotoxin complexes from Clostridium botulinum subtype A strains.

    Science.gov (United States)

    Lin, Guangyun; Tepp, William H; Bradshaw, Marite; Fredrick, Chase M; Johnson, Eric A

    2015-01-01

    Botulinum neurotoxins (BoNTs) naturally exist as components of protein complexes containing nontoxic proteins. The nontoxic proteins impart stability of BoNTs in the gastrointestinal tract and during purification and handling. The two primary neurotoxin complexes (TCs) are (i) TC1, consisting of BoNT, nontoxin-nonhemagglutinin (NTNH), and hemagglutinins (HAs), and (ii) TC2, consisting of BoNT and NTNH (and possibly OrfX proteins). In this study, BoNT/A subtypes A1, A2, A3, and A5 were examined for the compositions of their TCs in culture extracts using immunoprecipitation (IP). IP analyses showed that BoNT/A1 and BoNT/A5 form TC1s, while BoNT/A2 and BoNT/A3 form TC2s. A Clostridium botulinum host strain expressing recombinant BoNT/A4 (normally present as a TC2) from an extrachromosomal plasmid formed a TC1 with complexing proteins from the host strain, indicating that the HAs and NTNH encoded on the chromosome associated with the plasmid-encoded BoNT/A4. Strain NCTC 2916 (A1/silent B1), which carries both an ha silent bont/b cluster and an orfX bont/a1 cluster, was also examined. IP analysis revealed that NCTC 2916 formed only a TC2 containing BoNT/A1 and its associated NTNH. No association between BoNT/A1 and the nontoxic proteins from the silent bont/b cluster was detected, although the HAs were expressed as determined by Western blotting analysis. Additionally, NTNH and HAs from the silent bont/b cluster did not form a complex in NCTC 2916. The stabilities of the two types of TC differed at various pHs and with addition of KCl and NaCl. TC1 complexes were more stable than TC2 complexes. Mouse serum stabilized TC2, while TC1 was unaffected.

  2. Two-component systems are involved in the regulation of botulinum neurotoxin synthesis in Clostridium botulinum type A strain Hall.

    Science.gov (United States)

    Connan, Chloé; Brüggemann, Holger; Brueggemann, Holger; Mazuet, Christelle; Raffestin, Stéphanie; Cayet, Nadège; Popoff, Michel R

    2012-01-01

    Clostridium botulinum synthesizes a potent neurotoxin (BoNT) which associates with non-toxic proteins (ANTPs) to form complexes of various sizes. The bont and antp genes are clustered in two operons. In C. botulinum type A, bont/A and antp genes are expressed during the end of the exponential growth phase and the beginning of the stationary phase under the control of an alternative sigma factor encoded by botR/A, which is located between the two operons. In the genome of C. botulinum type A strain Hall, 30 gene pairs predicted to encode two-component systems (TCSs) and 9 orphan regulatory genes have been identified. Therefore, 34 Hall isogenic antisense strains on predicted regulatory genes (29 TCSs and 5 orphan regulatory genes) have been obtained by a mRNA antisense procedure. Two TCS isogenic antisense strains showed more rapid growth kinetics and reduced BoNT/A production than the control strain, as well as increased bacterial lysis and impairment of the bacterial cell wall structure. Three other TCS isogenic antisense strains induced a low level of BoNT/A and ANTP production. Interestingly, reduced expression of bont/A and antp genes was shown to be independent of botR/A. These results indicate that BoNT/A synthesis is under the control of a complex network of regulation including directly at least three TCSs.

  3. Two-component systems are involved in the regulation of botulinum neurotoxin synthesis in Clostridium botulinum type A strain Hall.

    Directory of Open Access Journals (Sweden)

    Chloé Connan

    Full Text Available Clostridium botulinum synthesizes a potent neurotoxin (BoNT which associates with non-toxic proteins (ANTPs to form complexes of various sizes. The bont and antp genes are clustered in two operons. In C. botulinum type A, bont/A and antp genes are expressed during the end of the exponential growth phase and the beginning of the stationary phase under the control of an alternative sigma factor encoded by botR/A, which is located between the two operons. In the genome of C. botulinum type A strain Hall, 30 gene pairs predicted to encode two-component systems (TCSs and 9 orphan regulatory genes have been identified. Therefore, 34 Hall isogenic antisense strains on predicted regulatory genes (29 TCSs and 5 orphan regulatory genes have been obtained by a mRNA antisense procedure. Two TCS isogenic antisense strains showed more rapid growth kinetics and reduced BoNT/A production than the control strain, as well as increased bacterial lysis and impairment of the bacterial cell wall structure. Three other TCS isogenic antisense strains induced a low level of BoNT/A and ANTP production. Interestingly, reduced expression of bont/A and antp genes was shown to be independent of botR/A. These results indicate that BoNT/A synthesis is under the control of a complex network of regulation including directly at least three TCSs.

  4. Genetic characterization and comparison of Clostridium botulinum isolates from botulism cases in Japan between 2006 and 2011.

    Science.gov (United States)

    Kenri, Tsuyoshi; Sekizuka, Tsuyoshi; Yamamoto, Akihiko; Iwaki, Masaaki; Komiya, Takako; Hatakeyama, Takashi; Nakajima, Hiroshi; Takahashi, Motohide; Kuroda, Makoto; Shibayama, Keigo

    2014-11-01

    Genetic characterization was performed for 10 group I Clostridium botulinum strains isolated from botulism cases in Japan between 2006 and 2011. Of these, 1 was type A, 2 were type B, and 7 were type A(B) {carrying a silent bont/B [bont/(B)] gene} serotype strains, based on botulinum neurotoxin (BoNT) production. The type A strain harbored the subtype A1 BoNT gene (bont/A1), which is associated with the ha gene cluster. The type B strains carried bont/B5 or bont/B6 subtype genes. The type A(B) strains carried bont/A1 identical to that of type A(B) strain NCTC2916. However, bont/(B) genes in these strains showed single-nucleotide polymorphisms (SNPs) among strains. SNPs at 2 nucleotide positions of bont/(B) enabled classification of the type A(B) strains into 3 groups. Pulsed-field gel electrophoresis (PFGE) and multiple-locus variable-number tandem-repeat analysis (MLVA) also provided consistent separation results. In addition, the type A(B) strains were separated into 2 lineages based on their plasmid profiles. One lineage carried a small plasmid (5.9 kb), and another harbored 21-kb plasmids. To obtain more detailed genetic information about the 10 strains, we sequenced their genomes and compared them with 13 group I C. botulinum genomes in a database using whole-genome SNP analysis. This analysis provided high-resolution strain discrimination and enabled us to generate a refined phylogenetic tree that provides effective traceability of botulism cases, as well as bioterrorism materials. In the phylogenetic tree, the subtype B6 strains, Okayama2011 and Osaka05, were distantly separated from the other strains, indicating genomic divergence of subtype B6 strains among group I strains.

  5. Molecular composition and extinction coefficient of native botulinum neurotoxin complex produced by Clostridium botulinum hall A strain.

    Science.gov (United States)

    Bryant, Anne-Marie; Davis, Jenny; Cai, Shuowei; Singh, Bal Ram

    2013-02-01

    Seven distinct strains of Clostridium botulinum (type A to G) each produce a stable complex of botulinum neurotoxin (BoNT) along with neurotoxin-associated proteins (NAPs). Type A botulinum neurotoxin (BoNT/A) is produced with a group of NAPs and is commercially available for the treatment of numerous neuromuscular disorders and cosmetic purposes. Previous studies have indicated that BoNT/A complex composition is specific to the strain, the method of growth and the method of purification; consequently, any variation in composition of NAPs could have significant implications to the effectiveness of BoNT based therapeutics. In this study, a standard analytical technique using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and densitometry analysis was developed to accurately analyze BoNT/A complex from C. botulinum type A Hall strain. Using 3 batches of BoNT/A complex the molar ratio was determined as neurotoxin binding protein (NBP, 124 kDa), heavy chain (HC, 90 kDa), light chain (LC, 53 kDa), NAP-53 (50 kDa), NAP-33 (36 kDa), NAP-22 (24 kDa), NAP-17 (17 kDa) 1:1:1:2:3:2:2. With Bradford, Lowry, bicinchoninic acid (BCA) and spectroscopic protein estimation methods, the extinction coefficient of BoNT/A complex was determined as 1.54 ± 0.26 (mg/mL)(-1)cm(-1). These findings of a reproducible BoNT/A complex composition will aid in understanding the molecular structure and function of BoNT/A and NAPs.

  6. Neurotoxins from Clostridium botulinum (serotype A) isolated from the soil of Mendoza (Argentina) differ from the A-Hall archetype and from that causing infant botulism.

    Science.gov (United States)

    Caballero, P; Troncoso, M; Patterson, S I; López Gómez, C; Fernandez, R; Sosa, M A

    2016-10-01

    The type A of neurotoxin produced by Clostridium botulinum is the prevalent serotype in strains of Mendoza. The soil is the main reservoir for C.botulinum and is possibly one of the infection sources in infant botulism. In this study, we characterized and compared autochthonous C. botulinum strains and their neurotoxins. Bacterial samples were obtained from the soil and from fecal samples collected from children with infant botulism. We first observed differences in the appearance of the colonies between strains from each source and with the A Hall control strain. In addition, purified neurotoxins of both strains were found to be enriched in a band of 300 kDa, whereas the A-Hall strain was mainly made up of a band of ∼600 kDa. This finding is in line with the lack of hemagglutinating activity of the neurotoxins under study. Moreover, the proteolytic activity of C. botulinum neurotoxins was evaluated against SNARE (soluble N-ethylmaleimide-sensitive factor-attachment protein receptor) proteins from rat brain. It was observed that both, SNAP 25 (synaptosomal-associated protein 25) and VAMP 2 (vesicle-associated membrane protein) were cleaved by the neurotoxins isolated from the soil strains, whereas the neurotoxins from infant botulism strains only induced a partial cleavage of VAMP 2. On the other hand, the neurotoxin from the A-Hall strain was able to cleave both proteins, though at a lesser extent. Our data indicate that the C.botulinum strain isolated from the soil, and its BoNT, exhibit different properties compared to the strain obtained from infant botulism patients, and from the A-Hall archetype.

  7. Effects of Megaplasmid Loss on Growth of Neurotoxigenic Clostridium butyricum Strains and Botulinum Neurotoxin Type E Expression

    OpenAIRE

    Concetta eScalfaro; Angelo eIacobino; Laura eGrande; Stefano eMorabito; Giovanna eFranciosa

    2016-01-01

    Clostridium butyricum strains that atypically produce the botulinum neurotoxin type E (BoNT/E) possess a megaplasmid of unknown functions in their genome. In this study, we cured two botulinum neurotoxigenic C. butyricum type E strains of their megaplasmids, and compared the obtained megaplasmid-cured strains to their respective wild-type parental strains. Our results showed that the megaplasmids do not confer beta-lactam resistance on the neurotoxigenic C. butyricum type E strains, although ...

  8. Biochemical classification of Clostridium botulinum type C and D strains and their nontoxigenic derivatives.

    OpenAIRE

    OGUMA, K.; Yamaguchi, T.; Sudou, K; Yokosawa, N; Fujikawa, Y.

    1986-01-01

    The biochemical properties of 11 toxigenic and 10 nontoxigenic type C and D strains of Clostridium botulinum were studied. All of the strains examined were motile and hemolytic and produced lipase and liquid gelatin. Fermentation of several sugars and the production of lecithinase, indole, and hydrogen sulfide varied with the strain. The strains were classified into four groups based on their sugar fermentation profiles. The resulting classification was identical to the classification which h...

  9. Recombination and Insertion Events Involving the Botulinum Neurotoxin Complex Genes in Clostridium botulinum Types A, B, E and F and Clostridium butyricum Type E Strains

    Science.gov (United States)

    2009-10-05

    Bruggemann H, Baumer S, Fricke WF, Wiezer A, Liesegang H, Decker I, et al.: The genome sequence of Clostridium tetani , the caus- ative agent of... Clostridium botulinum types A, B, E and F and Clostridium butyricum type E strains Karen K Hill*1, Gary Xie2, Brian T Foley3, Theresa J Smith4, Amy C Munk2...ornl.gov; John C Detter - cdetter@lanl.gov * Corresponding author Abstract Background: Clostridium botulinum is a taxonomic designation for at least

  10. Conjugative botulinum neurotoxin-encoding plasmids in Clostridium botulinum.

    Directory of Open Access Journals (Sweden)

    Kristin M Marshall

    Full Text Available BACKGROUND: Clostridium botulinum produces seven distinct serotypes of botulinum neurotoxins (BoNTs. The genes encoding different subtype neurotoxins of serotypes A, B, F and several dual neurotoxin-producing strains have been shown to reside on plasmids, suggesting that intra- and interspecies transfer of BoNT-encoding plasmids may occur. The objective of the present study was to determine whether these C. botulinum BoNT-encoding plasmids are conjugative. METHODOLOGY/PRINCIPAL FINDINGS: C. botulinum BoNT-encoding plasmids pBotCDC-A3 (strain CDC-A3, pCLJ (strain 657Ba and pCLL (strain Eklund 17B were tagged with the erythromycin resistance marker (Erm using the ClosTron mutagenesis system by inserting a group II intron into the neurotoxin genes carried on these plasmids. Transfer of the tagged plasmids from the donor strains CDC-A3, 657Ba and Eklund 17B to tetracycline-resistant recipient C. botulinum strains was evaluated in mating experiments. Erythromycin and tetracycline resistant transconjugants were isolated from donor:recipient mating pairs tested. Transfer of the plasmids to the transconjugants was confirmed by pulsed-field gel electrophoresis (PFGE and Southern hybridizations. Transfer required cell-to-cell contact and was DNase resistant. This indicates that transfer of these plasmids occurs via a conjugation mechanism. CONCLUSIONS/SIGNIFICANCE: This is the first evidence supporting conjugal transfer of native botulinum neurotoxin-encoding plasmids in C. botulinum, and provides a probable mechanism for the lateral distribution of BoNT-encoding plasmids to other C. botulinum strains. The potential transfer of C. botulinum BoNT-encoding plasmids to other bacterial hosts in the environment or within the human intestine is of great concern for human pathogenicity and necessitates further characterization of these plasmids.

  11. Genomic Epidemiology of Clostridium botulinum Isolates from Temporally Related Cases of Infant Botulism in New South Wales, Australia.

    Science.gov (United States)

    McCallum, Nadine; Gray, Timothy J; Wang, Qinning; Ng, Jimmy; Hicks, Leanne; Nguyen, Trang; Yuen, Marion; Hill-Cawthorne, Grant A; Sintchenko, Vitali

    2015-09-01

    Infant botulism is a potentially life-threatening paralytic disease that can be associated with prolonged morbidity if not rapidly diagnosed and treated. Four infants were diagnosed and treated for infant botulism in NSW, Australia, between May 2011 and August 2013. Despite the temporal relationship between the cases, there was no close geographical clustering or other epidemiological links. Clostridium botulinum isolates, three of which produced botulism neurotoxin serotype A (BoNT/A) and one BoNT serotype B (BoNT/B), were characterized using whole-genome sequencing (WGS). In silico multilocus sequence typing (MLST) found that two of the BoNT/A-producing isolates shared an identical novel sequence type, ST84. The other two isolates were single-locus variants of this sequence type (ST85 and ST86). All BoNT/A-producing isolates contained the same chromosomally integrated BoNT/A2 neurotoxin gene cluster. The BoNT/B-producing isolate carried a single plasmid-borne bont/B gene cluster, encoding BoNT subtype B6. Single nucleotide polymorphism (SNP)-based typing results corresponded well with MLST; however, the extra resolution provided by the whole-genome SNP comparisons showed that the isolates differed from each other by >3,500 SNPs. WGS analyses indicated that the four infant botulism cases were caused by genomically distinct strains of C. botulinum that were unlikely to have originated from a common environmental source. The isolates did, however, cluster together, compared with international isolates, suggesting that C. botulinum from environmental reservoirs throughout NSW have descended from a common ancestor. Analyses showed that the high resolution of WGS provided important phylogenetic information that would not be captured by standard seven-loci MLST.

  12. First report of an infant botulism case due to Clostridium botulinum type Af.

    Science.gov (United States)

    de Jong, Laura I T; Fernández, Rafael A; Pareja, Virtudes; Giaroli, Gabriel; Guidarelli, Sergio R; Dykes, Janet K; Lúquez, Carolina

    2015-02-01

    Most infant botulism cases worldwide are due to botulinum toxin types A and B. Rarely, Clostridium botulinum strains that produce two serotypes (Ab, Ba, and Bf) have also been isolated from infant botulism cases. This is the first reported case of infant botulism due to C. botulinum type Af worldwide.

  13. Characterization of Clostridium sp. RKD producing botulinum-like neurotoxin.

    Science.gov (United States)

    Dixit, Aparna; Dhaked, Ram Kumar; Alam, Syed Imteyaz; Singh, Lokendra

    2005-07-01

    A Gram positive, motile, rod-shaped, strictly anaerobic bacterium isolated from intestine of decaying fish was identified as Clostridium sp. RKD and produced a botulinum type B-like neurotoxin as suggested by mouse bioassay and protection with anti botulinum antibodies. The neurotoxicity was functionally characterized by the phrenic nerve hemi-diaphragm assay. Phylogenetic analysis based on 16S rDNA sequence, placed it at a different position from the reported strains of Clostridium botulinum. The strain exhibited differences from both Clostridium botulinum and Clostridium tetani with respect to morphological, biochemical and chemotaxonomic characteristics. Botulinum group specific and serotype specific primers amplified the DNA fragments of 260 and 727 bp, respectively, indicating presence of botulinum type 'B' toxin gene. Sequence of nearly 700 bp amplified using primers specific for botulinum neurotoxin type B gene, did not show any significant match in the database when subjected to BLAST search.

  14. Clostridium botulinum strains producing BoNT/F4 or BoNT/F5.

    Science.gov (United States)

    Raphael, Brian H; Bradshaw, Marite; Kalb, Suzanne R; Joseph, Lavin A; Lúquez, Carolina; Barr, John R; Johnson, Eric A; Maslanka, Susan E

    2014-05-01

    Botulinum neurotoxin type F (BoNT/F) may be produced by Clostridium botulinum alone or in combination with another toxin type such as BoNT/A or BoNT/B. Type F neurotoxin gene sequences have been further classified into seven toxin subtypes. Recently, the genome sequence of one strain of C. botulinum (Af84) was shown to contain three neurotoxin genes (bont/F4, bont/F5, and bont/A2). In this study, eight strains containing bont/F4 and seven strains containing bont/F5 were examined. Culture supernatants produced by these strains were incubated with BoNT/F-specific peptide substrates. Cleavage products of these peptides were subjected to mass spectral analysis, allowing detection of the BoNT/F subtypes present in the culture supernatants. PCR analysis demonstrated that a plasmid-specific marker (PL-6) was observed only among strains containing bont/F5. Among these strains, Southern hybridization revealed the presence of an approximately 242-kb plasmid harboring bont/F5. Genome sequencing of four of these strains revealed that the genomic backgrounds of strains harboring either bont/F4 or bont/F5 are diverse. None of the strains analyzed in this study were shown to produce BoNT/F4 and BoNT/F5 simultaneously, suggesting that strain Af84 is unusual. Finally, these data support a role for the mobility of a bont/F5-carrying plasmid among strains of diverse genomic backgrounds.

  15. Isolation and functional characterization of the novel Clostridium botulinum neurotoxin A8 subtype.

    Science.gov (United States)

    Kull, Skadi; Schulz, K Melanie; Weisemann, Jasmin; Kirchner, Sebastian; Schreiber, Tanja; Bollenbach, Alexander; Dabrowski, P Wojtek; Nitsche, Andreas; Kalb, Suzanne R; Dorner, Martin B; Barr, John R; Rummel, Andreas; Dorner, Brigitte G

    2015-01-01

    Botulism is a severe neurological disease caused by the complex family of botulinum neurotoxins (BoNT). Based on the different serotypes known today, a classification of serotype variants termed subtypes has been proposed according to sequence diversity and immunological properties. However, the relevance of BoNT subtypes is currently not well understood. Here we describe the isolation of a novel Clostridium botulinum strain from a food-borne botulism outbreak near Chemnitz, Germany. Comparison of its botulinum neurotoxin gene sequence with published sequences identified it to be a novel subtype within the BoNT/A serotype designated BoNT/A8. The neurotoxin gene is located within an ha-orfX+ cluster and showed highest homology to BoNT/A1, A2, A5, and A6. Unexpectedly, we found an arginine insertion located in the HC domain of the heavy chain, which is unique compared to all other BoNT/A subtypes known so far. Functional characterization revealed that the binding characteristics to its main neuronal protein receptor SV2C seemed unaffected, whereas binding to membrane-incorporated gangliosides was reduced in comparison to BoNT/A1. Moreover, we found significantly lower enzymatic activity of the natural, full-length neurotoxin and the recombinant light chain of BoNT/A8 compared to BoNT/A1 in different endopeptidase assays. Both reduced ganglioside binding and enzymatic activity may contribute to the considerably lower biological activity of BoNT/A8 as measured in a mouse phrenic nerve hemidiaphragm assay. Despite its reduced activity the novel BoNT/A8 subtype caused severe botulism in a 63-year-old male. To our knowledge, this is the first description and a comprehensive characterization of a novel BoNT/A subtype which combines genetic information on the neurotoxin gene cluster with an in-depth functional analysis using different technical approaches. Our results show that subtyping of BoNT is highly relevant and that understanding of the detailed toxin function might

  16. Isolation and functional characterization of the novel Clostridium botulinum neurotoxin A8 subtype.

    Directory of Open Access Journals (Sweden)

    Skadi Kull

    Full Text Available Botulism is a severe neurological disease caused by the complex family of botulinum neurotoxins (BoNT. Based on the different serotypes known today, a classification of serotype variants termed subtypes has been proposed according to sequence diversity and immunological properties. However, the relevance of BoNT subtypes is currently not well understood. Here we describe the isolation of a novel Clostridium botulinum strain from a food-borne botulism outbreak near Chemnitz, Germany. Comparison of its botulinum neurotoxin gene sequence with published sequences identified it to be a novel subtype within the BoNT/A serotype designated BoNT/A8. The neurotoxin gene is located within an ha-orfX+ cluster and showed highest homology to BoNT/A1, A2, A5, and A6. Unexpectedly, we found an arginine insertion located in the HC domain of the heavy chain, which is unique compared to all other BoNT/A subtypes known so far. Functional characterization revealed that the binding characteristics to its main neuronal protein receptor SV2C seemed unaffected, whereas binding to membrane-incorporated gangliosides was reduced in comparison to BoNT/A1. Moreover, we found significantly lower enzymatic activity of the natural, full-length neurotoxin and the recombinant light chain of BoNT/A8 compared to BoNT/A1 in different endopeptidase assays. Both reduced ganglioside binding and enzymatic activity may contribute to the considerably lower biological activity of BoNT/A8 as measured in a mouse phrenic nerve hemidiaphragm assay. Despite its reduced activity the novel BoNT/A8 subtype caused severe botulism in a 63-year-old male. To our knowledge, this is the first description and a comprehensive characterization of a novel BoNT/A subtype which combines genetic information on the neurotoxin gene cluster with an in-depth functional analysis using different technical approaches. Our results show that subtyping of BoNT is highly relevant and that understanding of the detailed

  17. Occurrence of human pathogenic Clostridium botulinum among healthy dairy animals: an emerging public health hazard.

    Science.gov (United States)

    Abdel-Moein, Khaled A; Hamza, Dalia A

    2016-01-01

    The current study was conducted to investigate the occurrence of human pathogenic Clostridium botulinum in the feces of dairy animals. Fecal samples were collected from 203 apparently healthy dairy animals (50 cattle, 50 buffaloes, 52 sheep, 51 goats). Samples were cultured to recover C. botulinum while human pathogenic C. botulinum strains were identified after screening of all C. botulinum isolates for the presence of genes that encode toxins type A, B, E, F. The overall prevalence of C. botulinum was 18.7% whereas human pathogenic C. botulinum strains (only type A) were isolated from six animals at the rates of 2, 2, 5.8, and 2% for cattle, buffaloes, sheep, and goats, respectively. High fecal carriage rates of C. botulinum among apparently healthy dairy animals especially type A alarm both veterinary and public health communities for a potential role which may be played by dairy animals in the epidemiology of such pathogen.

  18. Purification and characterization of neurotoxin complex from a dual toxin gene containing Clostridium Botulinum Strain PS-5.

    Science.gov (United States)

    Singh, Ajay K; Sachdeva, Amita; Degrasse, Jeffrey A; Croley, Timothy R; Stanker, Larry H; Hodge, David; Sharma, Shashi K

    2013-04-01

    Botulinum neurotoxins are produced as a toxin complex (TC) which consists of neurotoxin (NT) and neurotoxin associated proteins. The characterization of NT in its native state is an essential step for developing diagnostics and therapeutic countermeasures against botulism. The presence of NT genes was validated by PCR amplification of toxin specific fragments from genomic DNA of Clostridium botulinum strain PS-5 which indicated the presence of both serotype A and B genes on PS-5 genome. Further, TC was purified and characterized by Western blotting, Digoxin-enzyme linked immunosorbent assay, endopeptidase activity assay, and Liquid chromatography-Mass spectrometry. The data showed the presence of serotype A specific neurotoxin. Based on the analysis of neurotoxin genes and characterization of TC, PS-5 strain appears as a serotype A (B) strain of C. botulinum which produces only serotype A specific TC in the cell culture medium.

  19. Clostridium botulinum strain Af84 contains three neurotoxin gene clusters: bont/A2, bont/F4 and bont/F5.

    Directory of Open Access Journals (Sweden)

    Nir Dover

    Full Text Available Sanger and shotgun sequencing of Clostridium botulinum strain Af84 type Af and its botulinum neurotoxin gene (bont clusters identified the presence of three bont gene clusters rather than the expected two. The three toxin gene clusters consisted of bont subtypes A2, F4 and F5. The bont/A2 and bont/F4 gene clusters were located within the chromosome (the latter in a novel location, while the bont/F5 toxin gene cluster was located within a large 246 kb plasmid. These findings are the first identification of a C. botulinum strain that contains three botulinum neurotoxin gene clusters.

  20. Construction of "Toxin Complex" in a Mutant Serotype C Strain of Clostridium botulinum Harboring a Defective Neurotoxin Gene.

    Science.gov (United States)

    Suzuki, Tomonori; Nagano, Thomas; Niwa, Koichi; Uchino, Masataka; Tomizawa, Motohiro; Sagane, Yoshimasa; Watanabe, Toshihiro

    2017-01-01

    A non-toxigenic mutant of the toxigenic serotype C Clostridium botulinum strain Stockholm (C-St), C-N71, does not produce the botulinum neurotoxin (BoNT). However, the original strain C-St produces botulinum toxin complex, in which BoNT is associated with non-toxic non-hemagglutinin (NTNHA) and three hemagglutinin proteins (HA-70, HA-33, and HA-17). Therefore, in this study, we aimed to elucidate the effects of bont gene knockout on the formation of the "toxin complex." Nucleotide sequence analysis revealed that a premature stop codon was introduced in the bont gene, whereas other genes were not affected by this mutation. Moreover, we successfully purified the "toxin complex" produced by C-N71. The "toxin complex" was identified as a mixture of NTNHA/HA-70/HA-17/HA-33 complexes with intact NTNHA or C-terminally truncated NTNHA, without BoNT. These results indicated that knockout of the bont gene does not affect the formation of the "toxin complex." Since the botulinum toxin complex has been shown to play an important role in oral toxin transport in the human and animal body, a non-neurotoxic "toxin complex" of C-N71 may be valuable for the development of an oral drug delivery system.

  1. Meta-analysis of D-values of proteolytic Clostridium botulinum and its surrogate strain Clostridium sporogenes PA 3679.

    Science.gov (United States)

    Diao, Mamadou Moctar; André, Stéphane; Membré, Jeanne-Marie

    2014-03-17

    Foodborne botulism is a serious disease resulting from ingestion of preformed Clostridium botulinum neurotoxin in foodstuff. Since the 19th century, the heat resistance of this spore forming bacteria has been extensively studied in order to guarantee the public health associated with low acidic, ambient stable products. The most largely used heat resistance parameters in thermal settings of such products are the D121.1°C values (time required to have a 10-fold decrease of the spore count, at 121.1°C) and the z-values (temperature increase to have a 10-fold decrease of D-values). To determine D121.1°C and z-values of proteolytic C. botulinum and its nontoxigenic surrogate strain C. sporogenes PA3679, a dataset of 911 D-values was collected from 38 scientific studies. Within a meta-analysis framework, a mixed-effect linear model was developed with the log D-value (min) as response and the heat treatment temperature as explicative variable. The studies (38), the C. botulinum strains (11), and the heat treatment media (liquid media and various food matrices, split into nine categories in total) were considered as co-variables having a random effect. The species (C. botulinum and C. sporogenes) and the pH (five categories) were considered as co-variables having a fixed effect. Overall, the model gave satisfactory results with a residual standard deviation of 0.22. The heat resistance of proteolytic C. botulinum was found significantly lower than the C. sporogenes PA 3679 one: the mean D-values at the reference temperature of 121.1°C, in liquid media and pH neutral, were estimated to 0.19 and 1.28min for C. botulinum and C. sporogenes, respectively. On the other hand, the mean z-values of the two species were similar: 11.3 and 11.1°C for C. botulinum and C. sporogenes, respectively. These results will be applied to thermal settings of low-acid ambient stable products.

  2. Characterization of the spore surface and exosporium proteins of Clostridium sporogenes; implications for Clostridium botulinum group I strains.

    Science.gov (United States)

    Janganan, Thamarai K; Mullin, Nic; Tzokov, Svetomir B; Stringer, Sandra; Fagan, Robert P; Hobbs, Jamie K; Moir, Anne; Bullough, Per A

    2016-10-01

    Clostridium sporogenes is a non-pathogenic close relative and surrogate for Group I (proteolytic) neurotoxin-producing Clostridium botulinum strains. The exosporium, the sac-like outermost layer of spores of these species, is likely to contribute to adhesion, dissemination, and virulence. A paracrystalline array, hairy nap, and several appendages were detected in the exosporium of C. sporogenes strain NCIMB 701792 by EM and AFM. The protein composition of purified exosporium was explored by LC-MS/MS of tryptic peptides from major individual SDS-PAGE-separated protein bands, and from bulk exosporium. Two high molecular weight protein bands both contained the same protein with a collagen-like repeat domain, the probable constituent of the hairy nap, as well as cysteine-rich proteins CsxA and CsxB. A third cysteine-rich protein (CsxC) was also identified. These three proteins are also encoded in C. botulinum Prevot 594, and homologues (75-100% amino acid identity) are encoded in many other Group I strains. This work provides the first insight into the likely composition and organization of the exosporium of Group I C. botulinum spores.

  3. Effects of Megaplasmid Loss on Growth of Neurotoxigenic Clostridium butyricum Strains and Botulinum Neurotoxin Type E Expression.

    Science.gov (United States)

    Scalfaro, Concetta; Iacobino, Angelo; Grande, Laura; Morabito, Stefano; Franciosa, Giovanna

    2016-01-01

    Clostridium butyricum strains that atypically produce the botulinum neurotoxin type E (BoNT/E) possess a megaplasmid of unknown functions in their genome. In this study, we cured two botulinum neurotoxigenic C. butyricum type E strains of their megaplasmids, and compared the obtained megaplasmid-cured strains to their respective wild-type parental strains. Our results showed that the megaplasmids do not confer beta-lactam resistance on the neurotoxigenic C. butyricum type E strains, although they carry several putative beta-lactamase genes. Instead, we found that the megaplasmids are essential for growth of the neurotoxigenic C. butyricum type E strains at the relatively low temperature of 15°C, and are also relevant for growth of strains under limiting pH and salinity conditions, as well as under favorable environmental conditions. Moreover, the presence of the megaplasmids was associated with increased transcript levels of the gene encoding BoNT/E in the C. butyricum type E strains, indicating that the megaplasmids likely contain transcriptional regulators. However, the levels of BoNT/E in the supernatants of the cured and uncured strains were similar after 24 and 48 h culture, suggesting that expression of BoNT/E in the C. butyricum type E strains is not ultimately controlled by the megaplasmids. Together, our results reveal that the C. butyricum type E megaplasmids exert pleiotropic effects on the growth of their microbial hosts under optimal and limiting environmental conditions, and also highlight the possibility of original regulatory mechanisms controlling the expression of BoNT/E.

  4. Plasmidome interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum converts strains of independent lineages into distinctly different pathogens.

    Science.gov (United States)

    Skarin, Hanna; Segerman, Bo

    2014-01-01

    Clostridium botulinum (group III), Clostridium novyi and Clostridium haemolyticum are well-known pathogens causing animal botulism, gas gangrene/black disease, and bacillary hemoglobinuria, respectively. A close genetic relationship exists between the species, which has resulted in the collective term C. novyi sensu lato. The pathogenic traits in these species, e.g., the botulinum neurotoxin and the novyi alpha toxin, are mainly linked to a large plasmidome consisting of plasmids and circular prophages. The plasmidome of C. novyi sensu lato has so far been poorly characterized. In this study we explored the genomic relationship of a wide range of strains of C. novyi sensu lato with a special focus on the dynamics of the plasmidome. Twenty-four genomes were sequenced from strains selected to represent as much as possible the genetic diversity in C. novyi sensu lato. Sixty-one plasmids were identified in these genomes and 28 of them were completed. The genomic comparisons revealed four separate lineages, which did not strictly correlate with the species designations. The plasmids were categorized into 13 different plasmid groups on the basis of their similarity and conservation of plasmid replication or partitioning genes. The plasmid groups, lineages and species were to a large extent entwined because plasmids and toxin genes had moved across the lineage boundaries. This dynamic process appears to be primarily driven by phages. We here present a comprehensive characterization of the complex species group C. novyi sensu lato, explaining the intermixed genetic properties. This study also provides examples how the reorganization of the botulinum toxin and the novyi alpha toxin genes within the plasmidome has affected the pathogenesis of the strains.

  5. Plasmidome interchange between Clostridium botulinum, Clostridium novyi and Clostridium haemolyticum converts strains of independent lineages into distinctly different pathogens.

    Directory of Open Access Journals (Sweden)

    Hanna Skarin

    Full Text Available Clostridium botulinum (group III, Clostridium novyi and Clostridium haemolyticum are well-known pathogens causing animal botulism, gas gangrene/black disease, and bacillary hemoglobinuria, respectively. A close genetic relationship exists between the species, which has resulted in the collective term C. novyi sensu lato. The pathogenic traits in these species, e.g., the botulinum neurotoxin and the novyi alpha toxin, are mainly linked to a large plasmidome consisting of plasmids and circular prophages. The plasmidome of C. novyi sensu lato has so far been poorly characterized. In this study we explored the genomic relationship of a wide range of strains of C. novyi sensu lato with a special focus on the dynamics of the plasmidome. Twenty-four genomes were sequenced from strains selected to represent as much as possible the genetic diversity in C. novyi sensu lato. Sixty-one plasmids were identified in these genomes and 28 of them were completed. The genomic comparisons revealed four separate lineages, which did not strictly correlate with the species designations. The plasmids were categorized into 13 different plasmid groups on the basis of their similarity and conservation of plasmid replication or partitioning genes. The plasmid groups, lineages and species were to a large extent entwined because plasmids and toxin genes had moved across the lineage boundaries. This dynamic process appears to be primarily driven by phages. We here present a comprehensive characterization of the complex species group C. novyi sensu lato, explaining the intermixed genetic properties. This study also provides examples how the reorganization of the botulinum toxin and the novyi alpha toxin genes within the plasmidome has affected the pathogenesis of the strains.

  6. Insights into the evolutionary origins of clostridial neurotoxins from analysis of the Clostridium botulinum strain A neurotoxin gene cluster

    Directory of Open Access Journals (Sweden)

    Meiering Elizabeth M

    2008-11-01

    Full Text Available Abstract Background Clostridial neurotoxins (CNTs are the most deadly toxins known and causal agents of botulism and tetanus neuroparalytic diseases. Despite considerable progress in understanding CNT structure and function, the evolutionary origins of CNTs remain a mystery as they are unique to Clostridium and possess a sequence and structural architecture distinct from other protein families. Uncovering the origins of CNTs would be a significant contribution to our understanding of how pathogens evolve and generate novel toxin families. Results The C. botulinum strain A genome was examined for potential homologues of CNTs. A key link was identified between the neurotoxin and the flagellin gene (CBO0798 located immediately upstream of the BoNT/A neurotoxin gene cluster. This flagellin sequence displayed the strongest sequence similarity to the neurotoxin and NTNH homologue out of all proteins encoded within C. botulinum strain A. The CBO0798 gene contains a unique hypervariable region, which in closely related flagellins encodes a collagenase-like domain. Remarkably, these collagenase-containing flagellins were found to possess the characteristic HEXXH zinc-protease motif responsible for the neurotoxin's endopeptidase activity. Additional links to collagenase-related sequences and functions were detected by further analysis of CNTs and surrounding genes, including sequence similarities to collagen-adhesion domains and collagenases. Furthermore, the neurotoxin's HCRn domain was found to exhibit both structural and sequence similarity to eukaryotic collagen jelly-roll domains. Conclusion Multiple lines of evidence suggest that the neurotoxin and adjacent genes evolved from an ancestral collagenase-like gene cluster, linking CNTs to another major family of clostridial proteolytic toxins. Duplication, reshuffling and assembly of neighboring genes within the BoNT/A neurotoxin gene cluster may have lead to the neurotoxin's unique architecture. This

  7. Genetic diversity of the flagellin genes of Clostridium botulinum groups I and II.

    Science.gov (United States)

    Woudstra, Cedric; Lambert, Dominic; Anniballi, Fabrizio; De Medici, Dario; Austin, John; Fach, Patrick

    2013-07-01

    Botulinum neurotoxins (BoNTs) are produced by phenotypically and genetically different Clostridium species, including Clostridium botulinum and some strains of Clostridium baratii (serotype F) and Clostridium butyricum (serotype E). BoNT-producing clostridia responsible for human botulism encompass strains of group I (secreting proteases, producing toxin serotype A, B, or F, and growing optimally at 37°C) and group II (nonproteolytic, producing toxin serotype E, B, or F, and growing optimally at 30°C). Here we report the development of real-time PCR assays for genotyping C. botulinum strains of groups I and II based on flaVR (variable region sequence of flaA) sequences and the flaB gene. Real-time PCR typing of regions flaVR1 to flaVR10 and flaB was optimized and validated with 62 historical and Canadian C. botulinum strains that had been previously typed. Analysis of 210 isolates of European origin allowed the identification of four new C. botulinum flaVR types (flaVR11 to flaVR14) and one new flaVR type specific to C. butyricum type E (flaVR15). The genetic diversity of the flaVR among C. botulinum strains investigated in the present study reveals the clustering of flaVR types into 5 major subgroups. Subgroups 1, 3, and 4 contain proteolytic Clostridium botulinum, subgroup 2 is made up of nonproteolytic C. botulinum only, and subgroup 5 is specific to C. butyricum type E. The genetic variability of the flagellin genes carried by C. botulinum and the possible association of flaVR types with certain geographical areas make gene profiling of flaVR and flaB promising in molecular surveillance and epidemiology of C. botulinum.

  8. Characterisation of non-toxigenic Clostridium spp. strains, to use as surrogates for non-proteolytic Clostridium botulinum in chilled food challenge testing.

    Science.gov (United States)

    Parker, M D; Barrett, P I; Shepherd, J; Price, L J; Bull, S D

    2015-01-01

    Under many of the conditions studied, a two-strain cocktail of non-toxigenic Clostridium spp. was found to be suitable as a surrogate for non-proteolytic Clostridium botulinum, and has the potential for use in chilled food challenge tests measuring growth. Non-toxigenic surrogates could also be used in thermal process screening studies.

  9. Clostridium botulinum group I strain genotyping by 15-locus multilocus variable-number tandem-repeat analysis

    NARCIS (Netherlands)

    Fillo, S.; Giordani, F.; Anniballi, F.; Gorgé, O.; Ramisse, V.; Vergnaud, G.; Riehm, J.M.; Scholz, H.C.; Splettstoesser, W.D.; Kieboom, J.; Olsen, J.-S.; Fenicia, L.; Lista, F.

    2011-01-01

    Clostridium botulinum is a taxonomic designation that encompasses a broad variety of spore-forming, Gram-positive bacteria producing the botulinum neurotoxin (BoNT). C. botulinum is the etiologic agent of botulism, a rare but severe neuroparalytic disease. Fine-resolution genetic characterization of

  10. 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)

  11. Isolation and Identification of Clostridium botulinum in Production Environment of Bean Products%新疆豆制品生产环境中肉毒梭菌的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    刘凯; 卢士玲; 李开雄

    2011-01-01

    从新疆伊犁察布查尔县的土壤中分离到1株优势菌,该分离株生长特性与A型肉毒梭菌(Clostridiumbotulinum type A)一致,为革兰氏阳性粗大杆菌,在EYA培养基上菌落边缘呈锯齿状,表面粗糙不规则,有较大的乳浊环。根据分离株的形态、生理生化特性,结合琼脂糖凝胶凝胶电泳图像、16SrDNA V6-V8序列(GenBank登录号:JN248616)与系统发育树分析,将其鉴定为A型肉毒梭菌。%The dominant strain(GenBank accession number: JN248616) was isolated from the soil in Qapqal County,Xinjiang.Its growth characteristics were consistent with Clostridium botulinum of type A which belonged to thick gram-positive bacillus.Colonies in EYA culture medium showed serrated edge and irregular rough surface,with large opaque rings.Based on morphology of the strain,characteristics of physical and biochemistry,and combined with gel electrophoresis,16S rDNA sequences and phylogenetic tree,the strain was identified as Clostridium botulinum of type A.

  12. Genetic diversity within Clostridium botulinum serotypes, botulinum neurotoxin gene clusters and toxin subtypes.

    Science.gov (United States)

    Hill, Karen K; Smith, Theresa J

    2013-01-01

    Clostridium botulinum is a species of spore-forming anaerobic bacteria defined by the expression of any one or two of seven serologically distinct botulinum neurotoxins (BoNTs) designated BoNT/A-G. This Gram-positive bacterium was first identified in 1897 and since then the paralyzing and lethal effects of its toxin have resulted in the recognition of different forms of the intoxication known as food-borne, infant, or wound botulism. Early microbiological and biochemical characterization of C. botulinum isolates revealed that the bacteria within the species had different characteristics and expressed different toxin types. To organize the variable bacterial traits within the species, Group I-IV designations were created. Interestingly, it was observed that isolates within different Groups could express the same toxin type and conversely a single Group could express different toxin types. This discordant phylogeny between the toxin and the host bacteria indicated that horizontal gene transfer of the toxin was responsible for the variation observed within the species. The recent availability of multiple C. botulinum genomic sequences has offered the ability to bioinformatically analyze the locations of the bont genes, the composition of their toxin gene clusters, and the genes flanking these regions to understand their variation. Comparison of the genomic sequences representing multiple serotypes indicates that the bont genes are not in random locations. Instead the analyses revealed specific regions where the toxin genes occur within the genomes representing serotype A, B, C, E, and F C. botulinum strains and C. butyricum type E strains. The genomic analyses have provided evidence of horizontal gene transfer, site-specific insertion, and recombination events. These events have contributed to the variation observed among the neurotoxins, the toxin gene clusters and the bacteria that contain them, and has supported the historical microbiological, and biochemical

  13. Botulinum Toxin; Bioterror and Biomedicinal Agent

    Directory of Open Access Journals (Sweden)

    Jiri Patocka

    2006-04-01

    Full Text Available Botulinum toxin is a group of seven homologous, highly poisonous proteins isolated fromfermentation of the anaerobic bacterium Clostridium botulinum, which naturally occurs in soiland can grow on many meats and vegetables. Botulinum toxin causes neuromuscular disordercalled botulism, which is a potentially lethal disease. There are three types of botulism: Food,wound, and infant botulism. It can lead to death unless appropriate therapy is done. Due to theseverity and potency of botulinum toxin, its importance as a biological weapon is of majorconcern to public health officials. Nevertheless, botulinum toxin is also medicament.

  14. Isolation of a Bacterium Strain Degraded Agar

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    One in 58 strains of bacteria isolated from the compost showed clear colonies after a few days of growth on the plates containing medium made of only agar and water.Water suspension contained only agar (2 and 8g·L -1 ) with two controls (normal saline,LB medium) was inoculated with the bacterium BR5-1 to see whether there was an increasement of the alive bacteria concentration after 48 h of the growth.The results showed that there was a significant rising of the alive bacteria concentration in the agar susp...

  15. Quantitative real-time reverse transcription-PCR analysis reveals stable and prolonged neurotoxin cluster gene activity in a Clostridium botulinum type E strain at refrigeration temperature.

    Science.gov (United States)

    Chen, Ying; Korkeala, Hannu; Lindén, Jere; Lindström, Miia

    2008-10-01

    The relative expression levels of six botulinum neurotoxin cluster genes in a group II Clostridium botulinum type E strain grown at 10 or 30 degrees C were investigated using quantitative real-time reverse transcription-PCR. An enzyme-linked immunosorbent assay was used to confirm neurotoxin expression. Distinct mRNA and toxin production patterns were observed at the two temperatures. The average relative mRNA levels at 10 degrees C were higher than (ntnh and p47), similar to (botE), or lower than (orfx1, orfx2, orfx3) those at 30 degrees C. The maximum botE expression levels and average neurotoxin levels at 10 degrees C were 45 to 65% of those at 30 degrees C. The relative mRNA levels at 10 degrees C declined generally slowly within 8 days, as opposed to the rapid decline observed at 30 degrees C within 24 h. Distinct expression patterns of the six genes at the two temperatures suggest that the type E neurotoxin cluster genes are transcribed as two tricistronic operons at 30 degrees C, whereas at 10 degrees C monocistronic (botE or orfx1 alone) and bicistronic (ntnh-p47 and orfx2-orfx3) transcription may dominate. Thus, type E botulinum neurotoxin production may be involved with various temperature-dependent regulatory events. In light of group II C. botulinum type E being a dangerous food-borne pathogen, these findings may be important in terms of the safety of refrigerated packaged foods of extended durability.

  16. Isolation and Characterization of a New Arthrospira Strain

    OpenAIRE

    2008-01-01

    A filamentous microorganism, morphologically similar to the cyanobacterium Arthrospira, was isolated from Mangueira Lagoon in Brazil, from which Arthrospira has not previously been isolated. Random amplified polymorphic DNA (RAPD) comparison with the standard Arthrospira platensis strains LEB 52 and Paracas indicated that the organism isolated was an Arthrospira isolate, which we denominated strain LEB 18. The RAPD analysis showed conserved sequences which indicated that the three strains bel...

  17. New Insights into the Genetic Diversity of Clostridium botulinum Group III through Extensive Genome Exploration.

    Science.gov (United States)

    Woudstra, Cédric; Le Maréchal, Caroline; Souillard, Rozenn; Bayon-Auboyer, Marie-Hélène; Mermoud, Isabelle; Desoutter, Denise; Fach, Patrick

    2016-01-01

    Animal botulism is caused by group III Clostridium botulinum strains producing type C and D toxins, or their chimeric forms C/D and D/C. Animal botulism is considered an emerging disease in Europe, notably in poultry production. Before our study, 14 genomes from different countries were available in the public database, but none were from France. In order to investigate the genetic relationship of French strains with different geographical areas and find new potential typing targets, 17 strains of C. botulinum group III were sequenced (16 from France and one from New Caledonia). Fourteen were type C/D strains isolated from chickens, ducks, guinea fowl and turkeys and three were type D/C strains isolated from cattle. The New Caledonian strain was a type D/C strain. Whole genome sequence analysis showed the French strains to be closely related to European strains from C. botulinum group III lineages Ia and Ib. The investigation of CRISPR sequences as genetic targets for differentiating strains in group III proved to be irrelevant for type C/D due to a deficient CRISPR/Cas mechanism, but not for type D/C. Conversely, the extrachromosomal elements of type C/D strains could be used to generate a genetic ID card. The highest level of discrimination was achieved with SNP core phylogeny, which allowed differentiation up to strain level and provide the most relevant information for genetic epidemiology studies and discrimination.

  18. New Insights into the genetic diversity of Clostridium botulinum Group III through extensive genome exploration

    Directory of Open Access Journals (Sweden)

    Cédric eWoudstra

    2016-05-01

    Full Text Available Animal botulism is caused by group III Clostridium botulinum strains producing type C and D toxins, or their chimeric forms C/D and D/C. Animal botulism is considered an emerging disease in Europe, notably in poultry production. Before our study, 14 genomes from different countries were available in the public database, but none were from France. In order to investigate the genetic relationship of French strains with different geographical areas and find new potential typing targets, 17 strains of C. botulinum group III were sequenced (16 from France and one from New Caledonia. Fourteen were type C/D strains isolated from chickens, ducks, guinea fowl and turkeys and three were type D/C strains isolated from cattle. The New Caledonian strain was a type D/C strain. Whole genome sequence analysis showed the French strains to be closely related to European strains from C. botulinum group III lineages Ia and Ib. The investigation of CRISPR sequences as genetic targets for differentiating strains in group III proved to be irrelevant for type C/D due to a deficient CRISPR/Cas mechanism, but not for type D/C. Conversely, the extrachromosomal elements of type C/D strains could be used to generate a genetic ID card. The highest level of discrimination was achieved with SNP core phylogeny, which allowed differentiation up to strain level and provide the most relevant information for genetic epidemiology studies and discrimination.

  19. Botulinum toxin

    Directory of Open Access Journals (Sweden)

    Nigam P

    2010-01-01

    Full Text Available Botulinum toxin, one of the most poisonous biological substances known, is a neurotoxin produced by the bacterium Clostridium botulinum. C. botulinum elaborates eight antigenically distinguishable exotoxins (A, B, C 1 , C 2 , D, E, F and G. All serotypes interfere with neural transmission by blocking the release of acetylcholine, the principal neurotransmitter at the neuromuscular junction, causing muscle paralysis. The weakness induced by injection with botulinum toxin A usually lasts about three months. Botulinum toxins now play a very significant role in the management of a wide variety of medical conditions, especially strabismus and focal dystonias, hemifacial spasm, and various spastic movement disorders, headaches, hypersalivation, hyperhidrosis, and some chronic conditions that respond only partially to medical treatment. The list of possible new indications is rapidly expanding. The cosmetological applications include correction of lines, creases and wrinkling all over the face, chin, neck, and chest to dermatological applications such as hyperhidrosis. Injections with botulinum toxin are generally well tolerated and side effects are few. A precise knowledge and understanding of the functional anatomy of the mimetic muscles is absolutely necessary to correctly use botulinum toxins in clinical practice.

  20. A unique restriction site in the flaA gene allows rapid differentiation of group I and group II Clostridium botulinum strains by PCR-restriction fragment length polymorphism analysis.

    Science.gov (United States)

    Paul, Catherine J; Tran, Shulin; Tam, Kevin J; Austin, John W

    2007-09-01

    Clostridium botulinum produces the potent botulinum neurotoxin, the causative agent of botulism. Based on distinctive physiological traits, strains of C. botulinum can be divided into four groups: however, only groups I and II are associated with human illness. Alignment of the flaA gene sequences from 40 group I and 40 group II strains identified a single BsrG1 restriction cut site that was present at base pair 283 in all group II flaA sequences and was not found in any group I sequence. The flaA gene was amplified by rapid colony PCR from 22 group I strains and 18 group II strains and digested with BsrGI restriction enzyme. Standard agarose gel electrophoresis with ethidium bromide staining showed two fragments, following restriction digestion of group II flaA gene amplicons with BsrGI, but only a single band of uncut flaA from group I strains. Combining rapid colony PCR with BsrGI restriction digest of the flaA gene at 60 degrees C is a significant improvement over current methods, such as meat digestion or amplified fragment length polymorphism, as a strain can be identified as either group I or group II in under 5 h when starting with a visible plated C. botulinum colony.

  1. Ciprofloxacin susceptibility reduction of Salmonella strains isolated from outbreaks

    Directory of Open Access Journals (Sweden)

    Roberta B. Souza

    2010-06-01

    Full Text Available The antimicrobial susceptibility of 212 Salmonella strains isolated from patients and foods was evaluated and 45% were found to be resistant to nalidixic acid. Nalidixic acid resistant strains showed a higher minimal inhibitory concentration for ciprofloxacin than sensitive strains. During the study an increase of strains with reduced susceptibility to ciprofloxacin was also observed.

  2. Historical Perspectives and Guidelines for Botulinum Neurotoxin Subtype Nomenclature

    Science.gov (United States)

    2016-08-26

    20National Institute for Biological Standards and Control, a centre of Medicines and Healthcare Products Regulatory Agency, Hertfordshire UK. 21Botulinum...The technology resulting in the production of monoclonal antibodies provided a new way of evaluating these toxins serologically. Monoclonal...botulinum neurotoxin-producing strains representing three clostridial species illustrate the mobility and diversity of botulinum neurotoxin genes, Infect

  3. Isolation of botulinolysin, a thiol-activated hemolysin, from serotype D Clostridium botulinum: A species-specific gene duplication in Clostridia.

    Science.gov (United States)

    Suzuki, Tomonori; Nagano, Thomas; Niwa, Koichi; Mutoh, Shingo; Uchino, Masataka; Tomizawa, Motohiro; Sagane, Yoshimasa; Watanabe, Toshihiro

    2016-12-01

    Botulinolysin (BLY) is a toxin produced by Clostridium botulinum that belongs to a group of thiol-activated hemolysins. In this study, a protein exhibiting hemolytic activity was purified from the culture supernatant of C. botulinum serotype D strain 4947. The purified protein displayed a single band by sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular mass of 55kDa, and its N-terminal and internal amino acid sequences exhibited high similarity to a group of thiol-activated hemolysins produced by gram-positive bacteria. Thus, the purified protein was identified as the BLY. Using the nucleotide sequences of previously cloned genes for hemolysins, two types of genes encoding BLY-like proteins were cloned unexpectedly. Molecular modeling analysis indicated that the products of both genes displayed very similar structures, despite the low sequence similarity. In silico screening revealed a specific duplication of the hemolysin gene restricted to serotypes C and D of C. botulinum and their related species among thiol-activated hemolysin-producing bacteria. Our findings provide important insights into the genetic characteristics of pathogenic bacteria.

  4. Alternative sigma factor SigK has a role in stress tolerance of group I Clostridium botulinum strain ATCC 3502.

    Science.gov (United States)

    Dahlsten, Elias; Kirk, David; Lindström, Miia; Korkeala, Hannu

    2013-06-01

    The role of the alternative sigma factor SigK in cold and osmotic stress tolerance of Clostridium botulinum ATCC 3502 was demonstrated by induction of sigK after temperature downshift and exposure to hyperosmotic conditions and by impaired growth of the sigK mutants under the respective conditions.

  5. Isolation of clinical strains of Pseudomonas aeruginosa harboring different plasmids.

    Science.gov (United States)

    Ranjbar, R; Owlia, P; Saderi, H; Bameri, Z; Izadi, M; Jonaidi, N; Morovvati, S

    2007-09-01

    Aim of this study was to investigate the presence of plasmids among the strains of P. aeruginosa isolated from clinically diagnosed cases in Tehran in 2006. A total of 38 strains of P. aeruginosa were isolated. With the exception of one isolate, all P. aeruginosa strains harbored at least one plasmid band. The electrophoretic analysis of plasmid DNAs showed different number of plasmid bands among the strains tested. The DNA band of 1.4 kbp was evident in 84.2% of the strains. Approximately 71 and 21% of the isolates harbored concomitantly two and three plasmids, respectively. Isolation of strains with diverse types of plasmids suggests the different cluster of P. aeruginosa might be disseminated during the current study period.

  6. Preliminary study on the isolation of Clostridium butyricum strains from natural sources in the UK and screening the isolates for presence of the type E botulinal toxin gene.

