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Sample records for subsp simsii alstroemeriaceae

  1. Análisis comparativo del cariotipo en poblaciones de Alstroemeria ligtu subsp. ligtu y A. ligtu subsp. simsii (Alstroemeriaceae de Chile

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    Carlos M. Baeza

    2006-01-01

    Full Text Available Alstroemeria (Alstroemeriaceae es un género endémico de América del Sur. En Chile, este género se distribuye desde el extremo norte hasta la Patagonia, y la mayor diversidad de especies se encuentra en la zona central. Precisamente en esta zona crece Alstroemeria ligtu con sus 3 subespecies: A. ligtu subsp. ligtu, A. ligtu subsp. incarnata, A. ligtu subsp. simsii. Se realizó un estudio comparativo del cariotipo de individuos provenientes de 5 poblaciones de A. ligtu subsp. ligtu de la VIII Región, y de una población de A. ligtu subsp. simsii de la V Región, mediante tinción de los cromosomas con DAPI u orceína acética. Las seis poblaciones estudiadas presentaron un cariotipo asimétrico, con 2n=2x=16 cromosomas. Las poblaciones de A. ligtu subsp. ligtu presentaron una fórmula haploide conformada por cuatro cromosomas metacéntricos (los pares 1 y 2 con microsatélites, uno submetacéntrico con microsatélite y tres telocéntricos con microsatélites. La población de A. ligtu subsp. simsii se caracterizó por poseer cinco cromosomas metacéntricos (el par 2 con un microsatélite y el par 6 con una constricción secundaria y tres cromosomas telocéntricos con satélite. Estos resultados indican que el cariotipo en A. ligtu es variable, y es probable que cambios a nivel cromosómico hayan contribuido en la diversificación de esta especie.

  2. The Karyotype of Alstroemeria diluta Ehr. Bayer subsp. chrysantha (Alstroemeriaceae Karyotype of Alstroemeria diluta Ehr. Bayer subsp. chrysantha (Alstroemeriaceae

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    Carlos M Baeza

    2010-12-01

    Full Text Available The karyotype of Alstroemeria diluta subsp. chrysantha Ehr. Bayer from Chile was examined. The species has 2n = 2x = 16 chromosomes, with 4m + 4sm + 2st-sat + 4t + 2t-sat. The reported karyotype is very asymmetrical (AsK % = 71.4 and Syi = 40.0%. This karyotype is similar to that published previously for Alstroemeria graminea Phil.Alstroemeria diluta subsp. chrysantha Ehr. Bayer (Alstroemeriaceae fue examinada citológicamente. Esta especie presenta un número cromosómico somático de 2n = 2x = 16 cromosomas, con una fórmula haploide constituida por 4m + 4sm + 2st-sat + 4t + 2t-sat cromosomas. El cariotipo es muy asimétrico, con valores de AsK % = 71,4 y Syi = 40,0%. Estos resultados se compararon con los de Alstroemeria graminea Phil., especie que presenta un cariotipo muy similar.

  3. Variabilidad genética y morfológica y estructuración poblacional en Alstroemeria hookeri subsp. hookeri (Alstroemeriaceae, endémica de Chile Genetic and morphological variation and population structure in Alstroemeria hookeri subsp. hookeri (Alstroemeriaceae, endemic to Chile

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    EDUARDO RUIZ

    2010-12-01

    genética intrapoblacional en esta subespecie. Los resultados del AMOVA indican que la variabilidad se da principalmente entre poblaciones. Sin embargo, estos resultados variaron cuando se analizan las poblaciones de ambos rangos de distribución separadamente. La similitud genética no coincide con la divergencia morfológica entre individuos de la costa e interior, indicando que algunas poblaciones del interior están más relacionadas genéticamente, con poblaciones costeras que con aquellas de su mismo rango geográfico.The genus Alstroemeria consists of 82, exclusively South American taxa that are distributed mainly in Chile and Brazil. The great economic relevance, that has acquired the Chilean Alstroemeria species, as ornamentáis, has aroused great interest in the morphological variability of their flowers and genetic variability in those species with potential economic valué. One of these species is Alstroemeria hookeri which has four subspecies, of which A. hookeri subsp. hookeri, is endemic to the Maule and Biobio Regions. Its geographic distribution consists of two ranges, separated by the Coastal Mountain. Thus, there are populations growing in the coastal range, between 5-20 m high, in the provinces of Arauco, Concepción, Nuble, and Cauquenes, and populations of the Central Valley, growing between 100-150 m in the Biobio and Nuble provinces. Preliminary evidence indicated phenotypic differences between coastal and Central Valley populations, principally in colour and shape of the tepals. For this reason, a comparative morphological study was carried out in the complete geographic distribution of this subspecies and a population genetics study was conducted also, especially to determine the levéis of population structure. Thirty three floral characters were analyzed through ordination methods. The morphological study shows a slight tendency to separate the populations into two groups, which coincide with the extremes of morphological variation and both

  4. Alstroemeria presliana Herb. (Alstroemeriaceae in Chile from a Cytogenetic Perspective Alstroemeria presliana Herb. (Alstroemeriaceae en Chile Bajo una Perspectiva Citogenética

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    Carlos M Baeza

    2008-12-01

    Full Text Available Alstroemeria (Alstroemeriaceae is an endemic genus of South America with two major distribution centers in the continent: Chile and Brazil. In Chile the genus is distributed from the North, near Iquique (20º13’ S, 70º09’ W to the Chilean and Argentine Patagonia (53º10’ S, 70º54’ W. The central zone of Chile presents the highest number of species. A. presliana Herb. grows from Curicó (34º59’ S, 71º14’ W to Cautín (38º45’ S, 72º34’ W in Chile and Neuquén (36º50’ S, 71º05’ W, Argentina. A comparative karyotype study was made between a population of A. presliana subsp. presliana and a population of A. presliana subsp. australis Ehr. Bayer. Both populations presented asymmetric karyotypes, with 2n = 2x = 16 chromosomes, but with different chromosome formulae: A. presliana subsp. presliana has a haploid formula with 4m + 1sm-sat + 1st-sat + 2t, i.e., four pairs ofmetacentric chromosomes, one submetacentric pair with satellite, one subtelocentric pair with satellite, and two telocentrics pairs. A. presliana subsp. australis has a formula with 2m + 1m-sat + 1sm + 4t chromosomes, i.e., two pairs of metacentric chromosomes, one metacentric pair with satellite, one submetacentric pair, and four telocentrics chromosomes. These results indicated that the karyotype of the subspecies is very different, and it would be possible to recognize A. presliana subsp. australis as a new species.Alstroemeria (Alstroemeriaceae es un género endémico de Sudamérica y presenta dos grandes centros de distribución: Chile y Brasil. En Chile se distribuye desde el norte, cerca de Iquique (20º13’ S, 70º09’ O hasta la Patagonia (53º10’ S, 70º54’ O. La zona central del país presenta el mayor número de especies. A. presliana Herb. crece con una distribución que va desde Curicó (34º59’ S, 71º14’ O a Cautín (38º45’ S, 72º34’ O en Chile y en la Provincia de Neuquén (36º50’ S, 71º05’ O, Argentina. Se hizo un

  5. Alstroemeriaceae endémicas del Perú

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    Blanca León

    2013-10-01

    Full Text Available La familia Alstroemeriaceae es reconocida en el Perú con dos géneros, Alstroemeria y Bomarea, y 85 especies (Brako & Zarucchi, 1993; Ulloa Ulloa et al., 2004, básicamente hierbas y enredaderas. En este trabajo reconocemos 22 endemismos en el género Bomarea. Estos taxones endémicos ocupan varias regiones ecológicas, incluyendo la del Bosque Húmedo Montano y de la Puna Seca y Húmeda, entre los 1000 y 4600 m de altitud. Aproximadamente un tercio de esas especies, son conocidas solamente de sus localidades tipo, ubicadas en cadenas montañosas aisladas o en cuencas que han recibido escasa herborización los últimos 80 años, como la del Monzón. Nueve de estas especies están representadas en áreas naturales protegidas.

  6. Alstroemeriaceae en Perú y áreas vecinas

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    Anton Hofreiter

    2006-10-01

    Full Text Available Las Alstroemeriaceae peruanas fueron revisadas por última vez por Killip (1936. Es necesaria una nueva revisión. Cinco viajes de campo de varias semanas cada uno fueron emprendidos a Sudamérica con el fin de estudiar las plantas in situ. En el presente trabajo se describe la historia taxonómica y colección de los géneros con especial énfasis en el Perú. El área descrita no considera fronteras políticas sino unidades geográficas de acuerdo a Baumann (1988, Berry (1982, Duellman (1979, Simpson (1975, 1979 y Weigend (2002. Se reconocen cinco unidades geográficas: Región Amotape-Huancabamba (Ecuador, Perú, Cordillera Occidental (Perú, Cordillera Central (Perú, Cordillera Oriental (Bolivia, Perú y el Altiplano (Bolivia, Perú. Se brinda una clave taxonómica en inglés y español para determinar las especies del Perú. Para cada una de las especies se discute la forma típica de crecimiento y su variabilidad, preferencias de hábitat y distribución general. Se identifican las especies de Ruiz & Pavón (1802. Ellos describieron en su Flora de Chile y Perú 23 especies de Alstroemeria, 18 fueron de Perú, ahora 17 son incluidas en Bomarea, todas proceden de Perú. El género Bomarea está subdividido en 4 subgéneros: Baccata, Bomarea s. str., Sphaerine y Winchurea (Hofreiter & Tillich, 2002. Alstroemeria no es dividido en subgéneros, pero existen dos grupos reconocidos, Alstroemeria de Chile y Brasil. En el área de estudio se encuentran dos especies de Alstroemeria y 68 de Bomarea, de ellas 43 especies pertenecen al subgénero Bomarea, 9 especies al subgénero Sphaerine y 16 especies al subgénero Wichuraea. El subgénero Baccata no se encuentra en el área de estudio. Seis de las especies del género Bomarea son nuevas para la ciencia: B. amazonica, B. libertadensis, B. lopezii, B. macusanii, B. pseudopurpurea, B. weigendii.

  7. Radiation induced chimeric rearrangement flower structure of Rhododendron simsii Planch. (Azaleaindica L.) Use of recurrent irradiation

    International Nuclear Information System (INIS)

    Loose, R. de

    1979-01-01

    A radiation-induced chimeric flower colour sport of vegetatively propagated Rhododendron simsii Planch was recurrently irradiated (up to three times in three consecutive years) with soft X-rays (50kV-30mA), as compared to a single treatment. Because of the low true flower colour mutation frequency the efficiency of the different radiation treatments was compared on the basis of the number of chimeric rearrangements in flower structure i.e. the flower colour change from red with broad white edge towards either homogeneous carminered or white. It is quite clear that recurrent irradiation with appropiate doses is most efficient. (Auth.)

  8. Thymus vulgaris subsp. mansanetianus subsp. nov. (Lamiaceae)

    OpenAIRE

    Ferrer-Gallego, Pedro Pablo; Navarro Peris, Albert Josep; Laguna Lumbreras, Emilio; Mateo Sanz, Gonzalo

    2013-01-01

    RESUMEN: Se describe una nueva subespecie de Thymus vulgaris L. (Lamiaceae); Th. vulgaris subsp. mansanetianus subsp. nov., caracterizada por presentar un hábito postrado, tallos estoloníferos, decumbentes y radicantes, hojas muy estrechas y una floración otoñal. ABSTRACT: Thymus vulgaris subsp. mansanetianus subsp. nov. (Lamiaceae). A new subspecies of Thymus vulgaris L. (Lamiaceae); Th. vulgaris subsp. mansanetianus subsp. nov. is described. This new subspecies is characterized by its prost...

  9. Leaf trichomes and foliar chemistry mediate defence\\ud against glasshouse thrips; Heliothrips haemorrhoidalis\\ud (Bouché) in Rhododendron simsii

    OpenAIRE

    Scott-Brown, Alison S.; Gregory, Tom; Farrell, Iain W.; Stevenson, Philip C.

    2016-01-01

    Herbivore defence mechanisms are a costly diversion of resources away from growth and reproduction. Thus time-limited and tissue specific expression in critical plant parts is more efficient as defined by optimal defence theory. Surprisingly little is known about Rhododendron herbivore defence but it may be mediated by combined chemical and physical mechanisms. Rhododendron simsii Planch. survives cyclic infestations of a leaf-feeding thrips, Heliothrips haemorrhoidalis, which severely damage...

  10. Leaf fossils of Luzuriaga and a monocot flower with in situ pollen of Liliacidites contortus Mildenh. & Bannister sp. nov. (Alstroemeriaceae) from the Early Miocene.

    Science.gov (United States)

    Conran, John G; Bannister, Jennifer M; Mildenhall, Dallas C; Lee, Daphne E; Chacón, Juliana; Renner, Susanne S

    2014-01-01

    The Foulden Maar lake sediments in Otago, South Island, New Zealand, date to the earliest Miocene and provide an important picture of the diversity of the Australasian biota, paleoecology, and climate at a time when New Zealand had a smaller land area than today. The diverse rainforest contains many taxa now restricted to Australia, New Caledonia, or South America. The presence of Luzuriaga-like fossils in these deposits is important for understanding Alstroemeriaceae evolution and the biogeography of genera shared between New Zealand and South America. Leaves and a flower with in situ pollen that resemble extant Luzuriaga are described and placed phylogenetically. Geographic range information and a molecular clock model for the Alstroemeriaceae were used to investigate possible biogeographic scenarios and the influence of the new fossil on inferred divergence times. Luzuriaga peterbannisteri Conran, Bannister, Mildenh., & D.E.Lee sp. nov. represents the first macrofossil record for Alstroemeriaceae. An associated Luzuriaga-like flower with in situ fossil pollen of Liliacidites contortus Mildenh. & Bannister sp. nov. is also described. The biogeographic analysis suggests that there have been several dispersal events across the Southern Ocean for the genus, with the fossil representing a now-extinct New Zealand lineage. Luzuriaga was present in Early Miocene New Zealand, indicating a long paleogeographic history for the genus, and L. peterbannisteri strengthens biogeographic connections between South America and Australasia during the Oligocene and earliest Miocene.

  11. Protective Effect and Mechanism of Total Flavones from Rhododendron simsii Planch Flower on Cultured Rat Cardiomyocytes with Anoxia and Reoxygenation

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    Yi Jiao

    2015-01-01

    Full Text Available Many flavonoids have cardioprotection against myocardial ischemia/reperfusion (I/R injury. Total flavones from Rhododendron simsii Planch flower (TFR can protect myocardial ischemic injuries. However, its protective mechanism is still unknown. The present study was designed to investigate the mechanism of TFR on myocardial I/R and anoxia/reoxygenation (A/R injuries. Rat model of myocardial I/R injury was made, and myocardial infarction was determined. A/R injury was induced in cultured rat cardiomyocytes; cellular damage was evaluated by measuring cell viability, LDH and cTnT releases, and MDA content. Expressions of ROCK1 and ROCK2 protein were examined by Western blot analysis, and K+ currents were recorded by using whole-cell patch clamp technique. TFR 20~80 mg/kg markedly reduced I/R-induced myocardial infarction. TFR 3.7~300 mg/L significantly inhibited A/R-induced reduction of cell viability, LDH and cTnT releases, and MDA production. Exposure to A/R significantly increased ROCK1 and ROCK2 expressions in rat cardiomyocytes, but TFR 33.3~300 mg/L obviously inhibited this increase. 300 mg/L TFR significantly augmented inward rectifier K+ current and other K+ currents in rat cardiomyocytes. These results indicate that TFR has a protective effect on rat cardiomyocytes A/R damage, and the protective mechanism may be engaged with the inhibition of ROCK1 and ROCK2 and activation of K+ channels.

  12. Multipoint-likelihood maximization mapping on 4 segregating populations to achieve an integrated framework map for QTL analysis in pot azalea (Rhododendron simsii hybrids

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    Van Bockstaele Erik

    2010-01-01

    Full Text Available Abstract Background Azalea (Rhododendron simsii hybrids is the most important flowering pot plant produced in Belgium, being exported world-wide. In the breeding program, flower color is the main feature for selection, only in later stages cultivation related plant quality traits are evaluated. As a result, plants with attractive flowering are kept too long in the breeding cycle. The inheritance of flower color has been well studied; information on the heritability of cultivation related quality traits is lacking. For this purpose, QTL mapping in diverse genetic backgrounds appeared to be a must and therefore 4 mapping populations were made and analyzed. Results An integrated framework map on four individual linkage maps in Rhododendron simsii hybrids was constructed. For genotyping, mainly dominant scored AFLP (on average 364 per population and MYB-based markers (15 were combined with co-dominant SSR (23 and EST markers (12. Linkage groups were estimated in JoinMap. A consensus grouping for the 4 mapping populations was made and applied in each individual mapping population. Finally, 16 stable linkage groups were set for the 4 populations; the azalea chromosome number being 13. A combination of regression mapping (JoinMap and multipoint-likelihood maximization (Carthagène enabled the construction of 4 maps and their alignment. A large portion of loci (43% was common to at least two populations and could therefore serve as bridging markers. The different steps taken for map optimization and integration into a reference framework map for QTL mapping are discussed. Conclusions This is the first map of azalea up to our knowledge. AFLP and SSR markers are used as a reference backbone and functional markers (EST and MYB were added as candidate genes for QTL analysis. The alignment of the 4 maps on the basis of framework markers will facilitate in turn the alignment of QTL regions detected in each of the populations. The approach we took is

  13. [Streptococcus equi subsp. Zooepidemicus endocarditis].

    Science.gov (United States)

    Villamil, Iago; Serrano, Marta; Prieto, Elisabeth

    2015-04-01

    Zoonotic infections with S. equi subsp. zooepidemicus have been reported occasionally in individuáis with exposure to farm animals, especially horses. They exceptionally present as a human pathogen. We present a S. equi subsp. zooepidemicus well documented endocarditis after a horse's female genital tract contact, and excellent surveillance with exclusive antibiotic management.

  14. Taxonomic relatedness between Pectobacterium carotovorum subsp. carotovorum, Pectobacterium carotovorum subsp. odoriferum and Pectobacterium carotovorum subsp. brasiliense subsp. nov.

    Science.gov (United States)

    Nabhan, S; De Boer, S H; Maiss, E; Wydra, K

    2012-10-01

    Pectobacterium carotovorum is a heterogeneous species consisting of two named subspecies, P. carotovorum subsp. carotovorum and P. carotovorum subsp. odoriferum. A third subspecies, P. carotovorum subsp. brasiliense, was previously proposed. The study aimed to confirm the subspecies status and validate the proposed name of P. carotovorum subsp. brasiliense using a novel and standard microbial taxonomy. DNA-DNA hybridization confirmed that P. carotovorum subsp. brasiliense is a different species from P. wasabiae, P. betavasculorum and P. atrosepticum, with 28, 35 and 55% similarity values, respectively, but is a member of the P. carotovorum species with 73-77% similarity values. Sequencing the entire 16S rRNA gene of two polymorphic copies from strains of each of the P. carotovorum subspecies demonstrated that the average 16S rRNA gene sequence diversity between P. carotovorum subsp. brasiliense and P. carotovorum subsp. carotovorum was lower than the maximum genetic distances between two sequence types obtained from the same strain. Multilocus sequence analysis based on eight housekeeping genes (mtlD, acnA, icdA, mdh, pgi, gabA, proA and rpoS) differentiated the subspecies and delineated two P. carotovorum subsp. brasiliense clades. Pectobacterium carotovorum subsp. brasiliense clade I was comprised of strains isolated from Brazil and Peru, while clade II included strains from Asia, North America and Europe. Strains in clade I but not clade II were phenotypically consistent with the original description of P. carotovorum subsp. brasiliense in that they produced reducing substances from sucrose and acid from α-methyl glucoside. The type strain for P. carotovorum subsp. brasiliense 212(T) (= LMG2137(T) = IBSBF1692(T) = CFBP6617(T) ) was previously designated. The GC mol content of the type strain is 51·7%. the study introduces a full description for the strains belonging to the two different clades assigned to P. carotovorum subsp. brasiliense.

  15. Identification of Mycobacterium avium subsp. hominissuis Isolated From Drinking Water

    Science.gov (United States)

    Mycobacterium avium (MA) is divided into four subspecies based primarily on host-range and consists of MA subsp. avium (birds), MA subsp. silvaticum (wood pigeons), MA subsp. paratuberculosis (broad, poorly-defined host range), and the recently described MA subsp. hominissuis (hu...

  16. Transfer of several phytopathogenic Pseudomonas species to Acidovorax as Acidovorax avenae subsp. avenae subsp. nov., comb. nov., Acidovorax avenae subsp. citrulli, Acidovorax avenae subsp. cattleyae, and Acidovorax konjaci.

    Science.gov (United States)

    Willems, A; Goor, M; Thielemans, S; Gillis, M; Kersters, K; De Ley, J

    1992-01-01

    DNA-rRNA hybridizations, DNA-DNA hybridizations, polyacrylamide gel electrophoresis of whole-cell proteins, and a numerical analysis of carbon assimilation tests were carried out to determine the relationships among the phylogenetically misnamed phytopathogenic taxa Pseudomonas avenae, Pseudomonas rubrilineans, "Pseudomonas setariae," Pseudomonas cattleyae, Pseudomonas pseudoalcaligenes subsp. citrulli, and Pseudomonas pseudoalcaligenes subsp. konjaci. These organisms are all members of the family Comamonadaceae, within which they constitute a separate rRNA branch. Only P. pseudoalcaligenes subsp. konjaci is situated on the lower part of this rRNA branch; all of the other taxa cluster very closely around the type strain of P. avenae. When they are compared phenotypically, all of the members of this rRNA branch can be differentiated from each other, and they are, as a group, most closely related to the genus Acidovorax. DNA-DNA hybridization experiments showed that these organisms constitute two genotypic groups. We propose that the generically misnamed phytopathogenic Pseudomonas species should be transferred to the genus Acidovorax as Acidovorax avenae and Acidovorax konjaci. Within Acidovorax avenae we distinguished the following three subspecies: Acidovorax avenae subsp. avenae, Acidovorax avenae subsp. cattleyae, and Acidovorax avenae subsp. citrulli. Emended descriptions of the new taxa are presented.

  17. Respon Fagositosis Leukosit Polimorf Babi (in vitro Terhadap Streptoccocus equi Subsp. Zooepidemicus (PHAGOCYTIC RESPONSE OF SWINE POLYMORPH LEUCOCYTES (IN VITRO TO STREPTOCCOCUS EQUI SUBSP. ZOOEPIDEMICUS

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    Iwan Harjono Utama

    2016-08-01

    Full Text Available Respon Fagositosis Leukosit Polimorf Babi (in vitro Terhadap Streptoccocus equi Subsp. Zooepidemicus (PHAGOCYTIC RESPONSE OF SWINE POLYMORPH LEUCOCYTES (IN VITRO TO STREPTOCCOCUS EQUI SUBSP. ZOOEPIDEMICUS

  18. Growth potential of Eucalyptus globulus subsp. bicostata ...

    African Journals Online (AJOL)

    With forestry expanding into mid-altitude drier and warmer, or drier and colder, sites the search to identify reasonably well-performing species/provenances for such ... susceptible than other provenances and controls, also outperformed other E. globulus subsp. bicostata provenances for dbh, height, basal area and volume.

  19. Rapid detection of Mycobacterium avium subsp. paratuberculosis ...

    African Journals Online (AJOL)

    Therefore, alternative diagnostic tests such as PCR, are needed for quick detection of infected animals. In this study, the conventional enrichment and isolation procedure and two IS900-based PCR methods for detection of Mycobactrium avium subsp. paratuberculosis in clinical samples from zoo animals and cattle were ...

  20. Taxonomy of the Anginosus group of the genus Streptococcus and description of Streptococcus anginosus subsp. whileyi subsp. nov. and Streptococcus constellatus subsp. viborgensis subsp. nov.

    Science.gov (United States)

    Jensen, Anders; Hoshino, Tomonori; Kilian, Mogens

    2013-07-01

    The Anginosus group of the genus Streptococcus has been the subject of much taxonomic confusion, which has hampered the full appreciation of its clinical significance. The purpose of this study was to critically re-examine the taxonomy of the Anginosus group, with special attention to β-haemolytic, Lancefield group C strains, using multilocus sequence analysis (MLSA) combined with 16S rRNA gene sequence and phenotypic analyses. Phylogenetic analysis of concatenated sequences of seven housekeeping genes previously used for examination of viridans streptococci distinguished seven distinct and coherent clusters in the Anginosus group. Analyses of 16S rRNA gene sequences and phenotypic characters supported the MLSA clustering and currently recognized taxa of the Anginosus group. Single gene analyses showed considerable allele sharing between species, thereby invalidating identification based on single-locus sequencing. Two novel clusters of β-haemolytic, Lancefield group C strains within the Streptococcus constellatus and Streptococcus anginosus species and isolated from patients with sore throat showed sufficient phylogenetic distances from other clusters to warrant status as novel subspecies. The novel cluster within S. anginosus was identified as the previously recognized DNA homology cluster, DNA group 2. The names S. anginosus subsp. whileyi subsp. nov. (type strain CCUG 39159(T) = DSM 25818(T) = SK1267(T)) and S. constellatus subsp. viborgensis subsp. nov. (type strain SK1359(T) = CCUG 62387(T) = DSM 25819(T)) are proposed.

  1. The first closed genome sequence of Campylobacter fetus subsp. venerealis biovar intermedius

    Science.gov (United States)

    Campylobacter fetus venerealis biovar intermedius is a variant of Campylobacter fetus subsp. venerealis, the causative agent of Bovine Genital Campylobacteriosis. In contrast to Campylobacter fetus subsp. venerealis which is restricted to the genital tract of cattle, Campylobacter fetus subsp. vener...

  2. Biocontrol of Pectobacterium carotovorum subsp. carotovorum using bacteriophage PP1.

    Science.gov (United States)

    Lim, Jeong-A; Jee, Samnyu; Lee, Dong Hwan; Roh, Eunjung; Jung, Kyusuk; Oh, Changsik; Heu, Sunggi

    2013-08-01

    Pectobacterium carotovorum subsp. carotovorum (formerly Erwinia carotovora subsp. carotovora) is a plant pathogen that causes soft rot and stem rot diseases in several crops, including Chinese cabbage, potato, and tomato. To control this bacterium, we isolated a bacteriophage, PP1, with lytic activity against P. carotovorum subsp. carotovorum. Transmission electron microscopy revealed that the PP1 phage belongs to the Podoviridae family of the order Caudovirales, which exhibit icosahedral heads and short non-contractile tails. PP1 phage showed high specificity for P. carotovorum subsp. carotovorum, and several bacteria belonging to different species and phyla were resistant to PP1. This phage showed rapid and strong lytic activity against its host bacteria in liquid medium and was stable over a broad range of pH values. Disease caused by P. carotovorum subsp. carotovorum was significantly reduced by PP1 treatment. Overall, PP1 bacteriophage effectively controls P. carotovorum subsp. carotovorum.

  3. Staphylococcus petrasii subsp. pragensis subsp. nov., occurring in human clinical material.

    Science.gov (United States)

    Švec, Pavel; De Bel, Annelies; Sedláček, Ivo; Petráš, Petr; Gelbíčová, Tereza; Černohlávková, Jitka; Mašlanˇová, Ivana; Cnockaert, Margo; Varbanovová, Ivana; Echahidi, Fedoua; Vandamme, Peter; Pantuček, Roman

    2015-07-01

    Seven coagulase-negative, oxidase-negative and novobiocin-susceptible staphylococci assigned tentatively as Staphylococcus petrasii were investigated in this study in order to elucidate their taxonomic position. All strains were initially shown to form a genetically homogeneous group separated from remaining species of the genus Staphylococcus by using a repetitive sequence-based PCR fingerprinting with the (GTG)5 primer. Phylogenetic analysis based on 16S rRNA gene, hsp60, rpoB, dnaJ, gap and tuf sequences showed that the group is closely related to Staphylococcus petrasii but separated from the three hitherto known subspecies, S. petrasii subsp. petrasii, S. petrasii subsp. croceilyticus and S. petrasii subsp. jettensis. Further investigation using automated ribotyping, MALDI-TOF mass spectrometry, fatty acid methyl ester analysis, DNA-DNA hybridization and extensive biotyping confirmed that the analysed group represents a novel subspecies within S. petrasii, for which the name Staphylococcus petrasii subsp. pragensis subsp. nov. is proposed. The type strain is NRL/St 12/356(T) ( = CCM 8529(T) = LMG 28327(T)).

  4. Iridoid Glucosides from Lamium garganicum subsp. laevigatum

    DEFF Research Database (Denmark)

    Tayfun, Ersöz; Kaya, Duygu; Yalcin, Funda Nuray

    2007-01-01

    Phytochemical investigations on the above ground parts of Lamium garganicum subsp. laevigatum resulted in the isolation of seven iridoid glucosides, shanzhiside methyl ester (1), barlerin (8-O-acetylshanzhiside methyl ester; 2), 6-O-syringyl-8-O-acetylshanzhiside methyl ester (3), 6β-hydroxyipola......Phytochemical investigations on the above ground parts of Lamium garganicum subsp. laevigatum resulted in the isolation of seven iridoid glucosides, shanzhiside methyl ester (1), barlerin (8-O-acetylshanzhiside methyl ester; 2), 6-O-syringyl-8-O-acetylshanzhiside methyl ester (3), 6β......-hydroxyipolamiide (4), lamalbide (5), dehydropenstemoside (6), and sesamoside (7). The structure of the iridoids was elucidated by spectroscopic (UV, IR, 1D- and 2D-NMR, and ESI-MS) evidence....

  5. Campylobacter pinnipediorum sp. nov., isolated from pinnipeds, comprising Campylobacter pinnipediorum subsp. pinnipediorum subsp. nov. and Campylobacter pinnipediorum subsp. caledonicus subsp. nov.

    Science.gov (United States)

    Gilbert, Maarten J; Miller, William G; Leger, Judy St; Chapman, Mary H; Timmerman, Arjen J; Duim, Birgitta; Foster, Geoffrey; Wagenaar, Jaap A

    2017-06-01

    During independent diagnostic screenings of otariid seals in California (USA) and phocid seals in Scotland (UK), Campylobacter-like isolates, which differed from the established taxa of the genus Campylobacter, were cultured from abscesses and internal organs of different seal species. A polyphasic study was undertaken to determine the taxonomic position of these six isolates. The isolates were characterized by 16S rRNA gene and AtpA sequence analysis and by conventional phenotypic testing. The whole-genome sequences were determined for all isolates, and the average nucleotide identity (ANI) was determined. The isolates formed a separate phylogenetic clade, divergent from all other taxa of the genus Campylobacter and most closely related to Campylobactermucosalis. Although all isolates showed 100 % 16S rRNA gene sequence homology, AtpA and ANI analyses indicated divergence between the otariid isolates from California and the phocid isolates from Scotland, which warrants subspecies status for each clade. The two subspecies could also be distinguished phenotypically on the basis of catalase activity. This study shows clearly that the isolates obtained from pinnipeds represent a novel species within the genus Campylobacter, for which the name Campylobacter pinnipediorum sp. nov. is proposed. Within this novel species, the Californian isolates represent a separate subspecies, for which the name C. pinnipediorum subsp. pinnipediorum subsp. nov. is proposed. The type strain for both this novel species and subspecies is RM17260T (=LMG 29472T=CCUG 69570T). The Scottish isolates represent another subspecies, for which the name C. pinnipediorum subsp. caledonicus subsp. nov. is proposed. The type strain of this subspecies is M302/10/6T (=LMG 29473T=CCUG 68650T).

  6. Fragrance components of Platanthera bifolia subsp. osca.

    Science.gov (United States)

    D'Auria, Maurizio; Lorenz, Richard; Racioppi, Rocco; Romano, Vito Antonio

    2017-07-01

    SPME-GC-MS analysis of the scent of Platanthera bifolia subsp. osca collected during the night showed as main components lilac alcohols B, C and D and lilac aldehydes A, B and C. Other significant chemical components were linalool and caryophyllene. Some differences were found in comparison with previously reported analyses of the scent of P. bifolia and Platanthera chlorantha. The most important difference found was in the composition of the ester fraction.

  7. Bacteriological and Molecular Detection of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in Equines of Northern India.

    Science.gov (United States)

    Mir, Irfan Ahmad; Kumar, Bablu; Taku, Anil; Faridi, Farah; Bhat, Mohd Altaf; Baba, Naseer Ahmad; Maqbool, Tahir

    2013-01-01

    Present study was undertaken to study the prevalence of β-haemolytic streptococci in equine of northern temperate region of Jammu and Kashmir, India. One hundred and forty one samples were collected in duplicate from nasopharyngeal tract of diseased (53) and apparently healthy equine (88) for isolation and direct PCR. A total of 77 isolates of streptococci were recovered from 141 samples with an overall prevalence rate of 54.60%. Out of these 77 isolates, 52 were from diseased and 25 from apparently healthy animals. Of the 77 isolates, 4 were identified as Streptococcus equi subsp. equi, 56 as S. equi subsp. zooepidemicus and 17 as S. dysgalactiae subsp. equisimilis. Thus the overall prevalence of S. equi subsp. equi, S. equi subsp. zooepidemicus and S. dysgalactiae subsp. equisimilis was 2.83, 39.71 and 12.05% respectively. The sensitivity of the PCR for the detection of S. equi species was found higher when attempted from direct swab samples.

  8. Epipactis helleborine (L. Crantz subsp. moratoria A. Riechelmann

    Directory of Open Access Journals (Sweden)

    Lipovšek Matej

    2017-06-01

    Full Text Available The group of Epipactis helleborine s.l. includes several subspecies. A new subspecies Epipactis helleborine subsp. moratoria was determined in Slovenia in 2015. It thrives in mixed wood in a region of Gorica at Raztez. Morphologic and phenomenological comparison confirmed clear differences between E. helleborine subsp. moratoria and E. helleborine subsp. helleborine. The characteristic differences seen in E. helleborine subsp. moratoria are the stem which is more or less bent at the level of the leaf base, the plants are smaller and more slender than E. helleborine, there are also differences in the flowers and the leaves. The ovary in E. moratoria is often in a horizontal position, especially at the time of fruiting, wheras in E. helleborine ovary usually hangs down. All of the wild orchids in Slovenia are protected species and among them it is Epipactis helleborine subsp. moratoria which, up till now is only known at one site.

  9. Update on Streptococcus equi subsp equi infections.

    Science.gov (United States)

    Mallicote, Martha

    2015-04-01

    There are few diseases that ignite as much fervor among horse owners as strangles. Streptococcus equi subsp equi (strangles) infections frequently require the treating veterinarian to manage not only the clinical cases but also the biosecurity and provision of information to all involved parties. Although the disease is typically characterized by low mortality and high morbidity, restrictions of horse movement that result from appropriate quarantine procedures often frustrate the involved parties. The aims of this article are to provide clinically relevant information for diagnosis, treatment, and biosecurity management of strangles infection. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Use of PCR-Based Methods for Rapid Differentiation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis

    Science.gov (United States)

    Torriani, Sandra; Zapparoli, Giacomo; Dellaglio, Franco

    1999-01-01

    Two PCR-based methods, specific PCR and randomly amplified polymorphic DNA PCR (RAPD-PCR), were used for rapid and reliable differentiation of Lactobacillus delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis. PCR with a single combination of primers which targeted the proline iminopeptidase (pepIP) gene of L. delbrueckii subsp. bulgaricus allowed amplification of genomic fragments specific for the two subspecies when either DNA from a single colony or cells extracted from dairy products were used. A numerical analysis of the RAPD-PCR patterns obtained with primer M13 gave results that were consistent with the results of specific PCR for all strains except L. delbrueckii subsp. delbrueckii LMG 6412T, which clustered with L. delbrueckii subsp. lactis strains. In addition, RAPD-PCR performed with primer 1254 provided highly polymorphic profiles and thus was superior for distinguishing individual L. delbrueckii strains. PMID:10508059

  11. Genome sequencing identifies Listeria fleischmannii subsp. coloradonensis subsp. nov., isolated from a ranch.

    Science.gov (United States)

    den Bakker, Henk C; Manuel, Clyde S; Fortes, Esther D; Wiedmann, Martin; Nightingale, Kendra K

    2013-09-01

    Twenty Listeria-like isolates were obtained from environmental samples collected on a cattle ranch in northern Colorado; all of these isolates were found to share an identical partial sigB sequence, suggesting close relatedness. The isolates were similar to members of the genus Listeria in that they were Gram-stain-positive, short rods, oxidase-negative and catalase-positive; the isolates were similar to Listeria fleischmannii because they were non-motile at 25 °C. 16S rRNA gene sequencing for representative isolates and whole genome sequencing for one isolate was performed. The genome of the type strain of Listeria fleischmannii (strain LU2006-1(T)) was also sequenced. The draft genomes were very similar in size and the average MUMmer nucleotide identity across 91% of the genomes was 95.16%. Genome sequence data were used to design primers for a six-gene multi-locus sequence analysis (MLSA) scheme. Phylogenies based on (i) the near-complete 16S rRNA gene, (ii) 31 core genes and (iii) six housekeeping genes illustrated the close relationship of these Listeria-like isolates to Listeria fleischmannii LU2006-1(T). Sufficient genetic divergence of the Listeria-like isolates from the type strain of Listeria fleischmannii and differing phenotypic characteristics warrant these isolates to be classified as members of a distinct infraspecific taxon, for which the name Listeria fleischmannii subsp. coloradonensis subsp. nov. is proposed. The type strain is TTU M1-001(T) ( =BAA-2414(T) =DSM 25391(T)). The isolates of Listeria fleischmannii subsp. coloradonensis subsp. nov. differ from the nominate subspecies by the inability to utilize melezitose, turanose and sucrose, and the ability to utilize inositol. The results also demonstrate the utility of whole genome sequencing to facilitate identification of novel taxa within a well-described genus. The genomes of both subspecies of Listeria fleischmannii contained putative enhancin genes; the Listeria fleischmannii subsp

  12. Genetic Diversity of Pectobacterium carotovorum subsp. brasiliensis Isolated in Korea

    Directory of Open Access Journals (Sweden)

    Dong Hwan Lee

    2014-06-01

    Full Text Available The plant pathogenic bacterial genus Pectobacteirum consists of heterogeneous strains. The P. carotovorum species is a complex strain showing divergent characteristics, and a new subspecies named P. carotovorum subsp. brasiliensis has been identified recently. In this paper, we re-identified the P. carotovorum subsp. brasiliensis isolates from those classified under the subspecies carotovorum and newly isolated P. carotovorum subsp. brasiliensis strains. All isolates were able to produce plant cell-wall degrading enzymes such as pectate lyase, polygalacturonase, cellulase and protease. We used genetic and biochemical methods to examine the diversity of P. carotovorum subsp. brasiliensis isolates, and found genetic diversity within the brasiliensis subsp. isolates in Korea. The restriction fragment length polymorphism analysis based on the recA gene revealed a unique pattern for the brasiliensis subspecies. The Korean brasiliensis subsp. isolates were divided into four clades based on pulsed-field gel electrophoresis. However, correlations between clades and isolated hosts or year could not be found, suggesting that diverse brasiliensis subsp. isolates existed.

  13. Genetic Diversity of Pectobacterium carotovorum subsp. brasiliensis Isolated in Korea.

    Science.gov (United States)

    Lee, Dong Hwan; Kim, Jin-Beom; Lim, Jeong-A; Han, Sang-Wook; Heu, Sunggi

    2014-06-01

    The plant pathogenic bacterial genus Pectobacteirum consists of heterogeneous strains. The P. carotovorum species is a complex strain showing divergent characteristics, and a new subspecies named P. carotovorum subsp. brasiliensis has been identified recently. In this paper, we re-identified the P. carotovorum subsp. brasiliensis isolates from those classified under the subspecies carotovorum and newly isolated P. carotovorum subsp. brasiliensis strains. All isolates were able to produce plant cell-wall degrading enzymes such as pectate lyase, polygalacturonase, cellulase and protease. We used genetic and biochemical methods to examine the diversity of P. carotovorum subsp. brasiliensis isolates, and found genetic diversity within the brasiliensis subsp. isolates in Korea. The restriction fragment length polymorphism analysis based on the recA gene revealed a unique pattern for the brasiliensis subspecies. The Korean brasiliensis subsp. isolates were divided into four clades based on pulsed-field gel electrophoresis. However, correlations between clades and isolated hosts or year could not be found, suggesting that diverse brasiliensis subsp. isolates existed.

  14. [Resistance of Lactobacillus casei subsp. casei SY13 and Lactobacillus delbrueckii subsp. bulgaricus LJJ to reactive oxygen species].

    Science.gov (United States)

    Zhang, Shuwen; Lv, Jiaping; Menghe, Bilige; Zhang, Heping; Zhang, Liyu; Song, Jinhui; Wang, Zhifei

    2009-02-01

    We evaluated antioxidative effect of two antioxidative strains, isolated from the traditional fermented dairy products. Both intact cells and cell-free extract of Lactobacillus casei subsp. casei SY13 and Lactobacillus delbrueckii subsp. bulgaricus LJJ were used to study the inhibited effect of linoleic acid peroxidation, the ability of scavenging 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, superoxide anion radical,the ability of tolerancing hydrogen peroxide and the chelating capacity of ferrous ion and reducting activity. Lactobacillus casei subsp. casei SY13 and Lactobacillus delbrueckii subsp. bulgaricus LJJ demonstrated highest inhibition on linoleic acid peroxidation by 62.95% and 66.16%, respectively. The cell-free extract showed excellent scavenging superoxide anion and hydroxyl radicals activity. However, the intact cells of Lactobacillus delbrueckii subsp. bulgaricus LJJ scavenging superoxide and hydroxyl radicals capacity were not detected. The intact cells of Lactobacillus casei subsp. casei SY13 and Lactobacillus delbrueckii subsp. bulgaricus LJJ on 1,1-diphenyl-2-picrylhydrazyl radical scavenging ability and chelating ferrous ion capacity were superior to cell-free extract. The highest reduced activety was equivalent to 305 micromol/L and 294 micromol/L L-cysteine. Two latobacilli strains had good antioxidant capacity. As potential probiotics, it can be used in future.

  15. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines.

    Science.gov (United States)

    Javed, R; Taku, A K; Gangil, Rakhi; Sharma, R K

    2016-08-01

    The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra), characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease) and apparently healthy animals and were sent to laboratory. The organisms were isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and confirmed by cultural characteristics and biochemical tests. Molecular detection of Streptococcus was done directly from cultures using sodA and seM gene-based polymerase chain reaction (PCR). Antibiogram was performed against five antibiotics such as amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. During this study, a total 40 streptococcal isolates were obtained out of which 2 isolates were of S. equi subsp. equi, 12 isolates were from S. equi subsp. zooepidemicus. In the PCR-based detection, we revealed amplicons of 235 bp and 679 bp for confirmation of sodA and seM gene, respectively. In antibiogram, two isolates of S. equi subsp. equi were found resistant to penicillin G, and all other isolates were found sensitive to amoxicillin and streptomycin. The majority of streptococcal infections was due to S. equi subsp. Zooepidemicus, and thus was recognized as a potential pathogen of diseases of equines besides S. equi subsp. equi.

  16. Surveillance of Mycobacterium avium subsp. paratuberculosis in dairy herds

    NARCIS (Netherlands)

    Weber, M.F.

    2009-01-01

    In this thesis, the potential for improvements in surveillance of Mycobacterium avium subsp. paratuberculosis (Map) infection and paratuberculosis in dairy herds was investigated, leading to a reduction in surveillance costs whilst continuing to meet specific quality targets. In particular,

  17. Knowledge on Sclerocarya birrea subsp. caffra with emphasis on its ...

    African Journals Online (AJOL)

    Knowledge on Sclerocarya birrea subsp. caffra with emphasis on its importance as a non-timber forest product in South and southern Africa: a summary: Part 1: Taxonomy, ecology and role in rural livelihoods: review paper.

  18. Description of Klebsiella quasipneumoniae sp. nov., isolated from human infections, with two subspecies, Klebsiella quasipneumoniae subsp. quasipneumoniae subsp. nov. and Klebsiella quasipneumoniae subsp. similipneumoniae subsp. nov., and demonstration that Klebsiella singaporensis is a junior heterotypic synonym of Klebsiella variicola.

    Science.gov (United States)

    Brisse, Sylvain; Passet, Virginie; Grimont, Patrick A D

    2014-09-01

    Strains previously classified as members of Klebsiella pneumoniae phylogroups KpI, KpII-A, KpII-B and KpIII were characterized by 16S rRNA (rrs) gene sequencing, multilocus sequence analysis based on rpoB, fusA, gapA, gyrA and leuS genes, average nucleotide identity and biochemical characteristics. Phylogenetic analysis demonstrated that KpI and KpIII corresponded to K. pneumoniae and Klebsiella variicola, respectively, whereas KpII-A and KpII-B formed two well-demarcated sequence clusters distinct from other members of the genus Klebsiella. Average nucleotide identity between KpII-A and KpII-B was 96.4 %, whereas values lower than 94 % were obtained for both groups when compared with K. pneumoniae and K. variicola. Biochemical properties differentiated KpII-A, KpII-B, K. pneumoniae and K. variicola, with acid production from adonitol and l-sorbose and ability to use 3-phenylproprionate, 5-keto-d-gluconate and tricarballylic acid as sole carbon sources being particularly useful. Based on their genetic and phenotypic characteristics, we propose the names Klebsiella quasipneumoniae subsp. quasipneumoniae subsp. nov. and K. quasipneumoniae subsp. similipneumoniae subsp. nov. for strains of KpII-A and KpII-B, respectively. The type strain of K. quasipneumoniae sp. nov. and of K. quasipneumoniae subsp. quasipneumoniae subsp. nov. is 01A030(T) ( = SB11(T) = CIP 110771(T) = DSM 28211(T)). The type strain of K. quasipneumoniae subsp. similipneumoniae subsp. nov. is 07A044(T) ( = SB30(T) = CIP 110770(T) = DSM 28212(T)). Both strains were isolated from human blood cultures. This work also showed that Klebsiella singaporensis is a junior heterotypic synonym of K. variicola. © 2014 IUMS.

  19. Diversity of the subspecies Bifidobacterium animalis subsp. lactis.

    Science.gov (United States)

    Bunesova, Vera; Killer, Jiri; Javurkova, Barbora; Vlkova, Eva; Tejnecky, Vaclav; Musilova, Sarka; Rada, Vojtech

    2017-04-01

    Strains of Bifidobacterium animalis subsp. lactis are well-known health-promoting probiotics used commercially. B. animalis subsp. lactis has been isolated from different sources, and little is known about animal isolates of this taxon. The aim of this study was to examine the genotypic and phenotypic diversity between B. animalis subsp. lactis strains different animal hosts including Cameroon sheep, Barbary sheep, okapi, mouflon, German shepard and to compare to BB12, food isolates and the collection strain DSM 10140. Ten strains of B. animalis subsp. lactis from different sources were characterised by phenotyping, fingerprinting, and multilocus sequence typing (MLST). Regardless of origin, MLST and phylogenetic analyses revealed a close relationship between strains of B. animalis subsp. lactis with commercial and animal origin with the exception of isolates from ovine cheese, mouflon and German Shepard dog. Moreover, isolates from dog and mouflon showed significant differences in fermentation profiles and peptide mass fingerprints (MALDI-TOF). Results indicated phenotypic and genotypic diversity among strains of B. animalis subsp. lactis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Susceptibilidade antimicrobiana de Campylobacter fetus subsp. venerealis isolado de bovinos Antimicrobial susceptibility of Campylobacter fetus subsp. venerealis isolated from cattle

    Directory of Open Access Journals (Sweden)

    Agueda C. Vargas

    2005-03-01

    Full Text Available A campilobacteriose venérea bovina, ocasionada principalmente pelo Campylobacter fetus subsp. fetus e Campylobacter subsp. venerealis, é transmitida através do coito ou por inseminação com sêmen contaminado. O propósito deste estudo foi determinar a susceptibilidade in vitro de isolados de C. fetus subesp. venerealis a agentes antimicrobianos comumente utilizados para o tratamento clínico e de sêmen. Foram testadas duas cepas padrão, sendo uma de C. fetus subsp. fetus e outra de C. fetus subsp. venerealis, bem como 21 amostras de isolados clínicos de C. fetus subsp. venerealis. Os testes foram realizados conforme o método de Kirby-Bauer. A amostra padrão de C. fetus subsp. fetus demonstrou-se resistente à lincomicina, penicilina e ácido nalidíxico, enquanto a de C. fetus subsp. venerealis apresentou susceptibilidade a todos antimicrobianos testados, com exceção do ácido nalidíxico. Todas as amostras de C. fetus subsp. venerealis foram susceptíveis à amicacina, ampicilina, cefalotina, estreptomicina, gentamicina, penicilina e tetraciclina. Foi observada resistência de 42,86% à lincomicina e 4,76 % a enrofloxacina, e de 100% ao ácido nalidíxico. Ainda, 4,76% apresentaram susceptibilidade intermediária à enrofloxacina, neomicina e polimixina B e 9,52% à lincomicina. Os resultados evidenciaram a sensibilidade das amostras analisadas aos antimicrobianos comumente utilizados para o tratamento clínico e do sêmen.Venereal campylobacteriosis is associated with infection of Campylobacter fetus subsp. fetus and Campylobacter fetus subsp. venerealis. The etiological agent is transmitted by natural bull breeding or artificial insemination using contaminated semen. The present study aimed to determine the in vitro susceptibility of C. fetus subsp. venerealis isolates to antimicrobial drugs generally used in clinical and semen treatment. Reference strains of C. fetus subsp. fetus and C. fetus subsp. venerealis and 21 C. fetus

  1. Prevalence of Streptococcus dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus in a sample of healthy dogs, cats and horses.

    Science.gov (United States)

    Acke, E; Midwinter, A C; Lawrence, K; Gordon, S J G; Moore, S; Rasiah, I; Steward, K; French, N; Waller, A

    2015-09-01

    To estimate the prevalence of β-haemolytic Lancefield group C streptococci in healthy dogs, cats and horses; to determine if frequent contact with horses was associated with isolation of these species from dogs and cats; and to characterise recovered S. equi subsp. zooepidemicus isolates by multilocus sequence typing. Oropharyngeal swabs were collected from 197 dogs and 72 cats, and nasopharyngeal swabs from 93 horses. Sampling was carried out at the Massey University Veterinary Teaching Hospital, on sheep and beef farms or on premises where horses were present. All animals were healthy and were categorised as Urban dogs and cats (minimal contact with horses or farm livestock), Farm dogs (minimal contact with horses) and Stable dogs and cats (frequent contact with horses). Swabs were cultured for β-haemolytic Streptococcus spp. and Lancefield group C streptococcal subspecies were confirmed by phenotypic and molecular techniques. Of the 197 dogs sampled, 21 (10.7 (95% CI= 4.0-25.4)%) tested positive for S. dysgalactiae subsp. equisimilis and 4 (2.0 (95% CI=0.7-5.5)%) tested positive for S. equi subsp. zooepidemicus. All these isolates, except for one S. dysgalactiae subsp. equisimilis isolate in an Urban dog, were from Stable dogs. S. dysgalactiae subsp. equisimilis was isolated from one Stable cat. Of the 93 horses, 22 (23.7 (95% CI=12.3-40.6)%) and 6 (6.5 (95% CI=2.8-14.1)%) had confirmed S. dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus isolation respectively. Isolation of S. dysgalactiae subsp. equisimilis from dogs was associated with frequent contact with horses (OR=9.8 (95% CI=2.6-72.8)). Three different multilocus sequence type profiles of S. equi subsp. zooepidemicus that have not been previously reported in dogs were recovered. Subclinical infection or colonisation by S. equi subsp. zooepidemicus and S. dysgalactiae subsp. equisimilis occurs in dogs and further research on inter-species transmission and the pathogenic potential of these

  2. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines

    OpenAIRE

    R. Javed; A. K. Taku; Rakhi Gangil; R. K. Sharma

    2016-01-01

    Aim: The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra), characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. Materials and Methods: A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease) and apparently healthy animals a...

  3. Biofilm formation of Francisella noatunensis subsp. orientalis

    Science.gov (United States)

    Soto, Esteban; Halliday-Wimmonds, Iona; Francis , Stewart; Kearney, Michael T.; Hansen, John D.

    2015-01-01

    Francisella noatunensis subsp. orientalis (Fno) is an emergent fish pathogen in both marine and fresh water environments. The bacterium is suspected to persist in the environment even without the presence of a suitable fish host. In the present study, the influence of different abiotic factors such as salinity and temperature were used to study the biofilm formation of different isolates of Fno including intracellular growth loci C (iglC)and pathogenicity determinant protein A (pdpA) knockout strains. Finally, we compared the susceptibility of planktonic and biofilm to three disinfectants used in the aquaculture and ornamental fish industry, namely Virkon®, bleach and hydrogen peroxide. The data indicates that Fno is capable of producing biofilms within 24 h where both salinity as well as temperature plays a role in the growth and biofilm formation of Fno. Mutations in theiglC or pdpA, both known virulence factors, do not appear to affect the capacity of Fno to produce biofilms, and the minimum inhibitory concentration, and minimum biocidal concentration for the three disinfectants were lower than the minimum biofilm eradication concentration values. This information needs to be taken into account if trying to eradicate the pathogen from aquaculture facilities or aquariums.

  4. New phenylethanoids from Buddleja cordata subsp. cordata.

    Science.gov (United States)

    Acevedo, L; Martínez, E; Castañeda, P; Franzblau, S; Timmermann, B N; Linares, E; Bye, R; Mata, R

    2000-04-01

    Bioassay-guided fractionation of a crude extract of the stem bark of Buddleja cordata subsp. cordata with significant antimycobacterial activity led to the isolation of a mixture composed by ten new long-chain esters of 2[4'-hydroxyphenyl]-ethanol (1-10), along with the lichen metabolites methyl beta-orcinolcarboxylate (11) and beta-orcinolcarboxylate (12). Extensive HPLC allowed the separation of the major components of the mixture, which were characterized by spectral means as 2[4'-hydroxyphenyl]-ethyl stearate (3), 2[4'-hydroxyphenyl]-ethyl behenate (6), and 2[4'-hydroxyphenyl]-ethyl lignocerate (8). The minor esters were identified as 2[4'-hydroxyphenyl]-ethyl palmitate (1), 2[4'-hydroxyphenyl]-ethyl heptadecanoate (2), 2[4'-hydroxyphenyl]-ethyl nonadecanoate (4), 2[4'-hydroxyphenyl]-ethyl arachidate (5), 2[4'-hydroxyphenyl]-ethyl tricosanoate (7), 2[4'-hydroxyphenyl]-ethyl pentacosanoate (9), and 2[4'-hydroxyphenyl]-ethyl hexacosanoate (10) by GC-MS analysis of the methyl esters derivatives of the fatty acids obtained by alkaline hydrolysis of the mixture. Compound 8 exhibited moderate antibacterial activity against Mycobacterium tuberculosis (MIC = 64 micrograms/ml).

  5. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines

    Science.gov (United States)

    Javed, R.; Taku, A. K.; Gangil, Rakhi; Sharma, R. K.

    2016-01-01

    Aim: The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra), characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. Materials and Methods: A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease) and apparently healthy animals and were sent to laboratory. The organisms were isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and confirmed by cultural characteristics and biochemical tests. Molecular detection of Streptococcus was done directly from cultures using sodA and seM gene-based polymerase chain reaction (PCR). Antibiogram was performed against five antibiotics such as amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During this study, a total 40 streptococcal isolates were obtained out of which 2 isolates were of S. equi subsp. equi, 12 isolates were from S. equi subsp. zooepidemicus. In the PCR-based detection, we revealed amplicons of 235 bp and 679 bp for confirmation of sodA and seM gene, respectively. In antibiogram, two isolates of S. equi subsp. equi were found resistant to penicillin G, and all other isolates were found sensitive to amoxicillin and streptomycin. Conclusion: The majority of streptococcal infections was due to S. equi subsp. Zooepidemicus, and thus was recognized as a potential pathogen of diseases of equines besides S. equi subsp. equi. PMID:27651677

  6. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines

    Directory of Open Access Journals (Sweden)

    R. Javed

    2016-08-01

    Full Text Available Aim: The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra, characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. Materials and Methods: A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease and apparently healthy animals and were sent to laboratory. The organisms were isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and confirmed by cultural characteristics and biochemical tests. Molecular detection of Streptococcus was done directly from cultures using sodA and seM gene-based polymerase chain reaction (PCR. Antibiogram was performed against five antibiotics such as amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During this study, a total 40 streptococcal isolates were obtained out of which 2 isolates were of S. equi subsp. equi, 12 isolates were from S. equi subsp. zooepidemicus. In the PCR-based detection, we revealed amplicons of 235 bp and 679 bp for confirmation of sodA and seM gene, respectively. In antibiogram, two isolates of S. equi subsp. equi were found resistant to penicillin G, and all other isolates were found sensitive to amoxicillin and streptomycin. Conclusion: The majority of streptococcal infections was due to S. equi subsp. Zooepidemicus, and thus was recognized as a potential pathogen of diseases of equines besides S. equi subsp. equi.

  7. Anoxybacillus kamchatkensis subsp. asaccharedens subsp. nov., a thermophilic bacterium isolated from a hot spring in Batman.