    Science.gov (United States)

    Ghoddusi, Hamid B; Sherburn, Richard

    2010-08-15

    Clostridia such as Clostridium tyrobutyricum, C. pasteurianum and C. butyricum may cause spoilage problems in certain types of food, but they are not normally regarded as dangerous. However some strains of C. butyricum have acquired the type E botulinum neurotoxin gene and have caused both infant and classical botulism in Italy (1986), China (1994) and India (1996). This study was carried out to examine a range of samples from fresh vegetables to food and environmental samples in the UK and test their ability to produce type E botulinal neurotoxin (BoNT) by probing for the presence of the toxin gene. Samples were enriched in modified Bhat and Barker (MBB) broth which is a minimal medium with lactate and acetate as a source of carbon and energy. In addition selective antibiotics are present in the medium to favour the growth of C. butyricum. This was followed by plating out onto iron sulphite agar (ISA) for isolation of C. butyricum from food and environmental samples. A total of 978 samples were tested and 302 (31%) yielded presumptive C. butyricum isolates. The highest percentage of positives came from soil, potato skins, Swede skin, yoghurt and cream. No positive isolates were obtained from pate, garlic or spring greens. A sub-sample of isolates was examined for the presence of gene encoding the type E botulinum neurotoxin using PCR. Only one of the many existing PCR methods was successful and therefore used for screening C. butyricum isolates for the presence of the type E toxin gene. None of the 93 tested isolates were found to be toxigenic (type E botulinal neurotoxin).

  7. Bioactivity characterization of Lactobacillus strains isolated from dairy products

    OpenAIRE

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

    2015-01-01

    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, demonstra...

  8. Analysis of the genetic distribution among members of Clostridium botulinum group I using a novel multilocus sequence typing (MLST) assay.

    Science.gov (United States)

    Olsen, Jaran S; Scholz, Holger; Fillo, Silvia; Ramisse, Vincent; Lista, Florigio; Trømborg, Anette K; Aarskaug, Tone; Thrane, Ingjerd; Blatny, Janet M

    2014-01-01

    Clostridium botulinum is the etiological agent of botulism. Due to food-borne poisoning and the potential use of the extremely toxic botulinum neurotoxin (BoNT) from C. botulinum in bioterror or biocrime related actions, reliable high resolution typing methods for discriminating C. botulinum strains are needed. Partial sequencing of the adk, atpH, gyrB, proC, rpoD and spo0A genes from 51 various C. botulinum/sporogenes isolates was performed, resulting in 37 different sequence types (STs). Analysis of the sequence data revealed a genetic distribution in five larger clusters with a loose correlation to the BoNT serotypes. The developed MLST assay had a slightly lower resolution ability when compared to the MLVA (multilocus variable number of tandem repeat analysis), but the two methods resulted in similar subclusters of the strains possessing the BoNT serotypes A, B and F. The current work presents the development of a novel MLST assay useful for genotyping C. botulinum related to basic phylogenetic research and trace-back analysis in microbial forensic studies.

  9. Botulinum Toxin Therapy

    Science.gov (United States)

    ... care Kids’ zone Video library Find a dermatologist Botulinum toxin therapy Overview Before treatment: This woman disliked her deep frown lines. Botulinum toxin therapy: Overview Also called botulinum rejuvenation Brand names: ...

  10. 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...... strain of M. bovis (PG45(T)) were assayed for variations in the BglII and MfeI restriction sites in the chromosomal DNA by using the amplified fragment length polymorphism (AFLP) fingerprinting technique. The obtained genomic fingerprints consisted of 62-68 AFLP fragments in the size range of 50-500 bp...... 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...

  11. Metabolic activities of Lactobacillus spp. strains isolated from kefir.

    Science.gov (United States)

    Yüksekdag, Zehra Nur; Beyath, Yavuz; Aslim, Belma

    2004-06-01

    A total of 21 strains of Lactobacillus species were isolated from Turkish kefir samples, in order to select the most suitable strains according to their metabolic activities including probiotic properties. As a result of the identification tests, 21 Lactobacillus isolates were identified as L. acidophilus (4%), L. helveticus (9%), L. brevis (9%), L. bulgaricus (14%), L. plantarum (14%), L. casei (19%) and L. lactis (28%). The amount of produced lactic acid, hydrogen peroxide, proteolytic activity, and acetaldehyde productions of Lactobacillus spp. were determined. Different amounts of lactic acid were produced by strains studies; however, lactic acid levels were 1.7-11.4 mg/mL. All strains produced hydrogen peroxide. L. bulgaricus Z14L strain showed no proteolytic activity, L. casei Z6L strain produced the maximum amount (0.16 mg/mL) of proteolytic activity. Acetaldehyde concentration produced in Lactobacillus strains ranged between 0.88-3.52 microg/mL.

  12. Clostridium botulinum Spores Found in Honey from Small Apiaries in Poland

    Directory of Open Access Journals (Sweden)

    Wojtacka Joanna

    2016-12-01

    Full Text Available A total of 102 honey samples collected from small apiaries (≤ 20 hives in Poland were analysed for the presence of Clostridium botulinum spores. The samples were prepared using the dilution centrifugation method and cultured in parallel in cooked meat medium (CMM and tripticase peptone glucose yeast (TPGY enrichment broths. Identification of toxin types A, B, and E of Clostridium botulinum strains was performed with the use of the multiplex PCR method. Positive samples were also subjected to quantitative analysis with the use of Clostridium botulinum Isolation Agar Base (CBAB. The prevalence analysis showed 22 (21.6% samples contaminated with C. botulinum spores. The major serotype detected was botulin neurotoxin type A – 16 (72.7% whereas type B was found in 3 (13.6% honey samples and type E also only in 3 (13.6% honey samples. Dual-toxin-producing strains were noted. The average quantity of spores in PCR - C. botulinum positive samples was 190 in 1 gram of honey.

  13. 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. PMID:26887243

  14. PROPERTIES OF NEW STRAINS CHEMOLITHOTROPHIC BACTERIA ISOLATED FROM INDUSTRIAL SUBSTRATES

    Directory of Open Access Journals (Sweden)

    I. A.

    2015-12-01

    Full Text Available The purpose of the research was determination of strains Acidithiobacillus ferrooxidans MFLv37 and Acidithiobacillus ferrooxidans MFLad27, isolated from aboriginal consortium of coal beneficiation dumps and fly ash from coal combustion, resistance to heavy metals, forming part of these waste, as well as adaption ability of the strains to new substrates. New strains increased resistance to heavy metal ions as compared to A. ferrooxidans standard and collection strains is found; minimal inhibitory concentrations of heavy and toxic metals are determined; a number of metals that have negative impact on growth of isolated cultures are identified. It is shown that the minimal metals concentrations, at which strains growth still happens, are several times higher than their concentrations in technogenic waste. It has been established that isolated strains differed in their ability to adapt, as well as in growth rate and substrates oxidation. This is due to the specific conditions of microbiocenoses formation in making and further storage of rock dumps and fly ash, whereof the appropriate strains are isolated. The investigations indicate the necessity in directional selection of strains that are resistant to the toxic compounds and are able to oxidize various mineral substrates, as well as in their adaptation to new substrates for the extraction of heavy metals.

  15. 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.

  16. Botulinum Toxin

    Science.gov (United States)

    2009-01-01

    botulinum spores, which germinate and multiply into bacteria to produce neurotoxin in the food. Commonly contaminated foods include improperly preserved ... naturally occurring BoNT intoxication, it most recently occurred in Maryland as a result of a construction worker receiving a contaminated, compound...consumption and handling of meat products. Thus German physician lustinus Kerner described what he termed " sausage poisoning" (Erbguth, 2004). It was later

  17. Genome Sequence of Borrelia garinii Strain SZ, Isolated in China

    OpenAIRE

    Wu, Qiong; Liu, Zhijie; Li, Youquan; Guan, Guiquan; Niu, Qingli; Chen, Ze; Luo, Jianxun; Yin, Hong

    2014-01-01

    We announce the genome sequence of Borrelia garinii strain SZ, isolated from Dermacentor ticks collected in northeastern China. B. garinii strain SZ carries numerous plasmids, both 10 circular and 9 linear plasmids. The 902,487-bp linear chromosome (28.2% GC content) contains 820 open reading frames, 33 tRNAs, and 4 complete rRNAs. The plasmid cp32-10 contains one clustered regularly interspaced short palindromic repeat (CRISPR) with four repeats.

  18. Isolation of five Rubrobacter strains from biodeteriorated monuments

    Science.gov (United States)

    Laiz, L.; Miller, A. Z.; Jurado, V.; Akatova, E.; Sanchez-Moral, S.; Gonzalez, J. M.; Dionísio, A.; Macedo, M. F.; Saiz-Jimenez, C.

    2009-01-01

    In the last few years, the microbial colonisation of mural paintings in ancient monuments has been attracting the attention of microbiologists and conservators. The genus Rubrobacter is commonly found in biodeteriorated monuments, where it has been reported to cause rosy discolouration. However, to date, only three species of this genus have been isolated, all from thermophilic environments. In this paper, we studied three monuments: the Servilia and Postumio tombs in the Roman Necropolis of Carmona (Spain), and Vilar de Frades church (Portugal), in search of Rubrobacter strains. In all cases, biodeterioration and the formation of efflorescences were observed, and five Rubrobacter strains were isolated. These isolates showed different physiology and migration in denaturing gradient gel electrophoresis, suggesting they might represent new species within this genus. The isolates reproduced some biodeterioration processes in the laboratory and revealed their biomediation in crystal formation.

  19. Tannic acid degradation by Klebsiella strains isolated from goat feces

    Directory of Open Access Journals (Sweden)

    Arezoo Tahmourespour

    2016-03-01

    Full Text Available Background and Objectives: Tannins are toxic polyphenols that either bind and precipitate or condense proteins. The high tannin content of some plants is the preliminary limitation of using them as a ruminant feed. So, the aim of this study was the isolation and characterization of tannic acid degrading bacterial strains from goat feces before and after feeding on Pis- tachio-Soft Hulls as tannin rich diet (TRD.Materials and Methods: Bacterial strains capable of utilizing tannic acid as sole carbon and energy source were isolated and characterized from goat feces before and after feeding on TRD. Tannase activity, maximum tolerable concentration and biodegradation potential were assessed.Results: Four tannase positive isolates were identified as Klebsiella pneumoniae. Isolated strains showed the maximum tolerable concentration of 64g/L of tannin. The tannic acid degradation percentage at a concentration of 15.0 g/L reached a maximum of 68% after 24 h incubation, and more than 98% after 72 h incubation. The pH of the medium also decreased along with tannic acid utilization.Conclusions: It is obvious that TRD induced adaptive responses. Thus, while the bacteria were able to degrade and detoxify the tannic acids, they had to adapt in the presence of high concentrations of tannic acid. So, these isolates have an amazing potential for application in bioremediation, waste water treatment, also reduction of tannins antinutritional effects in animal feeds.Keywords: Biodegradation; Goat feces; Klebsiella strains; Tannic acid

  20. Historical and current perspectives on Clostridium botulinum diversity.

    Science.gov (United States)

    Smith, Theresa J; Hill, Karen K; Raphael, Brian H

    2015-05-01

    For nearly one hundred years, researchers have attempted to categorize botulinum neurotoxin-producing clostridia and the toxins that they produce according to biochemical characterizations, serological comparisons, and genetic analyses. Throughout this period the bacteria and their toxins have defied such attempts at categorization. Below is a description of both historic and current Clostridium botulinum strain and neurotoxin information that illustrates how each new finding has significantly added to the knowledge of the botulinum neurotoxin-containing clostridia and their diversity.

  1. Phylogenetic analysis of Escherichia coli strains isolated from human samples

    Directory of Open Access Journals (Sweden)

    Abdollah Derakhshandeh

    2013-12-01

    Full Text Available Escherichia coli (E. coli is a normal inhabitant of the gastrointestinal tract of vertebrates, including humans. Phylogenetic analysis has shown that E. coli is composed of four main phylogenetic groups (A, B1, B2 and D. Group A and B1 are generally associated with commensals, whereas group B2 is associated with extra-intestinal pathotypes. Most enteropathogenic isolates, however, are assigned to group D. In the present study, a total of 102 E. coli strains, isolated from human samples, were used. Phylogenetic grouping was done based on the Clermont triplex PCR method using primers targeted at three genetic markers, chuA, yjaA and TspE4.C2. Group A contained the majority of the collected isolates (69 isolates, 67.64%, followed by group B2 (18 isolates, 17.64% and D (15 isolates, 14.7% and no strains were found to belong to group B1. The distribution of phylogenetic groups in our study suggests that although the majority of strains were commensals, the prevalence of enteropathogenic and extra-intestinal pathotypes was noteworthy. Therefore, the role of E. coli in human infections including diarrhea, urinary tract infections and meningitis should be considered.

  2. 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.

  3. 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.

  4. 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.

  5. Genetic Characterization of Four Strains Borrelia Burgdorferi Isolated in China

    Institute of Scientific and Technical Information of China (English)

    曾霞; 王树声; 张涛; 毕胜利; 周永东

    2004-01-01

    To study the genetic characterization of four strains of Borrelia burgdorferi isolated in China. PCR technique was used to amplify the 5S-23S rRNA intergenic spacer DNA from the whole cellular DNA of isolated GXLD-4, 9, 18 and Chang 14, and then the amplified products were cloned into plasmid pGEM-T Easy and sequenced. It was found that the 5S-23S rRNA intergenic spacer DNA of the four isolates was 242 bp, revealing the nucleotide sequence identity of more than 99%. The four isolates had higher sequence identify with Borrelia valaisiana than with other genetic groups. These four isolates most likely belong to Borrelia valaisiana genomic group.

  6. Quality of life and social isolation in Greek adolescents with primary focal hyperhidrosis treated with botulinum toxin type A: a case series.

    Science.gov (United States)

    Kouris, Anargyros; Armyra, Kalliopi; Stefanaki, Christina; Christodoulou, Christos; Karimali, Polixeni; Kontochristopoulos, George

    2015-01-01

    Primary hyperhidrosis, although extensively documented in adults, typically has onset that dates back to early childhood. It is an unpleasant and socially disabling problem for the affected child, but little attention has been paid to the disease in adolescents. The objective of the current study was to evaluate the effectiveness of botulinum toxin type A (BTXA) in adolescents with primary palmar and axillary hyperhidrosis and to determine its effect on quality of life and social isolation. Thirty-five individuals (17 girls, 18 boys) with moderate to severe palmar and axillary hyperhidrosis were treated with BTXA (onabotulinum). Patients were examined at baseline and 6 months after treatment. The Hyperhidrosis Disease Severity Scale (HDSS) was used to evaluate disease severity and the Children's Dermatology Life Quality Index (CDLQI) was used to assess quality of life. The University of California at Los Angeles loneliness scale (UCLA version 3) was used to assess personal perception of loneliness and social isolation. The median age of the participants was 14 years, and 48.6% were female. Twenty-one had palmar hyperhidrosis, and 14 had axillary hyperhidrosis. Total CDLQI and social isolation scores decreased significantly after treatment with BTXA (both p hyperhidrosis. No statistically significant difference was documented for CDLQI and UCLA scores between boys and girls. Treatment of hyperhidrosis with BTXA resulted in improvement in quality of life, social skills, and activities.

  7. Antimicrobial resistance in Enterococcus strains isolated from healthy domestic dogs.

    Science.gov (United States)

    Bertelloni, Fabrizio; Salvadori, Claudia; Lotti, Giulia; Cerri, Domenico; Ebani, Valentina Virginia

    2016-12-15

    Enterococci are opportunistic bacteria that cause severe infections in animals and humans, capable to acquire, express, and transfer antimicrobial resistance. Susceptibility to 21 antimicrobial agents was tested by the disk diffusion method in 222 Enterococcus spp. strains isolated from the fecal samples of 287 healthy domestic dogs. Vancomycin and ampicillin minimum inhibitory concentrations (MICs) and high-level aminoglycoside resistance (HLAR) tests were also performed. Isolates showed resistance mainly to streptomycin (88.7%), neomycin (80.6%), and tetracycline (69.4%). Forty-two (18.9%) isolates showed an HLAR to streptomycin and 15 (6.7%) to gentamicin. Vancomycin and ampicillin MIC values showed 1 and 18 resistant strains, respectively. One hundred and thirty-six (61.2%) strains were classified as multidrug resistant and six (2.7%) strains as possibly extensively drug-resistant bacteria. Enterococcus faecium and Enterococcus faecalis were the most prevalent antimicrobial resistant species. Companion animals, which often live in close contact with their owners and share the same environment, represent a serious source of enterococci resistant to several antibiotics; for this reason, they may be a hazard for public health by providing a conduit for the entrance of resistance genes into the community.

  8. The use of PCR applications for assessment of Clostridium botulinum presence in the culture environment and fish samples in the conditions of open system production from Romanian fisheries

    Directory of Open Access Journals (Sweden)

    Tamara Mihociu

    2010-12-01

    Full Text Available A rapid expansion in world aquaculture has taken place in recent years due to its highestpotential to answer to the consumers demands on healthy food products. The application of biosecurityin aquaculture production systems and the safety of fish and fish products, in the context of climatechanges, requires investigations of the production systems and solutions for processing andpreservation technologies of fish products. PCR methods will be developed using the procedure ofgenes fragments amplification for Clostridium botulinum van Ermengem, 1896 species producingtoxins A, B, E and F from isolates of C. botulinum using extract broth TGPY (culture medium used forproteolytic strains and TPGYT (culture medium used for non-proteolytic strains of C. botulinum. Theresults will contribute to the integral development of food safety management and the quality of fishon the fish chain focused on the Cyprinidae for humans consumption.

  9. Analysis of the neurotoxin complex genes in Clostridium botulinum A1-A4 and B1 strains: BoNT/A3, /Ba4 and /B1 clusters are located within plasmids.

    Directory of Open Access Journals (Sweden)

    Theresa J Smith

    Full Text Available BACKGROUND: Clostridium botulinum and related clostridial species express extremely potent neurotoxins known as botulinum neurotoxins (BoNTs that cause long-lasting, potentially fatal intoxications in humans and other mammals. The amino acid variation within the BoNT is used to categorize the species into seven immunologically distinct BoNT serotypes (A-G which are further divided into subtypes. The BoNTs are located within two generally conserved gene arrangements known as botulinum progenitor complexes which encode toxin-associated proteins involved in toxin stability and expression. METHODOLOGY/PRINCIPAL FINDINGS: Because serotype A and B strains are responsible for the vast majority of human botulism cases worldwide, the location, arrangement and sequences of genes from eight different toxin complexes representing four different BoNT/A subtypes (BoNT/A1-Ba4 and one BoNT/B1 strain were examined. The bivalent Ba4 strain contained both the BoNT/A4 and BoNT/bvB toxin clusters. The arrangements of the BoNT/A3 and BoNT/A4 subtypes differed from the BoNT/A1 strains and were similar to those of BoNT/A2. However, unlike the BoNT/A2 subtype, the toxin complex genes of BoNT/A3 and BoNT/A4 were found within large plasmids and not within the chromosome. In the Ba4 strain, both BoNT toxin clusters (A4 and bivalent B were located within the same 270 kb plasmid, separated by 97 kb. Complete genomic sequencing of the BoNT/B1 strain also revealed that its toxin complex genes were located within a 149 kb plasmid and the BoNT/A3 complex is within a 267 kb plasmid. CONCLUSIONS/SIGNIFICANCE: Despite their size differences and the BoNT genes they contain, the three plasmids containing these toxin cluster genes share significant sequence identity. The presence of partial insertion sequence (IS elements, evidence of recombination/gene duplication events, and the discovery of the BoNT/A3, BoNT/Ba4 and BoNT/B1 toxin complex genes within plasmids illustrate the

  10. Isolation and characterization of organic-sulfur degradation bacterial strain

    Institute of Scientific and Technical Information of China (English)

    YANG Yu; DIAO Meng-xue; SHI Wu-yang; LI Li; DAI Qin-yun; QIU Guan-zhou

    2007-01-01

    A bacterial strain that was capable of degrading organic sulfur (dibenzothiophene) was isolated by enrichment techniques from the petroleum-contaminated soil collected from Zhongyuan Oil Field. The strain is named ZYX and is gram-positive.This strain undergoes bacilus-coccus morphological change, and forms yellow-pigment glossy circular colonies with 1.5 mm in diameter on average after 2 d incubation on Luria-Bertani(LB) plates. The full-length of 16S rDNA sequence of strain ZYX was determined and analyzed. Strain ZYX is found most relative with the genus of Arthrobacter. The similarity values between ZYX and Arthrobacter sp. P2 is 99.53%. The main morphological, biochemical and physiological features of strain ZYX accord with those of Arthrobacter. It is found that the optimal initial pH for growth is about 7.0, and the optimal concentration of dibenzothiophene(DBT)for growth is 0.10 g/L. Additionally, the results show that the best carbon source and nitrogen source are glycerol and glutamine,respectively.

  11. Properties of an Arcanobacterium haemolyticum strain isolated from a donkey.

    Science.gov (United States)

    Sammra, Osama; Balbutskaya, Anna; Nagib, Samy; Alber, Jörg; Lämmler, Christoph; Abdulmawjood, Amir; Timke, Markus; Kostrzewa, Markus; Prenger-Berninghoff, Ellen

    2014-01-01

    The present study was designed to characterize phenotypically and genotypically an Arcanobacterium haemolyticum strain (A. haemolyticum P646) isolated from a purulent nasal discharge of a donkey. A. haemolyticum P646 showed, compared to sheep blood, an enhanced hemolytic reaction on rabbit blood agar, a synergistic CAMP-like reaction with Streptococcus agalactiae and Rhodococcus equi as indicator strains, a reverse CAMP reaction in the zone of Staphylococcus aureus beta-hemolysin and the typical biochemical properties of this species. The species identity could be confirmed by MALDI-TOF MS analysis, by sequencing the 16S rDNA and glyceraldehyde-3-phosphate dehydrogenase encoding gene gap and by amplification of A. haemolyticum specific parts of 16S-23S rDNA intergenic spacer region and 23S rDNA. A. haemolyticum P646 and the reference strain A. haemolyticum DSM 20595 were further characterized by amplification of the putative virulence genes encoding arcanolysin, phospholipase D, hemolysin A, CAMP factor family protein, a collagen binding protein and two neuraminidases which were present for A. haemolyticum DSM 20595. A. haemolyticum P646 showed a comparable gene spectrum but was negative for the genes encoding collagen binding protein and neuraminidase H. To our knowledge, the present study is the first phenotypic and genotypic characterization of an A. haemolyticum strain isolated from a donkey.

  12. Evaluation of Candida strains isolated from clinical specimens

    Directory of Open Access Journals (Sweden)

    Hafize Sav

    2014-01-01

    [¤]METHODS[|]January 2011 to June 2012, Candida strains were isolated from 3905 clinical specimen. In identification of Candida species that were isolated, germ tube test, growth in Cornmeal-Tween 80 agar and formation of clamydospore, presence of pseudohyphae, carbonhytrate fermentation and assimilation tests, and the test of nitrate were studied.[¤]RESULTS[|]Finally 1122 Candida strains were isolated from 3905 various clinical specimens. The distribution of clinical specimens were as fallows: 556 from bronchoalveolar lavage (49.6%, 271 from sputum (24.2%, 114 from blood (10.2%, 51 vaginal swabs (4.6%, 50 from urine (4.4%, 30 from tissue (2.6%, 22 from endotracheal tracheal aspirate (ETA (1.9%, nine from pleural mai (0.80%, six from peritoneal fluid (0.53%, four from gastric fluid(0.35%,three from stool(0.28%,two from abscess (0.18%,three from nail (0.26%, one from cerebrospinal fluid (0.10%. From these clinical samples 848 C. albicans (75.6%, 143 C. glabrata (12.8%, 40 C. parapsilosis, (3.57%, 33 C. krusei (2.94%, 33 C. kefyr (2.94%, 19 C. tropicalis (1.7% were isolated. Other strains were identified as C. lusitania, C. lipolytica, C. norvegensis, C. pelliculosa ve C. zeylanoides.[¤]CONCLUSION[|] It was concluded that C.albicans has still been the most frequent species among Candida isolates of in our hospital; however, the incidence of non-albicans species have increased.[¤

  13. 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.

  14. Isolation and identification of the thermophilic alkaline desulphuricant strain

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A desulfurization strain that belongs to the thermophilic alkaline desulphuricant is designated as strain GDJ-3 and isolated from Inner Mongolia, China. The colony of the strain shows tiny, yellow, or white-yellow, and it becomes henna with the protracting of cultivated time. The cells are bacilliform (0.3 -0.6 × 1.0-1.2 μm), motive, and Gram negative. The strain GDJ-3 is able to utilize respectively the thiosulphate, sulfate, sulfite, or sulfide as sulfur source, utilize the carbon dioxide as the carbon source, and utilize the ammonium or nitrate as the nitrogen source. According to GenBank data, 16s RNA results of GDJ-3 are in good agreement with Alpha proteobacterrium sp. (97%) and Ochrobactrum sp. (98%). For GDJ-3, the optimum growth temperature is at 45℃, the optimum pH is at 8.5-8.8, and the optimum rocking speed of sorting table is at 150 r/min. Under the optimum culture condition, the cells of the strain can live for about 18 h. In the desulfurization solution, which is prepared according to the composition of DDS solution, the objectionable constituents of sodium thiosulphate and sodium sulfide were added factitiously, and the bacterial cell concentration was set at 107/mL. After the regeneration of the above desulfurization solution by the strain cells, the concentration of sodium thiosulphate was decreased by 14.75 g/L (percentage loss of content 13.21%), the concentration of sodium sulfide was decreased by 0.76 g/L (percentage loss of content 87.36%) in the desulfurization solution in 9.5 hours, and sulfur appeared. Maybe, this kind of strain can be used as the regeneration’s bacterial source of DDS solution.

  15. 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.

  16. 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.

  17. 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.

  18. [Intoxication of botulinum toxin].

    Science.gov (United States)

    Chudzicka, Aleksandra

    2015-09-01

    Botulinum toxin is an egzotoxin produced by Gram positive bacteria Clostridium botulinum. It is among the most potent toxins known. The 3 main clinical presentations of botulism are as follows: foodborne botulism, infant botulism and wound botulism. The main symptom of intoxication is flat muscles paralysis. The treatment is supportive care and administration of antitoxin. In prevention the correct preparing of canned food is most important. Botulinum toxin is accepted as a biological weapon.

  19. 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

  20. Draft Genome Sequences of Two Aspergillus fumigatus Strains, Isolated from the International Space Station.

    Science.gov (United States)

    Singh, Nitin Kumar; Blachowicz, Adriana; Checinska, Aleksandra; Wang, Clay; Venkateswaran, Kasthuri

    2016-07-14

    Draft genome sequences of Aspergillus fumigatus strains (ISSFT-021 and IF1SW-F4), opportunistic pathogens isolated from the International Space Station (ISS), were assembled to facilitate investigations of the nature of the virulence characteristics of the ISS strains to other clinical strains isolated on Earth.

  1. Genome Sequence of a Helicobacter pylori Strain Isolated from a Mexican Patient with Intestinal Gastric Cancer

    Science.gov (United States)

    Larios-Serrato, Violeta; Olguín-Ruiz, Gabriela Edith; Sánchez-Vallejo, Carlos Javier; Torres-López, Roberto Carlos; Avilés-Jiménez, Francisco; Camorlinga-Ponce, Margarita

    2014-01-01

    Helicobacter pylori strains are the major risk factor for gastric cancer. Strains vary in their content of disease-associated genes, so genome-wide analysis of cancer-isolated strains will help elucidate their pathogenesis and genetic diversity. We present the draft genome sequence of H. pylori isolated from a Mexican patient with intestinal gastric cancer. PMID:24459275

  2. 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.

  3. 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

    Campylobacter is one of the leading causes of food-borne gastroenteritis and has a high prevalence in poultry. Campylobacter jejuni subsp. jejuni 327 is a subspecies of the genus Campylobacter of the family Campylobacteraceae in the phylum Proteobacteria. The microaerophilic, spiral shaped, catal...... 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)....

  4. Isolation and Characterization of Heterotrophic Nitrifying Strain W1

    Institute of Scientific and Technical Information of China (English)

    吕永康; 王荀; 刘博凯; 刘玉香; 杨晓华

    2012-01-01

    In a high concentration substrate medium, a heterotrophic bacterium with high removal efficiency of ammonium, named W1, was isolated from activated sludge of coking wastewater treatment facility. The bacterium was Gram-negative, rod-shaped, and identified preliminarily as Alcaligenes sp. according to its morphological and physiological properties and its 16S rRNA gene sequence analysis. In the high concentration ammonium medium (400 mg·L 1 4 NH -N), the effects of C source, N source, C/N ratio and initial pH of medium on ammonium removal were investigated in order to determine the optimal condition for strain W1. The maximum ammonium removal was around 95% in 4 days in an improved medium. The production of N 2 gas was examined in a closed system that was full of pure oxygen at the beginning. N 2 gas was detected in the system after 4 days of cultivation, which further testified that strain W1 has heterotrophic nitrification and aerobic denitrification abilities simultaneously.

  5. Decolorization of the textile dyes by newly isolated bacterial strains.

    Science.gov (United States)

    Chen, Kuo-Cheng; Wu, Jane-Yii; Liou, Dar-Jen; Hwang, Sz-Chwun John

    2003-02-27

    Six bacterial strains with the capability of degrading textile dyes were isolated from sludge samples and mud lakes. Aeromonas hydrophila was selected and identified because it exhibited the greatest color removal from various dyes. Although A. hydrophila displayed good growth in aerobic or agitation culture (AGI culture), color removal was the best in anoxic or anaerobic culture (ANA culture). For color removal, the most suitable pH and temperature were pH 5.5-10.0 and 20-35 degrees C under anoxic culture (ANO culture). More than 90% of RED RBN was reduced in color within 8 days at a dye concentration of 3,000 mg l(-1). This strain could also decolorize the media containing a mixture of dyes within 2 days of incubation. Nitrogen sources such as yeast extract or peptone could enhance strongly the decolorization efficiency. In contrast to a nitrogen source, glucose inhibited decolorization activity because the consumed glucose was converted to organic acids that might decrease the pH of the culture medium, thus inhibiting the cell growth and decolorization activity. Decolorization appeared to proceed primarily by biological degradation.

  6. Botulinum Toxin (Botox) for Facial Wrinkles

    Science.gov (United States)

    ... Stories Español Eye Health / Eye Health A-Z Botulinum Toxin (Botox) for Facial Wrinkles Sections Botulinum Toxin (Botox) ... Facial Wrinkles How Does Botulinum Toxin (Botox) Work? Botulinum Toxin (Botox) for Facial Wrinkles Written by: Kierstan Boyd ...

  7. Observations on the distribution and ecology of Clostridium botulinum type E in Alaska.

    Science.gov (United States)

    Miller, L G

    1975-06-01

    Environmental samples collected along the coastline and from the interior of Alaska were examined for the presence of Clostridium botulinum. Clostridium botulinum type E was detected in soils from 5 of 12 beaches; in 7 of 115 non-coastal soil samples; in sediments from six of eight locales; in gills of salmon from two fishing areas; and in the feces of 1 of 44 colonic samples from marine mammals. The basic biochemical characteristics of the isolates were determined. Tube tests for demonstrating gelatin liquefaction proved insensitive with these strains, whereas a plate test detected gelatinase in all isolates. The presence of multiple nidi and the continual discharge of organic materials into the environment may contribute to the perpetuation of botulinum spores by which foods prepared form marine animals become contaminated. An emphasis should be placed upon the need for measures to reduce environmental contamination, to reduce contamination during food preparation, and to alert continually the population of the hazard wherever botulism is endemic.

  8. Botulinum Neurotoxins

    Science.gov (United States)

    2012-03-28

    200,000 people were infected with an antibiotic -resistant strain of Salmonella caused by an inadvertent contamination at a single northern Illinois dairy...Fenecia et al., 1999). The disease has also been correlated with alteration of protective endogenous microflora by broad-spectrum antibiotics after... equine antitoxin, and discharged after 9 days. While not a naturally occurring form of the disease, inhalational botulism has implications as a

  9. 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.

  10. 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.

  11. 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.

  12. Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry.

    Science.gov (United States)

    Stępień-Pyśniak, Dagmara; Marek, Agnieszka; Banach, Tomasz; Adaszek, Łukasz; Pyzik, Ewelina; Wilczyński, Jarosław; Winiarczyk, Stanisław

    2016-06-01

    The aim of this study was to evaluate the frequency of occurrence of bacteria of the genus Enterococcus in poultry, to identify them by means of matrixassisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF MS), and to analyse the antimicrobial susceptibility of the isolated strains to the drugs most frequently used in poultry. The material for the bacteriological tests was obtained mainly from the heart (97%) of the birds investigated. Of a total of 2,970 samples tested, 911 (30.7%) tested positive for Enterococcus spp. Enterococci were detected in broilers (88.1%), laying hens (5.3%), turkeys (3.9%), breeding hens (2.2%), and geese (0.4%). The most commonly identified species were Enterococcus (E.) faecalis (74.7%), E. faecium (10.1%), E. gallinarum (5.5%), E. hirae (4.6%), and E. cecorum (4.1%). The most frequent resistance properties were resistance to sulphamethoxazole/trimethoprim (88%), tylosin (71.4%), enrofloxacin (69.4%), doxycycline (67.3%), and lincomycin/spectinomycin (56.1%). Only one vancomycin-resistant Enterococcus, E. cecorum from a broiler, was found.

  13. Draft Genome of the Multidrug-Resistant Acinetobacter baumannii Strain A155 Clinical Isolate.

    Science.gov (United States)

    Arivett, Brock A; Fiester, Steven E; Ream, David C; Centrón, Daniela; Ramírez, Maria S; Tolmasky, Marcelo E; Actis, Luis A

    2015-03-26

    Acinetobacter baumannii is a bacterial pathogen with serious implications on human health, due to increasing reports of multidrug-resistant strains isolated from patients. Total DNA from the multidrug-resistant A. baumannii strain A155 clinical isolate was sequenced to greater than 65× coverage, providing high-quality contig assemblies.

  14. CHARACTERIZATION OF A NEW BACILLUS-STEAROTHERMOPHILUS ISOLATE - A HIGHLY THERMOSTABLE ALPHA-AMYLASE-PRODUCING STRAIN

    NARCIS (Netherlands)

    WIND, RD; BUITELAAR, RM; EGGINK, G; HUIZING, HJ; DIJKHUIZEN, L

    1994-01-01

    A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known alpha-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable alpha-amylase. The half-time of inactivation of this alpha-amylas

  15. Characterization of a new Bacillus stearothermophilus isolate : a highly thermostable α-amylase-producing strain

    NARCIS (Netherlands)

    Wind, R.D.; Buitelaar, R.M.; Eggink, G.; Huizing, H.J.; Dijkhuizen, L.

    1994-01-01

    A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known α-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable α-amylase. The half-time of inactivation of this α-amylase was 5.1 h

  16. Primary Isolation Strain Determines Both Phage Type and Receptors Recognised by Campylobacter jejuni Bacteriophages

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina;

    2015-01-01

    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......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...... 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...

  17. Oligomycins A and C, major secondary metabolites isolated from the newly isolated strain Streptomyces diastaticus.

    Science.gov (United States)

    Yang, P W; Li, M G; Zhao, J Y; Zhu, M Z; Shang, H; Li, J R; Cui, X L; Huang, R; Wen, M L

    2010-01-01

    During the screening program for fungicides, one actinomycete strain ECO 00047 was isolated with the potential activity against fungus. According to the morphology and analysis of the nucleotide sequence of the 16S rRNA gene (1500 bp) this isolate was identified as Streptomyces diastaticus. The active compounds were separated by silica gel column chromatography, Sephadex LH-20 gel filtration and then purified by flash chromatography on C18 (20-45 microm). The chemical structure of the bioactive compounds I and II were elucidated, based on the spectroscopic data of MS, IR, UV, 1H-NMR, 13C-NMR and X-ray single crystal diffraction analysis. Compounds I and II were identical with oligomycins A and C, the macrolide antibiotics which have been known to be produced by Streptomyces diastatochromogenes, S. libani and S. avermitilis. The two compounds exhibited a strong activity against Aspergillus niger, Alternaria alternata, Botrytis cinerea and Phytophthora capsici but no activity toward bacteria. Although the two above antibiotics were known, their isolation has so far not been reported from S. diastaticus.

  18. Genome sequences of six Pseudoalteromonas strains isolated from Arctic sea ice.

    Science.gov (United States)

    Bian, Fei; Xie, Bin-Bin; Qin, Qi-Long; Shu, Yan-Li; Zhang, Xi-Ying; Yu, Yong; Chen, Bo; Chen, Xiu-Lan; Zhou, Bai-Cheng; Zhang, Yu-Zhong

    2012-02-01

    Yu et al. (Polar Biol. 32:1539-1547, 2009) isolated 199 Pseudoalteromonas strains from Arctic sea ice. We sequenced the genomes of six of these strains, which are affiliated to different Pseudoalteromonas species based on 16S rRNA gene sequences, facilitating the study of physiology and adaptation of Arctic sea ice Pseudoalteromonas strains.

  19. 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.

  20. Genome Sequence of Gluconacetobacter sp. Strain SXCC-1, Isolated from Chinese Vinegar Fermentation Starter▿

    OpenAIRE

    Du, Xin-jun; Jia, Shi-Ru; Yang, Yue; Wang, Shuo

    2011-01-01

    Gluconacetobacter strains are prominent bacteria during traditional vinegar fermentation. Here, we report a draft genome sequence of Gluconacetobacter sp. strain SXCC-1. This strain was isolated from a fermentation starter (Daqu) used for commercial production of Shanxi vinegar, the best-known vinegar of China.

  1. Draft Genome Sequence of Kluyveromyces marxianus Strain DMB1, Isolated from Sugarcane Bagasse Hydrolysate.

    Science.gov (United States)

    Suzuki, Toshihiro; Hoshino, Tamotsu; Matsushika, Akinori

    2014-07-24

    We determined the genome sequence of a thermotolerant yeast, Kluyveromyces marxianus strain DMB1, isolated from sugarcane bagasse hydrolysate, and the sequence provides further insights into the genomic differences between this strain and other reported K. marxianus strains. The genome described here is composed of 11,165,408 bases and has 4,943 protein-coding genes.

  2. Efficient screening of environmental isolates for Saccharomyces cerevisiae strains that are suitable for brewing.

    Science.gov (United States)

    Fujihara, Hidehiko; Hino, Mika; Takashita, Hideharu; Kajiwara, Yasuhiro; Okamoto, Keiko; Furukawa, Kensuke

    2014-01-01

    We developed an efficient screening method for Saccharomyces cerevisiae strains from environmental isolates. MultiPlex PCR was performed targeting four brewing S. cerevisiae genes (SSU1, AWA1, BIO6, and FLO1). At least three genes among the four were amplified from all S. cerevisiae strains. The use of this method allowed us to successfully obtain S. cerevisiae strains.

  3. [Plague in Algeria: about five strains of Yersinia pestis isolated during the outbreak of June 2003].

    Science.gov (United States)

    Lounici, M; Lazri, M; Rahal, K

    2005-02-01

    In this study, we isolated and identified five strains of Yersinia pestis during an epidemic occurred in west of Algeria in June 2003. The bacteriological identification was confirmed by bacteriophage susceptibility. All these strains belonged to the biovar Orientalis (they did not ferment glycerol but did reduce nitrate to nitrite) which caused the current pandemic. The in vitro activities of antimicrobial agents used to treat plague and recommended for prophylaxis, showed that they are active against all strains. The comparison of these strains by plasmid profile analyse demonstrated that all isolates had three plasmids: 110, 70 and 9.5 kb, which are present in Y. pestis strains.

  4. Immunological comparison of scrapie-associated fibrils isolated from animals infected with four different scrapie strains.

    OpenAIRE

    Kascsak, R J; Rubenstein, R; Merz, P A; Carp, R I; Robakis, N. K.; Wisniewski, H M; Diringer, H

    1986-01-01

    Scrapie-associated fibrils (SAFs) are abnormal filamentous structures that are uniquely associated with unconventional slow virus diseases. The antigenic relationships of SAFs from animals infected with four biologically distinct scrapie strains were investigated by using antisera raised to purified SAF proteins. Rabbit antisera were raised to SAFs isolated from mice infected with the ME7 scrapie strain and to SAFs isolated from hamsters infected with the 263K scrapie strain. A strong antigen...

  5. 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

  6. Distribution of uropathogenic virulence factors among Escherichia coli strains isolated from dogs and cats.

    Science.gov (United States)

    Yuri, K; Nakata, K; Katae, H; Yamamoto, S; Hasegawa, A

    1998-03-01

    A variety of virulence factors (VFs) such as type 1 fimbriae, pilus associated with pyelonephritis, S fimbriae, afimbrial adhesin, alpha-hemolysin, aerobactin and cytotoxic necrotizing factor 1 are associated with uropathogenic Escherichia coli. In this study, 80 uropathogenic E. coli strains in 50 dogs and 30 cats suffering from UTI. In addition, 60 E. coli strains were isolated from fecal samples from 30 each of healthy dogs and cats. The distribution of VFs of uropathogenic E. coli strains isolated from dogs and cats suffering from urinary tract infections (UTI) were examined by the colony hybridization test with seven DNA probes specific for VFs, and the results were compared with those obtained in the studies on strains from humans with UTI. In uropathogenic E. coli strains isolated from dogs and cats suffering from UTI, VFs were detected as frequently as in the strains isolated from humans with UTI. Although less frequently, genes encoding these VFs especially pap, sfa, hly, and cnf 1 genes were also associated with E. coli strains isolated from feces of healthy cats, in contrast to the distribution pattern of uropathogenic E. coli observed in humans. Furthermore, all VFs except pil were significantly more frequently detected in strains isolated from urine of animals with cystitis than in those isolated from feces of healthy humans. These results indicate that VFs of E. coli contribute to the pathogenesis of UTI in dogs and cats.

  7. Identification and biochemical characterization of Leishmania strains isolated in Peru, Mexico, and Spain.

    Science.gov (United States)

    Rodríguez-González, Isabel; Marín, Clotilde; Vargas, Franklin; Córdova, Ofelia; Barrera, Mario; Gutiérrez-Sánchez, Ramón; Alunda, Jose María; Sánchez-Moreno, Manuel

    2006-01-01

    Eight Leishmania promastigotes were isolated from different geographical areas: three (LP1, LP2, and LP3) from the provincial department La Libertad and the fourth (LP4) from the department of Cajamarca (northern Peru); another three (LM1, LM2, and LM3) in the province of Campeche (Mexico); and the last (LS1) from a clinical case of a dog in Madrid (Spain). The isolates were characterized by carbohydrate cell-surface residues using agglutinations with four purified lectins, by isoenzyme analysis using different isoenzymes, by analysis of kinetoplast DNA (kDNA) restriction fragment length polymorphism using four different restriction endonucleases and by the final metabolite patterns after in vitro culture. These isolates were compared with four reference strains and typified as: Leishmania (Leishmania) donovani, two strains of L. (L.) infantum, and one species of L. (Viania) peruviana. According to our results and the statistical study, the Peruvian isolates represent three different strains: one would be L. (V.) peruviana, another the strain isolated in Cajamarca (LP4) and the third would include the three strains from the department of La Libertad (LP1, LP2, and LP3), these latter three isolates being phylogenetically closer to the reference strain L. (L.) donovani. Meanwhile, the three isolates from Mexico form a group with close phylogenetic relationships to each other. The isolate from Spain belongs to the species L. (L.) infantum. Thus, a close correlation was drawn between the identity of each strain and its geographical origin.

  8. 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...... strains, as demonstrated on a further six T. gondii reference strains [BK and GT1 (group A), NTE and 561 (group B), and NED and C56 (group C)]. The bradyzoite-specific mAb, 4.3, reacted to the GT1, NTE and 561 strains, but not to the BK, NED or C56 strains. The tachyzoite-specific mAb, 4.25, reacted....... gondii strain collection representative for a small geographic area (Denmark) was established within a short time span from a variety of animal species. Using the mAb as typing reagents to this Danish strain collection, all 36 animal and two human strains were identified as having the same reaction...

  9. Historical Perspectives and Guidelines for Botulinum Neurotoxin Subtype Nomenclature

    Science.gov (United States)

    Peck, Michael W.; Smith, Theresa J.; Anniballi, Fabrizio; Austin, John W.; Bano, Luca; Bradshaw, Marite; Cuervo, Paula; Cheng, Luisa W.; Derman, Yagmur; Dorner, Brigitte G.; Fisher, Audrey; Hill, Karen K.; Kalb, Suzanne R.; Korkeala, Hannu; Lindström, Miia; Lista, Florigio; Lúquez, Carolina; Mazuet, Christelle; Pirazzini, Marco; Popoff, Michel R.; Rossetto, Ornella; Rummel, Andreas; Sesardic, Dorothea; Singh, Bal Ram; Stringer, Sandra C.

    2017-01-01

    Botulinum neurotoxins are diverse proteins. They are currently represented by at least seven serotypes and more than 40 subtypes. New clostridial strains that produce novel neurotoxin variants are being identified with increasing frequency, which presents challenges when organizing the nomenclature surrounding these neurotoxins. Worldwide, researchers are faced with the possibility that toxins having identical sequences may be given different designations or novel toxins having unique sequences may be given the same designations on publication. In order to minimize these problems, an ad hoc committee consisting of over 20 researchers in the field of botulinum neurotoxin research was convened to discuss the clarification of the issues involved in botulinum neurotoxin nomenclature. This publication presents a historical overview of the issues and provides guidelines for botulinum neurotoxin subtype nomenclature in the future. PMID:28106761

  10. Historical Perspectives and Guidelines for Botulinum Neurotoxin Subtype Nomenclature

    Directory of Open Access Journals (Sweden)

    Michael W. Peck

    2017-01-01

    Full Text Available Botulinum neurotoxins are diverse proteins. They are currently represented by at least seven serotypes and more than 40 subtypes. New clostridial strains that produce novel neurotoxin variants are being identified with increasing frequency, which presents challenges when organizing the nomenclature surrounding these neurotoxins. Worldwide, researchers are faced with the possibility that toxins having identical sequences may be given different designations or novel toxins having unique sequences may be given the same designations on publication. In order to minimize these problems, an ad hoc committee consisting of over 20 researchers in the field of botulinum neurotoxin research was convened to discuss the clarification of the issues involved in botulinum neurotoxin nomenclature. This publication presents a historical overview of the issues and provides guidelines for botulinum neurotoxin subtype nomenclature in the future.

  11. Important role of class I heat shock genes hrcA and dnaK in the heat shock response and the response to pH and NaCl stress of group I Clostridium botulinum strain ATCC 3502.

    Science.gov (United States)

    Selby, Katja; Lindström, Miia; Somervuo, Panu; Heap, John T; Minton, Nigel P; Korkeala, Hannu

    2011-05-01

    Class I heat shock genes (HSGs) code for molecular chaperones which play a major role in the bacterial response to sudden increases of environmental temperature by assisting protein folding. Quantitative reverse transcriptase real-time PCR gene expression analysis of the food-borne pathogen Clostridium botulinum grown at 37°C showed that the class I HSGs grpE, dnaK, dnaJ, groEL, and groES and their repressor, hrcA, were expressed at constant levels in the exponential and transitional growth phases, whereas strong downregulation of all six genes was observed during stationary phase. After heat shock from 37 to 45°C, all HSGs were transiently upregulated. A mutant with insertionally inactivated hrcA expressed higher levels of class I HSGs during exponential growth than the wild type, followed by upregulation of only groES and groES after heat shock. Inactivation of hrcA or of dnaK encoding a major chaperone resulted in lower maximum growth temperatures than for the wild type and reduced growth rates under optimal conditions compared to the wild type. The dnaK mutant showed growth inhibition under all tested temperature, pH, and NaCl stress conditions. In contrast, the growth of an hrcA mutant was unaffected by mild temperature or acid stress compared to the wild-type strain, indicating that induced class I HSGs support growth under moderately nonoptimal conditions. We show that the expression of class I HSGs plays a major role for survival and growth of C. botulinum under the stressful environmental conditions that may be encountered during food processing or growth in food products, in the mammalian intestine, or in wounds.

  12. Polymorphism of Beauveria bassiana (Deuteromycota: Hyphomycetes) strains isolated from Ixodes ricinus (Acari: Ixodidae) in Moldova.

    Science.gov (United States)

    Mitina, G V; Tokarev, Y S; Movila, A A; Yli-Mattila, T

    2011-03-01

    Polymorphism of 10 Beauveria bassiana strains, isolated from Ixodes ricinus in Moldova, was evaluated using traditional (morphological and cultural properties) and molecular (RAPD patterns and ITS sequences) methods. The isolates differed greatly in morphological and cultural features, such as color, consistence, and growth rate. Four RAPD-PCR markers were used to evaluate genetic diversity of the strains. Phylogenetic neighbor-joining analysis of RAPD patterns divided strains into 3 major clades. The ITS sequences of 8 strains were identical to those of known B. bassiana strains. Two subsets (1 and 2) different by one nucleotide change were found in the ITS1 region. One strain of subset 1 was different from known B. bassiana strains by possessing 2 point mutations in the ITS region. RAPD-based clustering correlated to ITS sequence and colony morphology-based grouping of the strains.

  13. Molecular characterization of Acidithiobacillus ferrooxidans and A. thiooxidans strains isolated from mine wastes in Brazil.

    Science.gov (United States)

    Paulino, L C; Bergamo, R F; Garcia, O; de Mello, M P; Manfio, G P; Ottoboni, L M

    2001-10-01

    Nineteen strains of Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans, including 12 strains isolated from coal, copper, gold and uranium mines in Brazil, strains isolated from similar sources in other countries and the type strains of the two species were characterized together with the type strain of A. caldus by using a combination of molecular systematic methods, namely ribotyping, BOX- and ERIC-PCR and DNA-DNA hybridization assays. Data derived from the molecular fingerprinting analyses showed that the tested strains encompassed a high degree of genetic variability. Two of the Brazilian A. ferrooxidans organisms (strains SSP and PCE) isolated from acid coal mine waste and uranium mine effluent, respectively, and A. thiooxidans strain DAMS, isolated from uranium mine effluent, were the most genetically divergent organisms. The DNA-DNA hybridization data did not support the allocation of Acidithiobacillus strain SSP to the A. ferrooxidans genomic species, as it shared only just over 40% DNA relatedness with the type strain of the species. Acidithiobacillus strain SSP was not clearly related to A. ferrooxidans in the 16S rDNA tree.

  14. Characterization of a new clinical yeast species isolated from a strain collection of Tunisian hospitals

    NARCIS (Netherlands)

    Eddouzi, J.; Hofstetter, V.; Groenewald, M.; Manai, M.; Sanglard, S.

    2013-01-01

    From a collection of yeast isolates isolated from patients in Tunisian hospitals between September 2006 and July 2010, the yeast strain JEY63 (CBS 12513), isolated from a 50-year-old male that suffered from oral thrush, could not be identified to the species level using conventional methods used in

  15. Full-Genome Sequences of Seven Fatal Enterovirus 71 Strains Isolated in Shenzhen, China, in 2014

    Science.gov (United States)

    He, Ya-Qing; Meng, Jun; Xiong, Ling-Hong; Wang, Chao; Yao, Xiang-Jie; Zhang, Hai-Long; Zhang, Ren-Li

    2016-01-01

    The whole-genome sequences of seven fatal enterovirus 71 (EV71) strains, isolated in southern China, in 2014, were determined. The complete genome sequences of these strains displayed close relationships to native EV71 strains and showed 94.2% to 99.8% identity to each other. All of these strains were assigned to subgenotype C4a based on phylogenetic analysis of the VP1 gene. PMID:27125487

  16. Phenotypic and genotypic diversity of Pseudomonas aeruginosa strains isolated from hospitals in siedlce (Poland

    Directory of Open Access Journals (Sweden)

    Katarzyna Wolska

    2012-03-01

    Full Text Available A total of 62 Pseudomonas aeruginosa strains isolated from two hospitals in Siedlce (Poland were studied by repetitive element based PCR (rep-PCR using BOX primer. BOX-PCR results revealed the presence of 7 numerous genotypes and 31 unique patterns among isolates. Generally, the strains of P. aeruginosa were characterized by resistance to many antibiotics tested and by differences in serogroups and types of growth on cetrimide agar medium. However, the P. aeruginosa strains isolated from faeces showed much lower phenotypic and genotypic variations in comparison with strains obtained from other clinical specimens. It was observed that genetic techniques supported by phenotypic tests have enabled to conduct a detailed characterization of P. aeruginosa strains isolated from a particular environment at a particular time.

  17. 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.

  18. 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.

  19. Genotypic characteristics of bovine viral diarrhea virus 2 strains isolated in northern Italy.

    Science.gov (United States)

    Giangaspero, M; Harasawa, R; Zecconi, A; Luzzago, C

    2001-09-01

    Two strains of Bovine viral diarrhea virus 2 (BVDV-2) were isolated from calves in northern Italy. Variations in the 5'-untranslated region (UTR) of the genome were studied by primary structure alignment and neighbor-joining method based phylogenetic tree analyses and by palindromic nucleotide substitutions at the three variable loci in the 5'-UTR. Genetic analysis indicated their appurtenance to genovar BVDV-2a. Nucleotide sequence at the 5'-UTR of strain BS-95-II, one of the Italian isolates from healthy calves, showed 98% homology to that of the Japanese isolate OY89, a cytopathic strain derived from cattle with mucosal disease.

  20. Serologic response of vaccinated cattle to strains of Moraxella bovis isolated during epizootics of keratoconjunctivitis.

    Science.gov (United States)

    Pugh, G W; Hughes, D E; Booth, G D

    1978-01-01

    In studies to determine whether there were antigenic differences between strains (isolates) of Moraxella bovis, the sera from vaccinated calves were tested with isolates of M bovis while the calves were experiencing epizootics of infectious bovine keratoconjunctivitis (IBK). Before the epizootics of IBK, the calves were intramuscularly vaccinated with a formalin-killed autogenous M bovis bacterin. During the epizootics, the eyes were examined by cultural technique, and isolates which were obtained were categorized by catalase activity, source (diseased or nondiseased eyes), and reactivity with the various sera. The serum reactivity of the isolates was compared with that of the vaccinal strain. The vaccinal strain and 8 of the 1 5 selected isolates obtained during the 1974 epizootic were catalase negative. Seven of the 15 isolates from the 1974 epizootic and all of the selected isolates from the 1975 epizootic were catalase positive. A significantly higher (P less than 0.01) percentage of calf sera were serologically reactive with the vaccinal strain and other catalase-negative isolates (45.0%) than with catalase-positive isolates (34.8%). The results, although not definitive, suggest that there may be antigenic differences among strains of M bovis. These differences should be considered when cattle are vaccinated against IBK under natural conditions of exposure.

  1. Characterization of Salmonella Enteritidis strains isolated from poultry and farm environments in Brazil.

    Science.gov (United States)

    Campioni, F; Zoldan, M M; Falcão, J P

    2014-07-01

    Salmonella Enteritidis is a major causative agent of foodborne outbreaks worldwide. Using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and pulsed-field gel electrophoresis (PFGE), this study assessed the genetic relatedness, the pathogenic potential, and antimicrobial resistance in 60 strains isolated from chickens and the farm environment in Brazil between 2004 and 2010. The resulting concatenated dendrogram of the two methodologies distinguished the strains into two clusters. Some strains isolated from the two sources were indistinguishable. All the strains contained the 13 virulence markers investigated. Forty-four strains were resistant to nalidixic acid. Quinolone resistance presented by many strains suggests that quinolones may have been used to treat chickens. The high prevalence of virulence markers highlights the importance of poultry as vehicles of S. Enteritidis strains that have the potential to cause disease.

  2. 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. ...

  3. Identification and biocellulose production of Gluconacetobacter strains isolated from tropical fruits in Thailand

    Directory of Open Access Journals (Sweden)

    Daungjai Ochaikul

    2013-02-01

    Full Text Available Two hundred and four strains of biocellulose (BC-producing Gluconacetobacter strains were isolated from 48 rotten tropical fruits collected in Thailand. Twenty-nine representative isolates were selected from each of the 16 isolation sources and identified by morphological, physiological and biochemical characteristics and 16S rRNA gene sequence analysis. The selected 29 isolates were divided into seven subgroups within the Gluconacetobacter xylinus group of the genus Gluconacetobacter and identified as Gluconacetobacter oboediens (subgroup I, five isolates, Gluconacetobacter rhaeticus (subgroup II, one isolate, Gluconacetobacter hansenii (subgroup III, seven isolates, Gluconacetobacter swingsii (subgroup IV, two isolates and Gluconacetobacter sucrofermentans (subgroup V, two isolates. The remaining isolates were grouped into subgroups VIa (three isolates and VIb (nine isolates. All the isolates were cultured in Hestrin-Schramm (HS medium statically at 30C for 7 days to determine cellulose production capability. Of the 29 isolates, isolate PAP1 (subgroup VIb, unidentified gave the highest yield (1.15 g/L of BC. However, the BC yield increased threefold (3.5 g/L when D-glucose in HS medium was replaced by D-mannitol.

  4. Isolation and characterization of multiply antibiotic-resistant Clostridum perfringens strains from porcine feces.

    Science.gov (United States)

    Rood, J I; Maher, E A; Somers, E B; Campos, E; Duncan, C L

    1978-05-01

    Multiply antibiotic-resistant strains of Clostridium perfringens were isolated from porcine feces. Strains that were resistant to tetracycline, erythromycin, clindamycin, and lincomycin were isolated, but no penicillin- or chloramphenicol-resistant strains were obtained. Typical minimal inhibitory concentrations for resistant strains were 16 to 64 mug of tetracycline per ml, 64 to >128 mug of erythromycin per ml, >/=128 mug of lincomycin per ml, and 16 to 128 mug of clindamycin per ml. Resistance to erythromycin was always associated with resistance to lincomycin and clindamycin. Minimal inhibitory concentrations were determined for 258 strains from six farms that used antibiotics in their feeds and 240 strains from five farms that did not use antibiotics. The results show that 77.9 and 22.7% of the strains from the former farms were resistant to tetracycline and erythromycin-clindamycin-lincomycin, respectively. The comparable data from the latter farms were 25.0 and 0.8%, respectively. Agarose gel electrophoresis failed to reveal a plasmid band that was common to the resistant strains but absent in the susceptible strains. Attempts to transfer tetracycline, erythromycin, and clindamycin resistance from one strain, CW459, were not successful. Antibiotic-susceptible mutants were not isolated from this strain, despite the use of a variety of curing agents.

  5. Genotypic diversity of Lactobacillus sanfranciscensis strains isolated from French organic sourdoughs.

    Science.gov (United States)

    Lhomme, Emilie; Onno, Bernard; Chuat, Victoria; Durand, Karine; Orain, Servane; Valence, Florence; Dousset, Xavier; Jacques, Marie-Agnès

    2016-06-01

    Lactobacillus sanfranciscensis is the predominant key lactic acid bacterium in traditionally fermented sourdoughs. Despite its prevalence, sourdough and their related breads could be different regarding their physicochemical and sensorial characteristics. The intraspecific diversity of L. sanfranciscensis might explain these observations. Fifty-nine strains isolated from French sourdoughs were typed by a polyphasic approach including Multilocus Sequence Typing (MLST) and Pulsed-field Gel Electrophoresis (PFGE), in order to study their genotypic diversity. MLST scheme can be reduced from six to four gene fragments (gdh, gyrA, nox and pta) without a major loss of discrimination between strains. The genes mapA and pgmA are not good candidates for inclusion in an MLST scheme to type L. sanfranciscensis strains, as they could not be amplified for a set of 18 strains among the 59 studied. This method revealed 20 sequence types (STs). Of these, 19 STs were grouped in one clonal complex, showing a strong relatedness between these strains. PFGE using SmaI discriminated 41 pulsotypes and so distinguished isolates better than the MLST scheme. Both genotypic methods indicate a low diversity between strains isolated from the same sourdough and a higher diversity between strains isolated from different sourdoughs, suggesting an influence of baker practices and/or environmental conditions on the selection of strains. The use of these two methods targeting genetic variations gives an optimal genotypic characterization of L.sanfranciscensis strains.

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

    Science.gov (United States)

    Bernal, Milagro García; Campa-Córdova, Ángel Isidro; Saucedo, Pedro Enrique; González, Marlen Casanova; Marrero, Ricardo Medina; Mazón-Suástegui, José Manuel

    2015-01-01

    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. PMID:27047067

  7. Draft Genome Sequence of the Field Isolate Brucella melitensis Strain Bm IND1 from India.

    Science.gov (United States)

    Rao, Sashi Bhushan; Gupta, Vivek K; Kumar, Mukesh; Hegde, Nagendra R; Splitter, Gary A; Reddanna, Pallu; Radhakrishnan, Girish K

    2014-05-29

    Brucella spp. are facultative intracellular bacterial pathogens causing the zoonotic disease brucellosis. Here, we report the draft genome sequence of the Brucella melitensis strain from India designated Bm IND1, isolated from stomach contents of an aborted goat fetus.

  8. Draft Genome Sequence of the Field Isolate Brucella melitensis Strain Bm IND1 from India

    OpenAIRE

    Rao, Sashi Bhushan; Gupta, Vivek K.; Kumar, Mukesh; Hegde, Nagendra R; Splitter, Gary A.; Reddanna, Pallu; Radhakrishnan, Girish K.

    2014-01-01

    Brucella spp. are facultative intracellular bacterial pathogens causing the zoonotic disease brucellosis. Here, we report the draft genome sequence of the Brucella melitensis strain from India designated Bm IND1, isolated from stomach contents of an aborted goat fetus.

  9. 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....

  10. Evidence for co-infection of ovine prion strains in classical scrapie isolates.

    Science.gov (United States)

    Thackray, A M; Lockey, R; Beck, K E; Spiropoulos, J; Bujdoso, R

    2012-01-01

    The diversity of strains of ovine prions within classical scrapie isolates was investigated by transmission studies in wild type mice. To determine the maximum diversity of prion strains present in each ovine scrapie isolate examined, isolates from mice having the shortest and longest incubation times for terminal disease after primary inoculation were passaged serially. Serial passage of ARQ/ARQ scrapie isolates in RIII mice revealed the ME7 prion strain in mice with short incubation times for terminal prion disease and the 87A strain in those mice with long incubation times. Serial passage of VRQ/VRQ scrapie isolates in RIII mice led to emergence of the 221C prion strain in mice with short incubation times and a variant of the 221C strain in those mice with long incubation times. RIII mice with short incubation times had higher levels of total and proteinase K-resistant PrP(Sc) compared with those RIII mice with long incubation times, while mice with long incubation times had large aggregates and plaques of PrP(Sc). ME7 PrP(Sc) differed in stability compared with the 87A prion strain, while PrP(Sc) associated with 221C had similar stability to that of the 221C variant. Serial passage in VM mice led to identification of ME7 and 87V in the same scrapie isolate. The data show that different prion strains can emerge from the same ovine scrapie isolate following serial passage in wild type mice and that the transmission properties of these strains correlate with distinct patterns of PrP(Sc) deposition.

  11. Development of a systematic feedback isolation approach for targeted strains from mixed culture systems.

    Science.gov (United States)

    Poudel, Pramod; Tashiro, Yukihiro; Miyamoto, Hirokuni; Miyamoto, Hisashi; Okugawa, Yuki; Sakai, Kenji

    2017-01-01

    Elucidation of functions of bacteria in a mixed culture system (MCS) such as composting, activated sludge system is difficult, since the system is complicating with many unisolated bacteria. Here, we developed a systematic feedback isolation strategy for the isolation and rapid screening of multiple targeted strains from MCS. Six major strains (Corynebacterium sphenisci, Bacillus thermocloacae, Bacillus thermoamylovorans, Bacillus smithii, Bacillus humi, and Bacillus coagulans), which are detected by denaturing gradient gel electrophoresis (DGGE) analysis in our previous study on MCS for l-lactic acid production, were targeted for isolation. Based on information of suitable cultivation conditions (e.g., media, pH, temperature) from the literature, feedback isolation was performed to form 136 colonies. The following direct colony matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was optimised as the second screening to narrow down 20 candidate colonies from similar spectra patterns with six closest type strains. This step could distinguish bacteria at the species level with distance similarity scores ≥0.55 corresponding to 16S rRNA gene sequence similarity ≥98.2%, suggesting that this is an effective technique to minimize isolates close to targeted type strains. Analysis of 16S rRNA gene sequences indicated that two targeted strains and one strain related to the target had successfully been isolated, showing high similarities (99.5-100%) with the sequences from the DGGE bands, and that the other candidates were affiliated with three strains that were closely related to the target species. This study proposes a new method for systematic feedback isolation that may be useful for isolating targeted strains from MCS for further investigation.

  12. Genome Sequences of Two Copper-Resistant Escherichia coli Strains Isolated from Copper-Fed Pigs

    DEFF Research Database (Denmark)

    Lüthje, Freja L.; Hasman, Henrik; Aarestrup, Frank Møller;

    2014-01-01

    The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances.......The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances....

  13. Epitope characterization and variable region sequence of f1-40, a high-affinity monoclonal antibody to botulinum neurotoxin type a (Hall strain.

    Directory of Open Access Journals (Sweden)

    Miles C Scotcher

    Full Text Available BACKGROUND: Botulism, an often fatal neuroparalytic disease, is caused by botulinum neurotoxins (BoNT which consist of a family of seven serotypes (A-H produced by the anaerobic bacterium Clostridium botulinum. BoNT, considered the most potent biological toxin known, is a 150 kDa protein consisting of a 100 kDa heavy-chain (Hc and a 50 kDa light-chain (Lc. F1-40 is a mouse-derived, IgG1 monoclonal antibody that binds the light chain of BoNT serotype A (BoNT/A and is used in a sensitive immunoassay for toxin detection. We report the fine epitope mapping of F1-40 and the deduced amino acid sequence of the variable regions of the heavy and light chains of the antibody. METHODS AND FINDINGS: To characterize the binding epitope of F1-40, three complementary experimental approaches were selected. Firstly, recombinant peptide fragments of BoNT/A light-chain were used in Western blots to identify the epitope domains. Secondly, a peptide phage-display library was used to identify the specific amino acid sequences. Thirdly, the three-dimensional structure of BoNT/A was examined in silico, and the amino acid sequences determined from the phage-display studies were mapped onto the three-dimensional structure in order to visualize the epitope. F1-40 was found to bind a peptide fragment of BoNT/A, designated L1-3, which spans from T125 to L200. The motif QPDRS was identified by phage-display, and was mapped to a region within L1-3. When the three amino acids Q138, P139 and D140 were all mutated to glycine, binding of F1-40 to the recombinant BoNT/A light chain peptide was abolished. Q-138, P-139 and D-140 form a loop on the external surface of BoNT/A, exposed to solvent and accessible to F1-40 binding. CONCLUSIONS: The epitope of F1-40 was localized to a single exposed loop (ss4, ss5 on the Lc of BoNT. Furthermore amino acids Q138, P139 and D140 forming the tip of the loop appear critical for binding.

  14. Study of Antimicrobial Resistance of Acinetobacter Strains Isolated From Blood Cultures

    Directory of Open Access Journals (Sweden)

    H Zandi

    2007-06-01

    Full Text Available Background: Acinetobacter spp are associated with various nosocomial infections like as septicemia and are isolated form blood cultures in hospitalized patients. Methods: In this study, 45 Acinetobacter strains were isolated from blood samples in Yazd shahid sadoughi hospital from 21 March 2005 to 20 September 2006 and were identified by biochemical tests. Antibiotic susceptibility of the strains was tested by standard disk diffusion method. Results: In this research, 45 isolates identified as Acinetobacter and of isolated strains, 88.8% of them found sensitive to imipenem and 80% to ciprofloxacin. Also 51.5% to nalidixic Acid 24.5% to trimethoprim/sulphametoxazole, 11.1% to ceftazidim and ceftriaxone, 8.8% to cefotaxime and cefexime and also 6.6% to ceftizoxime. Conclusion: Because of increasing of drug resistance in Acinetobacter spp. Isolated from blood samples, it is necessary to perform susceptibility testing, also imipenem and ciprofloxacin recommended for drug therapy.

  15. Characterisation of Clostridium difficile strains isolated from Groote Schuur Hospital, Cape Town, South Africa.

    Science.gov (United States)

    Kullin, B; Brock, T; Rajabally, N; Anwar, F; Vedantam, G; Reid, S; Abratt, V

    2016-10-01

    The C. difficile infection rate in South Africa is concerning. Many strains previously isolated from diarrhetic patients at Groote Schuur Hospital were ribotype 017. This study further characterised these strains with respect to their clonal relationships, antibiotic susceptibility, toxin production and various attributes impacting on pathogen colonisation. Multilocus variable-number tandem-repeat analysis (MLVA) was used to characterise all C. difficile isolates. Antibiotic susceptibility was determined by E-test and PCR-based analysis of the ermB, gyrA and gyrB genes. Auto-aggregation of cells was measured in broth, and biofilm formation observed in 24-well plates. Toxins were measured using the Wampole C DIFF TOX A/B II kit. Most isolates belonged to the ribotype 017 group. Identical MLVA types occurred in different wards over time, and several patients were infected with identical strains. All isolates were susceptible to vancomycin and metronidazole, but some ribotype 017 isolates showed reduced metronidazole susceptibility (≥2 mg l(-1)). Sixty-nine percent of ribotype 017 isolates were resistant to moxifloxacin, and 94 % to erythromycin, compared to 0 % and 17 % resistance, respectively, in non-ribotype 017 isolates. The ermB gene and mutations in the gyrA and/or gyrB genes were linked to erythromycin and moxifloxacin resistance, respectively. Ribotype 017 isolates auto-aggregated more strongly than other isolates and produced lower levels of the TcdB toxin than a reference strain. Certain strains produced strong biofilms. Patient-to-patient transfer and unique infection events could cause the predominance of ribotype 017 strains in the cohort. Multi-drug resistant strains are a potential reservoir for future infections.

  16. 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.

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

    Science.gov (United States)

    Sørensen, Martine C Holst; Gencay, Yilmaz Emre; Birk, Tina; Baldvinsson, Signe Berg; Jäckel, Claudia; Hammerl, Jens A; Vegge, Christina S; Neve, Horst; Brøndsted, Lone

    2015-01-01

    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.

  18. Molecular subtyping of Clostridium botulinum by pulsed-field gel electrophoresis.

    Science.gov (United States)

    Lúquez, Carolina; Joseph, Lavin A; Maslanka, Susan E

    2015-01-01

    Pulsed-field gel electrophoresis (PFGE) has been extensively used to estimate the genetic diversity of Clostridium botulinum. In addition, PFGE is the standard method for investigating foodborne outbreaks associated with various enteric pathogens, including C. botulinum. PFGE can be used to exclude a suspected but not confirmed food source when the patterns of the food and clinical isolates are different. Indistinguishable PFGE patterns may also be useful for linking isolates between patients or to a food source, but results must be interpreted within an epidemiological context to ensure isolates are truly related. Here, we describe a standardized laboratory protocol for molecular subtyping of C. botulinum by PFGE.

  19. Multilocus sequence typing reveals that Bacillus cereus strains isolated from clinical infections have distinct phylogenetic origins.

    Science.gov (United States)

    Barker, Margaret; Thakker, Bishan; Priest, Fergus G

    2005-04-01

    Eight strains of Bacillus cereus isolated from bacteremia and soft tissue infections were assigned to seven sequence types (STs) by multilocus sequence typing (MLST). Two strains from different locations had identical STs. The concatenated sequences of the seven STs were aligned with 65 concatenated sequences from reference STs and a neighbor-joining tree was constructed. Two strains were distantly related to all reference STs. Three strains were recovered in a clade that included Bacillus anthracis, B. cereus and rare Bacillus thuringiensis strains while the other three strains were assigned to two STs that were more closely affiliated to most of the B. thuringiensis STs. We conclude that invasive B. cereus strains do not form a single clone or clonal complex of highly virulent strains.

  20. 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.

  1. Cross-comparison of leaching strains isolated from two different regions: Chambishi and Dexing copper mines.

    Science.gov (United States)

    Ngom, Baba; Liang, Yili; Liu, Xueduan

    2014-01-01

    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.

  2. Genetic and Filogenetic Characterization of some Newcastle Strains Isolated from Poultry in Albania

    Directory of Open Access Journals (Sweden)

    MARSEL BORAKAJ

    2015-06-01

    Full Text Available Abstract section. In this study, we present the molecular characterization and phylogenetic analysis of three strains of NDV, isolated from the Tirana region in Albania during the 2011-2014 years. Three strains with number 28, 29 and 31, isolated from a different farm of poultry in Tirana Region (Rural flocks, which were diagnosed clinically with the ND. The Intracerebral Pathogenicity Index in SPF bird one day old was determined by doing the proteolytic sequencing at the cleavage, and specifying the aminoacid motif at proteolytic cleavage site. More over we performed BLAST search and phylogenetic analysis of obtained RNA sequences. All strains replicated well in the SPF –chicken emryo eggs. The isolates displayed an aminoacid motif at the proteolytic cleavage site at the Fusion (F protein with multiple basic amino acids as a well a Phenylalanine on position 117. For one isolate (28 numerous nucleotide positions had signals for at last two nucleotides, making it imposible to conclude on a specific sequence. The pathogenicity of all three isolates (28, 29 and 33, was assessed by the analysis of the F protein cleavage site and by standart ICPI. The ICPI (pathogenicity index of our strains varies from of 1.85, 2 and 1.75, respectively which according [19,7] are typical for velogenic strains of NDV. We found that two NDV strain has a most close genetic relationship with the Serbia 2007 NDV, having 98% similarity at nucleotide level.Velogenic viscerotropic strains are considered endemic in our country.

  3. 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.

  4. [Antibiotic resistance of Shigella strains isolated in Cádiz (author's transl)].

    Science.gov (United States)

    García Martos, P; Marín Casanova, P; García Herruzo, J; Fernández Gutiérrez del Alamo, C

    1980-12-15

    The incidence of shigellosis at the Residencia Sanitaria Fernando Zamacola (Cáciz, Spain) and the antibiotic sensitivity of 94 strains of Shigella sonnei and 40 strains of Shigella flexneri, isolated during the year 1979, has been studied taking into account the present status of strain resistance to the major antibiotics. Three epidemic bouts of shigellosis were detected: one in february by Shigella sonnei (16 cases), and two others in august-september and november due to Shigella flexneri (43 and 29 cases). Children 2 to 5 years old had the highest incidence of Shigella infection. Almost all strains isolated were resistant to the sulphonamides (99.77%). Ampicillin and chloramphenicol had little efficacy against Shigella flexneri (95.00 and 92.50% resistance). The percent resistance of Shigella sonnei strains to phosphomycin was elevated (44.69%). All strains studied were sensitive to colimycin and showed little resistance to the combination trimethoprim-sulfamethoxazol (16.42%).

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

    Institute of Scientific and Technical Information of China (English)

    HUANG BaoLing; L(U) ChengQun; WU Bo; FAN LiQin

    2007-01-01

    Symbiotic nitrogen fixation of rhizobia and leguminous plants is considered as the most important biologic nitrogen fixation system on earth. Symbiotic nodulafion 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.

  6. A rhizobia strain isolated from root nodule of gymno-sperm Podocarpus macrophyllus

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    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 macro-phyllus 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.

  7. 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

  8. 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

  9. Draft Genome Sequence of Pedobacter sp. Strain Hv1, an Isolate from Medicinal Leech Mucosal Castings.

    Science.gov (United States)

    Ott, Brittany M; Beka, Lidia; Graf, Joerg; Rio, Rita V M

    2015-12-17

    The Pedobacter sp. Hv1 strain was isolated from the medicinal leech, Hirudo verbana, mucosal castings. These mucosal sheds have been demonstrated to play a role in horizontal symbiont transmission. Here, we report the draft 4.9 Mbp genome sequence of Pedobacter sp. strain Hv1.

  10. Genome Sequences of 14 Firmicutes Strains Isolated from the Human Vagina.

    Science.gov (United States)

    Deitzler, Grace E; Ruiz, Maria J; Weimer, Cory; Park, SoEun; Robinson, Lloyd; Hallsworth-Pepin, Kymberlie; Wollam, Aye; Mitreva, Makedonka; Lewis, Amanda L; Lewis, Warren G

    2016-01-01

    Research on vaginal infections is currently limited by a lack of available fully sequenced bacterial reference strains. Here, we present strains (now available through BEI Resources) and genome sequences for a set of 14 vaginal isolates from the phylum Firmicutes These genome sequences provide a valuable resource for future research in understanding the role of Gram-positive bacteria in vaginal health and disease.

  11. Plasmid profiling and antibiotics resisitance of Escherichia coli strains isolated from Mytilus galloprovincialis and seawater

    Directory of Open Access Journals (Sweden)

    Cumhur Avşar

    2014-09-01

    Full Text Available Objective: To investigate plasmid DNA profiles and the antibiotic resistance of a total of 41 strains of Escherichia coli (E. coli isolated from seawater and mussel collected from 15 different sampling stations in Sinop, Turkey. Methods: Most probable number technique was used for detection of E. coli. Antibiotic susceptibilities of the isolates were determined by the disc diffusion method. Plasmid DNA of the strains was extracted by the alkaline lyses procedure. Results: According to morphological and physiological properties, it was determined that the isolates belonged to E. coli species. Antibiotic susceptibility of the strains was determined against seven standard drugs using disc diffusion method. All isolates were resistant to bacitracin (100%, novobiocin (100%, ampicillin (12.5%, tetracycline (7.5%, ceftazidime (5% and imipenem (2.5%, respectively, whereas the strains were susceptible to polymyxin B (100%. The multiple antibiotic resistance values for the strains were found in range from 0.28 to 0.57. In addition, plasmid DNA analyses results confirmed that 22 strains harbored a single or more than two plasmids sized approximately between 24.500 to 1.618 bp. The high-size plasmid (14.700 bp was observed as common in 21 of all strains. Conclusions: As a result, our study indicated that the presence of antibiotic resistant E. coli strains in seawater and mussel might be potential risk for public health issue.

  12. Draft Genome Sequence of Clostridium tyrobutyricum Strain DIVETGP, Isolated from Cow's Milk for Grana Padano Production

    OpenAIRE

    2015-01-01

    We announce the draft genome sequence of Clostridium tyrobutyricum strain DIVETGP. This strain was isolated from cow’s milk used for Grana Padano cheese production. The genome was obtained using Illumina HiSeq technology and comprises 45 contigs for 3,018,999 bp, with a G+C content of 30.8%.

  13. Draft Genome Sequence of Clostridium tyrobutyricum Strain DIVETGP, Isolated from Cow's Milk for Grana Padano Production.

    Science.gov (United States)

    Soggiu, Alessio; Piras, Cristian; Gaiarsa, Stefano; Bendixen, Emøke; Panitz, Frank; Bendixen, Christian; Sassera, Davide; Brasca, Milena; Bonizzi, Luigi; Roncada, Paola

    2015-03-26

    We announce the draft genome sequence of Clostridium tyrobutyricum strain DIVETGP. This strain was isolated from cow's milk used for Grana Padano cheese production. The genome was obtained using Illumina HiSeq technology and comprises 45 contigs for 3,018,999 bp, with a G+C content of 30.8%.

  14. Draft Genome Sequence of Clostridium tyrobutyricum Strain DIVETGP, Isolated from Cow's Milk for Grana Padano Production

    DEFF Research Database (Denmark)

    Soggiu, Alessio; Piras, Cristian; Gaiarsa, Stefano;

    2015-01-01

    We announce the draft genome sequence of Clostridium tyrobutyricum strain DIVETGP. This strain was isolated from cow's milk used for Grana Padano cheese production. The genome was obtained using Illumina HiSeq technology and comprises 45 contigs for 3,018,999 bp, with a G+C content of 30.8%....

  15. Genome Sequence of Marinobacter sp. Strain MCTG268 Isolated from the Cosmopolitan Marine Diatom Skeletonema costatum.

    Science.gov (United States)

    Gutierrez, Tony; Whitman, William B; Huntemann, Marcel; Copeland, Alex; Chen, Amy; Kyrpides, Nikos; Markowitz, Victor; Pillay, Manoj; Ivanova, Natalia; Mikhailova, Natalia; Ovchinnikova, Galina; Andersen, Evan; Pati, Amrita; Stamatis, Dimitrios; Reddy, T B K; Ngan, Chew Yee; Chovatia, Mansi; Daum, Chris; Shapiro, Nicole; Cantor, Michael N; Woyke, Tanja

    2016-09-08

    Marinobacter sp. strain MCTG268 was isolated from the cosmopolitan marine diatom Skeletonema costatum and can degrade oil hydrocarbons as sole sources of carbon and energy. Here, we present the genome sequence of this strain, which is 4,449,396 bp with 4,157 genes and an average G+C content of 57.0%.

  16. [Occurrence and antimicrobial susceptibility of Morganella morganii strains isolated from clinical samples].

    Science.gov (United States)

    Zalas-Wiecek, Patrycja; Gospodarek, Eugenia; Wróblewska, Joanna

    2012-01-01

    The aim of this study was the evaluation of occurrence and antimicrobial susceptibility of M morganii rods isolated from clinical samples. This study included 201 strains isolated in the Clinical Microbiology Department of Dr. A. Jurasz University Hospital in 2008-2010. Identification to species was carried out on the basis of the results of biochemical reactions included in the tests ID 32E and VITEK2 GN. Antimicrobial susceptibility of M. morganii rods was determined by the disk-diffusion method on Mueller-Hinton II Agar. Strains of M morganii most commonly isolated from skin and soft tissue, and material taken from the urinary tract, mainly from patients of Anesthesiology and Intensive Care Unit, Department of General and Vascular Surgery and Department of General Surgery and Endocrinology. All of M morganii strains isolated during the three years were susceptible to carbapenems. We reported decrease of strains susceptible to piperacillin and chloramphenicol. In 2010 we showed a higher percentage of strains intermediate to tigecycline, compared with 2009. We observed increase in the percentage of strains resistant to cefoperazone with sulbactam and reported decrease in the percentage of strains resistant and intermediate to aminoglycosides. Extended Spectrum Beta-Lactamases were produced by 13 (6,5%) of M morganii strains.

  17. Plasmid profiling and antibiotics resisitance of Escherichia coli strains isolated from Mytilus galloprovincialis and seawater

    Institute of Scientific and Technical Information of China (English)

    Cumhur Avşar; İsmet Berber

    2014-01-01

    Objective: To investigate plasmid DNA profiles and the antibiotic resistance of a total of 41 strains of Escherichia coli (E. coli) isolated from seawater and mussel collected from 15 different sampling stations in Sinop, Turkey. Methods: Most probable number technique was used for detection of E. coli. Antibiotic susceptibilities of the isolates were determined by the disc diffusion method. Plasmid DNA of the strains was extracted by the alkaline lyses procedure.Results:According to morphological and physiological properties, it was determined that the isolates belonged to E. coli species. Antibiotic susceptibility of the strains was determined against seven standard drugs using disc diffusion method. All isolates were resistant to bacitracin (100%), novobiocin (100%), ampicillin (12.5%), tetracycline (7.5%), ceftazidime (5%) and imipenem (2.5%), respectively, whereas the strains were susceptible to polymyxin B (100%). The multiple antibiotic resistance values for the strains were found in range from 0.28 to 0.57. In addition, plasmid DNA analyses results confirmed that 22 strains harbored a single or more than two plasmids sized approximately between 24.500 to 1.618 bp. The high-size plasmid (14.700 bp) was observed as common in 21 of all strains.Conclusions:As a result, our study indicated that the presence of antibiotic resistant E. coli strains in seawater and mussel might be potential risk for public health issue.

  18. Genetic analysis of Streptococcus agalactiae strains isolated from neonates and their mothers.

    NARCIS (Netherlands)

    Melchers, W.J.G.; Bakkers, J.M.J.E.; Toonen, M.; Kuppeveld, F.J.M. van; Trijbels-Smeulders, M.J.A.M.; Hoogkamp-Korstanje, J.A.A.

    2003-01-01

    Streptococcus agalactiae or group B streptococcus (GBS) is the most common cause of neonatal sepsis and meningitis in neonates. One of the major questions is whether the GBS strains able to cause neonatal invasive disease have peculiar genetic features. A collection of S. agalactiae strains, isolate

  19. 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

  20. Draft Genome Sequence of Deinococcus sp. Strain RL Isolated from Sediments of a Hot Water Spring.

    Science.gov (United States)

    Mahato, Nitish Kumar; Tripathi, Charu; Verma, Helianthous; Singh, Neha; Lal, Rup

    2014-07-17

    Deinococcus sp. strain RL, a moderately thermophilic bacterium, was isolated from sediments of a hot water spring in Manikaran, India. Here, we report the draft genome (2.79 Mbp) of this strain, which contains 62 contigs and 2,614 coding DNA sequences, with an average G+C content of 69.4%.

  1. 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.

  2. Draft Genome Sequences of Four Enterococcus faecium Strains Isolated from Argentine Cheese

    Science.gov (United States)

    Martino, Gabriela P.; Quintana, Ingrid M.; Espariz, Martín; Blancato, Victor S.; Gallina Nizo, Gabriel; Esteban, Luis

    2016-01-01

    We report the draft genome sequences of four Enterococcus faecium strains isolated from Argentine regional cheeses. These strains were selected based on their technological properties, i.e., their ability to produce aroma compounds (diacetyl, acetoin, and 2,3-butanediol) from citrate. The goal of our study is to provide further genetic evidence for the rational selection of enterococci strains based on their pheno- and genotype in order to be used in cheese production. PMID:26847907

  3. 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.

  4. An innovative molecular detection tool for tracking and tracing Clostridium botulinum types A, B, E, F and other botulinum neurotoxin producing Clostridia based on the GeneDisc cycler.

    Science.gov (United States)

    Fach, P; Fenicia, L; Knutsson, R; Wielinga, P R; Anniballi, F; Delibato, E; Auricchio, B; Woudstra, C; Agren, J; Segerman, B; de Medici, D; van Rotterdam, B J

    2011-03-01

    Rapid and specific detection of botulinum neurotoxin (BoNT) producing Clostridia is a priority for public health authorities, in case of both natural and intentional botulism outbreaks. This study reports on the evaluation of a detection system based on the GeneDisc Cycler designed for simultaneously testing the bont/A, bont/B, bont/E and bont/F genes encoding for the botulinum neurotoxins types A, B, E and F. BoNT-producing Clostridia (n = 102) and non-BoNT-producing bacteria (n = 52) isolated from clinical, food and environmental samples were tested using this macro-array and results were compared to the reference lethality test on mice. The bont genes were correctly detected in all C. botulinum type A, B, E and F strains available, as well as in toxigenic C. baratii type F and toxigenic C. butyricum type E. No cross reactivity was observed with non human-toxigenic bacteria, C. botulinum types C, D and G. The identification of the bont genotype using the macro-array was correlated to toxino-typing of the BoNTs as determined by the mouse bioassay. An "evaluation trial" of the GeneDisc array performed blind in four European laboratories with 77 BoNT-producing Clostridia as well as 10 food and clinical samples showed that the developed macro-array is specific and reliable for identifying BoNT/A-, BoNT/B-, BoNT/E- and BoNT/F-producing clostridial strains and for screening naturally contaminated food and fecal samples. The test is robust, has a low detection limit (c.a. 5 to 50 genome copies in the PCR reaction microwell) and is promising for monitoring BoNT-producing Clostridia in different kinds of samples including food and clinical samples.

  5. Hemagglutinin gene shuffling among Clostridium botulinum serotypes C and D yields distinct sugar recognition of the botulinum toxin complex.

    Science.gov (United States)

    Miyata, Keita; Suzuki, Tomonori; Hayashi, Shintaro; Miyashita, Shin-Ichiro; Ohyama, Tohru; Niwa, Koichi; Watanabe, Toshihiro; Sagane, Yoshimasa

    2015-10-01

    Clostridium botulinum strains produce a large-sized toxin complex (TC) that is composed of botulinum neurotoxin (BoNT), non-toxic non-hemagglutinin and three different hemagglutinins (HA-70, HA-33 and HA-17). HA components enhance toxin delivery across the intestinal cell wall in a sugar chain-dependent manner. Here we characterized the sugar recognition of serotype D strain 1873 (D-1873) botulinum L-TC. Most L-TCs produced by serotype C and D strains bind to cells via interactions between HA-33 and cell surface sialo-oligosaccharides. However, like the previously reported L-TC produced by serotype C strain Yoichi (C-Yoichi), D-1873 L-TC binds only to cells that have been treated with neuraminidase, indicating that they recognize asialo-oligosaccharides. The D-1873 HA-33 amino acid sequence is similar to that of C-Yoichi, but had lower similarity to the majority of serotype C and D HA-33s. A comparison of TC component primary structures for 12 serotype C and D strains suggested that at least three types of HA-33 genes exist, and these are shuffled among the serotype C and D strains independently of BoNT serotype. This shuffling produces the distinct sugar recognition of serotype C and D botulinum TCs.

  6. Protease activity in Giardia duodenalis trophozoites of axenic strains isolated from symptomatic and asymptomatic patients

    Directory of Open Access Journals (Sweden)

    Guimarães Semíramis

    2003-01-01

    Full Text Available We have examined by gelatin-SDS-PAGE the protease activity in cell lysates of Giardia duodenalis trophozoites of two axenic strains isolated in Brazil from a symptomatic patient (BTU-11 and an asymptomatic carrier (BTU-10, and the reference strain Portland 1 (P1. The proteolysis band patterns showed differences among strains isolated from asymptomatic and symptomatic individuals. The lysate of the strain BTU-10, showed only five hydrolysis bands, while a greater number of bands (10-11 bands was seen in strains BTU-11 and P1. The protease activity in all lysates was inhibited by cysteine (E-64 and iodoacetamide and serine proteases (TPCK and TLCK inhibitors, but not by PMSF and EDTA. In general, the results revealed protease activities in G. duodenalis trophozoites of Brazilian axenic strains and the predominance of cysteine proteinases. It should be stressed the inter-strain difference in hydrolysis band patterns observed between strains isolated from symptomatic patients and the strain obtained from an asymptomatic carrier.

  7. 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....

  8. 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.

  9. Detection of diarrheagenic Escherichia coli strains isolated from dogs and cats in Brazil.

    Science.gov (United States)

    Puño-Sarmiento, Juan; Medeiros, Leonardo; Chiconi, Carolina; Martins, Fernando; Pelayo, Jacinta; Rocha, Sérgio; Blanco, Jorge; Blanco, Miguel; Zanutto, Marcelo; Kobayashi, Renata; Nakazato, Gerson

    2013-10-25

    Escherichia coli are gut microbiota bacteria that can cause disease in some humans and other animals, including dogs and cats that humans often keep as pets. Diarrheagenic E. coli (DEC) strains are classified into six categories: enteropathogenic (EPEC), enterotoxigenic (ETEC), Shiga toxin-producing (STEC), enteroinvasive (EIEC), enteroaggregative (EAEC), and diffuse-adhering E. coli (DAEC). In this study 144 and 163 E. coli colonies were isolated from the fecal samples of 50 dogs and 50 cats, respectively, with and without diarrhea from a Veterinary Hospital (clinical isolates). The virulence factors were determined using multiplex Polymerase Chain Reaction. Adherence assays, antibacterial susceptibility and serotyping (somatic or flagellar antigens) were performed on DEC isolates. We found 25 (17.4%) and 4 (2.5%) DEC strains isolated from dogs and cats, respectively. Only the EPEC and EAEC pathotypes were found in both animals. Meanwhile, genes from other pathotypes (STEC, EIEC, and ETEC) were not found in these clinical isolates. All of the DEC strains showed mannose-resistant adherence to HEp-2 and HeLa cells, and aggregative adherence was predominant in these isolates. Multiresistant strains to antimicrobials were found in most DEC strains including usual and unusual antimicrobials in veterinary practices. The serotypes of these DEC isolates were variable. The ONT serotype was predominant in these isolates. Some serotypes found in our study were described to human DEC. Here, we demonstrate that pets carry virulent DEC genes, which are mainly strains of EPECs and EAECs. The presence of these virulence factors in isolates from animals without diarrhea suggests that pets can act as a reservoir for human infection.

  10. EMERGENCE OF MULTI DRUG RESISTANT STRAINS OF E. COLI ISOLATED FROM URINARY TRACT INFECTION IN NAMAKKAL

    OpenAIRE

    2012-01-01

    In this study we evaluated the multi drug resistant strains of Escherichia coli isolated from urinary tract infection in different age groups in Namakkal. Totally 153 isolates of E.coli were obtained from 700 urine samples. Present study indicates that there is a high prevalence of multidrug resistant E. coli in female 24.34% followed by male 18.14%. After confirmation, isolates were performed to resistance patterns of different antibiotics were determined by standard disk diffusion method. T...

  11. 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.

  12. Mating Type Gene (MAT) and Itraconazole Susceptibility of Trichophyton tonsurans Strains Isolated in Japan.

    Science.gov (United States)

    Hiruma, Junichiro; Okubo, Miki; Kano, Rui; Kumagawa, Mai; Hiruma, Masataro; Hasegawa, Atsuhiko; Kamata, Hiroshi; Tsuboi, Ryoji

    2016-06-01

    Infection by Trichophyton tonsurans is an emerging fungal epidemic in Japan. Itraconazole (ITZ) and terbinafine have been used for the treatment of this infection for 15 years. However, patients with T. tonsurans infections have been shown to remain uncured or to become reinfected, suggesting that subclinical infection or polyphyletic strains and/or antifungal drug-resistant strains might be occurring in Japan. In this study, PCR analysis was performed to confirm the presence of the mating type locus MAT in genomic DNA from 60 Japanese clinical isolates of T. tonsurans, and to assess the previously postulated clonal origin of clinical isolates of this species. Antifungal susceptibility testing on isolates also was performed to confirm the absence of strains resistant to ITZ. PCR analysis proved that all 60 strains contained the MAT1-1 allele, while none contained the MAT1-2 allele. As determined by E-test, the mean MIC of ITZ in the 60 strains was 0.023 mg/L (range 0.002-0.125 mg/L). All strains of T. tonsurans isolated in Japan were clonal and were not resistant to ITZ. Therefore, dermatophytosis due to T. tonsurans is expected to respond to ITZ, since clinical isolates of T. tonsurans tested to date have been susceptible to this antifungal. This infection is proliferating as a subclinical infection in Japan.

  13. 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).

  14. [Isolation and characterization of wild Sporothrix schenkii strains and investigation of sporototrichin reactors].

    Science.gov (United States)

    Sánchez-Alemán, Miguel Angel; Araiza, Javier; Bonifaz, Alexandro

    2004-01-01

    We conducted a study in the southern mountains of the Mexican State of Oaxaca that consisted of isolation of wild Sporothrix schenckii strains obtained from soil samples and investigation of positive reactors to skin test reaction with sprotrichin antigen. The study was conducted by means of recollection of soil samples and processing of these with dilution methods and fungal isolation in ordinary culture media Sabouraud simple Agar with and without antibiotics (SS, SA). Suspected strains underwent dimorphism, melanin formation, and virulence confirmation tests. Investigation of positive reactors to sporotrichin Y (yeast) was also conducted. Three supposed strains were identified due to their reproductive characteristics, melanin production, and virulence. In the community, 144 individuals were studied, of whom 6.25% were positive to sporotichin. Isolation of virulent strains of Sporothrix schenkii from nature (soil) and primoinfection of a percentage of the studied population were confirmed.

  15. Primary Isolation Strain Determines Both Phage Type and Receptors Recognised by Campylobacter jejuni Bacteriophages

    DEFF Research Database (Denmark)

    Sørensen, Martine C. Holst; Gencay, Yilmaz Emre; Birk, Tina

    2015-01-01

    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...

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

    Directory of Open Access Journals (Sweden)

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

    Science.gov (United States)

    Rahman, Mustafizur; Jubair, Mohammad; Alam, Meer T; Weppelmann, Thomas A; Azarian, Taj; Salemi, Marco; Sakharuk, Ilya A; Rashid, Mohammed H; Johnson, Judith A; Yasmin, Mahmuda; Morris, J Glenn; Ali, Afsar

    2014-01-01

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Unexpected detection of animal VP7 genes among common rotavirus strains isolated from children in Mexico.

    Science.gov (United States)

    Laird, A R; Ibarra, V; Ruiz-Palacios, G; Guerrero, M L; Glass, R I; Gentsch, J R

    2003-09-01

    In the course of characterizing 103 rotaviruses from children in Mexico, we found that the majority of strains were globally common types (55.4% of total), while uncommon types represented 5.7%, mixed infections with common types represented 14.8%, and partially or fully nontypeable isolates represented about 24%. Serotype G9 was detected for the first time in Mexico. We sequenced a subset of strains that were G nontypeable by reverse transcriptase PCR and found surprisingly that two strains having common human rotavirus P genotypes (8 and 6) had serotype G3 and G4 VP7 gene sequences that shared closer homology with canine and porcine strains, respectively, than with human strains, suggesting that these isolates represented reassortants between human and animal rotaviruses.

  2. Phenotypic and genotypic characterization of Xanthomonas campestris strains isolated from cabbage, kale and broccoli

    Directory of Open Access Journals (Sweden)

    Popović Tatjana

    2013-01-01

    Full Text Available Thirty-six strains of Xanthomonas campestris pv. campestris (Xcc isolated from cabbage, kale and broccoli were identified according to their pathogenicity, phenotypic and genotypic characterization. Pathogenicity was confirmed by the injection method with a hypodermic syringe into the mesophilic tissue of cabbage leaves. All strains were Gramnegative, aerobic, catalase-positive, oxidase-negative, grew at 35°C, produced levan, H2S and indole, did not reduce nitrate, hydrolyzed Tween 80, starch, gelatin and esculin and did not show tolerance to 0.1 and 0.02% TTC. The strains produced acid from d-arabinose, arginine, dulcitol, galactose, d-glucose, maltose, mannose, sorbitol, sucrose and xylose. The genetic characterization was based on the sequence analyses of 16S rDNA and ERIC and BOX PCR. Strains of different pathovars were also used to compare PCR resulting patterns. BOX-PCR of the strains from kale and broccoli, obtained using (GTG5 primer, yielded patterns with a high similarity level to pathovar reference strain Xcc. The strains from cabbage yielded BOX and ERIC product patterns, distinguishing them from the other tested strains and reference strains. 16S rDNA of the representative strains was closely related to Xcc strain ATCC 33913. ERIC PCR and BOX using (GTG5 primer generated different Xcc patterns and were effective in distinguishing strains from different plant hosts. [Projekat Ministarstva nauke Republike Srbije, br. III43010 i br. III46007

  3. Systemic neonatal candidosis: the karyotyping of Candida albicans strains isolated from neonates and health-workers.

    Science.gov (United States)

    Ben Abdeljelil, J; Ben Saida, N; Saghrouni, F; Fathallah, A; Boukadida, J; Sboui, H; Ben Said, M

    2010-01-01

    Candida albicans has become an important cause of nosocomial infections in neonatal intensive care units (NICUs). The aim of the present study was to compare C. albicans strains isolated from neonates (NN) suffering from systemic candidosis and from nurses in order to determine the relatedness between NN and health workers' strains. Thirty-one C. albicans strains were isolated from 18 NN admitted to the NICU of the neonatology service of Farhat Hached Hospital of Sousse, Tunisia and suffering from systemic candidosis, together with five strains recovered from nurses suffering from C. albicans onychomycosis. Two additional strains were tested, one from an adult patient who developed a systemic candidosis and the second from an adult with inguinal intertrigo. All strains were karyotyped by pulsed-field gel electrophoresis (PFGE) with a CHEF-DR II system. Analysis of PFGE patterns yielded by the 38 strains tested led to the identification of three pulsotypes that were designated I, II and III, and consisted of six chromosomal bands with a size ranging from 700 to >2500 kbp. The most widespread was the pulsotype I, which was shared by 17 NN and the five nurses' strains. The identity between NN and nurses' strains is very suggestive of a nosocomial acquisition from health-workers.

  4. EMERGENCE OF MULTI DRUG RESISTANT STRAINS OF E. COLI ISOLATED FROM URINARY TRACT INFECTION IN NAMAKKAL

    Directory of Open Access Journals (Sweden)

    P. Poongothai

    2012-08-01

    Full Text Available In this study we evaluated the multi drug resistant strains of Escherichia coli isolated from urinary tract infection in different age groups in Namakkal. Totally 153 isolates of E.coli were obtained from 700 urine samples. Present study indicates that there is a high prevalence of multidrug resistant E. coli in female 24.34% followed by male 18.14%. After confirmation, isolates were performed to resistance patterns of different antibiotics were determined by standard disk diffusion method. The antibiogram patterns of the isolates showed a high percentage of multidrug resistance to cephodoxime, Novobiocin and vancomycin which was repeated to most of the isolates. Fifty resistance patterns were recognized among the MDR strains. This study revealed that E. coli was the predominant bacterial pathogen of community acquired UTIs in Aligarh, India. It also demonstrated an increasing resistance to number of antibiotics. This study is useful for clinician in order to improve the empirical treatment.

  5. 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.

  6. Molecular characterization of an apoptotic strain of Newcastle disease virus isolated from an outbreak in India.

    Science.gov (United States)

    Kumar, U; Kumar, S

    2015-08-01

    Newcastle disease (ND) is a highly contagious disease of poultry. The ND virus (NDV) encodes an error-prone RNA-dependent RNA polymerase which can cause high mutation rate leading to the emergence of its new antigenic variants. Antigenic difference of NDV strains may result in massive outbreak in vaccinated and unvaccinated poultry flocks around the globe. Apart from its pathogenic potential NDV has been explored as an oncolytic agent for a broad range of human cancers. In the present study, we isolated a novel NDV strain from an outbreak in chicken flock from the eastern part of India. Molecular characterization showed the NDV strain to be virulent in nature. The complete genome sequence analysis of the newly isolated strains belongs to genotype XIII. Moreover, the newly isolated strain of NDV showed positive results in various apoptotic assays in human breast cancer cells, MCF-7. The study will be useful to explore the possibility of using a newly isolated strain of NDV for virotherapy.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. Biodegradation of Ochratoxin A by Bacterial Strains Isolated from Vineyard Soils

    Directory of Open Access Journals (Sweden)

    Palmira De Bellis

    2015-11-01

    Full Text Available Ochratoxin A (OTA is a mycotoxin with a main nephrotoxic activity contaminating several foodstuffs. In the present report, five soil samples collected from OTA-contaminated vineyards were screened to isolate microorganisms able to biodegrade OTA. When cultivated in OTA-supplemented medium, OTA was converted in OTα by 225 bacterial isolates. To reveal clonal relationships between isolates, molecular typing by using an automated rep-PCR system was carried out, thus showing the presence of 27 different strains (rep-PCR profiles. The 16S-rRNA gene sequence analysis of an isolate representative of each rep-PCR profiles indicated that they belonged to five bacterial genera, namely Pseudomonas, Leclercia, Pantoea, Enterobacter, and Acinetobacter. However, further evaluation of OTA-degrading activity by the 27 strains revealed that only Acinetobacter calcoaceticus strain 396.1 and Acinetobacter sp. strain neg1, consistently conserved the above property; their further characterization showed that they were able to convert 82% and 91% OTA into OTα in six days at 24 °C, respectively. The presence of OTα, as the unique OTA-degradation product was confirmed by LC-HRMS. This is the first report on OTA biodegradation by bacterial strains isolated from agricultural soils and carried out under aerobic conditions and moderate temperatures. These microorganisms might be used to detoxify OTA-contaminated feed and could be a new source of gene(s for the development of a novel enzymatic detoxification system.

  12. Isolation and screening of strains producing high amounts of rutin degrading enzymes from Fagopyrum tataricum seeds.

    Science.gov (United States)

    Zheng, Ya-Di; Luo, Qing-Lin; Zhou, Mei-Liang; Wang, De-Zhou; Zhang, Ye-Dong; Shao, Ji-Rong; Zhu, Xue-Mei; Tang, Yu

    2013-02-01

    The rutin degrading enzyme (RDE) was isolated and purified from tartary buckwheat seeds. The RDE was purified about 11.34-fold and its final yield was 3.5%, which was very low, due to our purification strategy of giving priority to purity over yield. The RDE molecular weight was estimated to be about 60 kDa. When rutin was used as substrate, an optimal enzyme activity was seen at around pH 5.0 and 40 °C. Strains isolation strategy characterized by the use of rutin as sole carbon source in enrichment cultures was used to isolate RDE-producing strains. Then the active strains were identified by morphology characterization and 18s rDNA-ITS (Internal Transcribed Spacer) gene sequencing. Three isolates coded as B3, W2, Y2 were successfully isolated from fusty Fagopyrum tataricum flour cultures. Strain B3 possessed the highest unit activity among these three strains, and its total activity reached up to 171.0 Unit. The active isolate (B3) could be assigned to Penicillium farinosum. When the Penicillium farinosum strains were added to tartary buckwheat flour cultures at pH 5.0, 30 °C after 5 days fermentation, the quercetin production raised up to 1.78 mg/l, almost 5.1 times higher than the fermentation without the above active strains. Hence, a new approach was available to utilize microorganism-aided fermentation for effective quercetin extraction from Fagopyrum tataricum seeds.

  13. Antimicrobial activities of rhizobacterial strains of Pseudomonas and Bacillus strains isolated from rhizosphere soil of carnation (Dianthus caryophyllus cv. Sunrise).

    Science.gov (United States)

    Sharma, Sapna; Kaur, Mohinder

    2010-06-01

    Under the present study, an attempt was made to characterize rhizobacteria i.e. Pseudomonas and Bacillus species isolated from rhizosphere of carnation to evaluate their growth promoting effect on carnation so as to select and develop more efficient indigenous plant growth promoting and disease suppressing bioagents of specific soil type and specific plant type. Maximum strains of Pseudomonas and Bacillus sp. showed significant antimicrobial activities against most of the microorganisms tested. On the basis of in vitro antagonistic activities, the best strains were selected and used in field trial to study the influence of these strains on the growth of carnation. Results have shown marked effect on growth parameters and disease incidence has also been reduced significantly.

  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. Permissiveness of freshly isolated environmental strains of amoebae for growth of Legionella pneumophila.

    Science.gov (United States)

    Dupuy, Mathieu; Binet, Marie; Bouteleux, Celine; Herbelin, Pascaline; Soreau, Sylvie; Héchard, Yann

    2016-03-01

    Legionella pneumophila is a pathogenic bacterium commonly found in water and responsible for severe pneumonia. Free-living amoebae are protozoa also found in water, which feed on bacteria by phagocytosis. Under favorable conditions, some L. pneumophila are able to resist phagocytic digestion and even multiply within amoebae. However, it is not clear whether L. pneumophila could infect at a same rate a large range of amoebae or if there is some selectivity towards specific amoebal genera or strains. Also, most studies have been performed using collection strains and not with freshly isolated strains. In our study, we assess the permissiveness of freshly isolated environmental strains of amoebae, belonging to three common genera (i.e. Acanthamoeba, Naegleria and Vermamoeba), for growth of L. pneumophila at three different temperatures. Our results indicated that all the tested strains of amoebae were permissive to L. pneumophila Lens and that there was no significant difference between the strains. Intracellular proliferation was more efficient at a temperature of 40°C. In conclusion, our work suggests that, under favorable conditions, virulent strains of L. pneumophila could equally infect a large number of isolates of common freshwater amoeba genera.

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

    Directory of Open Access Journals (Sweden)

    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.

  17. Study of Corynebacterium diphtheriae strains isolated in Romania, northwestern Russia and the Republic of Moldova.

    Science.gov (United States)

    Damian, Maria; Grimont, Francine; Narvskaya, Olga; Straut, Monica; Surdeanu, Maria; Cojocaru, Radu; Mokrousov, Igor; Diaconescu, Angela; Andronescu, Constantin; Melnic, Anatol; Mutoi, Ludmila; Grimont, Patrick A D

    2002-03-01

    A selection of 167 Corynebacterium diphtheriae strains isolated in Romania, the Russian Federation and the Republic of Moldova were analysed by biotyping, phage typing, the toxin production test and by molecular techniques such as ribotyping, pulsed field gel electrophoresis and random amplified polymorphic DNA, in order to establish the epidemiological relatedness, genetic divergence and strain circulation within and between the bordering countries. Using a set of five digoxigenin-labeled oligonucleotides and BstEII digestion, 34 ribotypes were identified. The strains isolated in the epidemic areas (Russia and Moldova) were very closely related but different from those isolated in Romania. C1 and C5 were the main ribotypes identified in these areas. Neither ribotype was found in Romania, where the main circulating types were C3 and C7. Field inversion gel electrophoresis was more discriminative than ribotyping and revealed 54 macrorestriction profiles after SfiI restriction. Both methods showed a significant homogeneity of the strains from epidemic areas and a large diversity among the Romanian strains. Random amplification was useful as an identification method for the epidemic strains, but not for the Romanian ones which displayed a large number of amplification profiles. The phenotypic methods associated with molecular typing techniques enabled distinguishing between strains, detecting the epidemic clone, and sustaining the absence of transmission across borders.

  18. 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.

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

    Directory of Open Access Journals (Sweden)

    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. 

  20. Isolation and characterization of a genotype 4 Hepatitis E virus strain from an infant in China

    Directory of Open Access Journals (Sweden)

    Cui Li

    2009-02-01

    Full Text Available Abstract In the present study, a genotype 4 HEV strain was identified in the fecal specimen from a seven months old infant with no symptom of hepatitis in Shanghai Children's hospital. The full capsid protein gene (ORF2 sequence of this strain was determined by RT-PCR method. Sequence analysis based on the full ORF2 sequence indicated that this HEV strain shared the highest sequence identity (97.6% with another human HEV strain isolated from a Japanese patient who was infected by genotype 4 HEV during traveling in Shanghai. Phylogenetic analysis showed that this genotype 4 HEV was phylogenetically far from the genotype 4 HEV strain that was commonly prevalent in Shanghai swine group, suggesting that this strain may not come from swine group and not involved in zoonotic transmission in this area.

  1. Draft Genome Sequences of 40 Salmonella enterica Serovar Typhimurium Strains Isolated from Humans and Food in Brazil

    Science.gov (United States)

    Almeida, Fernanda; Medeiros, Marta Inês Cazentini; Rodrigues, Dália Prazeres; Payne, Justin; Timme, Ruth E.

    2016-01-01

    Salmonellosis is an important health problem worldwide and Salmonella enterica serovar Typhimurium is one of the most common isolated serovars. Here, we reported the draft genomes of 40 S. Typhimurium strains isolated from humans and food in Brazil. These draft genomes will improve phylogenetic analysis and will help enhance our understanding of strains of this serovar isolated in Brazil. PMID:27660768

  2. Mechanisms of food processing and storage-related stress tolerance in Clostridium botulinum.

    Science.gov (United States)

    Dahlsten, Elias; Lindström, Miia; Korkeala, Hannu

    2015-05-01

    Vegetative cultures of Clostridium botulinum produce the extremely potent botulinum neurotoxin, and may jeopardize the safety of foods unless sufficient measures to prevent growth are applied. Minimal food processing relies on combinations of mild treatments, primarily to avoid deterioration of the sensory qualities of the food. Tolerance of C. botulinum to minimal food processing is well characterized. However, data on effects of successive treatments on robustness towards further processing is lacking. Developments in genetic manipulation tools and the availability of annotated genomes have allowed identification of genetic mechanisms involved in stress tolerance of C. botulinum. Most studies focused on low temperature, and the importance of various regulatory mechanisms in cold tolerance of C. botulinum has been demonstrated. Furthermore, novel roles in cold tolerance were shown for metabolic pathways under the control of these regulators. A role for secondary oxidative stress in tolerance to extreme temperatures has been proposed. Additionally, genetic mechanisms related to tolerance to heat, low pH, and high salinity have been characterized. Data on genetic stress-related mechanisms of psychrotrophic Group II C. botulinum strains are scarce; these mechanisms are of interest for food safety research and should thus be investigated. This minireview encompasses the importance of C. botulinum as a food safety hazard and its central physiological characteristics related to food-processing and storage-related stress. Special attention is given to recent findings considering genetic mechanisms C. botulinum utilizes in detecting and countering these adverse conditions.

  3. 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.

  4. 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.

  5. [Antimicrobial resistance and molecular epidemiology of Staphylococcus pseudintermedius strains isolated from dog clinical samples].

    Science.gov (United States)

    Vigo, Germán B; Giacoboni, Gabriela I; Gagetti, Paula S; Pasterán, Fernando G; Corso, Alejandra C

    2015-01-01

    Twenty-eight strains isolated from dog clinical samples identified as Staphylococcus pseudintermedius by mass spectrometry (MALDI-TOF) were studied to assess antimicrobial susceptibility by the diffusion method and clonal relationship by pulsed field gel electrophoresis (PFGE). Methicillin resistance (3/28 isolates; 10,7%) was evaluated by mecA PCR. Fifteen strains (53.6%) were resistant to at least one of the antibiotics tested, and eleven of them (39.3%) showed multiple resistance (3 or more antimicrobial families). Eleven isolates (39.3%) were resistant to erythromycin due to the presence of ribosomal methylase ermB, whereas clindamycin inducible resistance was not detected. Twenty-seven (27) clonal types were differentiated by PFGE, suggesting high clonal diversity. We emphasize that the finding of multiresistant S. psedintermedius strains is an emerging problem to be considered in veterinary diagnostic laboratory treatment of canine infections and in public health settings.