    Science.gov (United States)

    Gul-Guven, Reyhan; Guven, Kemal; Poli, Annarita; Nicolaus, Barbara

    2008-12-01

    A new thermophilic spore-forming strain KG8(T) was isolated from the mud of Taslidere hot spring in Batman. Strain KG8(T) was aerobe, Gram-positive, rod-shaped, motile, occurring in pairs or filamentous. Growth was observed from 35-65 degrees C (optimum 55 degrees C) and at pH 5.5-9.5 (optimum pH 7.5). It was capable of utilizing starch, growth was observed until 3% NaCl (w/v) and it was positive for nitrate reduction. On the basis of 16S rRNA gene sequence similarity, strain KG8(T) was shown to be related most closely to Anoxybacillus species. Chemotaxonomic data (major isoprenoid quinone-menaquinone-7; major fatty acid-iso-C15:0 and iso-C17:0) supported the affiliation of strain KG8(T) to the genus Anoxybacillus. The results of DNA-DNA hybridization, physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain KG8(T). Based on these results we propose assigning a novel subspecies of Anoxybacillus kamchatkensis, to be named Anoxybacillus kamchatkensis subsp. asaccharedens subsp. nov. with the type strain KG8(T) (DSM 18475(T)=CIP 109280(T)).

  8. Common genomic features of Campylobacter jejuni subsp. doylei strains distinguish them from C. jejuni subsp. jejuni

    Directory of Open Access Journals (Sweden)

    Horn Sharon T

    2007-05-01

    Full Text Available Abstract Background Campylobacter jejuni has been divided into two subspecies: C. jejuni subsp. jejuni (Cjj and C. jejuni subsp. doylei (Cjd. Nearly all of the C. jejuni strains isolated are Cjj; nevertheless, although Cjd strains are isolated infrequently, they differ from Cjj in two key aspects: they are obtained primarily from human clinical samples and are associated often with bacteremia, in addition to gastroenteritis. In this study, we utilized multilocus sequence typing (MLST and a DNA microarray-based comparative genomic indexing (CGI approach to examine the genomic diversity and gene content of Cjd strains. Results A geographically diverse collection of eight Cjd strains was examined by MLST and determined to be phylogenetically distinct from Cjj strains. Microarray-based CGI approach also supported this. We were able to demonstrate that Cjd strains exhibited divergence from Cjj strains NCTC 11168 and RM1221 in many of the intraspecies hypervariable regions. Moreover, multiple metabolic, transport and virulence functions (e.g. cytolethal distending toxin were shown to be absent in the Cjd strains examined. Conclusion Our data demonstrate that Cjd are phylogenetically distinct from Cjj strains. Using the CGI approach, we identified subsets of absent genes from amongst the C. jejuni genes that provide clues as to the potential evolutionary origin and unusual pathogenicity of Cjd.

  9. Potential Transmission Pathways of Streptococcus gallolyticus subsp. gallolyticus.

    Directory of Open Access Journals (Sweden)

    Jessika Dumke

    Full Text Available Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus subsp. gallolyticus, a member of group D streptococci, is an inhabitant of the animal and human gastrointestinal tract. Furthermore, it is a facultative pathogen which causes e.g. endocarditis, septicemia and mastitis. S. gallolyticus subsp. gallolyticus may be transmitted either directly or indirectly between animals and humans. However, the transmission routes are an unsolved issue. In this study, we present systematic analyses of an S. gallolyticus subsp. gallolyticus isolate of an infective endocarditis patient in relation to isolates of his laying hen flock. Isolates from pooled droppings of laying hens, pooled dust samples and human blood culture were characterized by using multilocus sequence typing (MLST and DNA fingerprinting. MLST revealed the same allelic profile of isolates from the human blood culture and from the droppings of laying hens. In addition, these isolates showed clonal identity regarding a similar DNA fingerprinting pattern. For the first time, we received a hint that transmission of S. gallolyticus subsp. gallolyticus between poultry and humans may occur. This raises the question about the zoonotic potential of isolates from poultry and should be considered in future studies.

  10. A single or multistage mycobacterium avium subsp. paratuberculosis subunit vaccine

    DEFF Research Database (Denmark)

    2014-01-01

    The present invention provides one or more immunogenic polypeptides for use in a preventive or therapeutic vaccine against latent or active infection in a human or animal caused by a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis. Furthermore a single or multi-phase vaccine...... comprising the one or more immunogenic polypeptides is provided for administration for the prevention or treatment of infection with a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis. Additionally, nucleic acid vaccines, capable of in vivo expression of the multi-phase vaccine...... comprising the one or more immunogenic polypeptides, is provided for prevention or treatment of infection with a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis....

  11. Campylobacter hyointestinalis subsp hyointestinalis, a common Campylobacter species in reindeer

    DEFF Research Database (Denmark)

    Hanninen, M.L.; Sarelli, L.; Sukura, A.

    2002-01-01

    Aims: To study the prevalence of Campylobacter spp. in the faecal material of reindeer, and to identify the isolates by means of a polyphasic approach. In addition, to study the genetic diversity of Camp. hyointestinalis subsp. hyointestinalis reindeer isolates by pulsed-field gel electrophoresis...... slaughterhouses. Samples were cultured by methods suitable for isolation of fastidious Campylobacter species. Of all samples, 6% (24/399) were Campylobacter-positive. Phenotypic characteristics, SDS-PAGE protein patterns, dot blot DNA-DNA hybridization, 23S rDNA restriction fragment polymorphism analysis and PFGE...... identified the isolates as Camp. hyointestinalis subsp. kyointestinalis. Conclusions: Campylobacter hyointestinalis subsp. hyointestinalis was the only Campylobacter species isolated from reindeer in this study. The isolates showed high genomic diversity in PFGE with the restriction enzymes SmaI and Kpn...

  12. A single or multistage mycobacterium avium subsp. paratuberculosis subunit vaccine

    DEFF Research Database (Denmark)

    2014-01-01

    The present invention provides one or more immunogenic polypeptides for use in a preventive or therapeutic vaccine against latent or active infection in a human or animal caused by a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis. Furthermore a single or multi-phase vaccine...... comprising the one or more immunogenic polypeptides is provided for administration for the prevention or treatment of infection with a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis. Additionally, nucleic acid vaccines, capable of in vivo expression of the multi-phase vaccine...

  13. Vitaliana primuliflora subsp. assoana. Nueva cita para la Comunidad Valenciana (Vitaliana primuliflora subsp. assoana, new record for valencian flora

    Directory of Open Access Journals (Sweden)

    Amparo OLIVARES

    2012-04-01

    Full Text Available Se comunica el hallazgo de una localidad situada entre el límite noroeste de la provincia de Valencia y el sureste de la de Teruel donde se ha encontrado Vitaliana primuliflora subsp. assoana, nueva población de esta especie para la Comunidad Valenciana.SUMMARY: This paper reports the discovery of a new location from the northwest boundary of the province of Valencia and south of Teruel (E Spain where found Vitaliana primuliflora subsp. assoana, new population of this species to Valencia.

  14. Fusobacterium nucleatum subsp. fusiforme Gharbia and Shah 1992 is a later synonym of Fusobacterium nucleatum subsp. vincentii Dzink et al. 1990.

    Science.gov (United States)

    Kook, Joong-Ki; Park, Soon-Nang; Lim, Yun Kyong; Choi, Mi-Hwa; Cho, Eugene; Kong, Si-Won; Shin, Yeseul; Paek, Jayoung; Chang, Young-Hyo

    2013-04-01

    On the basis of the DNA-DNA hybridization patterns and phenotypic characteristics, Fusobacterium nucleatum was classified into five subspecies. Previous studies have suggested that F. nucleatum subsp. vincentii is genetically similar to F. nucleatum subsp. fusiforme. The aim of this study was to investigate the possibility of classifying these two subspecies into a single subspecies by phylogenetic analysis using a single sequence (24,715 bp) concatenated 22 housekeeping genes of eight F. nucleatum strains including type strains of five F. nucleatum subspecies. The phylogenetic analysis indicated that F. nucleatum subsp. vincentii and F. nucleatum subsp. fusiforme were clustered in the same group and each strain of other F. nucleatum subspecies were also separated into the same cluster. These results suggested that F. nucleatum subsp. fusiforme and F. nucleatum subsp. vincentii can be classified into a single subspecies. F. nucleatum subsp. vincentii was early published name; therefore, F. nucleatum subsp. fusiforme Gharbia and Shah 1992 can be regarded as a later synonym of F. nucleatum subsp. vincentii Dzink et al. 1990.

  15. Laminaria japonica Extract, an Inhibitor of Clavibater michiganense Subsp. Sepedonicum.

    Directory of Open Access Journals (Sweden)

    Jin Cai

    Full Text Available Bacterial ring rot of potato is one of the most serious potato plant and tuber diseases. Laminaria japonica extract was investigated for its antimicrobial activity against Clavibater michiganense subsp. sepedonicum (Spieckermann & Kotthoff Davis et al., the causative agent of bacterial ring rot of potato. The results showed that the optimum extraction conditions of antimicrobial substances from L. japonica were an extraction temperature of 80°C, an extraction time of 12 h, and a solid to liquid ratio of 1∶25. Active compounds of L. japonica were isolated by solvent partition, thin layer chromatography (TLC and column chromatography. All nineteen fractionations had antimicrobial activities against C. michiganense subsp. sepedonicum, while Fractionation three (Fr.3 had the highest (P<0.05 antimicrobial activity. Chemical composition analysis identified a total of 26 components in Fr.3. The main constituents of Fr.3 were alkanes (80.97%, esters (5.24%, acids (4.87% and alcohols (2.21%. Antimicrobial activity of Fr.3 against C. michiganense subsp. sepedonicum could be attributed to its ability to damage the cell wall and cell membrane, induce the production of reactive oxygen species (ROS, increase cytosolic Ca2+ concentration, inhibit the glycolytic pathway (EMP and tricarboxylic acid (TCA cycle, inhibit protein and nucleic acid synthesis, and disrupt the normal cycle of DNA replication. These findings indicate that L. japonica extracts have potential for inhibiting C. michiganense subsp. sepedonicum.

  16. Laminaria japonica Extract, an Inhibitor of Clavibater michiganense Subsp. Sepedonicum.

    Science.gov (United States)

    Cai, Jin; Feng, Jia; Xie, Shulian; Wang, Feipeng; Xu, Qiufeng

    2014-01-01

    Bacterial ring rot of potato is one of the most serious potato plant and tuber diseases. Laminaria japonica extract was investigated for its antimicrobial activity against Clavibater michiganense subsp. sepedonicum (Spieckermann & Kotthoff) Davis et al., the causative agent of bacterial ring rot of potato. The results showed that the optimum extraction conditions of antimicrobial substances from L. japonica were an extraction temperature of 80°C, an extraction time of 12 h, and a solid to liquid ratio of 1∶25. Active compounds of L. japonica were isolated by solvent partition, thin layer chromatography (TLC) and column chromatography. All nineteen fractionations had antimicrobial activities against C. michiganense subsp. sepedonicum, while Fractionation three (Fr.3) had the highest (P<0.05) antimicrobial activity. Chemical composition analysis identified a total of 26 components in Fr.3. The main constituents of Fr.3 were alkanes (80.97%), esters (5.24%), acids (4.87%) and alcohols (2.21%). Antimicrobial activity of Fr.3 against C. michiganense subsp. sepedonicum could be attributed to its ability to damage the cell wall and cell membrane, induce the production of reactive oxygen species (ROS), increase cytosolic Ca2+ concentration, inhibit the glycolytic pathway (EMP) and tricarboxylic acid (TCA) cycle, inhibit protein and nucleic acid synthesis, and disrupt the normal cycle of DNA replication. These findings indicate that L. japonica extracts have potential for inhibiting C. michiganense subsp. sepedonicum.

  17. Mycobacterium avium subsp. paratuberculosis infection, immunology and pathology of livestock

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) infection in ruminants leads to a chronic and progressive enteric disease (Johne’s disease) that results in loss of intestinal function, poor body condition, and eventual death. Transmission is primarily through a fecal-oral route in neonates but con...

  18. Comparative genomics of extrachromosomal elements in Bacillus thuringiensis subsp. israelensis.

    Science.gov (United States)

    Bolotin, Alexandre; Gillis, Annika; Sanchis, Vincent; Nielsen-LeRoux, Christina; Mahillon, Jacques; Lereclus, Didier; Sorokin, Alexei

    2017-05-01

    Bacillus thuringiensis subsp. israelensis is one of the most important microorganisms used against mosquitoes. It was intensively studied following its discovery and became a model bacterium of the B. thuringiensis species. Those studies focused on toxin genes, aggregation-associated conjugation, linear genome phages, etc. Recent announcements of genomic sequences of different strains have not been explicitly related to the biological properties studied. We report data on plasmid content analysis of four strains using ultra-high-throughput sequencing. The strains were commercial product isolates, with their putative ancestor and type B. thuringiensis subsp. israelensis strain sequenced earlier. The assembled contigs corresponding to published and novel data were assigned to plasmids described earlier in B. thuringiensis subsp. israelensis and other B. thuringiensis strains. A new 360 kb plasmid was identified, encoding multiple transporters, also found in most of the earlier sequenced strains. Our genomic data show the presence of two toxin-coding plasmids of 128 and 100 kb instead of the reported 225 kb plasmid, a co-integrate of the former two. In two of the sequenced strains, only a 100 kb plasmid was present. Some heterogeneity exists in the small plasmid content and structure between strains. These data support the perception of active plasmid exchange among B. thuringiensis subsp. israelensis strains in nature. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  19. Molecular Characterization of Bartonella vinsonii subsp. berkhoffii Genotype III▿

    Science.gov (United States)

    Cadenas, Maria B.; Bradley, Julie; Maggi, Ricardo G.; Takara, Matt; Hegarty, Barbara C.; Breitschwerdt, Edward B.

    2008-01-01

    The molecular characterization of a Bartonella vinsonii subsp. berkhoffii genotype III strain (NCSU strain 06-CO1) isolated from the blood of a military working dog diagnosed with endocarditis is reported in this study. Several genes were amplified and sequenced for comparative sequence similarity with other strains. PMID:18367567

  20. New Iridoid Glycosides from Lamium eriocephalum subsp. eriocephalum

    DEFF Research Database (Denmark)

    Yalcin, Funda Nuray; Ersöz, Tayfun; Avci, Kürsat

    2007-01-01

    Two new iridoid glycosides, eriobioside (1) and lamerioside (2) were isolated from the aerial parts of Lamium eriocephalum subsp. eriocephalum, along with two known componds, lamiide (3) and ipolamiide (4). Their structures were elucidated by spectroscopic methods (UV, 1D- and 2D-NMR) and by mass...

  1. Fitness and its variation among populations of Acacia tortilis subsp ...

    African Journals Online (AJOL)

    Administrator

    2008-07-23

    Jul 23, 2008 ... Acacia tortilis subsp. raddiana (Fabaceae) plays an important role in the life of desert animals and is a major source of livestock feed ... raddiana grows in desert wadis and sandy plains, usually in water catchments areas in ...... Randomization and Monte Carlo Methods in Biology. Chapman & Hall, London.

  2. Streptococcus equi subsp. zooepidemicus Infections Associated with Guinea Pigs

    Science.gov (United States)

    Young, Andrea; Levine, Seth J.; Garvin, Joseph P.; Brown, Susan; Turner, Lauren; Fritzinger, Angela; Gertz, Robert E.; Murphy, Julia M.; Vogt, Marshall; Beall, Bernard

    2015-01-01

    Streptococcus equi subsp. zooepidemicus is a known zoonotic pathogen. In this public health investigation conducted in Virginia, USA, in 2013, we identified a probable family cluster of S. zooepidemicus cases linked epidemiologically and genetically to infected guinea pigs. S. zooepidemicus infections should be considered in patients who have severe clinical illness and report guinea pig exposure. PMID:25531424

  3. Recurrent Streptococcus equi subsp. zooepidemicus Bacteremia in an Infant.

    Science.gov (United States)

    Watson, Joshua R; Leber, Amy; Velineni, Sridhar; Timoney, John F; Ardura, Monica I

    2015-09-01

    We describe a case of an infant with recurrent bacteremia caused by Streptococcus equi subsp. zooepidemicus, likely transmitted from mother to infant. Our case highlights the importance of an epidemiological history and molecular diagnostics in ascertaining insights into transmission, pathogenesis, and optimal management. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. Complete Genome Sequence of Beijerinckia indica subsp. indica▿

    Science.gov (United States)

    Tamas, Ivica; Dedysh, Svetlana N.; Liesack, Werner; Stott, Matthew B.; Alam, Maqsudul; Murrell, J. Colin; Dunfield, Peter F.

    2010-01-01

    Beijerinckia indica subsp. indica is an aerobic, acidophilic, exopolysaccharide-producing, N2-fixing soil bacterium. It is a generalist chemoorganotroph that is phylogenetically closely related to facultative and obligate methanotrophs of the genera Methylocella and Methylocapsa. Here we report the full genome sequence of this bacterium. PMID:20601475

  5. Complete genome sequence of Beijerinckia indica subsp. indica.

    Science.gov (United States)

    Tamas, Ivica; Dedysh, Svetlana N; Liesack, Werner; Stott, Matthew B; Alam, Maqsudul; Murrell, J Colin; Dunfield, Peter F

    2010-09-01

    Beijerinckia indica subsp. indica is an aerobic, acidophilic, exopolysaccharide-producing, N(2)-fixing soil bacterium. It is a generalist chemoorganotroph that is phylogenetically closely related to facultative and obligate methanotrophs of the genera Methylocella and Methylocapsa. Here we report the full genome sequence of this bacterium.

  6. Factors affecting survival of Clavibacter michiganesis subsp. sepedonicus in water

    NARCIS (Netherlands)

    Wolf, van der J.M.; Beckhoven, van J.R.C.M.

    2004-01-01

    The survival of Clavibacter michiganensis subsp. sepedonicus (Cms), the causal organism of bacterial ring rot in potato, was studied in water, to assess the risks for dissemination of Cms via surface water and infection of potato crops by irrigation. Cms was able to survive for a maximum period of 7

  7. The effects of aqueous root extract of Senna italica subsp ...

    African Journals Online (AJOL)

    The effects of aqueous root extracts of Senna italica subsp. arachoides on the feeding performance of adults of Hyalomma marginatum rufipes in three consecutive infestations of rabbits were studied under laboratory conditions. Rabbits were divided into treatment group (n = 3), fed aqueous root extracts and rabbit pellets ...

  8. Knowledge on Sclerocarya birrea subsp. caffra with emphasis on its ...

    African Journals Online (AJOL)

    Knowledge on Sclerocarya birrea subsp. caffra with emphasis on its importance as a non-timber forest product in South and southern Africa: a summary: part 2: commercial use, tenure and policy, domestication, intellectual property rights and benefit-sharing: review paper.

  9. Plant regeneration of Brassica oleracea subsp. italica (Broccoli) CV ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor Darul. Ehsan, Malaysia. Accepted 20 March, 2009. Hypocotyls and shoot tips were used as explants in in vitro plant regeneration of broccoli (Brassica oleracea subsp.italica) cv. Green Marvel.

  10. Fitness and its variation among populations of Acacia tortilis subsp ...

    African Journals Online (AJOL)

    Therefore, this study aims to determine if A. tortilis subsp. raddiana populations suffer reduced fitness and its correlation or association with genetic diversity and mating parameters. Correlations and association between fitness, population size, genetic variation, and mating system parameters were tested using Spearman ...

  11. Sensitive and rapid detection of Mycoplasma capricolum subsp ...

    African Journals Online (AJOL)

    SAM

    2014-05-21

    May 21, 2014 ... The LAMP assay was able to detect MCCP in tissue. Key words: Mycoplasma capricolum subsp. Capripneumoniae, loop-mediated isothermal amplification, rapid detection. INTRODUCTION. Contagious caprine pleuropneumonia (CCPP) is a severe infectious disease of goats caused by Mycoplasma.

  12. Novel Temperate Phages of Salmonella enterica subsp. salamae and subsp. diarizonae and Their Activity against Pathogenic S. enterica subsp. enterica Isolates.

    Science.gov (United States)

    Mikalová, Lenka; Bosák, Juraj; Hříbková, Hana; Dědičová, Daniela; Benada, Oldřich; Šmarda, Jan; Šmajs, David

    2017-01-01

    Forty strains of Salmonella enterica (S. enterica) subspecies salamae (II), arizonae (IIIa), diarizonae (IIIb), and houtenae (IV) were isolated from human or environmental samples and tested for bacteriophage production. Production of bacteriophages was observed in 15 S. enterica strains (37.5%) belonging to either the subspecies salamae (8 strains) or diarizonae (7 strains). Activity of phages was tested against 52 pathogenic S. enterica subsp. enterica isolates and showed that phages produced by subsp. salamae had broader activity against pathogenic salmonellae compared to phages from the subsp. diarizonae. All 15 phages were analyzed using PCR amplification of phage-specific regions and 9 different amplification profiles were identified. Five phages (SEN1, SEN4, SEN5, SEN22, and SEN34) were completely sequenced and classified as temperate phages. Phages SEN4 and SEN5 were genetically identical, thus representing a single phage type (i.e. SEN4/5). SEN1 and SEN4/5 fit into the group of P2-like phages, while the SEN22 phage showed sequence relatedness to P22-like phages. Interestingly, while phage SEN34 was genetically distantly related to Lambda-like phages (Siphoviridae), it had the morphology of the Myoviridae family. Based on sequence analysis and electron microscopy, phages SEN1 and SEN4/5 were members of the Myoviridae family and phage SEN22 belonged to the Podoviridae family.

  13. Molecular identification of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in nasal swabs samples from horses suffering respiratory infections in Iran.

    Science.gov (United States)

    Jannatabadi, A A; Mohammadi, G R; Rad, M; Maleki, M

    2008-02-01

    The objective of this study was to evaluate the existence of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus as probable agents associated with naturally occurring infection of the equine upper respiratory disease in Mashhad area. Nasal swabs samples from thirty horses with upper respiratory tract infections were collected. The bacteria isolated and identified were Streptococcus equi subsp. equi (1 isolate), Streptococcus equi subsp. zooepidemicus (25 isolates), Pasteurella sp. (11 isolates), Staphylococcus sp. (17 isolates), Bacillus sp. (4 isolates), Pseudomonas sp. (4 isolates), Proteus sp. (1 isolate), Neisseria sp. (1 isolate) and E. coli (1 isolate). All 25 isolates of Streptococcus equi subsp. zooepidemicus and the isolate of Streptococcus equi subsp. equi were characterized by biochemical tests and molecular techniques. For molecular identification of the subspecies S. equi and S. zooepidemicus two genomic region SeM and sodA were amplified. This study is the first report of molecular identification of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in Iran.

  14. Dose-response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults

    DEFF Research Database (Denmark)

    Larsen, C.N.; Nielsen, S.; Kaestel, P.

    2006-01-01

    Objective: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora was analy...

  15. Actinobacillus equuli subsp. equuli associated with equine valvular endocarditis

    DEFF Research Database (Denmark)

    Aalbæk, Bent; Østergaard, Stine; Buhl, Rikke

    2007-01-01

    Microbiological and pathological data from a case of equine valvular endocarditis are reported. Limited information is available on the pathogenic potential of equine Actinobacillus species as several strains originate from apparently healthy horses. After the establishment of two subspecies within...... this species, this seems to be the first report of an etiological association between A. equuli subsp. equuli and equine endocarditis. Furthermore, new information on some phenotypical characteristics of this subspecies are reported, compared to previous findings...

  16. Russian isolates enlarge the known geographic diversity of Francisella tularensis subsp. mediasiatica.

    Directory of Open Access Journals (Sweden)

    Vitalii Timofeev

    Full Text Available Francisella tularensis, a small Gram-negative bacterium, is capable of infecting a wide range of animals, including humans, and causes a plague-like disease called tularemia-a highly contagious disease with a high mortality rate. Because of these characteristics, F. tularensis is considered a potential agent of biological terrorism. Currently, F. tularensis is divided into four subspecies, which differ in their virulence and geographic distribution. Two of them, subsp. tularensis (primarily found in North America and subsp. holarctica (widespread across the Northern Hemisphere, are responsible for tularemia in humans. Subsp. novicida is almost avirulent in humans. The fourth subspecies, subsp. mediasiatica, is the least studied because of its limited distribution and impact in human health. It is found only in sparsely populated regions of Central Asia. In this report, we describe the first focus of naturally circulating F. tularensis subsp. mediasiatica in Russia. We isolated and characterized 18 strains of this subspecies in the Altai region. All strains were highly virulent in mice. The virulence of subsp. mediasiatica in a vaccinated mouse model is intermediate between that of subsp. tularensis and subsp. holarctica. Based on a multiple-locus variable number tandem repeat analysis (MLVA, we show that the Altaic population of F. tularensis subsp. mediasiatica is genetically distinct from the classical Central Asian population, and probably is endemic to Southern Siberia. We propose to subdivide the mediasiatica subspecies into three phylogeographic groups, M.I, M.II and M.III.

  17. Colonization patterns of an mCherry-tagged Pectobacterium carotovorum subsp. brasiliense strain in potato plants.

    Science.gov (United States)

    Kubheka, Gugulethu C; Coutinho, Teresa A; Moleleki, Ntsane; Moleleki, Lucy N

    2013-12-01

    Pectobacterium carotovorum subsp. brasiliense is a newly identified member of the potato soft rot enterobacteriaceae. The pathogenesis of this pathogen is still poorly understood. In this study, an mCherry-P. carotovorum subsp. brasiliense-tagged strain was generated to study P. carotovorum subsp. brasiliense-potato plant interactions. Prior to use, the tagged strain was evaluated for in vitro growth, plasmid stability, and virulence on potato tubers and shown to be similar to the wild type. Four potato cultivars were evaluated for stem-based resistance against P. carotovorum subsp. brasiliense. Confocal laser-scanning microscopy and in vitro viable cell counts showed that P. carotovorum subsp. brasiliense is able to penetrate roots of a susceptible potato cultivar as early as 12 h postinoculation and migrate upward into aerial stem parts. Due to the phenotypic differences observed between tolerant and susceptible cultivars, a comparison of P. carotovorum subsp. brasiliense colonization patterns in these cultivars was undertaken. In the susceptible cultivar, P. carotovorum subsp. brasiliense cells colonized the xylem tissue, forming "biofilm-like" aggregates that led to occlusion of some of the vessels. In contrast, in the tolerant cultivar, P. carotovorum subsp. brasiliense appeared as free-swimming planktonic cells with no specific tissue localization. This suggests that there are resistance mechanisms in the tolerant cultivar that limit aggregation of P. carotovorum subsp. brasiliense in planta and, hence, the lack of symptom development in this cultivar.

  18. Bacillus amyloliquefaciens SUBSP. plantarum PROBIOTIC STRAINS AS PROTEASE PRODUCERS

    Directory of Open Access Journals (Sweden)

    E. V. Маtseliukh

    2015-04-01

    Full Text Available Proteases from probiotic strains of the genus Bacillus, just like the antibiotics, bacteriocins and other hydrolytic enzymes, are one of the main factors that determine their biological activity. The aim of this work was to study the synthesis and biochemical properties of proteases from two strains Bacillus amyloliquefaciens subsp. plantarum UCM B-5139 and UCM B-5140 that included in the probiotic Endosporin. The cultivation of strains was carried out in flasks under rotating for two days. The influence of physico-chemical parameters of the reaction medium on proteolytic activity was studied on partially purified protease preparations. Lytic activity was determined by turbidimetric method. On the second day of cultivation B. amyloliquefaciens subsp. plantarum UCM В-5139 and UCM В-5140 synthesized the metal-dependent peptidase and serine protease, respectively. The optimum conditions of their action were the following: temperature 37–40 °C and pH 6.5–7.0. Isolated proteases are able to lyse the living cells of Staphylococcus aureus and Candida albicans. Thus we demonstrated that B. amyloliquefaciens subsp. plantarum UCM B-5140 and UCM B-5139, included in the probiotic veterinary preparation Endosporin, produced proteolytic enzymes that hydrolyze the native insoluble proteins (elastin, fibrin and collagen. These enzymes belong to the group of neutral metal-dependent and serine proteases. They are active under physiological conditions against gram-positive bacteria and yeasts. The application of these proteases in biotechnology is considered.

  19. Discrimination of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

    Science.gov (United States)

    Mani, Rinosh J; Thachil, Anil J; Ramachandran, Akhilesh

    2017-09-01

    Accurate and timely identification of infectious etiologies is of great significance in veterinary microbiology, especially for critical diseases such as strangles, a highly contagious disease of horses caused by Streptococcus equi subsp. equi. We evaluated a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) platform for use in species- and subspecies-level identification of S. equi isolates from horses and compared it with an automated biochemical system. We used 25 clinical isolates each of S. equi subsp. equi and S. equi subsp. zooepidemicus. Using the MALDI-TOF MS platform, it was possible to correctly identify all 50 isolates to the species level. Unique mass peaks were identified in the bacterial peptide mass spectra generated by MALDI-TOF MS, which can be used for accurate subspecies-level identification of S. equi. Mass peaks (mass/charge, m/ z) 6,751.9 ± 1.4 (mean ± standard deviation) and 5,958.1 ± 1.3 were found to be unique to S. equi subsp. equi and S. equi subsp. zooepidemicus, respectively. The automated biochemical system correctly identified 47 of 50 of the isolates to the species level as S. equi, whereas at the subspecies level, 24 of 25 S. equi subsp. equi isolates and 22 of 25 S. equi subsp. zooepidemicus isolates were correctly identified. Our results indicate that MALDI-TOF MS can be used for accurate species- and subspecies-level identification of S. equi.

  20. Development of strain-specific PCR primers for the identification of Fusobacterium nucleatum subsp. fusiforme ATCC 51190(T) and subsp. vincentii ATCC 49256(T).

    Science.gov (United States)

    Shin, Hwan Seon; Kim, Min-Jung; Kim, Hwa-Sook; Park, Soon-Nang; Kim, Do Kyung; Baek, Dong-Heon; Kim, Chan; Kook, Joong-Ki

    2010-02-01

    The objective of this study was to develop the strain-specific PCR primers for Fusobacterium nucleatum subsp. fusiforme ATCC 51190(T) and F. nucleatum subsp. vincentii ATCC 49256(T) based on the nucleotide sequence of the Fs17 and Fv35 DNA probes, respectively. The strain specificity was tested against 10 type strains of Fusobacterium spp. or subsp., 21 clinical isolates of F. nucleatum from Koreans, and five type strains of distinct Fusobacterium species. Primer sensitivity was determined by testing serial dilutions (4 ng-4 fg) of the purified genomic DNA from each of the type strains. PCR showed that two pairs of PCR primers, Fs17-F14/Fs17-R14 and Fv35-F1/Fv35-R1 primers, could produce strain-specific amplicons from F. nucleatum subsp. fusiforme ATCC 51190(T) and F. nucleatum subsp. vincentii ATCC 49256(T), respectively. The two PCR primer sets could detect as little as 0.4 pg or 4 pg of the genomic DNA of each target strain. These results suggest that the two sets of PCR primers could be used to identify F. nucleatum subsp. fusiforme ATCC 51190(T) and F. nucleatum subsp. vincentii ATCC 49256(T), particularly for ascertaining the authenticity of the strain. 2009 Elsevier Ltd. All rights reserved.

  1. Stability evaluation of freeze-dried Lactobacillus paracasei subsp. tolerance and Lactobacillus delbrueckii subsp. bulgaricus in oral capsules

    OpenAIRE

    Jalali, M.; Abedi, D.; Varshosaz, J.; Najjarzadeh, M.; Mirlohi, M.; Tavakoli, N.

    2012-01-01

    Freeze-drying is a common preservation technology in the pharmaceutical industry. Various studies have investigated the effect of different cryoprotectants on probiotics during freeze-drying. However, information on the effect of cryoprotectants on the stability of some Lactobacillus strains during freeze-drying seems scarce. Therefore, the aim of the present study was to establish production methods for preparation of oral capsule probiotics containing Lactobacillus paracasei subsp. toleranc...

  2. Cloning, Sequencing, and Expression of the Pyruvate Carboxylase Gene in Lactococcus lactis subsp. lactis C2†

    OpenAIRE

    Wang, H.; O'Sullivan, D. J.; Baldwin, K. A.; McKay, L. L.

    2000-01-01

    A functional pyc gene was isolated from Lactococcus lactis subsp. lactis C2 and was found to complement a Pyc defect in L. lactis KB4. The deduced lactococcal Pyc protein was highly homologous to Pyc sequences of other bacteria. The pyc gene was also detected in Lactococcus lactis subsp. cremoris and L. lactis subsp. lactis bv. diacetylactis strains.

  3. Environmental Mycobacterium avium subsp. paratuberculosis hosted by free-living amoebae

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis is responsible for paratuberculosis in animals. This disease, leading to an inflammation of the gastrointestinal tract, has a high impact on animal health and an important economic burden. The environmental life cycle of Mycobacterium avium subsp. paratube...

  4. MAO-A inhibition profiles of some benzophenone glucosides from Gentiana verna subsp. pontica

    DEFF Research Database (Denmark)

    Kaya, Duygu; Jäger, Anna; Yalçin, Funda N

    2014-01-01

    Gentiana verna L. subsp. pontica (Soltok.) Hayek, G. pyrenaica L., and G. verna L. subsp. balcanica Pritchard from Turkey were tested for their MAO-A inhibitory effects. A photometric peroxidase linked MAO-A bioassay performed on the H20 extracts prepared from the methanolic extracts of the title...

  5. Preliminary investigation of a mice model of Klebsiella pneumoniae subsp. ozaenae induced pneumonia.

    Science.gov (United States)

    Renois, Fanny; Jacques, Jérôme; Guillard, Thomas; Moret, Hélène; Pluot, Michel; Andreoletti, Laurent; de Champs, Christophe

    2011-11-01

    In the present study, we comparatively assessed the pathophysiological mechanisms developed during lung infection of BALB/C female mice infected by an original wild type Klebsiella pneumoniae subsp. ozaenae strain (CH137) or by a referent subspecies K. pneumoniae. subsp. pneumoniae strain (ATCC10031). The mice infected with 2.10⁶ CFU K. p. subsp. pneumoniae (n = 10) showed transient signs of infection and all of them recovered. All of those infected with 1.10⁶ CFU K. p. subsp. ozaenae (n = 10) developed pneumonia within 24 h and died between 48 and 72 h. Few macrophages, numerous polymorphonuclear cells and lymphocytes were observed in their lungs in opposite to K. p. subsp. pneumoniae. In bronchoalveolar lavage, a significant increase in MIP-2, IL-6, KC and MCP-1 levels was only observed in K. p. subsp. ozaenae infected mice whereas high levels of TNF-α were evidenced with the two subspecies. Our findings indicated a lethal effect of a wild type K. p. subsp. ozaenae strain by acute pneumonia reflecting an insufficient alveolar macrophage response. This model might be of a major interest to comparatively explore the pathogenicity of K. p. subsp ozaenae strains and to further explore the physiopathological mechanisms of gram-negative bacteria induced human pneumonia. Copyright © 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  6. Draft genome sequence of the first human isolate of the ruminant pathogen Mycoplasma capricolum subsp. capricolum

    DEFF Research Database (Denmark)

    Seersholm, Frederik Valeur; Fischer, Anne; Heller, Martin

    2015-01-01

    Mycoplasma capricolum subsp. capricolum is a well-known pathogen of small ruminants. A recent human case of septicemia involving this agent raised the question of its potential pathogenicity to humans. We present the first draft genome sequence of a human Mycoplasma capricolum subsp. capricolum...

  7. Draft Genome Sequence of a Virulent Pectobacterium carotovorum subsp. brasiliense Isolate Causing Soft Rot of Cucumber.

    Science.gov (United States)

    Onkendi, Edward M; Ramesh, Aadi Moolam; Kwenda, Stanford; Naidoo, Sanushka; Moleleki, Lucy

    2016-01-07

    Pectobacterium carotovorum subsp. brasiliense causes soft rot and blackleg diseases on potatoes, ornamentals, and other crops of economic importance. Here, we report a draft genome sequence of a highly virulent P. carotovorum subsp. brasiliense strain, PcbHPI01, isolated from a cucumber in South Africa. Copyright © 2016 Onkendi et al.

  8. Biosystematic studies on Enicostema axillare (Lam.) A. Raynal subsp. Axillare (Gentianaceae) in peninsular India.

    Science.gov (United States)

    Shahina, P M; Nampy, Santhosh

    2014-05-01

    The pantropical genus Enicostema (Gentianaceae) has three species and two sub species world over, namely, E. verticillatum (L.) Engl. (America), E. elizabethae Veldkamp (Madagascar) and E. axillare having 3 subsp. viz., subsp. axillare (Lam.) A. Raynal (India), subsp. latilobum (N.E. Br.) A. Raynal (East Africa) and subsp. littorale (Blume) A. Raynal (Indonesia). The present study aims to delimit the Indian taxa based on field and herbarium studies. Comparative morphology is studied using live as well as consulting wide range of specimens housed at various herbaria. The anatomy of leaf, stem, and root is studied using free hand sections and from epidermal peelings. The seed and pollen morphology are studied under SEM. Information on anatomy, palynology and seed micromorphology of E. axillare subsp. axillare is provided for the first time.

  9. Hospedeiros alternativos de Acidovorax avenae subsp. citrulli Alternative hosts of Acidovorax avenae subsp. citrulli

    Directory of Open Access Journals (Sweden)

    Ana Rosa P. Nascimento

    2004-09-01

    Full Text Available Uma das principais doenças que afeta o meloeiro é a mancha-aquosa, causada pela bactéria Acidovorax avenae subsp. citrulli (Aac. Visando conhecer hospedeiros alternativos de Aac, plantas no estágio de primeiras folhas definitivas, de várias espécies/cultivares, incluindo cucurbitáceas, solanáceas, gramíneas, leguminosas e caricáceas, foram inoculadas pela atomização da parte aérea com suspensão dos isolados Aac 1.49 e Aac 12.13, oriundos de melão e melancia, respectivamente. A suscetibilidade das plantas aos isolados foi avaliada pelo período de incubação (PI e incidência da doença (INC. Caupi, feijão, fumo e milho não apresentaram sintomas. Os menores PIs foram observados em cucurbitáceas (3,0 d, com exceção da bucha (6,83 d. Incidências da doença acima de 90% foram observadas em cucurbitáceas, excetuando a bucha e em solanáceas, para ambos os isolados de Aac. Em outro experimento, frutos de abóbora, abobrinha, berinjela, mamão, maxixe, melancia, melão, pepino, pimentão e tomate foram analisados quanto à suscetibilidade aos isolados Aac 1.49 e Aac 12.13. Os frutos foram inoculados pelo método de injeção subepidérmica, determinando-se PI, INC e severidade, avaliada pelo diâmetro da lesão externa (DLE e profundidade da lesão (PL. Menores PIs (2,0 d foram detectados em frutos de mamão, melancia, melão e pimentão. Incidência de 100% foi observada em todos os frutos inoculados, com exceção da abobrinha (93,75% e da abóbora (34,37%. Maiores DLEs foram observados em pepino (1,47 cm para o isolado Aac 1.49 e em melancia (1,60 cm e melão (1,07 cm para Aac 12.13. As maiores PL foram constatadas em melancia (1,72 e 0,75 cm respectivamente para Aac 1.49 e Aac 12.13. Frutos de berinjela não apresentaram sintomas externos embora as lesões internas tenham sido profundas.One of the most important melon diseases is the bacterial blotch caused by Acidovorax avenae subsp. citrulli (Aac. Alternative hosts of this

  10. Draft Genome Sequences of Three Pectobacterium Strains Causing Blackleg of Potato: P. carotovorum subsp. brasiliensis ICMP 19477, P. atrosepticum ICMP 1526, and P. carotovorum subsp. carotovorum UGC32.

    Science.gov (United States)

    Panda, Preetinanda; Fiers, Mark W E J; Lu, Ashley; Armstrong, Karen F; Pitman, Andrew R

    2015-08-06

    Blackleg is a disease caused by several species of Pectobacterium that results in losses to potato crops worldwide. Here, we report the draft genomes of three taxonomically and geographically distinct blackleg-causing strains of Pectobacterium: P. carotovorum subsp. brasiliensis ICMP 19477, P. atrosepticum ICMP 1526, and P. carotovorum subsp. carotovorum UGC32. Comparison of these genomes will support the identification of common traits associated with their capacity to cause blackleg. Copyright © 2015 Panda et al.

  11. Investigation of Phenolic Compounds and Antioxidant Activity of Mentha spicata L. subsp. spicata and M. longifolia (L.) L. subsp. typhoides (Briq.) Harley Decoction and Infusion

    OpenAIRE

    ÖZER, Züleyha

    2018-01-01

    In present study, we report phenolic compounds and antioxidant activity of decoctionand infusion of Mentha spicata L. subsp. spicataand M. longifolia (L.) L. subsp. typhoides (Briq.) Harley. The quantitativeamounts of the phenolic contents were determined by LC-MS/MS.  The main compounds and amounts of M. spicata weredetermined as follow for decoction; caffeic acid, quercetagetin-3,6-dimethyletherand penduletin (4126.6; 2141.5; 1472.7 mg/kg dried herba, respectively), for infusion;fumaric aci...

  12. Relationship of Bacillus amyloliquefaciens clades associated with strains DSM 7T and FZB42T: a proposal for Bacillus amyloliquefaciens subsp. amyloliquefaciens subsp. nov. and Bacillus amyloliquefaciens subsp. plantarum subsp. nov. based on complete genome sequence comparisons.

    Science.gov (United States)

    Borriss, Rainer; Chen, Xiao-Hua; Rueckert, Christian; Blom, Jochen; Becker, Anke; Baumgarth, Birgit; Fan, Ben; Pukall, Rüdiger; Schumann, Peter; Spröer, Cathrin; Junge, Helmut; Vater, Joachim; Pühler, Alfred; Klenk, Hans-Peter

    2011-08-01

    The whole-genome-sequenced rhizobacterium Bacillus amyloliquefaciens FZB42(T) (Chen et al., 2007) and other plant-associated strains of the genus Bacillus described as belonging to the species Bacillus amyloliquefaciens or Bacillus subtilis are used commercially to promote the growth and improve the health of crop plants. Previous investigations revealed that a group of strains represented a distinct ecotype related to B. amyloliquefaciens; however, the exact taxonomic position of this group remains elusive (Reva et al., 2004). In the present study, we demonstrated the ability of a group of Bacillus strains closely related to strain FZB42(T) to colonize Arabidopsis roots. On the basis of their phenotypic traits, the strains were similar to Bacillus amyloliquefaciens DSM 7(T) but differed considerably from this type strain in the DNA sequences of genes encoding 16S rRNA, gyrase subunit A (gyrA) and histidine kinase (cheA). Phylogenetic analysis performed with partial 16S rRNA, gyrA and cheA gene sequences revealed that the plant-associated strains of the genus Bacillus, including strain FZB42(T), formed a lineage, which could be distinguished from the cluster of strains closely related to B. amyloliquefaciens DSM 7(T). DNA-DNA hybridizations (DDH) performed with genomic DNA from strains DSM 7(T) and FZB42(T) yielded relatedness values of 63.7-71.2 %. Several methods of genomic analysis, such as direct whole-genome comparison, digital DDH and microarray-based comparative genomichybridization (M-CGH) were used as complementary tests. The group of plant-associated strains could be distinguished from strain DSM 7(T) and the type strain of B. subtilis by differences in the potential to synthesize non-ribosomal lipopeptides and polyketides. Based on the differences found in the marker gene sequences and the whole genomes of these strains, we propose two novel subspecies, designated B. amyloliquefaciens subsp. plantarum subsp. nov., with the type strain FZB42(T) ( = DSM

  13. Capsular exopolysaccharide biosynthesis gene of Propionibacterium freudenreichii subsp. shermanii.

    Science.gov (United States)

    Deutsch, Stéphanie-Marie; Falentin, Hélène; Dols-Lafargue, Marguerite; Lapointe, Gisèle; Roy, Denis

    2008-07-31

    In the dairy industry, exopolysaccharides (EPS) contribute to improving the texture and viscosity of cheese and yoghurt and also receive increasing attention because of their beneficial properties for health. For lactic acid bacteria, the production of EPS is well studied. However, for dairy propionibacteria the biosynthesis of EPS is poorly documented. A polysaccharide synthase-encoding gene was identified in the genome of Propionibacterium freudenreichii subsp. shermanii TL 34 (CIP 103027). This gene best aligns with Tts, the polysaccharide synthase gene of Streptococcus pneumoniae type 37 that is responsible for the production of a beta-glucan capsular polysaccharide. PCR amplification showed the presence of an internal fragment of this gene in twelve strains of P. freudenreichii subsp. shermanii with a ropy phenotype in YEL+ medium. The gene sequence is highly conserved, as less than 1% of nucleotides differed among the 10 strains containing the complete gtf gene. The same primers failed to detect the gene in Propionibacterium acidipropionici strain TL 47, which is known to excrete exopolysaccharides in milk. The presence of (1-->3, 1-->2)-beta-d-glucan capsule was demonstrated for 7 out of 12 strains by agglutination with a S. pneumoniae-type 37-specific antiserum. The presence of mRNA corresponding to the gene was detected by RT-PCR in three strains at both exponential and stationary growth phases. This work represents the first identification of a polysaccharide synthase gene of P. freudenreichii, and further studies will be undertaken to elucidate the role of capsular EPS.

  14. Role of Blossoms in Watermelon Seed Infestation by Acidovorax avenae subsp. citrulli.

    Science.gov (United States)

    Walcott, R R; Gitaitis, R D; Castro, A C

    2003-05-01

    ABSTRACT The role of watermelon blossom inoculation in seed infestation by Acidovorax avenae subsp. citrulli was investigated. Approximately 98% (84/87) of fruit developed from blossoms inoculated with 1 x 10(7) or 1 x 10(9) CFU of A. avenae subsp. citrulli per blossom were asymptomatic. Using immunomagnetic separation and the polymerase chain reaction, A. avenae subsp. citrulli was detected in 44% of the seed lots assayed, despite the lack of fruit symptoms. Furthermore, viable colonies were recovered from 31% of the seed lots. Of these lots, 27% also yielded seedlings expressing bacterial fruit blotch symptoms when planted under conditions of 30 degrees C and 90% relative humidity. A. avenae subsp. citrulli was detected and recovered from the pulp of 33 and 19%, respectively, of symptomless fruit whose blossoms were inoculated with A. avenae subsp. citrulli. The ability to penetrate watermelon flowers was not unique to A. avenae subsp. citrulli, because blossoms inoculated with Pantoea ananatis also resulted in infested seed and pulp. The data indicate that watermelon blossoms are a potential site of ingress for fruit and seed infestation by A. avenae subsp. citrulli.

  15. Environmental Mycobacterium avium subsp. paratuberculosis Hosted by Free-Living Amoebae

    Directory of Open Access Journals (Sweden)

    Ascel Samba-Louaka

    2018-02-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis is responsible for paratuberculosis in animals. This disease, leading to an inflammation of the gastrointestinal tract, has a high impact on animal health and an important economic burden. The environmental life cycle of M. avium subsp. paratuberculosis is poorly understood and several studies suggest that free-living amoebae (FLA might be a potential environmental host. FLA are protozoa found in water and soil that are described as reservoirs of pathogenic and non-pathogenic bacteria in the environment. Indeed, bacteria able to survive within these amoebae would survive phagocytosis from immune cells. In this study, we assessed the in vitro interactions between several strains of M. avium subsp. paratuberculosis and Acanthamoeba castellanii. The results indicate that the bacteria were able to grow within the amoeba and that they can survive for several days within their host. To explore the presence of M. avium subsp. paratuberculosis in environmental amoebae, we sampled water from farms positive for paratuberculosis. A M. avium subsp. paratuberculosis strain was detected within an environmental amoeba identified as related to the poorly described Rosculus genus. The bacterial strain was genotyped, showing that it was similar to previous infectious strains isolated from cattle. In conclusion, we described that various M. avium subsp. paratuberculosis strains were able to grow within amoebae and that these bacteria could be found on farm within amoebae isolated from the cattle environment. It validates that infected amoebae might be a reservoir and vector for the transmission of M. avium subsp. paratuberculosis.

  16. Lactobacillus paracasei subsp. paracasei B21060 suppresses human T-cell proliferation.

    Science.gov (United States)

    Peluso, Ilaria; Fina, Daniele; Caruso, Roberta; Stolfi, Carmine; Caprioli, Flavio; Fantini, Massimo Claudio; Caspani, Giorgio; Grossi, Enzo; Di Iorio, Laura; Paone, Francesco Maria; Pallone, Francesco; Monteleone, Giovanni

    2007-04-01

    Recent studies have shown that probiotics are beneficial in T-cell-mediated inflammatory diseases. The molecular mechanism by which probiotics work remains elusive, but accumulating evidence indicates that probiotics can modulate immune cell responses. Since T cells express receptors for bacterial products or components, we examined whether different strains of lactobacilli directly regulate the functions of human T cells. CD4(+) T cells were isolated from blood and intestinal lamina propria (LP) of normal individuals and patients with inflammatory bowel disease (IBD). Mononuclear cells were also isolated from Peyer's patches. Cells were activated with anti-CD3/CD2/CD28 in the presence or absence of Lactobacillus paracasei subsp. paracasei B21060, L. paracasei subsp. paracasei F19, or L. casei subsp. casei DG. Cell proliferation and death, Foxp3, intracellular pH, and cytokine production were evaluated by flow cytometry. We showed that L. paracasei subsp. paracasei B21060 but neither L. paracasei subsp. paracasei F19 nor L. casei subsp. casei DG inhibited blood CD4(+) T-cell growth. This effect was associated with no change in cell survival, expression of Foxp3, or production of gamma interferon, interleukin-4 (IL-4), IL-5, and IL-10. L. paracasei subsp. paracasei B21060-mediated blockade of CD4(+) T-cell proliferation required a viable bacterium and was associated with decreased MCT-1 expression and low intracellular pH. L. paracasei subsp. paracasei B21060 also inhibited the growth of Peyer's patch mononuclear cells, normal lymphocytes, and IBD CD4(+) LP lymphocytes without affecting cytokine production. The data show that L. paracasei subsp. paracasei B21060 blocks T-cell growth, thus suggesting a mechanism by which these probiotics could interfere with T-cell-driven immune responses.

  17. Proposal to reclassify Brenneria quercina (Hildebrand and Schroth 1967) Hauben et al. 1999 into a new genus, Lonsdalea gen. nov., as Lonsdalea quercina comb. nov., descriptions of Lonsdalea quercina subsp. quercina comb. nov., Lonsdalea quercina subsp. iberica subsp. nov. and Lonsdalea quercina subsp. britannica subsp. nov., emendation of the description of the genus Brenneria, reclassification of Dickeya dieffenbachiae as Dickeya dadantii subsp. dieffenbachiae comb. nov., and emendation of the description of Dickeya dadantii.

    Science.gov (United States)

    Brady, Carrie L; Cleenwerck, Ilse; Denman, Sandra; Venter, Stephanus N; Rodríguez-Palenzuela, Pablo; Coutinho, Teresa A; De Vos, Paul

    2012-07-01

    Bacterial isolates from oak trees in Spain and Britain, showing symptoms of bark canker and Acute Oak Decline (AOD), respectively, were examined by a polyphasic approach. Both 16S rRNA gene sequencing and multilocus sequence analysis (MLSA), based on partial sequences of gyrB, rpoB, infB and atpD genes, revealed that the isolates were separated into two genetic groups according to their origin. Their closest phylogenetic relative was Brenneria quercina, the causal agent of drippy nut disease of oak, which clustered distant to the other species of the genus Brenneria. MLSA data for species of the genera Brenneria, Pectobacterium, Dickeya, Erwinia, Pantoea and Samsonia confirmed the polyphyletic nature of the genus Brenneria and indicated synonymy of Dickeya dadantii and Dickeya dieffenbachiae. DNA-DNA hybridization experiments confirmed this synonymy and also revealed DNA-DNA relatedness values of 58-73% between the new oak isolates and B. quercina. Phenotypic and/or chemotaxonomic methods allowed B. quercina and the two genetic groups of new oak isolates to be discriminated from other recognized species of the genus Brenneria and from members of the closely related genera Dickeya, Pectobacterium and Samsonia. Based on the data obtained, the following taxonomic proposals are made: (1) reclassification of B. quercina as the type species of a novel genus, Lonsdalea gen. nov., as Lonsdalea quercina comb. nov. (type strain LMG 2724(T)=ATCC 29281(T)=CCUG 48867(T)=CFBP 3617(T)=CIP 105201(T)=DSM 4561(T)=ICMP 1845(T)), (2) classification of the oak isolates as Lonsdalea quercina subsp. iberica subsp. nov. (type strain LMG26264(T)=NCPPB 4490(T)) and Lonsdalea quercina subsp. britannica subsp. nov. (type strain LMG 26267(T)=NCPPB 4481(T)) and leading to the automatic creation of Lonsdalea quercina subsp. quercina subsp. nov. (type strain LMG 2724(T)=ATCC 29281(T)), (3) emendation of the description of the genus Brenneria, and (4) reclassification of Dickeya dieffenbachiae as

  18. Stability evaluation of freeze-dried Lactobacillus paracasei subsp. tolerance and Lactobacillus delbrueckii subsp. bulgaricus in oral capsules.

    Science.gov (United States)

    Jalali, M; Abedi, D; Varshosaz, J; Najjarzadeh, M; Mirlohi, M; Tavakoli, N

    2012-01-01

    Freeze-drying is a common preservation technology in the pharmaceutical industry. Various studies have investigated the effect of different cryoprotectants on probiotics during freeze-drying. However, information on the effect of cryoprotectants on the stability of some Lactobacillus strains during freeze-drying seems scarce. Therefore, the aim of the present study was to establish production methods for preparation of oral capsule probiotics containing Lactobacillus paracasei subsp. tolerance and Lactobacillus delbrueckii subsp. Bulgaricus. It was also of interest to examine the effect of various formulations of cryoprotectant media containing skim milk, trehalose and sodium ascorbate on the survival rate of probiotic bacteria during freeze-drying at various storage temperatures. Without any cryoprotectant, few numbers of microorganisms survived. However, microorganisms tested maintained higher viability after freeze-drying in media containing at least one of the cryoprotectants. Use of skim milk in water resulted in an increased viability after lyophilization. Media with a combination of trehalose and skim milk maintained a higher percentage of live microorganisms, up to 82%. In general, bacteria retained a higher number of viable cells in capsules containing freeze-dried bacteria with sodium ascorbate after three months of storage. After this period, a marked decline was observed in all samples stored at 23°C compared to those stored at 4°C. The maximum survival rate (about 72-76%) was observed with media containing 6% skim milk, 8% trehalose and 4% sodium ascorbate.

  19. The Type III Secretion System Effector SeoC of Salmonella enterica subsp. salamae and S. enterica subsp. arizonae ADP-Ribosylates Src and Inhibits Opsonophagocytosis.

    Science.gov (United States)

    Pollard, Dominic J; Young, Joanna C; Covarelli, Valentina; Herrera-León, Silvia; Connor, Thomas R; Fookes, Maria; Walker, Danielle; Echeita, Aurora; Thomson, Nicholas R; Berger, Cedric N; Frankel, Gad

    2016-12-01

    Salmonella species utilize type III secretion systems (T3SSs) to translocate effectors into the cytosol of mammalian host cells, subverting cell signaling and facilitating the onset of gastroenteritis. In this study, we compared a draft genome assembly of Salmonella enterica subsp. salamae strain 3588/07 against the genomes of S. enterica subsp. enterica serovar Typhimurium strain LT2 and Salmonella bongori strain 12419. S. enterica subsp. salamae encodes the Salmonella pathogenicity island 1 (SPI-1), SPI-2, and the locus of enterocyte effacement (LEE) T3SSs. Though several key S Typhimurium effector genes are missing (e.g., avrA, sopB, and sseL), S. enterica subsp. salamae invades HeLa cells and contains homologues of S. bongori sboK and sboC, which we named seoC SboC and SeoC are homologues of EspJ from enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC, respectively), which inhibit Src kinase-dependent phagocytosis by ADP-ribosylation. By screening 73 clinical and environmental Salmonella isolates, we identified EspJ homologues in S. bongori, S. enterica subsp. salamae, and Salmonella enterica subsp. arizonae The β-lactamase TEM-1 reporter system showed that SeoC is translocated by the SPI-1 T3SS. All the Salmonella SeoC/SboC homologues ADP-ribosylate Src E310 in vitro Ectopic expression of SeoC/SboC inhibited phagocytosis of IgG-opsonized beads into Cos-7 cells stably expressing green fluorescent protein (GFP)-FcγRIIa. Concurrently, S. enterica subsp. salamae infection of J774.A1 macrophages inhibited phagocytosis of beads, in a seoC-dependent manner. These results show that S. bongori, S. enterica subsp. salamae, and S. enterica subsp. arizonae share features of the infection strategy of extracellular pathogens EPEC and EHEC and shed light on the complexities of the T3SS effector repertoires of Enterobacteriaceae. Copyright © 2016 Pollard et al.

  20. Molecular Characterization of Three Lactobacillus delbrueckii subsp. bulgaricus Phages

    Science.gov (United States)

    Casey, Eoghan; Mahony, Jennifer; O'Connell-Motherway, Mary; Bottacini, Francesca; Cornelissen, Anneleen; Neve, Horst; Heller, Knut J.; Noben, Jean-Paul; Dal Bello, Fabio

    2014-01-01

    In this study, three phages infecting Lactobacillus delbrueckii subsp. bulgaricus, named Ld3, Ld17, and Ld25A, were isolated from whey samples obtained from various industrial fermentations. These phages were further characterized in a multifaceted approach: (i) biological and physical characterization through host range analysis and electron microscopy; (ii) genetic assessment through genome analysis; (iii) mass spectrometry analysis of the structural components of the phages; and (iv), for one phage, transcriptional analysis by Northern hybridization, reverse transcription-PCR, and primer extension. The three obtained phage genomes display high levels of sequence identity to each other and to genomes of the so-called group b L. delbrueckii phages c5, LL-Ku, and phiLdb, where some of the observed differences are believed to be responsible for host range variations. PMID:25002431

  1. Purulent pericarditis and pneumonia caused by Streptococcus equi subsp. zooepidemicus.

    Science.gov (United States)

    Held, Jürgen; Schmitz, Roland; van der Linden, Mark; Nührenberg, Thomas; Häcker, Georg; Neumann, Franz-Josef

    2014-02-01

    Purulent pericarditis is a life-threatening disease that usually manifests following bacteraemia or through spreading from an intrathoracic focus. Only a few cases of this disease have been reported with Lancefield group C streptococci as aetiological agents, and the primary focus in these infections remains unknown. We report a case of purulent pericarditis with septic and cardiogenic shock, caused by Streptococcus equi subsp. zooepidemicus (group C) in a 51-year-old patient. The pathogen was possibly contracted through contact with horses. Most probably, it caused initially pneumonia before spreading to the pericardium, either directly or via the bloodstream. A combined therapeutic approach, consisting of antibiotic therapy and repeated pericardial drainage, was necessary to ensure a clinical cure. After discharge, long-term follow-up for development of constrictive pericarditis is considered mandatory.

  2. Survival of pathogenic Mycobacterium abscessus subsp. massiliense in Acanthamoeba castellanii.

    Science.gov (United States)

    da Silva, Joas L; Nguyen, Jan; Fennelly, Kevin P; Zelazny, Adrian M; Olivier, Kenneth N

    2018-01-01

    We used an amoeba model to study the intracellular growth and cytotoxicity of clinical strains of Mycobacterium abscessus subsp. massiliense (Mabsm) isolated from 2 patients (one with cystic fibrosis, the other one with idiopathic bronchiectasis) during the early (smooth colonies) and late stage (rough colonies) of chronic pulmonary infection. Acanthamoeba castellanii were infected with Mabsm (MOI 100) and samples collected every 24 h for 72 h. Results showed Mabsm is able to survive in trophozoites and persist in cysts for at least 7 days. Late Mabsm demonstrated higher cytotoxicity toward A. castellanii when compared to early strains. A. castellanii is a useful in vitro host model to study infection of Mabsm clinical isolates. Published by Elsevier Masson SAS.

  3. A Mycobacterium avium subsp. paratuberculosis predicted serine protease is associated with acid stress and intraphagosomal survival

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular pathogen that persists inside host macrophages despite severe oxidative stress and nutrient deprivation. Intrabacterial pH homeostasis is vital to pathogenic mycobacteria to preserve cellular biological processes and stability of ...

  4. Composition and potency characterization of Mycobacterium avium subsp. paratuberculosis purified protein derivatives

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) purified protein derivatives (PPDs) are immunologic reagents prepared from cultured filtrates of the type strain ATCC 19698. Traditional production consists of floating culture incubation at 37oC, organism inactivation by autoclaving, coarse filtrat...

  5. Influence of ions on growth and production of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772

    NARCIS (Netherlands)

    Grobben, G.J.; Boels, I.C.; Sikkema, J.; Smith, M.R.; Bont, de J.A.M.

    2000-01-01

    Several lactic acid bacteria produce exopolysaccharides (EPS), either attached to the cell wall or excreted into the environment as slime material. EPS produced by Lactobacillus delbrueckii subsp. bulgaricus (Lb. bulgaricus) and Streptococcus thermophilus play an important role in improving the

  6. Klebsiella alba is a later heterotypic synonym of Klebsiella quasipneumoniae subsp. similipneumoniae.

    Science.gov (United States)

    Li, Chun Yan; Zhou, Yuan Liang; Ji, Jing; Gu, Chun Tao

    2016-06-01

    The taxonomic position of Klebsiella alba was re-examined. The reconstructed phylogenetic tree based on multilocus sequence analysis showed that Klebsiella alba LMG 24441T (=KCTC 12878T) was closely related to Klebsiella quasipneumoniae subsp. similipneumoniae 07A044T, showing 99.5-100 % similarities for fusA, gapA, gyrA, leuS, pyrG, rpoB, rpoB2, atpD, gyrB and infB gene sequences and concatenated partial fusA, gapA, gyrA, leuS, pyrG, rpoB2, atpD, gyrB and infB gene sequences. High sequence similarities between Klebsiella alba LMG 24441T and Klebsiella quasipneumoniae subsp. similipneumoniae 07A044T indicated that they have the same taxonomic position. Klebsiellaalba was reclassified as Klebsiellaquasipneumoniae subsp. similipneumoniae and Klebsiella alba is a later heterotypic synonym of Klebsiella quasipneumoniae subsp. similipneumoniae.

  7. Mean effective sensitivity for Mycobacterium avium subsp paratuberculosis infection in cattle herds

    DEFF Research Database (Denmark)

    Kirkeby, Carsten; Græsbøll, Kaare; Hisham Beshara Halasa, Tariq

    2015-01-01

    Background: Mycobacterium avium subsp. paratuberculosis (MAP) infections in cattle are generally challenging to detect and cost-effective test strategies are consequently difficult to identify. MAP-specific antibody ELISAs for milk and serum are relatively inexpensive, but their utility is influe......Background: Mycobacterium avium subsp. paratuberculosis (MAP) infections in cattle are generally challenging to detect and cost-effective test strategies are consequently difficult to identify. MAP-specific antibody ELISAs for milk and serum are relatively inexpensive, but their utility...

  8. [Identification and phylogenetic analysis of one strain of Lactobacillus delbrueckii subsp. bulgaricus separated from yoghourt].

    Science.gov (United States)

    Wang, Chuan; Zhang, Chaowu; Pei, Xiaofang; Liu, Hengchuan

    2007-11-01

    For being further applied and studied, one strain of Lactobacillus delbrueckii subsp. bulgaricus (wch9901) separated from yoghourt which had been identified by phenotype characteristic analysis was identified by 16S rDNA and phylogenetic analyzed. The 16S rDNA of wch9901 was amplified with the genomic DNA of wch9901 as template, and the conservative sequences of the 16S rDNA as primers. Inserted 16S rDNA amplified into clonal vector pGEM-T under the function of T4 DNA ligase to construct recombined plasmid pGEM-wch9901 16S rDNA. The recombined plasmid was identified by restriction enzyme digestion, and the eligible plasmid was presented to sequencing company for DNA sequencing. Nucleic acid sequence was blast in GenBank and phylogenetic tree was constructed using neighbor-joining method of distance methods by Mega3.1 soft. Results of blastn showed that the homology of 16S rDNA of wch9901 with the 16S rDNA of Lactobacillus delbrueckii subsp. bulgaricus strains was higher than 96%. On the phylogenetic tree, wch9901 formed a separate branch and located between Lactobacillus delbrueckii subsp. bulgaricus LGM2 evolution branch and another evolution branch which was composed of Lactobacillus delbrueckii subsp. bulgaricus DL2 evolution cluster and Lactobacillus delbrueckii subsp. bulgaricus JSQ evolution cluster. The distance between wch9901 evolution branch and Lactobacillus delbrueckii subsp. bulgaricus LGM2 evolution branch was the closest. wch9901 belonged to Lactobacillus delbrueckii subsp. bulgaricus. wch9901 showed the closest evolution relationship to Lactobacillus delbrueckii subsp. bulgaricus LGM2.

  9. Epidemiology of Danish Aeromonas salmonicida subsp salmonicida in Fish Farms Using Whole Genome Sequencing

    DEFF Research Database (Denmark)

    Bartkova, Simona; Leekitcharoenphon, Pimlapas; Aarestrup, Frank Møller

    2017-01-01

    for presence of plasmids, virulence and iron acquisition proteins, genomic islands, and antibiotic resistance genes. Single Nucleotide Polymorphisms were aligned and subjected to Bayesian temporal phylogenetic and maximum likelihood tree reconstruction using the published genome of A. salmonicida subsp....... salmonicida A449 as reference. Bayesian temporal phylogenetic reconstruction suggests that four major introductions of A. salmonicida subsp. salmonicida into Denmark have occurred. The introductions correlate with the freshwater and subsequent seawater expansion of rainbow trout production. Initial...

  10. Inside the adaptation process of Lactobacillus delbrueckii subsp. lactis to bile

    OpenAIRE

    Burns, Patricia; Sánchez García, Borja; Vinderola, Gabriel; Ruas-Madiedo, Patricia; Ruíz García, Lorena; Margolles Barros, Abelardo; Reinheimer, Jorge A.; González de los Reyes-Gavilán, Clara

    2010-01-01

    Progressive adaptation to bile might render some lactobacilli able to withstand physiological bile salt concentrations. In this work, the adaptation to bile was evaluated on previously isolated dairy strains of Lactobacillus delbrueckii subsp. lactis 200 and L. delbrueckii subsp. lactis 200+, a strain derived thereof with stable bile-resistant phenotype. The adaptation to bile was obtained by comparing cytosolic proteomes of both strains grown in the presence or absence of bile. Proteomics we...

  11. Genomic variations of Mycoplasma capricolum subsp capripneumoniae detected by amplified fragment length polymorphism (AFLP) analysis

    DEFF Research Database (Denmark)

    Kokotovic, Branko; Bolske, G.; Ahrens, Peter

    2000-01-01

    The genetic diversity of Mycoplasma capricolum subsp. capripneumoniae strains based on determination of amplified fragment length polymorphisms (AFLP) is described. AFLP fingerprints of 38 strains derived from different countries in Africa and the Middle East consisted of over 100 bands in the size...... found by 16S rDNA analysis. The present data support previous observations regarding genetic homogeneity of M. capricolum subsp. capripneumoniae, and confirm the two evolutionary lines of descent found by analysis of 16S rRNA genes....

  12. Complete Genome Sequence of Phytopathogenic Pectobacterium carotovorum subsp. carotovorum Bacteriophage PP1

    OpenAIRE

    Lee, Ju-Hoon; Shin, Hakdong; Ji, Samnyu; Malhotra, Shweta; Kumar, Mukesh; Ryu, Sangryeol; Heu, Sunggi

    2012-01-01

    Pectobacterium carotovorum subsp. carotovorum is a phytopathogen causing soft rot disease on diverse plant species. To control this plant pathogen, P. carotovorum subsp. carotovorum-targeting bacteriophage PP1 was isolated and its genome was completely sequenced to develop a novel biocontrol agent. Interestingly, the 44,400-bp genome sequence does not encode any gene involved in the formation of lysogen, suggesting that this phage may be very useful as a biocontrol agent because it does not m...

  13. Decreased toxicity of Bacillus thuringiensis subsp. israelensis to mosquito larvae after contact with leaf litter.

    Science.gov (United States)

    Tetreau, Guillaume; Stalinski, Renaud; Kersusan, Dylann; Veyrenc, Sylvie; David, Jean-Philippe; Reynaud, Stéphane; Després, Laurence

    2012-08-01

    Bacillus thuringiensis subsp. israelensis is a bacterium producing crystals containing Cry and Cyt proteins, which are toxic for mosquito larvae. Nothing is known about the interaction between crystal toxins and decaying leaf litter, which is a major component of several mosquito breeding sites and represents an important food source. In the present work, we investigated the behavior of B. thuringiensis subsp. israelensis toxic crystals sprayed on leaf litter. In the presence of leaf litter, a 60% decrease in the amount of Cyt toxin detectable by immunology (enzyme-linked immunosorbent assays [ELISAs]) was observed, while the respective proportions of Cry toxins were not affected. The toxicity of Cry toxins toward Aedes aegypti larvae was not affected by leaf litter, while the synergistic effect of Cyt toxins on all B. thuringiensis subsp. israelensis Cry toxins was decreased by about 20% when mixed with leaf litter. The toxicity of two commercial B. thuringiensis subsp. israelensis strains (VectoBac WG and VectoBac 12AS) and a laboratory-produced B. thuringiensis subsp. israelensis strain decreased by about 70% when mixed with leaf litter. Taken together, these results suggest that Cyt toxins interact with leaf litter, resulting in a decreased toxicity of B. thuringiensis subsp. israelensis in litter-rich environments and thereby dramatically reducing the efficiency of mosquitocidal treatments.

  14. Decreased Toxicity of Bacillus thuringiensis subsp. israelensis to Mosquito Larvae after Contact with Leaf Litter

    Science.gov (United States)

    Stalinski, Renaud; Kersusan, Dylann; Veyrenc, Sylvie; David, Jean-Philippe; Reynaud, Stéphane; Després, Laurence

    2012-01-01

    Bacillus thuringiensis subsp. israelensis is a bacterium producing crystals containing Cry and Cyt proteins, which are toxic for mosquito larvae. Nothing is known about the interaction between crystal toxins and decaying leaf litter, which is a major component of several mosquito breeding sites and represents an important food source. In the present work, we investigated the behavior of B. thuringiensis subsp. israelensis toxic crystals sprayed on leaf litter. In the presence of leaf litter, a 60% decrease in the amount of Cyt toxin detectable by immunology (enzyme-linked immunosorbent assays [ELISAs]) was observed, while the respective proportions of Cry toxins were not affected. The toxicity of Cry toxins toward Aedes aegypti larvae was not affected by leaf litter, while the synergistic effect of Cyt toxins on all B. thuringiensis subsp. israelensis Cry toxins was decreased by about 20% when mixed with leaf litter. The toxicity of two commercial B. thuringiensis subsp. israelensis strains (VectoBac WG and VectoBac 12AS) and a laboratory-produced B. thuringiensis subsp. israelensis strain decreased by about 70% when mixed with leaf litter. Taken together, these results suggest that Cyt toxins interact with leaf litter, resulting in a decreased toxicity of B. thuringiensis subsp. israelensis in litter-rich environments and thereby dramatically reducing the efficiency of mosquitocidal treatments. PMID:22610426

  15. Effects of six substances on the growth and freeze-drying of Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Chen, He; Huang, Jie; Shi, Xiaoyu; Li, Yichao; Liu, Yu

    2017-01-01

    The efficacy of Lactobacillus delbrueckii subsp. bulgaricus as starter cultures for the dairy industry depends largely on the number of viable and active cells. Freeze-drying is the most convenient and successful method to preserve the bacterial cells. However, not all strains survived during freeze-drying. The effects of six substances including NaCl, sorbitol, mannitol, mannose, sodium glutamate, betaine added to the MRS medium on the growth and freeze-drying survival rate and viable counts of Lb. delbrueckii subsp. bulgaricus were studied through a single-factor test and Plackett-Burman design. Subsequently, the optimum freeze-drying conditions of Lb. delbrueckii subsp. bulgaricus were determined. Lb. delbrueckii subsp. bulgaricus survival rates were up to the maximum of 42.7%, 45.4%, 23.6%, while the concentrations of NaCl, sorbitol, sodium glutamate were 0.6%, 0.15%, 0.09%, respectively. In the optimum concentration, the viable counts in broth is 6.1, 6.9, 5.13 (×108 CFU/mL), respectively; the viable counts in freeze-drying power are 3.09, 5.2, 2.7 (×1010 CFU/g), respectively. Three antifreeze factors including NaCl, sorbitol, sodium glutamate have a positive effect on the growth and freeze-drying of Lb. delbrueckii subsp. bulgaricus. The results are beneficial for developing Lb. delbrueckii subsp. bulgaricus.

  16. Characterization of pneumonia due to Streptococcus equi subsp. zooepidemicus in dogs.

    Science.gov (United States)

    Priestnall, Simon L; Erles, Kerstin; Brooks, Harriet W; Cardwell, Jacqueline M; Waller, Andrew S; Paillot, Romain; Robinson, Carl; Darby, Alistair C; Holden, Matthew T G; Schöniger, Sandra

    2010-11-01

    Streptococcus equi subsp. zooepidemicus has been linked to cases of acute fatal pneumonia in dogs in several countries. Outbreaks can occur in kenneled dog populations and result in significant levels of morbidity and mortality. This highly contagious disease is characterized by the sudden onset of clinical signs, including pyrexia, dyspnea, and hemorrhagic nasal discharge. The pathogenesis of S. equi subsp. zooepidemicus infection in dogs is poorly understood. This study systematically characterized the histopathological changes in the lungs of 39 dogs from a large rehoming shelter in London, United Kingdom; the dogs were infected with S. equi subsp. zooepidemicus. An objective scoring system demonstrated that S. equi subsp. zooepidemicus caused pneumonia in 26/39 (66.7%) dogs, and most of these dogs (17/26 [65.4%]) were classified as severe fibrino-suppurative, necrotizing, and hemorrhagic. Three recently described superantigen genes (szeF, szeN, and szeP) were detected by PCR in 17/47 (36.2%) of the S. equi subsp. zooepidemicus isolates; however, there was no association between the presence of these genes and the histopathological score. The lungs of S. equi subsp. zooepidemicus-infected dogs with severe respiratory signs and lung pathology did however have significantly higher mRNA levels of the proinflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and interleukin 8 (IL-8) than in uninfected controls, suggesting a role for an exuberant host immune response in the pathogenesis of this disease.

  17. Pork meat as a potential source of Salmonella enterica subsp. arizonae infection in humans.

    Science.gov (United States)

    Evangelopoulou, Grammato; Kritas, Spyridon; Govaris, Alexander; Burriel, Angeliki R

    2014-03-01

    Salmonella enterica subsp. arizonae was isolated from 13 of 123 slaughtered pigs in central Greece. The samples cultured were feces, ileum tissue, mesenteric lymph nodes, and gallbladder swabs. A total of 74 isolates from 492 samples were identified as Salmonella spp. by use of standard laboratory culture media and two commercial micromethods and by use of a polyvalent slide agglutination test for the detection of O and H antigens. Among them were 19 (25.68%) suspected to be S. enterica subsp. arizonae according to analysis with standard laboratory culture media. Of those, 14 were identified as S. enterica subsp. arizonae by the API 20E (bioMérieux, France) and the Microgen GnA+B-ID (Microgen Bioproducts, Ltd., United Kingdom) identification systems. All the isolates were tested for resistance to 23 antimicrobials. Strains identified as S. enterica subsp. arizonae were resistant to 17 (70.8%) antibiotics. The highest proportions of resistance were observed for sulfamethoxazole-trimethoprim (71.4%), tetracycline (71.4%), ampicillin (64.3%), and amoxicillin (57.1%). Two isolates were resistant to aztreonam (7.1%) and tigecycline (7.1%), used only for the treatment of humans. Thus, pork meat may play a role in the transmission of antibiotic-resistant S. enterica subsp. arizonae to human consumers. This is the first report of S. enterica subsp. arizonae isolation from pigs.

  18. Isolamento de esporos de Paenibacillus larvae subsp. larvae no Brasil Detectionof Paenibacillus larvae subsp. larvae spores in Brazil

    Directory of Open Access Journals (Sweden)

    Dulce Maria Tocchetto Schuch

    2003-03-01

    Full Text Available Este trabalho objetivou detectar presença de esporos de Paenibacillus larvae subsp. larvae em produtos de um entreposto do interior do Estado do Rio Grande do Sul, a identificação de possíveis fontes de contaminação e a avaliação da possibilidade da transferência de esporos para colméias de apiários adjacentes a partir de produtos importados contaminados. Foram analisados mel e pólen importados disponíveis no entreposto, favo do ninho (crias, pólen e mel colhido de uma colméia sadia, mel estocado em um dos apiários e abelhas adultas. Os resultados foram positivosem relação ao mel e pólen importados, a três grupos de abelhas adultas e ao mel do favo.The objective of this work was to detect the presence of Paenibacillus larvae subsp. larvae spores in products from a warehouse located in Rio Grande do Sul State, Brazil, the identification of possible contamination sources, and the assessment of spores transference possibility from contaminated imported products from the warehouse to apiaries located in the surrounding area. Samples of imported pollen and bulk honey stocked in the warehouse, and honeycomb (brood, honey and pollen from a healthy hive, honey from one apiary and adult bees were analyzed. Imported honey and pollen, and three groups of adult bees and the honey collected from the honeycomb resulted positive.

  19. Protein Profile and Plasmid Content of Lactococcus lactis subsp. lactis LL52 and Lactococcus lactis subsp. cremoris LC79 Strains under Several Stress Conditions

    OpenAIRE

    LALE, Rahmi; TÜKEL, Çağla; AKÇELİK, Mustafa

    2014-01-01

    Differences in the protein and plasmid content of 2 Lactococcus lactis strains, L. lactis subsp. lactis LL52 and L. lactis subsp. cremoris LC79, under the stresses of high and low temperature, osmotic shock, and low pH were determined. We identified 3 new proteins with molecular masses of 16.0, 29.4, and 45.0 kDa as high temperature stress response specific in strain LL52. High temperature stress did not cause any changes in the protein content of strain LC79. Proteins that were specific for ...

  20. Dose-response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults

    DEFF Research Database (Denmark)

    Larsen, C.N.; Nielsen, S.; Kaestel, P.

    2006-01-01

    Objective: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora was analy......Objective: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora...... was analyzed in the 10(10) CFU/day probiotic and placebo group. Design: The study was designed as a randomized, placebo-controlled, double-blinded, parallel dose-response study. Subjects: Healthy young adults (18 - 40 years) were recruited by advertising in local newspapers. Of the 75 persons enrolled, 71 ( 46...... women, 25 men, mean age 25.6 years ( range 18 - 40 years)) completed the study. Intervention: The volunteers were randomly assigned into five groups receiving either placebo or a mixture of the two probiotics in the concentration of 10(8), 10(9), 10(10) or 10(11) CFU/day in 2 weeks run-in period, 3...

  1. Bacillus thuringiensis subsp. israelensis and Its Dipteran-Specific Toxins

    Science.gov (United States)

    Ben-Dov, Eitan

    2014-01-01

    Bacillus thuringiensis subsp. israelensis (Bti) is the first Bacillus thuringiensis to be found and used as an effective biological control agent against larvae of many mosquito and black fly species around the world. Its larvicidal activity resides in four major (of 134, 128, 72 and 27 kDa) and at least two minor (of 78 and 29 kDa) polypeptides encoded respectively by cry4Aa, cry4Ba, cry11Aa, cyt1Aa, cry10Aa and cyt2Ba, all mapped on the 128 kb plasmid known as pBtoxis. These six δ-endotoxins form a complex parasporal crystalline body with remarkably high, specific and different toxicities to Aedes, Culex and Anopheles larvae. Cry toxins are composed of three domains (perforating domain I and receptor binding II and III) and create cation-selective channels, whereas Cyts are composed of one domain that acts as well as a detergent-like membrane perforator. Despite the low toxicities of Cyt1Aa and Cyt2Ba alone against exposed larvae, they are highly synergistic with the Cry toxins and hence their combinations prevent emergence of resistance in the targets. The lack of significant levels of resistance in field mosquito populations treated for decades with Bti-bioinsecticide suggests that this bacterium will be an effective biocontrol agent for years to come. PMID:24686769

  2. Bacillus thuringiensis subsp. israelensis and Its Dipteran-Specific Toxins

    Directory of Open Access Journals (Sweden)

    Eitan Ben-Dov

    2014-03-01

    Full Text Available Bacillus thuringiensis subsp. israelensis (Bti is the first Bacillus thuringiensis to be found and used as an effective biological control agent against larvae of many mosquito and black fly species around the world. Its larvicidal activity resides in four major (of 134, 128, 72 and 27 kDa and at least two minor (of 78 and 29 kDa polypeptides encoded respectively by cry4Aa, cry4Ba, cry11Aa, cyt1Aa, cry10Aa and cyt2Ba, all mapped on the 128 kb plasmid known as pBtoxis. These six δ-endotoxins form a complex parasporal crystalline body with remarkably high, specific and different toxicities to Aedes, Culex and Anopheles larvae. Cry toxins are composed of three domains (perforating domain I and receptor binding II and III and create cation-selective channels, whereas Cyts are composed of one domain that acts as well as a detergent-like membrane perforator. Despite the low toxicities of Cyt1Aa and Cyt2Ba alone against exposed larvae, they are highly synergistic with the Cry toxins and hence their combinations prevent emergence of resistance in the targets. The lack of significant levels of resistance in field mosquito populations treated for decades with Bti-bioinsecticide suggests that this bacterium will be an effective biocontrol agent for years to come.

  3. Metabolic engineering of Propionibacterium freudenreichii subsp. shermanii for xylose fermentation.

    Science.gov (United States)

    Wei, Peilian; Lin, Meng; Wang, Zhongqiang; Fu, Hongxin; Yang, Hopen; Jiang, Wenyan; Yang, Shang-Tian

    2016-11-01

    Propionibacterium freudenreichii cannot use xylose, the second most abundant sugar in lignocellulosic biomass. Although Propionibacterium acidipropionici can use xylose as a carbon source, it is difficult to genetically modify, impeding further improvement through metabolic engineering. This study identified three xylose catabolic pathway genes encoding for xylose isomerase (xylA), xylose transporter (xylT), and xylulokinase (xylB) in P. acidipropionici and overexpressed them in P. freudenreichii subsp. shermanii via an expression plasmid pKHEM01, enabling the mutant to utilize xylose efficiently even in the presence of glucose without glucose-induced carbon catabolite repression. The mutant showed similar fermentation kinetics with glucose, xylose, and the mixture of glucose and xylose, respectively, as carbon source, and with or without the addition of antibiotic for selection pressure. The engineered P. shermanii thus can provide a novel cell factory for industrial production of propionic acid and other value-added products from lignocellulosic biomass. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Description of a Novel Adhesin of Mycobacterium avium Subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Mariana Noelia Viale

    2014-01-01

    Full Text Available The binding and ingestion of Mycobacterium avium subsp. paratuberculosis (MAP by host cells are fibronectin (FN dependent. In several species of mycobacteria, a specific family of proteins allows the attachment and internalization of these bacteria by epithelial cells through interaction with FN. Thus, the identification of adhesion molecules is essential to understand the pathogenesis of MAP. The aim of this study was to identify and characterize FN binding cell wall proteins of MAP. We searched for conserved adhesins within a large panel of surface immunogenic proteins of MAP and investigated a possible interaction with FN. For this purpose, a cell wall protein fraction was obtained and resolved by 2D electrophoresis. The immunoreactive spots were identified by MALDI-TOF MS and a homology search was performed. We selected elongation factor Tu (EF-Tu as candidate for further studies. We demonstrated the FN-binding capability of EF-Tu using a ligand blot assay and also confirmed the interaction with FN in a dose-dependent manner by ELISA. The dissociation constant of EF-Tu was determined by surface plasmon resonance and displayed values within the μM range. These data support the hypothesis that this protein could be involved in the interaction of MAP with epithelial cells through FN binding.

  5. Métodos de preservação de Acidovorax avenae subsp. citrulli Preservation of Acidovorax avenae subsp. citrulli

    Directory of Open Access Journals (Sweden)

    Dário Venâncio de Araújo

    2008-06-01

    Full Text Available Acidovorax avenae subsp. citrulli (Aac, agente da mancha-aquosa, causa grandes prejuízos ao melão e outras cucurbitáceas no Brasil e no mundo. Os métodos dessecação em papel de filtro, repicagens periódicas, água esterilizada e folhas herborizadas foram testados para preservação de Aac1 e Aac1.12 durante 180 dias. Mensalmente, a viabilidade de Aac foi avaliada pelo crescimento em meio de cultura e a patogenicidade das culturas viáveis foi avaliada pela incidência e severidade da doença em plântulas de melão. A preservação em papel de filtro resultou em 100% de viabilidade dos isolados durante o período, enquanto que nos demais métodos houve perda de viabilidade no decorrer das avaliações. Os métodos de dessecação em papel de filtro e o de repicagens periódicas foram mais eficientes que a água esterilizada e folhas herborizadas na manutenção da patogenicidade dos isolados durante os 180 dias.The phytopathogenic bacteria Acidovorax avenae subsp. citrulli (Aac, agent of bacterial blotch, causes severe damages to melon and other cucurbits in Brazil and worlwide. The methods desiccation in filter paper, periodic transfer, sterile water and dried leaves were tested for preserving the strains Aac1 and Aac1.12 of this bacterium during 180 days. Evaluations of bacterial viability were performed monthly by growing strains on culture media. The pathogenicity of viable cultures was evaluated by disease incidence and severity on melon seedlings. The desiccation in filter paper maintained 100% viability of the strains during the period while using the other methods, viability was lost during evaluations. Desiccation in filter paper and periodic transfer were more efficient than sterile water and dried leaves in maintening strain pathogenicity during the time evaluated 180 days.

  6. Novel Streptococcus infantarius subsp. infantarius variants harboring lactose metabolism genes homologous to Streptococcus thermophilus.

    Science.gov (United States)

    Jans, Christoph; Gerber, Andrea; Bugnard, Joséphine; Njage, Patrick Murigu Kamau; Lacroix, Christophe; Meile, Leo

    2012-08-01

    Streptococcus infantarius subsp. infantarius belongs to the Streptococcus bovis/Streptococcus equinus complex (SBSEC) commonly associated with human and animal infections. We elucidated the lactose metabolism of S. infantarius subsp. infantarius predominant in African fermented milk products. S. infantarius subsp. infantarius isolates (n = 192) were identified in 88% of spontaneously fermented camel milk suusac samples (n = 24) from Kenya and Somalia at log₁₀ 8.2-8.5 CFU mL⁻¹. African S. infantarius isolates excreted stoichiometric amounts of galactose when grown on lactose, exhibiting a metabolism similar to Streptococcus thermophilus and distinct from their type strain. African S. infantarius subsp. infantarius CJ18 harbors a regular gal operon with 99.7-100% sequence identity to S. infantarius subsp. infantarius ATCC BAA-102(T) and a gal-lac operon with 91.7-97.6% sequence identity to S. thermophilus, absent in all sequenced SBSEC strains analyzed. The expression and functionality of lacZ was demonstrated in a β-galactosidase assay. The gal-lac operon was identified in 100% of investigated S. infantarius isolates (n = 46) from suusac samples and confirmed in Malian fermented cow milk isolates. The African S. infantarius variant potentially evolved through horizontal gene transfer of an S. thermophilus-homologous lactose pathway. Safety assessments are needed to identify any putative health risks of this novel S. infantarius variant. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Persistence of Bacillus thuringiensis subsp. kurstaki in Urban Environments following Spraying▿†‡

    Science.gov (United States)

    Van Cuyk, Sheila; Deshpande, Alina; Hollander, Attelia; Duval, Nathan; Ticknor, Lawrence; Layshock, Julie; Gallegos-Graves, LaVerne; Omberg, Kristin M.

    2011-01-01

    Bacillus thuringiensis subsp. kurstaki is applied extensively in North America to control the gypsy moth, Lymantria dispar. Since B. thuringiensis subsp. kurstaki shares many physical and biological properties with Bacillus anthracis, it is a reasonable surrogate for biodefense studies. A key question in biodefense is how long a biothreat agent will persist in the environment. There is some information in the literature on the persistence of Bacillus anthracis in laboratories and historical testing areas and for Bacillus thuringiensis in agricultural settings, but there is no information on the persistence of Bacillus spp. in the type of environment that would be encountered in a city or on a military installation. Since it is not feasible to release B. anthracis in a developed area, the controlled release of B. thuringiensis subsp. kurstaki for pest control was used to gain insight into the potential persistence of Bacillus spp. in outdoor urban environments. Persistence was evaluated in two locations: Fairfax County, VA, and Seattle, WA. Environmental samples were collected from multiple matrices and evaluated for the presence of viable B. thuringiensis subsp. kurstaki at times ranging from less than 1 day to 4 years after spraying. Real-time PCR and culture were used for analysis. B. thuringiensis subsp. kurstaki was found to persist in urban environments for at least 4 years. It was most frequently detected in soils and less frequently detected in wipes, grass, foliage, and water. The collective results indicate that certain species of Bacillus may persist for years following their dispersal in urban environments. PMID:21926205

  8. Diverse Antibacterial activity of Pectobacterium carotovorum subsp.carotovorum isolated in Korea.

    Science.gov (United States)

    Roh, Eunjung; Lee, Seungdon; Lee, Yonghoon; Ra, Dongsu; Choi, Jaehyuk; Moon, Eunpyo; Heu, Sunggi

    2009-01-01

    Fifty-four Pectobacterium carotovorum subsp. carotovorum strains isolated in Korea were characterized by a spectrum of antibacterial activities against 7 indicator strains chosen to represent various regions and host plants. All P. carotovorum subsp. carotovorum isolates tested could be grouped into 4 classes depending on the pattern of antibacterial substance production. All tested strains had DNA fragment(s) homologous to the genes encoding carotovoricin and 21 of them had genes homologous to DNA invertase. Sixteen strains had genes homologous to the genes encoding carocin S1. Several isolates produced antibacterial substances active against strains in Brenneria, Pantoea, and Pectobacterium genera that belonged formerly to the genus Erwinia. Strains in Pseudomonas or Xanthomonas sp. were not sensitive to the antibacterial substances produced by P. carotovorum subsp. carotovorum, except for X. albilineans that was sensitive to antibacterial substances produced by most strains in P. carotovorum subsp. carotovorum and P. betavasculorum KACC10056. These results demonstrated the diverse patterns of antibacterial substance production and the possibility of the existence of new antibacterial substance(s) produced by P. carotovorum subsp. carotovorum isolated in Korea.

  9. Xylella fastidiosa Isolates from Both subsp. multiplex and fastidiosa Cause Disease on Southern Highbush Blueberry (Vaccinium sp.) Under Greenhouse Conditions.

    Science.gov (United States)

    Oliver, J E; Cobine, P A; De La Fuente, L

    2015-07-01

    Xylella fastidiosa is a xylem-limited gram-negative plant pathogen that affects numerous crop species, including grape, citrus, peach, pecan, and almond. Recently, X. fastidiosa has also been found to be the cause of bacterial leaf scorch on blueberry in the southeastern United States. Thus far, all X. fastidiosa isolates obtained from infected blueberry have been classified as X. fastidiosa subsp. multiplex; however, X. fastidiosa subsp. fastidiosa isolates are also present in the southeastern United States and commonly cause Pierce's disease of grapevines. In this study, seven southeastern U.S. isolates of X. fastidiosa, including three X. fastidiosa subsp. fastidiosa isolates from grape, one X. fastidiosa subsp. fastidiosa isolate from elderberry, and three X. fastidiosa subsp. multiplex isolates from blueberry, were used to infect the southern highbush blueberry 'Rebel'. Following inoculation, all isolates colonized blueberry, and isolates from both X. fastidiosa subsp. multiplex and X. fastidiosa subsp. fastidiosa caused symptoms, including characteristic stem yellowing and leaf scorch symptoms as well as dieback of the stem tips. Two X. fastidiosa subsp. multiplex isolates from blueberry caused more severe symptoms than the other isolates examined, and infection with these two isolates also had a significant impact on host mineral nutrient content in sap and leaves. These findings have potential implications for understanding X. fastidiosa host adaptation and expansion and the development of emerging diseases caused by this bacterium.

  10. The morphological and anatomical studies on endemic crocus biflorus miller subsp. Pulchricolor (herbert) mathew (iridaceae) in turkey

    International Nuclear Information System (INIS)

    Akyol, Y.

    2014-01-01

    In this study, the morphological and anatomical characteristics of Crocus biflorus subsp. pulchricolor (Iridaceae)were investigated. The subsp. pulchricolor has, 4 leaves, 1 mm broad, bracts drying brownish. these properties are characteristics of these plants. In anatomical studies, cross-sections of the root, stem and leaves were examined. These parts photographed and compared with the other crocus and Iridaceae family species. (author)

  11. Dissemination of the superantigen encoding genes seeL, seeM, szeL and szeM in Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus.

    Science.gov (United States)

    Alber, J; El-Sayed, A; Estoepangestie, S; Lämmler, C; Zschöck, M

    2005-08-10

    Bacterial superantigens are one of the major virulence factors produced by Streptococcus pyogenes and Staphylococcus aureus. The two novel superantigen encoding genes seeM and seeL were described for S. equi subsp. equi which is known as the causative agent of strangles in equids. In the present study previously characterized S. equi subsp. equi strains and strains of various other animal pathogenic streptococcal species and subspecies were investigated for the presence of the superantigen encoding genes seeM and seeL by polymerase chain reaction. According to these studies seeL and seeM appeared to be a constant characteristic of all investigated S. equi subsp. equi strains. Surprisingly, one S. equi subsp. zooepidemicus strain (S.z. 122) was also positive for both genes. The species identity of this S. equi subsp. zooepidemicus strain could additionally be confirmed by sequencing the 16S rRNA gene and the 16S-23S rDNA intergenic spacer region. The superantigen encoding genes could not be found among additionally investigated S. equi subsp. zooepidemicus strains or among strains of seven other streptococcal species. The seeL and seeM genes of the S. equi subsp. equi strain S.e. CF32 and the genes szeL and szeM of the S. equi subsp. zooepidemicus strain S.z. 122 were cloned and sequenced. A sequence comparison revealed a high degree of sequence homology between seeL, szeL, speL and seeM, szeM and speM, respectively. The superantigenic toxins L and M seemed to be widely distributed virulence factors of S. equi subsp. equi, rare among S. equi subsp. zooepidemicus but did not occur among a number of other animal pathogenic streptococcal species.

  12. Chemical composition of essential oil of mentha longifolia l. subsp. longifolia growing wild

    International Nuclear Information System (INIS)

    Okut, N.; Yagmur, M.; Yildirim, B.

    2017-01-01

    The essential oil of Mentha longifolia L., is very important to some culinary usage and antimicrobial activity. The essential oil of Mentha longifolia subsp. longifolia growing in the Bahcesaray area (Van Province, Turkey) was studied. This study designed for determine of essential oil constituent Mentha longifolia subsp. longifolia that collected from wild area. Mint leaves sample essential oils obtained by hydro distillation and essential oil components were determined using GC-MS. The main component of wild grown Mentha longifolia subsp. longifolia was Menthone (19.31%). Second one and others were Pulegone (12.42%), Piperitone (11.05%), Dihydrocarvon (8.32%), Limonene (6.1%), 3-Terpinolenone (5.66%), 1,8-Cineole (4.37%), Germacrene D (3.38%) and Caryopyllene (3.19%), respectively. (author)

  13. Complete genome sequence of phytopathogenic Pectobacterium carotovorum subsp. carotovorum bacteriophage PP1.

    Science.gov (United States)

    Lee, Ju-Hoon; Shin, Hakdong; Ji, Samnyu; Malhotra, Shweta; Kumar, Mukesh; Ryu, Sangryeol; Heu, Sunggi

    2012-08-01

    Pectobacterium carotovorum subsp. carotovorum is a phytopathogen causing soft rot disease on diverse plant species. To control this plant pathogen, P. carotovorum subsp. carotovorum-targeting bacteriophage PP1 was isolated and its genome was completely sequenced to develop a novel biocontrol agent. Interestingly, the 44,400-bp genome sequence does not encode any gene involved in the formation of lysogen, suggesting that this phage may be very useful as a biocontrol agent because it does not make lysogen after host infection. This is the first report on the complete genome sequence of the P. carotovorum subsp. carotovorum-targeting bacteriophage, and it will enhance our understanding of the interaction between phytopathogens and their targeting bacteriophages.

  14. [Function of flgK gene in Pectobacterium carotovorum subsp, carotovorum].

    Science.gov (United States)

    Yang, Zhongling; Deng, Yamin; Du, Shuo; Li, Ting; Jiang, Huan; Liu, Fengquan; Fan, Jiaqin

    2012-06-04

    To investigate functions of flgKpcc gene in Pectobacterium carotovorum subsp. carotovorum (P. c. c). The gene knock-out mutant deltaflgKpcc and complemented strain deltaflgKpcc-KH were generated by biparental mating and their phenotypes including cell morphology, motility, pathogenic factors, and pathogenicity were investigated. Non-flagellum, cell precipitation in the culture and significantly attenuated motility on 0.3% semisolid medium were observed in deltaflgKpcc compared to Pectobacterium carotovorum subsp. carotovorum S1. In addition, significant decrease in cellulase and protease activity, biofilm formation, and pathogenicity on host plant were found in deltaflgKpcc. While there were no apparent difference in growth rate in vitro, deltaflgKpcc-KH, the complementation strain, restored the phenotype of deltaflgKpcc to the wild type level. The gen of flgKpcc not only influences the cell motility, but also pathogenic factors to lead to the decreased pathogenicity in Pectobacterium carotovorum subsp. Carotovorum.

  15. Comparative Phenotypic and Molecular Genetic Profiling of Wild Lactococcus lactis subsp. lactis Strains of the L. lactis subsp. lactis and L. lactis subsp. cremoris Genotypes, Isolated from Starter-Free Cheeses Made of Raw Milk▿

    Science.gov (United States)

    Fernández, Elena; Alegría, Ángel; Delgado, Susana; Martín, M. Cruz; Mayo, Baltasar

    2011-01-01

    Twenty Lactococcus lactis strains with an L. lactis subsp. lactis phenotype isolated from five traditional cheeses made of raw milk with no added starters belonging to the L. lactis subsp. lactis and L. lactis subsp. cremoris genotypes (lactis and cremoris genotypes, respectively; 10 strains each) were subjected to a series of phenotypic and genetic typing methods, with the aims of determining their phylogenetic relationships and suitability as starters. Pulsed-field gel electrophoresis (PFGE) analysis of intact genomes digested with SalI and SmaI proved that all strains were different except for three isolates of the cremoris genotype, which showed identical PFGE profiles. Multilocus sequence typing (MLST) analysis using internal sequences of seven loci (namely, atpA, rpoA, pheS, pepN, bcaT, pepX, and 16S rRNA gene) revealed considerable intergenotype nucleotide polymorphism, although deduced amino acid changes were scarce. Analysis of the MLST data for the present strains and others from other dairy and nondairy sources showed that all of them clustered into the cremoris or lactis genotype group, by using both independent and combined gene sequences. These two groups of strains also showed distinctive carbohydrate fermentation and enzyme activity profiles, with the strains in the cremoris group showing broader profiles. However, the profiles of resistance/susceptibility to 16 antibiotics were very similar, showing no atypical resistance, except for tetracycline resistance in three identical cremoris genotype isolates. The numbers and concentrations of volatile compounds produced in milk by the strains belonging to these two groups were clearly different, with the cremoris genotype strains producing higher concentrations of more branched-chain, derived compounds. Together, the present results support the idea that the lactis and cremoris genotypes of phenotypic Lactococcus lactis subsp. lactis actually represent true subspecies. Some strains of the two subspecies

  16. Characterization of a novel bacteriophage, Phda1, infecting the histamine-producing Photobacterium damselae subsp. damselae.

    Science.gov (United States)

    Yamaki, S; Kawai, Y; Yamazaki, K

    2015-06-01

    Photobacterium damselae subsp. damselae is a potent histamine-producing micro-organism. The aim of this study was to isolate and characterize a bacteriophage Phda1 that infected P. damselae subsp. damselae to inhibit its growth and histamine accumulation. Phda1 was isolated from a raw oyster, and the host range, morphology and the bacteriophage genome size were analysed. Phda1 formed a clear plaque only against P. damselae subsp. damselae JCM8969 among five Gram-positive and 32 Gram-negative bacterial strains tested. Phda1 belongs to the family Myoviridae, and its genome size was estimated as 35·2-39·5 kb. According to the one-step growth curve analysis, the latent period, rise period and burst size of Phda1 were 60 min, 50 min and 19 plaque-forming units per infected cell, respectively. Divalent cations, especially Ca(2+) and Mg(2+) , strongly improved Phda1 adsorption to the host cells and its propagation. Phda1 treatment delayed the growth and histamine production of P. damselae subsp. damselae in an in vitro challenge test. The bacteriophage Phda1 might serve as a potential antimicrobial agent to inhibit the histamine poisoning caused by P. damselae subsp. damselae. This is the first description of a bacteriophage specifically infecting P. damselae subsp. damselae and its potential applications. Bacteriophage therapy could prove useful in the prevention of histamine poisoning. © 2015 The Society for Applied Microbiology.

  17. Microbiota of Minas cheese as influenced by the nisin producer Lactococcus lactis subsp. lactis GLc05.

    Science.gov (United States)

    Perin, Luana Martins; Dal Bello, Barbara; Belviso, Simona; Zeppa, Giuseppe; Carvalho, Antônio Fernandes de; Cocolin, Luca; Nero, Luís Augusto

    2015-12-02

    Minas cheese is a popular dairy product in Brazil that is traditionally produced using raw or pasteurized cow milk. This study proposed an alternative production of Minas cheese using raw goat milk added of a nisin producer Lactococcus lactis subsp. lactis GLc05. An in situ investigation was carried on to evaluate the interactions between the L. lactis subsp. lactis GLc05 and the autochthonous microbiota of a Minas cheese during the ripening; production of biogenic amines (BAs) was assessed as a safety aspect. Minas cheese was produced in two treatments (A, by adding L. lactis subsp. lactis GLc05, and B, without adding this strain), in three independent repetitions (R1, R2, and R3). Culture dependent (direct plating) and independent (rep-PCR and PCR-DGGE) methods were employed to characterize the microbiota and to assess the possible interferences caused by L. lactis subsp. lactis GLc05. BA amounts were measured using HPLC. A significant decrease in coagulase-positive cocci was observed in the cheeses produced by adding L. lactis subsp. lactis GLc05 (cheese A). The rep-PCR and PCR-DGGE highlighted the differences in the microbiota of both cheeses, separating them into two different clusters. Lactococcus sp. was found as the main microorganism in both cheeses, and the microbiota of cheese A presented a higher number of species. High concentrations of tyramine were found in both cheeses and, at specific ripening times, the BA amounts in cheese B were significantly higher than in cheese A (pcheese and by controlling the growth of coagulase-positive cocci. L. lactis subsp. lactis GLc05 influenced also the production of BA determining that their amounts in the cheeses were maintained at acceptable levels for human consumption. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Interaction between Mycobacterium avium subsp. paratuberculosis and environmental protozoa

    Directory of Open Access Journals (Sweden)

    Rowe Michael T

    2006-07-01

    Full Text Available Abstract Background Interactions between Mycobacterium avium subsp. paratuberculosis (Map and free-living protozoa in water are likely to occur in nature. The potential impact of ingestion of Map by two naturally occurring Acanthamoeba spp. on this pathogen's survival and chlorine resistance was investigated. Results Between 4.6 and 9.1% of spiked populations of three Map strains (NCTC 8578, B2 and ATCC 19698, which had been added at a multiplicity of infection of 10:1, were ingested by Acanthamoeba castellanii CCAP 1501/1B and A. polyphaga CCAP 1501/3B during co-culture for 3 h at 25°C. Map cells were observed to be present within the vacuoles of the amoebae by acid-fast staining. During extended co-culture of Map NCTC 8578 at 25°C for 24 d with both A. castellanii and A. polyphaga Map numbers did not change significantly during the first 7 days of incubation, however a 1–1.5 log10 increase in Map numbers was observed between days 7 and 24 within both Acanthamoeba spp. Ingested Map cells were shown to be more resistant to chlorine inactivation than free Map. Exposure to 2 μg/ml chlorine for 30 min resulted in a log10 reduction of 0.94 in ingested Map but a log10 reduction of 1.73 in free Map (p Conclusion This study demonstrated that ingestion of Map by and survival and multiplication of Map within Acanthamoeba spp. is possible, and that Map cells ingested by amoebae are more resistant to inactivation by chlorine than free Map cells. These findings have implications with respect to the efficacy of chlorination applied to Map infected surface waters.

  19. Molecular Subtyping of Treponema pallidum subsp. pallidum in Lisbon, Portugal▿

    Science.gov (United States)

    Castro, R.; Prieto, E.; Águas, M. J.; Manata, M. J.; Botas, J.; Martins Pereira, F.

    2009-01-01

    The objectives of this study were to evaluate the reproducibility of a molecular method for the subtyping of Treponema pallidum subsp. pallidum and to discriminate strains of this microorganism from strains from patients with syphilis. We studied 212 specimens from a total of 82 patients with different stages of syphilis (14 primary, 7 secondary and 61 latent syphilis). The specimens were distributed as follows: genital ulcers (n = 9), skin and mucosal lesions (n = 7), blood (n = 82), plasma (n = 82), and ear lobe scrapings (n = 32). The samples were assayed by a PCR technique to amplify a segment of the polymerase gene I (polA). Positive samples were typed on the basis of the analysis of two variable genes, tpr and arp. Sixty-two of the 90 samples positive for polA yielded typeable Treponema pallidum DNA. All skin lesions in which T. pallidum was identified (six of six [100%]) were found to contain enough DNA for typing of the organism. It was also possible to type DNA from 7/9 (77.7%) genital ulcer samples, 13/22 (59.1%) blood samples, 20/32 (62.5%) plasma samples, and 16/21 (76.2%) ear lobe scrapings. The same subtype was identified in all samples from the same patient. Five molecular subtypes (subtypes 10a, 14a, 14c, 14f, and 14g) were identified, with the most frequently found subtype being subtype 14a and the least frequently found subtype being subtype 10a. In conclusion, the subtyping technique used in this study seems to have good reproducibility. To our knowledge, subtype 10a was identified for the first time. Further studies are needed to explain the presence of this subtype in Portugal, namely, its relationship to the Treponema pallidum strains circulating in the African countries where Portuguese is spoken. PMID:19494073

  20. Characterization of free exopolysaccharides secreted by Mycoplasma mycoides subsp. mycoides.

    Directory of Open Access Journals (Sweden)

    Clothilde Bertin

    Full Text Available Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm. In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic functions and specific components of the outer cell surface. One of these latter, the capsular polysaccharide galactan has been notably demonstrated to play a role in Mmm persistence and dissemination. The free exopolysaccharides (EPS, also produced by Mmm and shown to circulate in the blood stream of infected cattle, have received little attention so far. Indeed, their characterization has been hindered by the presence of polysaccharide contaminants in the complex mycoplasma culture medium. In this study, we developed a method to produce large quantities of EPS by transfer of mycoplasma cells from their complex broth to a chemically defined medium and subsequent purification. NMR analyses revealed that the purified, free EPS had an identical β(1->6-galactofuranosyl structure to that of capsular galactan. We then analyzed intraclonal Mmm variants that produce opaque/translucent colonies on agar. First, we demonstrated that colony opacity was related to the production of a capsule, as observed by electron microscopy. We then compared the EPS extracts and showed that the non-capsulated, translucent colony variants produced higher amounts of free EPS than the capsulated, opaque colony variants. This phenotypic variation was associated with an antigenic variation of a specific glucose phosphotransferase permease. Finally, we conducted in silico analyses of candidate polysaccharide biosynthetic pathways in order to decipher the potential link between glucose phosphotransferase permease activity and attachment/release of galactan. The co-existence of variants producing alternative forms of galactan (capsular versus free extracellular galactan and associated

  1. Virulence differences among Francisella tularensis subsp. tularensis clades in mice.

    Directory of Open Access Journals (Sweden)

    Claudia R Molins

    Full Text Available Francisella tularensis subspecies tularensis (type A and holarctica (type B are of clinical importance in causing tularemia. Molecular typing methods have further separated type A strains into three genetically distinct clades, A1a, A1b and A2. Epidemiological analyses of human infections in the United States suggest that A1b infections are associated with a significantly higher mortality rate as compared to infections caused by A1a, A2 and type B. To determine if genetic differences as defined by molecular typing directly correlate with differences in virulence, A1a, A1b, A2 and type B strains were compared in C57BL/6 mice. Here we demonstrate significant differences between survival curves for infections caused by A1b versus A1a, A2 and type B, with A1b infected mice dying earlier than mice infected with A1a, A2 or type B; these results were conserved among multiple strains. Differences were also detected among type A clades as well as between type A clades and type B with respect to bacterial burdens, and gross anatomy in infected mice. Our results indicate that clades defined within F. tularensis subsp. tularensis by molecular typing methods correlate with virulence differences, with A1b strains more virulent than A1a, A2 and type B strains. These findings indicate type A strains are not equivalent with respect to virulence and have important implications for public health as well as basic research programs.

  2. Is the Evolution of Salmonella enterica subsp. enterica Linked to Restriction-Modification Systems?

    DEFF Research Database (Denmark)

    Roer, Louise; Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas

    2016-01-01

    Salmonella enterica subsp. enterica bacteria are highly diverse foodborne pathogens that are subdivided into more than 1,500 serovars. The diversity is believed to result from mutational evolution, as well as intra- and interspecies recombination that potentially could be influenced by restrictio...... to the conjugational mode of horizontal gene transfer in Salmonella. Thus, we conclude that other factors must be involved in shaping the evolution of bacteria.......Salmonella enterica subsp. enterica bacteria are highly diverse foodborne pathogens that are subdivided into more than 1,500 serovars. The diversity is believed to result from mutational evolution, as well as intra- and interspecies recombination that potentially could be influenced by restriction...

  3. Characteristics of the Leuconostoc mesenteroides subsp. mesenteroides strains from fresh vegetables

    Directory of Open Access Journals (Sweden)

    Dimić Gordana R.

    2006-01-01

    Full Text Available Strains synthesizing extracellular polysaccharide dextran on a medium with 10% sucrose were isolated from different kind of vegetables (cabbage, cucumber, cauliflower, kohlrabi, carrot, green beans, red beet, pepper, eggplant, radish. Carbohydrate fermentation was examined using a bioMerieux API 50 CHL test system. Among micropopulations with characteristic spherical cell morphology, 94.9% belonged to Leuconostoc mesenteroides subsp. mesenteroides and 5.1% were identified as Leuconostoc mesenteroides subsp. dextranicum. According to fermentation of pentoses L. mesenteroides strains were divided into three groups with a certain number of biotypes; 10 strains were tested on acid production. .

  4. Determination of antioxidant properties of Rumex crispus and Scrophularia canina subsp. bicolor

    OpenAIRE

    Demir, Serhat; Bozkurt, Buket; Önür, Mustafa Ali; Kaya, İrem Gülen; Ünver Somer, Nehir

    2017-01-01

    Methanoland ethyl acetate extracts of Rumex crispus L. and Scrophularia canina L. subsp. bicolor (SM.) Greuter were tested fortheir antioxidant activity using the DPPH method. Extracts were prepared fromthe above-ground parts of these plants. Significant antioxidant activity wasdetermined for methanol (IC50: 4.16 µg/mL) and ethyl acetate (IC50:8.71 µg/mL) extracts of Rumex crispus.Moreover, methanol (IC50: 60.78 µg/mL) and ethyl acetate (IC50:149.33 µg/mL) extracts of Scrophulariacanina subsp...

  5. Essential oil composition of the fruits of Periploca laevigata Aiton subsp. angustifolia (Labill.) Markgraf (Apocynaceae - Periplocoideae).

    Science.gov (United States)

    Zito, Pietro; Sajeva, Maurizio; Bruno, Maurizio; Maggio, Antonella; Rosselli, Sergio; Senatore, Felice; Formisano, Carmen

    2011-08-01

    The essential oil of the fruits of Periploca laevigata Aiton subsp. angustifolia (Labill.) Markgraf (Apocynaceae) from Lampedusa Island was obtained by hydrodistillation and its composition was analysed. The analyses allowed the identification and quantification of 64 volatile compounds belonging to different classes. The most abundant compounds were nonacosane, heptacosane, hentriacontane and δ-cadinene. Among the volatile compounds identified in the fruits of P. laevigata subsp. angustifolia, 31 are present in other taxa of Apocynaceae, 19 have antimicrobial activity and four are pheromones for the butterfly Danaus chrysippus. The possible ecological role of the volatile compounds found is briefly discussed.

  6. Screening for excellent mutants of Streptomyces roseoflavus Subsp. by N+ implantation

    International Nuclear Information System (INIS)

    Shi Yuefeng; Sang Jinlong; Zhu Lihong; Wu Wenjuan; Yao Hangli; Xia Zhanen; Huang Wencai

    2002-01-01

    The biological effects of Streptomyces roseoflavus subsp, hangzhouensisn. Subsp. with N + implantation are reported. The total and positive mutation rates with various doses (3 x 10 15 - 15 x 10 15 N + /cm 2 ) were 42.4% - 73.0% and 5.8% -38.2%, respectively. Mutant 97 - 49 with potency over four times more than that of the parental strain was obtained through N + implantation and showed good genetic stability. These results showed that ion implantation was an effective method for microbe mutagenesis

  7. Complete genome sequence of Pectobacterium carotovorum subsp. carotovorum bacteriophage My1.

    Science.gov (United States)

    Lee, Dong Hwan; Lee, Ju-Hoon; Shin, Hakdong; Ji, Samnyu; Roh, Eunjung; Jung, Kyusuk; Ryu, Sangryeol; Choi, Jaehyuk; Heu, Sunggi

    2012-10-01

    Pectobacterium carotovorum subsp. carotovorum, a member of the Enterobacteriaceae family, is an important plant-pathogenic bacterium causing significant economic losses worldwide. P. carotovorum subsp. carotovorum bacteriophage My1 was isolated from a soil sample. Its genome was completely sequenced and analyzed for the development of an effective biological control agent. Sequence and morphological analyses revealed that phage My1 is a T5-like bacteriophage and belongs to the family Siphoviridae. To date, there is no report of a Pectobacterium-targeting siphovirus genome sequence. Here, we announce the complete genome sequence of phage My1 and report the results of our analysis.