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

    Science.gov (United States)

    Beato, Felipe B; Bergdahl, Basti; Rosa, Carlos A; Forster, Jochen; Gombert, Andreas K

    2016-11-01

    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 no clear correlation could be found either between phenotype and isolation spot or between phenotype and genomic lineage, a set of indigenous strains with superior industrially relevant traits over commonly known industrial and laboratory strains was identified: strain UFMG-CM-Y257 has a very high specific growth rate on sucrose (0.57 ± 0.02 h(-1)), high ethanol yield (1.65 ± 0.02 mol ethanol mol hexose equivalent(-1)), high ethanol productivity (0.19 ± 0.00 mol L(-1) h(-1)), high tolerance to acetic acid (10 g L(-1)) and to high temperature (40°C). Strain UFMG-CM-Y260 displayed high ethanol yield (1.67 ± 0.13 mol ethanol mol hexose equivalent(-1)), high tolerance to ethanol and to low pH, a trait which is important for non-aseptic industrial processes. Strain UFMG-CM-Y267 showed high tolerance to acetic acid and to high temperature (40°C), which is of particular interest to second generation industrial processes.

  7. 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.

  8. Isolation of bacterial strains from bovine fecal microflora capable of degradation of ceftiofur.

    Science.gov (United States)

    Rafii, Fatemeh; Williams, Anna J; Park, Miseon; Sims, Lillie M; Heinze, Thomas M; Cerniglia, Carl E; Sutherland, John B

    2009-10-20

    Ceftiofur, a third-generation cephalosporin used to treat bacterial infections in animals, is degraded in bovine feces but the specific bacteria involved are unknown. To find the bacteria involved in ceftiofur metabolism, the bovine fecal microflora was screened. Twenty-one nonidentical strains of bovine fecal bacteria were isolated on media containing 1-32 microg ml(-1) of ceftiofur. The cultures were incubated with 5 microg ml(-1) ceftiofur for different times, then centrifuged and analyzed by high-performance liquid chromatography. Three strains of Bacillus spp., two strains of Roseomonas spp., and one strain of Azospirillum sp. metabolized 5 microg ml(-1) ceftiofur in broth cultures in less than 24h; ten other strains of Roseomonas and one strain of Bacillus pumilus had metabolized it by 120 h. After the ceftiofur had been metabolized by these bacteria, the filter-sterilized supernatants of centrifuged cultures no longer inhibited the growth of a ceftiofur-sensitive strain of Kocuria rhizophila, which indicated that ceftiofur had been transformed to compounds without bactericidal activity. Each isolate was also found to be able to grow in the presence of other beta-lactams, and a nitrocefin assay showed beta-lactamase activity in the 17 strains that metabolized ceftiofur. The results show that some beta-lactamase-producing bacteria from the bovine fecal microflora are capable of transforming ceftiofur to metabolites lacking bactericidal activity.

  9. Genetic and antigenic analysis of Chlamydia pecorum strains isolated from calves with diarrhea.

    Science.gov (United States)

    Ohtani, Akifumi; Kubo, Masahito; Shimoda, Hiroshi; Ohya, Kenji; Iribe, Tadashi; Ohishi, Daiki; Endoh, Daiji; Omatsu, Tsutomu; Mizutani, Tetsuya; Fukushi, Hideto; Maeda, Ken

    2015-07-01

    Chlamydia pecorum (designated 22-58) was isolated in 2010 in HmLu-1 cells from the jejunum of a calf which died of necrotizing enterocolitis in Yamaguchi Prefecture, Japan. Immunohistochemical staining identified C. pecorum positive reactions in the jejunal villi. C. pecorum, designated 24-100, was isolated from the feces of a calf with diarrhea in another farm in Yamaguchi Prefecture in 2012. A significant increase in neutralizing antibody titers against C. pecorum was confirmed in paired sera. Nucleotide sequence identities of omp1 genes of the 2 isolates were 100%. The isolates were genetically and antigenically more closely related to C. pecorum Bo/Yokohama strain isolated from cattle with enteritis in Japan than to the other prototype strains, Bo/Maeda isolated from cattle with pneumonia and Ov/IPA isolated from sheep with polyarthritis. These results indicate that C. pecorum strains similar to 22-58 and 24-100 might be endemic in Yamaguchi Prefecture and cause enteric disease in cattle.

  10. Draft genome sequence of Thermoactinomyces sp. strain AS95 isolated from a Sebkha in Thamelaht, Algeria.

    Science.gov (United States)

    Bezuidt, Oliver K I; Gomri, Mohamed A; Pierneef, Rian; Van Goethem, Marc W; Kharroub, Karima; Cowan, Don A; Makhalanyane, Thulani P

    2016-01-01

    The members of the genus Thermoactinomyces are known for their protein degradative capacities. Thermoactinomyces sp. strain AS95 is a Gram-positive filamentous bacterium, isolated from moderately saline water in the Thamelaht region of Algeria. This isolate is a thermophilic aerobic bacterium with the capacity to produce extracellular proteolytic enzymes. This strain exhibits up to 99 % similarity with members of the genus Thermoactinomyces, based on 16S rRNA gene sequence similarity. Here we report on the phenotypic features of Thermoactinomyces sp. strain AS95 together with the draft genome sequence and its annotation. The genome of this strain is 2,558,690 bp in length (one chromosome, but no plasmid) with an average G + C content of 47.95 %, and contains 2550 protein-coding and 60 RNA genes together with 64 ORFs annotated as proteases.

  11. Isolation and characterization of tyramine-producing Enterococcus faecium strains from red wine.

    Science.gov (United States)

    Capozzi, Vittorio; Ladero, Victor; Beneduce, Luciano; Fernández, María; Alvarez, Miguel A; Benoit, Bach; Laurent, Barnavon; Grieco, Francesco; Spano, Giuseppe

    2011-05-01

    Enterococcus faecium strains were isolated from red wines undergoing malolactic fermentation and identified by comparison of their 16S rDNA gene sequences with those included in the GenEMBL Databases. The tyrosine decarboxylase gene was identified in all the strains analysed by PCR using gene-specific primers and the ability to produce tyramine in a synthetic media was analysed by RP-HPLC. Survival of an E. faecium strain was also evaluated in microvinification assays using two different musts with different ethanol concentrations (10% and 12% (v/v)). Tyramine production was monitored during the vinification trials. Our results suggest that E. faecium strains isolated from wine are able to produce tyramine and tolerate wine conditions following a pre-acidic stress.

  12. [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.

  13. Draft genome sequences for two metal-reducing Pelosinus fermentans strains isolated from a Cr(VI)-contaminated site and for type strain R7.

    Science.gov (United States)

    Brown, Steven D; Podar, Mircea; Klingeman, Dawn M; Johnson, Courtney M; Yang, Zamin K; Utturkar, Sagar M; Land, Miriam L; Mosher, Jennifer J; Hurt, Richard A; Phelps, Tommy J; Palumbo, Anthony V; Arkin, Adam P; Hazen, Terry C; Elias, Dwayne A

    2012-09-01

    Pelosinus fermentans 16S rRNA gene sequences have been reported from diverse geographical sites since the recent isolation of the type strain. We present the genome sequence of the P. fermentans type strain R7 (DSM 17108) and genome sequences for two new strains with different abilities to reduce iron, chromate, and uranium.

  14. Dental caries induction in experimental animals by clinical strains of Streptococcus mutans isolated from Japanese children.

    Science.gov (United States)

    Hamada, S; Ooshima, T; Torii, M; Imanishi, H; Masuda, N; Sobue, S; Kotani, S

    1978-01-01

    Oral implantation and the cariogenic activity of clinical strains of Streptococcus mutans which had been isolated from Japanese children and labeled with streptomycin-resistance were examined in specific pathogen-free Sprague-Dawley rats. All the seven strains tested were easily implanted and persisted during the experimental period. Extensive carious lesions were produced in rats inoculated with clinical strains of S. mutans belonging to serotypes c, d, e, and f, and maintained on caries-inducing diet no. 2000. Noninfected rats did not develop dental caries when fed diet no. 2000. Type d S. mutans preferentially induced smooth surface caries in the rats. Strains of other serotypes primarily developed caries of pit and fissure origin. Caries also developed in rats inoculated with reference S. mutans strains BHTR and FAIR (type b) that had been maintained in the laboratories for many years. However, the cariogenicity of the laboratory strains was found to have decreased markedly. All three S. sanguis strains could be implanted, but only one strain induced definite fissure caries. Two S. salivarius strains could not be implanted well in the rats and therefore they were not cariogenic. Four different species of lactobacilli also failed to induce dental caries in rats subjected to similar caries test regimen on diet no. 200. S. mutans strain MT6R (type c) also induce caries in golden hamsters and ICR mice, but of variable degrees.

  15. Effect of CuO Nanoparticles over Isolated Bacterial Strains from Agricultural Soil

    Directory of Open Access Journals (Sweden)

    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.

  16. Comparative analysis of tertiary alcohol esterase activity in bacterial strains isolated from enrichment cultures and from screening strain libraries.

    Science.gov (United States)

    Herter, Susanne; Nguyen, Giang-Son; Thompson, Mark L; Steffen-Munsberg, Fabian; Schauer, Frieder; Bornscheuer, Uwe T; Kourist, Robert

    2011-05-01

    The preparation of enantiopure tertiary alcohols is of great contemporary interest due to the application of these versatile building blocks in organic synthesis and as precursors towards high value pharmaceutical compounds. Herein, we describe two approaches taken towards the discovery of novel biocatalysts for the synthesis of these valuable compounds. The first approach was initiated with screening of 47 bacterial strains for hydrolytic activity towards the simple tertiary alcohol ester tert-butyl acetate. In conjunction, a second method focussed on the isolation of strains competent for growth on tert-butyl acetate as the sole source of carbon and energy. From functional screening, 10 Gram-positive Actinomycetes showed hydrolytic activity, whilst enrichment selection resulted in the identification of 14 active strains, of which five belong to the Gram-negative cell-wall type. Bacterial strains obtained from both approaches were viable for enantioselective hydrolysis of pyridine substituted tertiary alcohol esters in addition to bulky aliphatic and keto-derived substrates from the same class. Activity towards each of the test substrates was uncovered, with promising enantioselectivities of up to E = 71 in the hydrolysis of a para-substituted pyridine tertiary alcohol ester using a strain of Rhodococcus ruber. Interestingly strains of Microbacterium and Alcaligenes sp. gave opposite enantiopreference in the hydrolysis of a meta-substituted pyridine tertiary alcohol ester with E values of 17 and 54. These approaches show that via both possibilities, screening established strain collections and performing enrichment selection, it is possible to identify novel species which show activity towards sterically challenging substrates.

  17. Draft Genome Sequence of Enterococcus faecalis Strain F165 Isolated from a Urinary Tract Infection

    Science.gov (United States)

    Pieta, Luiza; Prichula, Janira; Sambrano, Gustavo E.; Soares, Renata; Bello, Aline Dall; Frazzon, Jeverson; d’Azevedo, Pedro A.

    2016-01-01

    We report here a draft genome sequence of Enterococcus faecalis strain F165 isolated from a urine specimen in South Brazil. The genome size was 3,049,734 bp, with a G+C content of 37.38%, and genes related to antimicrobial resistance and adherence were found in the strain. These findings are consistent with pathogenesis of E. faecalis species.

  18. Draft Genome Sequence of Burkholderia cordobensis Type Strain LMG 27620, Isolated from Agricultural Soils in Argentina

    Science.gov (United States)

    Draghi, Walter Omar; Mancini Villagra, Ulises M.; Wall, Luis Gabriel

    2015-01-01

    Bacteria of the genus Burkholderia are commonly found in diverse ecological niches in nature. We report here the draft genome sequence of Burkholderia cordobensis type strain LMG 27620, isolated from agricultural soil in Córdoba, Argentina. This strain harbors several genes involved in chitin utilization and phenol degradation, which make it an interesting candidate for biocontrol purposes and xenobiotic degradation in polluted environments. PMID:26494680

  19. Genome Sequences of 14 Firmicutes Strains Isolated from the Human Vagina

    Science.gov (United States)

    Deitzler, Grace E.; Ruiz, Maria J.; Weimer, Cory; Park, SoEun; Robinson, Lloyd; Hallsworth-Pepin, Kymberlie; Wollam, Aye; Mitreva, Makedonka

    2016-01-01

    Research on vaginal infections is currently limited by a lack of available fully sequenced bacterial reference strains. Here, we present strains (now available through BEI Resources) and genome sequences for a set of 14 vaginal isolates from the phylum Firmicutes. These genome sequences provide a valuable resource for future research in understanding the role of Gram-positive bacteria in vaginal health and disease. PMID:27688329

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

    OpenAIRE

    2015-01-01

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

  1. 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 sugarcan

  2. Genome Sequence of Staphylococcus pseudintermedius Strain E140, an ST71 European-Associated Methicillin-Resistant Isolate.

    Science.gov (United States)

    Moodley, Arshnee; Riley, Matthew C; Kania, Stephen A; Guardabassi, Luca

    2013-03-07

    We report the first genome sequence of the methicillin-resistant Staphylococcus pseudintermedius (MRSP) strain E140, isolated from a canine bite wound infection in Denmark. This strain represents the dominant clonal lineage associated with canine MRSP infections in Europe.

  3. Antibiotic Resistance of Probiotic Strains of Lactic Acid Bacteria Isolated from Marketed Foods and Drugs

    Institute of Scientific and Technical Information of China (English)

    CHANG LIU; ZHUO-YANG ZHANG; KE DONG; JIAN-PING YUAN; XIAO-KUI GUO

    2009-01-01

    Objective To identify the antimicrobial resistance of commercial lactic acid bacteria present in microbial foods and drug additives by analyzing their isolated strains used for fermentation and probioties. Methods Antimicrobial susceptibility of 41 screened isolates was tested with disc diffusion and E-test methods after species-level identification. Resistant strains were selected and examined for the presence of resistance genes by PCR. Results Distribution of resistance was found in different species. All isolates were susceptible to chloramphenicol, tetracycline, ampicillin, amoxicillin/clavulanic acid, cephalothin, and imipenem. In addition, isolates resistant to vancomycin, rifampicin, streptomycin, bacitracin, and erythromycin were detected, although the incidence of resistance to these antibiotics was relatively low. In contrast, most strains were resistant to ciprofloxacin, amikacin, trimethoprim/sulphamethoxazole, and gentamycin. The genes msrC, vanX, and dfrA were detected in strains of Enterococcus faecium, Lactobacillus plantarum, Streptococcus thermophilus, and Lactococcus lactis. Conclusion Antibiotic resistance is present in different species of probiotic strains, which poses a threat to food safety. Evaluation of the safety of lactic acid bacteria for human consumption should be guided by established criteria, guidelines and regulations.

  4. Isolation and selection of new astaxanthin producing strains of Xanthophyllomyces dendrorhous.

    Science.gov (United States)

    Libkind, Diego; Moliné, Martín; Tognetti, Celia

    2012-01-01

    Astaxanthin is a xanthophyll pigment of high economic value for its use as a feeding component in aquaculture. Xanthophyllomyces dendrorhous is a basidiomycetous fungi able to synthesize astaxanthin as its major carotenoid, and the only known yeast species bearing the capability to produce this type of carotenoid. Recently, the habitat and intraspecific variability of this species have been found to be wider than previously expected, encouraging the search for new wild strains with potential biotechnological applications. Here we describe effective procedures for isolation of X. dendrorhous from environmental samples, accurate identification of the strains, analysis of their astaxanthin content, and proper conservation of the isolates.

  5. Features of Lactobacillus sakei isolated from Italian sausages: focus on strains from Ventricina del Vastese

    Directory of Open Access Journals (Sweden)

    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.

  6. Comparison of the serum sensitivity of uropathogenic strains of Escherichia coli isolated from different diagnostic groups

    Directory of Open Access Journals (Sweden)

    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.

  7. Isolation of Escherichia coli 0157:H7 Strain from Fecal Samples of Zoo Animal

    Directory of Open Access Journals (Sweden)

    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.

  8. Isolation of Escherichia coli 0157:H7 strain from fecal samples of zoo animal.

    Science.gov (United States)

    Mohammed Hamzah, Aseel; Mohammed Hussein, Aseel; Mahmoud Khalef, Jenan

    2013-01-01

    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.

  9. 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.

  10. "Emergence of Multidrug Resistant Strains of Escherichia coli Isolated from Urinary Tract Infections"

    OpenAIRE

    R Moniri; Khorshidi, A; H Akbari

    2003-01-01

    The emergence of multidrug resistant strains of Escherichia coli has complicated treatment decision and may lead to treatment failures. From April to November 2001 we prospectively evaluated the prevalence of resistance to trimethoprim-sulfamethoxazole (SXT), gentamicin, cephalothin, ciprofloxacin, and nitrofurantoin in 220 Escherichia coli isolates from patients with urinary tract infections in kashan, Iran. To assess the current breadth of multidrug resistance among urinary isolates of E. c...

  11. Draft genome sequences of four Achromobacter ruhlandii strains isolated from cystic fibrosis patients

    Science.gov (United States)

    Rodrigues, Elenice RA; Rocha, Géssica A; Ferreira, Alex G; Leão, Robson S; Albano, Rodolpho M; Marques, Elizabeth A

    2016-01-01

    Achromobacter species are being increasingly isolated from the respiratory tract of cystic fibrosis patients. Recent reports indicate that Achromobacter ruhlandii is a potential human pathogen in cystic fibrosis-related infections. Here we report the draft genome of four A. ruhlandii strains isolated from cystic fibrosis patients in Brazil. This report describes A. ruhlandii as a potential opportunistic pathogen in cystic fibrosis and provides a framework to for additional enquires into potential virulence factors and resistance mechanisms within this species. PMID:27812598

  12. Isolation and Identification of Oedogonium Species and Strains for Biomass Applications

    OpenAIRE

    Rebecca J Lawton; Rocky de Nys; Stephen Skinner; Nicholas A Paul

    2014-01-01

    Freshwater macroalgae from the genus Oedogonium have recently been targeted for biomass applications; however, strains of Oedogonium for domestication have not yet been identified. Therefore, the objective of this study was to compare the performance of isolates of Oedogonium collected from multiple geographic locations under varying environmental conditions. We collected and identified wild-type isolates of Oedogonium from three geographic locations in Eastern Australia, then measured the gr...

  13. 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.

  14. Morphological, physiological and plant infectivity characterization of Frankia strains isolated from Casuarina’s nodules

    OpenAIRE

    Patrick Moritz; Kelly Campos Guerra P. de Goes; Souza,José Roberto P de; Letícia Trindade Ataíde; Diva Souza Andrade

    2007-01-01

    Frankia are soil microorganisms that form symbiosis with roots of tree species called actinorhizal plants and are capable of fixing atmospheric N2. This study was carried out to characterize morphologically, physiologically and to assess the nodulation of four Frankia reference strains (HFPCcI3, JCT287, KB5 and F59) and 12 (IPRF) isolated from root nodules of Casuarina plants. All strains (Reference and IPRF) were characterized as Gram-positive and 50% as acid-fast. The Frankia strains produc...

  15. Isolation, identification and characterization of an electrogenic microalgae strain.

    Science.gov (United States)

    Wu, Yicheng; Guan, Kai; Wang, Zejie; Xu, Bing; Zhao, Feng

    2013-01-01

    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.

  16. 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.

  17. Large scale analysis of virulence genes in Escherichia coli strains isolated from Avalon Bay, CA.

    Science.gov (United States)

    Hamilton, Matthew J; Hadi, Asbah Z; Griffith, John F; Ishii, Satoshi; Sadowsky, Michael J

    2010-10-01

    Contamination of recreational waters with Escherichia coli and Enterococcus sp. is a widespread problem resulting in beach closures and loss of recreational activity. While E. coli is frequently used as an indicator of fecal contamination, and has been extensively measured in waterways, few studies have examined the presence of potentially pathogenic E. coli strains in beach waters. In this study, a combination of high-throughput, robot-assisted colony hybridization and PCR-based analyses were used to determine the genomic composition and frequency of virulence genes present in E. coli isolated from beach water in Avalon Bay, Santa Catalina Island, CA. A total of 24,493 E. coli isolates were collected from two sites at a popular swimming beach between August through September 2007 and from July through August 2008. All isolates were examined for the presence of shiga-like toxins (stx1/stx2), intimin (eaeA), and enterotoxins (ST/LT). Of the 24,493 isolates examined, 3.6% contained the eaeA gene, indicating that these isolates were potential EPEC strains. On five dates, however, greater than 10% of the strains were potential EPEC, suggesting that incidence of virulence genes at this beach has a strong temporal component. No STEC or ETEC isolates were detected, and only eight (water and their presence needs to be considered as one of the factors used in decisions concerning beach closures.

  18. Isolation and characterization of different strains of Bacillus licheniformis for the production of commercially significant enzymes.

    Science.gov (United States)

    Ghani, Maria; Ansari, Asma; Aman, Afsheen; Zohra, Rashida Rahmat; Siddiqui, Nadir Naveed; Qader, Shah Ali Ul

    2013-07-01

    Utilization of highly specific enzymes for various industrial processes and applications has gained huge momentum in the field of white biotechnology. Selection of a strain by efficient plate-screening method for a specific purpose has also favored and boosted the isolation of several industrially feasible microorganisms and screening of a large number of microorganisms is an important step in selecting a potent culture for multipurpose usage. Five new bacterial isolates of Bacillus licheniformis were discovered from indigenous sources and characterized on the basis of phylogeny using 16S rDNA gene analysis. Studies on morphological and physiological characteristics showed that these isolates can easily be cultivated at different temperatures ranging from 30°C to 55°C with a wide pH values from 3.0 to 11.0 All these 05 isolates are salt tolerant and can grow even in the presences of high salt concentration ranging from 7.0 to 12.0%. All these predominant isolates of B. licheniformis strains showed significant capability of producing some of the major industrially important extracellular hydrolytic enzymes including α-amylase, glucoamylase, protease, pectinase and cellulase in varying titers. All these isolates hold great potential as commercial strains when provided with optimum fermentation conditions.

  19. Investigation of the Antimicrobial Activity of Bacillus licheniformis Strains Isolated from Retail Powdered Infant Milk Formulae.

    Science.gov (United States)

    Alvarez-Ordóñez, Avelino; Begley, Máire; Clifford, Tanya; Deasy, Thérèse; Considine, Kiera; O'Connor, Paula; Ross, R Paul; Hill, Colin

    2014-03-01

    This study investigated the potential antimicrobial activity of ten Bacillus licheniformis strains isolated from retail infant milk formulae against a range of indicator (Lactococcus lactis, Lactobacillus bulgaricus and Listeria innocua) and clinically relevant (Listeria monocytogenes, Staphylococcus aureus, Streptococcus agalactiae, Salmonella Typhimurium and Escherichia coli) microorganisms. Deferred antagonism assays confirmed that all B. licheniformis isolates show antimicrobial activity against the Gram-positive target organisms. PCR and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analyses indicated that four of the B. licheniformis isolates produce the bacteriocin lichenicidin. The remaining six isolates demonstrated a higher antimicrobial potency than lichenicidin-producing strains. Further analyses identified a peptide of ~1,422 Da as the most likely bioactive responsible for the antibacterial activity of these six isolates. N-terminal sequencing of the ~1,422 Da peptide from one strain identified it as ILPEITXIFHD. This peptide shows a high homology to the non-ribosomal peptides bacitracin and subpeptin, known to be produced by Bacillus spp. Subsequent PCR analyses demonstrated that the six B. licheniformis isolates may harbor the genetic machinery needed for the synthesis of a non-ribosomal peptide synthetase similar to those involved in production of subpeptin and bacitracin, which suggests that the ~1,422 Da peptide might be a variant of subpeptin and bacitracin.

  20. Biodegradation of Chlorpyrifos by Pseudomonas Resinovarans Strain AST2.2 Isolated from Enriched Cultures.

    Directory of Open Access Journals (Sweden)

    Anish Sharma*,

    2016-04-01

    Full Text Available A bacterial strain AST2.2 with chlorpyrifos degrading ability was isolated by enrichment technique from apple orchard soil with previous history of chlorpyrifos use. Based on the morphological, biochemical tests and 16S rRNA sequence analysis, AST2.2 strain was identified as Pseudomonas resinovarans. The strain AST2.2 utilized chlorpyrifos as the sole source of carbon and energy. This strain exhibited growth upto 400mg/l concentration of chlorpyrifos and exhibited high extracellular organophosphorus hydrolase (OPH activity. Gas chromatography-flame ionization detector (GC-FID studies revealed that Pseudomonas resinovarans AST2.2 degraded 43.90 % of chlorpyrifos (400 mg/l within 96 hrs. Intermediates of chlorpyrifos degradation were identified using GC-MS. This strain have potential to degrade chlorpyrifos and thus can be used for bioremediation and ecological restoration of sites contaminated with chlorpyrifos.

  1. 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.

  2. Draft Genome Sequences of Tersicoccus phoenicis DSM 30849T, Isolated from a Cleanroom for Spacecraft Assembly, and Tersicoccus sp. Strain Bi-70, Isolated from a Freshwater Lake

    Science.gov (United States)

    Yoshizawa, Susumu; Nakamura, Keiji; Ogura, Yoshitoshi; Hayashi, Tetsuya; Kogure, Kazuhiro

    2017-01-01

    ABSTRACT Here, we report the draft genome sequences of Tersicoccus phoenicis DSM 30849T, isolated from a spacecraft assembly cleanroom at the National Aeronautics and Space Administration (NASA), and Tersicoccus sp. strain Bi-70, isolated from Lake Biwa, the largest lake in Japan. These genome sequences facilitate our understanding of the adaptation of these closely related strains to different habitats. PMID:28360156

  3. 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.

  4. Isolation of a strain of Pseudomonas putida capable of metabolizing anionic detergent sodium dodecyl sulfate (SDS

    Directory of Open Access Journals (Sweden)

    A Kumar

    2011-05-01

    Full Text Available Background and objectives: 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."nMaterials and Methods: 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."nResults: 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."nConclusion: 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.

  5. 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.

  6. 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.

  7. 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 c

  8. A Desulfitobacterium strain isolated from human feces that does not dechlorinate chloroethenes or chlorophenols

    NARCIS (Netherlands)

    van de Pas, BA; Harmsen, HJM; Raangs, GC; de Vos, WM; Schraa, G; Stams, AJM

    2001-01-01

    An anaerobic bacterium, strain DP7, was isolated from human feces in mineral medium with formate and 0.02% yeast extract as energy and carbon source. This rod-shaped motile bacterium used pyruvate, lactate, formate, hydrogen, butyrate, and ethanol as electron donor for sulfite reduction. Other elect

  9. Draft Genome Sequences of Seven Pseudomonas fluorescens Subclade III Strains Isolated from Cystic Fibrosis Patients.

    Science.gov (United States)

    Scales, Brittan S; Erb-Downward, John R; Huffnagle, Ian M; LiPuma, John J; Huffnagle, Gary B

    2015-01-29

    We report here the first draft genome sequences of Pseudomonas fluorescens strains that have been isolated from humans. The seven assembled draft genomes contained an average of 60.1% G+C content, were an average genomic size of 6.3 Mbp, and mapped by multilocus sequence analysis to subclade III.

  10. Draft Genome Sequence of Lactobacillus gorillae Strain KZ01T, Isolated from a Western Lowland Gorilla

    OpenAIRE

    TSUCHIDA, Sayaka; Nezuo, Maiko; Tsukahara, Masatoshi; Ogura, Yoshitoshi; Hayashi, Tetsuya; Ushida, Kazunari

    2015-01-01

    Here, we report the draft genome sequence of Lactobacillus gorillae strain KZ01T isolated from a western lowland gorilla (Gorilla gorilla gorilla). This genome sequence will be helpful for the comparative genomics between human and nonhuman primate-associated Lactobacillus.

  11. Draft Genome Sequences of Two Salmonella enterica Strains Isolated from Sprouted Chia and Flax Seed Powders

    OpenAIRE

    Ronholm, Jennifer; Petronella, Nicholas; Tamber, Sandeep

    2016-01-01

    A 2014 foodborne salmonellosis outbreak in Canada and the United States implicated, for the first time, sprouted chia seed powder as the vehicle of transmission. Here, we report the draft whole genome sequences of two Salmonella enterica strains isolated from sprouted powders related to the aforementioned outbreak.

  12. A new carotenoid glycosyl ester isolated from a marine microorganism, Fusarium strain T-1.

    Science.gov (United States)

    Sakaki, Hideyuki; Kaneno, Hirokazu; Sumiya, Yasuji; Tsushima, Miyuki; Miki, Wataru; Kishimoto, Noriaki; Fujita, Tokio; Matsumoto, Sadayoshi; Komemushi, Sadao; Sawabe, Akiyoshi

    2002-11-01

    A new carotenoid glycosyl ester, neurosporaxanthin beta-D-glucopyranoside (2), together with neurosporaxanthin (1), beta-carotene, gamma-carotene, and torulene were isolated from cultured cells of a marine microorganism, strain T-1, which was identified as Fusarium sp. Their structures were determined by chemical and spectral data.

  13. Draft Genome Sequence of Acetobacter malorum CECT 7742, a Strain Isolated from Strawberry Vinegar.

    Science.gov (United States)

    Sainz, Florencia; Mas, Albert; Torija, María Jesús

    2016-06-23

    The present article reports the draft genome sequence of the strain Acetobacter malorum CECT 7742, an acetic acid bacterium isolated from strawberry vinegar. This species is characterized by the production of d-gluconic acid from d-glucose, which it further metabolizes to keto-d-gluconic acids.

  14. Screening of bacterial strains isolated from uranium mill tailings porewaters for bioremediation purposes.

    Science.gov (United States)

    Sánchez-Castro, Iván; Amador-García, Ahinara; Moreno-Romero, Cristina; López-Fernández, Margarita; Phrommavanh, Vannapha; Nos, Jeremy; Descostes, Michael; Merroun, Mohamed L

    2017-01-01

    The present work characterizes at different levels a number of bacterial strains isolated from porewaters sampled in the vicinity of two French uranium tailing repositories. The 16S rRNA gene from 33 bacterial isolates, corresponding to the different morphotypes recovered, was almost fully sequenced. The resulting sequences belonged to 13 bacterial genera comprised in the phyla Firmicutes, Actinobacteria and Proteobacteria. Further characterization at physiological level and metals/metalloid tolerance provided evidences for an appropriate selection of bacterial strains potentially useful for immobilization of uranium and other common contaminants. By using High Resolution Transmission Electron Microscope (HRTEM), this potential ability to immobilize uranium as U phosphate mineral phases was confirmed for the bacterial strains Br3 and Br5 corresponding to Arthrobacter sp. and Microbacterium oxydans, respectively. Scanning Transmission Electron Microscope- High-Angle Annular Dark-Field (STEM-HAADF) analysis showed U accumulates on the surface and within bacterial cytoplasm, in addition to the extracellular space. Energy Dispersive X-ray (EDX) element-distribution maps demonstrated the presence of U and P within these accumulates. These results indicate the potential of certain bacterial strains isolated from porewaters of U mill tailings for immobilizing uranium, likely as uranium phosphates. Some of these bacterial isolates might be considered as promising candidates in the design of uranium bioremediation strategies.

  15. Genome Sequences of Two Leuconostoc pseudomesenteroides Strains Isolated from Danish Dairy Starter Cultures

    DEFF Research Database (Denmark)

    Pedersen, T.B.; Kot, Witold; Hansen, Lars Hestbjerg;

    2014-01-01

    The lactic acid bacterium Leuconostoc pseudomesenteroides can be found in mesophilic cheese starters, where it produces aromatic compounds from, e.g., citrate. Here, we present the draft genome sequences of two L. pseudomesenteroides strains isolated from traditional Danish cheese starters....

  16. Genome Sequences of Two Leuconostoc pseudomesenteroides Strains Isolated from Danish Dairy Starter Cultures

    Science.gov (United States)

    Kot, W. P.; Hansen, L. H.; Sørensen, S. J.; Broadbent, J. R.; Vogensen, F. K.; Ardö, Y.

    2014-01-01

    The lactic acid bacterium Leuconostoc pseudomesenteroides can be found in mesophilic cheese starters, where it produces aromatic compounds from, e.g., citrate. Here, we present the draft genome sequences of two L. pseudomesenteroides strains isolated from traditional Danish cheese starters. PMID:24903866

  17. Complete Genome Sequence of Nonagglutinating Lactococcus garvieae Strain 122061 Isolated from Yellowtail in Japan

    Science.gov (United States)

    Nishiki, Issei; Oinaka, Daisaku; Iwasaki, Yuki; Yasuike, Motoshige; Nakamura, Yoji; Yoshida, Terutoyo; Nagai, Satoshi; Katoh, Masaya; Kobayashi, Takanori

    2016-01-01

    Nonagglutinating Lactococcus garvieae has been isolated from diseased farmed yellowtail in Japan since 2012. In this study, the complete genome and plasmid sequence of nonagglutinating L. garvieae strain 122061 was determined, to our knowledge, for the first time. PMID:27389264

  18. Induction, isolation, and characterization of aspergillus niger mutant strains producing elevated levels of beta-galactosidase.

    OpenAIRE

    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.

  19. Genome sequences of two Leuconostoc pseudomesenteroides strains isolated from Danish dairy starter cultures

    DEFF Research Database (Denmark)

    Pedersen, Thomas Bæk; Kot, Witold Piotr; Hansen, L.H.;

    2014-01-01

    The lactic acid bacterium Leuconostoc pseudomesenteroides can be found in mesophilic cheese starters, where it produces aromatic compounds from, e.g., citrate. Here, we present the draft genome sequences of two L. pseudomesenteroides strains isolated from traditional Danish cheese starters....

  20. Genome Sequence of Lactobacillus brevis Strain D6, Isolated from Smoked Fresh Cheese.

    Science.gov (United States)

    Kant, Ravi; Uroić, Ksenija; Hynönen, Ulla; Kos, Blaženka; Šušković, Jagoda; Palva, Airi

    2016-04-07

    The autochthonousLactobacillus brevisstrain D6, isolated from smoked fresh cheese, carries a 45-kDa S-layer protein. Strain D6 has shown adhesion to extracellular matrix proteins and to Caco-2 intestinal epithelial cells, as well as immunomodulatory potential and beneficial milk technological properties. Hence, it could be used as a potential probiotic starter culture for cheese production.

  1. 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...

  2. 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.

  3. A Serotype VIII Strain among Colonizing Group B Streptococcal Isolates in Boston, Massachusetts

    OpenAIRE

    Paoletti, Leanne J.; Bradford, Jessica; Paoletti, Lawrence C.

    1999-01-01

    Maternal colonization with group B Streptococcus (GBS) is a risk factor for neonatal GBS disease. Whereas serotypes Ia, Ib, II, III, and V are prevalent in the United States, types VI and VIII predominate in Japan. Recently, a serotype VIII strain was detected among 114 clinical GBS isolates from a Boston, Mass., hospital.

  4. Draft Genome Sequence of Phenylobacterium immobile Strain E (DSM 1986), Isolated from Uncontaminated Soil in Ecuador

    OpenAIRE

    Reznicek, Ondrej; Luesken, Francisca; Facey, Sandra J.; Hauer, Bernhard

    2015-01-01

    We report the draft genome sequence of 3.3 Mb and the sequence (19.2 kb) of a natural plasmid isolated from Phenylobacterium immobile strain E (DSM 1986), able to degrade xenobiotic compounds as the sole carbon source. The sequences reveal a large number of novel Rieske nonheme iron aromatic ring-hydroxylating oxygenases (RHOs).

  5. Complete Genome Sequence of Streptococcus iniae 89353, a Virulent Strain Isolated from Diseased Tilapia in Taiwan

    Science.gov (United States)

    Wu, Sheng-Han; Chen, Chun-Yao; Huang, Chang-Wen; Lu, Jenn-Kan; Chou, Hsin-Yiu

    2017-01-01

    ABSTRACT Streptococcus iniae 89353 is a virulent strain isolated from diseased tilapia in Taiwan. The full-genome sequence of S. iniae 89353 is 2,098,647 bp. The revealed genome information will be beneficial for identification and understanding of potential virulence genes of Streptococcus iniae and possible immunogens for vaccine development against streptococcosis. PMID:28126946

  6. Draft Genome Sequences of Four Salmonella enterica Strains Isolated from Turkey-Associated Sources

    Science.gov (United States)

    Han, Jing; Gokulan, Kuppan; Zhao, Shaohua; Gies, Allen

    2016-01-01

    We report the draft genomes of four Salmonella enterica isolates evaluated for the contribution of plasmids to virulence. Strains SE163A, SE696A, and SE710A carry plasmids demonstrated to facilitate plasmid-associated virulence, while SE819 is less virulent and has been used as a recipient for conjugation experiments to assess plasmid-encoded virulence mechanisms. PMID:27738037

  7. 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...

  8. Draft Genome Sequence of Acetobacter malorum CECT 7742, a Strain Isolated from Strawberry Vinegar

    Science.gov (United States)

    Sainz, Florencia; Torija, María Jesús

    2016-01-01

    The present article reports the draft genome sequence of the strain Acetobacter malorum CECT 7742, an acetic acid bacterium isolated from strawberry vinegar. This species is characterized by the production of d-gluconic acid from d-glucose, which it further metabolizes to keto-d-gluconic acids. PMID:27340078

  9. Two Lactobacillus strains, isolated from breast milk, differently modulate the immune response

    NARCIS (Netherlands)

    Diaz-Ropero, M.P.; Martin, R.; Sierra, S.; Lara-Villoslada, F.; Rodriguez, J.M.; Xaus, J.; Olivares, M.

    2007-01-01

    Aims: The ability of two different Lactobacillus strains (Lactobacillus salivarius CECT5713 and Lactobacillus fermentum CECT5716), isolated from human breast milk, to modulate the immune response was examined. Methods and Results: In rodent bone-marrow-derived macrophages (BMDM), the presence of Lac

  10. Draft Genome Sequences of Five Clinical Strains of Brucella melitensis Isolated from Patients Residing in Kuwait

    Science.gov (United States)

    Khan, Mohd Wasif; Habibi, Nazima; Shaheed, Faraz

    2016-01-01

    Human brucellosis is a neglected and underrecognized infection of widespread geographic distribution. Brucellosis is present on all inhabited continents and endemic in many areas of the world, including Kuwait and the Middle East. Here, we present draft genome assemblies of five Brucella melitensis strains isolated from brucellosis patients in Kuwait. PMID:27811090

  11. Complete Genome Sequence of Enterococcus faecalis Strain W11 Isolated from an Algal Food Product

    Science.gov (United States)

    Takizawa, Noboru

    2016-01-01

    Here, we report the complete genome sequence of Enterococcus faecalis strain W11 isolated from an algal food product in Japan. This study should facilitate the identification of a novel mechanism of glycerol metabolic control in lactic acid bacteria. PMID:27688337

  12. 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.

  13. Antimicrobial Activity of Monoramnholipids Produced by Bacterial Strains Isolated from the Ross Sea (Antarctica) †

    Science.gov (United States)

    Tedesco, Pietro; Maida, Isabel; Palma Esposito, Fortunato; Tortorella, Emiliana; Subko, Karolina; Ezeofor, Chidinma Christiana; Zhang, Ying; Tabudravu, Jioji; Jaspars, Marcel; Fani, Renato; de Pascale, Donatella

    2016-01-01

    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. PMID:27128927

  14. 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.

  15. Plasmid Profiling and Curing of Lactobacillus Strains Isolated from Fermented Milk for Probiotic Applications

    Directory of Open Access Journals (Sweden)

    B. Lavanya

    2011-04-01

    Full Text Available In this study, the antimicrobial susceptibilities and presence of plasmids in 7 probiotics strains which had been isolated from the fermented milk were determined. Resistance to 8 commonly used antibiotics $- lactans (penicillin, ampicilin, gram positive spectrum (vanomycin, broad spectrum (rifampin, trimethoprim and aminogycosides (kanamycin, streptomycin, and bacitracin was assessed by disk diffusion. Among these strains 20, 20, 60, 70, 90 and 100% were found to be exhibit a significant degree of resistance to kanamycin, trimetroprim, rifampicin, kanamycin, amphicilin and penicillin respectively. Further, plasmid profile and curing of plasmid were performed for the seven isolates. Analysis of the plasmid profiles of the 7 cured derivatives revealed loss of plasmids except 2 strains where curing was partially effective. All the strains lost penicillin resistance after curing indicating that plasmids encodes for resistance character. However, vanomycin resistance is not lost upon curing which indicates that such resistance is usually intrinsic (chromosomally encoded and not transmissible. Finally, the antimicrobial susceptibility after curing was done to check the safety aspect of the isolates for their application as probiotics and among the 7 strains, 5 were proved to be potent probiotics.

  16. Characterization of Naegleria fowleri strains isolated from human cases of primary amoebic meningoencephalitis in Mexico.

    Science.gov (United States)

    Cervantes-Sandoval, Isaac; de Serrano-Luna, José Jesús; Tapia-Malagón, José Luis; Pacheco-Yépez, Judith; Silva-Olivares, Angélica; Galindo-Gómez, Silvia; Tsutsumi, Victor; Shibayama, Mineko

    2007-01-01

    The protozoon Naegleria fowleri (N. fowleri) is a free-living amoeba that produces primary amoebic meningoencephalitis (PAM), which is an acute and frequently fatal infection of the central nervous system. We characterized the strains of N. fowleri isolated from the cerebrospinal fluid (CSF) of two cases presented in northwestern Mexico. The strains were isolated and cultured in 2% bactocasitone medium. Enflagellation assays, ultrastructural analysis, protein and protease electrophoresis patterns, and PCR were performed as confirmatory tests. Virulence tests were done using in Balb/c mice. Light microscopy analysis of brain tissue showed amoebae with abundant inflammatory reaction and extensive necrotic and hemorrhagic areas. The enflagellation assay was positive and the electron microscopy showed trophozoites with morphologic features typical of the genus. Protein and protease profiles of the isolated strains were identical to the reference strain. Finally, a 1500-bp PCR product was found in all three strains. Based on all the analyses performed, we concluded that the etiologic agent of both PAM cases was N. fowleri. The need for better epidemiological information and educational programs about basic clinical and pathological aspects of free-living amoebae provided by the health authorities are emphasized.

  17. 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...... from the T7 RNA polymerase (P)-based BL21(DE3) protein production strain. In C41(DE3) and C43(DE3) mutations lowering t7rnap expression levels result in strongly reduced T7 RNAP accumulation levels. As a consequence membrane protein production stress is alleviated in the C41(DE3) and C43(DE3) strains......, thereby increasing membrane protein yields. Here, we isolated Mutant56(DE3) from BL21(DE3) using a genetic screen designed to isolate BL21(DE3)-derived strains with mutations alleviating membrane protein production stress other than the ones in C41(DE3) and C43(DE3). The defining mutation of Mutant56(DE3...

  18. Isolation and typification of histamine-producing Lactobacillus vaginalis strains from cheese.

    Science.gov (United States)

    Diaz, Maria; del Rio, Beatriz; Ladero, Victor; Redruello, Begoña; Fernández, María; Martin, Maria Cruz; Alvarez, Miguel A

    2015-12-23

    In food, the biogenic amine (BA) histamine is mainly produced by histidine decarboxylation catalysed by microbial histidine decarboxylase. The consumption of foods containing high concentrations of histamine can trigger adverse neurological, gastrointestinal and respiratory reactions. Indeed, histamine is one of the most toxic of all BAs, and is often detected in high concentration in cheese. However, little is known about the microorganisms responsible for its accumulation in this food. In the present work, 25 histamine-producing Lactobacillus vaginalis strains were isolated from a blue-veined cheese (the first time that histamine-producing strains of this species have been isolated from any food). The restriction profiles of their genomes were analysed by PFGE, and seven lineages identified. The presence of the histidine decarboxylase gene (hdcA) was confirmed by PCR. The nucleotide sequence and genetic organisation of the histamine biosynthesis gene cluster (HDC) and its flanking regions are described for a representative strain (L. vaginalis IPLA11050).

  19. Isolation and characteristics of a novel biphenyl-degrading bacterial strain, Dyella ginsengisoli LA-4

    Institute of Scientific and Technical Information of China (English)

    LI Ang; QU Yuanyuan; ZHOU Jiti; GOU Min

    2009-01-01

    A novel biphenyl-degrading bacterial strain LA-4 was isolated from activated sludge. It was identified as Dyella ginsengisoli according to phylogenetic similarity of 16S rRNA gene sequence. This isolate could utilize biphenyl as sole source of carbon and energy, which degraded over 95 mg/L biphenyl within 36 h. The major metabolites formed from biphenyl, such as 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA) and benzoic acid, were identified by LC-MS. The crude cell extract of strain LA-4 exhibited the activity of 2,3-dihydroxybiphenyl 1,2-dioxygenase (2,3-DHBD) and the kinetic parameters were Km= 26.48 μmol/L and Vmax= 8.12 μmol/mg protein. A conserved region of the biphenyl dioxygenase gene bphA1 of strain LA-4 was amplified by PCR and confirmed by DNA sequencing.

  20. 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.

  1. 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.

  2. Occurrence OF Pectobacterium carotovorum strains isolated from potato soft rot in Morocco.

    Science.gov (United States)

    Terta, M; El Karkouri, A; Ait M'hand, R; Achbani, E; Barakate, M; Amdan, M; Annajar, B; El Hassouni, M; Val, F; Bouteau, F; Ennaji, M M

    2010-09-11

    Pectobacterium carotovorum subsp. carotovorum, Pectobacterium astrosepticum and Pectobacterium chrysanthemi are the soft rot tuber of potatoes pathogens (Solanum tuberosum). The aim of this study was to determine the occurrence of these pathogens in Moroccan regions producing potatoes. Fifty three isolates of Pectobacterium were isolated on medium Crystal Violet Pectate. The comparison of their bacteriological characteristics with standard strains allowed us to conclude that all the isolates belonged to the Pectobacterium. With regard to phenotype characteristics, the variability that was found included 32 typical Pectobacetrium carotovorum subsp. carotovorum, 3 typical Pectobacterium atrosepticum, and 18 atypical Pectobacterium carotovorum subsp. carotovorum. Three strains of the atypical group; showed that the biochemical properties overlap among the Pectobacterium carotovorum and Pectobacterium chrysanthemi. These data were needed molecular characterization. However, the PCR amplification of total genomic DNA of 53 isolates with the two primers Y1/Y2 and P143/P145 yielded an amplified fragment of the expected size (434 bp) only with Y1/Y2, indicated that all the isolates collected and tested belonged to the Pectobacterium carotovorum species. On the basis the pathogenicity tests, these strains revealed that they were pectinolytic, and showed differences in aggressiveness against potato and leaves of tobacco.

  3. 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.

  4. 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.

  5. Pathogenic properties of a Corynebacterium diphtheriae strain isolated from a case of osteomyelitis.

    Science.gov (United States)

    Peixoto, Renata Stavracakis; Hacker, Elena; Antunes, Camila Azevedo; Weerasekera, Dulanthi; Dias, Alexandre Alves de Souza de Oliveira; Martins, Carlos Alberto; Hirata, Raphael; Santos, Kátia Regina Netto Dos; Burkovski, Andreas; Mattos-Guaraldi, Ana Luíza

    2016-11-01

    Corynebacterium diphtheriae is typically recognized as a colonizer of the upper respiratory tract (respiratory diphtheria) and the skin (cutaneous diphtheria). However, different strains of Corynebacteriumdiphtheriae can also cause invasive infections. In this study, the characterization of a non-toxigenic Corynebacteriumdiphtheriae strain (designated BR-INCA5015) isolated from osteomyelitis in the frontal bone of a patient with adenoid cystic carcinoma was performed. Pathogenic properties of the strain BR-INCA5015 were tested in a Caenorhabditis elegans survival assay showing strong colonization and killing by this strain. Survival rates of 3.8±2.7 %, 33.6±7.3 % and 0 % were observed for strains ATCC 27010T, ATCC 27012 and BR-INCA5015, respectively, at day 7. BR-INCA5015 was able to colonize epithelial cells, showing elevated capacity to adhere to and survive within HeLa cells compared to other Corynebacteriumdiphtheriae isolates. Intracellular survival in macrophages (THP-1 and RAW 264.7) was significantly higher compared to control strains ATCC 27010T (non-toxigenic) and ATCC 27012 (toxigenic). Furthermore, the ability of BR-INCA5015 to induce osteomyelitis was confirmed by in vivo assay using Swiss Webster mice.

  6. Physiological Properties and Salmonella Growth Inhibition of Probiotic Bacillus Strains Isolated from Environmental and Poultry Sources.

    Science.gov (United States)

    Menconi, Anita; Morgan, Marion J; Pumford, Neil R; Hargis, Billy M; Tellez, Guillermo

    2013-01-01

    The objective of the present study was to describe the physiological properties of seven potential probiotic strains of Bacillus spp. Isolates were characterized morphologically, biochemically, and by 16S rRNA sequence analyses for identification. Tolerance to acidic pH, high osmotic concentrations of NaCl, and bile salts were tested. Isolates were also evaluated for their ability to metabolize different carbohydrates sources. The antimicrobial sensitivity profiles were determined. Inhibition of gastrointestinal Salmonella colonization in an avian model was also evaluated. Five strains of Bacillus were tolerant to acidic conditions (pH 2.0) and all strains were tolerant to a high osmotic pressure (NaCl at 6.5%). Moreover, all strains were able to tolerate concentration of 0.037% bile salts after 24 h of incubation. Three strains were able to significantly reduce Salmonella Typhimurium levels in the crop and in the ceca of broiler-type chickens. Among the 12 antibiotics tested for antibiotic resistance, all strains were resistant to bacitracin and susceptible to gentamycin, neomycin, ormethoprim, triple sulfa, and spectinomycin. Bacterial spore formers have been shown to prevent gastrointestinal diseases in animals and humans. The results obtained in this study show important characteristics to be evaluated when selecting Bacillus spp. candidates to be used as probiotics.

  7. Physiological Properties and Salmonella Growth Inhibition of Probiotic Bacillus Strains Isolated from Environmental and Poultry Sources

    Directory of Open Access Journals (Sweden)

    Anita Menconi

    2013-01-01

    Full Text Available The objective of the present study was to describe the physiological properties of seven potential probiotic strains of Bacillus spp. Isolates were characterized morphologically, biochemically, and by 16S rRNA sequence analyses for identification. Tolerance to acidic pH, high osmotic concentrations of NaCl, and bile salts were tested. Isolates were also evaluated for their ability to metabolize different carbohydrates sources. The antimicrobial sensitivity profiles were determined. Inhibition of gastrointestinal Salmonella colonization in an avian model was also evaluated. Five strains of Bacillus were tolerant to acidic conditions (pH 2.0 and all strains were tolerant to a high osmotic pressure (NaCl at 6.5%. Moreover, all strains were able to tolerate concentration of 0.037% bile salts after 24 h of incubation. Three strains were able to significantly reduce Salmonella Typhimurium levels in the crop and in the ceca of broiler-type chickens. Among the 12 antibiotics tested for antibiotic resistance, all strains were resistant to bacitracin and susceptible to gentamycin, neomycin, ormethoprim, triple sulfa, and spectinomycin. Bacterial spore formers have been shown to prevent gastrointestinal diseases in animals and humans. The results obtained in this study show important characteristics to be evaluated when selecting Bacillus spp. candidates to be used as probiotics.

  8. Genetic characterization of Vibrio vulnificus strains isolated from oyster samples in Mexico.

    Science.gov (United States)

    Guerrero, Abraham; Gómez Gil Rodríguez, Bruno; Wong-Chang, Irma; Lizárraga-Partida, Marcial Leonardo

    2015-01-01

    Vibrio vulnificus strains were isolated from oysters that were collected at the main seafood market in Mexico City. Strains were characterized with regard to vvhA, vcg genotype, PFGE, multilocus sequence typing (MLST), and rtxA1. Analyses included a comparison with rtxA1 reference sequences. Environmental (vcgE) and clinical (vcgC) genotypes were isolated at nearly equal percentages. PFGE had high heterogeneity, but the strains clustered by vcgE or vcgC genotype. Select housekeeping genes for MLST and primers that were designed for rtxA1 domains divided the strains into two clusters according to the E or C genotype. Reference rtxA1 sequences and those from this study were also clustered according to genotype. These results confirm that this genetic dimorphism is not limited to vcg genotyping, as other studies have reported. Some environmental C genotype strains had high similarity to reference strains, which have been reported to be virulent, indicating a potential risk for oyster consumers in Mexico City.

  9. 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.

  10. Production of conjugated linoleic acids by Lactobacillusplantarum strains isolated from naturally fermented Chinese pickles

    Institute of Scientific and Technical Information of China (English)

    Pei LIU; Sheng-rong SHEN; Hui RUAN; Qian ZHOU; Liu-liu MA; Guo-qing HE

    2011-01-01

    Naturally fermented pickles harbour many lactic acid bacteria (LAB).Forty-three LAB strains with conjugated linoleic acid (CLA)-producing ability were isolated from three naturally fermented pickle brines.Of these isolates,Ip15 identified as Lactobacillus plantarum by API 50 CHL system and full-length 16S rDNA sequence analysis exhibited the highest CLA-producing ability (26.1% conversion) at 48 h in de Man Rogosa Sharpe (MRS) broth in the presence of 100 μg/ml of linoleic acid (LA).Compared to other strains,L.plantarum strain Ip15 showed the highest tolerance upon increased levels of LA in the medium,I.e.,up to 600 pg/ml.This strain converted about 25% of LA into CLA isomers [predominantly cis-9,trans-11 CLA (9-CLA) and trans-10,cis-12 CLA (10-CLA)],of which 75% was 9-CLA.Interestingly,though the conversion rate of LA into CLA by Ip15 remained stable between 100 to 600 μg/ml LA levels in the medium,it dropped sharply at 1000 μg/ml.Taken together,the Ip15 strain displayed relatively high LA tolerance with higher conversion rate,which implies that this strain is a valuable candidate for enhancing the CLA content in food-sources like pickles.

  11. 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.

  12. A new variant of rabbit hemorrhagic disease virus G2-like strain isolated in China.

    Science.gov (United States)

    Hu, Bo; Fan, Zhiyu; Wang, Fang; Song, Yanhua; Wei, Houjun; Liu, Xing; Qiu, Rulong; Xu, Weizhong; Yuan, Wanzhe; Xue, Jiabin

    2016-04-02

    To investigate the genetic variability and evolution of rabbit hemorrhagic disease virus (RHDV) strains in China, VP60 gene sequences of eight new isolates collected from farms with RHD occurrences in China between 2009 and 2014 were analyzed, and compared with the reference sequence of the vaccine strain WF/China/2007. We conducted a comprehensive analysis of the Chinese RHDV strains, including hemagglutination tests, western blot and immunosassays of capsid proteins, and phylogenetic analysis, and identified a new distinct antigenic variant. Specifically, strain HB/2014 collected in North China was identified as a non-hemagglutinating strain, and belongs to the original RHDV (G1-G5) group. The other seven isolates were classified in genogroup G6 (RHDVa), which was widely distributed across China before 2014, and was thought to replace the earlier groups. Antigenic characterization of the VP60 genes revealed a large degree of nucleotide sequence divergence between HB/2014 and the other Chinese strains. However, the current vaccine showed complete cross-protection against HB/2014 challenge in inoculated rabbits. Collectively, these data provide new tools and insight for further understanding the molecular evolution of RHDV in China.

  13. Polymorphism of viral dsRNA in Xanthophyllomyces dendrorhous strains isolated from different geographic areas

    Directory of Open Access Journals (Sweden)

    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

  14. 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.

  15. Production of endoglucanase by the native strains of Streptomyces isolates in submerged fermentation

    Science.gov (United States)

    Chellapandi, P.; Jani, Himanshu M.