  8. Isolation of Salmonella enterica subsp. enterica (O:4,5:i and Salmonella enterica subsp. Typhimurium from free-living domestic pigeons (Columba livia

    Directory of Open Access Journals (Sweden)

    R.C. Rocha-e-Silva

    2014-10-01

    Full Text Available The present study reports the isolation of Salmonella enterica in organs of free-living domestic pigeons. In the clinic examination, the presence of feces in the peri-cloacal and abdominal regions were observed, as well as symptoms such as cachexy, incoordination and opisthotonos. Before any therapeutic protocol was applied the bird died and a necropsy was then performed for the removal of spleen, liver, kidney and intestine for bacteriological examination and antibiotic sensitivity test. Salmonella enterica subsp.enterica (O:4,5:i- and Salmonella enterica subsp. enterica serovar Typhimurium were isolated from the liver and intestine and the sensitivity test demonstrated that these strains are sensitive to several antibiotics.

  9. Verminephrobacter aporrectodeae sp. nov. subsp. tuberculatae and subsp. caliginosae; the specific nephridial symbionts of the earthworms Aporrectodea tuberculata and A. caliginosa

    DEFF Research Database (Denmark)

    Lund, Marie Braad; Schätzle, Sarah; Schramm, Andreas

    2012-01-01

    .3%, their earthworm host specificity, differing temperature ranges and pH optima suggest that they represent two subspecies of a novel species of Verminephrobacter. For this species, the name V. aporrectodeae sp. nov. is proposed, with the two subspecies V. aporrectodeae subsp. tuberculatae (type strain, At4T = DSM...... 21361T = LMG 25313T) and V. aporrectodeae subsp. caliginosae (type strain, Ac9T = DSM 21895T = LMG 5312T) isolated from the nephridia of the earthworms Aporrectodea tuberculata and A. caliginosa, respectively....... analysis. Similarly to V. eiseniae, the two isolates grew aerobically with a preference for low oxygen concentrations on a range of sugars, fatty acids and amino acids and fermentatively on glucose and pyruvate. These phenotypes match well with the conditions reported or inferred for the nephridial...

  10. Complete Genome and Methylome Sequences of Salmonella enterica subsp. enterica Serovar Panama (ATCC 7378) and Salmonella enterica subsp. enterica Serovar Sloterdijk (ATCC 15791)

    OpenAIRE

    Yao, Kuan; Muruvanda, Tim; Roberts, Richard J.; Payne, Justin; Allard, Marc W.; Hoffmann, Maria

    2016-01-01

    Salmonella enterica spp. are pathogenic bacteria commonly associated with food-borne outbreaks in human and animals. Salmonella enterica spp. are characterized into more than 2,500 different serotypes, which makes epidemiological surveillance and outbreak control more difficult. In this report, we announce the first complete genome and methylome sequences from two Salmonella type strains associated with food-borne outbreaks, Salmonella enterica subsp. enterica serovar Panama (ATCC 7378) and S...

  11. Polyphasic taxonomic revision of the Ralstonia solanacearum species complex: proposal to emend the descriptions of Ralstonia solanacearum and Ralstonia syzygii and reclassify current R. syzygii strains as Ralstonia syzygii subsp. syzygii subsp. nov., R. solanacearum phylotype IV strains as Ralstonia syzygii subsp. indonesiensis subsp. nov., banana blood disease bacterium strains as Ralstonia syzygii subsp. celebesensis subsp. nov. and R. solanacearum phylotype I and III strains as Ralstonia pseudosolanacearum sp. nov.

    Science.gov (United States)

    Safni, Irda; Cleenwerck, Ilse; De Vos, Paul; Fegan, Mark; Sly, Lindsay; Kappler, Ulrike

    2014-09-01

    The Ralstonia solanacearum species complex has long been recognized as a group of phenotypically diverse strains that can be subdivided into four phylotypes. Using a polyphasic taxonomic approach on an extensive set of strains, this study provides evidence for a taxonomic and nomenclatural revision of members of this complex. Data obtained from phylogenetic analysis of 16S-23S rRNA ITS gene sequences, 16S-23S rRNA intergenic spacer (ITS) region sequences and partial endoglucanase (egl) gene sequences and DNA-DNA hybridizations demonstrate that the R. solanacearum species complex comprises three genospecies. One of these includes the type strain of Ralstonia solanacearum and consists of strains of R. solanacearum phylotype II only. The second genospecies includes the type strain of Ralstonia syzygii and contains only phylotype IV strains. This genospecies is subdivided into three distinct groups, namely R. syzygii, the causal agent of Sumatra disease on clove trees in Indonesia, R. solanacearum phylotype IV strains isolated from different host plants mostly from Indonesia, and strains of the blood disease bacterium (BDB), the causal agent of the banana blood disease, a bacterial wilt disease in Indonesia that affects bananas and plantains. The last genospecies is composed of R. solanacearum strains that belong to phylotypes I and III. As these genospecies are also supported by phenotypic data that allow the differentiation of the three genospecies, the following taxonomic proposals are made: emendation of the descriptions of Ralstonia solanacearum and Ralstonia syzygii and descriptions of Ralstonia syzygii subsp. nov. (type strain R 001(T) = LMG 10661(T) = DSM 7385(T)) for the current R. syzygii strains, Ralstonia syzygii subsp. indonesiensis subsp. nov. (type strain UQRS 464(T) = LMG 27703(T) = DSM 27478(T)) for the current R. solanacearum phylotype IV strains, Ralstonia syzygii subsp. celebesensis subsp. nov. (type strain UQRS 627(T

  12. Bioprocessing of some agro-industrial residues for endoglucanase production by the new subsp.; Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J.

    Science.gov (United States)

    El-Naggar, Noura El-Ahmady; Abdelwahed, Nayera A M; Saber, Wesam I A; Mohamed, Asem A

    2014-01-01

    The use of low cost agro-industrial residues for the production of industrial enzymes is one of the ways to reduce significantly production costs. Cellulase producing actinomycetes were isolated from soil and decayed agricultural wastes. Among them, a potential culture, strain NEAE-J, was selected and identified on the basis of morphological, cultural, physiological and chemotaxonomic properties, together with 16S rDNA sequence. It is proposed that strain NEAE-J should be included in the species Streptomyces albogriseolus as a representative of a novel sub-species, Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J and sequencing product was deposited in the GenBank database under accession number JN229412. This organism was tested for its ability to produce endoglucanase and release reducing sugars from agro-industrial residues as substrates. Sugarcane bagasse was the most suitable substrate for endoglucanase production. Effects of process variables, namely incubation time, temperature, initial pH and nitrogen source on production of endoglucanase by submerged fermentation using Streptomyces albogriseolus subsp. cellulolyticus have been studied. Accordingly optimum conditions have been determined. Incubation temperature of 30 °C after 6 days, pH of 6.5, 1% sugarcane bagasse as carbon source and peptone as nitrogen source were found to be the optimum for endoglucanase production. Optimization of the process parameters resulted in about 2.6 fold increase in the endoglucanase activity. Therefore, Streptomyces albogriseolus subsp. cellulolyticus coud be potential microorganism for the intended application.

  13. Bioprocessing of some agro-industrial residues for endoglucanase production by the new subsp.; Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J

    Directory of Open Access Journals (Sweden)

    Noura El-Ahmady El-Naggar

    2014-06-01

    Full Text Available The use of low cost agro-industrial residues for the production of industrial enzymes is one of the ways to reduce significantly production costs. Cellulase producing actinomycetes were isolated from soil and decayed agricultural wastes. Among them, a potential culture, strain NEAE-J, was selected and identified on the basis of morphological, cultural, physiological and chemotaxonomic properties, together with 16S rDNA sequence. It is proposed that strain NEAE-J should be included in the species Streptomyces albogriseolus as a representative of a novel sub-species, Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J and sequencing product was deposited in the GenBank database under accession number JN229412. This organism was tested for its ability to produce endoglucanase and release reducing sugars from agro-industrial residues as substrates. Sugarcane bagasse was the most suitable substrate for endoglucanase production. Effects of process variables, namely incubation time, temperature, initial pH and nitrogen source on production of endoglucanase by submerged fermentation using Streptomyces albogriseolus subsp. cellulolyticus have been studied. Accordingly optimum conditions have been determined. Incubation temperature of 30 ºC after 6 days, pH of 6.5, 1% sugarcane bagasse as carbon source and peptone as nitrogen source were found to be the optimum for endoglucanase production. Optimization of the process parameters resulted in about 2.6 fold increase in the endoglucanase activity. Therefore, Streptomyces albogriseolus subsp. cellulolyticus coud be potential microorganism for the intended application.

  14. Fortunella margarita transcriptional reprogramming triggered by Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Khalaf, Abeer A; Gmitter, Frederick G; Conesa, Ana; Dopazo, Joaquin; Moore, Gloria A

    2011-11-11

    Citrus canker disease caused by the bacterial pathogen Xanthomonas citri subsp. citri (Xcc) has become endemic in areas where high temperature, rain, humidity, and windy conditions provide a favourable environment for the dissemination of the bacterium. Xcc is pathogenic on many commercial citrus varieties but appears to elicit an incompatible reaction on the citrus relative Fortunella margarita Swing (kumquat), in the form of a very distinct delayed necrotic response. We have developed subtractive libraries enriched in sequences expressed in kumquat leaves during both early and late stages of the disease. The isolated differentially expressed transcripts were subsequently sequenced. Our results demonstrate how the use of microarray expression profiling can help assign roles to previously uncharacterized genes and elucidate plant pathogenesis-response related mechanisms. This can be considered to be a case study in a citrus relative where high throughput technologies were utilized to understand defence mechanisms in Fortunella and citrus at the molecular level. cDNAs from sequenced kumquat libraries (ESTs) made from subtracted RNA populations, healthy vs. infected, were used to make this microarray. Of 2054 selected genes on a customized array, 317 were differentially expressed (P < 0.05) in Xcc challenged kumquat plants compared to mock-inoculated ones. This study identified components of the incompatible interaction such as reactive oxygen species (ROS) and programmed cell death (PCD). Common defence mechanisms and a number of resistance genes were also identified. In addition, there were a considerable number of differentially regulated genes that had no homologues in the databases. This could be an indication of either a specialized set of genes employed by kumquat in response to canker disease or new defence mechanisms in citrus. Functional categorization of kumquat Xcc-responsive genes revealed an enhanced defence-related metabolism as well as a number of

  15. Streptococcus equi subsp. zooepidemicus Invades and Survives in Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Bolette Skive

    2017-11-01

    Full Text Available Streptococcus equi subsp. zooepidemicus (S. zooepidemicus is an opportunistic pathogen of several species including humans. S. zooepidemicus is found on mucus membranes of healthy horses, but can cause acute and chronic endometritis. Recently S. zooepidemicus was found able to reside in the endometrium for prolonged periods of time. Thus, we hypothesized that an intracellular phase may be part of the S. zooepidemicus pathogenesis and investigated if S. zooepidemicus was able to invade and survive inside epithelial cells. HEp-2 and HeLa cell lines were co-cultured with two S. zooepidemicus strains (1-4a and S31A1 both originating from the uterus of mares suffering from endometritis. Cells were fixed at different time points during the 23 h infection assay and field emission scanning electron microscopy (FESEM was used to characterize adhesion and invasion mechanisms. The FESEM images showed three morphologically different types of invasion for both bacterial strains. The main port of entry was through large invaginations in the epithelial cell membrane. Pili-like bacterial appendages were observed when the S. zooepidemicus cells were in close proximity to the epithelial cells indicating that attachment and invasion were active processes. Adherent and intracellular S. zooepidemicus, and bacteria in association with lysosomes was determined by immunofluorescence staining techniques and fluorescence microscopy. Quantification of intracellular bacteria was determined in penicillin protection assays. Both S. zooepidemicus strains investigated were able to invade epithelial cells although at different magnitudes. The immunofluorescence data showed significantly higher adhesion and invasion rates for strain 1-4a when compared to strain S31A1. S. zooepidemicus was able to survive intracellularly, but the survival rate decreased over time in the cell culture system. Phagosome-like compartments containing S. zooepidemicus at some stages fused with

  16. Streptococcus equi subsp. zooepidemicus Invades and Survives in Epithelial Cells.

    Science.gov (United States)

    Skive, Bolette; Rohde, Manfred; Molinari, Gabriella; Braunstein, Thomas Hartig; Bojesen, Anders M

    2017-01-01

    Streptococcus equi subsp. zooepidemicus ( S. zooepidemicus ) is an opportunistic pathogen of several species including humans. S. zooepidemicus is found on mucus membranes of healthy horses, but can cause acute and chronic endometritis. Recently S. zooepidemicus was found able to reside in the endometrium for prolonged periods of time. Thus, we hypothesized that an intracellular phase may be part of the S. zooepidemicus pathogenesis and investigated if S. zooepidemicus was able to invade and survive inside epithelial cells. HEp-2 and HeLa cell lines were co-cultured with two S. zooepidemicus strains (1-4a and S31A1) both originating from the uterus of mares suffering from endometritis. Cells were fixed at different time points during the 23 h infection assay and field emission scanning electron microscopy (FESEM) was used to characterize adhesion and invasion mechanisms. The FESEM images showed three morphologically different types of invasion for both bacterial strains. The main port of entry was through large invaginations in the epithelial cell membrane. Pili-like bacterial appendages were observed when the S. zooepidemicus cells were in close proximity to the epithelial cells indicating that attachment and invasion were active processes. Adherent and intracellular S. zooepidemicus , and bacteria in association with lysosomes was determined by immunofluorescence staining techniques and fluorescence microscopy. Quantification of intracellular bacteria was determined in penicillin protection assays. Both S. zooepidemicus strains investigated were able to invade epithelial cells although at different magnitudes. The immunofluorescence data showed significantly higher adhesion and invasion rates for strain 1-4a when compared to strain S31A1. S. zooepidemicus was able to survive intracellularly, but the survival rate decreased over time in the cell culture system. Phagosome-like compartments containing S. zooepidemicus at some stages fused with lysosomes to form a

  17. Bioaccessible Antioxidants in Milk Fermented by Bifidobacterium longum subsp. longum Strains

    Science.gov (United States)

    Gagnon, Mérilie; Savard, Patricia; Rivière, Audrey; LaPointe, Gisèle

    2015-01-01

    Bifidobacterium longum subsp. longum is among the dominant species of the human gastrointestinal microbiota and could thus have potential as probiotics. New targets such as antioxidant properties have interest for beneficial effects on health. The objective of this study was to evaluate the bioaccessibility of antioxidants in milk fermented by selected B. longum subsp. longum strains during in vitro dynamic digestion. The antioxidant capacity of cell extracts from 38 strains, of which 32 belong to B. longum subsp. longum, was evaluated with the ORAC (oxygen radical absorbance capacity) method. On the basis of screening and gene sequence typing by multilocus locus sequence analysis (MLSA), five strains were chosen for fermenting reconstituted skim milk. Antioxidant capacity varied among the strains tested (P = 0.0009). Two strains of B. longum subsp. longum (CUETM 172 and 171) showed significantly higher ORAC values than the other bifidobacteria strains. However, there does not appear to be a relationship between gene sequence types and antioxidant capacity. The milk fermented by each of the five strains selected (CUETM 268, 172, 245, 247, or PRO 16-10) did not have higher initial ORAC values compared to the nonfermented milk samples. However, higher bioaccessibility of antioxidants in fermented milk (175–358%) was observed during digestion. PMID:25802836

  18. Bioaccessible antioxidants in milk fermented by Bifidobacterium longum subsp. longum strains.

    Science.gov (United States)

    Gagnon, Mérilie; Savard, Patricia; Rivière, Audrey; LaPointe, Gisèle; Roy, Denis

    2015-01-01

    Bifidobacterium longum subsp. longum is among the dominant species of the human gastrointestinal microbiota and could thus have potential as probiotics. New targets such as antioxidant properties have interest for beneficial effects on health. The objective of this study was to evaluate the bioaccessibility of antioxidants in milk fermented by selected B. longum subsp. longum strains during in vitro dynamic digestion. The antioxidant capacity of cell extracts from 38 strains, of which 32 belong to B. longum subsp. longum, was evaluated with the ORAC (oxygen radical absorbance capacity) method. On the basis of screening and gene sequence typing by multilocus locus sequence analysis (MLSA), five strains were chosen for fermenting reconstituted skim milk. Antioxidant capacity varied among the strains tested (P = 0.0009). Two strains of B. longum subsp. longum (CUETM 172 and 171) showed significantly higher ORAC values than the other bifidobacteria strains. However, there does not appear to be a relationship between gene sequence types and antioxidant capacity. The milk fermented by each of the five strains selected (CUETM 268, 172, 245, 247, or PRO 16-10) did not have higher initial ORAC values compared to the nonfermented milk samples. However, higher bioaccessibility of antioxidants in fermented milk (175-358%) was observed during digestion.

  19. Multilocus sequence typing reveals two evolutionary lineages of Acidovorax avenae subsp. citrulli.

    Science.gov (United States)

    Feng, Jianjun; Schuenzel, Erin L; Li, Jianqiang; Schaad, Norman W

    2009-08-01

    Acidovorax avenae subsp. citrulli, causal agent of bacterial fruit blotch, has caused considerable damage to the watermelon and melon industry in China and the United States. Understanding the emergence and spread of this pathogen is important for controlling the disease. To build a fingerprinting database for reliable identification and tracking of strains of A. avenae subsp. citrulli, a multilocus sequence typing (MLST) scheme was developed using seven conserved loci. The study included 8 original strains from the 1978 description of A. avenae subsp. citrulli, 51 from China, and 34 from worldwide collections. Two major clonal complexes (CCs), CC1 and CC2, were identified within A. avenae subsp. citrulli; 48 strains typed as CC1 and 45 as CC2. All eight original 1978 strains isolated from watermelon and melon grouped in CC1. CC2 strains were predominant in the worldwide collection and all but five were isolated from watermelon. In China, a major seed producer for melon and watermelon, the predominant strains were CC1 and were found nearly equally on melon and watermelon.

  20. Genome sequence of the rice-pathogenic bacterium Acidovorax avenae subsp. avenae RS-1.

    Science.gov (United States)

    Xie, Guan-Lin; Zhang, Guo-Qing; Liu, He; Lou, Miao-Miao; Tian, Wen-Xiao; Li, Bin; Zhou, Xue-Ping; Zhu, Bo; Jin, Gu-Lei

    2011-09-01

    Acidovorax avenae subsp. avenae is a phytobacterium which is the causative agent of several plant diseases with economic significance. Here, we present the draft genome sequence of strain RS-1, which was isolated from rice shoots in a rice field in China. This strain can cause bacterial stripe of rice. Copyright © 2011, American Society for Microbiology. All Rights Reserved.

  1. Genome Sequence of the Rice-Pathogenic Bacterium Acidovorax avenae subsp. avenae RS-1 ▿

    OpenAIRE

    Xie, Guan-Lin; Zhang, Guo-Qing; Liu, He; Lou, Miao-Miao; Tian, Wen-Xiao; Li, Bin; Zhou, Xue-Ping; Zhu, Bo; Jin, Gu-Lei

    2011-01-01

    Acidovorax avenae subsp. avenae is a phytobacterium which is the causative agent of several plant diseases with economic significance. Here, we present the draft genome sequence of strain RS-1, which was isolated from rice shoots in a rice field in China. This strain can cause bacterial stripe of rice.

  2. Genome Sequence of the Rice-Pathogenic Bacterium Acidovorax avenae subsp. avenae RS-1 ▿

    Science.gov (United States)

    Xie, Guan-Lin; Zhang, Guo-Qing; Liu, He; Lou, Miao-Miao; Tian, Wen-Xiao; Li, Bin; Zhou, Xue-Ping; Zhu, Bo; Jin, Gu-Lei

    2011-01-01

    Acidovorax avenae subsp. avenae is a phytobacterium which is the causative agent of several plant diseases with economic significance. Here, we present the draft genome sequence of strain RS-1, which was isolated from rice shoots in a rice field in China. This strain can cause bacterial stripe of rice. PMID:21742879

  3. Biological Control to Protect Watermelon Blossoms and Seed from Infection by Acidovorax avenae subsp. citrulli.

    Science.gov (United States)

    Fessehaie, A; Walcott, R R

    2005-04-01

    ABSTRACT The efficacy of biological control seed treatments with Pseudomonas fluorescens (A506), Acidovorax avenae subsp. avenae (AAA 99-2), and an unidentified gram-positive bacterium recovered from watermelon seed (WS-1) was evaluated for the management of bacterial fruit blotch (BFB) of watermelon. In growth chamber and greenhouse experiments, seed treated with AAA 99-2 displayed superior disease suppression, reducing BFB transmission by 96.5%. AAA 99-2, P. fluorescens A506, and Kocide also suppressed the epiphytic growth of A. avenae subsp. citrulli when applied to attached watermelon blossoms 5 h prior to inoculation. Watermelon blossom protection reduced seed infestation by A. avenae subsp. citrulli. From blossoms treated with 0.1 M phosphate buffered saline (PBS), 63% of the resulting seed lots were infested with A. avenae subsp. citrulli. In contrast, for blossoms protected with WS-1, Kocide, P. fluorescens A506, and AAA 99-2, the proportion of infested seed lots were 48.3, 21.1, 24.1, and 13.8%, respectively. The effect of blossom treatments on seed lot infestation was statistically significant (P = 0.001) but WS-1 was not significantly different from PBS. These findings suggest that blossom protection with biological control agents could be a feasible option for managing BFB.

  4. Demodicosis in Chamois ( Rupicapra rupicapra subsp. rupicapra) in the Italian Alps, 2013-14.

    Science.gov (United States)

    Salvadori, Claudia; Formenti, Nicoletta; Trogu, Tiziana; Lanfranchi, Paolo; Papini, Roberto A; Poli, Alessandro

    2016-04-28

    We report demodicosis in five alpine chamois ( Rupicapra rupicapra subsp. rupicapra) from the Italian Alps that showed moderate crusts on the head and dorsal aspect of the trunk. We detected intramural folliculitis, moderate dermatitis, and T-lymphocytes and macrophages associated with Demodex spp. in follicles and sebaceous glands.

  5. Complete genome sequence of the prototype lactic acid bacterium Lactococcus lactis subsp cremoris MG1363

    NARCIS (Netherlands)

    Wegmann, Udo; O'Connell-Motherwy, Mary; Zomer, Aldert; Buist, Girbe; Shearman, Claire; Canchaya, Carlos; Ventura, Marco; Goesmann, Alexander; Gasson, Michael J.; Kuipers, Oscar P.; van Sinderen, Douwe; Kok, Jan

    Lactococcus lactis is of great importance for the nutrition of hundreds of millions of people worldwide. This paper describes the genome sequence of Lactococcus lactis subsp. cremoris MG1363, the lactococcal strain most intensively studied throughout the world. The 2,529,478-bp genome contains 81

  6. Introduction of peptidase genes from Lactobacillus delbrueckii subsp. lactis into Lactococcus lactis and controlled expression

    NARCIS (Netherlands)

    Wegmann, U.; Klein, J.R.; Drumm, I.; Kuipers, O.P.; Henrich, B.

    1999-01-01

    Peptidases PepI, PepL, PepW, and PepG from Lactobacillus delbrueckii subsp, lactis, which have no counterparts in Lactococcus lactis, and peptidase PepQ were examined to determine their potential to confer new peptidolytic properties to lactococci, Controllable expression of the corresponding genes

  7. Resistance of sweet orange Pera (Citrus sinensis) genotypes to Xanthomonas citri subsp. citri under field conditions

    Science.gov (United States)

    Citrus canker control is based on protection measures and eradication of plants infected with Xanthomonas citri subsp. citri. Although these measures show satisfactory results, the use of resistant genotypes is an important alternative for citrus canker control. The aim of this study was to evaluate...

  8. Inferring biomarkers for Mycobacterium avium subsp. paratuberculosis infection and disease progression using experimental data

    Science.gov (United States)

    Available diagnostic assays for Mycobacterium avium subsp paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of the infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN- gamma, ELI...

  9. Vaccination with peptides of Mycobacterium avium subsp. paratuberculosis (MAP) reduces MAP burden of infected goats

    DEFF Research Database (Denmark)

    Melvang, Heidi Mikkelsen; Hassan, Sufia Butt; Thakur, Aneesh

    Mycobacterium avium subsp. paratuberculosis (Map) is the cause of paratuberculosis, a chronic enteritis of ruminants that is widespread worldwide. We investigated the effect of post-exposure vaccination with Map specific peptides in a goat model aiming at developing a Map vaccine that will neither...

  10. Detection of Mycobacterium avium subsp. paratuberculosis in Drinking Water and Biofilms Using Quantitative PCR

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s disease in domestic animals and has been implicated in Crohn’s disease in humans. Cows infected with Johne’s disease shed large quantities of MAP into soil. Further, MAP has been isolated from surface water, is resi...

  11. Detection of Mycobacterium avium subsp. paratuberculosis in milk from clinically affected cows by PCR and culture

    DEFF Research Database (Denmark)

    Giese, Steen Bjørck; Ahrens, Peter

    2000-01-01

    Milk and faeces samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by culture and PCR. M. paratuberculosis was cultivated in variable numbers from faeces or intestinal mucosa in eight of 11...

  12. Transcriptome profiling of Lactococcus lactis subsp. cremoris CECT 8666 in response to agmatine

    NARCIS (Netherlands)

    Del Rio, Beatriz; Redruello, Begoña; Martin, M Cruz; Fernandez, Maria; de Jong, Anne; Kuipers, Oscar P; Ladero, Victor; Alvarez, Miguel A

    2016-01-01

    The dairy strain Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14) synthesizes the biogenic amine putrescine from agmatine via the agmatine deiminase (AGDI) pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are

  13. Complete genomic sequence of campylobacter jejuni subsp. jejuni HS:19 penner reference strain

    Science.gov (United States)

    Campylobacter jejuni subsp. jejuni (Cjj) infections are a leading cause of foodborne gastroenteritis and the most prevalent antecedent to Guillain-Barré syndrome (GBS). Capsular type Penner HS:19 is among several capsule types shown to be markers for GBS. This study describes the genome of Cjj HS:19...

  14. An original case of Francisella tularensis subsp. holarctica bacteremia after a near-drowning accident.

    Science.gov (United States)

    Ughetto, Estelle; Héry-Arnaud, Geneviève; Cariou, Marie-Estelle; Pelloux, Isabelle; Maurin, Max; Caillon, Jocelyne; Moreau, Philippe; Ygout, Jean-François; Corvec, Stéphane

    2015-08-01

    We report the first case of Francisella tularensis subsp. holarctica bacteremia after water contamination in France. A 75-year-old man developed septic pneumonic tularemia after a near-drowning accident. We highlight the need for a longer incubation time for isolation of F. tularensis from blood cultures.

  15. Draft Genome Sequence of Lactobacillus delbrueckii subsp. bulgaricus LBB.B5

    NARCIS (Netherlands)

    Urshev, Z.; Hajo, K.; Lenoci, L.; Bron, P.A.; Dijkstra, A.; Alkema, W.; Wels, M.; Siezen, R.J.; Minkova, S.; Hijum, S.A. van

    2016-01-01

    Lactobacillus delbrueckii subsp. bulgaricus LBB.B5 originates from homemade Bulgarian yogurt and was selected for its ability to form a strong association with Streptococcus thermophilus The genome sequence will facilitate elucidating the genetic background behind the contribution of LBB.B5 to the

  16. Factors Affecting Exocellular Polysaccharide Production by Lactobacillus delbrueckii subsp. bulgaricus Grown in a Chemically Defined Medium†

    Science.gov (United States)

    Petry, Sandrine; Furlan, Sylviane; Crepeau, Marie-Jeanne; Cerning, Jutta; Desmazeaud, Michel

    2000-01-01

    We developed a chemically defined medium (CDM) containing lactose or glucose as the carbon source that supports growth and exopolysaccharide (EPS) production of two strains of Lactobacillus delbrueckii subsp. bulgaricus. The factors found to affect EPS production in this medium were oxygen, pH, temperature, and medium constituents, such as orotic acid and the carbon source. EPS production was greatest during the stationary phase. Composition analysis of EPS isolated at different growth phases and produced under different fermentation conditions (varying carbon source or pH) revealed that the component sugars were the same. The EPS from strain L. delbrueckii subsp. bulgaricus CNRZ 1187 contained galactose and glucose, and that of strain L. delbrueckii subsp. bulgaricus CNRZ 416 contained galactose, glucose, and rhamnose. However, the relative proportions of the individual monosaccharides differed, suggesting that repeating unit structures can vary according to specific medium alterations. Under pH-controlled fermentation conditions, L. delbrueckii subsp. bulgaricus strains produced as much EPS in the CDM as in milk. Furthermore, the relative proportions of individual monosaccharides of EPS produced in pH-controlled CDM or in milk were very similar. The CDM we developed may be a useful model and an alternative to milk in studies of EPS production. PMID:10919802

  17. Lymphoproliferative and gamma interferon responses to stress-regulated Mycobacterium avium subsp. paratuberculosis recombinant proteins

    Science.gov (United States)

    Johne’s disease in ruminants is a chronic infection of the intestines caused by Mycobacterium avium subsp. paratuberculosis. Economic losses associated with Johne’s disease arise due to premature culling, reduced production of milk and wool and mortalities. The disease is characterised by a long inc...

  18. Mycobacterium avium Subsp. avium Infection in Four Veal Calves: Differentiation from Intestinal Tuberculosis

    Directory of Open Access Journals (Sweden)

    Christine Goepfert

    2014-01-01

    Full Text Available Mycobacterium avium subsp. avium (Maa is an intracellular pathogen belonging to the Mycobacterium avium-intracellulare complex (MAC. Reservoirs of MAC are the natural environment, wildlife and domestic animals. In adult bovine, MAC infections are typically caused by Mycobacterium avium subsp. paratuberculosis (Map. Maa infections in bovine are rarely reported but may cause clinical disease and pathological lesions similar to those observed in paratuberculosis or those induced by members of the Mycobacterium tuberculosis complex (MTBC. Therefore, differentiation of MAC from MTBC infection should be attempted, especially if unusual mycobacterial lesions are encountered. Four veal calves from a fattening farm dying with clinical signs of otitis media, fever, and weight loss were submitted for necropsy. Samples from affected organs were taken for histologic investigation, bacteriologic culture, and bacterial specification using PCR. Macroscopic thickening of the intestinal mucosa was induced by granulomatous enteritis and colitis. Intracytoplasmic acid-fast bacteria were detected by Ziehl-Neelsen stains and PCR revealed positive results for Mycobacterium avium subsp. avium. Clinical and pathological changes of Maa infection in veal calves had features of Mycobacterium avium subsp. paratuberculosis and the MTBC. Therefore, Mycobacterium tuberculosis complex infection should be considered in cases of granulomatous enteritis in calves.

  19. Optimization of hexadecylpyridinium chloride decontamination for culture of Mycobacterium avium subsp. paratuberculosis from milk

    Science.gov (United States)

    Cows in advanced stages of Johne’s disease shed Mycobacterium avium subsp. paratuberculosis (MAP) into both their milk and feces, allowing for transmission of the bacteria between animals. The objective of this study was to formulate an optimized protocol for the isolation of MAP from milk and colos...

  20. Different Mycobacterium avium subsp. paratuberculosis MIRU-VNTR patterns coexist within cattle herds

    NARCIS (Netherlands)

    Hulzen, van K.J.E.; Heuven, H.C.M.; Nielen, M.; Hoeboer, J.; Santema, W.J.; Koets, A.P.

    2011-01-01

    A better understanding of the biodiversity of Mycobacterium avium subsp. paratuberculosis (MAP) offers more insight in the epidemiology of paratuberculosis and therefore may contribute to the control of the disease. The aim of this study was to investigate the genetic diversity in bovine MAP

  1. Characterization of cry1Cb3 and cry1Fb7 from Bacillus thuringiensis subsp. galleriae

    Czech Academy of Sciences Publication Activity Database

    Huang, T.; Xiao, Y.; Pan, J.; Zhang, L.; Gelbič, Ivan; Guan, X.

    2015-01-01

    Roč. 10, č. 1 (2015), s. 521-528 ISSN 2391-5412 Institutional support: RVO:60077344 Keywords : Bacillus thuringiensis subsp. galleriae * PCR-RFLP * cloning Subject RIV: EB - Genetics ; Molecular Biology http://www.degruyter.com/view/j/biol.2015.10.issue-1/biol-2015-0054/biol-2015-0054.xml

  2. Genome Sequence of Lactococcus lactis subsp. lactis NCDO 2118, a GABA-Producing Strain

    DEFF Research Database (Denmark)

    Oliveira, Letícia C; Saraiva, Tessália D L; Soares, Siomar C

    2014-01-01

    Lactococcus lactis subsp. lactis NCDO 2118 is a nondairy lactic acid bacterium, a xylose fermenter, and a gamma-aminobutyric acid (GABA) producer isolated from frozen peas. Here, we report the complete genome sequence of L. lactis NCDO 2118, a strain with probiotic potential activity....

  3. Environmental contamination with Mycobacterium avium subsp. paratuberculosis in endemically infected dairy herds

    Science.gov (United States)

    Environmental contamination with Mycobacterium avium subsp. paratuberculosis (MAP) is thought to be the primary source of infection for dairy cattle. The exact link between fecal shedding of MAP by individual cows and environmental contamination levels at the herd level was explored with a cross-se...

  4. Tulum Peynirlerinden izole Edilen Lactococcus lactis subsp. lactis YBML9 ve

    Directory of Open Access Journals (Sweden)

    Yasin TUNCER

    2009-04-01

    Full Text Available Bu çalısmanın amacı tulum peynirlerinden izole edilen Lactococcus lactis suslarının fenotipik tanısı ve bu suslar tarafından üretilen bakteriyosinlerin kısmi karakterizasyonlarıdır. Bu amaçla Türkiye'nin sekiz farklı ilinden (Ankara, Antalya, Burdur, Denizli, Erzincan, Isparta, İstanbul ve İzmir yöresel pazarlardan toplanan 60 adet tulum peyniri örneginden 40 adet Lactococcus lactis susu (31 adet L. lactis subsp. lactis ve 9 adet L. lactis subsp. cremoris izole edildi. 40 adet L. lactis susu içerisinden, 2 adet L. lactis subsp. lactis (YBML9 ve YBML21 susu bakteriyosin üretme yeteneginde bulundu. L. lactis subsp. lactis YBML9 ve YBML21 susları tarafından üretilen bakteriyosinler, farklı enzim, pH ve sıcaklık uygulamaları sonucu; sırasıyla nisin ve laktisin 481 olarak tanımlandı.

  5. Prophage lysin Ply30 protects mice from Streptococcus suis and Streptococcus equi subsp. zooepidemicus infections.

    Science.gov (United States)

    Tang, Fang; Li, Dezhi; Wang, Haojin; Ma, Zhe; Lu, Chengping; Dai, Jianjun

    2015-11-01

    Streptococcus suis and Streptococcus equi subsp. zooepidemicus are capable of infecting humans and various animals, causing significant problems for the worldwide swine industry. As antibiotic resistance has increased, lysosomal enzymes encoded by phages have shown potential for use against pathogenic bacteria. In this study, a novel bacteriophage lysin, Ply30, encoded by the S. suis prophage phi30c, was recombinantly expressed and purified. Ply30 showed high bacteriolysis activity on S. suis and S. equi subsp. zooepidemicus in vitro. The ratio of the optical density at 600 nm (OD600) with treatment versus the OD600 with no treatment for most tested S. suis and S. equi subsp. zooepidemicus strains decreased from 1 to Streptococcus strains ranged from 16 to 512 μg/ml. In vivo, a 2-mg dose of Ply30 protected 90% (9/10 mice) of mice from infection with S. equi subsp. zooepidemicus and 80% (8/10 mice) of mice from infection with S. suis. Seven days after lysin Ply30 treatment, bacterial loads were significantly decreased in all tested organs and blood compared with those at 1 h postinfection without Ply30 treatment. Ply30 showed in vitro and in vivo antimicrobial efficiency and protected mice against two kinds of bacterial infections, indicating that Ply30 may be an effective therapeutic against streptococci. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  6. First identification of Francisella noatunensis subsp. orientalis causing mortality in Mexican tilapia Oreochromis spp.

    Science.gov (United States)

    Ortega, Cesar; Mancera, Gerardo; Enríquez, Ricardo; Vargas, Augusto; Martínez, Simón; Fajardo, Raúl; Avendaño-Herrera, Ruben; Navarrete, María José; Romero, Alex

    2016-08-09

    Francisellosis, an emerging disease in tilapia Oreochromis spp., is caused by the facultative, intracellular bacterium Francisella noatunensis subsp. orientalis, which is present in various countries where tilapia farming is commercially important. We confirmed the presence of francisellosis in Mexican tilapia cultures in association with an outbreak during the second semester of 2012. Broodstock fish presented a mortality rate of approximately 40%, and disease was characterized by histologically classified granulomas, or whitish nodules, in different organs, mainly the spleen and kidney. Through DNA obtained from infected tissue and pure cultures in a cysteine heart medium supplemented with hemoglobin, F. noatunensis subsp. orientalis was initially confirmed through the amplification and analysis of the 16S rRNA gene and the internal transcribed spacer region. Phylogenetic analysis of these genes demonstrated close similarity with previously reported F. noatunensis subsp. orientalis sequences obtained from infected tilapia from various countries. The identification of this subspecies as the causative agent of the outbreak was confirmed using the iglC gene as a target sequence, which showed 99.5% identity to 2 F. noatunensis subsp. orientalis strains (Ethime-1 and Toba04). These findings represent the first documented occurrence of francisellosis in Mexican tilapia cultures, which highlights the importance of establishing preventative measures to minimize the spread of this disease within the Mexican aquaculture industry.

  7. PURIFICATION AND CHARACTERIZATION OF AN ENDOPEPTIDASE FROM LACTOCOCCUS-LACTIS SUBSP CREMORIS WG2

    NARCIS (Netherlands)

    TAN, PST; POS, KM; KONINGS, WN

    1991-01-01

    An endopeptidase has been purified to homogeneity from a crude cell extract of Lactococcus lactis subsp. cremoris Wg2 by a procedure that includes diethyl-aminoethane-Sephacel chromatography, phenyl-Sepharose chromatography, hydroxylapatite chromatography, and fast protein liquid chromatography over

  8. Geography of genetic differentiation in the barley wild relative Hordeum vulgare subsp. spontaneum in Jordan

    Science.gov (United States)

    Informed collecting, conservation, monitoring and utilization of genetic diversity require knowledge of the distribution and structure of genetic variation occurring in a species. Hordeum vulgare subsp. spontaneum (K. Koch) Thell., a primary wild relative of barley, is an important source of genetic...

  9. Phylogenomic analysis shows that Bacillus amyloliquefaciens subsp. plantarum is a later heterotypic synonym of Bacillus methylotrophicus.

    Science.gov (United States)

    Dunlap, Christopher A; Kim, Soo-Jin; Kwon, Soon-Wo; Rooney, Alejandro P

    2015-07-01

    The rhizosphere-isolated bacteria belonging to the Bacillus amyloliquefaciens subsp. plantarum and Bacillus methylotrophicus clades are an important group of strains that are used as plant growth promoters and antagonists of plant pathogens. These properties have made these strains the focus of commercial interest. Here, we present the draft genome sequence of B. methylotrophicus KACC 13105(T) ( = CBMB205(T)). Comparative genomic analysis showed only minor differences between this strain and the genome of the B. amyloliquefaciens subsp. plantarum type strain, with the genomes sharing approximately 95% of the same genes. The results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the type strains of these two taxa are highly similar. In fact, our results show that the type strain of B. amyloliquefaciens subsp. plantarum FZB42(T) ( = DSM 23117(T) = BGSC 10A6(T)) does not cluster with other members of the B. amyloliquefaciens taxon. Instead, it clusters well within a clade of strains that are assigned to B. methylotrophicus, including the type strain of that species. Therefore, we propose that the subspecies B. amyloliquefaciens subsp. plantarum should be reclassified as a later heterotypic synonym of B. methylotrophicus.

  10. Pectobacterium carotovorum subsp carotovorum can cause potato blackleg in temperate climates

    NARCIS (Netherlands)

    Haan, de E.G.; Dekker-Nooren, T.C.E.M.; Bovenkamp, van den G.W.; Speksnijder, A.G.C.L.; Zouwen, van der P.S.; Wolf, van der J.M.

    2008-01-01

    It is well established that the pectinolytic bacteria Pectobacterium atrosepticum (Pca) and Dickeya spp. are causal organisms of blackleg in potato. In temperate climates, the role of Pectobacterium carotovorum subsp. carotovorum (Pcc) in potato blackleg, however, is unclear. In different western

  11. Distribution of Dickeya spp. and Pectobacterium carotovorum subsp carotovorum in naturally infected seed potatoes

    NARCIS (Netherlands)

    Czajkowski, R.L.; Grabe, G.J.; Wolf, van der J.M.

    2009-01-01

    Detailed studies were conducted on the distribution of Pectobacterium carotovorum subsp. carotovorum and Dickeya spp. in two potato seed lots of different cultivars harvested from blackleg-diseased crops. Composite samples of six different tuber sections (peel, stolon end, and peeled potato tissue

  12. Complete Genome Sequence of Mycobacterium fortuitum subsp. fortuitum Type Strain DSM46621

    KAUST Repository

    Ho, Y. S

    2012-10-26

    Mycobacterium fortuitum is a member of the rapidly growing nontuberculous mycobacteria (NTM). It is ubiquitous in water and soil habitats, including hospital environments. M. fortuitum is increasingly recognized as an opportunistic nosocomial pathogen causing disseminated infection. Here we report the genome sequence of M. fortuitum subsp. fortuitum type strain DSM46621.

  13. Prophage Lysin Ply30 Protects Mice from Streptococcus suis and Streptococcus equi subsp. zooepidemicus Infections

    Science.gov (United States)

    Tang, Fang; Li, Dezhi; Wang, Haojin; Ma, Zhe; Lu, Chengping

    2015-01-01

    Streptococcus suis and Streptococcus equi subsp. zooepidemicus are capable of infecting humans and various animals, causing significant problems for the worldwide swine industry. As antibiotic resistance has increased, lysosomal enzymes encoded by phages have shown potential for use against pathogenic bacteria. In this study, a novel bacteriophage lysin, Ply30, encoded by the S. suis prophage phi30c, was recombinantly expressed and purified. Ply30 showed high bacteriolysis activity on S. suis and S. equi subsp. zooepidemicus in vitro. The ratio of the optical density at 600 nm (OD600) with treatment versus the OD600 with no treatment for most tested S. suis and S. equi subsp. zooepidemicus strains decreased from 1 to Streptococcus strains ranged from 16 to 512 μg/ml. In vivo, a 2-mg dose of Ply30 protected 90% (9/10 mice) of mice from infection with S. equi subsp. zooepidemicus and 80% (8/10 mice) of mice from infection with S. suis. Seven days after lysin Ply30 treatment, bacterial loads were significantly decreased in all tested organs and blood compared with those at 1 h postinfection without Ply30 treatment. Ply30 showed in vitro and in vivo antimicrobial efficiency and protected mice against two kinds of bacterial infections, indicating that Ply30 may be an effective therapeutic against streptococci. PMID:26253669

  14. The anti-tick properties of the root extracts of Senna italica subsp ...

    African Journals Online (AJOL)

    SERVER

    2007-11-15

    Nov 15, 2007 ... This study examined the anti-tick properties of the root extracts of Senna italica subsp. arachoides against adults of Hyalomma marginatum rufipes. Of the hexane, chloroform, dichloromethane, ethyl acetate and methanol extracts tested, only ethyl acetate extracts proved to be potent against adults of.

  15. Draft Genome Sequence of Xylella fastidiosa subsp. fastidiosa Strain Stag?s Leap

    OpenAIRE

    Chen, J.; Wu, F.; Zheng, Z.; Deng, X.; Burbank, L. P.; Stenger, D. C.

    2016-01-01

    Xylella fastidiosa subsp. fastidiosa causes Pierce?s disease of grapevine. Presented here is the draft genome sequence of the Stag?s Leap strain, previously used in pathogenicity/virulence assays to evaluate grapevine germplasm bearing Pierce?s disease resistance and a phenotypic assessment of knockout mutants to determine gene function.

  16. Draft genome sequence of Xylella fastidiosa subsp. fastidiosa strain Stag’s Leap

    Science.gov (United States)

    Xylella fastidiosa subsp. fastidiosa causes Pierce’s disease of grapevine. Presented here is the draft genome sequence of the Stag’s Leap strain, previously used in pathogenicity/virulence assays to evaluate grapevine germplasm bearing Pierce’s disease....

  17. Studies upon morhological and biological traits of Festuca rubra, subsp.fallax (Poaceae

    Directory of Open Access Journals (Sweden)

    Bogusław Sawicki

    2013-12-01

    Full Text Available Observation and measurements of some traits of Festuca rubra L., subsp. fallax (Thuill. Hack. ecotypes were made in 1995-1997 using samples selected from natural habitats and collected in Grassland Experimental Station in Sosnowica. High differentiation of traits under study and their correlations were found. Valorized ecotypes are good material for new varieties breeding.

  18. Draft Genome Sequences of 64 Salmonella enterica subsp. enterica Enteritidis Isolates from Mice in US

    Science.gov (United States)

    A ciprofloxacin resistant (CipR) Salmonella enterica subsp. enterica serovar Kentucky ST198 has rapidly and extensively disseminated globally to become a major food-safety and public health concern. Here, we report a complete genome sequence of a CipR S. Kentucky ST198 strain PU131 isolated from a ...

  19. Selective enumeration and viability of Bifidobacterium animalis subsp. lactis in a new fermented milk product Enumeração seletiva e viabilidade de Bifidobacterium animalis subsp. lactis em um novo produto lácteo fermentado

    Directory of Open Access Journals (Sweden)

    Adriane Elisabete Costa Antunes

    2007-03-01

    Full Text Available One of the key focuses of today's dairy industry worldwide is the continued development of new products, especially probiotic-based products. Buttermilk is originally a by-product of butter making fermented by Mesophilic Aromatic Cultures (MAC. It can also be made by fermentation of pasteurized whole milk or skimmed milk. This product is not marketed in Brazil. The objectives of this work were: (1 to develop a selective medium for Bifidobacterium animalis subsp. lactis enumeration and (2 to determine the viability of this microorganism during the shelf life of the buttermilk. Skim milk added with 10% sucrose or 0.03% sucralose was pasteurized and inoculated with a composite starter culture consisting of 1% MAC (containing Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis biovar. diacetylactis and Leuconostoc mesenteroides subsp. cremoris and 2% Bifidobacterium animalis subsp. lactis. To attain selective counts of Bif. animalis subsp. lactis the MRS agar supplemented with 0.5% L-cysteine hydrochloride at 10%, 1% lithium chloride at 10%, 0.01% aniline blue and 0.5% dicloxacillin at 0.1% was modified by increasing the antibiotic concentration, addition of NaCl, adjusting pH to 4.8 or increasing the incubation temperature (from 37 to 45ºC. Raising the incubation temperature to 45ºC was found to be efficient in inhibiting the MAC cultures, even in media not added with dicloxacillin. Bif. animalis subsp. lactis exhibited high viability in the product. The buttermilk product prepared with sucrose and sweetener contained in excess of 10(8 cfu.ml-1 bifidobacteria throughout the shelf life of the product (28 days.Atualmente, um dos principais focos da indústria de laticínios em todo o mundo é o desenvolvimento de novos produtos, especialmente probióticos. Buttermilk é originalmente um sub-produto do processamento da manteiga fermentado por Culturas Aromáticas Mesofílicas (MAC. Pode também ser

  20. Is the Evolution of Salmonella enterica subsp. enterica Linked to Restriction-Modification Systems?

    Science.gov (United States)

    Hendriksen, Rene S.; Leekitcharoenphon, Pimlapas; Lukjancenko, Oksana; Hasman, Henrik; Aarestrup, Frank M.

    2016-01-01

    ABSTRACT Salmonella enterica subsp. enterica bacteria are highly diverse foodborne pathogens that are subdivided into more than 1,500 serovars. The diversity is believed to result from mutational evolution, as well as intra- and interspecies recombination that potentially could be influenced by restriction-modification (RM) systems. The aim of this study was to investigate whether RM systems were linked to the evolution of Salmonella enterica subsp. enterica. The study included 221 Salmonella enterica genomes, of which 68 were de novo sequenced and 153 were public available genomes from ENA. The data set covered 97 different serovars of Salmonella enterica subsp. enterica and an additional five genomes from four other Salmonella subspecies as an outgroup for constructing the phylogenetic trees. The phylogenetic trees were constructed based on multiple alignment of core genes, as well as the presence or absence of pangenes. The topology of the trees was compared to the presence of RM systems, antimicrobial resistance (AMR) genes, Salmonella pathogenicity islands (SPIs), and plasmid replicons. We did not observe any correlation between evolution and the RM systems in S. enterica subsp. enterica. However, sublineage correlations and serovar-specific patterns were observed. Additionally, we conclude that plasmid replicons, SPIs, and AMR were all better correlated to serovars than to RM systems. This study suggests a limited influence of RM systems on the evolution of Salmonella enterica subsp. enterica, which could be due to the conjugational mode of horizontal gene transfer in Salmonella. Thus, we conclude that other factors must be involved in shaping the evolution of bacteria. IMPORTANCE The evolution of bacterial pathogens, their plasticity and ability to rapidly change and adapt to new surroundings are crucial for understanding the epidemiology and public health. With the application of genomics, it became clear that horizontal gene transfer played a key role in

  1. Pantoea stewartii subsp. stewartii: lessons learned from a xylem-dwelling pathogen of sweet corn.

    Science.gov (United States)

    Roper, M Caroline

    2011-09-01

    Pantoea stewartii subsp. stewartii is a Gram-negative enteric bacterium that primarily infects sweet corn. Studies of this bacterium have provided useful insight into how xylem-dwelling bacteria establish themselves and incite disease in their hosts. Pantoea stewartii subsp. stewartii is a remarkable bacterial system for laboratory studies because of its relative ease of propagation and genetic manipulation, and the fact that it appears to employ a minimal number of pathogenicity mechanisms. In addition, P. stewartii subsp. stewartii produces copious amounts of its quorum sensing (QS) signal, acyl-homoserine lactone (AHL), making it an excellent organism for studying QS-controlled gene regulation in a plant-pathogenic bacterium. In fact, P. stewartii subsp. stewartii has become the microbial paradigm for QS control of gene expression by both repression and activation via a QS regulator that binds DNA in the absence and dissociates in the presence of the signal ligand. Moreover, P. stewartii subsp. stewartii is a member of the Enterobacteriaceae, and lessons learned from its interaction with plants may be extrapolated to other plant-associated enterics, such as Erwinia, Dickeya and Pectobacterium spp., or enteric human pathogens associated with plants, such as Escherichia coli and Salmonella spp. Bacteria; Gammaproteobacteria; family Enterobacteriaceae; genus Pantoea; species stewartii (Mergaert et al., 1993). Gram-negative, motile, yellow pigmented, mucoid, facultative anaerobe. Pantoea stewartii subsp. stewartii (Smith, 1898) Dye causes Stewart's wilt of corn (Zea mays). Early-maturing sweet corn varieties and some elite inbred maize lines are particularly susceptible. There are two major phases of Stewart's wilt disease: (i) wilt and (ii) leaf blight. The wilt phase occurs when young seedlings are infected with P. stewartii subsp. stewartii (Fig. 1A). Water-soaked lesions first appear on the young expanding leaves and, later, seedlings may become severely wilted

  2. Effect of Salicylic Acid on the Growth and Chemical Responses of Pectobacterium carotovorum subsp. carotovorum.

    Science.gov (United States)

    Ntushelo, Khayalethu

    2017-01-01

    Salicylic acid is a signal molecule which activates plant defense against plant pathogens such as the soft rot enterobacterium Pectobacterium carotovorum subsp. carotovorum. The objectives of study were to determine bactericidal effects of salicylic acid on the growth of P. carotovorum subsp. carotovorum and secondly, assess chemical responses of P. carotovorum subsp. carotovorum to salicylic acid. Pectobacterium carotovorum subsp. carotovorum was grown in lysogeny broth amended with salicylic acid at concentrations of 0, 100, 200, 400, 800 and 1200 mg L-1. The P. carotovorum subsp. carotovorum cultures were incubated at 25°C and sampled at two time points, 0 h (sampled before incubation) and 24 h. Bacterial counts were done at the onset of the incubation (0 h) and after the 24 h incubation. The set which was incubated for 24 h was split into two, one subset was centrifuged and the other was not. From the centrifuged subset the supernatant was recovered and was, together with all the other samples (0 and 24 h not centrifuged), analyzed with1H nuclear magnetic resonance and gas chromatography. Bacterial counts done before and after incubation showed that the lower concentrations of salicylic acid, 0, 100, 200 and 400 mg L-1, supported the growth of P. carotovorum subsp. carotovorum whereas the higher concentrations of 800 and 1200 mg L-1 inhibited the growth of the bacterium completely. Nuclear magnetic resonance results showed either slight or no differences in the metabolite profiles and gas chromatography showed different responses without a clearly defined pattern among the experimental treatments. However, methanethiol was detected by both nuclear magnetic resonance and gas chromatography in all the treatments and was probably formed as a result of the breakdown of lysogeny broth. From the results obtained it was concluded that salicylic acid promotes the growth of P. carotovorum subsp. carotovorum at lower concentrations of 0-400 mg L-1 but higher

  3. Perlakuan Panas Kering dan Bakterisida untuk Menekan Infeksi Pantoea stewartii subsp. stewartii pada Benih Jagung Manis

    Directory of Open Access Journals (Sweden)

    Suswi Nalis

    2015-09-01

    Full Text Available Stewart’s Wilt is an important bacterial disease of sweet corn caused by Pantoea stewartii subsp. stewartii (synonim Erwinia stewartii. This bacteria is a seed transmitted pathogen therefore seed treatment is one method to control stewart’s wilt. The aim of this research was to study the effectiveness of dry heat, bactericide treatment, and their combinations to eliminate P. stewartii subsp. stewartii infection on sweet corn seed without damaging seed quality. The research was conducted in 3 experiments. Experiment I was conducted to determine the treatment window of dry heat and bactericide treatment. The treatment was carried out on sweet corn seed using the P. stewartii subsp. stewartii in vitro. Experiment II was conducted to study dry heat and bactericide treatment on sweet corn seed infested by P. stewartii subsp. stewartii. Experiment III was conducted to study combination of dry heat and bactericide treatment on sweet corn seed infested by P. stewartii subsp. stewartii. The results showed that dry heat treatment at 50 °C for 24 hours was able to eliminate pathogen populations in vitro but was unable to eliminate the 128 pathogen on infected seed (in vivo. Germination tests indicated that seed treatments with dry heat up to 55 °C did not decrease the germination level. The use of bactericide treatment in 100 ppm could reduce the population of bacteria on sweet corn seeds. Bactericide concentration of 150 and 200 ppm could decrease the population of bacteria on sweet corn seeds, however it could cause phytotoxic effect. The combination of bactericide (100 ppm, w/v with dry heat treatment (55 °C for 24 hours was able to eliminate bacteria on infected seed with seed germination above 85%.

  4. Complete genome and comparative analysis of Streptococcus gallolyticus subsp. gallolyticus, an emerging pathogen of infective endocarditis

    Directory of Open Access Journals (Sweden)

    Dreier Jens

    2011-08-01

    Full Text Available Abstract Background Streptococcus gallolyticus subsp. gallolyticus is an important causative agent of infectious endocarditis, while the pathogenicity of this species is widely unclear. To gain insight into the pathomechanisms and the underlying genetic elements for lateral gene transfer, we sequenced the entire genome of this pathogen. Results We sequenced the whole genome of S. gallolyticus subsp. gallolyticus strain ATCC BAA-2069, consisting of a 2,356,444 bp circular DNA molecule with a G+C-content of 37.65% and a novel 20,765 bp plasmid designated as pSGG1. Bioinformatic analysis predicted 2,309 ORFs and the presence of 80 tRNAs and 21 rRNAs in the chromosome. Furthermore, 21 ORFs were detected on the plasmid pSGG1, including tetracycline resistance genes telL and tet(O/W/32/O. Screening of 41 S. gallolyticus subsp. gallolyticus isolates revealed one plasmid (pSGG2 homologous to pSGG1. We further predicted 21 surface proteins containing the cell wall-sorting motif LPxTG, which were shown to play a functional role in the adhesion of bacteria to host cells. In addition, we performed a whole genome comparison to the recently sequenced S. gallolyticus subsp. gallolyticus strain UCN34, revealing significant differences. Conclusions The analysis of the whole genome sequence of S. gallolyticus subsp. gallolyticus promotes understanding of genetic factors concerning the pathogenesis and adhesion to ECM of this pathogen. For the first time we detected the presence of the mobilizable pSGG1 plasmid, which may play a functional role in lateral gene transfer and promote a selective advantage due to a tetracycline resistance.

  5. Human Treponema pallidum 11q/j isolate belongs to subsp. endemicum but contains two loci with a sequence in TP0548 and TP0488 similar to subsp. pertenue and subsp. pallidum, respectively.

    Directory of Open Access Journals (Sweden)

    Lenka Mikalová

    2017-03-01

    Full Text Available Treponema pallidum subsp. endemicum (TEN is the causative agent of endemic syphilis (bejel. An unusual human TEN 11q/j isolate was obtained from a syphilis-like primary genital lesion from a patient that returned to France from Pakistan.The TEN 11q/j isolate was characterized using nested PCR followed by Sanger sequencing and/or direct Illumina sequencing. Altogether, 44 chromosomal regions were analyzed. Overall, the 11q/j isolate clustered with TEN strains Bosnia A and Iraq B as expected from previous TEN classification of the 11q/j isolate. However, the 11q/j sequence in a 505 bp-long region at the TP0488 locus was similar to Treponema pallidum subsp. pallidum (TPA strains, but not to TEN Bosnia A and Iraq B sequences, suggesting a recombination event at this locus. Similarly, the 11q/j sequence in a 613 bp-long region at the TP0548 locus was similar to Treponema pallidum subsp. pertenue (TPE strains, but not to TEN sequences.A detailed analysis of two recombinant loci found in the 11q/j clinical isolate revealed that the recombination event occurred just once, in the TP0488, with the donor sequence originating from a TPA strain. Since TEN Bosnia A and Iraq B were found to contain TPA-like sequences at the TP0548 locus, the recombination at TP0548 took place in a treponeme that was an ancestor to both TEN Bosnia A and Iraq B. The sequence of 11q/j isolate in TP0548 represents an ancestral TEN sequence that is similar to yaws-causing treponemes. In addition to the importance of the 11q/j isolate for reconstruction of the TEN phylogeny, this case emphasizes the possible role of TEN strains in development of syphilis-like lesions.

  6. Human Treponema pallidum 11q/j isolate belongs to subsp. endemicum but contains two loci with a sequence in TP0548 and TP0488 similar to subsp. pertenue and subsp. pallidum, respectively.

    Science.gov (United States)

    Mikalová, Lenka; Strouhal, Michal; Oppelt, Jan; Grange, Philippe Alain; Janier, Michel; Benhaddou, Nadjet; Dupin, Nicolas; Šmajs, David

    2017-03-01

    Treponema pallidum subsp. endemicum (TEN) is the causative agent of endemic syphilis (bejel). An unusual human TEN 11q/j isolate was obtained from a syphilis-like primary genital lesion from a patient that returned to France from Pakistan. The TEN 11q/j isolate was characterized using nested PCR followed by Sanger sequencing and/or direct Illumina sequencing. Altogether, 44 chromosomal regions were analyzed. Overall, the 11q/j isolate clustered with TEN strains Bosnia A and Iraq B as expected from previous TEN classification of the 11q/j isolate. However, the 11q/j sequence in a 505 bp-long region at the TP0488 locus was similar to Treponema pallidum subsp. pallidum (TPA) strains, but not to TEN Bosnia A and Iraq B sequences, suggesting a recombination event at this locus. Similarly, the 11q/j sequence in a 613 bp-long region at the TP0548 locus was similar to Treponema pallidum subsp. pertenue (TPE) strains, but not to TEN sequences. A detailed analysis of two recombinant loci found in the 11q/j clinical isolate revealed that the recombination event occurred just once, in the TP0488, with the donor sequence originating from a TPA strain. Since TEN Bosnia A and Iraq B were found to contain TPA-like sequences at the TP0548 locus, the recombination at TP0548 took place in a treponeme that was an ancestor to both TEN Bosnia A and Iraq B. The sequence of 11q/j isolate in TP0548 represents an ancestral TEN sequence that is similar to yaws-causing treponemes. In addition to the importance of the 11q/j isolate for reconstruction of the TEN phylogeny, this case emphasizes the possible role of TEN strains in development of syphilis-like lesions.

  7. A case of acute diarrhea due to the emerging pathogen Campylobacter jejuni subsp. doylei in Southern Chile Um caso de diarréia aguda devido ao patógeno emergente Campylobacter jejuni subsp. doylei no sul do Chile

    Directory of Open Access Journals (Sweden)

    Heriberto Fernández

    2003-04-01

    Full Text Available The first documented case of acute diarrhea due to C. jejuni subsp. doylei in Chile is reported. The clinical findings, the absence of other enteropathogens, virus or parasites and the fact that C. jejuni subsp. doylei was the only bacteria isolated support the assumption that it was the etiological agent of this diarrheal case.O primeiro caso documentado de diarréia aguda por C. jejuni subsp. doylei no sul do Chile é apresentado. As características clínicas, a ausência de outros enteropatógenos, vírus ou parasitas, e o fato de C. jejuni subsp. doylei ter sido a única bactéria isolada, permitem assumir que este microrganismo é o agente etiológico neste caso de diarréia.

  8. Transcriptome-based characterization of interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus in lactose-grown chemostat cocultures

    NARCIS (Netherlands)

    Mendes, F.; Sieuwerts, S.; Hulster, de E.; Almering, M.J.; Luttik, M.A.H.; Pronk, J.T.; Smid, E.J.; Baron, P.A.; Daran-Lapujade, P.

    2013-01-01

    Mixed populations of Saccharomyces cerevisiae yeasts and lactic acid bacteria occur in many dairy, food, and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp.

  9. Transcriptome-Based Characterization of Interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus in Lactose-Grown Chemostat Cocultures

    NARCIS (Netherlands)

    Mendes, F.; Sieuwerts, S.; De Hulster, E.; Almering, M.J.; Luttik, M.A.; Pronk, J.T.; Smid, E.J.; Bron, P.A.; Daran-Lapujade, P.

    2013-01-01

    Mixed populations of Saccharomyces cerevisiae yeasts and lactic acid bacteria occur in many dairy, food, and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp.

  10. Drug susceptibility testing of Mycobacterium Avium subsp. Avium isolates from naturally infected domestic pigeons to avian tuberculosis

    Directory of Open Access Journals (Sweden)

    Kaveh Parvandar

    2016-01-01

    Conclusion: We suggest drug susceptibility testing for more nontuberculous mycobateria, particularly M. avium complex isolated from infected birds and humans, as well as molecular basics of drug sensitivity in order to detect resistance genes of pathogenic M. avium subsp. avium.

  11. Evaluation of transport and storage techniques for isolation of Campylobacter fetus subsp. jejuni from turkey cecal specimens.

    OpenAIRE

    Luechtefeld, N W; Wang, W L; Blaser, M J; Reller, L B

    1981-01-01

    Immediate culturing of fecal specimens is not always possible, and appropriate methods for transport and storage of Campylobacter fetus subsp. jejuni specimens have not been fully evaluated. Using nine techniques, we studied the survival of C. fetus subsp. jejuni in cecal specimens from infected turkeys. The organisms survived in specimens held without transport medium for 3 to 15 days (median, 9 days) at 4 degrees C, and 2 to 9 days (median, 4 days) at 25 degrees C. Only 20% of specimens fro...

  12. Key Impact of an Uncommon Plasmid on Bacillus amyloliquefaciens subsp. plantarum S499 Developmental Traits and Lipopeptide Production

    OpenAIRE

    Molinatto, Giulia; Franzil, Laurent; Steels, S?bastien; Puopolo, Gerardo; Pertot, Ilaria; Ongena, Marc

    2017-01-01

    The rhizobacterium Bacillus amyloliquefaciens subsp. plantarum S499 (S499) is particularly efficient in terms of the production of cyclic lipopeptides, which are responsible for the high level of plant disease protection provided by this strain. Sequencing of the S499 genome has highlighted genetic differences and similarities with the closely related rhizobacterium B. amyloliquefaciens subsp. plantarum FZB42 (FZB42). More specifically, a rare 8008 bp plasmid (pS499) harboring a rap-phr casse...

  13. Transcriptome-based characterization of interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus in lactose-grown chemostat cocultures.

    Science.gov (United States)

    Mendes, Filipa; Sieuwerts, Sander; de Hulster, Erik; Almering, Marinka J H; Luttik, Marijke A H; Pronk, Jack T; Smid, Eddy J; Bron, Peter A; Daran-Lapujade, Pascale

    2013-10-01

    Mixed populations of Saccharomyces cerevisiae yeasts and lactic acid bacteria occur in many dairy, food, and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus, two microorganisms that co-occur in kefir fermentations, were studied during anaerobic growth on lactose. By combining physiological and transcriptome analysis of the two strains in the cocultures, five mechanisms of interaction were identified. (i) Lb. delbrueckii subsp. bulgaricus hydrolyzes lactose, which cannot be metabolized by S. cerevisiae, to galactose and glucose. Subsequently, galactose, which cannot be metabolized by Lb. delbrueckii subsp. bulgaricus, is excreted and provides a carbon source for yeast. (ii) In pure cultures, Lb. delbrueckii subsp. bulgaricus grows only in the presence of increased CO2 concentrations. In anaerobic mixed cultures, the yeast provides this CO2 via alcoholic fermentation. (iii) Analysis of amino acid consumption from the defined medium indicated that S. cerevisiae supplied alanine to the bacterium. (iv) A mild but significant low-iron response in the yeast transcriptome, identified by DNA microarray analysis, was consistent with the chelation of iron by the lactate produced by Lb. delbrueckii subsp. bulgaricus. (v) Transcriptome analysis of Lb. delbrueckii subsp. bulgaricus in mixed cultures showed an overrepresentation of transcripts involved in lipid metabolism, suggesting either a competition of the two microorganisms for fatty acids or a response to the ethanol produced by S. cerevisiae. This study demonstrates that chemostat-based transcriptome analysis is a powerful tool to investigate microbial interactions in mixed populations.

  14. POTENSI BAKTERI ENDOFIT DALAM MENEKAN PENYAKIT LAYU STEWART (PANTOEA STEWARTII SUBSP. STEWARTII PADA TANAMAN JAGUNG

    Directory of Open Access Journals (Sweden)

    Haliatur Rahma

    2017-04-01

    Full Text Available Potential of endophytic bacteria to control stewart wilt disease (Pantoea stewartii subsp. stewartii in maize. The purpose of this study was to explore endophytic bacteria from seedling, maize roots and grass roots as well as to test the ability of endophytic bacteria which could potentially suppress stewart wilt disease development in maize. Characterization of endophytic bacteria as biocontrol agents including: do not induce HR on tobacco, synthesize IAA, dissolve phosphate, produce siderophores, and antibiotic to Pantoea stewartii subsp. stewartii (Pnss. The results of research shoed 17 isolates of endophytic bacteria potentially as candidate biocontrol agents. Nine isolates were able to produce IAA, siderofores and phosphatase; two isolates produce IAA and phosphatase; six isolates produce IAA. Six isolates ie: AR1, AJ34, AJ15, AJ19, and AJ14 AN6, can increase maize plant resistance and suppress stewart wilt disease severity with a range of 48.95-55.60%.

  15. Anatomy and Micromorphology of Inula helenium subsp. orgyalis and I. ensifolia (Asteraceae from Turkey

    Directory of Open Access Journals (Sweden)

    Tulay AYTAS AKCIN

    2017-03-01

    Full Text Available Inula helenium L. subsp. orgyalis (Boiss. Grierson and Inula ensifolia L. were investigated anatomically and micromorphologically. The secretory cavities in the leaves and stem of both investigated taxa were located in the neighbourhood of the vascular bundles and in the rhizomes in the secondary cortex. The leaf mesophylls of investigated Inula taxa were homogeneous. Stomata were anomocytic in two species. The distribution and density of the eglandular and glandular trichomes provide information of taxonomical significance. Moreover, the cypselas of I. helenium L. subsp. orgyalis were homomorphic, whereas in I. ensifolia cypselas were heteromorphic. Additionally, the number of ribs, the shape of carpopodium and stylopodium were diagnostic taxonomic characters between the two taxa.

  16. Occurrence of Clavibacter michiganensis subsp. michiganensis, the causal agent of bacterial canker of tomato, in Syria

    Directory of Open Access Journals (Sweden)

    Radwan FTAYEH

    2010-09-01

    Full Text Available Several surveys were carried out to evaluate the occurrence of bacterial canker of tomato caused by Clavibacter michiganensis subsp. michiganensis (Cmm in Syria, especially in the North-West provinces Latakiaand Tartous. The surveys revealed typical disease symptoms in greenhouses where the tomato cvs. Dima, Huda and Astona were grown, such as dark brown to black lesions on the leaf margins, wilting of whole plants, stunting, and vascular discoloration. The disease incidence in such greenhouses was 15% in the spring of 2007, and up to 70% by the end of July. Ten isolates obtained from diseased plants at different locations in these two provinces were identified as Clavibacter michiganensis subsp. michiganensis using classical microbiological tests as well as PCR. This is the first detailed proof of the occurrence of bacterial canker of tomato in Syria.

  17. Acacia tortilis subsp. heteracantha productivity in the Tugela Dry Valley Bushveld: preliminary results

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    S. J. Milton

    1983-11-01

    Full Text Available Acacia tortilis Hayne subsp.  heteracantha (Burch. Brenan dominates secondary succession in the Tugela Dry Valley Bushveld of the Natal midlands. The parts of KwaZulu in this veld type are impoverished, overpopulated and over-grazed. Preliminary results indicate that at a density of 2 700 ± 600 trees/ha there is a standing crop of c.2,87 t/ha (DM of acacia twigs suitable for hand pruning and milling into fodder, but that this is a costly process. Herbage biomass peaked at 0,73 t/ha (DM. Veld condition assessments suggested a stocking rate of|0,l AU/ha (grazers, but actual grazer stocking rates may be many times this density. It is recommended that the browser/grazer ratio be altered to make use of the c. 1,05 t/ha (DM of shoot growth produced annually by A. tortilis subsp.  heteracantha.

  18. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: Characterization of the bacteriocin

    Science.gov (United States)

    Furtado, Danielle N.; Todorov, Svetoslav D.; Landgraf, Mariza; Destro, Maria T.; Franco, Bernadette D.G.M.

    2014-01-01

    Lactic acid bacteria capable of producing bacteriocins and presenting probiotic potential open innovative technological applications in the dairy industry. In this study, a bacteriocinogenic strain (Lactococcus lactis subsp. lactis DF4Mi) was isolated from goat milk, and studied for its antimicrobial activity. The bacteriocin presented a broad spectrum of activity, was sensitive to proteolytic enzymes, resistant to heat and pH extremes, and not affected by the presence of SDS, Tween 20, Tween 80, EDTA or NaCl. Bacteriocin production was dependent on the components of the culture media, especially nitrogen source and salts. When tested by PCR, the bacteriocin gene presented 100% homology to nisin Z gene. These properties indicate that this L. lactis subsp. lactis DF4Mi can be used for enhancement of dairy foods safety and quality. PMID:25763065

  19. Restoring catalase activity in Staphylococcus aureus subsp. anaerobius leads to loss of pathogenicity for lambs.

    Science.gov (United States)

    de la Fuente, Ricardo; Díez, Rosa M; Domínguez-Bernal, Gustavo; Orden, José A; Martínez-Pulgarín, Susana

    2010-01-01

    Staphylococcus aureus subsp. anaerobius, a microaerophilic and catalase-negative bacterium, is the etiological agent of abscess disease, a specific chronic condition of sheep and goats, which is characterized by formation of necrotic lesions that are located typically in superficial lymph nodes. We constructed an isogenic mutant of S. aureus subsp. anaerobius (RDKA84) that carried a repaired and functional catalase gene from S. aureus ATCC 12600, to investigate whether the lack of catalase in S. aureus subsp. anaerobius plays a role in its physiological and pathogenic characteristics. The catalase activity had no apparent influence on the in vitro growth characteristics of RDKA84, which, like the wild-type, did not grow on aerobically incubated agar plates. Restoration of catalase activity in RDKA84 substantially increased resistance to H2O2 when analyzed in a death assay. The intracellular survival rates of the catalase-positive mutant RDKA84 in polymorphonuclear neutrophils (PMN) isolated from adult sheep were significantly higher than those of the wild-type, while no differences were found with PMN isolated from lambs. RDKA84 showed significantly lower survival rates in murine macrophages (J774A.1 cells) than the wild-type strains did, whereas, in bovine mammary epithelial cells (MAC-T), no differences in intracellular survival were observed. Interestingly, the virulence for lambs, the natural host for abscess disease, of the catalase-positive mutant RDKA84 was reduced dramatically in comparison with wild-type S. aureus subsp. anaerobius in two experimental models of infection. Copyright (c) INRA, EDP Sciences, 2010.

  20. FUNCTIONAL DIOECY AND MOTH POLLINATION IN CABRALEA CANJERANA SUBSP. CANJERANA (MELIACEAE

    Directory of Open Access Journals (Sweden)

    Edivani V. Franceschinelli

    2015-01-01

    Full Text Available Cabralea canjerana subsp . canjerana es una especie común en la Mata Atlántica del sudeste de Bra - sil ; sin embargo, poco se sabe acerca de sus estrategias reproductivas . Este estudio tiene como objetivo entender la biología reproductiva de esta subespecie , incluyendo su biología floral, sistema sexual y la biología de la polinización. Las plantas masculinas y femeninas tienen flores morfológicamente simi - lares, pero las plantas masculinas tienen inflorescencias más grandes con más flores que las femeninas. Además, las flores femeninas permanecen abiertas y receptivas por dos días, mientras que las flores masculinas duran sólo un día, compensando así el desequilibrio de la cantidad de flores por planta entre los dos sexos. Flores masculinas y femeninas semejantes , como se observan en C. canjerana subsp . canjerana , son poco comunes entre las especies dioicas . La alta frecuencia de las polillas que visitan las flores , el periodo de apertura de la flor y de la producción de néctar sugieren que las polillas son polini - zadores de esta subespecie. Contrastando con el dimorfismo floral encontrado en C. canjerana subsp . polytricha , las variaciones en la morfología floral no están relacionadas con el sexo de la planta en la subsp . canjerana . Este y otros resultados encontrados en este estudio sugieren que estas dos subspecies de C. canjerana podrían ser especies distintas.

  1. Molecular biology techniques as a tool for detection and characterisation of Mycobacterium avium subsp. paratuberculosis

    OpenAIRE

    Englund, Stina

    2002-01-01

    Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) is the causative agent of paratuberculosis, also known as Johne’s disease, a chronic intestinal infection in cattle and other ruminants. Paratuberculosis is characterised by diarrhea and weight loss that occurs after a period of a few months up to several years without any clinical signs. The considerable economic losses to dairy and beef cattle producers are caused by reduced milk production and poor reproduction performance i...

  2. MORPHOLOGICAL AND ANATOMICAL STUDY ON ENDEMIC CROCUS OLIVIERI GAY SUBSP. ISTANBULENSIS MATHEW SUBSPECIES (IRIDACEAE

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    Kadriye Yetişen

    2013-02-01

    Full Text Available In this study, morphological and anatomical properties of Crocus olivieri Gay subsp. istanbulensis Mathew were investigated. Cross-sections of root, scape and leaf parts of the plant were examined anddemonstrated by photographs. Most of the anatomical properties are similar to the other member of Iridaceae family. Sclerenchyma groups were observed around to leaf vascular bundle. Morphological and anatomical findings compared with other two subspecies of Crocus olivieri.

  3. Inactivation of Xanthomonas citri subsp. citri and Effect on Infection of Citrus Canker by Gamma Irradiation

    OpenAIRE

    Kyung Nam Kim; Min A Song; Yong Chull Jeun; Sang Heon Han; Seong Joon Song

    2015-01-01

    Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) has been quarantined by many countries in the world. Recently, the usage of methyl bromide should be limited, application by gamma irradiation on the agricultural production is raised as an alternative method. In this study, the level of gamma irradiation which could decrease of population of Xcc in the suspension or on the surface of citrus fruit was investigated. The D10 value of Xcc, which is radiation dose required t...

  4. Susceptibility of Campylobacter Fetus Subsp. Jejuni, Isolated from Patients in Jakarta, Indonesia to Ten Antimicrobial Agents

    Science.gov (United States)

    1981-06-16

    antimicrobials was tested against 28 Campylohacter letus subsp. jejuni isolates cultured from the stools of human gastroenteritis and suspected typhoid fever...isolated from the faeces of gastroenteritis and suspected typhoid fever patients in Jakarta, Indonesia and to compare the MIC values with those...jejuni strains used in this study were cultured from the faeces of 19 gastroenteritis and five suspected typhoid fever patients examined at the

  5. Insights into physiological traits of Bifidobacterium animalis subsp. lactis BB-12 through membrane proteome analysis

    DEFF Research Database (Denmark)

    Gilad, Ofir; Hjernø, Karin; Østerlund, Eva Christina

    2012-01-01

    Bifidobacterium animalis subsp. lactis BB-12 is a widely used probiotic strain associated with a variety of health-promoting traits. There is, however, only limited knowledge available regarding the membrane proteome and the proteins involved in oligosaccharide transport in BB-12. We applied two ......, phosphate or exopolysaccharides, or to belong to the F1F0-ATP-synthetase complex and the protein translocation machinery, respectively....

  6. Stress resistance of biofilm and planktonic Lactobacillus plantarum subsp. plantarum JCM 1149

    OpenAIRE

    Kubota, Hiromi; Senda, Shouko; Tokuda, Hajime; Uchiyama, Hiroo; Nomura, Nobuhiko

    2009-01-01

    The purpose of this study was to investigate the change in resistance of biofilm and planktonic food spoilage lactic acid bacteria (LAB) to environmental stresses, which strongly inhibit bacterial growth and are important in food preservation or in disinfection. The stress responses of biofilm and planktonic cells of Lactobacillus plantarum subsp. plantarum JCM 1149, which was used as a model spoilage bacterium, in various organic acids (namely, acetic acid, citric acid, lactic acid, and mali...

  7. Molecular Characterization of Multidrug-Resistant Salmonella enterica subsp. enterica Serovar Typhimurium Isolates from Swine

    OpenAIRE

    Gebreyes, Wondwossen Abebe; Altier, Craig

    2002-01-01

    As part of a longitudinal study of antimicrobial resistance among salmonellae isolated from swine, we studied 484 Salmonella enterica subsp. enterica serovar Typhimurium (including serovar Typhimurium var. Copenhagen) isolates. We found two common pentaresistant phenotypes. The first was resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (the AmCmStSuTe phenotype; 36.2% of all isolates), mainly of the definitive type 104 (DT104) phage type (180 of 187 ...

  8. Volatile Constituents of Ferula communis L. subsp. communis Growing Spontaneously in Greece

    Directory of Open Access Journals (Sweden)

    Stavroula Manolakou

    2013-01-01

    Full Text Available The essential oils of Greek Ferula communis subsp. communis from different plant parts were obtained by hydrodistillation and analyzed by means of GC and GC-MS. Ninety three compounds were identified in the total essential oils. Sesqui terpenes were the most dominant class of compounds in the leaves and inflorescences oils, while infructescences oils were rich in monoterpenes with α-pinene (35.2-40.6% being the dominant component.

  9. ADHESION AND PHAGOCYTOSIS PROPERTIES OFSTREPTOCOCCUS EQUI SUBSP. ZOOEPIDEMICUS ISOLATES FROM INDONESIA

    OpenAIRE

    Salasia, Siti Isrina Oktavia

    2015-01-01

    An infectious diseases outbreak affecting pigs and monkeys in Bali in 1994, with case fatality rate (CFR) 75%. The disease widespread rapidly to the other district in Indonesia and developed serious problems. The causative agent involving the disease was Streptococcus equi subsp. zooepidemicus, a β-hemoIytic type of the streptococcus. The disease was consistently associated with meningitis, therefore the disease was named Streprococcal meningitis. The main aim of this study is todetect factor...

  10. Penggunaan Kultur Makrofag untuk Pengujian Virulensi Streptococcus equi subs. Zooepidemicus (THE USE OF MACROPHAGE CULTURE IN VIRULENCE ASSAY OF STREPTOCOCCUS EQUI SUBSP. ZOOEPIDEMICUS

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    Iwan Harjono Utama

    2016-08-01

    Full Text Available Penggunaan Kultur Makrofag untuk Pengujian Virulensi Streptococcus equi subs. Zooepidemicus (THE USE OF MACROPHAGE CULTURE IN VIRULENCE ASSAY OF STREPTOCOCCUS EQUI SUBSP. ZOOEPIDEMICUS

  11. Listeria innocua and Lactobacillus delbrueckii subsp. bulgaricus employ different strategies to cope with acid stress

    DEFF Research Database (Denmark)

    Shabala, Lana; McMeekin, Tom; Budde, Birgitte Bjørn

    2006-01-01

    Responses of Listeria innocua and Lactobacillus delbrueckii subsp. bulgaricus to a rapid change in extracellular pH (pHex) from pHex 6 to a range of concentrations down to pHex 3.0 were examined, using HCl and lactic acid (LA) as acidulants. A new fluorescent probe 5-(and-6)-carboxy-2', 7...... technique (MIFE), respectively. L. innocua maintained a relatively constant pHi of 5.5-6.1 at pHex 4 and 5 via H+ extrusion. In contrast, L. delbrueckii subsp. bulgaricus progressively lowered pHi towards pHex over the entire pHex range examined. The type of acidulant used influenced pH regulation with both...... pHi and H+ -fluxes being more severely affected by LA compared to HCl. Overall, our data demonstrated different adaptive strategies in these two bacteria. While L. innocua expels protons to maintain a constant pHi, L. delbrueckii subsp. bulgaricus allows proton entry after acidic treatment so that p...

  12. Lactobacillus delbrueckii subsp. bulgaricus CRL 454 cleaves allergenic peptides of β-lactoglobulin.

    Science.gov (United States)

    Pescuma, Micaela; Hébert, Elvira M; Haertlé, Thomas; Chobert, Jean-Marc; Mozzi, Fernanda; Font de Valdez, Graciela

    2015-03-01

    Whey, a cheese by-product used as a food additive, is produced worldwide at 40.7 million tons per year. β-Lactoglobulin (BLG), the main whey protein, is poorly digested and is highly allergenic. We aimed to study the contribution of Lactobacillus delbrueckii subsp. bulgaricus CRL 454 to BLG digestion and to analyse its ability to degrade the main allergenic sequences of this protein. Pre-hydrolysis of BLG by L. delbrueckii subsp. bulgaricus CRL 454 increases digestion of BLG assayed by an in vitro simulated gastrointestinal system. Moreover, peptides from hydrolysis of the allergenic sequences V41-K60, Y102-R124, C121-L140 and L149-I162 were found when BLG was hydrolysed by this strain. Interestingly, peptides possessing antioxidant, ACE inhibitory, antimicrobial and immuno-modulating properties were found in BLG degraded by both the Lactobacillus strain and digestive enzymes. To conclude, pre-hydrolysis of BLG by L. delbrueckii subsp. bulgaricus CRL 454 has a positive effect on BLG digestion and could diminish allergenic reactions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Genetic variability and population structure of Disanthus cercidifolius subsp. longipes (Hamamelidaceae based on AFLP analysis.

    Directory of Open Access Journals (Sweden)

    Yi Yu

    Full Text Available Disanthus cercidifolius subsp. longipes is an endangered species in China. Genetic diversity and structure analysis of this species was investigated using amplified fragments length polymorphism (AFLP fingerprinting. Nei's gene diversity ranged from 0.1290 to 0.1394. The AMOVA indicated that 75.06% of variation was distributed within populations, while the between-group component 5.04% was smaller than the between populations-within-group component 19.90%. Significant genetic differentiation was detected between populations. Genetic and geographical distances were not correlated. PCA and genetic structure analysis showed that populations from East China were together with those of the Nanling Range. These patterns of genetic diversity and levels of genetic variation may be the result of D. c. subsp. longipes restricted to several isolated habitats and "excess flowers production, but little fruit set". It is necessary to protect all existing populations of D. c. subsp. longipes in order to preserve as much genetic variation as possible.

  14. Taxonomy, distribution and diversity of Ficus palmata Forssk. Subsp. virgata (Roxb. Browicz (Moraceae in India

    Directory of Open Access Journals (Sweden)

    R. Tiwari

    2014-08-01

    Full Text Available The examination of a large number of herbarium specimens combined with field observations reveal that Ficus palmata Forssk. subsp. palmata does not occur in India. The Indian plants occurring in the wild from the north-west to the south belong to F. palmata Forssk. subsp. virgata (Roxb. Browicz. The maximum concentration of the taxon lies in northern India extending up to about 2200m altitude in the Himalaya. In southern India, the taxon is reported only in Andhra Pradesh. F. palmata subsp. virgata is notoriously variable in its entire range of distribution in almost all morphological characters. The variations are continuous and its two extreme forms, with entire leaves and lobed leaves, are connected with numerous intermediate forms. The taxon is closely allied to F. carica L., which is distributed from the Mediterranean region to Afghanistan and occurs only in cultivation in some parts of India. The paper also explains the relationship of the taxon with its closely allied species and provides a key to discriminate among them. In this paper, the taxon is described, illustrated with colour photographs and line drawings and provided with a distribution map.

  15. Identification and characterization of Nip, necrosis-inducing virulence protein of Erwinia carotovora subsp. carotovora.

    Science.gov (United States)

    Mattinen, Laura; Tshuikina, Marina; Mäe, Andres; Pirhonen, Minna

    2004-12-01

    Erwinia carotovora subsp. carotovora is a gram-negative bacterium that causes soft rot disease of many cultivated crops. When a collection of E. carotovora subsp. carotovora isolates was analyzed on a Southern blot using the harpin-encoding gene hrpN as probe, several harpinless isolates were found. Regulation of virulence determinants in one of these, strain SCC3193, has been characterized extensively. It is fully virulent on potato and in Arabidopsis thaliana. An RpoS (SigmaS) mutant of SCC3193, producing elevated levels of secreted proteins, was found to cause lesions resembling the hypersensitive response when infiltrated into tobacco leaf tissue. This phenotype was evident only when bacterial cells had been cultivated on solid minimal medium at low pH and temperature. The protein causing'the cell death was purified and sequenced, and the corresponding gene was cloned. The deduced sequence of the necrosis-inducing protein (Nip) showed homology to necrosis- and ethylene-inducing elicitors of fungi and oomycetes. A mutant strain of E. carotovora subsp. carotovora lacking the nip gene showed reduced virulence in potato tuber assay but was unaffected in virulence in potato stem or on other tested host plants.

  16. Identification of an Extracellular Endoglucanase That Is Required for Full Virulence in Xanthomonas citri subsp. citri.

    Directory of Open Access Journals (Sweden)

    Tian Xia

    Full Text Available Xanthomonas citri subsp. citri causes citrus canker disease, which is characterized by the formation of water-soaked lesions, white or yellow spongy pustules and brown corky canker. In this work, we report the contribution of extracellular endoglucanase to canker development during infection. The ectopic expression of nine putative cellulases in Escherichia coli indicated that two endoglucanases, BglC3 and EngXCA, show carboxymethyl cellulase activity. Both bglC3 and engXCA genes were transcribed in X. citri subsp. citri, however, only BglC3 protein was detected outside the cell in western blot analysis. The deletion of bglC3 gene resulted in complete loss of extracellular carboxymethyl cellulase activity and delayed the onset of canker symptoms in both infiltration- and wound-inoculation assays. When growing in plant tissue, the cell density of bglC3 mutant was lower than that of the wild type. Our data demonstrated that BglC3 is an extracellular endoglucanase required for the full virulence of X. citri subsp. citri.

  17. Geography of Genetic Structure in Barley Wild Relative Hordeum vulgare subsp. spontaneum in Jordan.

    Science.gov (United States)

    Thormann, Imke; Reeves, Patrick; Reilley, Ann; Engels, Johannes M M; Lohwasser, Ulrike; Börner, Andreas; Pillen, Klaus; Richards, Christopher M

    2016-01-01

    Informed collecting, conservation, monitoring and utilization of genetic diversity requires knowledge of the distribution and structure of the variation occurring in a species. Hordeum vulgare subsp. spontaneum (K. Koch) Thell., a primary wild relative of barley, is an important source of genetic diversity for barley improvement and co-occurs with the domesticate within the center of origin. We studied the current distribution of genetic diversity and population structure in H. vulgare subsp. spontaneum in Jordan and investigated whether it is correlated with either spatial or climatic variation inferred from publically available climate layers commonly used in conservation and ecogeographical studies. The genetic structure of 32 populations collected in 2012 was analyzed with 37 SSRs. Three distinct genetic clusters were identified. Populations were characterized by admixture and high allelic richness, and genetic diversity was concentrated in the northern part of the study area. Genetic structure, spatial location and climate were not correlated. This may point out a limitation in using large scale climatic data layers to predict genetic diversity, especially as it is applied to regional genetic resources collections in H. vulgare subsp. spontaneum.

  18. Production and characterization of bioemulsifier from a marine bacterium, Acinetobacter calcoaceticus subsp. anitratus SM7

    Directory of Open Access Journals (Sweden)

    Kulnaree Phetrong

    2008-05-01

    Full Text Available Marine bacterium strain SM7 was isolated as a bioemulsifier-producing bacterium from oil-spilled seawater in Songkhla lagoon, Thailand. It was identified as Acinetobacter calcoaceticus subsp. anitratus based on morphology, biochemicalcharacteristics and 16S rRNA sequence. A. calcoaceticus subsp. anitratus SM7 produced an extracellular emulsifying agent when grown in a minimal salt medium (pH 7.0 containing 0.3% (v/v n-heptadecane and 0.1% (w/v ammoniumhydrogen carbonate as carbon source and nitrogen source, respectively, at 30oC with agitation rate of 200 rpm. Crude bioemulsifier was recovered from the culture supernatant by ethanol precipitation with a yield of 2.94 g/l and had a criticalemulsifier concentration of 0.04 g/ml. The crude bioemulsifier was capable of emulsifying n-hexadecane in a broad pH range (6-12, temperatures (30-121oC and in the presence of NaCl up to 12% (w/v. The bioemulsifier was stable in saltsolution ranging from 0 to 0.1% (w/v of MgCl2 and CaCl2. The broad range of pH stability, thermostability and salt tolerance suggested that the bioemulsifier from A. calcoaceticus subsp. anitratus SM7 could be useful in environmentalapplication, especially bioremediation of oil-polluted seawater.

  19. OmpF of Pectobacterium carotovorum subsp. carotovorum Pcc3 is required for carocin D sensitivity.

    Science.gov (United States)

    Lim, Jeong-A; Hong, Jisoo; Kim, Jonguk; Heu, Sunggi; Roh, Eunjung

    2016-12-01

    Carocin D is a bacteriocin produced by Pectobacterium carotovorum subsp. carotovorum Pcc21. Carocin D inhibits the growth of P carotovorum subsp. carotovorum and closely related strains. Pectobacterium carotovorum subsp. carotovorum is a causative bacterium for soft rot disease and leads to severe economic losses. Bacteriocins recognize and interact with a specific membrane protein of target bacteria as a receptor. To identify the receptor responsible for carocin D recognition, mutants that underwent a phenotypic change from carocin D sensitivity to carocin D insensitivity were screened. Based on Tn5 insertions, carocin D sensitivity was dependent on expression of the outer membrane protein OmpF. The insensitivity of the mutant (Pcc3MR) to carocin D was complemented with ompF from carocin D-sensitive strains, not from carocin D-resistant strains. The selectivity between sensitive and resistant strains could be attributed to variation in OmpFs in the cell-surface-exposed regions. Based on sequence analysis and complementation assays, it appears that carocin D uses OmpF as a receptor and is translocated by the TonB system. According to previously reported translocation mechanisms of colicins, OmpF works along with the TolA system rather than the TonB system. Therefore, the current findings suggest that carocin D is imported by a unique colicin-like bacteriocin translocation system. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Evaluation of Lavandula stoechas L. subsp. stoechas L., Mentha spicata L. subsp. spicata L. essential oils and their main components against sinusitis pathogens.

    Science.gov (United States)

    Karaca, Nursenem; Demirci, Betül; Demirci, Fatih

    2018-02-27

    Lavandula stoechas subsp. stoechas and Mentha spicata subsp. spicata are used for the treatment of sinusitis in Turkish folk medicine. The components of essential oils obtained by hydrodistillation were determined by gas chromatography-flame ionization detector (GC-FID), gas chromatography/mass spectrometry (GC/MS), and thin layer chromatography (TLC). Major components of L. stoechas and M. spicata oils were determined as camphor (46.7%) and carvone (60.6%), respectively. The antibacterial activity of essential oils and their main components were tested against the common selected sinusitis pathogens Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, and Pseudomonas aeruginosa using in vitro agar diffusion, microdilution, and vapor diffusion methods. As a result, the tested plant materials, which are locally and natively used against sinusitis, were relatively mild antibacterial (in vitro MICs 310-1250 μg/mL) in action. To use essential oils and their components safely in sinusitis therapy, further detailed in vivo experiments are needed to support their efficacy.

  1. Effects of Roundup(®) and glyphosate on three food microorganisms: Geotrichum candidum, Lactococcus lactis subsp. cremoris and Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Clair, Emilie; Linn, Laura; Travert, Carine; Amiel, Caroline; Séralini, Gilles-Eric; Panoff, Jean-Michel

    2012-05-01

    Use of many pesticide products poses the problem of their effects on environment and health. Amongst them, the effects of glyphosate with its adjuvants and its by-products are regularly discussed. The aim of the present study was to shed light on the real impact on biodiversity and ecosystems of Roundup(®), a major herbicide used worldwide, and the glyphosate it contains, by the study of their effects on growth and viability of microbial models, namely, on three food microorganisms (Geotrichum candidum, Lactococcus lactis subsp. cremoris and Lactobacillus delbrueckii subsp. bulgaricus) widely used as starters in traditional and industrial dairy technologies. The presented results evidence that Roundup(®) has an inhibitory effect on microbial growth and a microbicide effect at lower concentrations than those recommended in agriculture. Interestingly, glyphosate at these levels has no significant effect on the three studied microorganisms. Our work is consistent with previous studies which demonstrated that the toxic effect of glyphosate was amplified by its formulation adjuvants on different human cells and other eukaryotic models. Moreover, these results should be considered in the understanding of the loss of microbiodiversity and microbial concentration observed in raw milk for many years.

  2. Characterization of Anaplasma marginale subsp. centrale Strains by Use of msp1aS Genotyping Reveals a Wildlife Reservoir.

    Science.gov (United States)

    Khumalo, Zamantungwa T H; Catanese, Helen N; Liesching, Nicole; Hove, Paidashe; Collins, Nicola E; Chaisi, Mamohale E; Gebremedhin, Assefaw H; Oosthuizen, Marinda C; Brayton, Kelly A

    2016-10-01

    Bovine anaplasmosis caused by the intraerythrocytic rickettsial pathogen Anaplasma marginale is endemic in South Africa. Anaplasma marginale subspecies centrale also infects cattle; however, it causes a milder form of anaplasmosis and is used as a live vaccine against A. marginale There has been less interest in the epidemiology of A. marginale subsp. centrale, and, as a result, there are few reports detecting natural infections of this organism. When detected in cattle, it is often assumed that it is due to vaccination, and in most cases, it is reported as coinfection with A. marginale without characterization of the strain. A total of 380 blood samples from wild ruminant species and cattle collected from biobanks, national parks, and other regions of South Africa were used in duplex real-time PCR assays to simultaneously detect A. marginale and A. marginale subsp. centrale. PCR results indicated high occurrence of A. marginale subsp. centrale infections, ranging from 25 to 100% in national parks. Samples positive for A. marginale subsp. centrale were further characterized using the msp1aS gene, a homolog of msp1α of A. marginale, which contains repeats at the 5' ends that are useful for genotyping strains. A total of 47 Msp1aS repeats were identified, which corresponded to 32 A. marginale subsp. centrale genotypes detected in cattle, buffalo, and wildebeest. RepeatAnalyzer was used to examine strain diversity. Our results demonstrate a diversity of A. marginale subsp. centrale strains from cattle and wildlife hosts from South Africa and indicate the utility of msp1aS as a genotypic marker for A. marginale subsp. centrale strain diversity. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  3. Phenotypic, Genotypic, and Antimicrobial Characteristics of Streptococcus halichoeri Isolates from Humans, Proposal To Rename Streptococcus halichoeri as Streptococcus halichoeri subsp. halichoeri, and Description of Streptococcus halichoeri subsp. hominis subsp. nov., a Bacterium Associated with Human Clinical Infections.

    Science.gov (United States)

    Shewmaker, P L; Whitney, A M; Humrighouse, B W

    2016-03-01

    Phenotypic, genotypic, and antimicrobial characteristics of six phenotypically distinct human clinical isolates that most closely resembled the type strain of Streptococcus halichoeri isolated from a seal are presented. Sequencing of the 16S rRNA, rpoB, sodA, and recN genes; comparative whole-genome analysis; conventional biochemical and Rapid ID 32 Strep identification methods; and antimicrobial susceptibility testing were performed on the human isolates, the type strain of S. halichoeri, and type strains of closely related species. The six human clinical isolates were biochemically indistinguishable from each other and showed 100% 16S rRNA, rpoB, sodA, and recN gene sequence similarity. Comparative 16S rRNA gene sequencing analysis revealed 98.6% similarity to S. halichoeri CCUG 48324(T), 97.9% similarity to S. canis ATCC 43496(T), and 97.8% similarity to S. ictaluri ATCC BAA-1300(T). A 3,530-bp fragment of the rpoB gene was 98.8% similar to the S. halichoeri type strain, 84.6% to the S. canis type strain, and 83.8% to the S. ictaluri type strain. The S. halichoeri type strain and the human clinical isolates were susceptible to the antimicrobials tested based on CLSI guidelines for Streptococcus species viridans group with the exception of tetracycline and erythromycin. The human isolates were phenotypically distinct from the type strain isolated from a seal; comparative whole-genome sequence analysis confirmed that the human isolates were S. halichoeri. On the basis of these results, a novel subspecies, Streptococcus halichoeri subsp. hominis, is proposed for the human isolates and Streptococcus halichoeri subsp. halichoeri is proposed for the gray seal isolates. The type strain of the novel subspecies is SS1844(T) = CCUG 67100(T) = LMG 28801(T). Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  4. Type IV Pili are required for virulence, twitching motility, and biofilm formation of acidovorax avenae subsp. Citrulli.

    Science.gov (United States)

    Bahar, Ofir; Goffer, Tal; Burdman, Saul

    2009-08-01

    Acidovorax avenae subsp. citrulli is the causal agent of bacterial fruit blotch (BFB), a threatening disease of watermelon, melon, and other cucurbits. Despite the economic importance of BFB, relatively little is known about basic aspects of the pathogen's biology and the molecular basis of its interaction with host plants. To identify A. avenae subsp. citrulli genes associated with pathogenicity, we generated a transposon (Tn5) mutant library on the background of strain M6, a group I strain of A. avenae subsp. citrulli, and screened it for reduced virulence by seed-transmission assays with melon. Here, we report the identification of a Tn5 mutant with reduced virulence that is impaired in pilM, which encodes a protein involved in assembly of type IV pili (TFP). Further characterization of this mutant revealed that A. avenae subsp. citrulli requires TFP for twitching motility and wild-type levels of biofilm formation. Significant reductions in virulence and biofilm formation as well as abolishment of twitching were also observed in insertional mutants affected in other TFP genes. We also provide the first evidence that group I strains of A. avenae subsp. citrulli can colonize and move through host xylem vessels.

  5. Development of a sensitive nested PCR method for the specific detection of Mycoplasma mycoides subsp. mycoides SC.

    Science.gov (United States)

    Miserez, R; Pilloud, T; Cheng, X; Nicolet, J; Griot, C; Frey, J

    1997-04-01

    A specific and sensitive test for the detection of Mycoplasma mycoides subsp. mycoides small colony type (SC), the aetiological agent of contagious bovine pleuropneumonia (CBPP) was developed using two nested PCR reactions. The PCR reactions are based on the nucleotide sequence of lipoprotein P72 of M. mycoides subsp. mycoides SC. The two specific oligonucleotide primer pairs were chosen to match those sequence segments of the P72 gene which differ most from the gene of the closely related lipoprotein P67 of Mycoplasma sp. bovine group 7 (strain PG50). The nested PCR reacted with all of the 34 different strains of M. mycoides subsp. mycoides SC analysed, and gave no amplification product with any of the closely related mycoplasmas tested, showing its high specificity. In bronchial lavage fluid experimentally contaminated with M. mycoides subsp. mycoides SC, the assay was able to detect as few as two viable cells per ml using a simple lysis procedure prior to the amplification step. With clinical samples, the sensitivity of the nested PCR was about 10(4)-10(5) higher than that of single PCR amplifications performed under the same conditions. The assay was also successfully used to detect M. mycoides subsp. mycoides SC in bronchial lavage fluid of experimentally infected cattle and proved to be more sensitive than classical culture methods.

  6. Phenotypic variation in Lactococcus lactis subsp. lactis isolates derived from intestinal tracts of marine and freshwater fish.

    Science.gov (United States)

    Itoi, S; Yuasa, K; Washio, S; Abe, T; Ikuno, E; Sugita, H

    2009-09-01

    We compared phenotypic characteristics of Lactococcus lactis subsp. lactis derived from different sources including the intestinal tract of marine fish and freshwater fish, and cheese starter culture. In the phylogenetic analysis based on partial 16S rRNA gene nucleotide sequences (1371 bp), freshwater fish-, marine fish- and cheese starter culture-derived strains were identical to that of L. lactis subsp. lactis previously reported. Fermentation profiles determined using the API 50 CH system were similar except for fermentation of several sugars including l-arabinose, mannitol, amygdalin, saccharose, trehalose, inulin and gluconate. The strains did have distinct levels of halotolerance: marine fish-derived strains > cheese starter-derived strain > freshwater fish-derived isolate. Lactococcus lactis subsp. lactis showed extensive diversity in phenotypic adaptation to various environments. The phenotypic properties of these strains suggested that L. lactis subsp. lactis strains from fish intestine have additional functions compared with the cheese starter-derived strain that has previously described. The unique phenotypic traits of the fish intestinal tract-derived L. lactis subsp. lactis might make them useful as a probiotics in aquaculture, and contribute to the development of functional foods and novel food additives, since the strains derived from fish intestines might have additional functions such as antibacterial activity.

  7. Profiles of Volatile Flavor Compounds in Milk Fermented with Different Proportional Combinations of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus.

    Science.gov (United States)

    Dan, Tong; Wang, Dan; Wu, Shimei; Jin, Rulin; Ren, Weiyi; Sun, Tiansong

    2017-09-29

    Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus are key factors in the fermentation process and the final quality of dairy products worldwide. This study was performed to investigate the effects of the proportions of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus isolated from traditionally fermented dairy products in China and Mongolia on the profile of volatile compounds produced in samples. Six proportional combinations (1:1, 1:10, 1:50, 1:100, 1:1000, and 1:10,000) of L. delbrueckii subsp. bulgaricus IMAU20401 to S. thermophilus ND03 were considered, and the volatiles were identified and quantified by solid-phase microextraction and gas chromatography-mass spectrometry (SPME-GC-MS) against an internal standard. In total, 89 volatile flavor compounds, consisting of aldehydes, ketones, acids, alcohols, esters, and aromatic hydrocarbons, were identified. Among these, some key flavor volatile compounds were identified, including acetaldehyde, 3-methylbutanal, acetoin, 2-heptanone, acetic acid, butanoic acid, and 3-methyl-1-butanol. The of L. delbrueckii subsp. bulgaricus IMAU20401 to S. thermophilus ND03 influenced the type and concentration of volatiles produced. In particular, aldehydes and ketones were present at higher concentrations in the 1:1000 treatment combination than in the other combinations. Our findings emphasize the importance of selecting the appropriate proportions of L. delbrueckii subsp. bulgaricus and S. thermophilus for the starter culture in determining the final profile of volatiles and the overall flavor of dairy products.

  8. Mycobacterium avium subsp. paratuberculosis: presencia en los alimentos y su relación con la enfermedad de Crohn Mycobacterium avium subsp. paratuberculosis in food and its relationship with Crohn's disease

    Directory of Open Access Journals (Sweden)

    K. Cirone

    2007-03-01

    Full Text Available La paratuberculosis o enfermedad de Johne es una enteritis crónica producida por Mycobacterium avium subsp. paratuberculosis, que afecta a bovinos y a otras especies. En la Argentina se ha caracterizado en rodeos bovinos y de ciervos, con aislamientos tipificados en distintos patrones genéticos. M. avium subsp. paratuberculosis ha sido vinculado en humanos con una inflamación crónica del intestino, denominada enfermedad de Crohn. Existen evidencias clínicas y experimentales que relacionan a M. avium subsp. paratuberculosis con la enfermedad en el humano, mediante su detección por PCR y por cultivo a partir de biopsias de órganos, de leche materna y de sangre de pacientes afectados. La leche y sus subproductos serían posibles fuentes de infección y se ha sugerido que M. avium subsp. paratuberculosis resistiría las condiciones de pasteurización. Diversos trabajos de investigación demostraron que esta micobacteria podría estar presente en leches comercializadas en diversos países, como Reino Unido, Estados Unidos, República Checa, y también en la Argentina. La presencia de M. avium subsp. paratuberculosis en productos lácteos y agua de consumo ha sido relacionada con la resistencia del microorganismo tanto a los procesos de elaboración como a los factores climáticos adversos, lo que enfatiza el rol de los alimentos y del agua como vías de transmisión al humano. Las investigaciones en curso podrían ratificar el riesgo y las implicancias de la exposición del humano a M. avium subsp. paratuberculosis a través de los alimentos y del agua contaminados, para determinar la importancia de la paratuberculosis como enfermedad zoonótica.Paratuberculosis or Johne's disease is a chronic enteritis of the cattle and other small ruminant animals caused by Mycobacterium avium subsp. paratuberculosis. In Argentina, the strains were characterized in beef and dairy cattle and deer in different genetic patterns by molecular tools. M. avium

  9. Distribution of Bacillus thuringiensis subsp. israelensis in Soil of a Swiss Wetland Reserve after 22 Years of Mosquito Control▿†

    Science.gov (United States)

    Guidi, Valeria; Patocchi, Nicola; Lüthy, Peter; Tonolla, Mauro

    2011-01-01

    Recurrent treatments with Bacillus thuringiensis subsp. israelensis are required to control the floodwater mosquito Aedes vexans that breeds in large numbers in the wetlands of the Bolle di Magadino Reserve in Canton Ticino, Switzerland. Interventions have been carried out since 1988. In the present study, the spatial distribution of resting B. thuringiensis subsp. israelensis spores in the soil was measured. The B. thuringiensis subsp. israelensis concentration was determined in soil samples collected along six transects covering different elevations within the periodically flooded zones. A total of 258 samples were processed and analyzed by quantitative PCR that targeted an identical fragment of 159 bp for the B. thuringiensis subsp. israelensis cry4Aa and cry4Ba genes. B. thuringiensis subsp. israelensis spores were found to persist in soils of the wetland reserve at concentrations of up to 6.8 log per gram of soil. Continuous accumulation due to regular treatments could be excluded, as the decrease in spores amounted to 95.8% (95% confidence interval, 93.9 to 97.7%). The distribution of spores was correlated to the number of B. thuringiensis subsp. israelensis treatments, the elevation of the sampling point, and the duration of the flooding periods. The number of B. thuringiensis subsp. israelensis treatments was the major factor influencing the distribution of spores in the different topographic zones (P thuringiensis subsp. israelensis spores are rather immobile after their introduction into the environment. PMID:21498758

  10. Asplenium x sleepiae nothosubsp krameri (A-foreziense x A-obovatum subsp obovatum), a fern hybrid new for France (Aspleniaceae, Pteridophyta)

    NARCIS (Netherlands)

    Prelli, R; Rasbach, H; Viane, R

    1998-01-01

    A mixed population of several Asplenium taxa was studied near Roquebrune-sur-Argens (France, Var). A. foreziense, A. obovatum subsp. obovatum and A. obovatum subsp. lanceolatum were identified in the field and then confirmed by spore measurement. The presence of two hybrids within the population was

  11. Characterization of Monoclonal Antibodies Specific for Erwinia carotovora subsp. atroseptica and Comparison of Serological Methods for Its Sensitive Detection on Potato Tubers

    Science.gov (United States)

    Gorris, María Teresa; Alarcon, Benito; Lopez, María M.; Cambra, Mariano

    1994-01-01

    Seven monoclonal antibodies (MAbs) to Erwinia carotovora subsp. atroseptica have been produced. One, called 4G4, reacted with high specificity for serogroup I of E. carotovora subsp. atroseptica, the most common serogroup on potato tubers in different serological assays. Eighty-six strains belonging to different E. carotovora subsp. atroseptica serogroups were assayed. Some strains of serogroup XXII also reacted positively. No cross-reactions were observed against other species of plant pathogenic bacteria or 162 saprophytic bacteria from potato tubers. Only one strain of E. chrysanthemi from potato cross-reacted. A comparison of several serological techniques to detect E. carotovora subsp. atroseptica on potato tubers was performed with MAb 4G4 or polyclonal antibodies. The organism was extracted directly from potato peels of artificially inoculated tubers by soaking or selective enrichment under anaerobiosis in a medium with polypectate. MAb 4G4 was able to detect specifically 240 E. carotovora subsp. atroseptica cells per ml by indirect immunofluorescence and immunofluorescence colony staining and after soaking by ELISA-DAS (double-antibody sandwich enzyme-linked immunosorbent assay) after enrichment. The same amount of cells was detected by using immunolectrotransfer with polyclonal antibodies, and E. carotovora subsp. atroseptica and subsp. carotovora were distinguished by the latter technique. ELISA-DAS using MAb 4G4 with an enrichment step also efficiently detected E. carotovora subsp. atroseptica in naturally infected tubers and plants. PMID:16349293

  12. Detection of Mycobacterium ulcerans subsp. shinshuense DNA from a water channel in familial Buruli ulcer cases in Japan.

    Science.gov (United States)

    Luo, Yuqian; Degang, Yang; Ohtsuka, Mikio; Ishido, Yuko; Ishii, Norihisa; Suzuki, Koichi

    2015-01-01

    To determine if Mycobacterium ulcerans subsp. shinshuense is present in the common living environment in a case of familial concurrent occurrence of Buruli ulcer in Japan. Environmental samples were collected primarily from a water channel in the patient's residence. A combination of whole-genome amplification, touchdown PCR and DNA sequencing was used to detect M. ulcerans subsp. shinshuense DNA. M. ulcerans subsp. shinshuense DNA was detected in a crayfish sampled in July, but not in other samples including a crayfish sampled in October. These findings support an association between contaminated aquatic environments and Buruli ulcer in Japan. The data also suggest dynamic seasonal appearance of the pathogen in the environment may contribute to the seasonal variation of Buruli ulcer occurrence in Japan.

  13. H(+) -ATPase-defective variants of Lactobacillus delbrueckii subsp. bulgaricus contribute to inhibition of postacidification of yogurt during chilled storage.

    Science.gov (United States)

    Wang, Xinhui; Ren, Hongyang; Liu, Dayu; Wang, Bing; Zhu, Wenyou; Wang, Wei

    2013-02-01

    Continued acid production by Lactobacillus delbrueckii subsp. bulgaricus during the chilled storage of yogurt is the major cause of postacidification, resulting in a short shelf life. Two H(+) -ATPase defective variants of L. delbrueckii subsp. bulgaricus were successfully isolated and their H(+) -ATPase activities were reduced by 51.3% and 34.3%, respectively. It was shown that growth and acid production of variants were remarkably inhibited. The variants were more sensitive to acidic condition and had a significant rate for inactivation of H(+) -ATPase by N, N-dicyclohexylcarbodiimide (DCCD), along with a low H(+) -extrusion, suggesting that H(+) -ATPase is direct response for H(+) -extrusion. In addition, the variants were also more sensitive to NaCl, while H(+) -ATPase activities of variants and parent strain were significantly enhanced by NaCl stress. Obviously, H(+) -ATPase might be involved in Na(+) transportation. Furthermore, variants were inoculated in fermented milk to ferment yogurt. There was no significant difference in flavor, whereas the postacidification of yogurt during chilled storage was remarkably inhibited. It is suggested that application of L. delbrueckii subsp. bulgaricus with reduced H(+) -ATPase activity in yogurt fermentation is one of effect, economic and simple avenues of inhibiting postacidification of yogurt during refrigerated storage, giving a longer shelf life. During yogurt fermentation, continued acid production by Lactobacillus delbrueckii subsp. bulgaricus during the chilled storage of yogurt leads to milk fermentation with high postacidification, resulting in a short shelf life. In this work, 2 acid-sensitive variant strains of L. delbrueckii subsp. bulgaricus were isolated. The characteristics related to H(+) -ATPase were compared and it was observed that milk fermented by the variants had lower postacidification, giving a longer shelf life. Application of L. delbrueckii subsp. bulgaricus with reduced H(+) -ATPase activity

  14. Transcriptome-Based Characterization of Interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus in Lactose-Grown Chemostat Cocultures

    Science.gov (United States)

    Mendes, Filipa; Sieuwerts, Sander; de Hulster, Erik; Almering, Marinka J. H.; Luttik, Marijke A. H.; Pronk, Jack T.; Smid, Eddy J.; Bron, Peter A.

    2013-01-01

    Mixed populations of Saccharomyces cerevisiae yeasts and lactic acid bacteria occur in many dairy, food, and beverage fermentations, but knowledge about their interactions is incomplete. In the present study, interactions between Saccharomyces cerevisiae and Lactobacillus delbrueckii subsp. bulgaricus, two microorganisms that co-occur in kefir fermentations, were studied during anaerobic growth on lactose. By combining physiological and transcriptome analysis of the two strains in the cocultures, five mechanisms of interaction were identified. (i) Lb. delbrueckii subsp. bulgaricus hydrolyzes lactose, which cannot be metabolized by S. cerevisiae, to galactose and glucose. Subsequently, galactose, which cannot be metabolized by Lb. delbrueckii subsp. bulgaricus, is excreted and provides a carbon source for yeast. (ii) In pure cultures, Lb. delbrueckii subsp. bulgaricus grows only in the presence of increased CO2 concentrations. In anaerobic mixed cultures, the yeast provides this CO2 via alcoholic fermentation. (iii) Analysis of amino acid consumption from the defined medium indicated that S. cerevisiae supplied alanine to the bacterium. (iv) A mild but significant low-iron response in the yeast transcriptome, identified by DNA microarray analysis, was consistent with the chelation of iron by the lactate produced by Lb. delbrueckii subsp. bulgaricus. (v) Transcriptome analysis of Lb. delbrueckii subsp. bulgaricus in mixed cultures showed an overrepresentation of transcripts involved in lipid metabolism, suggesting either a competition of the two microorganisms for fatty acids or a response to the ethanol produced by S. cerevisiae. This study demonstrates that chemostat-based transcriptome analysis is a powerful tool to investigate microbial interactions in mixed populations. PMID:23872557

  15. Oral supplementation with Lactobacillus delbrueckii subsp. bulgaricus 8481 enhances systemic immunity in elderly subjects.