    2008-01-01

    Cellulase is a complex enzyme system, commercially produced by filamentous fungi under solid-state and submerged cultivation. It has wide applicability in textile, food and beverage industry for effective saccharification process. In this study, cellulolytic enzyme activity, particularly endoglucanase of 26 Streptomyces strains isolated from garden soil was examined, including two isolates selected on the basis of potential cellulolytic activity on Bennett’s agar medium. To enhance the endoglucanase formation in broth culture, different conditions including carbon and nitrogen sources, and growth conditions were tested. The maximum endoglucanase activity (11.25-11.90 U/mL) was achieved within 72-88 h in fermentation medium containing Tween-80, followed by phosphate sources. Both cellulolytic Streptomyces isolates gave almost equal quantity of enzyme in all trials. However the effect of medium ingredients on endoglucanase induction diverged with strains in some extent. PMID:24031191

  16. Plutonium interaction with a bacterial strain isolated from the waste isolation pilot plant (WIPP) environment

    Energy Technology Data Exchange (ETDEWEB)

    Strietelmeier, B.A.; Kraus, S.M.; Leonard, P.A.; Triay, I.R. [Los Alamos National Lab., NM (United States)] [and others

    1996-12-31

    This work was conducted as part of a series of experiments to determine the association and interaction of various actinides with bacteria isolated from the WIPP site. The majority of bacteria that exist at the site are expected to be halophiles, or extreme halophiles, due to the high concentration of salt minerals at the location. Experiments were conducted to determine the toxicity of plutonium-n-239, neptunium-237 and americium-243 to several species of these halophiles and the results were reported elsewhere. As an extension of these experiments, we report an investigation of the type of association that occurs between {sup 239}Pu and the isolate WIPP-1A, isolated by staff at Brookhaven National Laboratory, when grown in a high-salt, defined medium. Using scanning electron microscopy (SEM) techniques, we demonstrate a surface association of the {sup 239}Pu with the bacterial cells.

  17. Multidrug resistant Escherichia coli strains isolated from urine sample, University of Gondar Hospital, Northwest Ethiopia

    Institute of Scientific and Technical Information of China (English)

    Setegn Eshetie; Fentahun Tarekegn; Gemechu Kumera; Feleke Mekonnen

    2016-01-01

    Objective: To assess multidrug resistant (MDR) Escherichia coli (E. coli) isolates from patients with urinary tract infection. Methods: From February to June 2014, a cross sectional study was conducted among urinary tract infection patients at the University of Gondar Hospital. Culture and disk diffusion method were used for E. coli isolation and to determine the antibiotic susceptibility patterns. Data were entered and analyzed using SPSS version 20. P Results: A total of 112 E. coli isolates were identified and the rate of isolation was higher among female participants (28.7%; P = 0.03). Of the isolates, 104 (92.9%) were MDR E. coli; and the isolates showed high resistance rates towards ampicillin (99%), cotrimoxazole (69%), chloramphenicol (58.7%), gentamycin (56.7%) and ceftazidime (55.8%). However, comparative isolates showed low resistance rates to ciprofloxacin (1%), cefepime (8.7%), and ceftriaxone (11.5%). Moreover, resistance rates of MDR E. coli isolates were significantly higher than non-MDR strains for ceftazidime (55.8% versus 12.5%; P = 0.015), and ampicillin (99% versus 87.5%; P = 0.018). Conclusions: High prevalence of MDR E. coli isolates was observed in this study. Regular monitoring of antibiotic resistance rates is necessarily required to improve and revise empirical antibiotic therapy protocols.

  18. Botulinum toxin: bioweapon & magic drug.

    Science.gov (United States)

    Dhaked, Ram Kumar; Singh, Manglesh Kumar; Singh, Padma; Gupta, Pallavi

    2010-11-01

    Botulinum neurotoxins, causative agents of botulism in humans, are produced by Clostridium botulinum, an anaerobic spore-former Gram positive bacillus. Botulinum neurotoxin poses a major bioweapon threat because of its extreme potency and lethality; its ease of production, transport, and misuse; and the need for prolonged intensive care among affected persons. A single gram of crystalline toxin, evenly dispersed and inhaled, can kill more than one million people. The basis of the phenomenal potency of botulinum toxin is enzymatic; the toxin is a zinc proteinase that cleaves neuronal vesicle associated proteins responsible for acetylcholine release into the neuromuscular junction. As a military or terrorist weapon, botulinum toxin could be disseminated via aerosol or by contamination of water or food supplies, causing widespread casualties. A fascinating aspect of botulinum toxin research in recent years has been development of the most potent toxin into a molecule of significant therapeutic utility . It is the first biological toxin which is licensed for treatment of human diseases. In the late 1980s, Canada approved use of the toxin to treat strabismus, in 2001 in the removal of facial wrinkles and in 2002, the FDA in the United States followed suit. The present review focuses on both warfare potential and medical uses of botulinum neurotoxin.

  19. Characterization of trh2 harbouring Vibrio parahaemolyticus strains isolated in Germany.

    Directory of Open Access Journals (Sweden)

    Silke Bechlars

    Full Text Available 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.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 hemolytic activity. TRH variants were synthesized in a prokaryotic cell-free system and their hemolytic activity was analyzed. TRH1 was active against sheep erythrocytes while TRH2 variants were not active at all.Our study reveals a high diversity among trh positive V. parahaemolyticus strains. The function of TRH2 hemolysins and the role of the pseudogene Ψtrh as pathogenicity factors are questionable. To assess the pathogenic potential of V. parahaemolyticus strains a differentiation of trh variants and the detection of T3SS2β components like vopC would improve the V. parahaemolyticus diagnostics and could lead to a refinement of the risk assessment in food

  20. Probiotic Activity of a Bacterial Strain Isolated from Ancient Permafrost Against Salmonella Infection in Mice.

    Science.gov (United States)

    Fursova, O; Potapov, V; Brouchkov, A; Pogorelko, G; Griva, G; Fursova, N; Ignatov, S

    2012-09-01

    Bacillus cereus strain F, collected from relict permafrost located in Siberia, was analyzed for probiotic activity in the mouse Salmonella enterica model. Viable bacterial cells were found in frozen soils taken at Mammoth Mountain in Yakutia from a depth below the level of seasonal thawing. Geological data indicated the absence of a thawing within millions of years of deposited soils, which helped to ensure the ancient origin of our sample. According to DNA analysis, bacterial cells collected from the relict permafrost appeared to be B. cereus strain F. The morphology of these bacteria was analyzed using atomic force microscopy. B. cereus strain F was assessed as a nonpathogenic bacterium by evaluation of its pathogenicity. A S. enterica model is described in mice after per oral inoculation and serves as a model for the human carrier state. Using this model, probiotic activity by the bacterial strain isolated from the ancient permafrost has been shown against Salmonella infection in mice.

  1. Variable flocculation profiles of yeast strains isolated from cachaça distilleries.

    Science.gov (United States)

    Alvarez, Florencia; Correa, Lygia Fátima da Mata; Araújo, Thalita Macedo; Mota, Bruno Eduardo Fernandes; da Conceição, Luís Eduardo F Ribeiro; Castro, Ieso de Miranda; Brandão, Rogelio Lopes

    2014-11-03

    In cachaça production, the use of yeast cells as starters with predictable flocculation behavior facilitates the cell recovery at the end of each fermentation cycle. Therefore, the aim of this work was to explain the behavior of cachaça yeast strains in fermentation vats containing sugarcane through the determination of biochemical and molecular parameters associated with flocculation phenotypes. By analyzing thirteen cachaça yeast strains isolated from different distilleries, our results demonstrated that neither classic biochemical measurements (e.g., percentage of flocculation, EDTA sensitivity, cell surface hydrophobicity, and sugar residues on the cell wall) nor modern molecular approaches, such as polymerase chain reaction (PCR) and real-time PCR (q-PCR), were sufficient to distinctly classify the cachaça yeast strains according to their flocculation behavior. It seems that flocculation is indeed a strain-specific phenomenon that is difficult to explain and/or categorize by the available methodologies.

  2. Establishing the change in antibiotic resistance of Enterococcus faecium strains isolated from Dutch broilers by logistic regression and survival analysis

    NARCIS (Netherlands)

    Stegeman, J.A.; Vernooij, J.C.M.; Khalifa, O.A.; Broek, van den J.; Mevius, D.J.

    2006-01-01

    In this study, we investigated the change in the resistance of Enterococcus faecium strains isolated from Dutch broilers against erythromycin and virginiamycin in 1998, 1999 and 2001 by logistic regression analysis and survival analysis. The E. faecium strains were isolated from caecal samples that

  3. Isolation of Klebsiella pneumoniae strains with altered susceptibility to carbapenems not carbapenemase mediated

    Directory of Open Access Journals (Sweden)

    Franca Cian

    2009-12-01

    Full Text Available The spread of enterobacteria producing extended-spectrum ß-lactamases (ESBLs is sharply increasing in Italy, while the detection of isolates resistant to carbapenems is still sporadic. Isolates of Klebsiella pneumoniae resistant to all cephalosporins, aminoglycosides and fluoroquinolones have been isolated in Trieste since 2008. Because of the altered profile of resistance to carbapenems, these strains were reported as ESBL-negative and possible carbapenemases producer by the expert system, leaving tigecycline as the only therapeutic choice.The purpose of this study is the characterization of the mechanisms involved in resistance to carbapenems in these strains and the evaluation of a reliable and simple test for phenotypic confirmation of ESBL and/or carbapenemase production. 25 isolates of MDR K. pneumoniae were collected between October 2008 and May 2009, mainly from urinary samples of elderly patients hospitalized in medicine wards. Identification and susceptibility testing were performed using the Vitek 2 system.The double-disc (DD test was used to check the production of ESBLs, while imipenem and imipenem-EDTA synergy test was used to detect the production of metallo-ßlactamase (MBL. Carbapenemase activity was tested by an hydrolysis assay and the production of MBLs was also investigated by PCR. The DD synergy test highlighted the possible production of ESBLs in 18 out of 22 strains, considered as negative by Vitek. All ESBLs producers tested positive for the blaCTX-M-15 allele. Only one isolate was resistant to carbapenems and resulted positive for production of MBL by the phenotypic test.The crude extract showed carbapenemase activity inhibited by EDTA; PCR test gave positive result for a blaVIM-type allele. PCR analysis performed on representative isolates, followed by sequencing, showed that coding sequence of ompk35 was not functional. Results of this study confirmed the emergence of ESBL-positive strains of K. pneumoniae that

  4. In vitro evaluation of adhesion and aggregation abilities of four potential probiotic strains isolated from guppy (Poecilia reticulata

    Directory of Open Access Journals (Sweden)

    Aparna Balakrishna

    2013-10-01

    Full Text Available The aim of this work was to study the probiotic-related characteristics of four strains of bacteria isolated from the normal flora of the guppy, Poecilia reticulata. In vitro results showed that the strains, namely, MBTU_PB1, MBTU_PB2, MBTU_PB3 and MBTU_PB4 had higher adhesion abilities than the tested indicator strains. However, an association between the cell-surface hydrophobicity and the ability to adhere to the intestinal mucus was not observed for these strains. Further, the selected strains were strongly autoaggregating (autoaggregation percentage ≥ 80 and also showed strain-specific coaggregation abilities with the tested indicator strains.

  5. Diversity and Ecophysiology of New Isolates of Extremely Acidophilic CS2-Converting Acidithiobacillus Strains

    Science.gov (United States)

    Smeulders, Marjan J.; Pol, Arjan; Zandvoort, Marcel H.; Jetten, Mike S. M.

    2013-01-01

    Biofiltration of industrial carbon disulfide (CS2)-contaminated waste air streams results in the acidification of biofilters and therefore reduced performance, high water use, and increased costs. To address these issues, we isolated 16 extremely acidophilic CS2-converting Acidithiobacillus thiooxidans strains that tolerated up to 6% (vol/vol) sulfuric acid. The ecophysiological properties of five selected strains (2Bp, Sts 4-3, S1p, G8, and BBW1) were compared. These five strains had pH optima between 1 (2Bp) and 2 (S1p). Their affinities for CS2 ranged between 80 (G8) and 130 (2Bp) μM. Strains S1p, G8, and BBW1 had more hydrophobic cell surfaces and produced less extracellular polymeric substance than did strains 2Bp and Sts 4-3. All five strains converted about 80% of the S added as CS2 to S0 when CS2 was supplied in excess. The rate of S0 consumption varied between 7 (Sts 4-3) and 63 (S1p) nmol O2 min−1 ml culture−1. Low S0 consumption rates correlated partly with low levels of cell attachment to externally produced S0 globules. During chemostat growth, the relative amount of CS2 hydrolase in the cell increased with decreasing growth rates. This resulted in more S0 accumulation during CS2 overloads at low growth rates. Intermittent interruptions of the CS2 supply affected all five strains. Strains S1p, G8, and BBW1 recovered from 24 h of starvation within 4 h, and strains 2Bp and Sts 4-3 recovered within 24 h after CS2 was resupplied. We recommend the use of mixtures of Acidithiobacillus strains in industrial biofilters. PMID:23995926

  6. Technological properties of indigenous wine yeast strains isolated from wine production regions of Turkey.

    Science.gov (United States)

    Bağder Elmacı, Simel; Özçelik, Filiz; Tokatlı, Mehmet; Çakır, İbrahim

    2014-05-01

    The purpose of this study was to evaluate the important technological and fermentative properties of wine yeast strains previously isolated from different wine producing regions of Turkey. The determination of the following important properties was made: growth at high temperatures; fermentative capability in the presence of high sugar concentration; fermentation rate; hydrogen sulfide production; killer activity; resistance to high ethanol and sulfur dioxide; foam production; and enzymatic profiles. Ten local wine yeast strains belonging to Saccharomyces, and one commercial active dry yeast as a reference strain were evaluated. Fermentation characteristics were evaluated in terms of kinetic parameters, including ethanol yield (YP/S), biomass yield (YX/S), theoretical ethanol yield (%), specific ethanol production rate (qp; g/gh), specific glucose uptake rate (qs; g/gh), and the substrate conversion (%). All tested strains were able to grow at 37 °C and to start fermentation at 30° Brix, and were resistant to high concentrations of sulfur dioxide. 60 % of the strains were weak H2S producers, while the others produced high levels. Foam production was high, and no strains had killer activity. Six of the tested strains had the ability to grow and ferment at concentrations of 14 % ethanol. Except for one strain, all fermented most of the media sugars at a high rate, producing 11.0-12.4 % (v/v) ethanol. Although all but one strain had suitable characteristics for wine production, they possessed poor activities of glycosidase, esterase and proteinase enzymes of oenological interest. Nine of the ten local yeast strains were selected for their good oenological properties and their suitability as a wine starter culture.

  7. [The incidence of extended spectrum beta-lactamases in Proteus mirabilis strains isolated in 2007-2009].

    Science.gov (United States)

    Kwiecińska-Piróg, Joanna; Bogiel, Tomasz; Gospodarek, Eugenia

    2010-01-01

    Proteus sp. rods are opportunistic human pathogens, isolated mainly from urinary tract infections. They are naturally susceptible to most antimicrobials. However, acquisition of genes for extended spectrum beta-lactamases on plasmids and irrational antimicrobial treatment increase amount of multidrug resistant strains and lead to their selection. The aim of this study was to evaluate production of ESBLs by double disc synergy test and antimicrobial susceptibility of P mirabilis strains by disc diffusion method. Strains included into the study were isolated from patients of dr A. Jurasz University Hospital No 1 in Bydgoszcz between 2007 and 2009. P mirabilis strains were isolated mainly from urine. In this study 10,4%, 18,7% and 14,4% ESBL(+) P mirabilis strains were isolated in 2007, 2008, 2009, respectively. Resistance to majority of the investigated antimicrobials was observed in ESBL(+) P mirabilis strains.

  8. Virulence Genes and Antimicrobial Resistance Profiles of Pasteurella multocida Strains Isolated from Rabbits in Brazil

    Directory of Open Access Journals (Sweden)

    Thais Sebastiana Porfida Ferreira

    2012-01-01

    Full Text Available Pasteurella multocida is responsible for a wide range of diseases in domestic animals. In rabbits, the agent is related to nasal discharge, pneumonia, otitis media, pyometra, orchitis, abscess, and septicemia. One hundred and forty rabbits with respiratory diseases from four rabbitries in São Paulo State, Brazil were evaluated for the detection of P. multocida in their nasal cavities. A total of twenty-nine animals were positive to P. multocida isolation, and 46 strains were selected and characterized by means of biochemical tests and PCR. P. multocida strains were tested for capsular type, virulence genes, and resistance profile. A total of 45.6% (21/46 of isolates belonged to capsular type A, and 54.34% (25/46 of the isolates were untypeable. None of the strains harboured toxA or pfhA genes. The frequency of the other twenty genes tested was variable, and the data generated was used to build a dendrogram, showing the relatedness of strains, which were clustered according to origin. Resistance revealed to be more common against sulfonamides and cotrimoxazole, followed by erythromycin, penicillin, and amoxicillin.

  9. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules

    Directory of Open Access Journals (Sweden)

    Pilar eMartínez-Hidalgo

    2015-09-01

    Full Text Available Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation.In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  10. Isolation and characterization of a new cellulosome-producing Clostridium thermocellum strain.

    Science.gov (United States)

    Tachaapaikoon, Chakrit; Kosugi, Akihiko; Pason, Patthra; Waeonukul, Rattiya; Ratanakhanokchai, Khanok; Kyu, Khin Lay; Arai, Takamitsu; Murata, Yoshinori; Mori, Yutaka

    2012-02-01

    The anaerobic thermophilic bacterium, Clostridium thermocellum, is a potent cellulolytic microorganism that produces large extracellular multienzyme complexes called cellulosomes. To isolate C. thermocellum organisms that possess effective cellulose-degrading ability, new thermophilic cellulolytic strains were screened from more than 800 samples obtained mainly from agriculture residues in Thailand using microcrystalline cellulose as a carbon source. A new strain, C. thermocellum S14, having high cellulose-degrading ability was isolated from bagasse paper sludge. Cellulosomes prepared from S14 demonstrated faster degradation of microcrystalline cellulose, and 3.4- and 5.6-fold greater Avicelase activity than those from C. thermocellum ATCC27405 and JW20 (ATCC31449), respectively. Scanning electron microscopic analysis showed that S14 had unique cell surface features with few protuberances in contrast to the type strains. In addition, the cellulosome of S14 was resistant to inhibition by cellobiose that is a major end product of cellulose hydrolysis. Saccharification tests conducted using rice straw soaked with sodium hydroxide indicated the cellulosome of S14 released approximately 1.5-fold more total sugars compared to that of ATCC27405. This newly isolated S14 strain has the potential as an enzyme resource for effective lignocellulose degradation.

  11. Study of Klebsiella pneumoniae strains resistant to carbapenems isolated from blood in eastern Liguria

    Directory of Open Access Journals (Sweden)

    Giulia Carnesecchi

    2012-12-01

    Full Text Available Objectives. Study of multi-resistant Klebsiella pneumoniae strains isolated from blood cultures collected from in-patients of hospitals located in eastern Liguria, and evaluation of the susceptibility to carbapenems and other antibiotics by E-test and automated methods. Methods. At the Laboratory of Clinical Microbiology, of Lavagna Hospital in eastern Liguria, 397 Klebsiella pneumoniae strains were collected from in-patients from different wards of hospitals sites, during the year 2011. They included 115 isolates from blood cultures (aerobic and anaerobic and various biological materials. All strains were tested in the laboratory for their susceptibility to antibiotics. Results. Of the 115 strains of Klebsiella pneumoniae collected from blood cultures 59.1% showed resistance to imipenem, ertapenem, meropenem. Conclusions. The data show a high incidence of resistance to carbapenems in Klebsiella pneumoniae isolated from blood cultures.This is important to implement surveillance programs for control and prevention, but also reduce the intake of antibiotics when they are not strictly necessary.

  12. Induced systemic resistance against Botrytis cinerea by Micromonospora strains isolated from root nodules.

    Science.gov (United States)

    Martínez-Hidalgo, Pilar; García, Juan M; Pozo, María J

    2015-01-01

    Micromonospora is a Gram positive bacterium that can be isolated from nitrogen fixing nodules from healthy leguminous plants, where they could be beneficial to the plant. Their plant growth promoting activity in legume and non-legume plants has been previously demonstrated. The present study explores the ability of Micromonospora strains to control fungal pathogens and to stimulate plant immunity. Micromonospora strains isolated from surface sterilized nodules of alfalfa showed in vitro antifungal activity against several pathogenic fungi. Moreover, root inoculation of tomato plants with these Micromonospora strains effectively reduced leaf infection by the fungal pathogen Botrytis cinerea, despite spatial separation between both microorganisms. This induced systemic resistance, confirmed in different tomato cultivars, is long lasting. Gene expression analyses evidenced that Micromonospora stimulates the plant capacity to activate defense mechanisms upon pathogen attack. The defensive response of tomato plants inoculated with Micromonospora spp. differs from that of non-inoculated plants, showing a stronger induction of jasmonate-regulated defenses when the plant is challenged with a pathogen. The hypothesis of jasmonates playing a key role in this defense priming effect was confirmed using defense-impaired tomato mutants, since the JA-deficient line def1 was unable to display a long term induced resistance upon Micromonospora spp. inoculation. In conclusion, nodule isolated Micromonospora strains should be considered excellent candidates as biocontrol agents as they combine both direct antifungal activity against plant pathogens and the ability to prime plant immunity.

  13. Antimicrobial resistance in Salmonella strains clinically isolated in Hyogo, Japan (2009-2012).

    Science.gov (United States)

    Osawa, Kayo; Shigemura, Katsumi; Shimizu, Rika; Kato, Ayaka; Kimura, Mayuha; Katayama, Yuki; Okuya, Yuma; Yutaka, Shunichiro; Nishimoto, Akiko; Kishi, Akane; Fujiwara, Miki; Yoshida, Hiroyuki; Iijima, Yoshio; Fujisawa, Masato; Shirakawa, Toshiro

    2014-01-01

    The purpose of this study was to examine the in vitro susceptibilities to antimicrobial agents and genetic diversity of 195 clinical strains of Salmonella spp., which were isolated and examined for the extended-spectrum β-lactamase (ESBL) blaCTX-M gene and the presence of gyrA, gyrB, parC, and parE genes mutations in Hyogo, Japan, from 2009 to 2012. Forty-three of the 195 strains were antimicrobial resistant. Two Salmonella enterica subsp. enterica strains, 1 serovar Schwarzengrund, and 1 serovar Enteritidis were identified as ESBL-producing strains possessing blaCTX-M-15 and blaCTX-M-2, respectively. Among 8 nalidixic acid-resistant strains, 7 had mutations in gyrA alone or in gyrA and parC. In conclusion, we identified CTX-M ESBL-producing Salmonella clinical strains with multidrug resistance. Further studies are needed to monitor these serious drug-resistant Salmonella strains in Japan.

  14. Brewing characteristics of haploid strains isolated from sake yeast Kyokai No. 7.

    Science.gov (United States)

    Katou, Taku; Kitagaki, Hiroshi; Akao, Takeshi; Shimoi, Hitoshi

    2008-11-01

    Sake yeast exhibit various characteristics that make them more suitable for sake brewing compared to other yeast strains. Since sake yeast strains are Saccharomyces cerevisiae heterothallic diploid strains, it is likely that they have heterozygous alleles on homologous chromosomes (heterozygosity) due to spontaneous mutations. If this is the case, segregation of phenotypic traits in haploid strains after sporulation and concomitant meiosis of sake yeast strains would be expected to occur. To examine this hypothesis, we isolated 100 haploid strains from Kyokai No. 7 (K7), a typical sake yeast strain in Japan, and compared their brewing characteristics in small-scale sake-brewing tests. Analyses of the resultant sake samples showed a smooth and continuous distribution of analytical values for brewing characteristics, suggesting that K7 has multiple heterozygosities that affect brewing characteristics and that these heterozygous alleles do segregate after sporulation. Correlation and principal component analyses suggested that the analytical parameters could be classified into two groups, indicating fermentation ability and sake flavour.

  15. Isolation and characterization of aniline degradation slightly halophilic bacterium, Erwinia sp. Strain HSA 6.

    Science.gov (United States)

    Li, Junmin; Jin, Zexin; Yu, Binbin

    2010-07-20

    The isolated strain HSA6 is classified as Erwinia amylovora based on 16S rDNA sequence and the morphological and physiological properties. Strain HSA6 is the first reported E. amylovora in pure culture growing with aniline as sole electron donor and carbon source. The suitable pH for strain HSA6 is wide (from 5 to 11). Strain HSA6 is slightly halophilic with growth occurring at 0-10% (v/v) NaCl, and the suitable NaCl concentration for strain HSA6 is from 0% to 6%. The number of bacteria appeared to decrease with an increase in aniline concentration. The number of bacteria appeared to be constant as the wastewater concentration increased from 0% to 20%. However, the number of cells decreased with an increase in wastewater concentration from 30% to 50% and grew very slowly at 50%. The degradation rate of aniline was 100% at 0.5% aniline concentration after 24 h culture. The degradation rate of aniline was found to descend as the concentration of aniline increased from 0.5% to 3% and rose as the culture time increased. Strain HSA6 contains a plasmid with molecular weight higher than 42 kDA. Plasmid curing test and quantitative degradation test showed that strain requires the plasmid for aniline degradation. The gene cluster degrading aniline was determined in the plasmid by PCR amplification.

  16. Strain Diversity of Mycobacterium tuberculosis Isolates from Pulmonary Tuberculosis Patients in Afar Pastoral Region of Ethiopia

    Directory of Open Access Journals (Sweden)

    Mulugeta Belay

    2014-01-01

    Full Text Available Data on genotypic diversity of Mycobacterium tuberculosis complex (MTBC is important to understand its epidemiology, human adaptation, clinical phenotypes, and drug resistance. This study aimed to characterize MTBC clinical isolates circulating in a predominantly pastoralist area in Ethiopia, a country where tuberculosis is the second leading cause of mortality. Culture of sputum samples collected from a total of 325 pulmonary TB suspects was done to isolate MTBC. Spoligotyping was used to characterize 105 isolates from culture positive slopes and the result was compared with an international database. Forty-four spoligotype patterns were observed to correspond to 35 shared-types (SITs containing 96 isolates and 9 orphan patterns; 27 SITs containing 83 isolates matched a preexisting shared-type in the database, whereas 8 SITs (n=13 isolates were newly created. A total of 19 SITs containing 80 isolates were clustered within this study (overall clustering of 76.19%. Three dominant lineages (T, CAS, and Manu accounted for 76.19% of the isolates. SIT149/T3-ETH was one of the two most dominant sublineages. Unlike previous reports, we show that Manu lineage strains not only constitute a dominant lineage, but are also associated with HIV infection in Afar region of Ethiopia. The high level of clustering suggests the presence of recent transmission that should be further studied using additional genotyping markers.

  17. Genotyping of potentially pathogenic Acanthamoeba strains isolated from nasal swabs of healthy individuals in Peru.

    Science.gov (United States)

    Cabello-Vílchez, Alfonso Martín; Martín-Navarro, Carmen María; López-Arencibia, Atteneri; Reyes-Batlle, María; González, Ana C; Guerra, Humberto; Gotuzzo, Eduardo; Valladares, Basilio; Piñero, José E; Lorenzo-Morales, Jacob

    2014-02-01

    Free Living Amoebae (FLA) of Acanthamoeba genus are widely distributed in the environment and can be found in the air, soil and water; and have also been isolated from air-conditioning units. In humans, they are causative agents of a sight-threating infection of the cornea, Acanthamoeba keratitis (AK) and a fatal infection of the central nervous system known as Granulomatous Amoebic Encephalitis (GAE). In this study, a survey was conducted in order to determine the presence and pathogenic potential of free-living amoebae of Acanthamoeba genus in nasal swabs from individuals in two regions of Peru. Identification of isolates was based on cyst morphology and PCR-sequencing of the Diagnostic Fragment 3 to identify strains at the genotype level. The pathogenic potential of the isolates was also assayed using temperature and osmotolerance assays and extracellular proteases zymograms. The obtained results revealed that all isolated strains exhibited pathogenic potential. After sequencing the highly variable DF3 (Diagnostic Fragment 3) region in the 18S rRNA gene as previously described, genotype T4 was found to be the most common one in the samples included in this study but also genotype T15 was identified. To the best of our knowledge, this is the first study on the characterization of Acanthamoeba strains at the genotype level and the first report of genotype T4 and T15 in Peru.

  18. The isolation of multiple strains of Mycoplasma ovipneumoniae from individual pneumonic sheep lungs.

    Science.gov (United States)

    Ionas, G; Clarke, J K; Marshall, R B

    1991-11-01

    The heterogeneity of Mycoplasma ovipneumoniae isolates from the lungs of sheep with chronic non-progressive pneumonia (CNP) from the same flock raised the possibility that multiple isolates derived from one lung were not all identical. To test this hypothesis, thirty isolates were obtained from each of six pneumonic sheep lungs at slaughter. Four lungs had relatively severe lesions and from each of these, three or four strains of M. ovipneumonia, distinguishable by REA and in most cases by SDS-PAGE, were detected. From the lungs of each of two sheep with mild lesions, two strains of M. ovipneumoniae were detected. Four isolates from one lung were further examined by restriction endonuclease analysis (REA) using many restriction endonucleases. Those which differed with EcoRI also differed when other restriction endonucleases were used. However, partial digests occurred mainly with those restriction endonucleases which recognise cytosine-rich sequences. The presence of multiple strains of one species of microorganism in individual lesions is an unusual concept which may not be limited to one disease or to one host.

  19. Molecular screening of pathogenic Escherichia coli strains isolated from dairy neonatal calves in Cordoba province, Argentina.

    Science.gov (United States)

    Picco, Natalia Y; Alustiza, Fabrisio E; Bellingeri, Romina V; Grosso, María C; Motta, Carlos E; Larriestra, Alejandro J; Vissio, Claudina; Tiranti, Karina I; Terzolo, Horacio R; Moreira, Ana R; Vivas, Adriana B

    2015-01-01

    The aim of this study was to perform a current molecular characterization of bovine pathogenic Escherichia coli strains isolated from random samplings in Argentinean dairy farms. Rectal swabs were obtained from 395 (63.7%) healthy and 225 (36.3%) diarrheic calves, belonging to 45 dairy farms in Cordoba Province, Argentina. E. coli isolates were examined for virulence genes (f5, f41, f17, sta, stb, lt, eae, vt) using PCR and the prevalence of E. coli virulence profiles was spatially described in terms of spatial distribution. A total of 30.1% isolates were found to be positive for at least one of the virulence genes. Depending on the different gene combinations present, 11 virulence profiles were found. Most of the isolates analyzed had a single gene, and no combination of fimbrial and enterotoxin gene was predominant. There was no association between the frequency and distribution of E. coli virulence genes and calf health status. Most of the virulence profiles were compatible with ETEC strains and showed a homogeneous distribution over the sampled area. A clustering pattern for E. coli virulence profiles could not be recognized. This work provides updated information on the molecular characterization of pathogenic E. coli strains from dairy herds in Cordoba, Argentina. These findings would be important to formulate prevention programs and effective therapies for diarrhea in calves caused by E. coli.

  20. Genetic and biochemical diversity of Gardnerella vaginalis strains isolated from women with bacterial vaginosis.

    Science.gov (United States)

    Pleckaityte, Milda; Janulaitiene, Migle; Lasickiene, Rita; Zvirbliene, Aurelija

    2012-06-01

    Gardnerella vaginalis is considered a substantial player in the progression of bacterial vaginosis (BV). We analysed 17 G. vaginalis strains isolated from the genital tract of women diagnosed with BV to establish a potential link between genotypes/biotypes and the expression of virulence factors, vaginolysin (VLY) and sialidase, which are assumed to play a substantial role in the pathogenesis of BV. Amplified ribosomal DNA restriction analysis revealed two G. vaginalis genotypes. Gardnerella vaginalis isolates of genotype 2 appeared more complex than genotype 1 and were subdivided into three subtypes. Biochemical typing allowed us to distinguish four different biotypes. A great diversity of the level of VLY production among the isolates of G. vaginalis may be related to a different cytotoxicity level of the strains. We did not find any correlation between VLY production level and G. vaginalis genotype/biotype. In contrast, a link between G. vaginalis genotype and sialidase production was established. Our findings on the diversity of VLY expression level in different clinical isolates and linking sialidase activity with the genotype of G. vaginalis could help to evaluate the pathogenic potential of different G. vaginalis strains.

  1. Major clonal lineages in impetigo Staphylococcus aureus strains isolated in Czech and Slovak maternity hospitals.

    Science.gov (United States)

    Růžičková, Vladislava; Pantůček, Roman; Petráš, Petr; Machová, Ivana; Kostýlková, Karla; Doškař, Jiří

    2012-11-01

    One hundred and twenty-seven exfoliative toxin-producing (ET-positive) strains of Staphylococcus aureus collected in 23 Czech and one Slovak maternity hospitals from 1998 to 2011 were genotypically characterized by multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) profiling, spa gene polymorphism analysis, and ETA-converting prophage carriage, which resulted in the identification of 21 genotypes grouped into 4 clonal complexes (CC). Ninety-one isolates carried the eta gene alone whilst 12 isolates harboured only the etb gene. Two new, to date not defined, spa types (t6644 and t6645) and 2 novel sequence types (ST2194 and ST2195) were identified in the set of strains under study. The predominant CC121 occurred in 13 Czech hospitals. CC15, CC9, and ST88 (CC88) exclusively included eta gene-positive strains while the strains belonging to ST121 harboured the eta and/or etb genes. This study highlights not only significant genomic diversity among impetigo strains and the distribution of major genotypes disseminated in the Czech and Slovak maternity hospitals, but also reveals their impact in epidermolytic infections.

  2. Biochemical behavior of Trypanosoma cruzi strains isolated from mice submitted to specific chemotherapy

    Directory of Open Access Journals (Sweden)

    Jesila Pinto M. Marretto

    1994-12-01

    Full Text Available To investigate the influence of chemotherapy on the biochemical beha vior of Trypanosoma cruzi strains, three groups of mice were infected with one of three strains of T. cruzi of different biological and isoenzymic patterns (Peruvian, 21 SF and Colombian strains. Each group was subdivided into subgroups: 1 - treated with nifurtimox; 2 - treated with benznidazole and 3 - untreated infected controls. At the end of treatment, that lasted for 90 days, xenodiagnosis, sub inoculation of blood into new born mice and haemoculture were performed as tests of cure. From the positive tests, 22 samples of T. cruzi were isolated from all subgroups. Electrophoretic analysis of the isoenzymes PGM, GP1, ALAT and AS AT failed to show any difference between parasite strains isolated from treated and untreated mice, which indicates that no detectable clonal selection or parasite genetic markers alterations concerning the isoenzymes analysed have been determined by treatment with drugs of recognized antiparasitic effect, suggesting stability of the phenotypic characteristics of the three biological types of T. cruzi strains.

  3. Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated From Soils in Rize, Turkey

    Directory of Open Access Journals (Sweden)

    Elif SEVİM

    2015-09-01

    Full Text Available Fifteen Bacillus strains which were isolated from soil samples were examined for resistance to 17 different antibiotics (ampicillin, methicillin, erythromycin, norfloxacin, cephalotine, gentamycin, ciprofloxacin, streptomycin, tobramycin, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, vancomycin, oxacilin, neomycin, kanamycin and, novabiocin and to 10 different heavy metals (copper, lead, cobalt, chrome, iron, mercury, zinc, nickel, manganese and, cadmium and for the presence of plasmid DNA. A total of eleven strains (67% were resistant to at least one antibiotic. The most common resistance was observed against methicillin and oxacillin. The most resistance strains were found as Bacillus sp. B3 and Bacillus sp. B11. High heavy metal resistance against copper, chromium, zinc, iron and nickel was detected, but mercury and cobalt resistance was not detected, except for 3 strains (B3, B11, and B12 which showed mercury resistance. It has been determined that seven Bacillus strains have plasmids. The isolated plasmids were transformed into the Bacillus subtilis W168 and it was shown that heavy metal and antibiotic resistance determinants were carried on these plasmids. These results showed that there was a correlation between plasmid content and resistance for both antibiotic and heavy metal resistance

  4. Bacillus flexus strain As-12, a new arsenic transformer bacterium isolated from contaminated water resources.

    Science.gov (United States)

    Jebeli, Mohammad Ahmadi; Maleki, Afshin; Amoozegar, Mohammad Ali; Kalantar, Enayatollah; Izanloo, Hassan; Gharibi, Fardin

    2017-02-01

    A total of 14 arsenic-resistant bacteria were isolated from an arsenic-contaminated travertine spring water in the central district of Qorveh county, Kurdistan Province, Iran. One of strains designated As-12 was selected for further investigation because of its ability to transform arsenic. The strain was identified by cultural, morphological and biochemical tests, and 16S rRNA gene sequencing. Finally, the growth characteristics of the isolate were investigated in a chemically defined medium which included varied ranges of environmental factors such as pH, temperature and salinity. Moreover, the resistance of this strain to some heavy metals was evaluated. The bacterium was a Gram-positive, endospore-forming with all other characteristics of the genus Bacillus. It revealed maximum similarity at the 16S rRNA gene level with Bacillus flexus. The optimum growth of the strain was observed at 38 °C, pH 9 and 2% salinity. This strain was resistant to heavy metals such as zinc, chromium, lead, nickel, copper, mercuric and cadmium at concentrations of 15 mM, 15.5 mM, 11.5 mM, 12 mM, 11 mM, 5.5 mM, and 1 mM, respectively. The isolated bacterium was able to reduce As (V) to As (III) (about 28%) and oxidize As (III) to As (V) (about 45%) after 48 h of incubation at 37 °C. In conclusion, Bacillus flexus strain As-12, was identified as an arsenic transformer, for the first time.

  5. Antimicrobial susceptibilities of Listeria monocytogenes human strains isolated from 1970 to 2008 in Brazil

    Directory of Open Access Journals (Sweden)

    Cristhiane Moura Falavina dos Reis

    2011-04-01

    Full Text Available INTRODUCTION: Listeria monocytogenes is the causative agent of listeriosis, a foodborne illness that affects mainly pregnant women, the elderly and immunocompromised patients. The primary treatment is a combination of ampicillin with an aminoglycoside, in addition to a second-choice drug represented by chloramphenicol, erythromycin, tetracycline and rifampicin. The aim of this study was to analyze the antimicrobial susceptibility profile of strains isolated from human sources in the last four decades. METHODS: Sixty-eight strains were selected from the culture collection of the Laboratory of Bacterial Zoonoses/LABZOO/FIOCRUZ isolated in different regions of Brazil from 1970 to 2008 and primarily isolated from cerebrospinal fluid and blood culture. Susceptibility tests to antimicrobials drugs were evaluated using the criteria established by Soussy using the Kirby-Bauer method and E-Test strips were used to determine the minimum inhibitory concentration (MIC. RESULTS: Among the strains tested, serovar L4b (60.3% was the most prevalent, followed by serovar 1/2a (20.6%, 1/2b (13.2% and the more uncommon serovars 1/2c, 3b and 4ab (5.9%. All strains were susceptible to ampicillin, cephalothin, erythromycin, gentamicin, teicoplanin and vancomycin. Only one strain (1.5% showed resistance to rifampin, and two (3% were resistant to trimethoprim-sulfamethoxazole. MICs with values up to 2μg/ml reinforce the need for microbiological surveillance. CONCLUSIONS: The study demonstrated low prevalence of strains resistant to the antimicrobial drugs indicated in the treatment of human listeriosis. Monitoring antimicrobial resistance profile is still very important to determine adequate treatment, especially in immunocompromised patients.

  6. Tyrosol degradation via the homogentisic acid pathway in a newly isolated Halomonas strain from olive processing effluents

    OpenAIRE

    Liebgott, Pierre-Pol; Labat, Marc; Amouric, Agnès; Tholozan, Jean-Luc; Lorquin, Jean

    2008-01-01

    To isolate a new Halomonas sp. strain capable of degrading tyrosol, a toxic compound present in olive mill wastewater, through the homogentisic acid (HGA) pathway. A moderately halophilic Gram-negative bacterium belonging to the Halomonas genus and designated strain TYRC17 was isolated from olive processing effluents. This strain was able to completely degrade tyrosol (2-(p-hydroxyphenyl)-ethanol), a toxic compound found in such effluent. Tyrosol degradation begins by an oxidation to 4-hydrox...

  7. Molecular gene profiling of Clostridium botulinum group III and its detection in naturally contaminated samples originating from various European countries.

    Science.gov (United States)

    Woudstra, Cedric; Le Maréchal, Caroline; Souillard, Rozenn; Bayon-Auboyer, Marie-Hélène; Anniballi, Fabrizio; Auricchio, Bruna; De Medici, Dario; Bano, Luca; Koene, Miriam; Sansonetti, Marie-Hélène; Desoutter, Denise; Hansbauer, Eva-Maria; Dorner, Martin B; Dorner, Brigitte G; Fach, Patrick

    2015-04-01

    We report the development of real-time PCR assays for genotyping Clostridium botulinum group III targeting the newly defined C. novyi sensu lato group; the nontoxic nonhemagglutinin (NTNH)-encoding gene ntnh; the botulinum neurotoxin (BoNT)-encoding genes bont/C, bont/C/D, bont/D, and bont/D/C; and the flagellin (fliC) gene. The genetic diversity of fliC among C. botulinum group III strains resulted in the definition of five major subgroups named fliC-I to fliC-V. Investigation of fliC subtypes in 560 samples, with various European origins, showed that fliC-I was predominant and found exclusively in samples contaminated by C. botulinum type C/D, fliC-II was rarely detected, no sample was recorded as fliC-III or fliC-V, and only C. botulinum type D/C samples tested positive for fliC-IV. The lack of genetic diversity of the flagellin gene of C. botulinum type C/D would support a clonal spread of type C/D strains in different geographical areas. fliC-I to fliC-III are genetically related (87% to 92% sequence identity), whereas fliC-IV from C. botulinum type D/C is more genetically distant from the other fliC types (with only 50% sequence identity). These findings suggest fliC-I to fliC-III have evolved in a common environment and support a different genetic evolution for fliC-IV. A combination of the C. novyi sensu lato, ntnh, bont, and fliC PCR assays developed in this study allowed better characterization of C. botulinum group III and showed the group to be less genetically diverse than C. botulinum groups I and II, supporting a slow genetic evolution of the strains belonging to C. botulinum group III.

  8. Molecular serotyping of rough Salmonella strains isolated from Danish pork production

    DEFF Research Database (Denmark)

    Löfström, Charlotta; Boel, Jeppe; Sisic, E.;

    2013-01-01

    for subtyping is necessary and serotyping is one of the most commonly used approaches for Salmonella. Traditionally, serotyping is done by slide agglutination using antisera aiming at two cell surface antigens: somatic (O) lipopolysaccharides (LPS) and flagellar (H). Some Salmonella strains have incomplete LPS...... structures, referred to as rough strains, or do not express H antigens, therefore serotyping by agglutination cannot be performed on these isolates. This results in data gaps when subtyping data is needed. To overcome this obstacle, serotypes can be determined on DNA level, i.e. by determining the presence...

  9. 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....... Moreover, strain M17PTZA496 has a genome of 2.15 Mbp, notably larger than that of the others, determined by lateral gene transfer (including phage-mediated incorporation) and duplication events. Important technological characters, such as growth kinetics, bacteriocin production, acidification kinetics...

  10. Experimental infections with rifampicin-resistant Clostridium perfringens strains in broiler chickens using isolator facilities

    DEFF Research Database (Denmark)

    Pedersen, Karl; Bjerrum, Lotte; Nauerby, Birgitte;

    2003-01-01

    Experimental infection studies were carried out on the ability of three Clostridium perfringens type A rifampicin-resistant strains to colonize the intestinal tract of broiler chickens kept in isolators from 1-day-old. Various doses of C. perfringens were given orally at 22 days, 9 days or at 1 day...... from those receiving high doses, but for no longer than 13 days. In chicks infected at 1-day-old there was transient colonization up to 15 days, and the most persistent colonization was in a group given a fresh broth culture of unwashed cells, including extracellular products. Test strains were rapidly...

  11. Complete Genome Sequence of Clostridium septicum Strain CSUR P1044, Isolated from the Human Gut Microbiota.

    Science.gov (United States)

    Benamar, Samia; Cassir, Nadim; Caputo, Aurélia; Cadoret, Frédéric; La Scola, Bernard

    2016-01-01

    Clostridium septicum is one of the first pathogenic anaerobes to be identified. Here, we announce the genome draft sequence of C. septicum strain CSUR P1044 isolated from the gut of a healthy adult. Its chromosome genome consists of 3.2 Mbp with a plasmid of 32 Kbp. C. septicum strain CSUR P1044 has a G+C content of 27.5%, and is composed of 3,125 protein-coding genes together with 103 RNA genes, including 22 rRNA genes.

  12. Bacillus rubiinfantis sp. nov. strain mt2T, a new bacterial species isolated from human gut

    Directory of Open Access Journals (Sweden)

    M. Tidjiani Alou

    2015-11-01

    Full Text Available Bacillus rubiinfantis sp. nov. strain mt2T is the type strain of B. rubiinfantis sp. nov., isolated from the fecal flora of a child with kwashiorkor in Niger. It is Gram-positive facultative anaerobic rod belonging to the Bacillaceae family. We describe the features of this organism alongside the complete genome sequence and annotation. The 4 311 083 bp long genome (one chromosome but no plasmid contains 4028 protein-coding gene and 121 RNA genes including nine rRNA genes.

  13. Enterococcus faecalis as multidrug resistance strains in clinical isolates in Imam Reza Hospital in Kermanshah, Iran.

    Science.gov (United States)

    Mohammadi, F; Ghafourian, S; Mohebi, R; Taherikalani, M; Pakzad, I; Valadbeigi, H; Hatami, V; Sadeghifard, N

    2015-01-01

    The current study aimed to investigate the prevalence of vancomycin-resistant Enterococcus in E. faecalis and E. faecium and antimicrobial susceptibility patterns, then dominant genes responsible for vancomycin resistance were determined. For this propose, 180 clinical isolates of Enterococcus were subjected for identification and antibiotic susceptibility assay. Then, the gene responsible vancomycin resistant strains were determined. The results demonstrated the E. faecalis as a dominant Enterococcus. Resistance to erythromycin was dominant and multidrug resistance strains observed in E. faecalis. vanA was responsible for vancomycin resistance. In conclusion, a high rate of resistance to antibiotics in Enterococcus is clearly problematic, and a novel strategy is needed to decrease resistance in Enterococcus.

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

    Science.gov (United States)

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

    2012-09-01

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

  15. Molecular characterization of Acanthamoeba strains isolated from domestic dogs in Tenerife, Canary Islands, Spain.

    Science.gov (United States)

    Valladares, María; Reyes-Batlle, María; Martín-Navarro, Carmen M; López-Arencibia, Atteneri; Dorta-Gorrín, Alexis; Wagner, Carolina; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lorenzo-Morales, Jacob

    2015-06-01

    The present study describes two cases of Acanthamoeba infections (keratitis and ascites/peritonitis) in small breed domestic dogs in Tenerife, Canary Islands, Spain. In both cases, amoebic trophozoites were observed under the inverted microscope and isolated from the infected tissues and/or fluids, without detecting the presence of other viral, fungal or bacterial pathogens. Amoebae were isolated using 2 % non-nutrient agar plates and axenified for further biochemical and molecular analyses. Osmotolerance and thermotolerance assays revealed that both isolates were able to grow up to 37 °C and 1 M of mannitol and were thus considered as potentially pathogenic. Moreover, the strains were classified as highly cytotoxic as they cause more than 75 % of toxicity when incubated with two eukaryotic cell lines. In order to classify the strains at the molecular level, the diagnostic fragment 3 (DF3) region of the 18S rDNA of Acanthamoeba was amplified and sequenced, revealing that both isolates belonged to genotype T4. In both cases, owners of the animals did not allow any further studies or follow-up and therefore the current status of these animals is unknown. Furthermore, the isolation of these pathogenic amoebae should raise awareness with the veterinary community locally and worldwide.

  16. Detection of toxigenic Bacillus cereus strains isolated from vegetables in Mexico City.

    Science.gov (United States)

    Flores-Urbán, Karen A; Natividad-Bonifacio, Iván; Vázquez-Quiñones, Carlos R; Vázquez-Salinas, Carlos; Quiñones-Ramírez, Elsa Irma

    2014-12-01

    Bacillus cereus can cause diarrhea and emetic syndromes after ingestion of food contaminated with it. This ability is due to the production of enterotoxins by this microorganism, these being the hemolysin BL complex, which is involved in the diarrheal syndrome, and cereulide, which is responsible for the emetic syndrome. The detection of genes associated with the production of these toxins can predict the virulence of strains isolated from contaminated food. In this paper, we analyzed 100 samples of vegetables, 25 of each kind (broccoli, coriander, carrot, and lettuce) obtained from different markets in Mexico City and its metropolitan area. B. cereus was isolated in 32, 44, 84, and 68% of the samples of broccoli, carrot, lettuce, and coriander, respectively. The hblA gene (encoding one of the three subunits of hemolysin BL) was amplified in 100% of the B. cereus isolates, and the ces gene (encoding the cereulide) could not be amplified from any of them. This is the first report of B. cereus isolation from the vegetables analyzed in this work and, also, the first report in Mexico of the isolation from vegetables of strains with potential virulence. The results should serve as evidence of the potential risk of consuming these foods without proper treatment.

  17. Genotypic characterization, invasion index and antimicrobial resistance pattern in Listeria monocytogenes strains isolated from clinical samples

    Institute of Scientific and Technical Information of China (English)

    Behrooz Sadeghi Kalani; Abazar Pournajaf; Mansour Sedighi; Abbas Bahador; Gholamreza Irajian; Firuzeh Valian

    2015-01-01

    Objective: To evaluate antimicrobial resistance, invasion index and genetic profile in Listeriamonocytogenes isolated from clinical samples. Methods: At all, 170 clinical samples were collected from patients with spontaneous abortions hospitalized in Shariati hospital in Tehran during June 2010 to August 2013. Invasion index was determined using HeLa cells. The multiple-locus variable-number tandem-repeats analysis (MLVA) was used for evaluation of genetic relatedness. Results: Out of 14 L. monocytogenes isolates, 4 (28.57%), 2 (14.28%), 0 (0%), 5 (35.71%) and 3 (21.42%) were isolated from placental tissue, urine, blood, vaginal and rectal swabs, respectively. High resistance to penicillin and multidrug resistant were found amongst isolates. The invasion index was in the range of 0.001-0.007. Seven different types were obtained by MLVA assay and type 2 and 3 with 4 strains were the most frequent type. Strains isolated from the vagina and the placenta of the same type were also more resistant to penicillin. Conclusions: Since MLVA is a high-throughput screening method that is fairly inexpensive, easy to accomplish, rapid, and trustworthy, it is well suited to interlaboratory comparisons during epidemiological investigations. Also further studies of larger samples from a variety of sources such as food and animal specimens recommended comparing by MLVA method.

  18. Occurrence and characterization of plum pox virus strain D isolates from European Russia and Crimea.

    Science.gov (United States)

    Chirkov, Sergei; Ivanov, Peter; Sheveleva, Anna; Kudryavtseva, Anna; Prikhodko, Yuri; Mitrofanova, Irina

    2016-02-01

    Numerous plum pox virus (PPV) strain D isolates have been found in geographically distant regions of European Russia and the Crimean peninsula on different stone fruit hosts. Phylogenetic analysis of their partial and complete genomes suggests multiple introductions of PPV-D into Russia. Distinct natural isolates from Prunus tomentosa were found to bear unique amino acid substitutions in the N-terminus of the coat protein (CP) that may contribute to the adaptation of PPV-D to this host. Serological analysis using the PPV-D-specific monoclonal antibody 4DG5 provided further evidence that mutations at positions 58 and 59 of the CP are crucial for antibody binding.

  19. Isolation and Characterization of Frankia sp. Strain FaC1 Genes Involved in Nitrogen Fixation

    OpenAIRE

    Ligon, James M.; James P. Nakas

    1987-01-01

    Genomic DNA was isolated from Frankia sp. strain FaC1, an Alnus root nodule endophyte, and used to construct a genomic library in the cosmid vector pHC79. The genomic library was screened by in situ colony hybridization to identify clones of Frankia nitrogenase (nif) genes based on DNA sequence homology to structural nitrogenase genes from Klebsiella pneumoniae. Several Frankia nif clones were isolated, and hybridization with individual structural nitrogenase gene fragments (nifH, nifD, and n...

  20. Preliminary evaluation of probiotic potential of Lactobacillus plantarum strains isolated from Italian food products.

    Science.gov (United States)

    Turchi, Barbara; Mancini, Simone; Fratini, Filippo; Pedonese, Francesca; Nuvoloni, Roberta; Bertelloni, Fabrizio; Ebani, Valentina Virginia; Cerri, Domenico

    2013-10-01

    The aim of this study was to investigate some probiotic properties of 42 wild Lactobacillus plantarum strains isolated from different Italian foods of animal origin. The strains were first screened for their antibiotic resistance profile (chloramphenicol, erythromycin, gentamicin, and tetracycline), subsequently they were tested for their in vitro resistance to lysozyme (100 mg L⁻¹), low pH (3.0, 2.5 and 2.0) and bile salts (0.3, 0.5 and 1.0 %). Moreover, agglutination property was studied (adhesion to Saccharomyces cerevisiae cells), as well as the presence of bsh and msa genes. The strains with the best characteristics were subjected to a further trial in order to evaluate their ability to survive to multiple stresses over time (lysozyme, low pH and bile salts) and the effect of these treatments on adhesion to yeast cells. All the strains were susceptible to chloramphenicol, erythromycin and gentamicin, while 6 strains were excluded from further evaluation because of their resistant phenotype against tetracycline. All the strains were able to grow in presence of lysozyme, as well as in MRS broth at pH 3.0. Only 4 strains showed a growth rate lower than 80 % when grown in MRS broth at pH 2.5, while a relevant growth rate decrease was observed after exposure to pH 2.0. Bile salts didn't affect the viability of the L. plantarum cells. Twenty-one strains out of 33 tested strains were able to adhere to S. cerevisiae cells. Presence of both bsh and msa genes was detected in 6 strains. The strains resistant to all the stresses, positive to agglutination with S. cerevisiae and showing bsh and msa genes were selected for further evaluation and subjected to different stress treatments over time. The assessment of growth rates showed that exposure to lysozyme significantly increased low pH resistance in L. plantarum. This increase ranged from 2.35 to 15.57 %. The consequential lysozyme and low pH exposures didn't affect the growth rate values after bile salts treatment

  1. Isolation and characterization of a new Methanosarcina mazei strain GFJ07 from a mountain forest pond

    Directory of Open Access Journals (Sweden)

    Yi, Y.

    2012-01-01

    Full Text Available Aims: A new methanogenic strain, named GFJ07, was isolated from a pond of mountain forests in Guangxi, China. Cells grown in liquid culture tended to form aggregates with pseudosarcina-like or irregular shape.Methodology and Results: The optimum temperature, pH and NaCl concentration were 35 ℃, 7.0 and 0.5%, respectively. The isolate used methanol, trimethylamine, acetate and H2-CO2 as substrates. Analysis of the 16S rDNA sequences revealed strain GFJ07 showed the highest sequence similarity of 99.9% to Methanosarcina mazei.Conclusion, significance and impact of study: The cells were Gram positive and nonmotile. Most of single cell grew as a sausage-like clinder about 0.5 μm in diameter and 1.0 μm in length.

  2. Draft genome sequences of five Pseudomonas syringae pv. actinidifoliorum strains isolated in France.

    Science.gov (United States)

    Cunty, Amandine; Cesbron, Sophie; Briand, Martial; Carrère, Sébastien; Poliakoff, Françoise; Jacques, Marie-Agnès; Manceau, Charles

    2016-01-01

    Pseudomonas syringae pv. actinidifoliorum causes necrotic spots on the leaves of Actinidia deliciosa and Actinidia chinensis. P. syringae pv. actinidifoliorum has been detected in New Zealand, Australia, France and Spain. Four lineages were previously identified within the P. syringae pv. actinidifoliorum species group. Here, we report the draft genome sequences of five strains of P. syringae pv. actinidifoliorum representative of lineages 1, 2 and 4, isolated in France. The whole genomes of strains isolated in New Zealand, representative of P. syringae pv. actinidifoliorum lineages 1 and 3, were previously sequenced. The availability of supplementary P. syringae pv. actinidifoliorum genome sequences will be useful for developing molecular tools for pathogen detection and for performing comparative genomic analyses to study the relationship between P. syringae pv. actinidifoliorum and other kiwifruit pathogens, such as P. syringae pv. actinidiae.