    Science.gov (United States)

    Moro-García, Marco Antonio; Alonso-Arias, Rebeca; Baltadjieva, Maria; Fernández Benítez, Carlos; Fernández Barrial, Manuel Amadeo; Díaz Ruisánchez, Enrique; Alonso Santos, Ricardo; Alvarez Sánchez, Magdalena; Saavedra Miján, Juan; López-Larrea, Carlos

    2013-08-01

    Throughout life, there is an aging of the immune system that causes impairment of its defense capability. Prevention or delay of this deterioration is considered crucial to maintain general health and increase longevity. We evaluated whether dietary supplementation with Lactobacillus delbrueckii subsp. bulgaricus 8481 could enhance the immune response in the elderly. This multi-center, double-blind, and placebo controlled study enrolled 61 elderly volunteers who were randomly assigned to receive either placebo or probiotics. Each capsule of probiotics contained at least 3 × 10(7)  L. delbrueckii subsp. bulgaricus 8481. Individuals in the study were administered three capsules per day for 6 months. Blood samples were obtained at baseline (time 0), end of month 3, and month 6. We characterized cell subpopulations, measured cytokines by flow cytometry, quantified T cell receptor excision circle (TREC) by real-time PCR (RT-PCR), and determined human β-defensin-2 (hBD-2) concentrations and human cytomegalovirus (CMV) titers by enzyme-linked immunosorbent assay (ELISA). Elderly responded to the intake of probiotic with an increase in the percentage of NK cells, an improvement in the parameters defining the immune risk profile (IRP), and an increase in the T cell subsets that are less differentiated. The probiotic group also showed decreased concentrations of the pro-inflammatory cytokine IL-8 but increased antimicrobial peptide hBD-2. These effects disappeared within 6 months of stopping the probiotic intake. Immunomodulation induced by L. delbrueckii subsp. bulgaricus 8481 could favor the maintenance of an adequate immune response, mainly by slowing the aging of the T cell subpopulations and increasing the number of immature T cells which are potential responders to new antigens.

  16. Inside the adaptation process of Lactobacillus delbrueckii subsp. lactis to bile.

    Science.gov (United States)

    Burns, Patricia; Sánchez, Borja; Vinderola, Gabriel; Ruas-Madiedo, Patricia; Ruiz, Lorena; Margolles, Abelardo; Reinheimer, Jorge; de los Reyes-Gavilán, Clara G

    2010-08-15

    Progressive adaptation to bile might render some lactobacilli able to withstand physiological bile salt concentrations. In this work, the adaptation to bile was evaluated on previously isolated dairy strains of Lactobacillus delbrueckii subsp. lactis 200 and L. delbrueckii subsp. lactis 200+, a strain derived thereof with stable bile-resistant phenotype. The adaptation to bile was obtained by comparing cytosolic proteomes of both strains grown in the presence or absence of bile. Proteomics were complemented with physiological studies on both strains focusing on glycolytic end-products, the ability to adhere to the human intestinal epithelial cell line HT29-MTX and survival to simulated gastrointestinal conditions. Protein pattern comparison of strains grown with and without bile allowed us to identify 9 different proteins whose production was regulated by bile in both strains, and 17 proteins that showed differences in their levels between the parental and the bile-resistant derivative. These included general stress response chaperones, proteins involved in transcription and translation, in peptidoglycan/exopolysaccharide biosynthesis, in the lipid and nucleotide metabolism and several glycolytic and pyruvate catabolism enzymes. Differences in the level of metabolic end-products of the sugar catabolism were found between the strains 200 and 200+. A decrease in the adhesion of both strains to the intestinal cell line was detected in the presence of bile. In simulated gastric and intestinal juices, a protective effect was exerted by milk improving the survival of both microorganisms. These results indicate that bile tolerance in L. delbrueckii subsp. lactis involves several mechanisms responding to the deleterious impact of bile salts on bacterial physiology. Copyright 2010 Elsevier B.V. All rights reserved.

  17. First report of olive leaf scorch in Brazil, associated with Xylella fastidiosa subsp. pauca

    Directory of Open Access Journals (Sweden)

    Helvécio Della COLETTA-FILHO

    2016-05-01

    Full Text Available Olea europaea (L. trees displaying leaf scorching symptoms, identical to those recently reported for olive trees colonized by Xylella fastidiosa in Southern Italy and also in Argentina, were observed in commercial orchards of two counties in Southeastern Brazil. PCR-based diagnosis using conserved primers for X. fastidiosa strains (RST31/33 and also specific to X. fastidiaosa subsp. pauca (CVC1/272-2 int were positive for all symptomatic tested samples (n = 8 of 9, but no template was obtained using twigs from asymptomatic trees (n = 20. Bacterial colonies were isolated from symptomatic tissues on culture medium and confirmed by PCR using the set of primers specific to X. fastidiosa subsp. pauca. Comparative sequence analyses of seven MLST loci amplified from one tripled passaged colony (MFG01 perfectly matched with sequences of alleles leuA #7, petC #6, malF#8, cysG#10, holC#11, nuoL#8, and gltT#8, the allelic profile of Sequence Type-ST16, which is represented by the strain COF0238 isolated from Coffea arabica (L. in Brazil (http://pubmlst.org/xfastidiosa/. Phylogenetic analysis placed the ST16 into subspecies pauca, but genetically closer to ST11 and ST13, both obtained from Citrus sinensis (L. trees with citrus variegated chlorosis. The results confirm the association of olive plants showing leaf scorching with the presence of X. fastidiosa subsp. pauca , and represent the first report of this bacterium in Brazilian olive orchards.

  18. Zoonotic necrotizing myositis caused by Streptococcus equi subsp. zooepidemicus in a farmer.

    Science.gov (United States)

    Kittang, Bård Reiakvam; Pettersen, Veronika Kuchařová; Oppegaard, Oddvar; Skutlaberg, Dag Harald; Dale, Håvard; Wiker, Harald G; Skrede, Steinar

    2017-02-15

    Streptococcus equi subsp. zooepidemicus is a beta-hemolytic group C streptococcus mainly causing infections in domesticated animals. Here we describe the first case of zoonotic necrotizing myositis caused by this bacterium. The patient was a 73-year-old, previously healthy farmer with two asymptomatic Shetland ponies in his stable. After close contact with the ponies while feeding them, he rapidly developed erythema of his left thigh and sepsis with multiple organ failure. The clinical course was severe and complicated, requiring repetitive surgical excision of necrotic muscle, treatment with vasopressors, mechanical ventilation and continuous venovenous hemofiltration, along with adjunctive hyperbaric oxygen therapy. The patient was discharged from hospital at day 30, without obvious sequelae. The streptococcal isolate was identified as Streptococcus equi by MALDI-ToF MS, and was later assigned subspecies identification as S. equi subsp. zooepidemicus. Multilocus sequence typing identified the strain as a novel sequence type (ST 364), closely related to types previously identified in horses and cattle. A focused proteomic analysis revealed that the ST 364 expressed putative virulence factors similar to that of Streptococcus pyogenes, including homologues of the M protein, streptodornases, interleukin 8-protease and proteins involved in the biosynthesis of streptolysin S. This case illustrates the zoonotic potential of S. equi subsp. zooepidemicus and the importance of early clinical recognition, rapid and radical surgical therapy, appropriate antibiotics and adequate supportive measures when necrotizing soft tissue infection is suspected. The expression of Streptococcus pyogenes-like putative virulence determinants in ST 364 might partially explain the fulminant clinical picture.

  19. Phylogeography and seed dispersal in islands: the case of Rumex bucephalophorus subsp. canariensis (Polygonaceae).

    Science.gov (United States)

    Talavera, María; Navarro-Sampedro, Laura; Ortiz, Pedro L; Arista, Montserrat

    2013-02-01

    Rumex bucephalophorus subsp. canariensis is an endemic taxon to Macaronesia with diaspore polymorphism. The origin and colonizing route of this taxon in Macaronesia was studied using molecular data and information on diaspore types. Amplified fragment length polymorphism (AFLP) was used in 260 plants from 22 populations of R. bucephalophorus subsp. canariensis, four from the Madeiran archipelago and 18 from the Canary archipelago. Diaspore production was analysed in 9-50 plants from each population used for AFLP analysis. One hundred and one plants from the Madeiran archipelago and 375 plants from the Canary Islands were studied. For each plant the type of diaspore produced was recorded. Overall populations had low genetic diversity but they showed a geographical pattern of genetic diversity that was higher in the older eastern islands than in the younger western ones. Two types of dispersible diaspores were found: in the eastern Canary islands (Lanzarote, Fuerteventura and Gran Canaria), plants produced exclusively long-dispersible diaspores, whereas in the western Canary islands (Tenerife, La Gomera, El Hierro) and the Madeiran archipelago plants produced exclusively short-dispersible diaspores. Genetically, the studied populations fell into four main island groups: Lanzarote-Fuerteventura, Gran Canaria, Tenerife-El Hierro and La Gomera-Madeira archipelago. A Moroccan origin of R. bucephalophorus subsp. canariensis is hypothesized with a colonization route from the eastern to the western islands. In addition, at least one gene flow event from La Gomera to the Madeiran archipelago has taken place. During the colonization process the type of dispersible diaspore changed so that dispersability decreased in populations of the westernmost islands.

  20. Characterization of a new bacteriocin, Carocin D, from Pectobacterium carotovorum subsp. carotovorum Pcc21.

    Science.gov (United States)

    Roh, Eunjung; Park, Tae-Ho; Kim, Myung-Il; Lee, Seungdon; Ryu, Sangryeol; Oh, Chang-Sik; Rhee, Sangkee; Kim, Doo-Ho; Park, Beom-Seok; Heu, Sunggi

    2010-11-01

    Two different bacteriocins, carotovoricin and carocin S1, had been found in Pectobacterium carotovorum subsp. carotovorum, which causes soft-rot disease in diverse plants. Previously, we reported that the particular strain Pcc21, producing only one high-molecular-weight bacteriocin, carried a new antibacterial activity against the indicator strain Pcc3. Here, we report that this new antibacterial activity is due to a new bacteriocin produced by strain Pcc21 and named carocin D. Carocin D is encoded by the caroDK gene located in the genomic DNA together with the caroDI gene, which seems to encode an immunity protein. N-terminal amino acid sequences of purified carocin D were determined by Edman degradation. In comparison with the primary translation product of caroDK, it was found that 8 amino acids are missing at the N terminus. This finding proved that carocin D is synthesized as a precursor peptide and that 8 amino acids are removed from its N terminus during maturation. Carocin D has two putative translocation domains; the N-terminal and C-terminal domains are homologous to those of Escherichia coli colicin E3 and Pseudomonas aeruginosa S-type pyocin, respectively. When caroDK and caroDI genes were transformed into carocin D-sensitive bacteria such as Pcc3, the bacteria became resistant to this bacteriocin. Carocin D has one putative DNase domain at the extreme C terminus and showed DNase activity in vitro. This bacteriocin had slight tolerance to heat but not to proteases. The caroDK gene was present in only 5 of 54 strains of P. carotovorum subsp. carotovorum. These results indicate that carocin D is a third bacteriocin found in P. carotovorum subsp. carotovorum, and this bacteriocin can be readily expressed in carocin D-sensitive nonpathogenic bacteria, which may have high potential as a biological control agent in the field.

  1. Characterization of a New Bacteriocin, Carocin D, from Pectobacterium carotovorum subsp. carotovorum Pcc21▿ †

    Science.gov (United States)

    Roh, Eunjung; Park, Tae-Ho; Kim, Myung-il; Lee, Seungdon; Ryu, Sangryeol; Oh, Chang-Sik; Rhee, Sangkee; Kim, Doo-Ho; Park, Beom-Seok; Heu, Sunggi

    2010-01-01

    Two different bacteriocins, carotovoricin and carocin S1, had been found in Pectobacterium carotovorum subsp. carotovorum, which causes soft-rot disease in diverse plants. Previously, we reported that the particular strain Pcc21, producing only one high-molecular-weight bacteriocin, carried a new antibacterial activity against the indicator strain Pcc3. Here, we report that this new antibacterial activity is due to a new bacteriocin produced by strain Pcc21 and named carocin D. Carocin D is encoded by the caroDK gene located in the genomic DNA together with the caroDI gene, which seems to encode an immunity protein. N-terminal amino acid sequences of purified carocin D were determined by Edman degradation. In comparison with the primary translation product of caroDK, it was found that 8 amino acids are missing at the N terminus. This finding proved that carocin D is synthesized as a precursor peptide and that 8 amino acids are removed from its N terminus during maturation. Carocin D has two putative translocation domains; the N-terminal and C-terminal domains are homologous to those of Escherichia coli colicin E3 and Pseudomonas aeruginosa S-type pyocin, respectively. When caroDK and caroDI genes were transformed into carocin D-sensitive bacteria such as Pcc3, the bacteria became resistant to this bacteriocin. Carocin D has one putative DNase domain at the extreme C terminus and showed DNase activity in vitro. This bacteriocin had slight tolerance to heat but not to proteases. The caroDK gene was present in only 5 of 54 strains of P. carotovorum subsp. carotovorum. These results indicate that carocin D is a third bacteriocin found in P. carotovorum subsp. carotovorum, and this bacteriocin can be readily expressed in carocin D-sensitive nonpathogenic bacteria, which may have high potential as a biological control agent in the field. PMID:20870796

  2. Microencapsulation of Bifidobacterium animalis subsp. lactis and Lactobacillus acidophilus in cocoa butter using spray chilling technology

    Science.gov (United States)

    Pedroso, D.L.; Dogenski, M.; Thomazini, M.; Heinemann, R.J.B.; Favaro-Trindade, C.S.

    2013-01-01

    In the present study, the cells of Bifidobacterium animalis subsp. lactis (BI-01) and Lactobacillus acidophilus (LAC-04) were encapsulated in cocoa butter using spray-chilling technology. Survival assays were conducted to evaluate the resistance of the probiotics to the spray-chilling process, their resistance to the simulated gastric and intestinal fluids (SGF and SIF), and their stability during 90 days of storage. The viability of the cells was not affected by microencapsulation. The free and encapsulated cells of B. animalis subsp. lactis were resistant to both SGF and SIF. The micro-encapsulated cells of L. acidophilus were more resistant to SGF and SIF than the free cells; the viability of the encapsulated cells was enhanced by 67%, while the free cells reached the detection limit of the method (103 CFU/g). The encapsulated probiotics were unstable when they were stored at 20 °C. The population of encapsulated L. acidophilus decreased drastically when they were stored at 7 °C; only 20% of cells were viable after 90 days of storage. The percentage of viable cells of the encapsulated B. animalis subsp.lactis, however, was 72% after the same period of storage. Promising results were obtained when the microparticles were stored at −18 °C; the freeze granted 90 days of shelf life to the encapsulated cells. These results suggest that the spray-chilling process using cocoa butter as carrier protects L. acidophilus from gastrointestinal fluids. However, the viability of the cells during storage must be improved. PMID:24516445

  3. Microencapsulation of Bifidobacterium animalis subsp. lactis and Lactobacillus acidophilus in cocoa butter using spray chilling technology

    Directory of Open Access Journals (Sweden)

    D.L. Pedroso

    2013-09-01

    Full Text Available In the present study, the cells of Bifidobacterium animalis subsp. lactis (BI-01 and Lactobacillus acidophilus (LAC-04 were encapsulated in cocoa butter using spray-chilling technology. Survival assays were conducted to evaluate the resistance of the probiotics to the spray-chilling process, their resistance to the simulated gastric and intestinal fluids (SGF and SIF, and their stability during 90 days of storage. The viability of the cells was not affected by microencapsulation. The free and encapsulated cells of B. animalis subsp. lactis were resistant to both SGF and SIF. The micro-encapsulated cells of L. acidophilus were more resistant to SGF and SIF than the free cells; the viability of the encapsulated cells was enhanced by 67%, while the free cells reached the detection limit of the method (10³ CFU/g. The encapsulated probiotics were unstable when they were stored at 20 °C. The population of encapsulated L. acidophilus decreased drastically when they were stored at 7 °C; only 20% of cells were viable after 90 days of storage. The percentage of viable cells of the encapsulated B. animalis subsp.lactis, however, was 72% after the same period of storage. Promising results were obtained when the microparticles were stored at -18 °C; the freeze granted 90 days of shelf life to the encapsulated cells. These results suggest that the spray-chilling process using cocoa butter as carrier protects L. acidophilus from gastrointestinal fluids. However, the viability of the cells during storage must be improved.

  4. Exopolysaccharide and extracellular metabolite production by Lactobacillus delbrueckii subsp. bulgaricus, grown on lactose in continuous culture.

    Science.gov (United States)

    Welman, Alan; Maddox, Ian; Archer, Richard

    2003-09-01

    Lactobacillus delbrueckii subsp. bulgaricus NCFB 2483, when grown on lactose in continuous culture, showed increasing specific yields and volumetric productivities of exopolysaccharide (EPS) with increasing dilution rate. Specific and volumetric productivities of lactate and galactose, as extracellular metabolites, increased in response to the incremental changes in the dilution rate up to 0.4 h(-1). Elevated Y(p/s) values determined for EPS (0.025 g EPS x g lactose(-1)) at the dilution rates of 0.3 h(-1)-0.4 h(-1), relative to those determined at lower dilution rates, suggest a diversion of carbon flux towards EPS being associated with the higher rates of growth.

  5. DEUTEROCOHNIA MEZIANA SUBSP. VALLEGRANDENSIS (BROMELIACEAE, NUEVA SUBESPECIE Y PRIMER REGISTRO DE LA ESPECIE PARA ARGENTINA

    Directory of Open Access Journals (Sweden)

    Silvia E. Gómez Romero

    2015-01-01

    Full Text Available Se describe e ilustra una nueva subespecie de Jujuy (Argentina, Deuterocohnia meziana subsp. vallegrandensis, y se presenta un mapa con su distribución geográfica. La misma se distingue claramente del resto de las subespecies por sus sépalos castaño purpúreos y pétalos del mismo color o rojizos con ápice verdoso. Se cita por primera vez esta especie para la Flora Argentina. Se incluye una clave para Deuterocohnia en Argentina y una tabla con los caracteres que distinguen las cinco subespecies de D. meziana. Se designa un lectotipo para Deuterocohnia meziana.

  6. Desenvolvimento de vacina recombinante de proteína M de Streptococcus equi subsp. equi

    OpenAIRE

    MACIEL, Liana Flores

    2012-01-01

    A equinocultura no Brasil ganha espaço em setores ligados ao lazer, cultura e turismo, sendo responsável por milhões de empregos. A Adenite Equina causada pelo Streptococcus equi subsp. equi é uma doença do aparelho respiratório de elevado impacto econômico, gerando gastos com mão-de-obra e perda de desempenho dos animais. Para amenizar este problema, medidas profiláticas são importantes, como por exemplo, a vacinação. Porém, as vacinas disponíveis no mercado protegem apenas 50% dos animai...

  7. First report of Cowpea mild mottle Carlavirus on yardlong bean (Vigna unguiculata subsp. sesquipedalis) in Venezuela.

    Science.gov (United States)

    Brito, Miriam; Fernández-Rodríguez, Thaly; Garrido, Mario José; Mejías, Alexander; Romano, Mirtha; Marys, Edgloris

    2012-12-14

    Yardlong bean (Vigna unguiculata subsp. sesquipedalis) plants with virus-like systemic mottling and leaf distortion were observed in both experimental and commercial fields in Aragua State, Venezuela. Symptomatic leaves were shown to contain carlavirus-like particles. RT-PCR analysis with carlavirus-specific primers was positive in all tested samples. Nucleotide sequences of the obtained amplicons showed 84%-74% similarity to corresponding sequences of Cowpea mild mottle virus (CPMMV) isolates deposited in the GenBank database. This is the first report of CPMMV in Venezuela and is thought to be the first report of CPMMV infecting yardlong bean.

  8. First Report of Cowpea Mild Mottle Carlavirus on Yardlong Bean (Vigna unguiculata subsp. sesquipedalis in Venezuela

    Directory of Open Access Journals (Sweden)

    Edgloris Marys

    2012-12-01

    Full Text Available Yardlong bean (Vigna unguiculata subsp. sesquipedalis plants with virus-like systemic mottling and leaf distortion were observed in both experimental and commercial fields in Aragua State, Venezuela. Symptomatic leaves were shown to contain carlavirus-like particles. RT-PCR analysis with carlavirus-specific primers was positive in all tested samples. Nucleotide sequences of the obtained amplicons showed 84%–74% similarity to corresponding sequences of Cowpea mild mottle virus (CPMMV isolates deposited in the GenBank database. This is the first report of CPMMV in Venezuela and is thought to be the first report of CPMMV infecting yardlong bean.

  9. [First case report of Streptococcus equi subsp. zooepidemicus post-infectious acute glomerulonephritis in France].

    Science.gov (United States)

    Le-Berre, Nicolas; Filipozzi, Pierre; Martin, Laurent; Frimat, Luc; Girerd, Sophie

    2017-02-01

    Post-infectious glomerulonephritis has become exceptional in France because streptococcus infections are well-treated. When they appear, clinical and biological symptoms are mostly typical and associate acute nephritic syndrome, acute renal failure, proteinuria, hematuria and low complement. We are reporting the first French case of acute post-infectious glomerulonephritis related to Streptococcus equi subsp. zooepidemicus, which is commonly found in horses and rarely in human pathology, and of which contamination is by direct contact with sick horses or by ingestion of non-pasteurized milk. Copyright © 2016 Association Société de néphrologie. Published by Elsevier SAS. All rights reserved.

  10. Genetic diversity of Streptococcus equi subsp. zooepidemicus and doxycycline resistance in kennelled dogs.

    Science.gov (United States)

    Chalker, Victoria J; Waller, Andrew; Webb, Katy; Spearing, Emma; Crosse, Patricia; Brownlie, Joe; Erles, Kerstin

    2012-06-01

    The genetic diversity and antibiotic resistance profiles of 38 Streptococcus equi subsp. zooepidemicus isolates were determined from a kennelled canine population during two outbreaks of hemorrhagic pneumonia (1999 to 2002 and 2007 to 2010). Analysis of the szp gene hypervariable region and the 16S-23S rRNA intergenic spacer region and multilocus sequence typing (MLST) indicated a predominant tetO-positive, doxycycline-resistant ST-10 strain during 1999 to 2002 and a predominant tetM-positive doxycycline-resistant ST-62 strain during 2007 to 2010.

  11. Homology Modeling and Functional Characterization of PR-1a Protein of Hordeum vulgare subsp. Vulgare

    OpenAIRE

    Aslanzadeh, Vahid; Ghaderian, Mostafa

    2012-01-01

    Pathogenesis-related protein 1a of Hordeum vulgare subsp. Vulgare (HvPR-1a) is induced by various pathogens and stress related factors. It plays important roles in plant defense system. Since the discovery of HvPR-1a a great deal of research has been focused on its isolation and characterization. However, three dimensional structure of HvPR-1a is still unknown. 3D structure can be used for determining protein function, and identifying novel protein folds and potential targets for regulation. ...

  12. Association between milk antibody and interferon-gamma responses in cattle from Mycobacterium avium subsp. paratuberculosis infected herds

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Jungersen, Gregers; Nielsen, Søren Saxmose

    2009-01-01

    Paratuberculosis is a chronic infection of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). It is possible to detect infection with paratuberculosis at different stages of disease by means of various diagnostic test strategies. The objective of the present study was to evalu......Paratuberculosis is a chronic infection of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). It is possible to detect infection with paratuberculosis at different stages of disease by means of various diagnostic test strategies. The objective of the present study...

  13. Isolation of Streptococcus equi subsp. equi from thoroughbred horses in a racehorse-breeding area of Japan.

    Science.gov (United States)

    Anzai, T; Nakanishi, A; Wada, R; Higuchi, T; Hagiwara, S; Takazawa, M; Oobayashi, K; Inoue, T

    1997-11-01

    For determination whether strangles has invaded the Hidaka district of Hokkaido, the main racehorse-breeding area of Japan, a epizootiological survey with bacterial isolation was carried out during the breeding season in 1995. Streptococcus equi subsp. equi, which is the causative agent of strangles, was isolated from two Thoroughbred horses with submandibular lymphadenitis. Isolates were identified by serological grouping, biochemical tests and analysis of cell surface proteins by Western immunoblotting. Through this survey, it revealed that S. equi subsp. equi has invaded the Hidaka district and that strangles has become prevalent in racehorse-breeding farms in this area.

  14. Proteomes of Lactobacillus delbrueckii subsp. bulgaricus LBB.B5 Incubated in Milk at Optimal and Low Temperatures.

    Science.gov (United States)

    Yin, Xiaochen; Salemi, Michelle R; Phinney, Brett S; Gotcheva, Velitchka; Angelov, Angel; Marco, Maria L

    2017-01-01

    We identified the proteins synthesized by Lactobacillus delbrueckii subsp. bulgaricus strain LBB.B5 in laboratory culture medium (MRS) at 37°C and milk at 37 and 4°C. Cell-associated proteins were measured by gel-free, shotgun proteomics using high-performance liquid chromatography coupled with tandem mass spectrophotometry. A total of 635 proteins were recovered from all cultures, among which 72 proteins were milk associated (unique or significantly more abundant in milk). LBB.B5 responded to milk by increasing the production of proteins required for purine biosynthesis, carbohydrate metabolism (LacZ and ManM), energy metabolism (TpiA, PgK, Eno, SdhA, and GapN), amino acid synthesis (MetE, CysK, LBU0412, and AspC) and transport (GlnM and GlnP), and stress response (Trx, MsrA, MecA, and SmpB). The requirement for purines was confirmed by the significantly improved cell yields of L. delbrueckii subsp. bulgaricus when incubated in milk supplemented with adenine and guanine. The L. delbrueckii subsp. bulgaricus -expressed proteome in milk changed upon incubation at 4°C for 5 days and included increased levels of 17 proteins, several of which confer functions in stress tolerance (AddB, UvrC, RecA, and DnaJ). However, even with the activation of stress responses in either milk or MRS, L. delbrueckii subsp. bulgaricus did not survive passage through the murine digestive tract. These findings inform efforts to understand how L. delbrueckii subsp. bulgaricus is adapted to the dairy environment and its implications for its health-benefiting properties in the human digestive tract. IMPORTANCE Lactobacillus delbrueckii subsp. bulgaricus has a long history of use in yogurt production. Although commonly cocultured with Streptococcus salivarius subsp. thermophilus in milk, fundamental knowledge of the adaptive responses of L. delbrueckii subsp. bulgaricus to the dairy environment and the consequences of those responses on the use of L. delbrueckii subsp. bulgaricus as

  15. Salmonella enterica suppresses Pectobacterium carotovorum subsp. carotovorum population and soft rot progression by acidifying the microaerophilic environment.

    Science.gov (United States)

    Kwan, Grace; Charkowski, Amy O; Barak, Jeri D

    2013-02-12

    Although enteric human pathogens are usually studied in the context of their animal hosts, a significant portion of their life cycle occurs on plants. Plant disease alters the phyllosphere, leading to enhanced growth of human pathogens; however, the impact of human pathogens on phytopathogen biology and plant health is largely unknown. To characterize the interaction between human pathogens and phytobacterial pathogens in the phyllosphere, we examined the interactions between Pectobacterium carotovorum subsp. carotovorum and Salmonella enterica or Escherichia coli O157:H7 with regard to bacterial populations, soft rot progression, and changes in local pH. The presence of P. carotovorum subsp. carotovorum enhanced the growth of both S. enterica and E. coli O157:H7 on leaves. However, in a microaerophilic environment, S. enterica reduced P. carotovorum subsp. carotovorum populations and soft rot progression by moderating local environmental pH. Reduced soft rot was not due to S. enterica proteolytic activity. Limitations on P. carotovorum subsp. carotovorum growth, disease progression, and pH elevation were not observed on leaves coinoculated with E. coli O157:H7 or when leaves were coinoculated with S. enterica in an aerobic environment. S. enterica also severely undermined the relationship between the phytobacterial population and disease progression of a P. carotovorum subsp. carotovorum budB mutant defective in the 2,3-butanediol pathway for acid neutralization. Our results show that S. enterica and E. coli O157:H7 interact differently with the enteric phytobacterial pathogen P. carotovorum subsp. carotovorum. S. enterica inhibition of soft rot progression may conceal a rapidly growing human pathogen population. Whereas soft rotted produce can alert consumers to the possibility of food-borne pathogens, healthy-looking produce may entice consumption of contaminated vegetables. Salmonella enterica and Escherichia coli O157:H7 may use plants to move between animal

  16. Extracellular Nucleases of Streptococcus equi subsp. zooepidemicus Degrade Neutrophil Extracellular Traps and Impair Macrophage Activity of the Host.

    Science.gov (United States)

    Ma, Fang; Guo, Xiao; Fan, Hongjie

    2017-01-15

    The pathogen Streptococcus equi subsp. zooepidemicus is associated with a wide range of animals, including humans, and outbreaks frequently occur in pigs, equines, and goats. Thus far, few studies have assessed interactions between the host immune system and S. equi subsp. zooepidemicus and how these interactions explain the wide host spectrum of S. equi subsp. zooepidemicus Neutrophils, the first line of innate immunity, possess a defense mechanism called neutrophil extracellular traps (NETs), which primarily consist of DNA and granule proteins that trap bacteria via charge interactions. Extracellular nucleases play important roles in the degradation of the DNA backbone of NETs. Here, two related extracellular nucleases, nuclease and 5'-nucleotidase (named ENuc and 5Nuc, respectively, in this study), were identified as being encoded by the SESEC_RS04165 gene and the SESEC_RS05720 gene (named ENuc and 5Nuc, respectively), and three related gene deletion mutant strains, specifically, the single-mutant ΔENuc and Δ5Nuc strains and the double-mutant ΔENuc Δ5Nuc strain, were constructed. The ΔENuc and Δ5Nuc single-mutant strains and the ΔENuc Δ5Nuc double-mutant strain demonstrated lower virulence than wild-type S. equi subsp. zooepidemicus when the mouse survival rate was evaluated postinfection. Furthermore, wild-type S. equi subsp. zooepidemicus more frequently traversed the bloodstream and transferred to other organs. Wild-type S. equi subsp. zooepidemicus induced fewer NETs and was able to survive in NETs, whereas only 40% of the ΔENuc Δ5Nuc double-mutant cells survived. S. equi subsp. zooepidemicus degraded the NET DNA backbone and produced deoxyadenosine, primarily through the action of ENuc and/or 5Nuc. However, the double-mutant ΔENuc Δ5Nuc strain lost the ability to degrade NETs into deoxyadenosine. Deoxyadenosine decreased RAW 264.7 cell phagocytosis to 40% of that of normal macrophages. Streptococcus equi subsp. zooepidemicus causes serious

  17. Development of a pentaplex PCR assay for the simultaneous detection of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. helveticus, L. fermentum in whey starter for Grana Padano cheese.

    Science.gov (United States)

    Cremonesi, Paola; Vanoni, Laura; Morandi, Stefano; Silvetti, Tiziana; Castiglioni, Bianca; Brasca, Milena

    2011-03-30

    A pentaplex PCR assay for the rapid, selective and simultaneous detection of Lactobacillus helveticus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and L. fermentum, was developed. The target sequences were a group of genes coding for beta-galactosidase production (S. thermophilus and L. delbrueckii subsp. bulgaricus), for cell-enveloped associated proteinase synthesis (L. helveticus), for dipeptide transport system production (L. delbrueckii subsp. lactis) and for arginine-ornithine antiporter protein production (L. fermentum). The analytical specificity of the assay was evaluated with 5 reference strains and 140 lactic acid bacterial strains derived from raw milk cheeses and belonging to the Lactobacillus, Streptococcus, Lactococcus and Enterococcus genera. The identification limit for each target strain was 10(3)CFU/ml. This new molecular assay was used to investigate the LAB population by direct extraction of DNA from the 12 whey cultures for Grana Padano. The pentaplex PCR assay revealed a good correspondence with microbiological analyses and allowed to identify even minor LAB community members which, can be out-competed in vitro by numerically more abundant microbial species. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Membrane protein profiling of Acidovorax avenae subsp. avenae under various growth conditions.

    Science.gov (United States)

    Li, Bin; Wang, Li; Ibrahim, Muhammad; Ge, Mengyu; Wang, Yanli; Mannan, Shazia; Asif, Muhammad; Sun, Guochang

    2015-06-01

    Membrane proteins (MPs) of plant pathogenic bacteria have been reported to be able to regulate many essential cellular processes associated with plant disease. The aim of the current study was to examine and compare the expression of MPs of the rice bacterial pathogen Acidovorax avenae subsp. avenae strain RS-1 under Luria-Bertani (LB) medium, M9 medium, in vivo rice plant conditions and leaf extract (LE) medium mimicking in vivo plant condition. Proteomic analysis identified 95, 72, 75, and 87 MPs under LB, in vivo, M9 and LE conditions, respectively. Among them, six proteins were shared under all tested growth conditions designated as abundant class of proteins. Twenty-six and 21 proteins were expressed uniquely under in vivo versus LB medium and LE versus M9 medium, respectively, with 17 proteins common among these uniquely induced proteins. Moreover, most of the shared proteins are mainly related to energy metabolism, transport of small molecules, protein synthesis and secretion as well as virulence such as NADH, OmpA, secretion proteins. Therefore, the result of this study not only suggests that it may be an alternate method to analyze the in vivo expression of proteins by using LE medium to mimic plant conditions, but also reveals that the two sets of differentially expressed MPs, in particular the common MPs between them, might be important in energy metabolism, stress response and virulence of A. avenae subsp. avenae strain RS-1.

  19. Application of Raman spectroscopy for direct analysis of Carlina acanthifolia subsp. utzka root essential oil.

    Science.gov (United States)

    Strzemski, Maciej; Wójciak-Kosior, Magdalena; Sowa, Ireneusz; Agacka-Mołdoch, Monika; Drączkowski, Piotr; Matosiuk, Dariusz; Kurach, Łukasz; Kocjan, Ryszard; Dresler, Sławomir

    2017-11-01

    Carlina genus plants e.g. Carlina acanthifolia subsp. utzka have been still used in folk medicine of many European countries and its biological activity is mostly associated with root essential oils. In the present paper, Raman spectroscopy (RS) was applied for the first time for evaluation of essential oil distribution in root of C. acnthifolia subsp. utzka and identification of root structures containing the essential oil. Furthermore, RS technique was applied to assess chemical stability of oil during drying of plant material or distillation process. Gas chromatography-mass spectrometry was used for qualitative and quantitative analysis of the essential oil. The identity of compounds was confirmed using Raman, ATR-IR and NMR spectroscopy. Carlina oxide was found to be the main component of the oil (98.96% ± 0.15). The spectroscopic study showed the high stability of essential oil and Raman distribution analysis indicated that the oil reservoirs were localized mostly in the structures of outer layer of the root while the inner part showed nearly no signal assigned to the oil. Raman spectroscopy technique enabled rapid, non-destructive direct analysis of plant material with minimal sample preparation and allowed straightforward, unambiguous identification of the essential oil in the sample. Copyright © 2017. Published by Elsevier B.V.

  20. Bejel in Cuba: molecular identification of Treponema pallidum subsp. endemicum in patients diagnosed with venereal syphilis.

    Science.gov (United States)

    Noda, A A; Grillová, L; Lienhard, R; Blanco, O; Rodríguez, I; Šmajs, D

    2018-02-16

    Bejel, caused by Treponema pallidum subsp. endemicum (TEN), was until now considered as a non-venereal disease endemic in areas with hot and dry climates. This study has identified TEN in clinical samples from Cuban patients previously diagnosed with syphilis. We performed sequencing-based molecular typing on 92 samples from Cuban individuals diagnosed with syphilis. Moreover, to differentiate T. pallidum subspecies, multi-locus sequence analysis (MLSA) was designed and was applied to suspicious samples. Nine samples, from six patients, had a nucleotide sequence similarity (at all typing loci) to the Bosnia A genome, which is the infectious agent of bejel. Additionally, MLSA clearly supported a TEN classification for the treponemal samples. Clinical and epidemiological data from the six patients also suggested sexual transmission of bejel as well as the endemicity of this rare treponematosis in Cuba. Molecular identification of Treponema pallidum subsp. endemicum, the agent of bejel, in Cuban patients diagnosed with syphilis indicates the clear limitations of a diagnosis based exclusively on serology, geographical occurrence, clinical symptoms and anamnestic data. This finding has important implications for Global Public Health Systems, including paradigm changes regarding the location of endemic outbreaks, clinical aspects and transmission of this neglected disease. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  1. Three new flavonoids from the seeds of Hippophae rhamnoides subsp. sinensis.

    Science.gov (United States)

    Zhang, Jing; Gao, Wen; Cao, Min-Sheng; Kong, De-Yun

    2012-01-01

    To study the chemical constituents of the seeds of Hippophae rhamnoides subsp. sinensis, three new flavonoids acylated with one monoterpenic acid, named 3-O-β-D-glucosyl-kaempferol-7-O-{2-O-[2(E)-2,6-dimethyl-6-hydroxy-2,7-octadienoyl]}-α-L-rhamnoside (3), 3-O-β-D-sophorosyl-kaempferol-7-O-{3-O-[2(E)-2,6-dimethyl-6-hydroxy-2,7-octadienoyl]}-α-L-rhamnoside (4), and 3-O-β-D-sophorosyl-kaempferol-7-O-{2-O-[2(E)-2,6-dimethyl-6-hydroxy-2,7-octadienoyl]}-α-L-rhamnoside (5), together with four known compounds, were isolated from the seeds of H. rhamnoides subsp. sinensis. Compounds 1 and 2 are reported for the first time from this genus. Their structures were elucidated on the basis of chemical and spectral analysis, including 1D and 2D NMR and HR-MS, and by comparison with literature data.

  2. Optimization of Exopolysaccharide Production by Lactobacillus delbrueckii subsp. bulgaricus RR Grown in a Semidefined Medium

    Science.gov (United States)

    Kimmel, Stacy A.; Roberts, Robert F.; Ziegler, Gregory R.

    1998-01-01

    The optimal fermentation temperature, pH, and Bacto-casitone (Difco Laboratories, Detroit, Mich.) concentration for production of exopolysaccharide by Lactobacillus delbrueckii subsp. bulgaricus RR in a semidefined medium were determined by using response surface methods. The design consisted of 20 experiments, 15 unique combinations, and five replications. All fermentations were conducted in a fermentor with a 2.5-liter working volume and were terminated when 90% of the glucose in the medium had been consumed. The population of L. delbrueckii subsp. bulgaricus RR and exopolysaccharide content were measured at the end of each fermentation. The optimum temperature, pH, and Bacto-casitone concentration for exopolysaccharide production were 38°C, 5, and 30 g/liter, respectively, with a predicted yield of 295 mg of exopolysaccharide/liter. The actual yield under these conditions was 354 mg of exopolysaccharide/liter, which was within the 95% confidence interval (217 to 374 mg of exopolysaccharide/liter). An additional experiment conducted under optimum conditions showed that exopolysaccharide production was growth associated, with a specific production at the endpoint of 101.4 mg/g of dry cells. Finally, to obtain material for further characterization, a 100-liter fermentation was conducted under optimum conditions. Twenty-nine grams of exopolysaccharide was isolated from centrifuged, ultrafiltered fermentation broth by ethanol precipitation. PMID:9464404

  3. Production of lactic acid from whey using Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus

    Directory of Open Access Journals (Sweden)

    Adriana M. Rojas

    2015-09-01

    Full Text Available The main objective of this research was to determine the proper growth conditions of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus for the production of lactic acid using serum as substract. This serum was obtain from the department of Cesar, Colombia. Lactic acid is the result of the extraction and purification of fermentation broths in which bacteria Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus are used, which are usually used for the production of yogurt. The substrate was supplemented with yeast extract, ammonium phosphate as a nitrogen source, and calcium carbonate as a neutralizer, in order to optimize the consumption, by the bacteria, of the main carbohydrate present in serum (lactose. During the fermentation (up to 72 h the inoculums concentration, and temperature were controlled. Purification consisted in esterification, filtration of solids formed during the reaction, and removing of water by evaporation and nitrogen influx. Finally, lactic acid was obtained with 78,0% purity (36.7 g/L, which was characterized by infrared spectroscopy

  4. In vitro multiple shoot regeneration from hypocotyl in Menta spicata subsp. spicata

    OpenAIRE

    Özdemir, Fethi; Yıldırım, Mehmet; Pourali Kahriz, Mahsa

    2015-01-01

    Mentha spicata subsp. spicata tıbbi ve aromatik bir bitkidir. Bu çalışmada Mentha spicata subsp. spicata’nın in vitro’ da çoğaltımı amacıyla hipokotil eksplantları BAP ve NAA ’nın farklı kombinasyonlarını içeren, %3 sukroz ilave edilmiş ve % 0.65 agar ile katılaştırılmış MS besin ortamında kültüre alınmıştır. Araştırma sonuçları kallus oluşum yüzdesi, rejenere sürgün yüzdesi, eksplant başına düşen ortalama sürgün sayısı, ortalama sürgün uzunluğu, eksplant başına düşen ortalama kök sayısı ve o...

  5. The Cry Toxin Operon of Clostridium bifermentans subsp. malaysia Is Highly Toxic to Aedes Larval Mosquitoes

    Science.gov (United States)

    Qureshi, Nadia; Chawla, Swati; Likitvivatanavong, Supaporn; Lee, Han Lim

    2014-01-01

    The management and control of mosquito vectors of human disease currently rely primarily on chemical insecticides. However, larvicidal treatments can be effective, and if based on biological insecticides, they can also ameliorate the risk posed to human health by chemical insecticides. The aerobic bacteria Bacillus thuringiensis and Lysinibacillus sphaericus have been used for vector control for a number of decades. But a more cost-effective use would be an anaerobic bacterium because of the ease with which these can be cultured. More recently, the anaerobic bacterium Clostridium bifermentans subsp. malaysia has been reported to have high mosquitocidal activity, and a number of proteins were identified as potentially mosquitocidal. However, the cloned proteins showed no mosquitocidal activity. We show here that four toxins encoded by the Cry operon, Cry16A, Cry17A, Cbm17.1, and Cbm17.2, are all required for toxicity, and these toxins collectively show remarkable selectivity for Aedes rather than Anopheles mosquitoes, even though C. bifermentans subsp. malaysia is more toxic to Anopheles. Hence, toxins that target Anopheles are different from those expressed by the Cry operon. PMID:25002432

  6. Desulfovibrio oceani subsp. oceani sp. nov., subsp. nov and Desulfovibrio oceani subsp. galateae subsp. nov., novel sulfate-reducing bacteria isolated from the oxygen minimum zone off the coast of Peru

    DEFF Research Database (Denmark)

    Finster, Kai; Kjeldsen, Kasper Urup

    2010-01-01

    at 20°C at pH 7.0-8.0 and at 2.5-3.5% NaCl (w/v). The strains grew by utilizing hydrogen/acetate, C3-4 fatty acids, amino acids and glycerol as electron acceptors for sulfate reduction. Fumarate, lactate and pyruvate supported fermentative growth. Sulfate, sulfite, thiosulfate and taurin supported......%) for strain I.9.1T. The G+C contents of their genomic DNA were 45-46 mol%. Phylogenetic analysis of 16S rRNA and dsrAB gene sequences showed that both strains belong to the genus Desulfovibrio. Desulfovibrio acrylicus DSM 10141T and Desulfovibrio marinisediminis JCM 14577T represented their closest validly.......1T represent a novel species for which the name Desulfovibrio oceani sp. nov. is proposed with the two subspecies D. oceani subsp. oceani (type strain, I.8.1T = DSM 21390T = JCM 15970T) and D. oceani subsp. galateae (type strain, I.9.1T = DSM 21391T = JCM 15971T)....

  7. Clonal Streptococcus equi subsp. zooepidemicus post breeding endometritis in thoroughbred broodmares

    DEFF Research Database (Denmark)

    Christoffersen, Mette; Söderlind, Maja; Rydemann Rudefalk, Sofia

    Streptococcus equi subsp. zooepidemicus is one of the most commonly isolated pathogens from the uterus of mares with infectious endometritis. Its ability to cause chronic latent infection by residing deep within the endometrial tissue has previously been described. The aim of the study was to inv......Streptococcus equi subsp. zooepidemicus is one of the most commonly isolated pathogens from the uterus of mares with infectious endometritis. Its ability to cause chronic latent infection by residing deep within the endometrial tissue has previously been described. The aim of the study...... was to investigate whether clonal or genetically distinct S. zooepidemicus strains isolated from mares with endometritis were associated with mare risk factors and the outcome of natural cover. Uterine swabs were obtained from mares with intrauterine fluid after natural cover (n=31) at thoroughbred stud farms...... in Australia. Fifty two percent of the mares (n=16) were diagnosed with infectious endometritis, and S.zooepidemicus was isolated in 81% (n=13) of these mares. Up to four S. zooepidemicus isolates were selected from each mare with growth of S. zooepidemicus and isolates from an additional five mares were...

  8. Multilocus Sex Determination Revealed in Two Populations of Gynodioecious Wild Strawberry, Fragaria vesca subsp. bracteata.

    Science.gov (United States)

    Ashman, Tia-Lynn; Tennessen, Jacob A; Dalton, Rebecca M; Govindarajulu, Rajanikanth; Koski, Matthew H; Liston, Aaron

    2015-10-19

    Gynodioecy, the coexistence of females and hermaphrodites, occurs in 20% of angiosperm families and often enables transitions between hermaphroditism and dioecy. Clarifying mechanisms of sex determination in gynodioecious species can thus illuminate sexual system evolution. Genetic determination of gynodioecy, however, can be complex and is not fully characterized in any wild species. We used targeted sequence capture to genetically map a novel nuclear contributor to male sterility in a self-pollinated hermaphrodite of Fragaria vesca subsp. bracteata from the southern portion of its range. To understand its interaction with another identified locus and possibly additional loci, we performed crosses within and between two populations separated by 2000 km, phenotyped the progeny and sequenced candidate markers at both sex-determining loci. The newly mapped locus contains a high density of pentatricopeptide repeat genes, a class commonly involved in restoration of fertility caused by cytoplasmic male sterility. Examination of all crosses revealed three unlinked epistatically interacting loci that determine sexual phenotype and vary in frequency between populations. Fragaria vesca subsp. bracteata represents the first wild gynodioecious species with genomic evidence of both cytoplasmic and nuclear genes in sex determination. We propose a model for the interactions between these loci and new hypotheses for the evolution of sex determining chromosomes in the subdioecious and dioecious Fragaria. Copyright © 2015 Ashman et al.

  9. Cadmium tolerant characteristic of a newly isolated Lactococcus lactis subsp. lactis.

    Science.gov (United States)

    Sheng, Yao; Wang, Ying; Yang, Xuan; Zhang, Boyang; He, Xiaoyun; Xu, Wentao; Huang, Kunlun

    2016-12-01

    Environmental contamination caused by heavy metals poses a major threat to the wildlife and human health for their toxicity and intrinsically persistent nature. Some specific food grade bacteria have properties that enable them to eliminate heavy metals from food and water. Lactococcus lactis subsp. lactis, newly isolated from pickles, is a cadmium (Cd) tolerant bacteria. Cd resistant properties of the lactis was evaluated under different Cd stresses. Cd accumulation in different cellular parts was determined by ICP-MS and cell morphology changes were measured by SEM-EDS and TEM-EDS. In addition, functional groups associated with Cd resistance were detected by infrared spectroscopic analysis. The results indicated that Cd mainly accumulated in the cell surface structures including cytoderm and cytomembrane. Functional groups such as OH and NH 2 in the cell surface played essential roles in Cd biosorption. The elements of O, P, S, and N of polysaccharide, membrane protein and phosphatidate in the cell surface structures might be responsible for Cd biosorption for their strong electronegativity. This study indicated that ultrastructural analysis can be a supplemental method to study heavy metal resistance mechanism of microorganism and the newly isolated lactococcus lactis subsp. lactis has great potential to be applied to decontamination of heavy metals. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Transcriptomic profile of aguR deletion mutant of Lactococcus lactis subsp. cremoris CECT 8666

    Directory of Open Access Journals (Sweden)

    Beatriz del Rio

    2015-12-01

    Full Text Available Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14 is a dairy strain that catabolizes agmatine (a decarboxylated derivative of arginine into the biogenic amine putrescine by the agmatine deiminase (AGDI pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are responsible for the deamination of agmatine to putrescine and are co-transcribed as a single policistronic mRNA forming the catabolic operon aguBDAC [1]. aguR encodes a transmembrane protein that functions as a one-component signal transduction system that senses the agmatine concentration of the medium and accordingly regulates the transcription of aguBDAC [2], which is also transcriptionally regulated by carbon catabolic repression (CCR via glucose, but not by other sugars such as lactose and galactose [1,3]. Here we report the transcriptional profiling of the aguR gene deletion mutant (L. lactis subsp. cremoris CECT 8666 ∆aguR [2] compared to the wild type strain, both grown in M17 medium with galactose as carbon source and supplemented with agmatine. The transcriptional profiling data of AguR-regulated genes were deposited in the Gene Expression Omnibus (GEO database under accession no. GSE59514.

  11. Essential oils composition of Periploca laevigata Aiton subsp. angustifolia (Labill.) Markgraf (Apocynaceae-Periplocoideae).

    Science.gov (United States)

    Zito, P; Sajeva, M; Bruno, M; Rosselli, S; Maggio, A; Senatore, F

    2013-01-01

    The essential oil of roots, branches, leaves, flowers and fruits of Periploca laevigata Aiton subsp. angustifolia (Apocynaceae) from Lampedusa Island has been obtained by hydrodistillation and its composition analysed. The analyses allowed the identification and quantification of 86 volatile compounds. Branches showed the higher diversity with 57 compounds followed by fruits with 33, roots with 23, flowers with 16 and leaves with six compounds, respectively. In the matrices examined three constituents, heneicosane, docosane and tricosane are in common, although with different percentages. At least the most abundant compounds found in the matrices have been reported to have several biological activities. 2-Hydroxy-4-methoxybenzaldehyde identified in the roots as the most abundant component (70.7%) and present with 8.3% in the branches is a potent tyrosinase inhibitor present in several African medicinal plants, and thus being used as an ingredient in cosmetic and other medicinal products, primarily in relation to hyperpigmentation. Among the compounds identified, several play a role as semiochemicals for many animals, and 28 allomones, 43 pheromones, 21 kairomones have been identified. P. laevigata subsp. angustifolia in Lampedusa Island is host to a community of visitors, and the possible ecological role of the volatiles found is briefly discussed.

  12. Genome Analysis and Characterisation of the Exopolysaccharide Produced by Bifidobacterium longum subsp. longum 35624™.

    Directory of Open Access Journals (Sweden)

    Friedrich Altmann

    Full Text Available The Bifibobacterium longum subsp. longum 35624™ strain (formerly named Bifidobacterium longum subsp. infantis is a well described probiotic with clinical efficacy in Irritable Bowel Syndrome clinical trials and induces immunoregulatory effects in mice and in humans. This paper presents (a the genome sequence of the organism allowing the assignment to its correct subspeciation longum; (b a comparative genome assessment with other B. longum strains and (c the molecular structure of the 35624 exopolysaccharide (EPS624. Comparative genome analysis of the 35624 strain with other B. longum strains determined that the sub-speciation of the strain is longum and revealed the presence of a 35624-specific gene cluster, predicted to encode the biosynthetic machinery for EPS624. Following isolation and acid treatment of the EPS, its chemical structure was determined using gas and liquid chromatography for sugar constituent and linkage analysis, electrospray and matrix assisted laser desorption ionization mass spectrometry for sequencing and NMR. The EPS consists of a branched hexasaccharide repeating unit containing two galactose and two glucose moieties, galacturonic acid and the unusual sugar 6-deoxy-L-talose. These data demonstrate that the B. longum 35624 strain has specific genetic features, one of which leads to the generation of a characteristic exopolysaccharide.

  13. Experimental Inoculation of BFDV-Positive Budgerigars (Melopsittacus undulatus with Two Mycobacterium avium subsp. avium Isolates

    Directory of Open Access Journals (Sweden)

    Aleksandra Ledwoń

    2014-01-01

    Full Text Available Beak and feather disease virus- (BFDV- positive (naturally infected but clinically healthy budgerigars (Melopsittacus undulatus were inoculated with two isolates of Mycobacterium avium subsp. avium isolated from naturally infected golden pheasant (Chrysolophus pictus and peafowl (Pavo cristatus. During a period of more than two months after inoculation, samples of cloacal and crop swabs, faeces, and blood were obtained for BFDV and Mycobacterium avium testing with PCR. Birds were euthanized nine weeks after inoculation. All infected budgerigars developed signs typical of mycobacteriosis, but more advanced clinical and pathological changes were visible in the group infected with the pheasant isolate. Only a few cloacal and crop swab samples were positive for Mycobacterium avium subsp. avium despite advanced pathological changes in the internal organs. In the groups infected with mycobacterium isolates the frequency of BFDV-positive samples was higher than in the control group. In the infected groups the frequency of BFDV was substantially higher in the cloacal swabs of birds inoculated with the pheasant isolate than in the peafowl-isolate-infected group.

  14. Experimental inoculation of BFDV-positive budgerigars (Melopsittacus undulatus) with two Mycobacterium avium subsp. avium isolates.

    Science.gov (United States)

    Ledwoń, Aleksandra; Sapierzyński, Rafał; Augustynowicz-Kopeć, Ewa; Szeleszczuk, Piotr; Kozak, Marcin

    2014-01-01

    Beak and feather disease virus- (BFDV-) positive (naturally infected) but clinically healthy budgerigars (Melopsittacus undulatus) were inoculated with two isolates of Mycobacterium avium subsp. avium isolated from naturally infected golden pheasant (Chrysolophus pictus) and peafowl (Pavo cristatus). During a period of more than two months after inoculation, samples of cloacal and crop swabs, faeces, and blood were obtained for BFDV and Mycobacterium avium testing with PCR. Birds were euthanized nine weeks after inoculation. All infected budgerigars developed signs typical of mycobacteriosis, but more advanced clinical and pathological changes were visible in the group infected with the pheasant isolate. Only a few cloacal and crop swab samples were positive for Mycobacterium avium subsp. avium despite advanced pathological changes in the internal organs. In the groups infected with mycobacterium isolates the frequency of BFDV-positive samples was higher than in the control group. In the infected groups the frequency of BFDV was substantially higher in the cloacal swabs of birds inoculated with the pheasant isolate than in the peafowl-isolate-infected group.

  15. Persistence of Mycobacterium avium subsp. paratuberculosis at a Farm-Scale Biogas Plant Supplied with Manure from Paratuberculosis-Affected Dairy Cattle▿

    Science.gov (United States)

    Slana, I.; Pribylova, R.; Kralova, A.; Pavlik, I.

    2011-01-01

    In this study, products from all steps of anaerobic digestion at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle were examined and quantified for the presence of the causal agent of paratuberculosis, Mycobacterium avium subsp. paratuberculosis, using culture and quantitative real-time PCR (qPCR). Viable M. avium subsp. paratuberculosis cells were detected using culture in fermentors for up to 2 months; the presence of M. avium subsp. paratuberculosis DNA (101 cells/g) was demonstrated in all anaerobic fermentors and digestate 16 months after initiation of work at a biogas plant, using IS900 qPCR. F57 qPCR was able to detect M. avium subsp. paratuberculosis DNA (102 cells/g) at up to 12 months. According to these results, a fermentation process that extended beyond 2 months removed all viable M. avium subsp. paratuberculosis cells and therefore rendered its product M. avium subsp. paratuberculosis free. However, M. avium subsp. paratuberculosis DNA was found during all the examined periods (more than 1 year), which could be explained by either residual DNA being released from dead cells or by the presence of viable cells whose amount was under the limit of cultivability. As the latter hypothesis cannot be excluded, the safety of the final products of digestion used for fertilization or animal bedding cannot be defined, and further investigation is necessary to confirm or refute this risk. PMID:21398476

  16. Derivation of Mutants of Erwinia carotovora subsp. betavasculorum Deficient in Export of Pectolytic Enzymes with Potential for Biological Control of Potato Soft Rot

    Science.gov (United States)

    Costa, José M.; Loper, Joyce E.

    1994-01-01

    Erwinia carotovora subsp. betavasculorum Ecb168 produces an antibiotic(s) that suppresses growth of the related bacterium Erwinia carotovora subsp. carotovora in culture and in wounds of potato tubers. Strain Ecb168 also produces and secretes pectolytic enzymes and causes a vascular necrosis and root rot of sugar beet. Genes (out) involved in secretion of pectolytic enzymes by Ecb168 were localized to two HindIII fragments (8.5 and 10.5 kb) of Ecb168 genomic DNA by hybridization to the cloned out region of E. carotovora subsp. carotovora and by complementation of Out- mutants of E. carotovora subsp. carotovora. Out- mutants of Ecb168, which did not secrete pectate lyase into the culture medium, were obtained when deletions internal to either HindIII fragment were introduced into the genome of Ecb168 through marker exchange mutagenesis. Out- mutants of Ecb168 were complemented to the Out+ phenotype by introduction of the corresponding cloned HindIII fragment. Out- mutants of Ecb168 were less virulent than the Out+ parental strain on potato tubers. Strain Ecb168 and Out- derivatives inhibited the growth of E. carotovora subsp. carotovora in culture, indicating that the uncharacterized antibiotic(s) responsible for antagonism was exported through an out-independent mechanism. Strain Ecb168 and Out- derivatives reduced the establishment of large populations of E. carotovora subsp. carotovora in wounds of potato tubers and suppressed tuber soft rot caused by E. carotovora subsp. carotovora. PMID:16349316

  17. Isolation of Bartonella henselae and Two New Bartonella Subspecies, Bartonella koehlerae Subspecies boulouisii subsp. nov. and Bartonella koehlerae Subspecies bothieri subsp. nov. from Free-Ranging Californian Mountain Lions and Bobcats

    Science.gov (United States)

    Chomel, Bruno B.; Molia, Sophie; Kasten, Rickie W.; Borgo, Gina M.; Stuckey, Matthew J.; Maruyama, Soichi; Chang, Chao-chin; Haddad, Nadia; Koehler, Jane E.

    2016-01-01

    Domestic cats are the natural reservoir of Bartonella henselae, B. clarridgeiae and B. koehlerae. To determine the role of wild felids in the epidemiology of Bartonella infections, blood was collected from 14 free-ranging California mountain lions (Puma concolor) and 19 bobcats (Lynx rufus). Bartonella spp. were isolated from four (29%) mountain lions and seven (37%) bobcats. These isolates were characterized using growth characteristics, biochemical reactions, molecular techniques, including PCR-RFLP of selected genes or interspacer region, pulsed-field gel electrophoresis (PFGE), partial sequencing of several genes, and DNA-DNA hybridization. Two isolates were identical to B. henselae genotype II. All other isolates were distinguished from B. henselae and B. koehlerae by PCR-RFLP of the gltA gene using endonucleases HhaI, TaqI and AciI, with the latter two discriminating between the mountain lion and the bobcat isolates. These two novel isolates displayed specific PFGE profiles distinct from B. henselae, B. koehlerae and B. clarridgeiae. Sequences of amplified gene fragments from the three mountain lion and six bobcat isolates were closely related to, but distinct from, B. henselae and B. koehlerae. Finally, DNA-DNA hybridization studies demonstrated that the mountain lion and bobcat strains are most closely related to B. koehlerae. We propose naming the mountain lion isolates B. koehlerae subsp. boulouisii subsp. nov. (type strain: L-42-94), and the bobcat isolates B. koehlerae subsp. bothieri subsp. nov. (type strain: L-17-96), and to emend B. koehlerae as B. koehlerae subsp. koehlerae. The mode of transmission and the zoonotic potential of these new Bartonella subspecies remain to be determined. PMID:26981874

  18. Isolation of Bartonella henselae and Two New Bartonella Subspecies, Bartonellakoehlerae Subspecies boulouisii subsp. nov. and Bartonella koehlerae Subspecies bothieri subsp. nov. from Free-Ranging Californian Mountain Lions and Bobcats.

    Science.gov (United States)

    Chomel, Bruno B; Molia, Sophie; Kasten, Rickie W; Borgo, Gina M; Stuckey, Matthew J; Maruyama, Soichi; Chang, Chao-Chin; Haddad, Nadia; Koehler, Jane E

    2016-01-01

    Domestic cats are the natural reservoir of Bartonella henselae, B. clarridgeiae and B. koehlerae. To determine the role of wild felids in the epidemiology of Bartonella infections, blood was collected from 14 free-ranging California mountain lions (Puma concolor) and 19 bobcats (Lynx rufus). Bartonella spp. were isolated from four (29%) mountain lions and seven (37%) bobcats. These isolates were characterized using growth characteristics, biochemical reactions, molecular techniques, including PCR-RFLP of selected genes or interspacer region, pulsed-field gel electrophoresis (PFGE), partial sequencing of several genes, and DNA-DNA hybridization. Two isolates were identical to B. henselae genotype II. All other isolates were distinguished from B. henselae and B. koehlerae by PCR-RFLP of the gltA gene using endonucleases HhaI, TaqI and AciI, with the latter two discriminating between the mountain lion and the bobcat isolates. These two novel isolates displayed specific PFGE profiles distinct from B. henselae, B. koehlerae and B. clarridgeiae. Sequences of amplified gene fragments from the three mountain lion and six bobcat isolates were closely related to, but distinct from, B. henselae and B. koehlerae. Finally, DNA-DNA hybridization studies demonstrated that the mountain lion and bobcat strains are most closely related to B. koehlerae. We propose naming the mountain lion isolates B. koehlerae subsp. boulouisii subsp. nov. (type strain: L-42-94), and the bobcat isolates B. koehlerae subsp. bothieri subsp. nov. (type strain: L-17-96), and to emend B. koehlerae as B. koehlerae subsp. koehlerae. The mode of transmission and the zoonotic potential of these new Bartonella subspecies remain to be determined.

  19. Efficacy of florfenicol for control of mortality with Francisella noatunensis subsp. orientalis in Nile tilapia, oreochromis niloticus (L.)

    Science.gov (United States)

    Francisella noatunensis subsp. orientalis (Fno) (syn. F. asiatica) is an emergent Gram-negative facultative intracellular bacterium. Although it is considered one of the most pathogenic bacteria in fish, there are no commercially available treatments of vaccines. The objective of this project was ...

  20. De Roggelelie [Lilium bulbiferum L. subsp. croceum (Chaix) Baker] nog steeds in Drenthe en Noordwest-Duitsland

    NARCIS (Netherlands)

    Bos, Fred

    1989-01-01

    Lilium bulbiferum subsp. croceum has been refound both in Northern Germany and in the Netherlands. The German find is described in more detail, because it concerns a hitherto unknown habitat, viz. a former sand dune area in the valley of the river Ems. In the Netherlands the plant was found again in

  1. Molecular Analyses of the Lactococcin A Gene Cluster from Lactococcus lactis subsp. lactis Biovar Diacetylactis WM4

    NARCIS (Netherlands)

    Stoddard, Gary W.; Petzel, James P.; Belkum, Marco J. van; Kok, Jan; McKay, Larry L.

    1992-01-01

    The genes responsible for bacteriocin production and immunity in Lactococcus lactis subsp. lactis biovar diacetylactis WM4 were localized and characterized by DNA restriction fragment deletion, subcloning, and nucleotide sequence analysis. The nucleotide sequence of a 5.6-kb AvaII restriction

  2. MOLECULAR ANALYSES OF THE LACTOCOCCIN-A GENE-CLUSTER FROM LACTOCOCCUS-LACTIS SUBSP LACTIS BIOVAR DIACETYLACTIS WM4

    NARCIS (Netherlands)

    STODDARD, GW; PETZEL, JP; VANBELKUM, MJ; KOK, J; MCKAY, LL

    The genes responsible for bacteriocin production and immunity in Lactococcus lactis subsp. lactis biovar diacetylactis WM4 were localized and characterized by DNA restriction fragment deletion, subcloning, and nucleotide sequence analysis. The nucleotide sequence of a 5.6-kb AvaII restriction

  3. Divergent cellular responses during asymptomatic subclinical and clinical states of disease in cows naturally infected with Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Infection of the host with Mycobacterium avium subsp. paratuberculosis (MAP) results in a chronic and progressive enteritis that traverses both subclinical and clinical stages. The mechanism(s) for the shift from asymptomatic subclinical disease state to advanced clinical disease are not fully under...

  4. Analysis of Mycobacterium avium subsp. paratuberculosis mutant libraries reveals loci-dependent transcription biases and strategies to novel mutant discovery

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne’s disease in ruminants and it has been implicated as a cause of Crohn’s disease in humans. The generation of comprehensive random mutant banks by transposon mutagenesis is a fundamental wide genomic technology utilized...

  5. Antigenicity of recombinant maltose binding protein-Mycobacterium avium subsp. paratuberculosis fusion proteins with and without factor Xa cleaving

    Science.gov (United States)

    Mycobacterium avium subsp paratuberculosis (MAP) causes Johne’s disease (JD) in ruminants. Proteomic studies have shown that MAP expresses certain proteins when exposed to in vitro physiological stress conditions similar to the conditions experienced within a host during natural infection. Such prot...

  6. Evaluation of two mutants of Mycobacterium avium subsp. paratuberculosis as candidates for a live attenuated vaccine for Johne's disease

    Science.gov (United States)

    Efforts to control Johne’s disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (Map), has been difficult because of a lack of an effective vaccine. To address this problem we examined the potential of targeted gene disruption as a method to develop candidate vaccines with impaired c...

  7. UV Light Inactivation of Mycobacterium avium subsp. paratuberculosis in Milk as Assessed by FASTPlaqueTB Phage Assay and Culture▿

    Science.gov (United States)

    Altic, Leslie C.; Rowe, Michael T.; Grant, Irene R.

    2007-01-01

    UV light inactivation of Mycobacterium avium subsp. paratuberculosis in Middlebrook 7H9 broth and whole and semiskim milk was investigated using a laboratory-scale UV machine that incorporated static mixers within UV-penetrable pipes. UV treatment proved to be less effective in killing M. avium subsp. paratuberculosis suspended in milk (0.5- to 1.0-log10 reduction per 1,000 mJ/ml) than that suspended in Middlebrook 7H9 broth (2.5- to 3.3-log10 reduction per 1,000 mJ/ml). The FASTPlaqueTB phage assay provided more rapid enumeration of surviving M. avium subsp. paratuberculosis (within 24 h) than culture on Herrold's egg yolk medium (6 to 8 weeks). Despite the fact that plaque counts were consistently 1 to 2 log10 lower than colony counts throughout the study, UV inactivation rates for M. avium subsp. paratuberculosis derived using the phage assay and culture results were not significantly different (P = 0.077). PMID:17435001

  8. UV light inactivation of Mycobacterium avium subsp. paratuberculosis in milk as assessed by FASTPlaqueTB phage assay and culture.

    Science.gov (United States)

    Altic, Leslie C; Rowe, Michael T; Grant, Irene R

    2007-06-01

    UV light inactivation of Mycobacterium avium subsp. paratuberculosis in Middlebrook 7H9 broth and whole and semiskim milk was investigated using a laboratory-scale UV machine that incorporated static mixers within UV-penetrable pipes. UV treatment proved to be less effective in killing M. avium subsp. paratuberculosis suspended in milk (0.5- to 1.0-log(10) reduction per 1,000 mJ/ml) than that suspended in Middlebrook 7H9 broth (2.5- to 3.3-log(10) reduction per 1,000 mJ/ml). The FASTPlaqueTB phage assay provided more rapid enumeration of surviving M. avium subsp. paratuberculosis (within 24 h) than culture on Herrold's egg yolk medium (6 to 8 weeks). Despite the fact that plaque counts were consistently 1 to 2 log(10) lower than colony counts throughout the study, UV inactivation rates for M. avium subsp. paratuberculosis derived using the phage assay and culture results were not significantly different (P = 0.077).

  9. Analysis of the exopolysaccharides produced by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 grown in continuous culture on glucose and fructose.

    NARCIS (Netherlands)

    Grobben, G.J.; Casteren, van W.H.M.; Schols, H.A.; Oosterveld, A.; Sala, G.; Smith, M.R.; Sikkema, J.; Bont, de J.A.M.

    1997-01-01

    The exopolysaccharides produced by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772 grown in defined medium were investigated. At equal cell densities, the strain produced 95 mg l−1 exopolysaccharides with glucose and 30 mg l−1 with fructose as the carbohydrate source. High-performance

  10. Increased Production of Hydrogen Peroxide by Lactobacillus delbrueckii subsp. bulgaricus upon Aeration: Involvement of an NADH Oxidase in Oxidative Stress

    Science.gov (United States)

    Marty-Teysset, C.; de la Torre, F.; Garel, J.-R.

    2000-01-01

    The growth of Lactobacillus delbrueckii subsp. bulgaricus (L. delbrueckii subsp. bulgaricus) on lactose was altered upon aerating the cultures by agitation. Aeration caused the bacteria to enter early into stationary phase, thus reducing markedly the biomass production but without modifying the maximum growth rate. The early entry into stationary phase of aerated cultures was probably related to the accumulation of hydrogen peroxide in the medium. Indeed, the concentration of hydrogen peroxide in aerated cultures was two to three times higher than in unaerated ones. Also, a similar shift from exponential to stationary phase could be induced in unaerated cultures by adding increasing concentrations of hydrogen peroxide. A significant fraction of the hydrogen peroxide produced by L. delbrueckii subsp. bulgaricus originated from the reduction of molecular oxygen by NADH catalyzed by an NADH:H2O2 oxidase. The specific activity of this NADH oxidase was the same in aerated and unaerated cultures, suggesting that the amount of this enzyme was not directly regulated by oxygen. Aeration did not change the homolactic character of lactose fermentation by L. delbrueckii subsp. bulgaricus and most of the NADH was reoxidized by lactate dehydrogenase with pyruvate. This indicated that NADH oxidase had no (or a very small) energetic role and could be involved in eliminating oxygen. PMID:10618234

  11. Draft Genome Sequence of Lactobacillus delbrueckii subsp. bulgaricus CFL1, a Lactic Acid Bacterium Isolated from French Handcrafted Fermented Milk

    OpenAIRE

    Meneghel, Julie; Dugat-Bony, Eric; Irlinger, Fran?oise; Loux, Valentin; Vidal, Marie; Passot, St?phanie; B?al, Catherine; Layec, S?verine; Fonseca, Fernanda

    2016-01-01

    Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) is a lactic acid bacterium widely used for the production of yogurt and cheeses. Here, we report the genome sequence of L. bulgaricus CFL1 to improve our knowledge on its stress-induced damages following production and end-use processes.

  12. Survival of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus in the Terminal Ileum of Fistulated Göttingen Minipigs

    Science.gov (United States)

    Lick, Sonja; Drescher, Karsten; Heller, Knut J.

    2001-01-01

    The ability of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus administered in yogurt to survive the passage through the upper gastrointestinal tract was investigated with Göttingen minipigs that were fitted with ileum T-cannulas. After ingestion of yogurt containing viable microorganisms, ileostomy samples were collected nearly every hour beginning 3 h after food uptake. Living L. delbrueckii subsp. bulgaricus and S. thermophilus were detected in the magnitude of 106 to 107 per gram of intestinal contents (wet weight) in all animals under investigation. A calculation of the minimum amount of surviving bacteria that had been administered is presented. Total DNA extracted from ileostomy samples was subjected to PCR, which was species specific for L. delbrueckii and S. thermophilus and subspecies specific for L. delbrueckii subsp. bulgaricus. All three bacterial groups could be detected by PCR after yogurt uptake but not after uptake of a semisynthetic diet. One pig apparently had developed an endogenous L. delbrueckii flora. When heat-treated yogurt was administered, L. delbrueckii was detected in all animals. S. thermophilus or L. delbrueckii subsp. bulgaricus was not detected, indicating that heat-inactivated cells and their DNAs had already been digested and their own L. delbrueckii flora had been stimulated for growth. PMID:11526016

  13. Mediation of host immune responses after immunization of neonatal calves with a heat-killed Mycobacterium avium subsp. paratuberculosis vaccine

    Science.gov (United States)

    A major drawback of current whole-cell vaccines for Mycobacterium avium subsp. paratuberculosis(MAP) is the interference with diagnostic tests for bovine tuberculosis and paratuberculosis. The current study was designed to explore effects of immunization with a heat-killed whole cell vaccine (Mycop...

  14. De Novo whole genome sequence of Xylella fastidiosa subsp. multiplex strain BB01 from blueberry in Georgia, USA

    Science.gov (United States)

    This study reports a de novo assembled draft genome sequence of Xylella fastidiosa subsp. multiplex strain BB01 causing blueberry bacterial leaf scorch in Georgia, USA. The BB01 genome is 2,517,579 bp with a G+C content of 51.8% and 2,943 open reading frames (ORFs) and 48 RNA genes....

  15. Isolation and characterization of Campylobacter jejuni subsp jejuni from macaroni penguins (Eudyptes chrysolophus) in the subantarctic region

    DEFF Research Database (Denmark)

    Broman, T.; Bergstrom, S.; On, Stephen L.W.

    2000-01-01

    On Bird Island, South Georgia, albatrosses (n = 140), penguins (n = 100), and fur seals (n = 206) were sampled for Campylobacter jejuni. C. jejuni subsp. jejuni was recovered from three macaroni penguins (Eudyptes chrysolophus). These isolates, the first reported for the subantarctic region, showed...

  16. Persistence of Mycobacterium avium subsp. paratuberculosis in soil, crops, and ensiled feed following manure spreading on infected dairy farms.

    Science.gov (United States)

    Fecteau, Marie-Eve; Hovingh, Ernest; Whitlock, Robert H; Sweeney, Raymond W

    2013-11-01

    The goal of this study was to determine the persistence of Mycobacterium avium subsp. paratuberculosis (MAP) in soil, crops, and ensiled feeds following manure spreading. This bacterium was often found in soil samples, but less frequently in harvested feeds and silage. Spreading of manure on fields used for crop harvest is preferred to spreading on grazing pastures.

  17. Genome Sequence of the Melanin-Producing Extremophile Aeromonas salmonicida subsp. pectinolytica Strain 34melT.

    Science.gov (United States)

    Pavan, M Elisa; Pavan, Esteban E; López, Nancy I; Levin, Laura; Pettinari, M Julia

    2013-09-12

    The genome of Aeromonas salmonicida subsp. pectinolytica strain 34mel(T), isolated from a heavily polluted river, contains several genomic islands and putative virulence genes. The identification of genes involved in resistance to different kinds of stress sheds light on the mechanisms used by this strain to thrive in an extreme environment.

  18. ZAP-70, CTLA-4, and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Paratuberculosis is a chronic intestinal disease of ruminant animals caused by Mycobacterium avium subsp. paratuberculosis (MAP). A hallmark of paratuberculosis is a transition from a cell-mediated Th1 type response to a humoral Th2 response with the progression of disease from a subclinical to clin...

  19. Immunization with a DNA Vaccine Cocktail Induces a Th1 Response and Protects Mice Against Mycobacterium avium subsp. paratuberculosis Challenge

    Science.gov (United States)

    Several novel antigens of Mycobacterium avium subsp. paratuberculosis have been studied as vaccine components and their immunogenicity has been evaluated. Previously, we reported that 85 antigen complex (85A, 85B, and 85C), superoxide dismutase (SOD), and 35kDa protein could induce significant lymph...

  20. Comparison of specificity and sensitivity of immunochemical and molecular techniques for determination of Clavibacter michiganensis subsp. michiganensis

    Czech Academy of Sciences Publication Activity Database

    Kokošková, B.; Mráz, Ivan; Fousek, Jan

    2010-01-01

    Roč. 55, č. 3 (2010), s. 239-244 ISSN 0015-5632 R&D Projects: GA MZe QH71229 Institutional research plan: CEZ:AV0Z50510513 Keywords : Clavibacter michiganensis subsp. michiganensis (Cmm) * PCR * ELISA * IFAS Subject RIV: EF - Botanics Impact factor: 0.977, year: 2010

  1. Draft genome sequence of Streptococcus equi subsp. zooepidemicus strain S31A1, isolated from equine infectious endometritis

    DEFF Research Database (Denmark)

    da Piedade, Isabelle; Skive, Bolette; Christensen, Henrik

    2013-01-01

    We present the draft genome sequence of Streptococcus equi subsp. zooepidemicus S31A1, a strain isolated from equine infectious endometritis in Denmark. Comparative analyses of this genome were done with four published reference genomes: S. zooepidemicus strains MGCS10565, ATCC 35246, and H70 and S...

  2. Draft Genome Sequences of Four Salmonella enterica subsp. enterica Serovar Enteritidis Strains Implicated in Infections of Avian and Human Hosts

    KAUST Repository

    An, Ran

    2018-01-24

    Salmonella enterica subsp. enterica serovar Enteritidis is a wide-host-range pathogen. Occasionally, it is involved in invasive infections, leading to a high mortality rate. Here, we present the draft genome sequences of four S Enteritidis strains obtained from human and avian hosts that had been involved in bacteremia, gastroenteritis, and primary infections.

  3. Transcription profiling of interactions between Lactococcus lactis subsp. cremoris SK11 and Lactobacillus paracasei ATCC 334 during Cheddar cheese simulation.

    Science.gov (United States)

    Desfossés-Foucault, Émilie; LaPointe, Gisèle; Roy, Denis

    2014-05-16

    The starter cultures (Lactococcus sp.) and non-starter lactic acid bacteria (mostly Lactobacillus spp.) are essential to flavor development of Cheddar cheese. The aim of this study was to elucidate the transcriptional interaction between Lactococcus lactis subsp. cremoris SK11 and Lactobacillus paracasei ATCC 334 in mixed cultures during simulated Cheddar cheese manufacture (Pearce activity test) and ripening (slurry). Reverse transcription quantitative PCR (RT-qPCR) was used to quantify the expression of 34 genes common to both bacteria and for eight genes specific to either L. lactis subsp. cremoris SK11 or L. paracasei ATCC 334. The multifactorial analysis (MFA) performed on fold change results for each gene revealed that the genes linked to stress, protein and peptide degradation as well as carbohydrate metabolism of L. paracasei ATCC 334 were especially overexpressed in mixed culture with L. lactis subsp. cremoris SK11 during the ripening simulation. For L. lactis subsp. cremoris SK11, genes coding for amino acid metabolism were more expressed during the cheese manufacture simulation, especially in single culture. These results show how complementary functions of starter and NSLAB contribute to activities useful for flavor development. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum strains

    Directory of Open Access Journals (Sweden)

    Mohammad J Hossain

    2015-08-01

    Full Text Available To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32- 90%, with 2,839 genes within the core genome of B. amyloliquefaciens subsp. plantarum. Comparative genomic analyses of B. amyloliquefaciens strains identified genes that are linked with biological control and colonization of roots and/or leaves, including 73 genes uniquely associated with subsp. plantarum strains that have predicted functions related to signaling, transportation, secondary metabolite production, and carbon source utilization. Although B. amyloliquefaciens subsp. plantarum strains contain gene clusters that encode many different secondary metabolites, only polyketide biosynthetic clusters that encode difficidin and macrolactin are conserved within this subspecies. To evaluate their role in plant pathogen biocontrol, genes involved in secondary metabolite biosynthesis were deleted in B. amyloliquefaciens subsp. plantarum strain, revealing that difficidin expression is critical in reducing the severity of disease, caused by Xanthomonas axonopodis pv. vesicatoria in tomato plants. This study defines genomic features of PGPR strains and links them with biocontrol activity and with host colonization.

  5. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum

    Science.gov (United States)

    Hossain, Mohammad J.; Ran, Chao; Liu, Ke; Ryu, Choong-Min; Rasmussen-Ivey, Cody R.; Williams, Malachi A.; Hassan, Mohammad K.; Choi, Soo-Keun; Jeong, Haeyoung; Newman, Molli; Kloepper, Joseph W.; Liles, Mark R.

    2015-01-01

    To understand the growth-promoting and disease-inhibiting activities of plant growth-promoting rhizobacteria (PGPR) strains, the genomes of 12 Bacillus subtilis group strains with PGPR activity were sequenced and analyzed. These B. subtilis strains exhibited high genomic diversity, whereas the genomes of B. amyloliquefaciens strains (a member of the B. subtilis group) are highly conserved. A pairwise BLASTp matrix revealed that gene family similarity among Bacillus genomes ranges from 32 to 90%, with 2839 genes within the core genome of B. amyloliquefaciens subsp. plantarum. Comparative genomic analyses of B. amyloliquefaciens strains identified genes that are linked with biological control and colonization of roots and/or leaves, including 73 genes uniquely associated with subsp. plantarum strains that have predicted functions related to signaling, transportation, secondary metabolite production, and carbon source utilization. Although B. amyloliquefaciens subsp. plantarum strains contain gene clusters that encode many different secondary metabolites, only polyketide biosynthetic clusters that encode difficidin and macrolactin are conserved within this subspecies. To evaluate their role in plant pathogen biocontrol, genes involved in secondary metabolite biosynthesis were deleted in a B. amyloliquefaciens subsp. plantarum strain, revealing that difficidin expression is critical in reducing the severity of disease, caused by Xanthomonas axonopodis pv. vesicatoria in tomato plants. This study defines genomic features of PGPR strains and links them with biocontrol activity and with host colonization. PMID:26347755

  6. Pyrrolizidine Alkaloids from Symphytum sylvaticum Boiss. subsp. sepulcrale. (Boiss.& Bal.) Greuter & Burdetvar. sepulcrale and Symphytum aintabicum Hub.- Mor. & Wickens

    OpenAIRE

    KURUCU, Semra; KARTAL, Murat

    2002-01-01

    Pyrrolizidine alkaloid (Echimidine-N-oxide) was isolated from Symphytum sylvaticum Boiss. subsp. sepulcrale (Boiss. & Bal.) Greuter & Burdet var. sepulcrale and pyrrolizidine alkaloid (Echimidine) was isolated from Symphytum aintabicum Hub. - Mor. & Wickens. The structures of the isolated compounds were elucidated based on IR, EIMS, 1H, and 13C NMR analysis and also on 2D NMR (COSY, HMBC, HMQC) experiments.

  7. Pectobacterium carotovorum subsp. carotovorum strains show diversity in production of and response to N-acyl homoserine lactones

    NARCIS (Netherlands)

    Jafra, S.; Jalink, H.; Schoor, van der R.; Wolf, van der J.M.

    2006-01-01

    Pectobacterium carotovorum subsp.carotovorum (Pcc) is a plant pathogen, which can cause soft rot in a wide range of plants. A regulatory network controls the synthesis of virulence factors, mainly plant cell wall degrading enzymes, in a cell density dependent manner. Small signalling molecules,

  8. Use of an Individual-based Model to Control Transmission Pathways of Mycobacterium avium Subsp paratuberculosis Infection in Cattle Herds

    NARCIS (Netherlands)

    Al-Mamun, Mohammad A; Smith, Rebecca L; Schukken, Y. H.; Grohn, Y. T.

    2017-01-01

    Johne’s disease (JD) is a chronic enteric disease in cattle caused by Mycobacterium avian subsp. paratuberculosis (MAP). Eradicating JD is a difficult task due to the long incubation period of MAP, inefficient diagnostic tests, and delayed clinical signs. Effective control strategies can help

  9. Use of an Individual-based Model to Control Transmission Pathways of Mycobacterium avium Subsp. paratuberculosis Infection in Cattle Herds

    NARCIS (Netherlands)

    Mamun Hossain, Al Shaikh Abdullah; Smith, R.L.; Schukken, Y.H.; Gröhn, Y.T.

    2017-01-01

    Johne's disease (JD) is a chronic enteric disease in cattle caused by Mycobacterium avian subsp. paratuberculosis (MAP). Eradicating JD is a difficult task due to the long incubation period of MAP, inefficient diagnostic tests, and delayed clinical signs. Effective control strategies can help

  10. Biological control of Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) using Aeromonas phage PAS-1.

    Science.gov (United States)

    Kim, J H; Choresca, C H; Shin, S P; Han, J E; Jun, J W; Park, S C

    2015-02-01

    The potential control efficacy of Aeromonas phage PAS-1 was evaluated against Aeromonas salmonicida subsp. salmonicida infection in rainbow trout (Oncorhynchus mykiss) model in this study. The phage was co-cultured with the virulent A. salmonicida subsp. salmonicida strain AS05 that possesses the type III secretion system (TTSS) ascV gene, and efficient bacteriolytic activity was observed against the bacteria. The administration of PAS-1 in rainbow trout demonstrated that the phage was cleared from the fish within 200 h post-administration, and a temporal neutralizing activity against the phage was detected in the sera of phage-administrated fish. The administration of PAS-1 (multiplicity of infection: 10 000) in A. salmonicida subsp. salmonicida infected rainbow trout model showed notable protective effects, with increased survival rates and mean times to death. These results demonstrated that Aeromonas phage PAS-1 could be considered as an alternative biological control agent against A. salmonicida subsp. salmonicida infections in rainbow trout culture. © 2013 Blackwell Verlag GmbH.

  11. Complete genomic sequences of two salmonella enterica subsp. enterica serogroup C2 (O:6,8) strains from central California

    Science.gov (United States)

    Salmonella enteric subsp. enterica strains RM11060, serotype 6,8:d:-, and RM11065, serotype 6,8:-:e,n,z15, were isolated from environmental sampling in Central California in 2009. We report the complete genome sequences and annotation of these two strains. These genomic sequences are distinct and wi...

  12. Heterologous Gene Expression in Lactococcus lactis subsp. lactis : Synthesis, Secretion, and Processing of the Bacillus subtilis Neutral Protease

    NARCIS (Netherlands)

    Guchte, Maarten van de; Kodde, Jan; Vossen, Jos M.B.M. van der; Kok, Jan; Venema, Gerard

    1990-01-01

    The Bacillus subtilis nprE gene lacking its own promoter sequence was inserted in the lactococcal expression vector pMG36e. Upon introduction of the recombinant plasmid into Lactococcus lactis subsp. lactis strain MG1363, neutral protease activity could be visualized by the appearance of large

  13. Draft genome sequence of Enterobacter cloacae subsp. cloacae strain 08XA1, a fecal bacterium of giant pandas.

    Science.gov (United States)

    Yan, Yue; Zhao, Chuan-Wu; Zhang, Yi-Zheng; Zhang, Zhi-He; Pan, Guang-Lin; Liu, Wen-Wang; Ma, Qing-Yi; Hou, Rong; Tan, Xue-Mei

    2012-12-01

    Enterobacter cloacae, a common pathogenic bacterium, is a Gram-negative bacillus. We analyzed the draft genome of Enterobacter cloacae subsp. cloacae strain 08XA1 from the feces of a giant panda in China. Genes encoding a β-lactamase and efflux pumps, as well as other factors, have been found in the genome.

  14. Biofilm formation and transcriptome analysis of Streptococcus gallolyticus subsp. gallolyticus in response to lysozyme.

    Directory of Open Access Journals (Sweden)

    Imke Grimm

    Full Text Available Streptococcus gallolyticus subsp. gallolyticus is a commensal bacterium of the human gastrointestinal tract, and a pathogen causing infective endocarditis and other biofilm-associated infections via exposed collagen. This study focuses on the characterization of the biofilm formation and collagen adhesion of S. gallolyticus subsp. gallolyticus under different conditions. In this study, it has been observed that the isolate UCN 34 is resistant to 20 mg/ml lysozyme in BHI medium, whereas the strain BAA-2069 builds more biofilm in the presence of lysozyme compared to in a control of BHI without lysozyme. A transcriptome analysis with whole genome microarrays of these two isolates in BHI medium with lysozyme compared to control without lysozyme revealed changes in gene expression levels. In the isolate BAA-2069, 67 genes showed increased expression in the presence of lysozyme, while in the isolate UCN 34, 165 genes showed increased expression and 30 genes showed decreased expression through lysozyme treatment. Products of genes which were higher expressed are in involved in transcription and translation, in cell-wall modification, in hydrogen peroxide resistance and in bacterial immunity. Furthermore, the adhesion ability of different strains of S. gallolyticus subsp. gallolyticus to collagen type I and IV was analyzed. Thereby, we compared the adhesion of 46 human isolates with 23 isolates from animals. It was shown that the adhesion ability depends significantly on whether the isolate was isolated from human or animal. For example, high adhesion ability was observed for strain UCN 34 isolated from an infective endocarditis patient, whereas strain DSM 16831 isolated from koala feces adhered only marginally to collagen. Full genome microarray analysis of these two strains revealed strain-dependent gene expression due to adhesion. The expression of 25 genes of a transposon and 15 genes of a phage region in strain DSM 16831 were increased, which

  15. Biochemical, immunological and toxicological characteristics of the crystal proteins of Bacillus thuringiensis subsp. medellin

    Directory of Open Access Journals (Sweden)

    Sergio Orduz

    1996-04-01

    Full Text Available Characterization of the insecticidal and hemolytic activity of solubilized crystal proteins of Bacillus thuringiensis (Bt subsp. medellin (Btmed was performed and compared to solubilized crystal proteins of isolates 1884 of B. thuringiensis subsp. israelensis (Bti and isolate PG-14 of B. thuringiensis subsp. morrisoni (Btm. In general, at acid pH values solubilization of the Bt crystalline parasporal inclusions (CPI was lower than at alkaline pH. The larvicidal activity demonstrated by the CPI of Btmed indicated that optimal solubilization of CPI takes place at a pH value of 11.3, in Bti at pH values from 5.03 to 11.3 and in Btm at pH values from 9.05 to 11.3. Hemolytic activity against sheep red blood cells was mainly found following extraction at pH 11.3 in all Bt strains tested. Polyacrylamide gel electrophoresis under denaturing conditions revealed that optimal solubilization of the CPI in all Bt strains takes place at the alkaline pH values from 9.05 to 11.3. An enriched preparation of Btmed crystals was obtained, solubilized and crystal proteins were separated on a size exclusion column (Sephacryl S-200. Three main protein peaks were observed on the chromatogram. The first peak had two main proteins that migrate between 90 to 100 kDa. These proteins are apparently not common to other Bt strains isolated to date. The second and third peaks obtained from the size exclusion column yielded polypeptides of 68 and 28-30 kDa, respectively. Each peak independently, showed toxicity against 1st instar Culex quinquefasciatus larvae. Interestingly, combinations of the fractions corresponding to the 68 and 30 kDa protein showed an increased toxicity. These results suggest that the 94 kDa protein is an important component of the Btmed toxins with the highest potency to kill mosquito larvae. When crystal proteins of Bti were probed with antisera raised independently against the three main protein fractions of Btmed, the only crystal protein that showed

  16. Intoxicação espontânea por Ipomoea carnea subsp. fistulosa (Convolvulaceae em bovinos no Pantanal Matogrossense Spontaneous Ipomoea carnea subsp. fistulosa (Convolvulaceae poisoning of cattle in the Brazilian Pantanal

    Directory of Open Access Journals (Sweden)

    Nadia A.B. Antoniassi

    2007-10-01

    Full Text Available Relata-se a intoxicação espontânea por Ipomoea carnea subsp. fistulosa (canudo, algodoeiro em bovinos no Pantanal Matogrossense. As investigações iniciaram após a morte de 12 bovinos, de um rebanho de 500 animais, criados em uma extensa área intensamente infestada por I. carnea subsp. fistulosa com escassa disponibilidade de outra forragem. As mortes ocorreram entres os meses de junho e setembro de 2006. O quadro clínico foi caracterizado por emagrecimento e sinais neurológicos com dificuldade locomotora. Um bovino foi necropsiado sem que se observassem alterações macroscópicas significativas. Histologicamente havia tumefação e vacuolização celular, em neurônios, células acinares pancreáticas, tubulares renais e foliculares da tireóide. Bovinos com quadro clínico similar foram retirados da área invadida por I. carnea subsp. fistulosa e colocadas em áreas com pastagem nativa e de Brachiaria sp. e apresentaram melhora clínica após período de 15 dias.A spontaneous Ipomoea carnea subsp. fistulosa (canudo, algodoeiro poisoning of cattle in the county of Poconé, Brazilian Pantanal, is reported. The investigation began after 12 cattle had died from a flock of 500 animals maintained in an extensive area intensely infested by I. carnea subsp. fistulosa with scarce availability of other fodder plants. The deaths occurred from June to September of 2006. Clinical signs were loss of weight and neurological deficits with hypermetry and incoordination. No significant gross lesions were observed at postmortem examination of one bovine. Histological changes comprised widespread cytoplasmic vacuolation of neurons, cells of the thyroid, kidney and pancreas. Cattle with similar clinical picture, that had been removed from the area invaded by I. carnea subsp. fistulosa and placed into areas with native and Brachiaria sp. pasture, recovered clinically within 15 days.

  17. Identification of expressed genes during infection of chinese cabbage (Brassica rapa subsp. pekinensis) by Plasmodiophora brassicae

    DEFF Research Database (Denmark)

    Sundelin, Thomas; Jensen, Dan Funck; Lübeck, Mette

    2011-01-01

    and that the introns are small. These results show that it is possible to discover new P. brassicae genes from a mixed pool of both plant and pathogen cDNA. The results also revealed that some of the P. brassicae genes expressed in Chinese cabbage (Brassica rapa subsp. pekinensis) were identical to the genes expressed......Plasmodiophora brassicae is an obligate, biotrophic pathogen causing the club-root disease of crucifers. Despite its importance as a plant pathogen, little is known about P. brassicae at the molecular level as most of its life cycle takes place inside the plant host, and axenic culturing...... is impossible. Discovery of genes expressed during infection and gene organization are the first steps toward a better understanding of the pathogen–host interaction. Here, suppression subtractive hybridization was used to search for the P. brassicae genes expressed during plant infection. One-hundred and forty...

  18. Occurrence of Mycobacterium avium subsp. paratuberculosis in milk at dairy cattle farms

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Nielsen, Søren Saxmose

    2012-01-01

    Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in milk for human consumption is a concern due to its possible relationship with Crohn’s disease in humans. Pasteurization effectively reduces the MAP load by four to five logs, but the efficacy depends on the MAP concentration, which...... depends on the prevalence among contributing herds and individuals. Considerable variation of MAP in bulk tank milk (BTM) and individual cow’s milk (IM) is reported, but factors associated with MAP occurrence in milk at farm level have not been described. This study systematically reviewed published...... studies aiming at estimating the occurrence of MAP in on-farm BTM and IM by meta-analysis. A total of 692 articles were identified through electronic databases and initially screened using title and abstract. The quality of the 61 potentially relevant articles was assessed using full text and 31 articles...

  19. The Investigation of Some Bioactive Compounds and Antioxidant Properties of Hawthorn (Crataegus monogyna subsp. monogyna jacq.

    Directory of Open Access Journals (Sweden)

    Serhat KESER

    2014-04-01

    Full Text Available The antioxidant and pharmacological effects of hawthorn have mainly been attributed to the polyphenolic contents. The aim of this research is to determine some bioactive compounds and antioxidant properties of hawthorn aqueous and ethanol extracts of leaves, flowers and ripened fruits. For this purpose, antioxidant activities of extracts were assessed on DPPH and #8226;, ABTS and #8226;+, superoxide scavenging, reducing power and ferrous metal chelating activity assays and phenolic content of extracts was determined by Folin-Cioacalteu and #8217;s reagent. The flavonoids including rutin, apigenin, myricetin, quercetin, naringenin and kaempferol, were identified by HPLC in the hawthorn extract. It was observed the aqueous and ethanol extracts of Crataegus monogyna subsp. monogyna fruits showed highest activity in reducing power and metal chelating activity assays. Additionally, it was determined that the aqueous flower extract showed higher flavonoid content than aqueous leaves extract. [J Intercult Ethnopharmacol 2014; 3(2.000: 51-55

  20. Phenylpropanoids, Sesquiterpenoids and Flavonoids from Pimpinella tragium Vill . subsp. lithophila (Schischkin Tutin

    Directory of Open Access Journals (Sweden)

    Hilal Özbek

    2016-03-01

    Full Text Available A new sesquiterpenoid named germacradiene-6 -O-(6'-O-acetyl-β-D-glucoside (1 and a new flavonol glycoside named rhamnetin-3-O-(2′′-O- β-D-glucopyranosyl- β-D-galactopyranoside (2, along with three known sesquiterpenoids dictamnol (3, radicol (4, germacradiene glucoside (5; three phenylpropanoids 4-methoxy-2-(3-methyloxiranyl-phenyl 2-methylbutanoate (6, 4-methoxy-2-(3-methyloxiranyl-phenyl angelate (7, thellungianin E (8; and a flavonol glycoside platanoside (9 were isolated from the aerial parts of Pimpinella tragium Vill . subsp. lithophila (Schischkin Tutin. Their structures were elucidated by detailed analyses of 1D and 2D NMR, UV, IR and HR-ESI-MS data.

  1. Detection of Mycobacterium avium subsp. paratuberculosis in Milk from Clinically Affected Cows by PCR and culture

    DEFF Research Database (Denmark)

    Giese, Steen Bjørck; Ahrens, Peter

    1999-01-01

    animals. In milk from 5 cows (all faecal culture-positive) we cultivated a few colonies of M. a. paratuberculosis (less than 100 CFU per mi). Milk samples from 2 cows were PCR-positive (both animals were faecal culture-positive, and 1 cow was milk culture positive). One cow was culture......Milk and faecal samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp.paratuberculosis (M. a. paratuberculosis) by culture and PCR. M. a. paratuberculosis was isolated in varied numbers from faeces or intestinal mucosa in 8 of 11......-negative on intestinal mucosa, but culture-positive in milk, and both faeces and milk were negative in culture and PCR from 2 cows. In conclusion the presence of M. a. paratuberculosis could be detected in raw milk by PCR but cultivation of milk was more sensitive in detecting the organism....

  2. Aeromonas salmonicida subsp. salmonicida in the light of its type-three secretion system

    Science.gov (United States)

    Vanden Bergh, Philippe; Frey, Joachim

    2014-01-01

    Aeromonas salmonicida subsp. salmonicida is an important pathogen in salmonid aquaculture and is responsible for the typical furunculosis. The type-three secretion system (T3SS) is a major virulence system. In this work, we review structure and function of this highly sophisticated nanosyringe in A. salmonicida. Based on the literature as well as personal experimental observations, we document the genetic (re)organization, expression regulation, anatomy, putative functional origin and roles in the infectious process of this T3SS. We propose a model of pathogenesis where A. salmonicida induces a temporary immunosuppression state in fish in order to acquire free access to host tissues. Finally, we highlight putative important therapeutic and vaccine strategies to prevent furunculosis of salmonid fish. PMID:24119189

  3. Complete genome sequence of the Pectobacterium carotovorum subsp. carotovorum virulent bacteriophage PM1.

    Science.gov (United States)

    Lim, Jeong-A; Shin, Hakdong; Lee, Dong Hwan; Han, Sang-Wook; Lee, Ju-Hoon; Ryu, Sangryeol; Heu, Sunggi

    2014-08-01

    PM1, a novel virulent bacteriophage that infects Pectobacterium carotovorum subsp. carotovorum, was isolated. Its morphological features were examined by electron microscopy, which indicated that this phage belongs to the family Myoviridae. It has a 55,098-bp genome, including a 2,665-bp terminal repeat. A total of 63 open reading frames (ORFs) were predicted, but only 20 ORFs possessed homology with functional proteins. There is one tRNA coding region, and the GC-content of the genome is 44.9 %. Most ORFs in bacteriophage PM1 showed high homology to enterobacteria phage ΦEcoM-GJ1 and Erwinia phage νB EamM-Y2. Like these bacteriophages, PM1 encodes an RNA polymerase, which is a hallmark of T7-like phages. There is no integrase or repressor, suggesting that PM1 is a virulent bacteriophage.

  4. Structure determination of the neutral exopolysaccharide produced by Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1.

    Science.gov (United States)

    Van Calsteren, Marie-Rose; Gagnon, Fleur; Nishimura, Junko; Makino, Seiya

    2015-09-02

    The neutral exopolysaccharide (NPS) of Lactobacillus delbrueckii subsp. bulgaricus strain OLL1073R-1 was purified and characterized. The molecular mass was 5.0×10(6) g/mol. Sugar and absolute configuration analyses gave the following composition: d-Glc, 1; d-Gal, 1.5. The NPS was also submitted to periodate oxidation followed by borohydride reduction and Smith degradation. Sugar and methylation analyses, (1)H and (13)C nuclear magnetic resonance, and mass spectrometry of the NPS or of its specifically modified products allowed determining the repeating unit sequence: {2)Glc(α1-3)Glc(β1-3)[Gal(β1-4)]Gal(β1-4)Gal(α1-}n. The structure is compared to that of exopolysaccharides produced by other Lactobacillus bulgaricus strains. Copyright © 2015. Published by Elsevier Ltd.

  5. Produksi Asam Laktat oleh Lactobacillus delbrueckii subsp. bulgaricus dengan Sumber Karbon Tetes Tebu

    Directory of Open Access Journals (Sweden)

    Laita Nurjannah

    2017-04-01

    Full Text Available Senyawa asam laktat sangat dibutuhkan di dunia industri. Namun produksi dengan menggunakan mikrob masih menggunakan bahan pangan sebagai substratnya. Alternatif substrat untuk produksi asam laktat  sebagai pengganti penggunaan bahan pangan  sangat diperlukan industri. Tetes tebu merupakan salah satu substrat yang kaya akan sumber  karbon yang dapat digunakan sebagai komponen media pertumbuhan bakteri. Ketersediaannya melimpah dan harganya murah. Tujuan penelitian ini adalah tetes tebu dapat digunakan sebagai alternatif  sumber karbon bakteri Lactobacillus delbrueckii subsp. bulgaricus untuk menghasilkan asam laktat. Langkah penelitian ini meliputi hidrolisis dan detoksifikasi tetes tebu, uji kualitatif gula pereduksi tetes tebu, analisis gula total dengan metode fenol sulfat, penentuan kurva pertumbuhan bakteri, produksi dan ekstraksi asam laktat, serta analisis kualitatif asam laktat dengan menggunakan kromatografi cair kinerja tinggi. Hasil penelitian menunjukkan bahwa tetes tebu dapat digunakan sebagai alternatif sumber karbon. Hal ini terbukti bakteri dapat tumbuh dengan baik ketika media diberi 0.5% tetes tebu. Konsentrasi gula total tetes tebu adalah 1090 g/L. Uji gula pereduksi menunjukkan hasil yang positif untuk uji Selliwanof, uji Benedict, dan uji Barfoed. Pertumbuhan optimum L. delbrueckii subsp. bulgaricus terjadi pada suhu 42°C dengan agitasi 150 rpm. Produksi asam laktat dilakukan selama 24 jam. Kadar asam laktat yang dihasilkan sebesar 2.80% dengan biomassa sel kering sebesar 0.002 g/L dan pH media fermentasi sebesar 4.0. Hasil analisis kualitatif kromatografi cair kinerja tinggi juga menunjukkan bahwa produk dari hasil fermentasi adalah asam laktat. Abstract. Lactic acid is needed as an industrial feed. However, by using a microbial production still uses food material as a substrate. Alternative substrates for the production of lactic acid is needed in industry. Molasses are potential substrates due to the richness in

  6. A model of proteolysis and amino acid biosynthesis for Lactobacillus delbrueckii subsp. bulgaricus in whey.

    Science.gov (United States)

    Liu, Enuo; Zheng, Huajun; Hao, Pei; Konno, Tomonobu; Yu, Yao; Kume, Hisae; Oda, Munehiro; Ji, Zai-Si

    2012-12-01

    Lactobacillus delbrueckii subsp. bulgaricus 2038 (L. bulgaricus 2038) is a bacterium that is used as a starter for dairy products by Meiji Co., Ltd of Japan. Culturing L. bulgaricus 2038 with whey as the sole nitrogen source results in a shorter lag phase than other milk proteins under the same conditions (carbon source, minerals, and vitamins). Microarray results of gene expression revealed characteristics of amino acid anabolism with whey as the nitrogen source and established a model of proteolysis and amino acid biosynthesis for L. bulgaricus. Whey peptides and free amino acids are readily metabolized, enabling rapid entry into the logarithmic growth phase. The oligopeptide transport system is the primary pathway for obtaining amino acids. Amino acid biosynthesis maintains the balance between amino acids required for cell growth and the amount obtained from environment. The interconversion of amino acids is also important for L. bulgaricus 2038 growth.

  7. Soil bacteria Pseudomonas putida and Alcaligenes xylosoxidans subsp. denitrificans inactivate triclosan in liquid and solid substrates.

    Science.gov (United States)

    Meade, M J; Waddell, R L; Callahan, T M

    2001-10-16

    Triclosan is a broad-spectrum antimicrobial agent that has been incorporated into many household and medical products. Bacteria with high levels of triclosan resistance were isolated from compost, water, and soil samples. Two of these bacteria, Pseudomonas putida TriRY and Alcaligenes xylosoxidans subsp. denitrificans TR1, were able to use triclosan as a sole carbon source and clear particulate triclosan from agar. A decrease in triclosan concentration was measured by HPLC within 6 h of inoculation with strain TriRY and 24 h with strain TR1. Bioassays demonstrated that triclosan was inactivated in liquid cultures and/or embedded in plastic by the growth of strain TriRY and strain TR1, permitting the growth of triclosan-sensitive bacteria.

  8. Impact of salt concentration on persistence of Mycobacterium avium subsp. paratuberculosis in Iranian UF white cheese

    Directory of Open Access Journals (Sweden)

    Sh Hanifian

    2013-02-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis is considered as a potential significant public health threat due to its possible association with Crohn’s disease in humans. This is a study aimed to investigate the effect of different salt concentrations on survival of Mycobacterium paratuberculosis during ripening and storage of Iranian ultra-filtrate-white cheese (IUFWC. For this purpose, retentate was inoculated with 2 Log cfu/g of Mycobacterium paratuberculosis. Afterwards, model cheeses were prepared with 2%, 3% and 4% of salt. Quantity of Mycobacterium paratuberculosis was estimated throughout the ripening and storage of IUFWC using F57-quantitative real time PCR (F57-qPCR and culture assay. Along with, the populations of lactic acid bacteria as well as physicochemical properties of cheese samples were determined. According to the results, at the early stage of storage period (1 to 30 days the number of Mycobacterium paratuberculosis was almost constant; however, it was decreased significantly (p

  9. Inactivation of Xanthomonas citri subsp. citri and Effect on Infection of Citrus Canker by Gamma Irradiation

    Directory of Open Access Journals (Sweden)

    Kyung Nam Kim

    2015-06-01

    Full Text Available Citrus canker caused by Xanthomonas citri subsp. citri (Xcc has been quarantined by many countries in the world. Recently, the usage of methyl bromide should be limited, application by gamma irradiation on the agricultural production is raised as an alternative method. In this study, the level of gamma irradiation which could decrease of population of Xcc in the suspension or on the surface of citrus fruit was investigated. The D10 value of Xcc, which is radiation dose required to reduce the number of the microorganism, was 55 and 28 Gy in the suspension and on the surface of citrus fruit, respectively. Furthermore, disease severity was suppressed on the citrus leaves inoculated with Xcc suspension pre-treated with gamma irradiation. Based on this study, it is suggested that Xcc on the citrus fruit could be eradicated by gamma irradiation and the results of this study may be valuable for application of gamma ray in quarantine activity

  10. Apparent prevalence of beef carcasses contaminated with Mycobacterium avium subsp. paratuberculosis sampled from Danish slaughter cattle

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Pozzato, Nicola

    2011-01-01

    Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in beef has been reported as a public health concern because asymptomatically infected cattle may contain MAP in tissues that are used for human consumption. Associations between MAP carcasses contamination and animal characteristics...... such as age, breed, production type, and carcass classification were assessed. Cheek muscles from 501 carcasses were sampled cross-sectionally at a Danish abattoir and tested for presence of viable MAP and MAP DNA by bacterial culture and IS900 realtime PCR, respectively. Cheek muscle tissues from carcasses...... of two dairy cows were positive by culture whereas 4% of the animals were estimated with =10¿CFU/gram muscle based on realtime PCR. Age was found to be associated with carcass contamination with MAP. The observed viable MAP prevalence in beef carcasses was low. However, detection of MAP and MAP DNA...

  11. Enzymatic properties of lipase and characteristics production by Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    el-Sawah, M M; Sherief, A A; Bayoumy, S M

    1995-01-01

    Some properties of an extracellular lipase produced by Lactobacillus delbrueckii subsp. bulgaricus were studied. Maximum enzyme activity was found against olive and butter oil as enzyme substrates. Addition of 9% acacia gum, 0.1% Na-deoxycholate and 0.01 M CaCl2 to the enzyme reaction mixture increased-lipase activity from 5.3 to 14.5 (FFA/mg protein/minute) at pH 6.0 and at 40 degrees C. Maximum lipase production was reached in the presence of glucose as a sole source of carbon, wheat bran as nitrogen source, olive oil as a sole lipid source and butyric acid as fatty acid supporting the growth medium. An initial pH value of the culture medium of 6.0 and a temperature of 35 degrees C gave the highest lipolytic activity.

  12. CD44 enhances macrophage phagocytosis and plays a protective role in Streptococcus equi subsp. zooepidemicus infection.

    Science.gov (United States)

    Fu, Qiang; Wei, Zigong; Xiao, Pingping; Chen, Yaosheng; Liu, Xiaohong

    2017-01-01

    Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is an important pathogen associated with a wide range of diseases in many mammalian species. CD44 is a transmembrane adhesion molecule involved in innate and adaptive immune responses. The aim of this study was to determine the protective role of CD44 during S. zooepidemicus infection. CD44-deficient mice exhibited reduced macrophage accumulation in the bronchoalveolar space and enhanced bacterial outgrowth and dissemination, which resulted in reduced mouse survival. An in vitro analysis revealed that CD44 can directly bind to S. zooepidemicus. Additionally, S. zooepidemicus interacted with macrophage-associated CD44, as reflected by the reduced uptake of S. zooepidemicus by CD44-deficient macrophages. These data suggest that CD44 contributes to effective antibacterial defense during S. zooepidemicus infection, thereby limiting the accompanying injury and death. Copyright © 2016. Published by Elsevier B.V.

  13. Two new species of Lactarius associated with Alnus acuminata subsp. arguta in Mexico.

    Science.gov (United States)

    Montoya, Leticia; Bandala, Victor M; Garay, Edith

    2014-01-01

    In pure stands of Alnus acuminata subsp. arguta trees from Sierra Norte de Puebla (central Mexico) two undescribed ectomycorrhizal species of Lactarius were discovered. Distinction of the two new species is based on morphological characters and supported with phylogenetic analyses of the nuclear ribosomal DNA ITS region and part of the gene that encodes for the second largest subunit of RNA polymerase II (rpb2). The phylogenies inferred recovered the two species in different clades strongly supported by posterior probabilities and bootstrap values. The new Lactarius species are recognized as part of the assemblage of ectomycorrhizal fungi associated with Alnus acuminata. Information about these taxa includes the morphological variation achieved along 16 monitories 2010-2013. Descriptions are provided. They are accompanied by photos including SEM photomicrographs of basidiospores and information on differences between them and other related taxa from Europe and the United States. © 2014 by The Mycological Society of America.

  14. Draft genome of a South African strain of Pectobacterium carotovorum subsp. brasiliense.

    Science.gov (United States)

    Ntushelo, Khayalethu; Mafofo, Joseph

    The draft genome of Pectobacterium carotovorum subsp. brasiliense (Pcb) which causes blackleg of potato was submitted to the NCBI and released with reference number NZ_LGRF00000000.1. The estimated genome size based on the draft genome assembly is 4,820,279bp from 33 contigs ranging in length from 444 to 1,660,019 nucleotides. The genome annotation showed 4250 putative genes, 4114 CDS and 43 pseudo-genes. Three complete rRNA gene species were detected: nine 5S, one 16S and one 23S. Other partial rRNA gene fragments were also identified, nine 16S rRNA and three 23S rRNA. A total of 69 tRNA genes and one ncRNA gene were also annotated in this genome. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  15. Pectobacterium carotovorum subsp. carotovorum – the Causal Agent of Calla Soft Rot in Serbia and Montenegro

    Directory of Open Access Journals (Sweden)

    Milan Ivanović

    2009-01-01

    Full Text Available Bacterial strains were isolated from above- and underground parts of diseased calla plants originating from different localities in Serbia and one locality in Montenegro. They were characterized by studying their pathogenic, cultural, biochemical and physiologicalcharacteristics. All investigated strains caused soft rot of calla leaf stalks, potato slices and aloe leaves, and induced hypersensitive reaction on tobacco. Bacteriological properties of the strains indicated that symptoms on calla plants were caused by Gram-negative, nonfluorescent, oxidase negative, catalase positive and facultatively anaerobic bacterium belonging to the genus Pectobacterium. The investigated strains grew at 37ºC and in 5% NaCl, utilised lactose and trechalose, and produced neither indol nor lecitinase. These results, as well as the characteristic growth on Logan’s differential medium indicated that soft rot of tuber and stem base of calla plants was caused by Pectobacterium carotovorum subsp. carotovorum. This is the first report of this pathogen affecting calla plants in Serbia.

  16. Draft genome of a South African strain of Pectobacterium carotovorum subsp. brasiliense

    Directory of Open Access Journals (Sweden)

    Khayalethu Ntushelo

    Full Text Available Abstract The draft genome of Pectobacterium carotovorum subsp. brasiliense (Pcb which causes blackleg of potato was submitted to the NCBI and released with reference number NZ_LGRF00000000.1. The estimated genome size based on the draft genome assembly is 4,820,279 bp from 33 contigs ranging in length from 444 to 1,660,019 nucleotides. The genome annotation showed 4250 putative genes, 4114 CDS and 43 pseudo-genes. Three complete rRNA gene species were detected: nine 5S, one 16S and one 23S. Other partial rRNA gene fragments were also identified, nine 16S rRNA and three 23S rRNA. A total of 69 tRNA genes and one ncRNA gene were also annotated in this genome.

  17. [Dissociation of Pectobacterium carotovorum subsp. carotovorumrelated to the changes in the cell wall lipopolysaccharide].

    Science.gov (United States)

    Tovkach, F I; Romaniuk, L V; Gorb, T E; Ostapchuk, A N; Muchnik, F V

    2012-01-01

    It is shown for the first time that population heterogeneity of Pectobacterium carotovorum subsp. carotovorum is applicable to a wide range of strains and therefore is a universal characteristic. Using the method of specific selection with the help of carotovoricins which are identical to the phage tails a set of population dissociants of different types was obtained, due to the fact that S-LPS is the part of the cell wall which contains their attachment sites. It was determined that changes in S-lipopolysaccharides lead to the formation of SR-, R-forms of P. carotovorum. A suggestion is made that the changes in the surface structures of dissociants have a significant impact on secretion types II and III--the main pathogenicity factor of some bacteria. The results presented are a prerequisite for studying the direction, the reasons for dissociation process and its impact on the pathogenicity of P. carotovorum.