  3. Draft genome sequences of five Pseudomonas syringae pv. actinidifoliorum strains isolated in France

    Directory of Open Access Journals (Sweden)

    Amandine Cunty

    Full Text Available ABSTRACT Pseudomonas syringae pv. actinidifoliorum causes necrotic spots on the leaves of Actinidia deliciosa and Actinidia chinensis. P. syringae pv. actinidifoliorum has been detected in New Zealand, Australia, France and Spain. Four lineages were previously identified within the P. syringae pv. actinidifoliorum species group. Here, we report the draft genome sequences of five strains of P. syringae pv. actinidifoliorum representative of lineages 1, 2 and 4, isolated in France. The whole genomes of strains isolated in New Zealand, representative of P. syringae pv. actinidifoliorum lineages 1 and 3, were previously sequenced. The availability of supplementary P. syringae pv. actinidifoliorum genome sequences will be useful for developing molecular tools for pathogen detection and for performing comparative genomic analyses to study the relationship between P. syringae pv. actinidifoliorum and other kiwifruit pathogens, such as P. syringae pv. actinidiae.

  4. Isolation of a new Mexican strain of Bacillus subtilis with antifungal and antibacterial activities.

    Science.gov (United States)

    Basurto-Cadena, M G L; Vázquez-Arista, M; García-Jiménez, J; Salcedo-Hernández, R; Bideshi, D K; Barboza-Corona, J E

    2012-01-01

    Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21) demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

  5. Isolation of a New Mexican Strain of Bacillus subtilis with Antifungal and Antibacterial Activities

    Directory of Open Access Journals (Sweden)

    M. G. L. Basurto-Cadena

    2012-01-01

    Full Text Available Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21 demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

  6. Surveillance of antimicrobial resistance in Escherichia coli strains isolated from pigs at Spanish slaughterhouses.

    Science.gov (United States)

    Teshager, T; Herrero, I A; Porrero, M C; Garde, J; Moreno, M A; Domínguez, L

    2000-07-01

    Antimicrobial resistance can make the efficient treatment of bacterial infections in humans and animals more difficult. Antimicrobial use in food animals may be one of the factors contributing to resistance. The Spanish surveillance network VAV has established a baseline of antimicrobial resistance in Escherichia coli strains from healthy pigs. Minimum inhibitory concentration and patterns of resistance to antimicrobials used in animals and humans were determined for 205 faecal strains isolated in a sampling frame of four slaughterhouses in Spain from 220 pigs in 1998. Higher levels of resistance were seen against antimicrobial agents authorised for use in food animals especially tetracycline, sulphonamides, trimethoprim and amoxycillin. All isolates were susceptible to antimicrobials employed mainly in humans such as ceftazidime, cefotaxime, imipenem, aztreonam and amikacin.

  7. Isolation and phylogenic identification of soil haloalkaliphilic strains in the former Texcoco Lake.

    Science.gov (United States)

    Soto-Padilla, Marisela Y; Valenzuela-Encinas, César; Dendooven, Luc; Marsch, Rodolfo; Gortáres-Moroyoqui, Pablo; Estrada-Alvarado, María Isabel

    2014-01-01

    A wide diversity of organisms exists in soil. Well-adapted groups can be found in extreme environments. A great economic and metabolic potential for extremozymes produced by organisms living at extreme environments has been reported. Extreme characteristics such as high salt content and high pH level make the soil of the former Texcoco Lake a unique place which has not been exploited. Therefore, in this study, 66 strains from soil of the former Texcoco Lake were isolated and phylogenetically analyzed using universal oligonucleotide primers. Different genera such as Kocuria, Micrococcus, Nesterenkonia, Halomonas, Salinicoccus, Kurthia, Gracilibacillus, and Bacillus were found. However, only 22 from all isolated strains were identified at specie level.

  8. A polyphasic taxonomic approach in isolated strains of Cyanobacteria from thermal springs of Greece.

    Science.gov (United States)

    Bravakos, Panos; Kotoulas, Georgios; Skaraki, Katerina; Pantazidou, Adriani; Economou-Amilli, Athena

    2016-05-01

    Strains of Cyanobacteria isolated from mats of 9 thermal springs of Greece have been studied for their taxonomic evaluation. A polyphasic taxonomic approach was employed which included: morphological observations by light microscopy and scanning electron microscopy, maximum parsimony, maximum likelihood and Bayesian analysis of 16S rDNA sequences, secondary structural comparisons of 16S-23S rRNA Internal Transcribed Spacer sequences, and finally environmental data. The 17 cyanobacterial isolates formed a diverse group that contained filamentous, coccoid and heterocytous strains. These included representatives of the polyphyletic genera of Synechococcus and Phormidium, and the orders Oscillatoriales, Spirulinales, Chroococcales and Nostocales. After analysis, at least 6 new taxa at the genus level provide new evidence in the taxonomy of Cyanobacteria and highlight the abundant diversity of thermal spring environments with many potential endemic species or ecotypes.

  9. Isolation and Identification of an Endophytic Strain with Antibiosis Ability from Davidia involucrate Brail.

    Institute of Scientific and Technical Information of China (English)

    NI Guo-rong; PAN Xiao-hua; TU Guo-quan; KUANG Fu-yuan; WEI Sai-jin

    2011-01-01

    [Objective]The aim was to isolate and identify an endophytic bacteria strain with antimicrobial activity from Davidia involucrate Brail.[Method]Endophytic strain with antibiosis ability was isolated from D.involucrate Brail by using cylinder-plate method.Then,it was identified through physiological and biochemical tests,16S rDNA homology analysis as well as some gene-specific sequence analysis.[Result]B221 stain had antimicrobial activity against a variety of rice plant pathogens,and it was identified as Bacillus subtilis.[Conclusion]This study enriches the research on endophyte within D.involucrate Brail,application of Bacillus bio-control,and therefore has laid a good foundation for the development of fungus used in biological control of crop pathogens.

  10. Sequence analysis of the rDNA intergenic spacer of Metarhizium strains isolated in Brazil

    Directory of Open Access Journals (Sweden)

    Fabiana Y. Yanaka-Schäfer

    2008-01-01

    Full Text Available To assess the extent of genetic variability of rDNA intergenic spacer (IGS in Metarhizium sp., 34 strains (27 isolated in Brazil were sequenced and analyzed together with an additional 20 Metarhizium anisopliae var. anisopliae sequences retrieved from GenBank. Overall, the global nucleotide diversity for the region under study was of 0.090, while for the Brazilian isolates it was only 0.016. Phylogenetic analyses showed four well-supported groups (A, B, C, and D, one of which (D has not been previously identified. All but one of the Brazilian strains cluster in this novel D phylogroup, suggesting that the genetic variation found in Brazil is a subset of the worldwide M. anisopiliae var. anisopliae variation.

  11. Coaggregation between Rhodococcus and Acinetobacter strains isolated from the food industry.

    Science.gov (United States)

    Møretrø, Trond; Sharifzadeh, Shahab; Langsrud, Solveig; Heir, Even; Rickard, Alexander H

    2015-07-01

    In this study, coaggregation interactions between Rhodococcus and Acinetobacter strains isolated from food-processing surfaces were characterized. Rhodococcus sp. strain MF3727 formed intrageneric coaggregates with Rhodococcus sp. strain MF3803 and intergeneric coaggregates with 2 strains of Acinetobacter calcoaceticus (MF3293, MF3627). Stronger coaggregation between A. calcoaceticus MF3727 and Rhodococcus sp. MF3293 was observed after growth in batch culture at 30 °C than at 20 °C, after growth in tryptic soy broth than in liquid R2A medium, and between cells in exponential and early stationary phases than cells in late stationary phase. The coaggregation ability of Rhodococcus sp. MF3727 was maintained even after heat and Proteinase K treatment, suggesting its ability to coaggregate was protein independent whereas the coaggregation determinants of the other strains involved proteinaceous cell-surface-associated polymers. Coaggregation was stable at pH 5-9. The mechanisms of coaggregation among Acinetobacter and Rhodococcus strains bare similarity to those displayed by coaggregating bacteria of oral and freshwater origin, with respect to binding between proteinaceous and nonproteinaceous determinants and the effect of environmental factors on coaggregation. Coaggregation may contribute to biofilm formation on industrial food surfaces, protecting bacteria against cleaning and disinfection.

  12. Investigation of genes involved in nisin production in Enterococcus spp. strains isolated from raw goat milk.

    Science.gov (United States)

    Perin, Luana Martins; Todorov, Svetoslav Dimitrov; Nero, Luís Augusto

    2016-09-01

    Different strains of Lactococcus lactis are capable of producing the bacteriocin nisin. However, genetic transfer mechanisms allow the natural occurrence of genes involved in nisin production in members of other bacterial genera, such as Enterococcus spp. In a previous study, nisA was identified in eight enterococci capable of producing antimicrobial substances. The aim of this study was to verify the presence of genes involved in nisin production in Enterococcus spp. strains, as well as nisin expression. The nisA genes from eight Enterococcus spp. strains were sequenced and the translated amino acid sequences were compared to nisin amino-acid sequences previously described in databases. Although containing nisin structural and maturation related genes, the enterococci strains tested in the present study did not present the immunity related genes (nisFEG and nisI). The translated sequences of nisA showed some point mutations, identical to those presented by Lactococcus strains isolated from goat milk. All enterococci were inhibited by nisin, indicating the absence of immunity and thus that nisin cannot be expressed. This study demonstrated for the first time the natural occurrence of nisin structural genes in Enterococcus strains and highlights the importance of providing evidence of a link between the presence of bacteriocin genes and their expression.

  13. Diffusely adherent Escherichia coli strains isolated from children and adults constitute two different populations

    Directory of Open Access Journals (Sweden)

    Mansan-Almeida Rosane

    2013-02-01

    Full Text Available Abstract Background Diffusely adherent Escherichia coli (DAEC have been considered a diarrheagenic category of E. coli for which several potential virulence factors have been described in the last few years. Despite this, epidemiological studies involving DAEC have shown inconsistent results. In this work, two different collections of DAEC possessing Afa/Dr genes, from children and adults, were studied regarding characteristics potentially associated to virulence. Results DAEC strains were recovered in similar frequencies from diarrheic and asymptomatic children, and more frequently from adults with diarrhea (P Citrobacter freundii strain have shown an improved ability to form biofilms in relation to the monocultures. Control strains have shown a greater diversity of Afa/Dr adhesins and higher frequencies of cellulose, TTSS, biofilm formation and induction of IL-8 secretion than strains from cases of diarrhea in children. Conclusions DAEC strains possessing Afa/Dr genes isolated from children and adults represent two different bacterial populations. DAEC strains carrying genes associated to virulence can be found as part of the normal microbiota present in asymptomatic children.

  14. [Isolation and characterization of a halotolerant p-nitroaniline degrading strain S8].

    Science.gov (United States)

    Song, Cai-Xia; Deng, Xin-Ping; Li, Tian; Xiao, Wei

    2014-03-01

    A bacterial strain S8 was isolated from pesticide-contaminated sludge, which showed ability of utilizing p-nitroaniline as the sole source of carbon and nitrogen for growth. Based on the morphology, the physiological and biochemical characteristics, and the 16S rDNA sequence analysis, the strain was identified as Bacillus subtilis. Strain S8 showed high efficiency in p-nitroaniline degradation. 65.6% and 55.8% of p-nitroaniline (60 mg x L(-1) and 120 mg x L(-1)) were degraded by this strain within 72 hours under the optimal conditions of 31degrees C and pH 6.0. Besides, strain S8 degraded 49.5% p-nitroaniline in 7% NaCl and 27.4% p-nitroaniline in 10% NaCl (72 h), which showed that the strain S8 had a high salinity tolerance. When the LC-MS method was used for identification of the biodegradation products, six kinds of degradation products were found, two of which were identified as phenol and hydroquinone. To date, this is the first report on the degradation of p-nitroaniline by Bacillus subtilis. These results suggest that S8 could be a potential candidate for treating p-nitroaniline-contaminated saline wastewater.

  15. Degradation of polyester polyurethane by a newly isolated soil bacterium, Bacillus subtilis strain MZA-75.

    Science.gov (United States)

    Shah, Ziaullah; Krumholz, Lee; Aktas, Deniz Fulya; Hasan, Fariha; Khattak, Mutiullah; Shah, Aamer Ali

    2013-11-01

    A polyurethane (PU) degrading bacterial strain MZA-75 was isolated from soil through enrichment technique. The bacterium was identified through 16S rRNA gene sequencing, the phylogenetic analysis indicated the strain MZA-75 belonged to genus Bacillus having maximum similarity with Bacillus subtilis strain JBE0016. The degradation of PU films by strain MZA-75 in mineral salt medium (MSM) was analyzed by scanning electron microscopy (SEM), fourier transform infra-red spectroscopy (FT-IR) and gel permeation chromatography (GPC). SEM revealed the appearance of widespread cracks on the surface. FTIR spectrum showed decrease in ester functional group. Increase in polydispersity index was observed in GPC, which indicates chain scission as a result of microbial treatment. CO2 evolution and cell growth increased when PU was used as carbon source in MSM in Sturm test. Increase in both cell associated and extracellular esterases was observed in the presence of PU indicated by p-Nitrophenyl acetate (pNPA) hydrolysis assay. Analysis of cell free supernatant by gas chromatography-mass spectrometry (GC-MS) revealed that 1,4-butanediol and adipic acid monomers were produced. Bacillus subtilis strain MZA-75 can degrade the soft segment of polyester polyurethane, unfortunately no information about the fate of hard segment could be obtained. Growth of strain MZA-75 in the presence of these metabolites indicated mineralization of ester hydrolysis products into CO2 and H2O.

  16. Isolation of a natural solopathogenic strain of Sporisorium reilianum f.sp. zeae (Ustilaginaceae, Basidiomycetes).

    Science.gov (United States)

    Sabbagh, S K; Naudan, M; Roux, C

    2008-01-01

    Sporisorium reilianum f.sp. zeae (Kühn) Langdon and Fullerton (Basidiomycota, Ustilaginaceae) is the causal agent of head smut of maize and sorghum. The parasitism is initiated by the fusion of two compatible sporidia which give rise to the formation of dikaryotic pathogen hyphae. However, in Ustilaginaceae, some fuzzy diploid strains could also be formed. These strains are solopathogen as they can infect a host in absence of crossing with a compatible haploid sporidia. A solopathogenic strain of S. refilianum was obtained using an original protocol. Sporidia were isolated from germinated teliospores and spread on solid medium to identify stable fuzzy solopathogenic strain. Confocal observations of the solopathogenic strain (SRZS1) after nucleus staining with propidium iodide indicates that they are formed by rounded shape cells which are monokaryotic. A CAPS approach was used to analysis the matb gene of S. reilianum. The presence of two matb loci in SRZS1 showed that this monocaryotic strain is diploid. The pathogenicity of SRZS1 was investigated by maize infection. Our results confirmed that SRZS1 is infectious, induces some typical symptoms in maize but could not sporulate and form sori.

  17. [Antibiotic resistance and biofilm formation in Staphylococcus saprophyticus strains isolated from urine].

    Science.gov (United States)

    Cernohorská, L; Votava, M

    2010-04-01

    Eighty-seven Staphylococcus saprophyticus strains isolated from urine of 87 patients with cystitis were examined in 2005-2009. All strains were tested for resistance to vancomycin, nitrofurantoin, doxycycline, oxacillin, amoxicillin/clavulanate, cefoxitin and sulfamethoxazole/trimethoprim and for biofilm formation by a modified Christensen method. None of the tested strains of S. saprophyticus showed resistance to vancomycin, while 2 strains (2.3 %) were resistant to nitrofurantoin, 9 (10.3%) to doxycycline, 20 (23.0 %) to oxacillin, 6 (6.9%) to amoxicillin/clavulanate, 6 (6.9%) to cefoxitin and 1 (1.1%) to sulfamethoxazole/trimethoprim. S. saprophyticus was detected as the causative agent of cystitis in 0.4 % of 20,375 culture positive urine samples analyzed in our laboratory between 2005 and 2009. Most 67 (77.0%) S. saprophyticus strains were recovered from women, particularly from young women. Biofilm formation was detected in 16 (18.4 %) out of 87 S. saprophyticus strains.

  18. Probiotic potential of lactobacillus strains isolated from sorghum-based traditional fermented food.

    Science.gov (United States)

    Rao, K Poornachandra; Chennappa, G; Suraj, U; Nagaraja, H; Raj, A P Charith; Sreenivasa, M Y

    2015-06-01

    Sorghum-based traditional fermented food was screened for potential probiotic lactic acid bacteria. The isolates were identified by biochemical, physiological and genetic methods. Species identification was done by 16s rRNA sequence analysis. The functional probiotic potential of the two Lactobacillus species viz., Lactobacillus plantarum (Lact. plantarum) and Lactobacillus pentosus (Lact. pentosus) was assessed by different standard parameters. The strains were tolerant to pH 2 for 1 h and resistant to methicillin, kanamycin, vancomycin and norfloxacin. Two (Lact. plantarum COORG-3 and Lact. pentosus COORG-8) out of eight isolates recorded the cell surface hydrophobicity to be 59.12 and 64.06%, respectively. All the strains showed tolerance to artificial duodenum juice (pH 2) for 3 h, positive for bile salt hydrolase test and negative for haemolytic test. The neutralized cell-free supernatant of the strains Lact. pentosus COORG-4, Lact. plantarum COORG-1, Lact. plantarum COORG-7, Lact. pentosus COORG-8 and Lact. plantarum COORG-3 showed good antibiofilm activity. Lact. pentosus COORG-8 exhibited 74% activity against Pseudomonas aeruginosa-MTCC 7903 and Lact. plantarum COORG-7 showed 68% inhibition of biofilm against Klebsiella pneumonia MTCC 7407. Three (Lact. plantarum COORG-7, Lact. pentosus COORG-5 and Lact. pentosus COORG 8) out of eight isolates exhibited a good antimicrobial activity against Listeria monocytogenes and five isolates (Lact. pentosus COORG 2, Lact. plantarum COORG 1, Lact. plantarum COORG 4, Lact. pentosus COORG 3 and Lact. plantarum COORG 6) are active against Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella pneumonia, Enterococcus faecalis. The study also evaluated the cholesterol lowering property of the Lactobacillus strains using hen egg yolk as the cholesterol source. The cholesterol in hen egg yolk was assimilated by 74.12 and 68.26% by Lact. plantarum COORG 4 and Lact. pentosus COORG 7

  19. Draft Genome Sequence of Corynebacterium amycolatum Strain ICIS 53 Isolated from a Female Urogenital Tract.

    Science.gov (United States)

    Gladysheva, Irina V; Cherkasov, Sergey V; Khlopko, Yuriy A; Plotnikov, Andrey O; Gogoleva, Natalya E

    2016-11-10

    This report describes the draft genome sequence of Corynebacterium amycolatum strain ICIS 53, isolated from the reproductive tract of a healthy woman. The size of the genome was 2,460,257 bp (58.98% G+C content). Annotation revealed 2,173 coding sequences, including 2,076 proteins, 7 rRNA genes, and 53 tRNA genes.

  20. Identification and Antibiosis of a Novel Actinomycete Strain RAF-11 Isolated From Iraqi Soil.

    Directory of Open Access Journals (Sweden)

    Rabah Forar Laidi

    2013-04-01

    Full Text Available A total of 35 actinomycetes strains were isolated from and around Baghdad, Iraq, at a depth of 5-10 m, by serial dilution agar plating method. Nineteen out of them showed noticeable antimicrobial activities against at least, to one of the target pathogens. Five among the nineteen were active against both Gram positive and Gram negative bacteria, yeasts and moulds. The most active isolate, strain RAF-11, based on its largest zone of inhibition and strong antifungal activity, especially against Candida albicans and Aspergillus niger, the causative of candidiasis and aspergillosis respectively, was selected for identification. Morphological and chemical studies indicated that this isolate belongs to the genus Streptomyces. Analysis of the 16S rDNA sequence showed a high similarity, 98 %, with the most closely related species, Streptomyces labedae NBRC 15864T/AB184704, S. erythrogriseus LMG 19406T/AJ781328, S. griseoincarnatus LMG 19316T/AJ781321 and S. variabilis NBRC 12825T/AB184884, having the closest match. From the taxonomic features, strain RAF-11 matched with S. labedae, in the morphological, physiological and biochemical characters, however it showed significant differences in morphological characteristics with this nearest species, S. labedae, which encourage us to consider our starin as a novel isolate and was given the suggested name, Streptomyces labedae strain RAF-11. ISP-4 broth medium supplemented with glucose and soybean powder at concentrations of 1g % and 0.1g % as carbon and nitrogen sources respectively, for 120h incubation at 28 °C, increased the active compounds production, where we recorded a strong activity against yeasts, 42mm inhibition zone against Candida albicans, 41mm against C. pseudotropicalis, 40mm against C. tropicalis, followed by 38mm against Rhodotorula minota and Aspergillus niger then, 35mm against both Aspergillus flavus and Bacillus subtilis. N-butanol was best solvent for antibiotic extraction compared to

  1. Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticum strains isolated from poultry

    Directory of Open Access Journals (Sweden)

    Gerchman Irena

    2011-08-01

    Full Text Available Abstract The macrolide class of antibiotics, including tylosin and tilmicosin, is widely used in the veterinary field for prophylaxis and treatment of mycoplasmosis. In vitro susceptibility testing of 50 strains of M. gallisepticum isolated in Israel during the period 1997-2010 revealed that acquired resistance to tylosin as well as to tilmicosin was present in 50% of them. Moreover, 72% (13/18 of the strains isolated from clinical samples since 2006 showed acquired resistance to enrofloxacin, tylosin and tilmicosin. Molecular typing of the field isolates, performed by gene-target sequencing (GTS, detected 13 molecular types (I-XIII. Type II was the predominant type prior to 2006 whereas type X, first detected in 2008, is currently prevalent. All ten type X strains were resistant to both fluoroquinolones and macrolides, suggesting selective pressure leading to clonal dissemination of resistance. However, this was not a unique event since resistant strains with other GTS molecular types were also found. Concurrently, the molecular basis for macrolide resistance in M. gallisepticum was identified. Our results revealed a clear-cut correlation between single point mutations A2058G or A2059G in domain V of the gene encoding 23S rRNA (rrnA, MGA_01 and acquired macrolide resistance in M. gallisepticum. Indeed, all isolates with MIC ≥ 0.63 μg/mL to tylosin and with MIC ≥ 1.25 μg/mL to tilmicosin possess one of these mutations, suggesting an essential role in decreased susceptibility of M. gallisepticum to 16-membered macrolides.

  2. Draft Genome Sequence of a Chitinophaga Strain Isolated from a Lignocellulose Biomass-Degrading Consortium

    Science.gov (United States)

    Kishi, Luciano T.; Lopes, Erica M.; Fernandes, Camila C.; Fernandes, Gabriela C.; Sacco, Lais P.; Carareto Alves, Lucia M.

    2017-01-01

    ABSTRACT Chitinophaga comprises microorganisms capable of degrading plant-derived carbohydrates, serving as a source of new tools for the characterization and degradation of plant biomass. Here, we report the draft genome assembly of a Chitinophaga strain with 8.2 Mbp and 7,173 open reading frames (ORFs), isolated from a bacterial consortium that is able to degrade lignocellulose. PMID:28104646

  3. Genome Sequence of Marine Bacterium Idiomarina sp. Strain 28-8, Isolated from Korean Ark Shells.

    Science.gov (United States)

    Kim, Woo-Jin; Kim, Young-Ok; Kim, Dong-Gyun; Nam, Bo-Hye; Kong, Hee Jeong; Jung, Hyungtaek; Lee, Sang-Jun; Kim, Dong-Wook; Kim, Dae-Soo; Chae, Sung-Hwa

    2013-10-03

    Idiomarina sp. strain 28-8 is an aerobic, Gram-negative, flagellar bacterium isolated from the bodies of ark shells (Scapharca broughtonii) collected from underwater sediments in Gangjin Bay, South Korea. Here, we present the draft genome sequence of Idiomarina sp. 28-8 (2,971,606 bp, with a G+C content of 46.9%), containing 2,795 putative coding sequences.

  4. Antibiotic resistance of Staphylococcus aureus strains isolated from fish processing factory workers

    OpenAIRE

    1988-01-01

    One hundred and twenty two strains of Staphylococcus aureus isolated from throats and palms of 39 workers from 6 fish processing factories situated in and around Cochin were tested for their sensitivity to nine commonly used antibiotics-ampicillin, chloramphenicol, erythromycin, kanamycin, neomycin, penicillin, polymyxin-B, streptomycin and tetracycline. Highest percentage of resistance was observed towards ampicillin followed by penicillin i.e. 64.75% and 59.84%. Resistance towards other ant...

  5. Chlamydia pneumoniae clinical isolate from gingival crevicular fluid: a potential atherogenic strain

    Directory of Open Access Journals (Sweden)

    Simone eFilardo

    2015-11-01

    Full Text Available 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.

  6. Genome Sequence of Lactobacillus paracasei Strain LC-Ikematsu, Isolated from a Pineapple in Okinawa, Japan

    Science.gov (United States)

    Saitoh, Seikoh; Aoyama, Hiroaki; Shinzato, Naoya; Yamamoto, Norikuni; Arita, Masanori; Ikematsu, Shinya

    2017-01-01

    ABSTRACT The draft genome sequence of Lactobacillus paracasei strain LC-Ikematsu, isolated from a pineapple in Okinawa, was determined. The total length of the 87 contigs was 3.08 Mb with a G+C content of 46.2% and 2,946 coding sequences. The genome analysis revealed its biosynthetic ability of 11 amino acids. PMID:28153910

  7. Characterization of Vibrio cholerae Strains Isolated from the Nigerian Cholera Outbreak in 2010.

    Science.gov (United States)

    Dupke, Susann; Akinsinde, Kehinde A; Grunow, Roland; Iwalokun, Bamidele A; Olukoya, Daniel K; Oluwadun, Afolabi; Velavan, Thirumalaisamy P; Jacob, Daniela

    2016-10-01

    We examined clinical samples from Nigerian patients with acute watery diarrhea for Vibrio cholerae during the 2010 cholera outbreak. A total of 109 suspected isolates were characterized, but only 57 V. cholerae strains could be confirmed using multiplex real-time PCR as well as rpoB sequencing and typed as V. cholerae O:1 Ogawa biotype El Tor. This finding highlighted the need for accurate diagnosis of cholera in epidemic countries to implement life-saving interventions.

  8. Comparative genotyping of Clostridium thermocellum strains isolated from biogas plants: genetic markers and characterization of cellulolytic potential.

    Science.gov (United States)

    Koeck, Daniela E; Zverlov, Vladimir V; Liebl, Wolfgang; Schwarz, Wolfgang H

    2014-07-01

    Clostridium thermocellum is among the most prevalent of known anaerobic cellulolytic bacteria. In this study, genetic and phenotypic variations among C. thermocellum strains isolated from different biogas plants were determined and different genotyping methods were evaluated on these isolates. At least two C. thermocellum strains were isolated independently from each of nine different biogas plants via enrichment on cellulose. Various DNA-based genotyping methods such as ribotyping, RAPD (Random Amplified Polymorphic DNA) and VNTR (Variable Number of Tandem Repeats) were applied to these isolates. One novel approach - the amplification of unknown target sequences between copies of a previously discovered Random Inserted Mobile Element (RIME) - was also tested. The genotyping method with the highest discriminatory power was found to be the amplification of the sequences between the insertion elements, where isolates from each biogas plant yielded a different band pattern. Cellulolytic potentials, optimal growth conditions and substrate spectra of all isolates were characterized to help identify phenotypic variations. Irrespective of the genotyping method used, the isolates from each individual biogas plant always exhibited identical patterns. This is suggestive of a single C. thermocellum strain exhibiting dominance in each biogas plant. The genotypic groups reflect the results of the physiological characterization of the isolates like substrate diversity and cellulase activity. Conversely, strains isolated across a range of biogas plants differed in their genotyping results and physiological properties. Both strains isolated from one biogas plant had the best specific cellulose-degrading properties and might therefore achieve superior substrate utilization yields in biogas fermenters.

  9. The hydrolytic enzymes produced by fungi strains isolated from the sand and soil of recreational areas

    Science.gov (United States)

    Kurnatowski, Piotr; Wójcik, Anna; Błaszkowska, Joanna; Góralska, Katarzyna

    2016-10-01

    The pathogenicity of fungi depends on, inter alia, the secretion of hydrolytic enzymes. The aim of this study was to determine the enzymatic activity of yeasts and yeast-like fungi isolated from children’s recreation areas, and compare the results with literature data of strains obtained from patients with mycoses. The enzymatic activity of 96 strains was assessed using an API ZYM kit (bioMerieux, France) and their biotypes were established. The fungal species were found to produce from 16 to 19 hydrolases: the most active were: leucine arylamidase (e5), acid phosphatase (e10), alkaline phosphatase (e1), naphthol-AS-BI-phosphohydrolase (e11), esterase – C4 (e2), β-galac - tosidase (e13) and β-glucosidase (e16). In addition, 13 biotypes characteristic of particular species of fungi were defined. Most strains could be categorized as biotypes C2 – 39.5% and A – 26%. The examined fungal strains isolated from recreational areas have selected biochemical characteristics i.e. production of hydrolases, which demonstrate their pathogenicity. They produce a number of enzymes which are also present in strains isolated from patients with mycoses, including: leucine arylamidase (e5), acid phosphatase (e10), naphthol-AS-BI-phosphohydrolase (e11) and alkaline phosphatase (e1). The biotypes identified in the course of this study (A, B3, B4, C1, C6 and D3) have been also reported in cases of fungal infection. Therefore, the fungi present in the sand and soil of recreational have pathogenic properties and are possible factors of fungal infection among children.

  10. Indigenous strains of Lactobacillus isolated from the Istrian cheese as potential starter cultures

    Directory of Open Access Journals (Sweden)

    Nataša Hulak

    2016-11-01

    Full Text Available Istrian ewe’s milk cheese is an autochthonous product that is manufactured for generations on small family farms in the Croatian peninsula Istria. Traditional Istrian cheese is made from unpasteurized ewe’s milk, without the addition of starter cultures. Consequently, the specific flavour and texture of the Istrian cheese is owed to metabolic processes of indigenous microflora of which Lactobacillus species play pivotal role. Characterisation and selection of indigenous lactobacilli may result in the potential use of selected strains as starter, bioprotective or even probiotic cultures. This study focuses on potential use of Lactobacillus plantarum and Lactobacillus casei isolated from traditional Istrian cheese as starter cultures, by using methods that determine their proteolytic, lipolytic, antimicrobial and haemolytic potential, as well as their ability of acidification, autoaggregation and survival in simulated gastrointestinal conditions. Our results indicated that from 12 representative strains most revealed a low or moderate proteolytic activity as well as absence of lipolytic and haemolytic activities. From 12 strains, 5 of them showed a medium to strong acidification ability and lowered the pH of milk below 5.00 after 24 hours of incubation. Furthermore, almost all isolates exhibited antimicrobial activity against Serratia marcescens, and lowest number of isolates showed antimicrobial activity against Staphylococcus aureus and Listeria innocua. The studied Lactobacillus strains revealed high survival rate in a simulated oral cavity and duodenum conditions, while the survival ability in a simulated gastric conditions was much lower. Ability to aggregate was low for all tested strains, after 3 hours and after 5 hours of incubation.

  11. Characteristics and antimicrobial activity of Bacillus subtilis strains isolated from soil.

    Science.gov (United States)

    Todorova, Sevdalina; Kozhuharova, Lubka

    2010-07-01

    Antagonistic Bacillus strains were isolated from soil and analyzed for the purpose of determining whether they could be used as natural biological agents. Primary in vitro screening for antagonism of the isolates was performed against five phytopathogenic mould fungi. Strains TS 01 and ZR 02 exhibited the most pronounced inhibitory effects. They were identified as Bacillus subtilis on the basis of their morphological, cultural and physiology-biochemical properties as well as their hierarchical cluster analysis conducted by means of computer program SPSS. The antimicrobial activity of the strains from cultural medium and sterile filtrate were determined in vitro against a great number of predominantly phytopathogenic fungi and bacteria. TS 01 and ZR 02 strains exhibited very broad and at the same time degree varying antibiotic spectra of activities against both Gram-positive and Gram-negative microorganisms. Many of them were tested against sensitivity to the antimicrobial action of B. subtilis for the very first time. B. subtilis TS 01 and ZR 02 showed highest antifungal activity (sterile zone in diameter over 37 mm) against Alternaria solani, Botrytis cinerea, Monilia linhartiana 869, Phytophthora cryptogea 759/1 and Rhizoctonia sp. The most sensitive bacterial species were found to be Pseudomonas syringae pv. tomato Ro and Xanthomonas campestris with sterile zones 48.0 and 50.0 mm in diameter, respectively. The latter draws a conclusion that the isolated and identified Bacillus subtilis strains are promising natural biocontrol agents and should be further studied and tested for control of numerous plant diseases.

  12. Detection of β-Lactamases and Outer Membrane Porins among Klebsiella pneumoniae Strains Isolated in Iran

    Directory of Open Access Journals (Sweden)

    Ali Hashemi

    2014-01-01

    Full Text Available This descriptive study was accomplished on 83 K. pneumoniae strains isolated from two hospitals in Tehran, Iran. Antibiotic susceptibility tests were performed by disc diffusion and broth microdilution methods. ESBLs, MBL, Amp-C, and KPC producing strains were detected by phenotypic confirmatory test, combination disk diffusion test (CDDT, Amp-C detection kit, and modified Hodge test, respectively. OXA-48, NDM-1, and CTX-M-15 genes were detected by PCR and sequencing methods. The outer membrane porins such as OmpK35 and OmpK36 were analysed by SDS-PAGE, PCR, and sequencing methods. From 83 K. pneumoniae isolates, 48 (57.5%, 3 (3.5%, 23 (28%, and 5 (6% were ESBL, MBL, Amp-C, and KPC positive, respectively. The CTX-M-15 gene was detected in 30 (62.5% and OXA-48 gene was found in 2 (4.1% of the 48 ESBL-producing isolates. Two isolates harboured both OXA-48 and CTX-M-15; NDM-1 gene was not detected in this study. Outer membrane porin, OmpK35, was detected in 30 (62.5% of 48 ESBL-producing isolates while OmpK36 was found in 35 (72.91% of 48 ESBL-producing isolates. In this study, fosfomycin and tigecycline were more effective than other antibiotics. The high prevalence of β-lactamase-producing K. pneumoniae detected in this study is of great concern, which requires infection control measures including antibacterial management and identification of β-lactamases-producing isolates.

  13. rpoB Mutations in Multidrug-Resistant Strains of Mycobacterium tuberculosis Isolated in Italy

    Science.gov (United States)

    Pozzi, G.; Meloni, M.; Iona, E.; Orrù, G.; Thoresen, O. F.; Ricci, M. L.; Oggioni, M. R.; Fattorini, L.; Orefici, G.

    1999-01-01

    Mutations of rpoB associated with rifampin resistance were studied in 37 multidrug-resistant (MDR) clinical strains of Mycobacterium tuberculosis isolated in Italy. At least one mutated codon was found in each MDR strain. It was always a single-base substitution leading to an amino acid change. Nine different rpoB alleles, three of which had not been reported before, were found. The relative frequencies of specific mutations in this sample were different from those previously reported from different geographical areas, since 22 strains (59.5%) carried the mutated codon TTG in position 531 (Ser→Leu) and 11 (29.7%) had GAC in position 526 (His→Asp). PMID:10074552

  14. Characterization and phylogenetic analysis of a phenanthrene-degrading strain isolated from oil-contaminated soil

    Institute of Scientific and Technical Information of China (English)

    XIA Ying; MIN Hang; LU Zhen-mei; YE Yang-fang

    2004-01-01

    Bacterium strain EVA17 was isolated from an oil-contaminated soil, and identified as Sphingononas sp.based on analysis of 16S rDNA sequence, cellular fatty acid composition and physiological-chemical tests. The salicylate hydroxylase and catechol 2, 3-dioxygenase (C23O) were detected in cell-free lysates, suggesting a pathway for phenanthrene catabolism via salicylate and catechol. Alignment showed that both of the C23O and GST genes of the strain EVA17 had high similarity with homologues of strains from genus Sphingomonas. The phylogenetic analysis based on 16S rDNA and C23O gene sequence indicated that EVA17 should be classified into genus Sphingomonas, although the two phylogenetic trees were slightly different from each other. The results of coamplification and sequence determination indicated that GST gene should be located upstream of the C23O gene.

  15. Isolation and characterization of a novel strain of Natrinema containing a bop gene

    Institute of Scientific and Technical Information of China (English)

    XU Xue-wei; WU Min; HUANG Wei-da

    2005-01-01

    A novel member of extremely halophilic archaea, strain AJ2, was isolated from Ayakekum Lake located in Altun Mountain National Nature Reserve of Xinjiang Uygur Autonomous Region in China. The strain A J2 requires at least 10% (w/v)NaCl and grows 10% to 30% (optimum at 20%). Phylogenetic analysis based on 16S rDNA sequence comparison revealed that strain A J2 clustered to three Natrinema species with less than 97.7% sequence similarities, suggesting A J2 is a novel member of Natrinema. A bacteriorhodopsin-encoding (bop) gene was subsequently detected in the A J2 genome using the polymerase chain reaction technique. The cloning and sequencing of a 401 base pairs fragment indicated the deduced amino acid sequence of bop from A J2 is different from that reported for bacteriorhodopsins. This is the first reported detection of a bop gene in Natrinema.

  16. Isolation and characterization of a bacterial strain that efficiently degrades sex steroid hormones

    Institute of Scientific and Technical Information of China (English)

    JI Shulan; LIU Zhipei; LIU Zhipeng; REN Haiyan

    2007-01-01

    A bacterial strain,ZY3,growing on sex steroid hormones as the sole source of carbon and energy was isolated from the sewage treatment plant of a prophylactic steroids factory.ZY3 degrades the 3-methoxy-17β-hyclroxy-1,3,5(10),8(9)-δ-4-estren (MHE).This strain was preliminarily identified as Raoultella sp.ZY3 according to its morphology and its 16S rRNA gene sequence.During the experimental period (72 h),the optimum temperature,pH and 3-MHE concentration for the degradation of hydride by the strain ZY3 were 35℃,10 and 10 mg/L,respectively.The degradation rate of the sex steroid hormones increased to 87% and 85% after the addition of maltose and peptone,respectively.

  17. Environmental carbonate chemistry selects for phenotype of recently isolated strains of Emiliania huxleyi

    Science.gov (United States)

    Rickaby, Rosalind E. M.; Hermoso, Michaël; Lee, Renee B. Y.; Rae, Benjamin D.; Heureux, Ana M. C.; Balestreri, Cecilia; Chakravarti, Leela; Schroeder, Declan C.; Brownlee, Colin

    2016-05-01

    Coccolithophorid algae, particularly Emiliania huxleyi, are prolific biomineralisers that, under many conditions, dominate communities of marine eukaryotic plankton. Their ability to photosynthesise and form calcified scales (coccoliths) has placed them in a unique position in the global carbon cycle. Contrasting reports have been made with regards to the response of E. huxleyi to ocean acidification. Therefore, there is a pressing need to further determine the fate of this key organism in a rising CO2 world. In this paper, we investigate the phenotype of newly isolated, genetically diverse, strains of E. huxleyi from UK Ocean Acidification Research Programme (UKOA) cruises around the British Isles, the Arctic, and the Southern Ocean. We find a continuum of diversity amongst the physiological and photosynthetic parameters of different strains of E. huxleyi morphotype A under uniform, ambient conditions imposed in the laboratory. This physiology is best explained by adaptation to carbonate chemistry in the former habitat rather than being prescribed by genetic fingerprints such as the coccolithophore morphology motif (CMM). To a first order, the photosynthetic capacity of each strain is a function of both aqueous CO2 availability, and calcification rate, suggestive of a link between carbon concentrating ability and calcification. The calcification rate of each strain is related linearly to the natural environmental [CO32-] at the site of isolation, but a few exceptional strains display low calcification rates at the highest [CO32-] when calcification is limited by low CO2 availability and/or a lack of a carbon concentrating mechanism. We present O2-electrode measurements alongside coccolith oxygen isotopic composition and the uronic acid content (UAC) of the coccolith associated polysaccharide (CAP), that act as indirect tools to show the differing carbon concentrating ability of the strains. The environmental selection revealed amongst our recently isolated strain

  18. Isolation and characterization of a novel native Bacillus thuringiensis strain BRC-HZM2 capable of degrading chlorpyrifos.

    Science.gov (United States)

    Wu, Songqing; Peng, Yan; Huang, Zhangmin; Huang, Zhipeng; Xu, Lei; Ivan, Gelbič; Guan, Xiong; Zhang, Lingling; Zou, Shuangquan

    2015-03-01

    Studies were carried out to isolate chlorpyrifos degrading Bacillus thuringiensis (Bt) strains from chlorpyrifos-contaminated samples. Six Bt strains (isolation rate 2.7%) were isolated by modified sodium acetate antibiotic heat treatment, and one novel strain (BRC-HZM2) was selected for further analysis. Phenotype and phylogeny analysis of this strain was conducted on the basis of biochemical reactions, antibiotic sensitivity, 16s rRNA genes, plasmid profile, insecticidal crystal protein profiles, and PCR-RFLP for cry and cyt genes. The degradation rate of chlorpyrifos in liquid culture was estimated during 48 h of incubation for the isolate BRC-HZM2. More than 50% of the initial chlorpyrifos concentration degraded within 12 h, 88.9% after 48 h. These results highlight the potential of the Bt strain for biological control and the bioremediation of environments contaminated with chlorpyrifos.

  19. Bioremediation potential of a highly mercury resistant bacterial strain Sphingobium SA2 isolated from contaminated soil.

    Science.gov (United States)

    Mahbub, Khandaker Rayhan; Krishnan, Kannan; Megharaj, Mallavarapu; Naidu, Ravi

    2016-02-01

    A mercury resistant bacterial strain, SA2, was isolated from soil contaminated with mercury. The 16S rRNA gene sequence of this isolate showed 99% sequence similarity to the genera Sphingobium and Sphingomonas of α-proteobacteria group. However, the isolate formed a distinct phyletic line with the genus Sphingobium suggesting the strain belongs to Sphingobium sp. Toxicity studies indicated resistance to high levels of mercury with estimated EC50 values 4.5 mg L(-1) and 44.15 mg L(-1) and MIC values 5.1 mg L(-1) and 48.48 mg L(-1) in minimal and rich media, respectively. The strain SA2 was able to volatilize mercury by producing mercuric reductase enzyme which makes it potential candidate for remediating mercury. ICP-QQQ-MS analysis of Hg supplemented culture solutions confirmed that almost 79% mercury in the culture suspension was volatilized in 6 h. A very small amount of mercury was observed to accumulate in cell pellets which was also evident according to ESEM-EDX analysis. The mercuric reductase gene merA was amplified and sequenced. The deduced amino acid sequence demonstrated sequence homology with α-proteobacteria and Ascomycota group.

  20. Bioremediation potential of a newly isolate solvent tolerant strain Bacillus thermophilus PS11

    Directory of Open Access Journals (Sweden)

    PAYEL SARKAR

    2012-01-01

    Full Text Available The increased generation of solvent waste has been stated as one of the most critical environmental problems. Though microbial bioremediation has been widely used for waste treatment but their application in solvent waste treatment is limited since the solvents have toxic effects on the microbial cells. A solvent tolerant strain of Bacillus thermophilus PS11 was isolated from soil by cyclohexane enrichment. Transmission electron micrograph of PS11 showed convoluted cell membrane and accumulation of solvents in the cytoplasm, indicating the adaptation of the bacterial strain to the solvent after 48h of incubation. The strain was also capable of growing in presence of wide range of other hydrophobic solvents with log P-values below 3.5. The isolate could uptake 50 ng/ml of uranium in its initial 12h of growth, exhibiting both solvent tolerance and metal resistance property. This combination of solvent tolerance and metal resistance will make the isolated Bacillus thermophilus PS11 a potential tool for metal bioremediation in solvent rich wastewaters.

  1. Comparative Genomic Analysis of Lactococcus garvieae Strains Isolated from Different Sources Reveals Candidate Virulence Genes

    Directory of Open Access Journals (Sweden)

    Eiji Miyauchi

    2012-01-01

    Full Text Available Lactococcus garvieae is a major pathogen for fish. Two complete (ATCC 49156 and Lg2 and three draft (UNIUD074, 8831, and 21881 genome sequences of L. garvieae have recently been released. We here present the results of a comparative genomic analysis of these fish and human isolates of L. garvieae. The pangenome comprised 1,542 core and 1,378 dispensable genes. The sequenced L. garvieae strains shared most of the possible virulence genes, but the capsule gene cluster was found only in fish-pathogenic strain Lg2. The absence of the capsule gene cluster in other nonpathogenic strains isolated from mastitis and vegetable was also confirmed by PCR. The fish and human isolates of L. garvieae contained the specific two and four adhesin genes, respectively, indicating that these adhesion proteins may be involved in the host specificity differences of L. garvieae. The discoveries revealed by the pangenomic analysis may provide significant insights into the biology of L. garvieae.

  2. Complete genome sequence of a novel Plum pox virus strain W isolate determined by 454 pyrosequencing.

    Science.gov (United States)

    Sheveleva, Anna; Kudryavtseva, Anna; Speranskaya, Anna; Belenikin, Maxim; Melnikova, Natalia; Chirkov, Sergei

    2013-10-01

    The near-complete (99.7 %) genome sequence of a novel Russian Plum pox virus (PPV) isolate Pk, belonging to the strain Winona (W), has been determined by 454 pyrosequencing with the exception of the thirty-one 5'-terminal nucleotides. This region was amplified using 5'RACE kit and sequenced by the Sanger method. Genomic RNA released from immunocaptured PPV particles was employed for generation of cDNA library using TransPlex Whole transcriptome amplification kit (WTA2, Sigma-Aldrich). The entire Pk genome has identity level of 92.8-94.5 % when compared to the complete nucleotide sequences of other PPV-W isolates (W3174, LV-141pl, LV-145bt, and UKR 44189), confirming a high degree of variability within the PPV-W strain. The isolates Pk and LV-141pl are most closely related. The Pk has been found in a wild plum (Prunus domestica) in a new region of Russia indicating widespread dissemination of the PPV-W strain in the European part of the former USSR.

  3. Phosphate solubilizing ability of Emericella nidulans strain V1 isolated from vermicompost.

    Science.gov (United States)

    Bhattacharya, Satya Sunder; Barman, Soma; Ghosh, Ranjan; Duary, Raj Kumar; Goswami, Linee; Mandal, Narayan C

    2013-10-01

    Phosphorus is one of the key factors that regulate soil fertility. Its deficiencies in soil are largely replenished by chemical fertilizers. The present study was aimed to isolate efficient phosphate solubilizing fungal strains from Eisenia fetida vermicompost. Out of total 30 fungal strains the most efficient phosphate solubilizing one was Emericella (Aspergillus) nidulans V1 (MTCC 11044), identified by custom sequencing of beta-tubulin gene and BLAST analysis. This strain solubilized 13 to 36% phosphate from four different rock phosphates. After three days of incubation of isolated culture with black Mussorie phosphate rock, the highest percentage of phosphate solubilization was 35.5 +/- 1.01 with a pH drop of 4.2 +/- 0.09. Kinetics of solubilization and acid production showed a linear relationship until day five of incubation. Interestingly, from zero to tenth day of incubation, solubility of soil phosphate increased gradually from 4.31 +/- 1.57 to 13.65 +/- 1.82 (mg kg(-1)) recording a maximum of 21.23 +/- 0.54 on day 45 in respect of the V1 isolate. Further, enhanced phosphorus uptake by Phaseolus plants with significant pod yield due to soil inoculation of Emericella nidulans V1 (MTCC 11044), demonstrated its prospect as an effective biofertilizer for plant growth.

  4. Effect of Antimicrobials on Salmonella Spp. Strains Isolated from Poultry Processing Plants

    Directory of Open Access Journals (Sweden)

    L Mion

    Full Text Available ABSTRACT The routine use of antimicrobials in animal production for the treatment of infections, disease prevention, or as growth promoters is a predisposing factor for the development and dissemination of antimicrobial resistance. In food industries, sanitizers are used for the control of microbial colonization, and their efficacy depends on contact time and on the dilution of the products used. The present study assessed the effect of 12 antimicrobials and four commercial sanitizers on 18 Salmonella spp. strains isolated from poultry processing plants. None of the evaluated antimicrobials was 100% effective against the tested Salmonella spp. strains; however, 94% of the isolates were susceptible to ciprofloxacin, 77% to amoxicillin + clavulanic acid and to ampicillin, and 72% to enrofloxacin, whereas 100% of the isolates were resistant to penicillin G, 16% to tetracycline, and 11% to sulfonamide. The tested Salmonella spp. strains were 100% inhibited by peracetic acid after five minutes of contact, 0.5% by quaternary ammonium after 15 minutes, and 85.7% by chlorhexidine after 15 minutes. The results indicate the importance of testing of efficacy of antimicrobials used in animal production and in public health to monitor their action and the development of resistance.

  5. Genome sequencing and annotation of Laceyella sacchari strain GS 1-1, isolated from hot spring, Chumathang, Leh, India

    Directory of Open Access Journals (Sweden)

    Navjot Kaur

    2014-12-01

    Full Text Available We report the 3.3-Mb draft genome of Laceyella sacchari strain GS 1-1, isolated from hot spring water sample, Chumathang, Leh, India. Draft genome of strain GS 1-1 consists of 3, 324, 316 bp with a G + C content of 48.8% and 3429 predicted protein coding genes and 75 RNAs. Geobacillus thermodenitrificans strain NG80-2, Geobacillus kaustophilus strain HTA426 and Geobacillus sp. Strain G11MC16 are the closest neighbors of the strain GS 1-1.

  6. Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity

    Directory of Open Access Journals (Sweden)

    Feng Pan

    2016-06-01

    Full Text Available Abstract The aim of this paper is to identify and investigate an endophytic fungus (strain 28 that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass. Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB as the base culture medium, fermentation for 4–8 d (initial pH: 7.5, followed by extraction with EtOAc. The extract was stable at temperatures up to 80 °C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata.

  7. Identification and adhesion profile of Lactobacillus spp. strains isolated from poultry.

    Science.gov (United States)

    Rocha, Ticiana Silva; Baptista, Ana Angelita Sampaio; Donato, Tais Cremasco; Milbradt, Elisane Lenita; Okamoto, Adriano Sakai; Andreatti Filho, Raphael Lucio

    2014-01-01

    In the aviculture industry, the use of Lactobacillus spp. as a probiotic has been shown to be frequent and satisfactory, both in improving bird production indexes and in protecting intestine against colonization by pathogenic bacteria. Adhesion is an important characteristic in selecting Lactobacillus probiotic strains since it impedes its immediate elimination to enable its beneficial action in the host. This study aimed to isolate, identify and characterize the in vitro and in vivo adhesion of Lactobacillus strains isolated from birds. The Lactobacillus spp. was identified by PCR and sequencing and the strains and its adhesion evaluated in vitro via BMM cell matrix and in vivo by inoculation in one-day-old birds. Duodenum, jejunum, ileum and cecum were collected one, four, 12 and 24 h after inoculation. The findings demonstrate greater adhesion of strains in the cecum and an important correlation between in vitro and in vivo results. It was concluded that BMM utilization represents an important technique for triage of Lactobacillus for subsequent in vivo evaluation, which was shown to be efficient in identifying bacterial adhesion to the enteric tract.

  8. Antimicrobial resistance and virulence gene profiles in P. multocida strains isolated from cats.

    Science.gov (United States)

    Ferreira, Thais Sebastiana Porfida; Felizardo, Maria Roberta; de Gobbi, Debora Dirani Sena; Moreno, Marina; Moreno, Andrea Micke

    2015-03-01

    Cats are often described as carriers of Pasteurella multocida in their oral microbiota. This agent is thought to cause pneumonia, conjunctivitis, rhinitis, gingivostomatitis, abscess and osteonecrosis in cats. Human infection with P. multocida has been described in several cases affecting cat owners or after cat bites. In Brazil, the cat population is approximately 21 million animals and is increasing, but there are no studies of the presence of P. multocida in the feline population or of human cases of infection associated with cats. In this study, one hundred and ninety-one healthy cats from owners and shelters in São Paulo State, Brazil, were evaluated for the presence of P. multocida in their oral cavities. Twenty animals were positive for P. multocida , and forty-one strains were selected and characterized by means of biochemical tests and PCR. The P. multocida strains were tested for capsular type, virulence genes and resistance profile. A total of 75.6% (31/41) of isolates belonged to capsular type A, and 24.4% (10/41) of the isolates were untypeable. None of the strains harboured toxA, tbpA or pfhA genes. The frequencies of the other genes tested were variable, and the data generated were used to build a dendrogram showing the relatedness of strains, which were clustered according to origin. The most common resistance profile observed was against sulfizoxazole and trimethoprim-sulphamethoxazole.

  9. Antimicrobial resistance and virulence gene profiles in P. multocida strains isolated from cats

    Directory of Open Access Journals (Sweden)

    Thais Sebastiana Porfida Ferreira

    2015-03-01

    Full Text Available Cats are often described as carriers of Pasteurella multocida in their oral microbiota. This agent is thought to cause pneumonia, conjunctivitis, rhinitis, gingivostomatitis, abscess and osteonecrosis in cats. Human infection with P. multocida has been described in several cases affecting cat owners or after cat bites. In Brazil, the cat population is approximately 21 million animals and is increasing, but there are no studies of the presence of P. multocida in the feline population or of human cases of infection associated with cats. In this study, one hundred and ninety-one healthy cats from owners and shelters in São Paulo State, Brazil, were evaluated for the presence of P. multocida in their oral cavities. Twenty animals were positive for P. multocida, and forty-one strains were selected and characterized by means of biochemical tests and PCR. The P. multocida strains were tested for capsular type, virulence genes and resistance profile. A total of 75.6% (31/41 of isolates belonged to capsular type A, and 24.4% (10/41 of the isolates were untypeable. None of the strains harboured toxA, tbpA or pfhA genes. The frequencies of the other genes tested were variable, and the data generated were used to build a dendrogram showing the relatedness of strains, which were clustered according to origin. The most common resistance profile observed was against sulfizoxazole and trimethoprim-sulphamethoxazole.

  10. [Characterization of S gene of a strain of hantavirus isolated from Apodemus peninsulae in Heilongjiang Province].

    Science.gov (United States)

    Chen, Lu-Fei; Chen, Shu-Hong; Wang, Kai-Li; Zhang, Jing; Li, Ji-Hong

    2012-09-01

    In order to study the molecular characterization of the hantavirus isolated from Apodemus peninsulae in Heilongjiang Province, the S gene of a new strain NA33 was amplified, sequenced and analyzed. The results showed that the complete nucleotide sequence of the S gene of NA33 strain was composed of 1 693 nucleotides with TA-rich. The S gene contained one ORF, starting at position 37 and ending at position 1 326, encoding the N protein of 429 amino acid residues, and in line with HTN-based coding. Sequence comparison of the S genes between NA33 and reference hantavirus strains showed that NA33 was more homologous to Amur-like viruses than to the Hantaan (HTN) viruses or the other hantaviruses. Phylogenetic analysis of the amino acid sequence of N proteins showed that NA33 was clustered into the group of Amur-like viruses and was more similar to Far East Russia and Jilin strains isolated from Apodemus peninsulae. The phylogenetic tree indicated a certain degree of host-dependent characteristics and geographical aggregation characteristics of hantanviruses. Furthermore, the amino acid sequence of N protein of NA33 had the conserved amino acid sites of Amur-like viruses. In conclusion, Apodemus peninsulae carried Amur-like viruses in Heilongjiang province and was an important infectious source of hemorrhagic fever with renal syndrome (HFRS).

  11. Lactobacillus casei strains isolated from cheese reduce biogenic amine accumulation in an experimental model.

    Science.gov (United States)

    Herrero-Fresno, Ana; Martínez, Noelia; Sánchez-Llana, Esther; Díaz, María; Fernández, María; Martin, Maria Cruz; Ladero, Victor; Alvarez, Miguel A

    2012-07-02

    Tyramine and histamine are the biogenic amines (BAs) most commonly found in cheese, in which they appear as a result of the microbial enzymatic decarboxylation of tyrosine and histidine respectively. Given their toxic effects, their presence in high concentrations in foods should be avoided. In this work, samples of three cheeses (Zamorano, Cabrales and Emmental) with long ripening periods, and that often have high BA concentrations, were screened for the presence of BA-degrading lactic acid bacteria (LAB). Seventeen isolates were found that were able to degrade tyramine and histamine in broth culture. All 17 isolates were identified by 16S rRNA sequencing as belonging to Lactobacillus casei. They were typed by plasmid S1-PFGE and genomic macrorestriction-PFGE analysis. Two strains (L. casei 4a and 5b) associated with high degradation rates for both BAs were selected to test how this ability might affect histamine and tyramine accumulation in a Cabrales-like mini-cheese manufacturing model. The quantification of BAs and the monitoring of the strains' growth over ripening were undertaken by RP-HPLC and qPCR respectively. Both strains were found to reduce histamine and tyramine accumulation. These two strains might be suitable for use as adjunct cultures for reducing the presence of BAs in cheese.

  12. Isolation and characteristics of Arthrobacter sp. strain CW-1 for biodegradation of PAEs

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Isolation of new bacterial strains and recognition of their metabolic activities are highly desirable for sustainability of natural ecosystems. Biodegradation of dimethyl phthalate (DMP) under anoxic conditions has been shown to occur as a series of sequential steps using strain CW-1 isolated from digested sludge of Sibao Wastewater Treatment Plant in Hangzhou, China. The microbial colony on LB medium was yellowish, 3~5 mm in diameter, convex in the center, and embedded in mucous externally.The individual cells of strain CW-1 are irregular rods, measuring (0.6~0.7)×(0.9~1.0) μm, V-shaped, with clubbed ends, Gram positive and without any filaments. 16S rDNA (1438 bp) sequence analysis showed that the strain was related to Arthrobacter sp.CW-1 and can degrade PAEs utilizing nitrate as electron acceptor, but cannot mineralize DMP completely. The degradation pathway was recommended as: dimethyl phthalate (DMP)→monomethyl phthalate (MMP)→phthalic acid (PA). DMP biodegradation was a first order reaction with degradation rate constant of 0.3033 d-1 and half-life 2.25 d. The DMP conversion to PA by CW-1 could be described by using sequential kinetic model.

  13. Characterization of Bacterial Strains Isolated from a Novel Seawater-based Retting Treatment of Hemp

    Institute of Scientific and Technical Information of China (English)

    ZHU Run-ye; CHEN Jian-yong; FENG Xin-xing; ZHANG Jian-chun

    2008-01-01

    Cultivable bacteria were isolated from seawater-based retting treatment of hemp, in which three of purified strains (SW - 1, SW - 2, and S - SW1) produced relatively high levels of pectinase activities, and also produced mannanases and xylanases.PCR - based entebacterial repetitive intergenic consensus primers (ERIC- PCR) were employed for fingerprinting DNA of the bacterial strains.The ERIC - PCR fingerprints of stains SW- 1, SW -1, and S -SW1 were found to be different, and should be further identified for each isolate.Strains SW - 1 and SW - 2 were identified as Stenotrophomnas maltophilia, while strain S - SW1 was assigned to Ochrobactrum anthropi by BIOLOG system.These two species represented rhizosphere bacterial genera, and possibly were introduced by the hemp plants.These organisms seemed potentially capable of producing pectinase and hemicellulase, and thus effectively degrading the gum substances in the seawater retting.This research could be helpful for improving a novel seawater-based retting treatment of hemp.

  14. Fruiting Body Formation of Cordyceps militaris from Multi-Ascospore Isolates and Their Single Ascospore Progeny Strains

    Science.gov (United States)

    Shrestha, Bhushan; Han, Sang-Kuk; Sung, Jae-Mo

    2012-01-01

    Interest in commercial cultivation and product development of Cordyceps species has shown a recent increase. Due to its biochemical and pharmacological effects, Cordyceps militaris, commonly known as orange caterpillar fungus, is being investigated with great interest. Cultivation of C. militaris has been practiced on a large scale in order to fulfill a demand for scientific investigation and product development. Isolates of C. militaris can be easily established from both spores and tissue. For isolation of spores, ascospores released from mature stromata are trapped in sterile medium. Multi-ascospore isolates, as well as combinations of single ascospore strains, are used for production of fruiting bodies. Progeny ascospore strains can be isolated from artificial fruiting bodies, thus, the cycle of fruiting body production can be continued for a long period of time. In this study, we examined fruiting body production from multi-ascospore isolates and their progeny strains for three generations. F1 progeny strains generally produced a larger number of fruiting bodies, compared with their mother multi-ascospore isolates; however, F2 and F3 progeny strains produced fewer fruiting bodies. Optimum preservation conditions could help to increase the vitality of the progeny strains. In order to retain the fruiting ability of the strains, further testing of various methods of preservation and different methods for isolation should be performed. PMID:22870051

  15. [Isolation of Aspergillus section Nigri strains in yerba mate in Posadas (Misiones, Argentina) and evaluation of their ochratoxigenic potential].

    Science.gov (United States)

    Castrillo, María L; Horianski, Marta A; Jerke, Gladis

    2013-01-01

    The objectives of the present work were to investigate the isolation frequency of genus Aspergillus in canchada yerba mate (YMCH) and elaborated yerba mate (YME) (Ilex paraguariensis) and the proportion of section Nigri isolates, as well as to determine ochratoxin A production by Aspergillus species section Nigri. Three hundred twenty eight Aspergillus strains from 20 samples of YMCH and 1306 Aspergillus strains from 36 samples of YME were isolated; of the total, 279 from the first group of strains and 1215 from the latter group, belonged to section Nigri. For the detection of ochratoxin A production, the strains were cultivated on Czapeck yeast extract agar and the toxin was detected by thin layer chromatography under UV light. Uniserate species predominance was observed in the 1494 strains of Aspergillus section Nigri obtained (Aspergillus japonicus var. japonicus and Aspergillus japonicus var. aculeatus), whereas none of the strains analysed showed ochratoxin A production in vitro at the detection level of the methodology employed.

  16. Potential Role of Yeast Strains Isolated from Grapes in the Production of Taurasi DOCG

    Science.gov (United States)

    Aponte, Maria; Blaiotta, Giuseppe

    2016-01-01

    Twelve samples of Aglianico grapes, collected in different locations of the Taurasi DOCG (Appellation of Controlled and Guaranteed Origin) production area were naturally fermented in sterile containers at room temperature. A total of 70 yeast cultures were isolated from countable WL agar plates: 52 in the middle of the fermentation and 18 at the end. On the basis of ITS-RFLP analysis and ITS sequencing, all cultures collected at the end of fermentations were identified as Saccharomyces (S.) cerevisiae; while, the 52 isolates, collected after 1 week, could be referred to the following species: Metschnikowia (M.) pulcherrima; Starmerella (Star.) bacillaris; Pichia (P.) kudriavzevii; Lachancea (L.) thermotolerans; Hanseniaspora (H.) uvarum; Pseudozyma (Pseud.) aphidis; S. cerevisiae. By means of Interdelta analysis, 18 different biotypes of S. cerevisiae were retrieved. All strains were characterized for ethanol production, SO2 resistance, H2S development, β-glucosidasic, esterasic and antagonistic activities. Fermentation abilities of selected strains were evaluated in micro-fermentations on Aglianico must. Within non-Saccharomyces species, some cultures showed features of technological interest. Antagonistic activity was expressed by some strains of M. pulcherrima, L. thermotolerans, P. kudriavzevii, and S. cerevisiae. Strains of M. pulcherrima showed the highest β-glucosidase activity and proved to be able to produce high concentrations of succinic acid. L. thermotolerans produced both succinic and lactic acids. The lowest amount of acetic acid was produced by M. pulcherrima and L. thermotolerans; while the highest content was recorded for H. uvarum. The strain of Star. bacillaris produced the highest amount of glycerol and was able to metabolize all fructose and malic acid. Strains of M. pulcherrima and H. uvarum showed a low fermentation power (about 4%), while, L. thermotolerans, Star. Bacillaris, and P. kudriavzevii of about 10%. Significant differences were

  17. Surveillance of Diarrheagenic Escherichia coli strains isolated from diarrhea cases from children, adults and elderly at northwest of Mexico

    Directory of Open Access Journals (Sweden)

    Adrian Canizalez-Roman

    2016-11-01

    Full Text Available Diarrheagenic Escherichia coli (DEC strains are a main cause of gastrointestinal disease in developing countries. In this study we report the epidemiologic surveillance in a four-year period (January 2011 to December 2014 of DEC strains causing acute diarrhea throughout the Sinaloa State, Mexico. DEC strains were isolated from outpatients of all ages with acute diarrhea (N=1,037. Specific DEC pathotypes were identified by PCR-amplification of genes encoding virulence factors. The adhesion phenotype and antibiotic resistance were also investigated. DEC strains were detected in 23.3% (242/1037 of cases. The most frequently DEC strain isolated was EAEC (12.2%, 126/242 followed by EPEC (5.1%, 53/242, ETEC (4.3%, 43/242 DAEC (1.4%, 15/242, STEC (0.3%, 3/242 and EIEC (0.2%, 2/242. EHEC strains were not detected. Overall DEC strains were more prevalent in children ≤ 2 years of age with EPEC strains the most common of DEC pathotypes. While 65% of EAEC strains were classified as typical variant based on the aggregative adherence to in vitro cultures of HEp-2 cells, a high proportion of EPEC strains was classified as atypical strains. EAEC, EPEC, ETEC and DAEC strains were distributed in the north, central and south regions of Sinaloa state. Among all DEC strains, >90% were resistant to at least one commonly prescribed antibiotic. Strains were commonly resistant to first-line antibiotics such as tetracycline, ampicillin and sulfamethoxazole-trimethoprim. Furthermore, more than 80% of DEC isolates were multi-drug resistant and EPEC and DAEC were the categories with major proportion of this feature. In conclusion, in nearly one out of four cases of acute diarrhea in Northwestern Mexico a multi-drug resistant DEC strain was isolated, in these cases EAEC was the most prevalent (52% pathotype.

  18. CRYSTAL STRUCTURE OF CLOSTRIDIUM BOTULINUM NEUROTOXIN SEROTYPE B.

    Energy Technology Data Exchange (ETDEWEB)

    SWAMINATHAN,S.; ESWARAMOORTHY,S.

    2001-11-19

    The toxigenic strains of Clostridium botulinum produce seven serologically distinct types of neurotoxins labeled A - G (EC 3.4.24.69), while Clostridium tetani produces tetanus neurotoxin (EC 3.4.24.68). Botulinum and tetanus neurotoxins (BoNTs and TeNT) are produced as single inactive chains of molecular mass of approximately 150 kDa. Most of these neurotoxins are released after being cleaved into two chains, a heavy chain (HI) of 100 kDa and a light chain (L) of 50 kDa held together by an interchain disulfide bond, by tissue proteinases. BoNT/E is released as a single chain but cleaved by host proteinases [1]. Clostvidium botulinum neurotoxins are extremely poisonous proteins with their LD{sub 50} for humans in the range of 0.1 - 1 ng kg{sup -1} [2]. Botulinum neurotoxins are responsible for neuroparalytic syndromes of botulism characterized by serious neurological disorders and flaccid paralysis. BoNTs block the release of acetylcholine at the neuromuscular junction causing flaccid paralysis while TeNT blocks the release of neurotransmitters like glycine and {gamma}-aminobutyric acid (GABA) in the inhibitory interneurons of the spinal cord resulting in spastic paralysis. In spite of different clinical symptoms, their aetiological agents intoxicate neuronal cells in the same way and these toxins have similar structural organization [3].

  19. Isolation and Characterization of Campylobacter Strains from Diarrheal Patients in Central and Suburban Bangkok, Thailand.

    Science.gov (United States)

    Samosornsuk, Worada; Asakura, Masahiro; Yoshida, Emi; Taguchi, Takashi; Eampokalap, Bunchuay; Chaicumpa, Wanpen; Yamasaki, Shinji

    2015-01-01

    Campylobacter-induced diarrhea is increasingly recognized worldwide. However, little information is available regarding the Campylobacter strains associated with diarrheal patients in Thailand. In this study, we attempted to isolate Campylobacter strains from diarrheal patients in Thailand and to characterize the species using a cytolethal distending toxin (cdt) gene-based C. jejuni, C. coli, and C. fetus-specific multiplex PCR assay. Campylobacter species were also confirmed using 16S rRNA gene sequencing and hipO gene detection. From 2,500 diarrheal stool specimens, 76 Campylobacter-like organisms were isolated and identified via conventional culture methods. Among these 76 organisms, 73 were identified as Campylobacter species (43 C. jejuni, 29 C. coli, and 1 C. fetus) via multiplex PCR, whereas 3 remained unidentified. Two Campylobacter-like organisms yielded 2 amplicons corresponding to cdt genes from C. jejuni and C. coli. Subsequently, C. jejuni and C. coli were reisolated from each sample. The third isolate was identified as C. hyointestinalis via 16S rRNA gene sequencing. To our knowledge, this is the first report on the isolation of C. hyointestinalis from a diarrheal patient in Thailand. These data indicate that C. jejuni (58%) and C. coli (40%) are prevalent among diarrheal patients in Thailand.

  20. Molecular characterization of Campylobacter lanienae strains isolated from food-producing animals.

    Science.gov (United States)

    Schweitzer, Nóra; Damjanova, Ivelina; Kaszanyitzky, Eva; Ursu, Krisztina; Samu, Péterné; Tóth, Adám György; Varga, János; Dán, Adám

    2011-05-01

    During 2008 and 2009, within the framework of the Hungarian monitoring program of antibiotic resistance of zoonotic agents from food-producing animals, a significant number (43 strains) of Campylobacter lanienae were detected for the first time in Hungary. The isolates were genotyped using partial 16S rRNA gene sequencing and pulsed-field gel electrophoresis using three different restriction enzymes. The antimicrobial resistance of the isolates was determined by microtiter broth dilution. C. lanienae isolation was successful only from swine but not from other animal species. According to phylogenetic analysis, clustering of the isolates shows the same extensive genetic diversity as other Campylobacter species. Sequence analysis of the partial 16S rRNA gene showed that additional variations exist in variable regions Vc2 and Vc6. SmaI restriction enzyme proved to be the most efficient for pulsed-field gel electrophoresis analysis of C. lanienae. A significant tetracycline resistance (60.9%) and the presence of erythromycin-, enrofloxacin-, and multiresistant C. lanienae strains were found. Although the pathogenic potential of C. lanienae in humans is currently unknown, this study demonstrates that C. lanieanae is common in pigs in the country, provides further details on the genotypic and phenotypic properties of C. lanienae, and offers a genotyping method for use in source tracing.

  1. Enterotoxigenic Genes in strains of Staphylococcus spp., isolated from cheese made in Pamplona-Colombia

    Directory of Open Access Journals (Sweden)

    Fanny Herrera A.

    2015-01-01

    Full Text Available Objective. To determine the incidence of coagulase-positive strains of enterotoxigenic Staphylococcus in doble crema (double cream cheese samples produced in Pamplona. Materials and methods. Bacterial isolation was performed following the routine method for coagulase positive Staphylococcus provided by the Colombian Technical Standard 4779, by using Baird Parker medium with confirmation of typical colonies by performing the coagulase test. Detection of genes for principal enterotoxins was done by PCR. Results. The prevalence of coagulase positive Staphylococcus in cheese samples was 31%, with 27% of the samples failing to meet the requirements of the NTC 750. In 24.6% of the studied isolates, genes for enterotoxin production were detected. The presence, in the isolated strains, of genes for SEB, SEA and SED was 18.5%, 4.6% and 3.0%, respectively. Conclusions. The significant presence of enterotoxigenic genes found in the isolates obtained from samples of double cream cheese made in Pamplona, suggests an important hazard to the health of consumers.

  2. PENICILLIUM STRAINS ISOLATED FROM GRAPES GROWN IN THE CENTRAL SLOVAK WINE REGION

    Directory of Open Access Journals (Sweden)

    Soňa Felšöciová

    2014-02-01

    Full Text Available A total of 10 wine producing grapes were collected from the Central Slovak region in 2011 and 2012, which involved 7 vineyards. Mycological analysis was carried out for the detection of fungi using standard media with focus on genera Penicillium. The exogenous mycobiota was determined by the method of direct placing of berry samples on agar plates and the endogenous mycobiota by the same method but the berry samples were first superficially sterilized. The resulting Penicillia were identified morphologically. The potentially toxigenic isolates were analysed for mycotoxin production by TLC method. In general representatives of Penicillium genus were isolated with higher frequency from the surface of the berries than from their interior. In regard to species Penicillium chrysogenum seems to be the most wide-spread in wine grape berries from the Central Slovak wine growing area because was detected more frequently than other species. In total, 546 fungal isolates belonging to 12 Penicillium species. Overall, two species were dominant: P. chrysogenum and P. expansum. Four potentially toxigenic species isolated from endogenous mycobiota were tested for their toxigenic ability. Out of 10 strains, all of them produced at least one mycotoxin. Of all 16 potentially toxigenic strains from exogenous mycobiota, 88% produced at least one mycotoxin. Therefore study concluded, that wine producing grapes seems to be risk products.

  3. Metabolic functions of Pseudomonas fluorescens strains from Populus deltoides depend on rhizosphere or endosphere isolation compartment

    Directory of Open Access Journals (Sweden)

    Collin M Timm

    2015-10-01

    Full Text Available The bacterial microbiota of plants is diverse, with 1,000s of operational taxonomic units (OTUs associated with any individual plant. In this work we investigate the differences between 19 sequenced Pseudomonas fluorescens strains, isolated from Populus deltoides rhizosphere and endosphere and which represent a single OTU, using phenotypic analysis, comparative genomics, and metabolic models. While no traits were exclusive to either endosphere or rhizosphere P. fluorescens isolates, multiple pathways relevant for plant-bacterial interactions are enriched in endosphere isolate genomes. Further, growth phenotypes such as phosphate solubilization, protease activity, denitrification and root growth promotion are biased towards endosphere isolates. Endosphere isolates have significantly more metabolic pathways for plant signaling compounds and an increased metabolic range that includes utilization of energy rich nucleotides and sugars, consistent with endosphere colonization. Rhizosphere P. fluorescens have fewer pathways representative of plant-bacterial interactions but show metabolic bias towards chemical substrates often found in root exudates. This work reveals the diverse functions that may contribute to colonization of the endosphere by bacteria and are enriched among closely related isolates.

  4. Characterization of Insecticidal Genes of Bacillus thuringiensis Strains Isolated from Arid Environments.

    Science.gov (United States)

    Abulreesh, Hussein H; Osman, Gamal E H; Assaeedi, Abdulrahman S A

    2012-09-01

    This study aimed at characterizing the insecticidal genes of eight Bacillus thuringiensis isolates that were recovered from the local environment of western Saudi Arabia. The screening for the presence of lepidopteran-specific cry1A family and vip3A genes, dipteran-specific cry4 family and coleopteran-specific cry3A, vip1A and vip2A genes, was carried out by PCR. All eight isolates produced PCR products that confirmed the presence of cry1Aa, cry1Ab, cry1Ac, cry4A, cry4B genes, but not cry3A, vip1A and vip2A genes. However, three isolates only were found to carry vip3A genes as revealed by PCR. The observation of cry1 and cry4 genes suggests that these eight isolates may have dual activity against Lepidoptera and Diptera species, while three isolates possessed vip3 genes in addition to cry1 and cry4 which suggests that these three isolates have toxic crystals and vegetative proteins. The results of this study are interesting in the sense that they may help developing new strategies for controlling insects of economic and medical importance in Saudi Arabia, using B. thuringiensis strains that naturally exist in the local environment instead of the current control strategies that are based solely on chemical insecticides.

  5. Isolation and identification of oedogonium species and strains for biomass applications.

    Directory of Open Access Journals (Sweden)

    Rebecca J Lawton

    Full Text Available Freshwater macroalgae from the genus Oedogonium have recently been targeted for biomass applications; however, strains of Oedogonium for domestication have not yet been identified. Therefore, the objective of this study was to compare the performance of isolates of Oedogonium collected from multiple geographic locations under varying environmental conditions. We collected and identified wild-type isolates of Oedogonium from three geographic locations in Eastern Australia, then measured the growth of these isolates under a range of temperature treatments corresponding to ambient conditions in each geographic location. Our sampling identified 11 isolates of Oedogonium that could be successfully maintained under culture conditions. It was not possible to identify most isolates to species level using DNA barcoding techniques or taxonomic keys. However, there were considerable genetic and morphological differences between isolates, strongly supporting each being an identifiable species. Specific growth rates of species were high (>26% day-1 under 7 of the 9 temperature treatments (average tested temperature range: 20.9-27.7°C. However, the variable growth rates of species under lower temperature treatments demonstrated that some were better able to tolerate lower temperatures. There was evidence for local adaptation under lower temperature treatments (winter conditions, but not under higher temperature treatments (summer conditions. The high growth rates we recorded across multiple temperature treatments for the majority of species confirm the suitability of this diverse genus for biomass applications and the domestication of Oedogonium.

  6. Isolation and identification of oedogonium species and strains for biomass applications.

    Science.gov (United States)

    Lawton, Rebecca J; de Nys, Rocky; Skinner, Stephen; Paul, Nicholas A

    2014-01-01

    Freshwater macroalgae from the genus Oedogonium have recently been targeted for biomass applications; however, strains of Oedogonium for domestication have not yet been identified. Therefore, the objective of this study was to compare the performance of isolates of Oedogonium collected from multiple geographic locations under varying environmental conditions. We collected and identified wild-type isolates of Oedogonium from three geographic locations in Eastern Australia, then measured the growth of these isolates under a range of temperature treatments corresponding to ambient conditions in each geographic location. Our sampling identified 11 isolates of Oedogonium that could be successfully maintained under culture conditions. It was not possible to identify most isolates to species level using DNA barcoding techniques or taxonomic keys. However, there were considerable genetic and morphological differences between isolates, strongly supporting each being an identifiable species. Specific growth rates of species were high (>26% day-1) under 7 of the 9 temperature treatments (average tested temperature range: 20.9-27.7°C). However, the variable growth rates of species under lower temperature treatments demonstrated that some were better able to tolerate lower temperatures. There was evidence for local adaptation under lower temperature treatments (winter conditions), but not under higher temperature treatments (summer conditions). The high growth rates we recorded across multiple temperature treatments for the majority of species confirm the suitability of this diverse genus for biomass applications and the domestication of Oedogonium.

  7. Antimicrobial Activity of an Alcaligenes faecalis Strain Isolated from Oil Contaminated Soil

    Directory of Open Access Journals (Sweden)

    Yaghoobi Avini, M. (MSc

    2015-01-01

    Full Text Available Background and Objective: The bacteria living in the specific ecological conditions are among the most promising antimicrobial producers. This study aimed at isolating antimicrobial producing bacteria from soils contaminated with crude oil. Material and Methods: the samples were obtained from crude oil contaminated soils around Dezful located in Khuzestan province, Iran, and antimicrobial producing bacteria were isolated using disc diffusion and cross streak culture. Then, the best bacterium was selected and its antimicrobial potency was studied against indicator microorganisms. The isolate was also characterized based on biochemical properties and phylogenetic analysis. Results: based on the results, the highest antimicrobial activity of isolated bacterium was related to Candida albicans, Aspergillus niger, Bacillus subtilis, E. coli and Klebsiella pneumonia. An intermediate effect was determined against Serratia marcesens and Staphylococcus aureus, whereas no effect was observed against three strains of Enterococcus. Using biochemical characteristics and phenotypic traits, the isolate was identified as Alcaligenes faecalis. Conclusion: given that the isolate has broad spectrum activity against a various range of microorganisms and in comparison with some antimicrobial compounds produced by other Alcaligenes species, it seems the novelty of this antimicrobial compound.

  8. Genetic Analysis of Vibrio parahaemolyticus O3:K6 Strains That Have Been Isolated in Mexico Since 1998

    Science.gov (United States)

    Licea-Navarro, Alexei Fedorovish; Revilla-Castellanos, Valeria Jeanette; Wong-Chang, Irma; González-Sánchez, Ricardo

    2017-01-01

    Vibrio parahaemolyticus is an important human pathogen that has been isolated worldwide from clinical cases, most of which have been associated with seafood consumption. Environmental and clinical toxigenic strains of V. parahaemolyticus that were isolated in Mexico from 1998 to 2012, including those from the only outbreak that has been reported in this country, were characterized genetically to assess the presence of the O3:K6 pandemic clone, and their genetic relationship to strains that are related to the pandemic clonal complex (CC3). Pathogenic tdh+ and tdh+/trh+ strains were analyzed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Also, the entire genome of a Mexican O3:K6 strain was sequenced. Most of the strains were tdh/ORF8-positive and corresponded to the O3:K6 serotype. By PFGE and MLST, there was very close genetic relationship between ORF8/O3:K6 strains, and very high genetic diversities from non-pandemic strains. The genetic relationship is very close among O3:K6 strains that were isolated in Mexico and sequences that were available for strains in the CC3, based on the PubMLST database. The whole-genome sequence of CICESE-170 strain had high similarity with that of the reference RIMD 2210633 strain, and harbored 7 pathogenicity islands, including the 4 that denote O3:K6 pandemic strains. These results indicate that pandemic strains that have been isolated in Mexico show very close genetic relationship among them and with those isolated worldwide. PMID:28099500

  9. Antagonistic effect of Lactobacillus strains against gas-producing coliforms isolated from colicky infants

    Directory of Open Access Journals (Sweden)

    Oggero Roberto

    2011-06-01

    Full Text Available Abstract Background Infantile colic is a common disturb within the first 3 months of life, nevertheless the pathogenesis is incompletely understood and treatment remains an open issue. Intestinal gas production is thought to be one of the causes of abdominal discomfort in infants suffering from colic. However, data about the role of the amount of gas produced by infants' colonic microbiota and the correlation with the onset of colic symptoms are scanty. The benefit of supplementation with lactobacilli been recently reported but the mechanisms by which they exert their effects have not yet been fully defined. This study was performed to evaluate the interaction between Lactobacillus spp. strains and gas-forming coliforms isolated from stools of colicky infants. Results Strains of coliforms were isolated from stools of 45 colicky and 42 control breastfed infants in McConkey Agar and identified using PCR with species-specific primers, and the BBL™ Enterotube™ II system for Enterobacteriaceae. Gas-forming capability of coliforms was assessed in liquid cultures containing lactose as sole carbon source. The average count of total coliforms in colicky infants was significantly higher than controls: 5.98 (2.00-8.76 log10 vs 3.90 (2.50-7.10 CFU/g of faeces (p = 0.015. The following strains were identified: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae and Enterococcus faecalis. Then, 27 Lactobacillus strains were tested for their antagonistic effect against coliforms both by halo-forming method and in liquid co-cultures. Lactobacillus delbrueckii subsp.delbrueckii DSM 20074 and L. plantarum MB 456 were able to inhibit all coliforms strains (halo-forming method, also in liquid co-cultures, thus demonstrating an antagonistic activity. Conclusions This study shows that two out of 27 strains of Lactobacillus examined possess an antimicrobial effect against six species of gas-forming coliforms

  10. Technological properties and probiotic potential of Lactobacillus fermentum strains isolated from West African fermented millet dough

    DEFF Research Database (Denmark)

    Owusu-Kwarteng, James; Tano-Debrah, Kwaku; Akabanda, Fortune;

    2015-01-01

    the isolation of indigenous strains from traditional fermented products to be used as functional starter cultures. These functional starter cultures possess inherent functional characteristics and can contribute to food quality and safety by offering one or more organoleptic, nutritional, technological...... activity was observed towards Listeria monocytogenes and Staphylococcus aureus but not E. coli and Salmonella enteritidis. Lactobacillus fermentum strains were generally susceptible to antibiotics except 6 strains which showed resistance towards streptomycin, gentamicin and kanamycin. CONCLUSION: In vitro...... determination of technological and probiotic properties have shown strain specific difference among L. fermentum strains isolated from fermented millet dough. Sixteen (16) L. fermentum strains have been shown to possess desirable technological and probiotic characteristics in vitro. These strains are therefore...

  11. Development of a Potential Probiotic Fresh Cheese Using Two Lactobacillus salivarius Strains Isolated from Human Milk

    Directory of Open Access Journals (Sweden)

    Nivia Cárdenas

    2014-01-01

    Full Text Available Cheeses have been proposed as a good alternative to other fermented milk products for the delivery of probiotic bacteria to the consumer. The objective of this study was to assess the survival of two Lactobacillus salivarius strains (CECT5713 and PS2 isolated from human milk during production and storage of fresh cheese for 28 days at 4°C. The effect of such strains on the volatile compounds profile, texture, and other sensorial properties, including an overall consumer acceptance, was also investigated. Both L. salivarius strains remained viable in the cheeses throughout the storage period and a significant reduction in their viable counts was only observed after 21 days. Globally, the addition of the L. salivarius strains did not change significantly neither the chemical composition of the cheese nor texture parameters after the storage period, although cheeses manufactured with L. salivarius CECT5713 presented significantly higher values of hardness. A total of 59 volatile compounds were identified in the headspace of experimental cheeses, and some L. salivarius-associated differences could be identified. All cheeses presented good results of acceptance after the sensory evaluation. Consequently, our results indicated that fresh cheese can be a good vehicle for the two L. salivarius strains analyzed in this study.

  12. Bacillus sp. strain DJ-1, potent arsenic hypertolerant bacterium isolated from the industrial effluent of India.

    Science.gov (United States)

    Joshi, Dhaval N; Flora, S J S; Kalia, Kiran

    2009-07-30

    Arsenic hypertolerant bacterial cells were isolated from the common industrial effluent treatment plant, Vapi, India. Strain DJ-1 sustaining 400 mM, As (V) out of 16 bacterial strains was identified as Bacillus sp. strain DJ-1 through 16S rRNA ribotyping. The maximum arsenic accumulation of 9.8+/-0.5 mg g(-1) (dry weight) was observed during stationary phase of growth. Intracellular compartmentalization has shown 80% of arsenic accumulation in cytoplasm. The lack of arsC gene and arsenate reductase activity indicated that Bacillus sp. strain DJ-1 may lack classical ars operon and detoxification may be mediated through some novel mechanism. The arsenite binding protein was purified by affinity chromatography and characterized as DNA protection during starvation (DPS) protein by electrospray ionization mass spectrometry. The induction of DPS showed the adaptation of bacteria in arsenic stress condition and/or in detoxification mechanism, relies on its ability to bind with arsenic. These results indicate the hypertolerance with higher intracellular accumulation of arsenic by Bacillus sp. strain DJ-1, which could be mediated by DPS protein thus signifying this organism is a potential candidate for the removal of arsenic from industrial wastewater, which needs further study.

  13. Inhibition of mycotoxin-producing fungi by Bacillus strains isolated from fish intestines.

    Science.gov (United States)

    Veras, Flávio Fonseca; Correa, Ana Paula Folmer; Welke, Juliane Elisa; Brandelli, Adriano

    2016-12-05

    Bacillus strains isolated from the aquatic environment of the Brazilian Amazon region were tested for their activity against mycotoxigenic fungi. All tested bacteria showed antifungal activity, inhibiting at least 7 indicator fungi. Four Bacillus strains showing promising antifungal results were subsequently evaluated for their activity in reducing mycelial growth rate, sporulation, spore germination percentage, and mycotoxin production. Bacillus sp. P1 and Bacillus sp. P11 had a remarkable antifungal effect on toxigenic fungi. Washed bacterial cell suspension of strains P1 and P11 (10(7)CFU/ml) reduced by >70% the fungal colony diameters, including a complete inhibition of ochratoxin A (OTA) producing Aspergillus spp. Significant reduction of growth rate, sporulation and spore germination were also observed. The bacteria influenced the production of mycotoxins, causing a reduction around 99 and 97% in AFB1 and OTA concentration, respectively. Chromatographic analysis revealed the presence of lipopeptides (iturin A and surfactin isomers) in butanol extracts of cell-free supernatants and cell pellets of strains P1 and P11. Furthermore, antifungal activity of these extracts was confirmed against A. flavus A12 and A. carbonarius ITAL293, producers of AFB1 and OTA, respectively. These bacterial strains could be promising biocontrol agents against toxigenic fungi.

  14. Hexavalent Chromium Removal by a Paecilomyces sp. Fungal Strain Isolated from Environment

    Directory of Open Access Journals (Sweden)

    Juan F. Cárdenas-González

    2010-01-01

    Full Text Available A resistant and capable fungal strain in removing hexavalent chromium was isolated from an environment near of Chemical Science Faculty, located in the city of San Luis Potosí, Mexico. The strain was identified as Paecilomyces sp., by macro- and microscopic characteristics. Strain resistance of the strain to high Cr (VI concentrations and its ability to reduce chromium were studied. When it was incubated in minimal medium with glucose, another inexpensive commercial carbon source like unrefined and brown sugar or glycerol, in the presence of 50 mg/L of Cr (VI, the strain caused complete disappearance of Cr (VI, with the concomitant production of Cr (III in the growth medium after 7 days of incubation, at 28∘C, pH 4.0, 100 rpm, and an inoculum of 38 mg of dry weight. Decrease of Cr (VI levels from industrial wastes was also induced by Paecilomyces biomass. These results indicate that reducing capacity of chromate resistant filamentous fungus Cr (VI could be useful for the removal of Cr (VI pollution.

  15. Production and characterization of L-fucose dehydrogenase from newly isolated Acinetobacter sp. strain SA-134.

    Science.gov (United States)

    Ohshiro, Takashi; Morita, Noriyuki

    2014-01-01

    Microorganisms producing L-fucose dehydrogenase were screened from soil samples, and one of the isolated bacterial strains SA-134 was identified as Acinetobacter sp. by 16S rDNA gene analysis. The strain grew well utilizing L-fucose as a sole source of carbon, but all other monosaccharides tested such as D-glucose and D-arabinose did not support the growth of the strain in the absence of L-fucose. D-Arabinose inhibited the growth even in the culture medium containing L-fucose. Although the strain grew on some organic acids and amino acids such as citric acid and L-alanine as sole sources of carbon, the enzyme was produced only in the presence of L-fucose. The fucose dehydrogenase was purified to apparently homogeneity from the strain, and the native enzyme was a monomer of 25 kD. L-Fucose and D-arabinose were good substrates for the enzyme, but L-galactose was a poor substrate. The enzyme acted on both NAD(+) and NADP(+) in the similar manner.

  16. Measurement of antimicrobial activity of isolated bacteria from the Caspian sea and molecular identification of strains with antimicrobial effect

    Directory of Open Access Journals (Sweden)

    Sajad Harounabadi

    2015-12-01

    Full Text Available Introduction: Easy access and wide use of antimicrobial compounds led to the emergence of resistance among microorganisms. Therefore, screening and identifying antimicrobial compound with high effect of microorganisms in different environments is necessary and vital . Using microorganisms for biological aims change them to an important tool to control pathogens. Streptomyces griseus is one of them. The aim of this study is isolation of marine bacteria with antimicrobial effect against gram positive and negative bacteria. Finally, molecular identification of strains with antimicrobial activity. Materials and methods: In this study, 162 strains were isolated from the Caspian Sea .The strains were cultured on special medium and finally antimicrobial activity on references strains as measured. Among them four strains with remarkable antimicrobial activity were identified and selected. The strains were subjected to 16S rDNA PCR sequencing. The strains were submitted to NCBI as new Streptomyces griseus strains. Results: Among 162 strains, 4 strains had the most antimicrobial activity. The result showed, the strains were the most effective on Bacillus subtilis and Staphylococcus aureus (Gram positive bacteria and the least effect were observed on Escherichia coli and Pseudomonas aeruginosa (Gram negative bacteria. After sequencing, the strains were classified to sterptomyces griseus genu. Discussion and conclusion: In this study, 4 strains with antimicrobial activity were identified. According to the strength of these bacteria for controlling pathogenic bacteria resistant to antibiotic, we can have more pure microorganisms in optimized and controlled conditions for using in pharmaceutical industries and also for the treatment of dangerous pathogenic bacteria.

  17. Independent evolution of neurotoxin and flagellar genetic loci in proteolytic Clostridium botulinum

    Directory of Open Access Journals (Sweden)

    Twine Susan M

    2009-03-01

    Full Text Available Abstract Background Proteolytic Clostridium botulinum is the causative agent of botulism, a severe neuroparalytic illness. Given the severity of botulism, surprisingly little is known of the population structure, biology, phylogeny or evolution of C. botulinum. The recent determination of the genome sequence of C. botulinum has allowed comparative genomic indexing using a DNA microarray. Results Whole genome microarray analysis revealed that 63% of the coding sequences (CDSs present in reference strain ATCC 3502 were common to all 61 widely-representative strains of proteolytic C. botulinum and the closely related C. sporogenes tested. This indicates a relatively stable genome. There was, however, evidence for recombination and genetic exchange, in particular within the neurotoxin gene and cluster (including transfer of neurotoxin genes to C. sporogenes, and the flagellar glycosylation island (FGI. These two loci appear to have evolved independently from each other, and from the remainder of the genetic complement. A number of strains were atypical; for example, while 10 out of 14 strains that formed type A1 toxin gave almost identical profiles in whole genome, neurotoxin cluster and FGI analyses, the other four strains showed divergent properties. Furthermore, a new neurotoxin sub-type (A5 has been discovered in strains from heroin-associated wound botulism cases. For the first time, differences in glycosylation profiles of the flagella could be linked to differences in the gene content of the FGI. Conclusion Proteolytic C. botulinum has a stable genome backbone containing specific regions of genetic heterogeneity. These include the neurotoxin gene cluster and the FGI, each having evolved independently of each other and the remainder of the genetic complement. Analysis of these genetic components provides a high degree of discrimination of strains of proteolytic C. botulinum, and is suitable for clinical and forensic investigations of botulism

  18. Strain Diversity of Pseudomonas fluorescens Group with Potential Blue Pigment Phenotype Isolated from Dairy Products.

    Science.gov (United States)

    Chierici, Margherita; Picozzi, Claudia; La Spina, Marisa Grazia; Orsi, Carla; Vigentini, Ileana; Zambrini, Vittorio; Foschino, Roberto

    2016-08-01

    The blue discoloration in Mozzarella cheese comes from bacterial spoilage due to contamination with Pseudomonas. Fourteen Pseudomonas fluorescens strains from international collections and 55 new isolates of dominant bacterial populations from spoiled fresh cheese samples were examined to assess genotypic and phenotypic strain diversity. Isolates were identified by 16S rRNA gene sequencing and tested for the production of the blue pigment at various temperatures on Mascarpone agar and in Mozzarella preserving fluid (the salty water in which the cheese is conserved, which becomes enriched by cheese minerals and peptides during storage). Pulsed-field gel electrophoresis analysis after treatment with the endonuclease SpeI separated the isolates into 42 genotypes at a similarity level of 80%. Based on the pulsotype clustering, 12 representative strains producing the blue discoloration were chosen for the multilocus sequence typing targeting the gyrB, glnS, ileS, nuoD, recA, rpoB, and rpoD genes. Four new sequence typing profiles were discovered, and the concatenated sequences of the investigated loci grouped the tested strains into the so-called ''blue branch'' of the P. fluorescens phylogenetic tree, confirming the linkage between pigment production and a specific genomic cluster. Growth temperature affected pigment production; the blue discoloration appeared at 4 and 14°C but not at 30°C. Similarly, the carbon source influenced the phenomenon; the blue phenotype was generated in the presence of glucose but not in the presence of galactose, sodium succinate, sodium citrate, or sodium lactate.

  19. [Isolation and Identification of Petroleum Degradation Bacteria and Interspecific Interactions Among Four Bacillus Strains].

    Science.gov (United States)

    Wang, Jia-nan; Shi, Yan-yun; Zheng, Li-yan; Wang, Zhe; Cai, Zhang; Liu, Jie

    2015-06-01

    Six petroleum-degrading strains were isolated from oil-contaminated soil at Dagang oil field and oil sewage on Bohai offshore drilling platform in Tianjin using enrichment culture and isolation method. The physiological biochemical test together with 16S rDNA sequencing analysis indicated that they belonged to Bacillus (S1, S2, S3, S4), Pseudomonas (W1) and Ochrobactrum (W2), respectively. The strain S3 had the maximum degradation rate of alkane (41.3%) and aromatic hydrocarbon (30.9%) among all isolated strains showing the better degradation efficiency by endogenous bacteria when compared to that by the exogenous bacteria. The four Bacillus strains were used to construct microbiome, thereafter subjected to petroleum degradation efficiency test and analyzed. The results showed that microbiome F3 consisting of S1 and S4 had the maximum degradation rates of alkane (50.5%) and aromatic hydrocarbon (54.0%), which were 69.9% and 156.1% higher than those by single bacterium, respectively. Furthermore, they were 22.1% and 74.6% respectively higher than those by the most optimal degradation bacterium S3. Microbiome F4 consisting of S2 and S3 had the minimum degradation rates of alkane (18.5%) and aromatic hydrocarbon (18.9%) which were 55.3% and 39.0% lower than the degradation rates of single bacterium, respectively. The results also demonstrated that there were both microbial synergy promotion and antagonism inhibition among bacteria of the same genus in the petroleum degradation period. Bacteria with close affinity in Bacillus genus displayed mainly promoted petroleum degradation effect.

  20. Genome Analysis of a New Rhodothermaceae Strain Isolated from a Hot Spring

    Directory of Open Access Journals (Sweden)

    Kian Mau Goh

    2016-07-01

    Full Text Available A bacterial strain, designated RA, was isolated from water sample of a hot spring on Langkawi Island of Malaysia using marine agar. Strain RA is an aerophilic and thermophilic microorganism that grows optimally at 50–60°C and is capable of growing in marine broth containing 1–10% (w/v NaCl. 16S rRNA gene sequence analysis demonstrated that this strain is most closely related (< 90% sequence identity to Rhodothermaceae, which currently comprises of six genera: Rhodothermus (two species, Salinibacter (three species, Salisaeta (one species, Rubricoccus (one species, Rubrivirga (one species, and Longimonas (one species. Notably, analysis of average nucleotide identity values indicated that strain RA may represent the first member of a novel genus of Rhodothermaceae. The draft genome of strain RA is 4,616,094 bp with 3,630 protein-coding gene sequences. Its GC content is 68.3%, which is higher than that of most other genomes of Rhodothermaceae. Strain RA has genes for sulfate permease and arylsulfatase to withstand the high sulfur and sulfate contents of the hot spring. Putative genes encoding proteins involved in adaptation to osmotic stress were identified which encode proteins namely Na+/H+ antiporters, a sodium/solute symporter, a sodium/glutamate symporter, trehalose synthase, malto-oligosyltrehalose synthase, choline-sulfatase, potassium uptake proteins (TrkA and TrkH, osmotically inducible protein C, and the K+ channel histidine kinase KdpD. Furthermore, genome description of strain RA and comparative genome studies in relation to other related genera provide an overview of the uniqueness of this bacterium.

  1. Molecular epidemiology of Trichophyton tonsurans strains isolated in Japan between 2006 and 2010 and their susceptibility to oral antimycotics.

    Science.gov (United States)

    Anzawa, Kazushi; Mochizuki, Takashi; Nishibu, Akiko; Ishizaki, Hiroshi; Kamei, Katsuhiko; Takahashi, Yoko; Fujihiro, Machiko; Shinoda, Hidekazu

    2011-01-01

    Trichophyton tonsurans has been isolated among judo practitioners, wrestlers, and sumo wrestlers during an epidemic of tinea corporis and tinea capitis in Japan. A previous study using restriction fragment length polymorphism (RFLP) analysis of the non-transcribed spacer (NTS) region of the ribosomal RNA gene revealed that different sources for the causative fungus in epidemics among judo practitioners and among wrestlers. Many different fungal strains have since been isolated from practitioners of these sports. The present study evaluated fungal characteristics of strains newly isolated between July 2006 and December 2010 using this molecular method. PCR-RFLP analysis using MvaI and AvaI was performed on 263 strains, composed of 186 isolates from judo practitioners, 32 from wrestlers, 30 from sumo wrestlers, 5 from other sports, 7 from family members or friends of the sports practitioner patients, and 3 from sporadic (non-epidemic) cases. Four molecular types, NTS I, II, III, and VII were detected. Of these, NTS I was the most predominant, occurring in 243 of 263 strains (92.4%). All of the 30 strains isolated from sumo wrestlers were classified as NTS I, suggesting that the epidemic among sumo wrestlers originated from an earlier epidemic among judo practitioners. Thirteen strains were classified as NTS II; all were related to wrestling and were isolated mainly from Chubu and Kansai areas in the central part of Honshu island. NTS III was detected in 6 strains, and one strain classified as NTS VII was isolated from a sporadic case of tinea capitis in a Peruvian immigrant. The minimum inhibitory concentrations (MICs) of terbinafine, itraconazole, fluconazole, and griseofulvin on 10 strains of NTS I and NTS II and 4 strains of NTS III were examined; there were no differences in MIC between these molecular types.

  2. Comparison of Haemophilus parasuis reference strains and field isolates by using random amplified polymorphic DNA and protein profiles

    Directory of Open Access Journals (Sweden)

    Zehr Emilie S

    2012-06-01

    Full Text Available Abstract Background Haemophilus parasuis is the causative agent of Glässer’s disease and is a pathogen of swine in high-health status herds. Reports on serotyping of field strains from outbreaks describe that approximately 30% of them are nontypeable and therefore cannot be traced. Molecular typing methods have been used as alternatives to serotyping. This study was done to compare random amplified polymorphic DNA (RAPD profiles and whole cell protein (WCP lysate profiles as methods for distinguishing H. parasuis reference strains and field isolates. Results The DNA and WCP lysate profiles of 15 reference strains and 31 field isolates of H. parasuis were analyzed using the Dice and neighbor joining algorithms. The results revealed unique and reproducible DNA and protein profiles among the reference strains and field isolates studied. Simpson’s index of diversity showed significant discrimination between isolates when three 10mer primers were combined for the RAPD method and also when both the RAPD and WCP lysate typing methods were combined. Conclusions The RAPD profiles seen among the reference strains and field isolates did not appear to change over time which may reflect a lack of DNA mutations in the genes of the samples. The recent field isolates had different WCP lysate profiles than the reference strains, possibly because the number of passages of the type strains may affect their protein expression.

  3. Selected Lactobacillus strains isolated from sugary and milk kefir reduce Salmonella infection of epithelial cells in vitro.

    Science.gov (United States)

    Zavala, L; Golowczyc, M A; van Hoorde, K; Medrano, M; Huys, G; Vandamme, P; Abraham, A G

    2016-09-01

    The isolation of potentially probiotic strains and the subsequent study of their properties are very important steps to gain insight in the health benefits ascribed to sugary and milk kefir. The aim of the present study was to characterise fifteen Lactobacillus strains isolated from these beverages by determining some surface properties and their ability to antagonise enterocyte cell damage after Salmonella infection in vitro. Lactobacillus surface properties were determined by hydrophobicity, autoaggregation, and coaggregation assays with Salmonella. In addition, lactobacilli adhesion to Caco-2/TC-7 cells and the effect on Salmonella invasion were evaluated. Finally, the disassembly of F-actin cytoskeleton on intestinal epithelial cells was assayed in vitro when Salmonella infection was performed in the presence of selected Lactobacillus strains. Ten out of the 15 strains showed a high adhesion capacity to Caco-2/TC-7 cells. Most of the strains were hydrophilic and non-autoaggregating. Strains isolated from sugary kefir were non-coaggregating with Salmonella, while strains Lactobacillus paracasei CIDCA 83120, 83121, 83123, 83124, 8339, 83102 isolated from milk kefir were able to coaggregate after 1 h. L. paracasei CIDCA 8339 and Lactobacillus kefiri CIDCA 83102 were able to diminish Salmonella invasion to the enterocytes. An antagonistic effect on cytoskeleton disruption elicited by the pathogen was also demonstrated. Our results suggest that both strains isolated from milk kefir could be considered as appropriate probiotic candidates.

  4. Isolation and Identification of a new Bacillus cereus strain and Characterization of its Neopullulanase

    Directory of Open Access Journals (Sweden)

    Soheila Davaeifar

    2015-03-01

    Full Text Available Identification and use of more efficient enzymes in the food and pharmaceutical industries is the focus of many researchers. The aim of this study was to search for a new bacterial strain capable of producing high levels of pullulanase applicable to biotechnology, the starch bioprocessing and food industries. A new pullulan hydrolyzing Bacillus strain was isolated and designated SDK2. Morphological and biochemical tests identified the strain as a putative Bacillus cereus strain, which was further characterized and confirmed through 16s rRNA sequencing, and was submitted to GeneBank, under the accession number FR6864500. Quantative analysis of the strain’s pullulanase activity was carried out by the Dintrosalicyclic (DNS acid-based assay. Thin layer chromatography (TLC of the culture supernatant, identified the extracellular pullulanase as neopullulanase. Effects of temperature and pH on pullulanase activity were also studied. The optimum conditions for enzyme activity, as represented by 60o C and a pH of 7, resulted in an activity of 13.43 U/ml, which is much higher than some of the previously reported activities. However, growth of B. cereus SDK2 was also observed at a pH range of 5 to 10, and temperatures of 30 oC to 50 oC. The effect of metal ions and reagents, such as Mg+2, Ca+2, Zn+2, Cu+2, Fe+2, Ni+2 on enzyme activity showed that Ca+2 ions increased pullulan activity, whereas the other ions and reagents inhibited pullulanase activity. The ability of B. cereus SDK2 to produce high levels of neopullulanase stable at 60 oC that can generate panose from pullulan, make this newly isolated strain a valuable source of debranching enzyme for biotechnology, the starch bioprocess and medical industries.

  5. Isolation of Nanjing Local Strains of HHV-7 and Their Biological and Immunological Characteristics

    Institute of Scientific and Technical Information of China (English)

    KunYao; JieDou; JianXu; HuaSun; GuangyongPeng; QiangRen; XiaohuiJi

    2004-01-01

    To investigate the biological and immunological characteristics of the Nanjing local strains of HHV7, four strains of herpesvirus were isolated from saliva specimens of one healthy individual and three children suffering from a kidney disease in Nanjing. The viruses were identified by transmission electron microscopy(EM), indirect immunofluorescence assay (IFA) with a specific monoclonal antibody; nested polymerase chain reaction, restriction mapping and DNA sequencing. The virus-infected cells showed the typical cytophathic effect (CPE) under microscopy and could be detected by IFA with the human herpesvirus-7 (HHV-7) specificantibody. Under EM, herpesvirus-like and virions capsids could be found in their cytoplasm or nucleoplasm.HHV-7 DNA fragments amplified from infected cells by nested PCR were confirmed by restriction mappingand DNA sequencing. Similarly to DC strain, an known HHV-7 strain used in the present study as the positivecontrol, the virus could be inactivated by ultraviolet irradiation for 10 min, heated at 45~C for 30 min, pH 9 at 4~C for 2 h and ether or chloroform for 10 h. The virus induced the production of TNF-α, IL-10 andIL-12p70 while inhibited IFN-T secretion, increased the percentage of CD2+ cells while decreased that of CD4+or CD45RA+ cells. The results indicate that the viruses isolated in Nanjing are HHV-7, which has similarbiological characteristic to the known HHV-7 strain, DC. Infection with HHV-7 in vitro could affect immunefunction of lymphocytes by disturbing cytokine production and CD antigen expression. Cellular & MolecularImmunology. 2004;1(5):367-372.

  6. Isolation of Nanjing Local Strains of HHV-7 and Their Biological and Immunological Characteristics

    Institute of Scientific and Technical Information of China (English)

    Kun Yao; Jie Dou; Jian Xu; Hua Sun; Guangyong Peng; Qiang Ren; Xiaohui Ji

    2004-01-01

    To investigate the biological and immunological characteristics of the Nanjing local strains of HHV7, four strains of herpesvirus were isolated from saliva specimens of one healthy individual and three children suffering from a kidney disease in Nanjing. The viruses were identified by transmission electron microscopy(EM), indirect immunofluorescence assay (IFA) with a specific monoclonal antibody; nested polymerase chain reaction, restriction mapping and DNA sequencing. The virus-infected cells showed the typical cytophathic effect (CPE) under microscopy and could be detected by IFA with the human herpesvirus-7 (HHV-7) specific antibody. Under EM, herpesvirus-like and virions capsids could be found in their cytoplasm or nucleoplasm.HHV-7 DNA fragments amplified from infected cells by nested PCR were confirmed by restriction mapping and DNA sequencing. Similarly to DC strain, an known HHV-7 strain used in the present study as the positive control, the virus could be inactivated by ultraviolet irradiation for 10 min, heated at 45℃ for 30 min, pH<5 or>9 at 4℃ for 2 h and ether or chloroform for 10 h. The virus induced the production of TNF-α, IL-10 and IL-12p70 while inhibited IFN-γ secretion, increased the percentage of CD2+ cells while decreased that of CD4+ or CD45RA+ cells. The results indicate that the viruses isolated in Nanjing are HHV-7, which has similar biological characteristic to the known HHV-7 strain, DC. Infection with HHV-7 in vitro could affect immune function of lymphocytes by disturbing cytokine production and CD antigen expression. Cellular & Molecular Immunology. 2004; 1(5):367-372.

  7. Different Candida parapsilosis clinical isolates and lipase deficient strain trigger an altered cellular immune response

    Directory of Open Access Journals (Sweden)

    Renata eToth

    2015-10-01

    Full Text Available Numerous human diseases can be associated with fungal infections either as potential causative agents or as a result of changed immune status due to a primary disease. Fungal infections caused by Candida species can vary from mild to severe dependent upon the site of infection, length of exposure and past medical history. Patients with impaired immune status are at increased risk for chronic fungal infections. Recent epidemiologic studies have revealed the increasing incidence of candidiasis caused by non-albicans species such as C. parapsilosis. Due to its increasing relevance we chose two distinct C. parapsilosis strains, to describe the cellular innate immune response towards this species. In the first section of our study we compared the interaction of CLIB 214 and GA1 cells with murine and human macrophages. Both strains are commonly used to investigate C. parapsilosis virulence properties. CLIB 214 is a rapidly pseudohyphae-forming strain and GA1 is an isolate that mainly exists in a yeast form. Our results showed, that the phagocyte response was similar in terms of overall uptake, however differences were observed in macrophage migration and engulfment of fungal cells. As C. parapsilosis releases extracellular lipases in order to promote host invasion we further investigated the role of these secreted components during the distinct stages of the phagocytic process. Using a secreted lipase deficient mutant strain and the parental strain GA1 individually and simultaneously, we confirmed that fungal secreted lipases influence the fungi’s virulence by detecting altered innate cellular responses.In this study we report that two isolates of a single species can trigger markedly distinct host responses and that lipase secretion plays a role on the cellular level of host pathogen interactions.

  8. Is there ethological isolation among Drosophila melanogaster strains reared for more than 35 generations on different food?

    Directory of Open Access Journals (Sweden)

    Pavković-Lučić Sofija

    2009-01-01

    Full Text Available Ethological isolation among three strains of D. melanogaster reared for more than 35 generations on different food (standard cornmeal-agar-yeast medium, banana, and tomato was tested in a series of multiple choice tests. Long-term maintenance on different substrates did not contribute to sexual isolation among three 'nutritional strains', as degrees of ethological isolation (Malogolowkin-Cohen's isolation indices were not statistically significant. Males and females reared on different nutritional resources were equally successful in matings, i.e., significant differences in the number of copulations achieved by males and females from various lines were not observed.

  9. The inhibition of Clostridium botulinum type C by other bacteria in wetland sediments

    Science.gov (United States)

    Sandler, Renee J.; Rocke, Tonie E.; Yuill, Thomas M.

    1998-01-01

    Bacteria with inhibitory activity against Clostridium botulinum type C were isolated from 32% of sediment samples (n = 1600) collected from 10 marshes in a northern California wetland over a 12 mo period. Aerobic and anaerobic bacteria with inhibitory activity were isolated from 12% and 23% of the samples, respectively. Bacteria with inhibitory activity were isolated from all 10 study sites and throughout the year. This study demonstrates that bacteria with inhibitory activity against C. botulinum type C occur naturally in wetland sediments.

  10. Evaluation of normalization reference genes for RT-qPCR analysis of spo0A and four sporulation sigma factor genes in Clostridium botulinum Group I strain ATCC 3502.

    Science.gov (United States)

    Kirk, David G; Palonen, Eveliina; Korkeala, Hannu; Lindström, Miia

    2014-04-01

    Heat-resistant spores of Clostridium botulinum can withstand the pasteurization processes in modern food processing. This poses a risk to food safety as spores may germinate into botulinum neurotoxin-producing vegetative cells. Sporulation in Bacillus subtilis, the model organism for sporulation, is regulated by the transcription factor Spo0A and four alternative sigma factors, SigF, SigE, SigG, and SigK. While the corresponding regulators are found in available genomes of C. botulinum, little is known about their expression. To accurately measure the expression of these genes using quantitative reverse-transcriptase PCR (RT-qPCR) during the exponential and stationary growth phases, a suitable normalization reference gene is required. 16S rrn, adK, alaS, era, gluD, gyrA, rpoC, and rpsJ were selected as the candidate reference genes. The most stable candidate reference gene was 16S ribosomal RNA gene (rrn), based on its low coefficient of variation (1.81%) measured during the 18-h study time. Using 16S rrn as the normalization reference gene, the relative expression levels of spo0A, sigF, sigE, sigG, and sigK were measured over 18h. The pattern of expression showed spo0A expression during the logarithmic growth phase, followed by a drop in expression upon entry to the stationary phase. Expression levels of sigF, sigE, and sigG peaked simultaneously at the end of the exponential growth phase. Peak expression of sigK occurred at 18h, however low levels of expression were detected during the exponential phase. These findings suggest these sigma factors play a role in C. botulinum sporulation that is similar, but not equal, to their role in the B. subtilis model.

  11. Isolation and Characterization of Lytic Properties of Bacteriophages Specific for M. haemolytica Strains.

    Directory of Open Access Journals (Sweden)

    Renata Urban-Chmiel

    Full Text Available The objective of this study was isolation and morphological characterization of temperate bacteriophages obtained from M. haemolytica strains and evaluation of their lytic properties in vitro against M. haemolytica isolated from the respiratory tract of calves.The material for the study consisted of the reference strain M. haemolytica serotype 1 (ATCC® BAA-410™, reference serotypes A1, A2, A5, A6, A7, A9 and A11, and wild-type isolates of M. haemolytica. Bacteriophages were induced from an overnight bacterial starter culture of all examined M. haemolytica strains treated with mitomycin C. The lytic properties and host ranges were determined by plaque assays. The morphology of the bacteriophages was examined in negative-stained smears with 5% uranyl acetate solution using a transmission electron microscope. The genetic analysis of the bacteriophages was followed by restriction analysis of bacteriophage DNA. This was followed by analysis of genetic material by polymerase chain reaction (PCR.Eight bacteriophages were obtained, like typical of the families Myoviridae, Siphoviridae and Podoviridae. Most of the bacteriophages exhibited lytic properties against the M. haemolytica strains. Restriction analysis revealed similarities to the P2-like phage obtained from the strain M. haemolytica BAA-410. The most similar profiles were observed in the case of bacteriophages φA1 and φA5. All of the bacteriophages obtained were characterized by the presence of additional fragments in the restriction profiles with respect to the P2-like reference phage. In the analysis of PCR products for the P2-like reference phage phi-MhaA1-PHL101 (DQ426904 and the phages of the M. haemolytica serotypes, a 734-bp phage PCR product was obtained. The primers were programmed in Primer-Blast software using the structure of the sequence DQ426904 of reference phage PHL101.The results obtained indicate the need for further research aimed at isolating and characterizing

  12. Molecular characterization of invasive Neisseria meningitidis strains isolated in Chile during 2010-2011.

    Directory of Open Access Journals (Sweden)

    Gisselle N Barra

    Full Text Available BACKGROUND: With the upcoming licensure of Outer Membrane Protein-based vaccines against meningococcal disease, data on disease incidence and molecular characteristic of circulating N. meningitidis strains in Latin American countries is needed. Chile is, to date, one of the few countries in the region that has performed this type of work in a comprehensive collection of disease-associated strains from two consecutive years, 2010-2011. METHODS: A total of 119 N. meningitidis strains isolated from patients with invasive disease in Chile in 2010-2011 were characterized by the National Reference Laboratory. Serogroup determination, MLST and porA typing were performed. RESULTS: Serogroup B was predominant in both study years, but W135 experienced a noticeable increase in 2011 compared to 2010. ST-11 complex, ST-41/44 complex ST-32 complex were the most prevalent among the isolates, and were strongly associated with serogroups W135 (ST-11 Complex and B (ST-41/44 and ST-32 complexes. Likewise, the major porA types detected were strongly associated with these three clonal complexes: P1.5,2 was found exclusively among W135:ST-11 isolates, whereas P1.7, 2-3 was only detected in C:ST-11. ST-41/44 isolates mainly had P1.10-8, and ST-32 complex were associated with a P1.18-8 porA. CONCLUSIONS: Our data show disease-associated N. meningitidis circulating in Chile are similar to those found in other parts of the world. The increase on W135:ST-11 isolates observed in 2011 foretold the unusual epidemiological situation experienced in the country in 2012, and MLST data show that this strain is indistinguishable from the one linked to the global Hajj 2000-related outbreak that occurred in 2001. Finally, this work demonstrates the importance of maintaining a strong national surveillance program integrating clinical, epidemiological and laboratory data and incorporating gold standard diagnostic and characterization techniques that allow the data to be compared all

  13. Evaluating synergy between marbofloxacin and gentamicin in Pseudomonas aeruginosa strains isolated from dogs with otitis externa.

    Science.gov (United States)

    Jerzsele, Ákos; Pásztiné-Gere, Erzsébet

    2015-03-01

    The aim of this study was to determine antimicrobial susceptibility of Pseudomonas aeruginosa strains to marbofloxacin and gentamicin, and investigate the possible synergistic, additive, indifferent or antagonistic effects between the two agents. P. aeruginosa strains can develop resistance quickly against certain antibiotics if used alone, thus the need emerges to find synergistic combinations. A total of 68 P. aeruginosa strains isolated from dogs were examined. In order to describe interactions between marbofloxacin and gentamicin the checkerboard microdilution method was utilized. The MICs (minimum inhibitory concentrations) for marbofloxacin and gentamicin were in the range 0.25-64 mg/L and 0.25-32 mg/L, respectively. The combination of marbofloxacin and gentamicin was more effective with a MIC range of 0.031-8 mg/L and a MIC90 of 1 mg/L, compared to 16 mg/L for marbofloxacin alone and 8 mg/L for gentamicin alone. The FIC (fractional inhibitory concentration) indices ranged from 0.0945 (pronounced synergy) to 1.0625 (indifference). Synergy between marbofloxacin and gentamicin was found in 33 isolates. The mean FIC index is 0.546, which represents a partial synergistic/additive effect close to the full synergy threshold. In vitro results indicate that marbofloxacin and gentamicin as partially synergistic agents may prove clinically useful in combination therapy against P. aeruginosa infections. Although marbofloxacin is not used in the human practice, the interactions between fluoroquinolones and aminoglycosides may have importance outside the veterinary field.

  14. Antimicrobial Resistant Pattern of Escherichia Coli Strains Isolated from Pediatric Patients in Jordan

    Directory of Open Access Journals (Sweden)

    Mohammad Alshara

    2011-05-01

    Full Text Available The present study was conducted to investigate antimicrobial resistant pattern of Escherichia coli (E. coli strains isolated from clinical specimens of Jordanian pediatric patients during the period from January to December 2008. A total of 444 E. coli strains were isolated from clinical specimens and tested for their susceptibility to different antimicrobial drugs. Overall, high resistance rate was observed for ampicillin (84%, followed by amoxicillin-clavulanic acid (74.3%, cotrimoxazole (71%, nalidixic acid (47.3%, cephalothin (41%. Lower resistance rates were observed for amikacin (0% followed by Cefotaxime (11%, Ceftriaxone (11.7%, ciprofloxacin (14.5%, Norfloxacin (16.5%, gentamicin (17.3% cephalexin (20.9%, Ceftazidime (22.5%, cefixime (29.6%, and cefaclor (32.8%. Ampicillin, amoxicillin-clavulanic acid and cotrimoxazole were found to be ineffective at in vitro inhibition of the E. coli of pediatric origin. Amikacin was highly effective for E. coli with susceptibility rate of 100%. The majority of E. coli strains were susceptible to third generation cephalosporins and fluoroquinolones.

  15. Phenotype and genotype of Candida albicans strains isolated from pregnant women with recurrent vaginitis.

    Science.gov (United States)

    Maffei, C M; Paula, C R; Mazzocato, T S; Franceschini, S

    1997-01-01

    Fourteen out of 80 pregnant women receiving prenatal care presented signs and symptoms of recurrent vaginal candidiasis. Candida albicans strains were isolated from 12 patients (85.7%), and these were submitted to morphotyping (morphological characteristics of the colony), antifungal typing (pattern of sensitivity to amphotericin B, 5-fluorcytosine, myconazole, ketoconazole and fluconazole) and genotyping (electrophoretic migration of DNA fragments digested with EcoRI and HinfI). Alteration of morphotype and antifungal type was observed in 50% of the patients, but the genotype of the strains isolated from the same patients at different times was identical in all subjects. The predominant morphotypes presented continuous fringes and the basic changes observed among antifungal types was the emergence of strains resistant to myconazole, which was the drug used for the treatment of the first episode of vaginitis. We conclude that recurrent vaginal candidasis is caused by the persistence of a single yeast genotype that undergoes morphological and behavioral changes in the presence of antifungal agents due to the selective pressure to which it is submitted.

  16. Isolation of a yeast strain able to produce a polygalacturonase with maceration activity of cassava roots

    Directory of Open Access Journals (Sweden)

    María Alicia Martos

    2013-06-01

    Full Text Available The objective of the present study was the isolation of a yeast strain, from citrus fruit peels, able to produce a polygalacturonase by submerged fermentation with maceration activity of raw cassava roots. Among 160 yeast strains isolated from citrus peels, one strain exhibited the strongest pectinolytic activity. This yeast was identified as Wickerhamomyces anomalus by 5.8S-ITS RFLP analysis and confirmed by amplification of the nucleotide sequence. The yeast produced a polygalacturonase (PG in Erlenmeyer shake flasks containing YNB, glucose, and citrus pectin. PG synthesis occurred during exponential growth phase, reaching 51 UE.mL-1 after 8 hours of fermentation. A growth yield (Yx/s of 0.43 gram of cell dry weight per gram of glucose consumed was obtained, and a maximal specific growth rate (µm of 0.346 h-1 was calculated. The microorganism was unable to assimilate sucrose, galacturonic acid, polygalacturonic acid, or citrus pectin, but it required glucose as carbon and energy source and polygalacturonic acid or citrus pectin as inducers of enzyme synthesis. The crude enzymatic extract of Wickerhamomyces anomalus showed macerating activity of raw cassava. This property is very important in the production of dehydrated mashed cassava, a product of regional interest in the province of Misiones, Argentina.

  17. Photodynamic inactivation of virulence factors of Candida strains isolated from patients with denture stomatitis.

    Science.gov (United States)

    Pereira, Cristiane Aparecida; Domingues, Nádia; Silva, Michelle Peneluppi; Costa, Anna Carolina Borges Pereira; Junqueira, Juliana Campos; Jorge, Antonio Olavo Cardoso

    2015-12-01

    Candida species are major microorganisms isolated in denture stomatitis (DS), an inflammatory process of the mucosa underlying removable dental prostheses, and express a variety of virulence factors that can increase their pathogenicity. The potential of Photodynamic inactivation (PDI) in planktonic culture, biofilms and virulence factors of Candida strains was evaluated. A total of 48 clinical Candida isolates from individuals wearing removable maxillary prostheses with DS were included in the study. The effects of erythrosine (ER, 200 μM) and a green LED (λ 532 ± 10 nm, 237 mW/cm(2) and 42.63 J/cm(2)) in a planktonic culture were evaluated. The effect of the addition of ER at a concentration of 400 μM together with a green LED was evaluated in biofilms. The virulence factors of all of the Candida strains were evaluated before and after the PDI process in cells derived from biofilm and planktonic assays. All of the Candida species were susceptible to ER and green LED. However, the biofilm structures were more resistant to PDI than the planktonic cultures. PDI also promoted slight reductions in most of the virulence factors of C. albicans and some of the Candida tropicalis strains. These results suggest that the addition of PDI is effective for reducing yeasts and may also reduce the virulence of certain Candida species and decrease their pathogenicity.

  18. Modification of norfloxacin by a Microbacterium sp. strain isolated from a wastewater treatment plant.

    Science.gov (United States)

    Kim, Dae-Wi; Heinze, Thomas M; Kim, Bong-Soo; Schnackenberg, Laura K; Woodling, Kellie A; Sutherland, John B

    2011-09-01

    Antimicrobial residues found in municipal wastewater may increase selective pressure on microorganisms for development of resistance, but studies with mixed microbial cultures derived from wastewater have suggested that some bacteria are able to inactivate fluoroquinolones. Medium containing N-phenylpiperazine and inoculated with wastewater was used to enrich fluoroquinolone-modifying bacteria. One bacterial strain isolated from an enrichment culture was identified by 16S rRNA gene sequence analysis as a Microbacterium sp. similar to a plant growth-promoting bacterium, Microbacterium azadirachtae (99.70%), and a nematode pathogen, "M. nematophilum" (99.02%). During growth in medium with norfloxacin, this strain produced four metabolites, which were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) analyses as 8-hydroxynorfloxacin, 6-defluoro-6-hydroxynorfloxacin, desethylene norfloxacin, and N-acetylnorfloxacin. The production of the first three metabolites was enhanced by ascorbic acid and nitrate, but it was inhibited by phosphate, amino acids, mannitol, formate, and thiourea. In contrast, N-acetylnorfloxacin was most abundant in cultures supplemented with amino acids. This is the first report of defluorination and hydroxylation of a fluoroquinolone by an isolated bacterial strain. The results suggest that some bacteria may degrade fluoroquinolones in wastewater to metabolites with less antibacterial activity that could be subject to further degradation by other microorganisms.

  19. Modification of Norfloxacin by a Microbacterium sp. Strain Isolated from a Wastewater Treatment Plant▿

    Science.gov (United States)

    Kim, Dae-Wi; Heinze, Thomas M.; Kim, Bong-Soo; Schnackenberg, Laura K.; Woodling, Kellie A.; Sutherland, John B.

    2011-01-01

    Antimicrobial residues found in municipal wastewater may increase selective pressure on microorganisms for development of resistance, but studies with mixed microbial cultures derived from wastewater have suggested that some bacteria are able to inactivate fluoroquinolones. Medium containing N-phenylpiperazine and inoculated with wastewater was used to enrich fluoroquinolone-modifying bacteria. One bacterial strain isolated from an enrichment culture was identified by 16S rRNA gene sequence analysis as a Microbacterium sp. similar to a plant growth-promoting bacterium, Microbacterium azadirachtae (99.70%), and a nematode pathogen, “M. nematophilum” (99.02%). During growth in medium with norfloxacin, this strain produced four metabolites, which were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR) analyses as 8-hydroxynorfloxacin, 6-defluoro-6-hydroxynorfloxacin, desethylene norfloxacin, and N-acetylnorfloxacin. The production of the first three metabolites was enhanced by ascorbic acid and nitrate, but it was inhibited by phosphate, amino acids, mannitol, formate, and thiourea. In contrast, N-acetylnorfloxacin was most abundant in cultures supplemented with amino acids. This is the first report of defluorination and hydroxylation of a fluoroquinolone by an isolated bacterial strain. The results suggest that some bacteria may degrade fluoroquinolones in wastewater to metabolites with less antibacterial activity that could be subject to further degradation by other microorganisms. PMID:21724893

  20. Screening of Molecular Virulence Markers in Staphylococcus aureus and Pseudomonas aeruginosa Strains Isolated from Clinical Infections

    Science.gov (United States)

    Cotar, Ani-Ioana; Chifiriuc, Mariana-Carmen; Dinu, Sorin; Bucur, Marcela; Iordache, Carmen; Banu, Otilia; Dracea, Olguta; Larion, Cristina; Lazar, Veronica

    2010-01-01

    Staphylococcus (S.) aureus and Pseudomonas (Ps.) aeruginosa are two of the most frequently opportunistic pathogens isolated in nosocomial infections, responsible for severe infections in immunocompromised hosts. The frequent emergence of antibiotic-resistant S. aureus and Ps. aeruginosa strains has determined the development of new strategies in order to elucidate the different mechanisms used by these bacteria at different stages of the infectious process, providing the scientists with new procedures for preventing, or at least improving, the control of S. aureus and Ps. aeruginosa infections. The purpose of this study was to characterize the molecular markers of virulence in S. aureus and Ps. aeruginosa strains isolated from different clinical specimens. We used multiplex and uniplex PCR assays to detect the genes encoding different cell-wall associated and extracellular virulence factors, in order to evaluate potential associations between the presence of putative virulence genes and the outcome of infections caused by these bacteria. Our results demonstrate that all the studied S. aureus and Ps. aeruginosa strains synthesize the majority of the investigated virulence determinants, probably responsible for different types of infections. PMID:21614207

  1. Screening of Molecular Virulence Markers in Staphylococcus aureus and Pseudomonas aeruginosa Strains Isolated from Clinical Infections

    Directory of Open Access Journals (Sweden)

    Veronica Lazar

    2010-12-01

    Full Text Available Staphylococcus (S. aureus and Pseudomonas (Ps. aeruginosa are two of the most frequently opportunistic pathogens isolated in nosocomial infections, responsible for severe infections in immunocompromised hosts. The frequent emergence of antibiotic-resistant S. aureus and Ps. aeruginosa strains has determined the development of new strategies in order to elucidate the different mechanisms used by these bacteria at different stages of the infectious process, providing the scientists with new procedures for preventing, or at least improving, the control of S. aureus and Ps. aeruginosa infections. The purpose of this study was to characterize the molecular markers of virulence in S. aureus and Ps. aeruginosa strains isolated from different clinical specimens. We used multiplex and uniplex PCR assays to detect the genes encoding different cell-wall associated and extracellular virulence factors, in order to evaluate potential associations between the presence of putative virulence genes and the outcome of infections caused by these bacteria. Our results demonstrate that all the studied S. aureus and Ps. aeruginosa strains synthesize the majority of the investigated virulence determinants, probably responsible for different types of infections.

  2. Mercury remediation potential of a mercury resistant strain Sphingopyxis sp. SE2 isolated from contaminated soil.

    Science.gov (United States)

    Mahbub, Khandaker Rayhan; Krishnan, Kannan; Naidu, Ravi; Megharaj, Mallavarapu

    2017-01-01

    A mercury resistant bacterial strain SE2 was isolated from contaminated soil. The 16s rRNA gene sequencing confirms the strain as Sphingopyxis belongs to the Sphingomonadaceae family of the α-Proteobacteria group. The isolate showed high resistance to mercury with estimated concentrations of Hg that caused 50% reduction in growth (EC50) of 5.97 and 6.22mg/L and minimum inhibitory concentrations (MICs) of 32.19 and 34.95mg/L in minimal and rich media, respectively. The qualitative detection of volatilized mercury and the presence of mercuric reductase enzyme proved that the strain SE2 can potentially remediate mercury. ICP-QQQ-MS analysis of the remaining mercury in experimental broths indicated that a maximum of 44% mercury was volatilized within 6hr by live SE2 culture. Furthermore a small quantity (23%) of mercury was accumulated in live cell pellets. While no volatilization was caused by dead cells, sorption of mercury was confirmed. The mercuric reductase gene merA was amplified and sequenced. Homology was observed among the amino acid sequences of mercuric reductase enzyme of different organisms from α-Proteobacteria and ascomycota groups.

  3. Phylogenetic evidence for multiple intertypic recombinations in enterovirus B81 strains isolated in Tibet, China.

    Science.gov (United States)

    Hu, Lan; Zhang, Yong; Hong, Mei; Zhu, Shuangli; Yan, Dongmei; Wang, Dongyan; Li, Xiaolei; Zhu, Zhen; Tsewang; Xu, Wenbo

    2014-08-12

    Enterovirus B81 (EV-B81) is a newly identified serotype within the species enterovirus B (EV-B). To date, only eight nucleotide sequences of EV-B81 have been published and only one full-length genome sequence (the prototype strain) has been made available in the GenBank database. Here, we report the full-length genome sequences of two EV-B81 strains isolated in the Tibet Autonomous Region of China during acute flaccid paralysis surveillance activities, and we also conducted an antibody seroprevalence study in two prefectures of Tibet. The sequence comparison and phylogenetic dendrogram analysis revealed high variability among the global EV-B81 strains and frequent intertypic recombination in the non-structural protein region of EV-B serotypes, suggesting high genetic diversity of EV-B81. However, low positive rates and low titers of neutralizing antibodies against EV-B81 were detected. Nearly 68% of children under the age of five had no neutralizing antibodies against EV-B81. Hence, the extent of transmission and the exposure of the population to this EV type are very limited. Although little is known about the biological and pathogenic properties of EV-B81 because of few research in this field owing to the limited number of isolates, our study provides basic information for further studies of EV-B81.

  4. Isolation and pathogenic analysis of virulent Marek's disease virus field strain in China.

    Science.gov (United States)

    Cui, Ning; Su, Shuai; Sun, Peng; Zhang, Yankun; Han, Ni; Cui, Zhizhong

    2016-07-01

    Marek's disease (MD) has become increasingly common in China, resulting in considerable economic loss. The etiological agent is unclear. In this study, we isolated a field MD virus (MDV) strain, designated SX1301, from CVI988/Rispens-vaccinated chickens with tumors. Co-infection of avian leukosis virus, reticuloendotheliosis virus, and chicken infectious anemia virus was excluded by polymerase chain reaction, enzyme-linked immunosorbant assay, DNA blotting hybridization, and indirect immunofluorescence assay. As with most strains isolated in China, SX1301 had the same amino acid mutation of meq protein at positions 77(E), 80(Y), and 115(A) Animal experimental results showed development of lethal MD in 57% and MD tumor in 23% of the specific pathogen-free chickens inoculated with SX1301, with tumors mainly distributed in spleen, liver, and kidney. CVI988/Rispens protected 83% of chickens upon challenge with SX1301, with a mortality rate and tumor incidence of 10% and 7%, respectively. These results implicated SX1301 as a virulent MDV strain, with commercial MDV vaccine CVI988/Rispens unable to confer adequate protection against SX1301. There have been no reports of very virulent (vv) plus MDV in China, but frequently occurring virulent MDV may account for the repeated outbreaks of MD. Vaccines with greater efficacy are needed to protect against MDV.

  5. Isolation and genotyping of acanthamoeba strains from soil sources from Jamaica, West Indies.

    Science.gov (United States)

    Todd, Cheridah D; Reyes-Batlle, María; Martín-Navarro, Carmen Ma; Dorta-Gorrín, Alexis; López-Arencibia, Atteneri; Martínez-Carretero, Enrique; Piñero, José E; Valladares, Basilio; Lindo, John F; Lorenzo-Morales, Jacob

    2015-01-01

    Acanthamoeba spp. are opportunistic pathogens that are ubiquitous in nature. Many species of this genus are responsible for a fatal encephalitis and keratitis in humans and other animals. Seventy-two soil samples were collected from the parishes across Jamaica and assessed for the presence of Acanthamoeba spp. Cultivation was carried out on non-nutrient agar plates seeded with heat killed Escherichia coli. PCR and sequencing of the DF3 region were carried out in order to genotype the isolated strains of Acanthamoeba. Thermotolerance and osmotolerance assays were utilized to investigate the pathogenic potential of the Acanthamoeba isolates. Acanthamoeba spp. was isolated from 63.9% of soil samples. Sequencing of the DF3 region of the 18S rDNA resulted in the identification of genotypes T4, T5, and T11. T4 genotype was most frequently isolated. Most isolates were thermotolerant or both thermotolerant and osmotolerant, indicating that they may present the potential to cause disease in humans and other animals.

  6. Multilocus sequence typing (MLST) of leptospiral strains isolated from two geographic locations of Tamil Nadu, India.

    Science.gov (United States)

    Kanagavel, Murugesan; Princy Margreat, Alphonse Asirvatham; Arunkumar, Manivel; Prabhakaran, Shanmugarajan Gnanasekaran; Shanmughapriya, Santhanam; Natarajaseenivasan, Kalimuthusamy

    2016-01-01

    Here the rodent carrier status for the transmission of human leptospirosis in Tiruchirappalli, district, Tamil Nadu, India was assessed. The predominantly circulating leptospiral STs were recognized by multilocus sequence typing (MLST). A total of 113 rodents were trapped from different provinces of the Tiruchirappalli district. The most prevalent rodent was Bandicota bengalensis (37.2%), and of the total, 52.2% (n=59) rodents were found to be positive for leptospiral 16S rRNA. These results were validated with a leptospiral culture positivity of 45.8% (n=27). Three isolates from Chennai (2 rodents and 1 human) and 1 human isolate from Tiruchirappalli were included to understand the spatial variations and to track the source of human leptospirosis. The serogroup, serovar, and species level identification of all 31 isolates identified 28 to be Leptospira borgpetersenii serovar Javanica and three as Leptospira interrogans serovar Autumnalis. MLST analysis defined all isolates to the existing ST profiles (ST145 and ST27) with the exception of 6 L. borgpetersenii (ST DR) isolates that showed variations in the sucA and pfkB loci. The DR ST was locally confined to Chatram province of Tiruchirappalli suggesting an epidemiological link. The predominant STs, ST145 and ST-DR form a group, indicating the presence of original strain that subsequently diverged evolutionarily into two STs. The variations between L. borgpetersenii in sucA and pfkB loci may be an indication that evolutionary changes transpired in Tiruchirappalli.

  7. Complete Genome Sequence of Xanthomonas arboricola pv. juglandis 417, a Copper-Resistant Strain Isolated from Juglans regia L.

    Science.gov (United States)

    Pereira, Ulisses P; Gouran, Hossein; Nascimento, Rafael; Adaskaveg, James E; Goulart, Luiz Ricardo; Dandekar, Abhaya M

    2015-01-01

    Here, we report the complete genome sequence of Xanthomonas arboricola pv. juglandis 417, a copper-resistant strain isolated from a blighted walnut fruit (Juglans regia L. cv. Chandler). The genome consists of a single chromosome (5,218 kb).

  8. Draft Genome Sequence of Clostridium bifermentans Strain WYM, a Promising Biohydrogen Producer Isolated from Landfill Leachate Sludge.

    Science.gov (United States)

    Wong, Y M; Juan, J C; Gan, H M; Austin, C M

    2014-03-06

    Clostridium bifermentans strain WYM is an effective biohydrogen producer isolated from landfill leachate sludge. Here, we present the assembly and annotation of its genome, which may provide further insights into the metabolic pathways involved in efficient biohydrogen production.

  9. Genetic Analysis of the VP1 Region of Human Enterovirus 71 Strains Isolated in Fuyang, China, During 2008

    Institute of Scientific and Technical Information of China (English)

    Shao-hui MA; Jian-sheng LIU; Jing-jing WANG; Hai-jing SHI; Hui-juan YANG; Jun-ying CHEN; Long-ding LIU; Qi-han LI

    2009-01-01

    Enterovirus 71 (EV71) is a common cause of Hand, foot, and mouth disease (HFMD) and may also cause severe neurological diseases, such as encephalitis and poliomyelitis-like paralysis. To examine the genetic diversity of EV71, we determined and analyzed the complete VP1 sequences (891 nucleotides) from nine EV71 strains isolated in Fuyang, China. We found that nine EV71 strains isolated were over 98% homologous at the nucleotide level and 93%-100% homologous to members of the C4 subgenogroup. At the amino acid level, these Fuyang strains were 99% -100% homologous to one another, 97%-100% homologous to members of the C4 subgenogroup, and the histidine(H) at amino acid position 22 was conserved among the Fuyang strains. The results indicate that Fuyang isolates belong to genotype C4, and an H at position 22 appears to be a marker for the Fuyang strains.

  10. Molecular typing of Legionella pneumophila serogroup 1 clinical strains isolated in Italy.

    Science.gov (United States)

    Fontana, Stefano; Scaturro, Maria; Rota, Maria Cristina; Caporali, Maria Grazia; Ricci, Maria Luisa

    2014-07-01

    Molecular typing methods for discriminating different bacterial isolates are essential epidemiological tools in prevention and control of Legionella infections and outbreaks. A selection of 56 out of 184 Legionella pneumophila serogroup 1 (Lp1) clinical isolates, collected from different Italian regions between 1987 and 2012, and stored at the National Reference Laboratory for Legionella, were typed by monoclonal antibody (MAb) subgrouping, amplified fragment length polymorphism (AFLP) and sequence based typing (SBT). These strains were isolated from 39 community (69.6%), 14 nosocomial (25%) and 3 travel associated (5.4%) Legionnaires'disease cases. MAb typing results showed a prevalence of MAb 3/1 positive isolates (75%) with the Philadelphia subgroup representing 35.7%, followed by Knoxville (23.2%), Benidorm (12.5%), Allentown/France (1.8%), Allentown/France-Philadelphia (1.8%). The remaining 25% were MAb 3/1 negative, namely 11 Olda (19.6%), 2 Oxford (3.6%) and 1 Bellingham (1.8%) subgroups. AFLP analysis detected 20 different genomic profiles. SBT analysis revealed 32 different sequence types (STs) with high diversity of STs (IODSTs=0.952) 12 of which were never described before. ST1 and ST23 were most frequently isolated as observed worldwide. A helpful analysis of data from SBT, MAb subgrouping and AFLP is provided, as well as a comparison to the Lp1 types investigated from other countries. This study describes the first Italian Lp1 strains database, providing molecular epidemiology data useful for future epidemiological investigations, especially of travel associated Legionnaires' diseases (TALD) cases, Italy being the country associated with the highest number of clusters.

  11. Continuous gluconic acid production by isolated yeast-like mould strains of Aureobasidium pullulans.

    Science.gov (United States)

    Anastassiadis, S; Aivasidis, A; Wandrey, C

    2003-04-01

    By extensive microbial screening, about 50 strains with the ability to secrete gluconic acid were isolated from wild flowers. The strains belong to the yeast-like mould Aureobasidium pullulans (de Bary) Arnaud. In shake flask experiments, gluconic acid concentrations between 23 and 140 g/l were produced within 2 days using a mineral medium. In batch experiments, various important fermentation parameters influencing gluconic acid production by A. pullulans isolate 70 (DSM 7085) were identified. Continuous production of gluconic acid with free-growing cells of the isolated yeast-like microorganisms was studied. About 260 g/l gluconic acid at total glucose conversion could be achieved using continuous stirred tank reactors in defined media with residence times (RT) of about 26 h. The highest space-time-yield of 19.3 g l(-1) x h(-1)) with a gluconic acid concentration of 207.5 g/l was achieved with a RT of 10.8 h. The possibility of gluconic acid production with biomass retention by immobilised cells on porous sinter glass is discussed. The new continuous gluconate fermentation process provides significant advantages over traditional discontinuous operation employing Aspergillus niger. The aim of this work was the development of a continuous fermentation process for the production of gluconic acid. Process control becomes easier, offering constant product quality and quantity.

  12. Antagonist capacity of Newly Isolated Strains of Pseudomonas Fluorescens against Three Important Phytopathogenic Bacteria

    Directory of Open Access Journals (Sweden)

    Reynaldo D.L. Cruz-Quiroz

    2011-01-01

    Full Text Available Problem statement: Phytopatogenic bacteria cause several damages to plants with important economical consequences. They provoke losses of product quality affecting all commercial chain of crops, for this reason, their control is a priority. Approach: We evaluated antagonist capacity of newly isolated Pseudonomas fluorescens strains against three important phytopatogenic bacteria (Clavibacter michiganensis, Xanthomonas axonopodis and Erwinia carotovora. Soils from commercial cropping of Capsicum annum L of several Mexican regions were used to isolate P. fluorescens strains. Results: Isolates producing flourescein were purified on King B agar and biochemically identified. Crude extracts with and without cells were produced in King B broths and their antagonist capacities were evaluated by the plate diffusion procedure on nutritive agar. Conclusion: Obtained results demonstrated that cell free extracts exhibited a limited antagonist capacity in comparison of those extracts with cells, which showed an excellent capacity to inhibit the growth of C. michiganensis, X. axonopodis and E. carotovora, demonstrating the intracellular nature of the bioactive metabolites associated to bacterial growth inhibition.

  13. Serotype distribution, antibiotic susceptibility, and genetic relatedness of Neisseria meningitidis strains recently isolated in Italy.

    Science.gov (United States)

    Mastrantonio, Paola; Stefanelli, Paola; Fazio, Cecilia; Sofia, Tonino; Neri, Arianna; La Rosa, Giuseppina; Marianelli, Cinzia; Muscillo, Michele; Caporali, Maria Grazia; Salmaso, Stefania

    2003-02-15

    The availability of new polysaccharide-protein conjugate vaccines against Neisseria meningitidis serogroup C prompted European National Health authorities to carefully monitor isolate characteristics. In Italy, during 1999-2001, the average incidence was 0.4 cases per 100,000 inhabitants. Serogroup B was predominant and accounted for 75% of the isolates, followed by serogroup C with 24%. Serogroup C was isolated almost twice as frequently in cases of septicemia than in cases of meningitis, and the most common phenotypes were C:2a:P1.5 and C:2b:P1.5. Among serogroup B meningococci, the trend of predominant phenotypes has changed from year to year, with a recent increase in the frequency of B:15:P1.4. Only a few meningococci had decreased susceptibility to penicillin, and, in the penA gene, all of these strains had exogenous DNA blocks deriving from the DNA of commensal Neisseria flavescens, Neisseria cinerea, and Neisseria perflava/sicca. Fluorescent amplified fragment-length polymorphism analysis revealed the nonclonal nature of the strains with decreased susceptibility to penicillin.

  14. Identification of yeast strains isolated from marcha in Sikkim, a microbial starter for amylolytic fermentation.

    Science.gov (United States)

    Tsuyoshi, Naoko; Fudou, Ryosuke; Yamanaka, Shigeru; Kozaki, Michio; Tamang, Namrata; Thapa, Saroj; Tamang, Jyoti P

    2005-03-15

    Marcha or murcha is a traditional amylolytic starter used to produce sweet-sour alcoholic drinks, commonly called jaanr in the Himalayan regions of India, Nepal, Bhutan, and Tibet (China). The aim of this study was to examine the microflora of marcha collected from Sikkim in India, focusing on yeast flora and their roles. Twenty yeast strains were isolated from six samples of marcha and identified by genetic and phenotypic methods. They were first classified into four groups (Group I, II, III, and IV) based on physiological features using an API test. Phylogenetic, morphological, and physiological characterization identified the isolates as Saccharomyces bayanus (Group I); Candida glabrata (Group II); Pichia anomala (Group III); and Saccharomycopsis fibuligera, Saccharomycopsis capsularis, and Pichia burtonii (Group IV). Among them, the Group I, II, and III strains produced ethanol. The isolates of Group IV had high amylolytic activity. Because all marcha samples tested contained both starch degraders and ethanol producers, it was hypothesized that all four groups of yeast (Group I, II, III, and IV) contribute to starch-based alcohol fermentation.

  15. Molecular Typing of Pseudomonas aeruginosa Strains Isolated from Burn Patients in South of Iran

    Directory of Open Access Journals (Sweden)

    Aziz Japoni

    2016-01-01

    Full Text Available Background: Pseudomonas aeruginosa is one of the main etiological agents in burn infections which could be life threatening for the infected patients. The aim of the present study was to identify and track source of infections using two molecular typing methods. Materials and Methods: Seventy-four strains of P. aeruginosa were isolated from burn patients and hospital environment in Ghotbadden Burn Hospital, Shiraz, Iran. Isolates were typed by arbitrary primed-polymerase chain reaction (AP-PCR and plasmid profiling. Similarity and clustering of the strains was assessed using NTSYS-PC software and photo Capt Mw program. Results: Thirty eight plasmid profiles were obtained and classified them into: 2, 3and 5 clusters, based on 50%, 64.7% and 67.5% similarity on the plotted dendrogram, respectively. Drawn dendrogarm categorized AP-PCR products to 47 different types. Conclusion: Based on these results, a limited number of P. aeruginosa types are predominant in the hospitals which infect the burn patients. To control of the infections in patients with antibiotics, resistant isolates, strong disinfection of patients’ bathroom after scrubbing of patients wounds, should be implemented.

  16. Nitrosomonas communis strain YNSRA, an ammonia-oxidizing bacterium, isolated from the reed rhizoplane in an aquaponics plant.

    Science.gov (United States)

    Tokuyama, Tatsuaki; Mine, Atsusi; Kamiyama, Kaoru; Yabe, Ryuichi; Satoh, Kazuo; Matsumoto, Hirotoshi; Takahashi, Reiji; Itonaga, Koji

    2004-01-01

    An ammonia-oxidizing bacterium (strain YNSRA) was isolated from the rhizoplane of the reed (Phragmites communis) used in an aquaponics plant which is a wastewater treatment plant. Strain YNSRA was identified as Nitrosomonas communis by taxonomic studies. The hydroxylamine-cytochrome c reductase (HCR) of strain YNSRA was found to have a higher activity (25.60 u/mg) than that of Nitrosomonas europaea ATCC25978T (8.94 u/mg). Ribulose-1,5-bisphosphate carboxylase (RubisCO) activity was detected at very low levels in strain YNSRA, whereas strain ATCC25978T had definite activity.

  17. Seroprevalence, Isolation, Genotyping, and Pathogenicity of Toxoplasma gondii Strains from Sheep in China

    Science.gov (United States)

    Yang, YuRong; Feng, YongJie; Yao, QiuXia; Wang, YingHua; Lu, YaoYao; Liang, HongDe; Zhu, XingQuan; Zhang, LongXian

    2017-01-01

    Toxoplasma gondii is an important cause of reproductive failure in small ruminants that also poses a risk to consumers who consume undercooked meat. However, little is known about sheep toxoplasmosis in China for the world. Therefore, this study was conducted to assess the prevalence of T. gondii infection in sheep from China, to isolate T. gondii via bioassay in mice and to evaluate the virulence of the isolated T. gondii based on vero cell invasion and mice. A total of 840 samples (304 unfrozen hearts and 536 sera) from sheep in China were collected from 2014 to 2016. Heart samples (n = 36) of T. gondii seropositive sheep (MAT, ≥25) were bioassayed in mice individually. DNA derived from cell cultured tachyzoites of the isolated T. gondii was characterized by PCR-RFLP of 10 loci (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). The virulence of the T. gondii was evaluated based on the mortality and encystation in mice, as well as their growth characteristics in cell culture. Antibodies to T. gondii were found in 174 of 840 (20.71%, 304 hearts juice and 536 sera) sheep by the modified agglutination test (cut-off 1:25). Viable T. gondii was isolated from the hearts of two of 36 seropositive sheep hearts. Both genotypes of the sheep heart isolates were ToxoDB#9. The virulence of the two ToxoDB#9 isolations varied significantly. To the best of our knowledge, this is the first report of isolation of ToxoDB#9 strain of T. gondii from sheep in China. PMID:28217119

  18. Antibacterial, Antifungal and Anticancer Activity of Five Strains of Soil Microorganisms Isolated From Tangkuban Perahu Mountain by Fermentation

    OpenAIRE

    Desak Gede Sri Andayani; Ukan Sukandar; Elin Y. Sukandar; I Ketut Adnyana

    2015-01-01

    Microorganisms were isolated from soil taken from Tangkuban Perahu mountain. Five strains were investigated in this study, designated TP1, TP2, TP3, TP4, and TP5, respectively. Morphological, biochemical and molecular identifications were conducted for all five strains. These isolates were shown to be closely related to Nocardia sp. YIM 65630 (90%), Streptomyces galbus (99%), Aspergillus unguis (86%), Paecilomyces marquandii (100%) and Nocardia niigatensis (95%), respectively. Production of a...

  19. Antimicrobial resistance of 100 Salmonella strains isolated from Gallus gallus in 4 wilayas of Algeria.

    Science.gov (United States)

    Bounar-Kechih, S; Hamdi, T M; Mezali, L; Assaous, F; Rahal, K

    2012-05-01

    This study aims at identifying serotypes and surveying the antimicrobial resistance and plasmid support of resistance of 100 Salmonella strains, which were isolated from 96 out of 506 (18.97%) samples taken from different production farms in the wilayas (i.e., Algerian states) of Tizi-Ouzou, Bouira, Bejaïa, and Boumerdes in 2007. The highest percentage of Salmonella (48%) was recorded in Bouira. Thirteen serotypes were identified among the 100 Salmonella strains used in this study. The most prevalent ones were Salmonella Heidelberg (24%), Salmonella Enteritidis (20%), Salmonella Albany (16%), and Salmonella Typhimurium (9%). The strains showed resistance to 8 of the 34 antibiotics tested. Fifty-three percent of strains were resistant to at least one antibiotic, among which 15.09% were multiresistant. The most frequently observed resistance was to quinolones (58.49%), with a contribution of 94.74% of Salmonella Heidelberg resistant strains. The plasmid transfer performed on 53 strains showed that only 11 exhibited one or more markers of resistance, the most frequent being ampicillin, followed by tetracycline, then cotrimoxazole, sulphonamides, and kanamycin, in that order. The tetracycline characteristics were present in 72.72% of transconjugants, those of the β-lactams and sulphonamides in 27.27% each and those of the aminosides in 9.09%. The incompatibility groups of plasmids belong to the F1me and Com1 classes, and the molecular weight of the plasmid DNA was greater than 100 kb. The phenotypic and genotypic results indicate a clonal dissemination in the Gallus gallus species in this particular study; this phenomenon could generate resistant bacteria and transferable genes of resistance to humans.

  20. Shigella sonnei strains isolated from U.S. summer students in Guadalajara, Mexico, from 1986 to 1992.

    Science.gov (United States)

    Scerpella, E G; Mathewson, J J; DuPont, H L; Marani, S K; Ericsson, C D

    1994-10-01

    Plasmid DNA analysis and antibiotic susceptibilities were used to study strains of Shigella sonnei isolated from U.S. travelers to Guadalajara, Mexico, over a period of seven years (1986 to 1992). One hundred sixty-one isolates were analyzed. By the use of cluster analysis, eight different plasmid profiles were identified during this interval. At any point in time, three to seven different plasmid profiles were present in this population. The introduction of strains that carried a new plasmid with a molecular mass of 5.1 MDa was coincidental with an increase in isolation of S. sonnei in 1988. This new plasmid was present in 87.5% of the isolates that were resistant to chloramphenicol. Shigellosis in Guadalajara follows a pattern of hyperendemic transmission with transient peaks of high-frequency isolation of S. sonnei. This pattern results from the concurrent presence of a heterogeneous group of strains as opposed to the widespread transmission of one or a few clones.

  1. Genetic diversity of Mycobacterium tuberculosis complex strains isolated from patients with pulmonary tuberculosis in Anambra State, Nigeria.

    Science.gov (United States)

    Uzoewulu, Gertrude N; Lawson, Lovett; Nnanna, Ibeh S; Rastogi, Nalin; Goyal, Madhu

    2016-03-01

    In this study, we analyzed Mycobacterium tuberculosis complex (MTC) genetic diversity in Anambra State, Nigeria based on spoligotyping followed by 5-loci exact tandem repeats (ETRs). Spoligotyping of 180 MTC strains isolated in 2009-2011 from pulmonary tuberculosis (TB) patients led to a total of 31 distinct patterns. A comparison with the SITVIT2 international database showed that all the 31 patterns could be classified as Shared-types (SITs) in this database; briefly, 26/31 SITs (n=174 isolates) matched a preexisting shared-type in the database, whereas 5/31 SITs (n=6 isolates) were newly created due to 2 or more strains belonging to an identical new pattern within this study (SIT3396) or after a match with an orphan in the database (SIT3397, SIT3398, SIT3399 and SIT3400). A total of 18/31 SITs containing 167 or 92.8% isolates were clustered within this study (2-89 isolates per cluster) while 13/31 SITs contained unique strains. Using VNTR typing, a total of 36 distinct patterns were identified; 27 patterns (n=157 isolates) matched a pattern already reported in the SITVIT2 database. Combination of both the methods generated 47 combined patterns for the 180 strains: 17 belonged to clustered isolates (n=127 isolates or 70.5%) while 30 corresponded to as many unique strains (note 23 strains could not be typed using 5-loci ETRs). No correlation was found between the spoligotyping pattern and the HIV status of the patient or drug sensitivity of the strain. This study showed that the LAM10-CAM prototype SIT61 accounted for highest number of isolates (n=89) in Anambra State, showing its relative contribution to the TB burden in the study.

  2. Isolation and characterization of an atypical Siberian sturgeon herpesvirus strain in Russia: novel North American Acipenserid herpesvirus 2 strain in Europe?

    Science.gov (United States)

    Doszpoly, A; Kalabekov, I M; Breyta, R; Shchelkunov, I S

    2017-02-27

    Siberian sturgeon herpesvirus (SbSHV) was isolated in Russia for the first time in 2006. Nine SbSHV isolates were recovered from different fish hatcheries producing the same cytopathic effect in cell cultures, the same clinical signs and mortality kinetics in virus-infected fish and the same virus neutralization pattern and shared identical nucleotide sequences. In 2011, a new isolate was recovered from juvenile sturgeon, which caused completely different cytopathic effect. That isolate was not readily neutralized by Siberian sturgeon hyperimmune antisera, and its DNA was not recognized by the routine PCR developed for SbSHV detection. Molecular study of the novel isolate revealed that it was more closely related to North American Acipenserid herpesvirus 2 (AciHV-2) isolates from white sturgeon, while the genome sequences of the former SbSHV isolates showed high similarity to the AciHV-2 isolated from shortnose sturgeon. While clinical signs and mortality caused by the novel isolate in infected Siberian sturgeon were similar to those of the formerly described SbSHV isolates, the incubation period and mean time to death produced by the novel isolate were twice as long. The differences between the former isolates and the recent one suggest that a novel SbSHV strain emerged in Europe and the molecular findings imply its North American origin.

  3. Diversity of Saccharomyces cerevisiae strains isolated from Borassus akeassii palm wines from Burkina Faso in comparison to other African beverages.

    Science.gov (United States)

    Tapsoba, François; Legras, Jean-Luc; Savadogo, Aly; Dequin, Sylvie; Traore, Alfred Sababenedyo

    2015-10-15

    In South-West of Burkina Faso, palm wine is produced by spontaneous fermentation of the sap from a specific palm tree Borassus akeassii and plays an important role in people's lives. Saccharomyces cerevisiae is the main agent of this alcoholic fermentation but little is known about the diversity of the isolates from palm. In this work, 39 Saccharomyces cerevisiae strains were isolated from palm wine samples collected from 14 sites in Burkina Faso, as well as 7 isolates obtained from sorghum beer (Dolo) from 3 distant sites. Their diversity was analyzed at 12 microsatellite loci, and compared to the genotypes obtained for other African yeast populations isolated from Cocoa hulks from Ghana, sorghum beer from Ivory Coast, palm wine from Djibouti Republic, and to our database of strains from miscellaneous origins (bread, beer, wine, sake, oaks…). The ploidy of these strains has been assessed as well by flow cytometry. Our results show that B. akeassii palm wine contains a specific yeast population of diploid strains, different from Dolo produced in the same area and from other palm wine strains from Ivory Coast, Nigeria, or Djibouti Republic. In contrast, Dolo strains appeared as a group of related and mainly tetraploid strains despite being isolated from different countries.

  4. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

    Directory of Open Access Journals (Sweden)

    Jinhua Wang

    Full Text Available Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  5. Isolation and characterization of atrazine mineralizing Bacillus subtilis strain HB-6.

    Science.gov (United States)

    Wang, Jinhua; Zhu, Lusheng; Wang, Qi; Wang, Jun; Xie, Hui

    2014-01-01

    Atrazine is a widely used herbicide with great environmental concern due to its high potential to contaminate soil and waters. An atrazine-degrading bacterial strain HB-6 was isolated from industrial wastewater and the 16S rRNA gene sequencing identified HB-6 as a Bacillus subtilis. PCR assays indicated that HB-6 contained atrazine-degrading genes trzN, atzB and atzC. The strain HB-6 was capable of utilizing atrazine and cyanuric acid as a sole nitrogen source for growth and even cleaved the s-triazine ring and mineralized atrazine. The strain demonstrated a very high efficiency of atrazine biodegradation with a broad optimum pH and temperature ranges and could be enhanced by cooperating with other bacteria, suggesting its huge potential for remediation of atrazine-contaminated sites. To our knowledge, there are few Bacillus subtilis strains reported that can mineralize atrazine, therefore, the present work might provide some new insights on atrazine remediation.

  6. Molecular epidemiology and evolution of worldwide enterovirus 71 strains isolated from 1970 to 2004

    Institute of Scientific and Technical Information of China (English)

    DONG XiaoNan; YING Jian; CHEN YingHua

    2007-01-01

    Human enterovirus 71 (EV71) is one of the major etiological agents of the hand-foot-and-mouth disease (HFMD) that often causes severe neurological complications. Recently, its outbreaks mainly take place in the torrid zone of the Asia-Pacific region. To study the evolution and genetic variability, we collected 532 EV71 strains with almost complete or complete VP1 sequences (891 nt) isolated worldwide from 1970 to 2004. The pairwise homologies and genetic distances were analyzed. Most strains belong to previously identified genotype B and C. However, a unique strain R13223-IND-01 appears not to fall into current three genotypes (A, B and C), and probably represents a new genotype D. Some orphan strains were observed in the genotypes B and C, and their significance in the EV71 evolution was discussed.Moreover, there is a significant co-variance of 6 discrete positions on VP1 (amino acid 43, 58, 164, 184,240 and 249). This high co-variability is tightly related with the subgenotypes.

  7. Complete genome analysis of two new bacteriophages isolated from impetigo strains of Staphylococcus aureus.

    Science.gov (United States)

    Botka, Tibor; Růžičková, Vladislava; Konečná, Hana; Pantůček, Roman; Rychlík, Ivan; Zdráhal, Zbyněk; Petráš, Petr; Doškař, Jiří

    2015-08-01

    Exfoliative toxin A (ETA)-coding temperate bacteriophages are leading contributors to the toxic phenotype of impetigo strains of Staphylococcus aureus. Two distinct eta gene-positive bacteriophages isolated from S. aureus strains which recently caused massive outbreaks of pemphigus neonatorum in Czech maternity hospitals were characterized. The phages, designated ϕB166 and ϕB236, were able to transfer the eta gene into a prophageless S. aureus strain which afterwards converted into an ETA producer. Complete phage genome sequences were determined, and a comparative analysis of five designed genomic regions revealed major variances between them. They differed in the genome size, number of open reading frames, genome architecture, and virion protein patterns. Their high mutual sequence similarity was detected only in the terminal regions of the genome. When compared with the so far described eta phage genomes, noticeable differences were found. Thus, both phages represent two new lineages of as yet not characterized bacteriophages of the Siphoviridae family having impact on pathogenicity of impetigo strains of S. aureus.

  8. Isolation and characterization of an Antarctic Flavobacterium strain with agarase and alginate lyase activities

    Directory of Open Access Journals (Sweden)

    Lavín Paris

    2016-09-01

    Full Text Available Several bacteria that are associated with macroalgae can use phycocolloids as a carbon source. Strain INACH002, isolated from decomposing Porphyra (Rhodophyta, in King George Island, Antarctica, was screened and characterized for the ability to produce agarase and alginate-lyase enzymatic activities. Our strain INACH002 was identified as a member of the genus Flavobacterium, closely related to Flavobacterium faecale, using 16S rRNA gene analysis. The INACH002 strain was characterized as psychrotrophic due to its optimal temperature (17ºC and maximum temperature (20°C of growth. Agarase and alginate-lyase displayed enzymatic activities within a range of 10°C to 50°C, with differences in the optimal temperature to hydrolyze agar (50°C, agarose (50°C and alginate (30°C during the first 30 min of activity. Strain Flavobacterium INACH002 is a promising Antarctic biotechnological resource; however, further research is required to illustrate the structural and functional bases of the enzymatic performance observed during the degradation of different substrates at different temperatures.

  9. Isolation of recombinant field strains of Marek's disease virus integrated with reticuloendotheliosis virus genome fragments

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Zhi; CUI; Zhizhong

    2005-01-01

    Two Marek's disease virus (MDV) field strains were isolated from chickens with tumors independently from Guangdong and Guangxi provinces, and it was confirmed that there were no co-infections with reticuloendotheliosis viruses (REV) in chicken embryo fibroblast cells (CEF) in indirect fluorescence antibody test (IFA) with REV-specific monoclonal antibodies. By dot blot hybridization and PCR of genomic DNA of MDV-infected CEF, it was indicated that LTR fragments of REV genome were integrated into genome of these two MDV field strains. To amplify and clone the integrated REV LTR with MDV sequence at the junction, 4 primers from REV LTR and 7 primers from MDV genome fragment with REV LTR insertion hot points were synthesized and 28 (4x7) pairs of primers (one from REV and another from MDV for each pair) were used in PCR while using the genomic DNA of both strains as the templates. The sequence data demonstrated that both recombinant field strains contained the same REV LTR inserted into MDV at the identical sites in US fragment of the genomes. From the above, it was speculated that both recombinant field MDVs were originated from a same recombinant virus and spread among chicken flocks in two provinces.

  10. Technological properties of Lactobacillus plantarum strains isolated from grape must fermentation.

    Science.gov (United States)

    Berbegal, Carmen; Peña, Nuria; Russo, Pasquale; Grieco, Francesco; Pardo, Isabel; Ferrer, Sergi; Spano, Giuseppe; Capozzi, Vittorio

    2016-08-01

    Malolactic fermentation (MLF) is a secondary fermentation in wine that usually takes place during or at the end of alcoholic fermentation. Lactobacillus plantarum is able to conduct MLF (particularly under high pH conditions and in co-inoculation with yeasts), and some strains are commercially used as MLF starter cultures. Recent evidences suggest a further use of selected L. plantarum strains for the pre-alcoholic acidification of grape must. In this study, we have carried out an integrated (molecular, technological, and biotechnological) characterization of L. plantarum strains isolated from Apulian wines in order to combine the two protechnological features (MLF performances and must acidification aptitudes). Several parameters such as sugar, pH and ethanol tolerance, resistance to lyophilisation and behaviour in grape must were evaluated. Moreover, the expression of stress gene markers was investigated and was linked to the ability of L. plantarum strains to grow and perform MLF. Co-inoculation of Saccharomyces cerevisiae and L. plantarum in grape must improves the bacterial adaptation to harsh conditions of wine and reduced total fermentation time. For the first time, we applied a polyphasic approach for the characterization of L. plantarum in reason of the MLF performances. The proposed procedure can be generalized as a standard method for the selection of bacterial resources for the design of MLF starter cultures tailored for high pH must.

  11. Characterization of a virulent Leptospira interrogans strain isolated from an abandoned swimming pool.

    Science.gov (United States)

    Forster, Karine M; Hartwig, Daiane D; Seixas, Fabiana K; McBride, Alan J A; Monte, Leonardo G; Recuero, Ana Lúcia C; Brod, Claudiomar S; Hartleben, Cláudia P; Amaral, Marta; Dellagostin, Odir A

    2013-01-01

    Pathogenic Leptospira spp. are the etiological agents of leptospirosis, an important disease of both humans and animals. In urban settings, L. interrogans serovars are the predominant cause of disease in humans. The purpose of this study was to characterize a novel Leptospira isolate recovered from an abandoned swimming pool. Molecular characterization through sequencing of the rpoB gene revealed 100% identity with L. interrogans and variable-number tandem-repeat (VNTR) analysis resulted in a banding pattern identical to L. interrogans serogroup Icterohaemorrhagiae, serovar Copenhageni or Icterohaemorrhagiae. The virulence of the strain was determined in a hamster model of lethal leptospirosis. The lethal dose 50% (LD50) was calculated to be two leptospires in female hamsters and a histopathological examination of infected animals found typical lesions associated with severe leptospirosis, including renal epithelium degeneration, hepatic karyomegaly, liver-plate disarray and lymphocyte infiltration. This highly virulent strain is now available for use in further studies, especially evaluation of vaccine candidates.

  12. Evolutionary and Ecological Characterization of Mayaro Virus Strains Isolated during an Outbreak, Venezuela, 2010.

    Science.gov (United States)

    Auguste, Albert J; Liria, Jonathan; Forrester, Naomi L; Giamba