  18. Germination and conjugation of Bacillus thuringiensis subsp. israelensis in the intestine of gnotobiotic rats

    DEFF Research Database (Denmark)

    Wilcks, Andrea; Ørum-Smidt, Lasse; Bahl, Martin Iain

    2008-01-01

    Aims: To study the ability of Bacillus thuringiensis subsp. israelensis spores to germinate and subsequently transfer a conjugative plasmid in the intestinal tract of gnotobiotic rats. Methods and Results: Germination was studied by feeding germ-free rats with spores of a B. thuringiensis strain...... harbouring a plasmid encoding green fluorescent protein (GFP), which enabled quantification of germinated bacteria by flow cytometry. To study in vivo conjugation, germ-free rats were first associated with a B. thuringiensis recipient strain and after 1 week an isogenic donor strain harbouring...... the conjugative plasmid pXO16 was introduced. Both strains were given as spores and transfer of pXO16 was observed from the donor to the recipient strain. Conclusions: Bacillus thuringiensis is able to have a full life cycle in the intestine of gnotobiotic rats including germination of spores, several cycles...

  19. Mechanism of inhibited growth of Bacillus pumilus by Propionibacterium freudenreichii subsp. shermanii.

    Science.gov (United States)

    Marshall, D L; Odame-Darkwah, J K

    1994-04-01

    Physiological studies were conducted in an attempt to elucidate the mechanism of inhibition of Bacillus pumilus by Propionibacterium freudenreichii subsp. shermanii. Inhibition of B. pumilus by P. shermanii occurred in media supplemented with 1% glucose, indicating that glucose utilization by the latter bacterium was not responsible for growth inhibition of the former bacterium. The medium pH in which P. shermanii inhibited the growth of B. pumilus was 4.3. Propionic acid was positively identified in the culture medium in which B. pumilus was inhibited by P. shermanii. The presence of propionic acid and a low medium pH may account for the inhibition of B. pumilus by P. shermanii. Sodium lactate concentrations of 0.8-1.0% were essential for the continuous growth of and propionic acid production by P. shermanii. Thus, use of P. shermanii to inhibit B. pumilus in foods would likely require a lactate source.

  20. Morphological and ultrastructural studies on Ulva flexuosa subsp. pilifera (Chlorophyta from Poland

    Directory of Open Access Journals (Sweden)

    Beata Messyasz

    2013-06-01

    Full Text Available Ulva flexuosa subsp. pilifera (Kütz. M. J. Wynne 2005 (= Enteromorpha pilifera Kützing 1845 was previously found in Argentina, the Czech Republic, Germany, Hungary, Romania, Slovakia and Sweden, recently also in Poland. The genus Ulva was first time described as Enteromorpha. Interestingly, Enteromorpha is used nowadays as a synonym for Ulva, a development which is based on molecular data. The morphologies of both young and mature specimens were studied, and most life cycle stages could be observed. Further, the formation of calcium carbonate crystals on the surface of Ulva thalli seems to influence the arrangement of the cells. A detailed ultrastructural (TEM analysis of cell walls is presented. The TEM reveals in great details highly complex, irregular structures with stratification lines.

  1. Chemical Composition Variability of the Herb Essential Oil in the Ontogenesis of Artemisia campestris subsp. campestris.

    Science.gov (United States)

    Lis, Anna; Kowal, Martyna; Kończak, Joanna

    2015-10-01

    The essential oil from the herb of Artemisia campestris L. subsp. campestris harvested at five ontogenesis phases was analyzed by GC, GC/MS and 1H NMR spectroscopy. More than 130 compounds were identified. Terpene hydrocarbons dominated in the oil. Germacrene D (20.3-30.1%), β-pinene (3.7-15.4%) and γ-humulene (6.6-9.8%) were the main constituents. The contents of germacrene D and γ-humulene decreased from the phase before bud formation to the beginning of flowering, then increased until the post-flowering phase. The content of β-pinene at the same periods firstly increased, and then was reduced.

  2. Antimicrobial resistance in Salmonella enterica subsp. enterica serovar typhimurium from humans and production animals

    DEFF Research Database (Denmark)

    Seyfarth, Anne Mette; Wegener, Henrik Caspar; FrimodtMoller, N.

    1997-01-01

    .4% of strains from cattle, 11.1% of strains from pigs and 9.2% of strains from poultry. Multiple resistance, i.e. resistance against at least four antimicrobial agents, was found in 9.2% of the human strains, but in only two of the cattle isolates, The majority of the multi-resistant strains in humans were from......: Poultry strains were usually resistant only to ampicillin, white pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance......We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp, enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent...

  3. Apparent Prevalence of Beef Carcasses Contaminated with Mycobacterium avium subsp. paratuberculosis Sampled from Danish Slaughter Cattle

    Directory of Open Access Journals (Sweden)

    Hisako Okura

    2011-01-01

    Full Text Available Presence of Mycobacterium avium subsp. paratuberculosis (MAP in beef has been reported as a public health concern because asymptomatically infected cattle may contain MAP in tissues that are used for human consumption. Associations between MAP carcasses contamination and animal characteristics such as age, breed, production type, and carcass classification were assessed. Cheek muscles from 501 carcasses were sampled cross-sectionally at a Danish abattoir and tested for presence of viable MAP and MAP DNA by bacterial culture and IS900 realtime PCR, respectively. Cheek muscle tissues from carcasses of two dairy cows were positive by culture whereas 4% of the animals were estimated with ≥10 CFU/gram muscle based on realtime PCR. Age was found to be associated with carcass contamination with MAP. The observed viable MAP prevalence in beef carcasses was low. However, detection of MAP and MAP DNA in muscle tissues suggested that bacteremia occurred in slaughtered cattle.

  4. Antimicrobial resistance in Salmonella enterica subsp. enterica serovar typhimurium from humans and production animals

    DEFF Research Database (Denmark)

    Seyfarth, Anne Mette; Wegener, Henrik Caspar; FrimodtMoller, N.

    1997-01-01

    infections contracted outside Denmark, most often in southern Europe or south-east Asia. Resistance in human strains was most common against tetracycline (13%), ampicillin (12%), sulphonamide (12%), streptomycin (10%) and chloramphenicol (8%). The resistance pattern differed somewhat in animal isolates......: Poultry strains were usually resistant only to ampicillin, white pig and cattle isolates were most often resistant to sulphonamide, tetracycline and streptomycin. Typing of the strains showed that some animal strains and human strains were indistinguishable. In conclusion, while antimicrobial resistance......We have studied the frequency of antimicrobial resistance and epidemiological relatedness among 473 isolates of Salmonella enterica subsp, enterica serovar typhimurium (S. typhimurium) from human and veterinary sources. The human strains were clinical isolates from patients with diarrhoea sent...

  5. Effect of Weather on the Occurrence of Puccinia Graminis Subsp. Graminicola and Puccinia Coronata F. Sp. Lolii at Lolium Perenne L. and Deschampsia Caespitosa (L. P. B.

    Directory of Open Access Journals (Sweden)

    Monika Novotná

    2017-01-01

    Full Text Available Monitoring of Puccinia graminis subsp. graminicola and Puccinia coronata f. sp. lolii was carried out in Plant breeding station called Větrov. The pathogens were estimated on turf grass (Lolium perenne L., Deschampsia caespitosa (L. P. B. from 2009 to 2014. Puccinia graminis subsp. graminicola was detected in the increased level in 2009 and 2012. The highest amount of mixed infections was determined in 2014 because of the warmest winter from all monitored years and low precipitations. Significant differences were found out in the resistance of similar plant materials grown in different fields. Significant effect of weather conditions and supposed effect of different infectious pressure on various fields were reflected in these facts. At evaluated grasses, the highest (P < 0.05 occurence of Puccinia graminis subsp. graminicola. Lolium perenne L. was observed and the infection of Puccinia graminis subsp. graminicola (P < 0.05 was determined higher than in Deschampsia caespitosa (L. P. B.

  6. Prevention of hypertension-induced vascular dementia by Lactobacillus paracasei subsp. paracasei NTU 101-fermented products.

    Science.gov (United States)

    Cheng, Meng-Chun; Pan, Tzu-Ming

    2017-12-01

    Numerous etiological studies have established positive clinical association between hypertension and vascular dementia (VaD). Lactobacillus paracasei subsp. paracasei NTU 101-fermented products have been shown to decrease vascular risk factors such as hypertension, atherosclerosis, hyperlipidemia and obesity. This study investigated the effect of ethanol extract of Lactobacillus paracasei subsp. paracasei NTU 101-fermented products (NTU101F) in hypertension-induced VaD in rats. Hypertension was promoted by subcutaneous injection of deoxycorticosterone acetate (DOCA, 25 mg/kg body weight/day, twice a week) and substitution of drinking water with 1.0% NaCl and 0.2% KCl. The NTU101F groups (0.5, 1.0, and 5.0) administered NTU101F at the concentrations 11, 22, and 110 mg/kg body weight/day, respectively, starting from day 51 day of DOCA-salt treatment. Morris water maze (MWM) was used for testing learning and memory. Different biochemical estimations were used to assess oxidative stress and inflammatory response in hippocampus. Oral administration of NTU101F in DOCA-salt hypertension-induced VaD rats resulted in a significant decrease in blood pressure by 18.3-23.2% (p < 0.001), which was regulated by increasing eNOS density (about 3-fold) in the aorta, promoting NO production, and decreasing of matrix metallopeptidase 9 activity (about 2-fold) in the hippocampus, in addition to improve the kidney function and structure, decrease escape latency and increase the times spent in the target quadrant by 23.5-27.8% (p < 0.05). Overall, our findings suggest that NTU101F could exert neuroprotection in the brain and attenuate hypertension-induced VaD.

  7. Production of the bioinsecticide Bacillus thuringiensis subsp. israelensis with deltamethrin increases toxicity towards mosquito larvae.

    Science.gov (United States)

    Tetreau, G; Patil, C D; Chandor-Proust, A; Salunke, B K; Patil, S V; Després, L

    2013-08-01

    Bacillus thuringiensis subsp. israelensis is a bioinsecticide used for larval mosquito control and it represents a safe alternative to chemical insecticides. Despite its environmental safety, it is less efficient and persistent than chemical insecticides. To bypass these limitations, we propose to combine the advantages of chemical and biological insecticides by producing Bti in a medium supplemented with a chemical insecticide (DDT, deltamethrin, permethrin, propoxur or temephos). Among the investigated insecticides, the addition of deltamethrin in the medium induced a higher toxicity (over 6.72-fold) of the composite deltamethrin-Bti towards mosquito larvae as compared to Bti alone. This was mainly due to the insertion of deltamethrin into the membranes of Bti spores, as evidenced by a quantification of membrane-extracted deltamethrin by HPLC. This composite larvicide is a promising tool to decrease the quantity of chemicals dispersed in the environment, to increase the efficacy of Bti and to facilitate its widespread use as a transition between chemical and biological insecticides. Further experiments are required to characterize the mechanisms that underline the incorporation of deltamethrin into Bti to optimize the production and the toxicity of this composite larvicide. This study is the first report of an increased efficacy of the mosquitocidal bioinsecticide Bacillus thuringiensis subsp. israelensis (Bti) when produced with a chemical insecticide. The results clearly demonstrate that deltamethrin is able to synergize the insecticidal activity of Bti through inclusion into spore membranes, reducing off-target and nonspecific toxicity occurring when the chemical is used alone as sprays. This new composite chemical-biological insecticide can become an invaluable tool as an intermediate between single chemical usage and the widespread use of Bti, notably in developing countries with limited financial resources for intensive mosquito control campaigns. © 2013

  8. Identification and Characterization of Three Previously Undescribed Crystal Proteins from Bacillus thuringiensis subsp. jegathesan

    Science.gov (United States)

    Sun, Yunjun; Zhao, Qiang; Ding, Xuezhi; Hu, Quanfang; Federici, Brian A.

    2013-01-01

    The total protoxin complement in the parasporal body of mosquitocidal strain, Bacillus thuringiensis subsp. jegathesan 367, was determined by use of a polyacrylamide gel block coupled to mass spectrometry. A total of eight protoxins were identified from this strain, including five reported protoxins (Cry11Ba, Cry19Aa, Cry24Aa, Cry25Aa, and Cyt2Bb), as well as three previously undescribed (Cry30Ca, Cry60Aa, and Cry60Ba) in this isolate. It was interesting that the encoding genes of three new protoxins existed as cry30Ca-gap-orf2 and cry60Ba-gap-cry60Aa. The cry30Ca and a downstream orf2 gene were oriented in the same direction and separated by 114 bp, and cry60Ba was located 156 bp upstream from and in the same orientation to cry60Aa. The three new protoxin genes were cloned from B. thuringiensis subsp. jegathesan and expressed in an acrystalliferous strain under the control of cyt1A gene promoters and the STAB-SD stabilizer sequence. Recombinant strain containing only cry30Ca did not produce visible inclusion under microscope observation, while that containing both cry30Ca and orf2 could produce large inclusions. Cry60Aa and Cry60Ba synthesized either alone or together in the acrystalliferous host could yield large inclusions. In bioassays using the fourth-instar larvae of Culex quinquefasciatus, Cry60Aa and Cry60Ba alone or together had estimated 50% lethal concentrations of 2.9 to 7.9 μg/ml; however, Cry30Ca with or without ORF2 was not toxic to this mosquito. PMID:23524673

  9. Long lasting persistence of Bacillus thuringiensis Subsp. israelensis (Bti in mosquito natural habitats.

    Directory of Open Access Journals (Sweden)

    Mathieu Tilquin

    Full Text Available BACKGROUND: The detrimental effects of chemical insecticides on the environment and human health have lead to the call for biological alternatives. Today, one of the most promising solutions is the use of spray formulations based on Bacillus thuringiensis subsp. israelensis (Bti in insect control programs. As a result, the amounts of Bti spread in the environment are expected to increase worldwide, whilst the common belief that commercial Bti is easily cleared from the ecosystem has not yet been clearly established. METHODOLOGY/MAIN FINDINGS: In this study, we aimed to determine the nature and origin of the high toxicity toward mosquito larvae found in decaying leaf litter collected in several natural mosquito breeding sites in the Rhône-Alpes region. From the toxic fraction of the leaf litter, we isolated B. cereus-like bacteria that were further characterized as B. thuringiensis subsp. israelensis using PCR amplification of specific toxin genes. Immunological analysis of these Bti strains showed that they belong to the H14 group. We finally used amplified length polymorphism (AFLP markers to show that the strains isolated from the leaf litter were closely related to those present in the commercial insecticide used for field application, and differed from natural worldwide genotypes. CONCLUSIONS/SIGNIFICANCE: Our results raise the issue of the persistence, potential proliferation and environmental accumulation of human-spread Bti in natural mosquito habitats. Such Bti environmental persistence may lengthen the exposure time of insects to this bio-insecticide, thereby increasing the risk of resistance acquisition in target insects, and of a negative impact on non-target insects.

  10. Characterization of nitrite degradation by Lactobacillus casei subsp. rhamnosus LCR 6013.

    Directory of Open Access Journals (Sweden)

    Dong-mei Liu

    Full Text Available Nitrites are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. The nitrite degradation capacity of Lactobacillus casei subsp. rhamnosus LCR 6013 was investigated in pickle fermentation. After LCR 6013 fermentation for 120 h at 37°C, the nitrite concentration in the fermentation system was significantly lower than that in the control sample without the LCR 6013 strain. The effects of NaCl and Vc on nitrite degradation by LCR 6013 in the De Man, Rogosa and Sharpe (MRS medium were also investigated. The highest nitrite degradations, 9.29 mg/L and 9.89 mg/L, were observed when NaCl and Vc concentrations were 0.75% and 0.02%, respectively in the MRS medium, which was significantly higher than the control group (p ≤ 0.01. Electron capture/gas chromatography and indophenol blue staining were used to study the nitrite degradation pathway of LCR 6013. The nitrite degradation products contained N2O, but no NH4(+. The LCR 6013 strain completely degraded all NaNO2 (50.00 mg/L after 16 h of fermentation. The enzyme activity of NiR in the periplasmic space was 2.5 times of that in the cytoplasm. Our results demonstrated that L. casei subsp. rhamnosus LCR 6013 can effectively degrade nitrites in both the pickle fermentation system and in MRS medium by NiR. Nitrites are degraded by the LCR 6013 strain, likely via the nitrate respiration pathway (NO2(->NO->N2O->N2, rather than the aammonium formation pathway (dissimilatory nitrate reduction to ammonium, DNRA, because the degradation products contain N2O, but not NH4(+.

  11. Transcriptome profiling of Lactococcus lactis subsp. cremoris CECT 8666 in response to agmatine

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    Beatriz del Rio

    2016-03-01

    Full Text Available The dairy strain Lactococcus lactis subsp. cremoris CECT 8666 (formerly GE2-14 synthesizes the biogenic amine putrescine from agmatine via the agmatine deiminase (AGDI pathway [1]. The AGDI cluster of L. lactis is composed by five genes aguR, aguB, aguD, aguA and aguC. The last four genes are co-transcribed as a single policistronic mRNA forming the catabolic operon aguBDAC, which encodes the proteins necessary for agmatine uptake and its conversion into putrescine [1,2]. The first gene of the cluster, aguR, encodes a transmembrane protein that functions as a one-component signal transduction system that senses the agmatine concentration of the medium and accordingly regulates the transcription of aguBDAC [2]. The catabolic operon aguBDAC is transcriptionally activated by agmatine [2] and transcriptionally regulated by carbon catabolite repression (CCR via glucose, but not by other sugars such as lactose or galactose [1,3]. On the contrary, the transcription of the aguR regulatory gene is not subject to CCR regulation [1,3] nor is regulated by agmatine [2]. In this study we report the transcriptional profiling of L. lactis subsp. cremoris CECT 8666 grown in M17 medium with galactose (GalM17 as carbon source and supplemented with agmatine, compared to that of the strain grown in the same culture medium without agmatine. The transcriptional profiling data of agmatine-regulated genes were deposited in the Gene Expression Omnibus (GEO database under Accession no. GSE74808.

  12. The molecular identification of Streptococcus equi subsp. equi strains isolated within New Zealand.

    Science.gov (United States)

    Patty, O A; Cursons, R T M

    2014-03-01

    To identify Streptococcus equi subsp. equi (S. equi) by PCR analysis and obtain isolates by culture, in order to investigate the strains of S. equi infecting horses within New Zealand. A diagnostic PCR, based on the amplification of the seeI gene for S. equi, was used on 168 samples submitted from horses with and without clinical signs of strangles. Samples were also processed and cultured on selective media for the isolation of β-haemolytic colonies. In addition, the hypervariable region of the seM gene of S. equi was amplified and then sequenced for strain typing purposes. Of the 168 samples, 35 tested positive for S. equi using PCR. Thirty-two confirmed samples were from horses with a clinical diagnosis of strangles and three were from horses where clinical information was unavailable. Only 22/35 (63%) confirmed S. equi samples were successfully isolated following culture. Strain typing demonstrated that two novel seM alleles of S. equi were found in New Zealand with SeM-99 strains being restricted to the North Island while SeM-100 strains were found in both North and South Islands. The application of PCR for the laboratory confirmation of strangles allowed for a rapid and sensitive identification of S. equi. Moreover, seM typing revealed that within the samples examined two strains of S. equi co-circulated within the North Island of New Zealand but only one strain in the South Island. PCR reduces the time required to obtain laboratory confirmation of strangles compared with culture methods. It also has greater sensitivity in detecting S. equi infections, which is of particular importance in the detection of carrier animals which normally shed low numbers of bacteria. Additionally, seM molecular typing can differentiate between bacterial strains, assisting in the monitoring of local strains of S. equi subsp. equi causing disease.

  13. Characterization of novel microsatellite markers in Musa acuminata subsp. burmannicoides, var. Calcutta 4

    Directory of Open Access Journals (Sweden)

    Amorim Edson P

    2010-05-01

    Full Text Available Abstract Background Banana is a nutritionally important crop across tropical and sub-tropical countries in sub-Saharan Africa, Central and South America and Asia. Although cultivars have evolved from diploid, triploid and tetraploid wild Asian species of Musa acuminata (A genome and Musa balbisiana (B genome, many of today's commercial cultivars are sterile triploids or diploids, with fruit developing via parthenocarpy. As a result of restricted genetic variation, improvement has been limited, resulting in a crop frequently lacking resistance to pests and disease. Considering the importance of molecular tools to facilitate development of disease resistant genotypes, the objectives of this study were to develop polymorphic microsatellite markers from BAC clone sequences for M. acuminata subsp. burmannicoides, var. Calcutta 4. This wild diploid species is used as a donor cultivar in breeding programs as a source of resistance to diverse biotic stresses. Findings Microsatellite sequences were identified from five Calcutta 4 BAC consensi datasets. Specific primers were designed for 41 loci. Isolated di-nucleotide repeat motifs were the most abundant, followed by tri-nucleotides. From 33 tested loci, 20 displayed polymorphism when screened across 21 diploid M. acuminata accessions, contrasting in resistance to Sigatoka diseases. The number of alleles per SSR locus ranged from two to four, with a total of 56. Six repeat classes were identified, with di-nucleotides the most abundant. Expected heterozygosity values for polymorphic markers ranged from 0.31 to 0.75. Conclusions This is the first report identifying polymorphic microsatellite markers from M. acuminata subsp. burmannicoides, var. Calcutta 4 across accessions contrasting in resistance to Sigatoka diseases. These BAC-derived polymorphic microsatellite markers are a useful resource for banana, applicable for genetic map development, germplasm characterization, evolutionary studies and marker

  14. Characterization of novel microsatellite markers in Musa acuminata subsp. burmannicoides, var. Calcutta 4.

    Science.gov (United States)

    Miller, Robert Ng; Passos, Marco An; Menezes, Natalia Np; Souza, Manoel T; do Carmo Costa, Marcos M; Rennó Azevedo, Vânia C; Amorim, Edson P; Pappas, Georgios J; Ciampi, Ana Y

    2010-05-27

    Banana is a nutritionally important crop across tropical and sub-tropical countries in sub-Saharan Africa, Central and South America and Asia. Although cultivars have evolved from diploid, triploid and tetraploid wild Asian species of Musa acuminata (A genome) and Musa balbisiana (B genome), many of today's commercial cultivars are sterile triploids or diploids, with fruit developing via parthenocarpy. As a result of restricted genetic variation, improvement has been limited, resulting in a crop frequently lacking resistance to pests and disease. Considering the importance of molecular tools to facilitate development of disease resistant genotypes, the objectives of this study were to develop polymorphic microsatellite markers from BAC clone sequences for M. acuminata subsp. burmannicoides, var. Calcutta 4. This wild diploid species is used as a donor cultivar in breeding programs as a source of resistance to diverse biotic stresses. Microsatellite sequences were identified from five Calcutta 4 BAC consensi datasets. Specific primers were designed for 41 loci. Isolated di-nucleotide repeat motifs were the most abundant, followed by tri-nucleotides. From 33 tested loci, 20 displayed polymorphism when screened across 21 diploid M. acuminata accessions, contrasting in resistance to Sigatoka diseases. The number of alleles per SSR locus ranged from two to four, with a total of 56. Six repeat classes were identified, with di-nucleotides the most abundant. Expected heterozygosity values for polymorphic markers ranged from 0.31 to 0.75. This is the first report identifying polymorphic microsatellite markers from M. acuminata subsp. burmannicoides, var. Calcutta 4 across accessions contrasting in resistance to Sigatoka diseases. These BAC-derived polymorphic microsatellite markers are a useful resource for banana, applicable for genetic map development, germplasm characterization, evolutionary studies and marker assisted selection for traits.

  15. Prévalence de Acidovorax avenae subsp. avenae, agent des rayures bactériennes du riz dans les semences de base produites au Burkina Faso

    OpenAIRE

    Dakouo, D.; Ouedraogo, SL.; Somda, I.; Mortensen, CN.

    2005-01-01

    Prevalence of Bacterial Stripe Organism, Acidovorax avenae subsp. avenae, in Breeder Rice Seed Samples from Burkina Faso. Nine rice seed samples of improved and local varieties were tested at DGISP (Denmark) for the incidence of seed-borne bacterial stripe organism, Acidovorax avenae subsp. avenae, using the cassette holder method. Twenty-six suspected bacterial colonies were identified by different methods including colony morphology, pigmentation, biochemical and pathogenicity tests. Using ...

  16. A randomised double-blind placebo-controlled trial with Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 for maintenance of remission in ulcerative colitis

    DEFF Research Database (Denmark)

    Wildt, Signe; Nordgaard, Inge; Hansen, Ulla

    2011-01-01

    To investigate the clinical effect of treatment with Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 (Probio-Tec AB-25) to maintain remission in patients with ulcerative colitis.......To investigate the clinical effect of treatment with Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 (Probio-Tec AB-25) to maintain remission in patients with ulcerative colitis....

  17. Some biological compounds, radical scavenging capacities and antimicrobial activities in the seeds of Nepeta italica L. and Sideritis montana L. subsp. montana from Turkey

    Energy Technology Data Exchange (ETDEWEB)

    Emre, I.; Kursat, M.; Yilmaz, O.; Erecevit, P.

    2011-07-01

    This study determined some biological compounds (fatty acid compositions, lipid-soluble vitamins, sterols, flavonoids), radical scavenging capacities and antimicrobial activities in the seeds of Nepeta italica L. and Sideritis montana L. subsp. montana. It was found that palmitic acid (C16:0; 8.54+-0.13-3.05+-0.04%), oleic acid (C18:1 n9, 22.41+-0.8-18.83+-0.1%) and a-inolenic acid were the dominant fatty acids in both Nepeta italica L. and Sideritis montana L. subsp. montana. It was concluded that both Nepeta italica L. and Sideritis montana L. subsp. montana contained stigmasterol and ergosterol as well as beta-sitosterol. The present findings show that Nepeta italica L. contains morin, catechin, naringin and Sideritis montana L. subsp. montana contains morin, naringenin as major flavonoids. It was also determined that methanol extracts of Nepeta italica L. and Sideritis montana L. subsp. montana were most effective against DPPH radicals. The results of the present study show that the vitamins, flavonoids and fatty acid extracts in the seeds of N. italica L. and S. montana L. subsp. montana prevented the growth of the microorganisms used in the tests at different ratios. (Author).

  18. A quantitative and direct PCR assay for the subspecies-specific detection of Clavibacter michiganensis subsp. michiganensis based on a ferredoxin reductase gene.

    Science.gov (United States)

    Cho, Min Seok; Lee, Jang Ha; Her, Nam Han; Kim, Changkug; Seol, Young-Joo; Hahn, Jang Ho; Baeg, Ji Hyoun; Kim, Hong Gi; Park, Dong Suk

    2012-06-01

    The Gram-positive bacterium Clavibacter michiganensis subsp. michiganensis is the causal agent of canker disease in tomato. Because it is very important to control newly introduced inoculum sources from commercial materials, the specific detection of this pathogen in seeds and seedlings is essential for effective disease control. In this study, a novel and efficient assay for the detection and quantitation of C. michiganensis subsp. michiganensis in symptomless tomato and red pepper seeds was developed. A pair of polymerase chain reaction (PCR) primers (Cmm141F/R) was designed to amplify a specific 141 bp fragment on the basis of a ferredoxin reductase gene of C. michiganensis subsp. michiganensis NCPPB 382. The specificity of the primer set was evaluated using purified DNA from 16 isolates of five C. michiganensis subspecies, one other Clavibacter species, and 17 other reference bacteria. The primer set amplified a single band of expected size from the genomic DNA obtained from the C. michiganensis subsp. michiganensis strains but not from the other C. michiganensis subspecies or from other Clavibacter species. The detection limit was a single cloned copy of the ferredoxin reductase gene of C. michiganensis subsp. michiganensis. In conclusion, this quantitative direct PCR assay can be applied as a practical diagnostic method for epidemiological research and the sanitary management of seeds and seedlings with a low level or latent infection of C. michiganensis subsp. michiganensis.

  19. Evaluation of the effects of Olea europaea L. subsp. africana (Mill.) P.S. Green (Oleaceae) leaf methanol extract against castor oil-induced diarrhoea in mice.

    Science.gov (United States)

    Amabeoku, George J; Bamuamba, Kapinga

    2010-03-01

    Olea europaea L. subsp. africana (Mill.) P.S. Green is widely used in South Africa by traditional medicine practitioners to treat diarrhoea. However, little is known scientifically about this South African species in the treatment of diarrhoea. The main aim of the study therefore was to investigate the antidiarrhoeal effect of the leaf methanol extract of the plant species in mice. The antidiarrhoeal activity of the leaf methanol extract of O. europaea subsp. africana was studied using a castor oil-induced diarrhoeal test. The antipropulsive activity of the plant extract was also investigated using the charcoal meal transit test. Standard methods were used to investigate the acute toxicity and effect of O. europaea subsp. africana on castor oil-induced intraluminal fluid accumulation. Leaf methanol extract of O. europaea subsp. africana and loperamide, a standard antidiarrhoeal drug, significantly reduced the number of diarrhoeal episodes induced by castor oil, significantly decreased the stool mass, significantly delayed the onset of the diarrhoea and protected the animals against castor oil-induced diarrhoea. Both O. europaea subsp. africana and loperamide significantly decreased the gastrointestinal transit of charcoal meal and castor oil-induced intraluminal fluid accumulation in mice. The LD50 value was found to be 3475 mg/kg (p.o.). The results obtained suggest that the leaf methanol extract of O. europaea subsp. africana has an antidiarrhoeal property and that, given orally, it may be non-toxic and/or safe in mice.

  20. Prevalence and comparison of Streptococcus infantarius subsp. infantarius and Streptococcus gallolyticus subsp. macedonicus in raw and fermented dairy products from East and West Africa.

    Science.gov (United States)

    Jans, Christoph; Kaindi, Dasel Wambua Mulwa; Böck, Désirée; Njage, Patrick Murigu Kamau; Kouamé-Sina, Sylvie Mireille; Bonfoh, Bassirou; Lacroix, Christophe; Meile, Leo

    2013-10-15

    Streptococcus infantarius subsp. infantarius (Sii) and Streptococcus gallolyticus subsp. macedonicus are members of the Streptococcus bovis/Streptococcus equinus complex (SBSEC) associated with human infections. SBSEC-related endocarditis was furthermore associated with rural residency in Southern Europe. SBSEC members are increasingly isolated as predominant species from fermented dairy products in Europe, Asia and Africa. African variants of Sii displayed dairy adaptations to lactose metabolism paralleling those of Streptococcus thermophilus including genome decay. In this study, the aim was to assess the prevalence of Sii and possibly other SBSEC members in dairy products of East and West Africa in order to identify their habitat, estimate their importance in dairy fermentation processes and determine geographic areas affected by this potential health risk. Presumptive SBSEC members were isolated on semi-selective M17 and SM agar media. Subsequent genotypic identification of isolates was based on rep-PCR fingerprinting and SBSEC-specific16S rRNA gene PCR assay. Detailed identification was achieved through application of novel primers enhancing the binding stringency in partial groES/groEL gene amplification and subsequent DNA sequencing. The presence of S. thermophilus-like lacS and lacZ genes in the SBSEC isolates was determined to elucidate the prevalence of this dairy adaptation. Isolates (n = 754) were obtained from 72 raw and 95 fermented milk samples from Côte d'Ivoire and Kenya on semi-selective agar media. Colonies of Sii were not detected from raw milk despite high microbial titers of approximately 10(6)CFU/mL on M17 agar medium. However, after spontaneous milk fermentation Sii was genotypically identified in 94.1% of Kenyan samples and 60.8% of Kenyan isolates. Sii prevalence in Côte d'Ivoire displayed seasonal variations in samples from 32.3% (June) to 40.0% (Dec/Jan) and isolates from 20.5% (June) to 27.7% (Dec/Jan) present at titers of 10

  1. Identification of genomic differences between Campylobacter jejuni subsp. jejuni and C. jejuni subsp. doylei at the nap locus leads to the development of a C. jejuni subspeciation multiplex PCR method

    Directory of Open Access Journals (Sweden)

    Heath Sekou

    2007-02-01

    Full Text Available Abstract Background The human bacterial pathogen Campylobacter jejuni contains two subspecies: C. jejuni subsp. jejuni (Cjj and C. jejuni subsp. doylei (Cjd. Although Cjd strains are isolated infrequently in many parts of the world, they are obtained primarily from human clinical samples and result in an unusual clinical symptomatology in that, in addition to gastroenteritis, they are associated often with bacteremia. In this study, we describe a novel multiplex PCR method, based on the nitrate reductase (nap locus, that can be used to unambiguously subspeciate C. jejuni isolates. Results Internal and flanking napA and napB primer sets were designed, based on existing C. jejuni and Campylobacter coli genome sequences to create two multiplex PCR primer sets, nap mpx1 and nap mpx2. Genomic DNA from 161 C. jejuni subsp. jejuni (Cjj and 27 C. jejuni subsp. doylei (Cjd strains were amplified with these multiplex primer sets. The Cjd strains could be distinguished clearly from the Cjj strains using either nap mpx1 or mpx2. In addition, combination of either nap multiplex method with an existing lpxA speciation multiplex method resulted in the unambiguous and simultaneous speciation and subspeciation of the thermophilic Campylobacters. The Cjd nap amplicons were also sequenced: all Cjd strains tested contained identical 2761 bp deletions in napA and several Cjd strains contained deletions in napB. Conclusion The nap multiplex PCR primer sets are robust and give a 100% discrimination of C. jejuni subspecies. The ability to rapidly subspeciate C. jejuni as well as speciate thermophilic Campylobacter species, most of which are pathogenic in humans, in a single amplification will be of value to clinical laboratories in strain identification and the determination of the environmental source of campylobacterioses caused by Cjd. Finally, the sequences of the Cjd napA and napB loci suggest that Cjd strains arose from a common ancestor, providing clues as to

  2. Isolation of Bartonella henselae, Bartonella koehlerae subsp. koehlerae, Bartonella koehlerae subsp. bothieri and a new subspecies of B. koehlerae from free-ranging lions (Panthera leo) from South Africa, cheetahs (Acinonyx jubatus) from Namibia and captive cheetahs from California.

    Science.gov (United States)

    Molia, S; Kasten, R W; Stuckey, M J; Boulouis, H J; Allen, J; Borgo, G M; Koehler, J E; Chang, C C; Chomel, B B

    2016-11-01

    Bartonellae are blood- and vector-borne Gram-negative bacteria, recognized as emerging pathogens. Whole-blood samples were collected from 58 free-ranging lions (Panthera leo) in South Africa and 17 cheetahs (Acinonyx jubatus) from Namibia. Blood samples were also collected from 11 cheetahs (more than once for some of them) at the San Diego Wildlife Safari Park. Bacteria were isolated from the blood of three (5%) lions, one (6%) Namibian cheetah and eight (73%) cheetahs from California. The lion Bartonella isolates were identified as B. henselae (two isolates) and B. koehlerae subsp. koehlerae. The Namibian cheetah strain was close but distinct from isolates from North American wild felids and clustered between B. henselae and B. koehlerae. It should be considered as a new subspecies of B. koehlerae. All the Californian semi-captive cheetah isolates were different from B. henselae or B. koehlerae subsp. koehlerae and from the Namibian cheetah isolate. They were also distinct from the strains isolated from Californian mountain lions (Felis concolor) and clustered with strains of B. koehlerae subsp. bothieri isolated from free-ranging bobcats (Lynx rufus) in California. Therefore, it is likely that these captive cheetahs became infected by an indigenous strain for which bobcats are the natural reservoir.

  3. Comparison of the acidifying activity of Lactococcus lactis subsp. lactis strains isolated from goat's milk and Valdeteja cheese.

    Science.gov (United States)

    Alonso-Calleja, C; Carballo, J; Capita, R; Bernardo, A; García-López, M L

    2002-01-01

    This work was carried out to study the acid production by Lactococcus lactis subsp. lactis strains isolated from goat's milk and goat cheese (Valdeteja variety) in order to select a suitable starter culture for industrial goat cheese manufacturing. The titrable acidity of 45 Lactococcus lactis subsp. lactis strains isolated from a home-made batch of Valdeteja cheese with excellent sensory characteristics was measured over a period of 18 h. The strains were divided into two groups depending on the acid production rate: 20 fast acid producer (F) strains and 25 slow acid producer (S) strains. The kinetic parameters (lag phase, maximum acid production rate and value of upper asymptote curve) of the acid production curves for F and S strains were significantly (P cheese manufacturing.

  4. Transcriptional responses in Lactococcus lactis subsp. cremoris to the changes in oxygen and redox potential during milk acidification

    DEFF Research Database (Denmark)

    Larsen, Nadja; Werner, Birgit Brøsted; Jespersen, Lene

    2016-01-01

    is important for the better control and reproducibility of dairy fermentations. We applied mRNA sequencing by Illumina HiSeq 2000 to investigate gene expression profile in a dairy strain of Lactococcus lactis subsp. cremoris during milk acidification. Novelty of this study lies in linking transcriptional...... responses to oxygen depletion and the changes of redox potential with the fermentation kinetics and clarification of molecular factors specifically expressed in milk which might be essential for bacterial performance and the final quality of cheeses.......Milk acidification and metabolic activity of the starter cultures are affected by oxygen; however, molecular factors related to the redox changes are poorly defined. The objective of the study was to investigate transcriptional responses in Lactococcus lactis subsp. cremoris CHCCO2 grown in milk...

  5. Quorum sensing in Aeromonas salmonicida subsp. achromogenes and the effect of the autoinducer synthase AsaI on bacterial virulence

    DEFF Research Database (Denmark)

    Schwenteit, Johanna; Gram, Lone; Nielsen, Kristian Fog

    2011-01-01

    The Gram-negative fish pathogenic bacterium Aeromonas salmonicida possesses the LuxIRtype quorum sensing (QS) system, termed AsaIR. In this study the role of QS in A. salmonicida subsp. achromogenes virulence and pigment production was investigated. Five wild-type Asa strains induced the N......Ideficient mutant was 20-fold higher than that of the isogenic wt strain and the mean day to death of the mutant was significantly prolonged. Furthermore, the expression of two virulence factors (a toxic protease, AsaP1, and a cytotoxic factor) and a brown pigment were reduced in the mutant. AsaP1 productionwas...... virulence in fish and QS has not previously been associated with A. salmonicida infections in fish. Furthermore, AsaP1 production has not previously been shown to be QS regulated. The simplicity of the A. salmonicida subsp. achromogenes LuxIR-type QS system and the observation that synthetic QSI can inhibit...

  6. Effects of Pistacia atlantica (subsp. Mutica oil extracts on antioxidant activities during experimentally induced cutaneous wound healing in rats

    Directory of Open Access Journals (Sweden)

    Ahmad Reza Hamidi

    2015-01-01

    Full Text Available The fruits of Pistacia atlantica (subsp. mutica have been used traditionally for the treatment of peptic ulcer, as a mouth freshener and have recently been introduced as a source of antioxidant vegetable oils. The aim of this study was to investigate the antioxidant activity of the gel forms, from P. atlantica (subsp. mutica oil extraction on enzymatic antioxidants in experimental wound created in rat. A square-shaped skin defect (2×2 cm was created aseptically by surgical excision at the first thoracic vertebrae. Then animals were randomly allocated in four groups (I, untreated controls; II, topically treated base gel; III, topically treated 5% gel; IV, topically treated 10% gel. Blood sampling was accomplished at 3, 7, 10, 14 and 21 days post-injury. Samples were collected for measuring antioxidant enzymes activities (superoxide dismutase, catalase and glutathione peroxidase activity in red cells and lipid peroxidation (plasma malondialdehyde. The data analysis generally evidenced that the activities of the main antioxidant enzymes began to decrease significantly at 7 days after the wound was created in control and base gel groups. This remarkable decline became more evident in the period between 10 to 21 days post injury but increased progressively in P. atlantica (subsp. mutica treatment groups, especially in gel 10% treatment group during wound healing. The results of this study suggest that excision of the wound leads to oxidative stress and topical administration of P. atlantica (subsp. mutica gels causes remarkable changes in antioxidant parameter during wound closure (especially gel 10% via pro-oxidative, and antioxidant activity can improve oxidative stress.

  7. Streptococcus gallolyticus subsp. pasteurianus en líquido cefalorraquídeo de un paciente pediátrico

    Directory of Open Access Journals (Sweden)

    Liz M Vélez Balestro

    Full Text Available Hasta la fecha se han descrito casos de meningitis por Streptococcus gallolyticus subsp. pasteurianus en adultos, y de los pocos casos pediátricos, el mayor número se presentó en neonatos. En este trabajo presentamos un caso de meningitis y bacteriemia por este estreptococo en un paciente de 9 meses, con reiteradas hospitalizaciones por enfermedades respiratorias; este constituye el primer aislamiento documentado del citado microorganismo en Santa Fe.

  8. New insights into virulence mechanisms of rice pathogen Acidovorax avenae subsp. avenae strain RS-1 following exposure to ?-lactam antibiotics

    OpenAIRE

    Li, Bin; Ge, Mengyu; Zhang, Yang; Wang, Li; Ibrahim, Muhammad; Wang, Yanli; Sun, Guochang; Chen, Gongyou

    2016-01-01

    Recent research has shown that pathogen virulence can be altered by exposure to antibiotics, even when the growth rate is unaffected. Investigating this phenomenon provides new insights into understanding the virulence mechanisms of bacterial pathogens. This study investigates the phenotypic and transcriptomic responses of the rice pathogenic bacterium Acidovorax avenae subsp. avenae (Aaa) strain RS-1 to ?-lactam antibiotics especially Ampicillin (Amp). Our results indicate that exposure to A...

  9. Transcriptome analysis of Acidovorax avenae subsp. avenae cultivated in vivo and co-culture with Burkholderia seminalis

    OpenAIRE

    Bin Li; Muhammad Ibrahim; Mengyu Ge; Zhouqi Cui; Guochang Sun; Fei Xu; Michael Kube

    2014-01-01

    Response of bacterial pathogen to environmental bacteria and its host is critical for understanding of microbial adaption and pathogenesis. Here, we used RNA-Seq to comprehensively and quantitatively assess the transcriptional response of Acidovorax avenae subsp. avenae strain RS-1 cultivated in vitro, in vivo and in co-culture with rice rhizobacterium Burkholderia seminalis R456. Results revealed a slight response to other bacteria, but a strong response to host. In particular, a large numbe...

  10. Modular columns to study depth-dependence behavior of mosquito larvae and toxicity of Bacillus thuringiensis subsp. israelensis.

    Science.gov (United States)

    Meir, Michal; Ben-Dov, Eitan; Zaritsky, Arieh

    2011-03-01

    Modular transparent column system was designed to study depth-dependence behavior of mosquito larvae. The system was used in preliminary experiments to evaluate the effect of water depth on the larvicidal activity of Bacillus thuringiensis subsp. israelensis de Barjac against bottom feeder larvae of Aedes aegypti (Linn.) (Diptera: Culicidae), and suggestions for increasing the efficiency of the device are discussed. Copyright © 2010 Elsevier B.V. All rights reserved.

  11. Molecular analysis and MIRU-VNTR typing of Mycobacterium avium subsp. avium, 'hominissuis' and silvaticum strains of veterinary origin.

    Science.gov (United States)

    Rónai, Zsuzsanna; Csivincsik, Ágnes; Dán, Ádám; Gyuranecz, Miklós

    2016-06-01

    Besides Mycobacterium avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), M. avium subsp. silvaticum (MAS), and 'M. avium subsp. hominissuis' (MAH) are equally important members of M. avium complex, with worldwide distribution and zoonotic potential. Genotypic discrimination is a prerequisite to epidemiological studies which can facilitate disease prevention through revealing infection sources and transmission routes. The primary aim of this study was to identify the genetic diversity within 135 MAA, 62 MAS, and 84 MAH strains isolated from wild and domestic mammals, reptiles and birds. Strains were tested for the presence of large sequence polymorphism LSP(A)17 and were submitted to Mycobacterial interspersed repetitive units-variable-number tandem repeat (MIRU-VNTR) analysis at 8 loci, including MIRU1, 2, 3, and 4, VNTR25, 32, and 259, and MATR9. In 12 strains hsp65 sequence code type was also determined. LSP(A)17 was present only in 19.9% of the strains. All LSP(A)17 positive strains belonged to subspecies MAH. The discriminatory power of the MIRU-VNTR loci set used reached 0.9228. Altogether 54 different genotypes were detected. Within MAH, MAA, and MAS strains 33, 16, and 5 different genotypes were observed. The described genotypes were not restricted to geographic regions or host species, but proved to be subspecies specific. Our knowledge about MAS is limited due to isolation and identification difficulties. This is the first study including a large number of MAS field strains. Our results demonstrate the high diversity of MAH and MAA strains and the relative uniformity of MAS strains. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Cytoembryological aspects of reduced seed setting in Ranunculus ficaria L. subsp. bulbifer (Marsden-Jones Lawalrée

    Directory of Open Access Journals (Sweden)

    Helena Wcisło

    2014-01-01

    Full Text Available Highly disturbed micro- and macro-sporogenesis in R. ficaria L. subsp. bulbifer gives rise to cytologically and genetically unbalanced reproductive cells. In plants from different papulatians, pollen degenerates in various percentages. The majority of apparently normal 7-nucleate ES's undergo abortion. Fertilization occurs sporadically only in ca 7% of ovules. Numerous young achenes undergo degeneration resulting from unviable genotypic combinations. Only 1% of seeds contain multicellular embryos, endosperm tissue and are capable of germination.

  13. Investigating Intraspecific Variation of Acidovorax avenae subsp. citrulli Using DNA Fingerprinting and Whole Cell Fatty Acid Analysis.

    Science.gov (United States)

    Walcott, R R; Langston, D B; Sanders, F H; Gitaitis, R D

    2000-02-01

    ABSTRACT To assess the diversity of Acidovorax avenae subsp. citrulli, 121 strains from watermelon, cantaloupe, and pumpkin were compared using pulse field gel electrophoresis of SpeI-digested DNA and gas chromatographic analysis of fatty acid methyl esters. Twenty-nine unique DNA fragments resulted from DNA digestion, and 14 distinct haplotypes were observed. Based on cluster analysis, two subgroups, I and II, were recognized, which accounted for 84.8% (eight haplotypes) and 15.2% (six haplotypes) of the strains, respectively. Results of cellular fatty acid analysis varied quantitatively and qualitatively for the A. avenae subsp. citrulli strains and supported the existence of the two subgroups. Group I includes strains from cantaloupe and pumpkin as well as the ATCC type strain, which was first described in the United States in 1978, whereas group II represents the typical watermelon fruit blotch-causing strains that appeared in the mainland United States in 1989. Knowledge of the two A. avenae subsp. citrulli groups may be useful in screening for watermelon fruit blotch resistance.

  14. Association of Aeromonas salmonicida subsp. achromogenes in the haemorrhagic blister of cultured carp Cyprinus carpio in West Bengal, India

    Directory of Open Access Journals (Sweden)

    Harresh Adikesavalu

    2014-02-01

    Full Text Available Objective: To study the association of Aeromonas salmonicida subsp. achromogenes in the haemorrhagic blister of cultured exotic carp Cyprinus carpio in West Bengal, India, its cellular fatty acid composition and antibiotic sensitivity. Methods: The phenotypic characterization and antibiotic sensitivity of bacterial flora of diseased fish were performed. The whole cellular fatty acid composition of Aeromonas salmonicida (A. salmonicida was determined by MIS Sherlock automatic identification system. Results: Aeromonas hydrophila, A. salmonicida and Pseudomonas alcaligenes were isolated from the haemorrhagic blister. The fatty acid methyl esters pattern confirmed the A. salmonicida strain as A. salmonicida subsp. achromogenes. Unsaturated fatty acid 16:1 w7c/16:1 w6c (39.09%, saturated fatty acid 16:0 (26.84% and the mono-unsaturated fatty acids 18:1 w7c (8.89% and 16:1 iso I/14:0 3OH (8.49% were the most common fatty acids, which accounted for 83.31% of the total fatty acids. A. salmonicida subsp. achromogenes was highly susceptible to broad spectrum antibiotics such as chloramphenicol, ciprofloxacin, co-trimoxazole, gentamycin, nitrofurantoin and oxytetracycline. Conclusions: The finding on the presence of A. salmonicida in carps necessitates molecular level study on establishing the prevalence of this bacterium in Indian aquaculture systems especially on its free-living viable but non-culturable state.

  15. Functional cream cheese supplemented with Bifidobacterium animalis subsp. lactis DSM 10140 and Lactobacillus reuteri DSM 20016 and prebiotics.

    Science.gov (United States)

    Speranza, Barbara; Campaniello, Daniela; Monacis, Noemi; Bevilacqua, Antonio; Sinigaglia, Milena; Corbo, Maria Rosaria

    2018-06-01

    The aim of this study was to develop a functional fresh cream cheese with Bifidobacterium animalis subsp. lactis DSM 10140 or Lactobacillus reuteri DSM 20016 and prebiotics (inulin, FOS and lactulose). The research was divided into two steps: in vitro evaluation of the effects of prebiotic compounds; validation at laboratory level with production of functional cream mini-cheeses. Prebiotics showed a protective effect: B. animalis subsp. lactis DSM 10140 cultivability on Petri dishes was positively influenced by lactulose, whereas fructooligosaccharides (FOS) were the prebiotic compounds able to prolong Lb. reuteri DSM 20016 cultivability. At 30 °C, a prolongation of the death time (more than 300 days) was observed, while the controls showed death time values about 100 days. At 45 °C, death time values increased from 32.2 (control) to 33, 35, and 38 days in the samples added with FOS, inulin and lactulose, respectively. Lactulose and FOS were chosen to be added to cream mini-cheeses inoculated with B. animalis subsp. lactis DSM 10140 and Lb. reuteri DSM 20016, respectively; the proposed functional cream cheese resulted in a product with favourable conditions for the viability of both probiotics which maintained cultivable cells above the recommended level during 28 days of storage at 4 °C with good sensory characteristics. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Reclassification of the larval pathogen for marine bivalves Vibrio tubiashii subsp. europaeus as Vibrio europaeus sp. nov.

    Science.gov (United States)

    Dubert, Javier; Romalde, Jesús L; Spinard, Edward J; Nelson, David R; Gomez-Chiarri, Marta; Barja, Juan L

    2016-11-01

    The Orientalis clade has a relevant significance for bivalve aquaculture since it includes the pathogens Vibrio bivalvicida, Vibrio tubiashii subsp. tubiashii and Vibrio tubiashii subsp. europaeus. However, the previous taxonomic description of the subspecies of V. tubiashii shows some incongruities that should be emended. In the genomic age, the comparison between genome assemblies is the key to clarify the taxonomic position of both subspecies. With this purpose, we have tested the ability of multilocus sequence analysis based on eight housekeeping gene sequences (gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA and topA), different in silico genome-to-genome comparisons, chemotaxonomic features and phenotypic traits to reclassify the subspecies V. tubiashii subsp. europaeus within the Orientalis clade. This polyphasic approach clearly demonstrated that this subspecies is phylogenetically and phenotypically distinct from V. tubiashii and should be elevated to the rank of species as Vibrio europaeus sp. nov. This reclassification allows us to update the Orientalis clade (V. bivalvicida,V. brasiliensis, V. crosai, V. hepatarius, V. orientalis, V. sinaloensis, V. tubiashii and V. europaeus sp. nov.) and reconstruct a better phylogeny of the genus Vibrio. An emended description of V. tubiashii is provided. Finally, the proposed novel species is represented by emergent bivalve pathogens [type strain PP-638T (=CECT 8136T=DSM 27349T), PP2-843 and 07/118 T2] responsible for high mortalities in Spanish and French hatcheries.

  17. Response surface modeling for the inactivation of Bacillus subtilis subsp. niger spores by chlorine dioxide gas in an enclosed space.

    Science.gov (United States)

    Wang, Tao; Qi, Jiancheng; Wu, Jinhui; Hao, Limei; Yi, Ying; Lin, Song; Zhang, Zongxing

    2016-05-01

    Bacillus subtilis subsp. niger spores are a commonly used biological indicator to evaluate the disinfection of an enclosed space. In the present study, chlorine dioxide (ClO2) gas was applied to inactivate B. subtilis subsp. niger spores in an enclosed space. The effects of the ClO2 gas concentration (1-3 mg/l), relative humidity (RH, 30-70%) and exposure time (30-90 min) were investigated using a response surface methodology (RSM). A three-factor Box-Behnken experimental design was used. The obtained data were adequately fitted to a second-order polynomial model with an R2adj of 0.992. The ClO2 gas concentration, RH and exposure time all significantly (Pgas concentration and RH as well as that between the exposure time and RH indicated significant and synergistic effects (Pgas. The inactivation of indoor biological contaminants plays an important role in preventing the transmission of pathogens and ensuring human safety. The predictive model using response surface methodology indicates the influence and interaction of the main factors on the inactivation of Bacillus subtilis subsp. niger spores by ClO2 gas, and can predict a ClO2 gas treatment condition to achieve an effective sterilization of enclosed spaces. The results in this paper will provide a reference for the application of ClO2 gas treatments for indoor disinfection.

  18. Results of Salmonella enterica subsp. enterica serotype identification by Salmonella Check&Trace microarray in international External Quality Assurance Systems.

    Science.gov (United States)

    Madaczak, Grzegorz; Szych, Jolanta; Wasiak, Monika

    2015-01-01

    Traditionally Salmonella enterica subsp. enterica serotypes are identified by slide agglutination with specific antisera for somatic, flagellar and sometimes capsular antigens. An alternative way is genoserotyping using for example a microarray, eg. commercially available test Check&Trace Salmonella. The goal of this study was to evaluate the Check&Trace Salmonella microarray for Salmonella enterica subsp. enterica serotype identification, using Salmonella strains provided by reference laboratories during External Quality Assurance Systems organized for national reference laboratories by ECDC and WHO GFN. 80 Salmonella enterica subsp. enterica have been tested using Check & Trace Salmonella (Check-Points BC, Netherlands). Also classical slide agglutination was performed according to EN ISO 6579:2003/Al:2007 norm, used as reference method. In the group of 80 tested strains, 66% were identified correctly, 4% gave uncertain results and 29% showed "Salmonella, genovar" without a serotype, of which 69% were not included in the CTS list of serotypes. Finally one strain has been recognized incorrectly. Because of IVD certification lack, the CTS test could not be recommended to clinical laboratories. AOAC-RI and OIE certification for test cause, that CTS could be used in most food, environmental and veterinary laboratories with the condition, that all unrecognized strains should be sent to a reference laboratory, to type according to EN ISO 6579:2003/Al:2007 norm, by KWM serotyping or other equal alternative methods.

  19. Streptococcus equi subsp zooepidemicus pleuropneumonia and peritonitis in a dromedary camel (Camelus dromedarius) calf in North America.

    Science.gov (United States)

    Stoughton, William B; Gold, Jenifer

    2015-08-01

    A 12-week-old female dromedary camel (Camelus dromedarius) calf was evaluated because of acute (Streptococcus equi subsp zooepidemicus septicemia as the etiology for the polyserositis (ie, alpaca fever). Treatment with IV broad-spectrum antimicrobials, an NSAID, and pleural drainage was initiated. Clinical signs of pleuropneumonia, peritonitis, and systemic infection improved rapidly 24 hours after initiation of medical treatment. The calf was discharged from the hospital after 11 days, and antimicrobial treatment continued for 2 weeks after discharge. At follow-up approximately 4 weeks after hospital discharge (6 weeks after the initial examination), there were no clinical signs suggestive of relapse or any reported complications. S equi subsp zooepidemicus may cause polyserositis in Old World camelids (eg, dromedary camels) with signs similar to those seen in New World camelids (eg, alpaca and llama). The rapid response to medical treatment for the patient described suggested that S equi subsp zooepidemicus-induced polyserositis (alpaca fever) in dromedary camels may respond favorably to appropriate treatment. Reducing stress, reducing overcrowding, and separate housing of equids and camelids are suggested. Further studies are needed to better assess the epidemiology of alpaca fever in dromedary camels in North America.

  20. The systematic position of Dryopteris blanfordii subsp. nigrosquamosa (Ching) Fraser-Jenkins within the genusDryopterisAdans.

    Science.gov (United States)

    Krinitsina, Anastasiya A; Belenikin, Maxim S; Churikova, Olga A; Kuptsov, Sergey V; Antipin, Maxim I; Logacheva, Maria D; Speranskaya, Anna S

    2017-01-01

    Dryopteris blanfordii (C.Hope) C.Chr. is a member of the Dryopteridaceae, growing in high altitude Picea or Abies forests (2900-3500 m) in China and India. Phylogenetic relationships between D. blanfordii subsp. nigrosquamosa and closely related species of Dryopteris were investigated using a combined analysis of multiple molecular data sets (the protein-coding region of rbcL and matK genes and intergenic spacers psbA-trnH , trnP-petG , rps4-trnS , trnL-trnF and rbcL-accD ). An assumption about the position of D. blanfordii subsp. nigrosquamosa within Dryopteris was made by using the Maximum Likelihood and Bayesian Inference approach and chloroplast marker sequences of Dryopteris species from GenBank. The results demonstrated that Asian taxa D. blanfordii subsp. nigrosquamosa and D. laeta as well as two American species D. arguta and D. marginalis belong to the same clade, all four of them being part of Dryopteris section Dryopteris.

  1. The systematic position of Dryopteris blanfordii subsp. nigrosquamosa (Ching Fraser-Jenkins within the genus Dryopteris Adans.

    Directory of Open Access Journals (Sweden)

    Anastasiya A. Krinitsina

    2017-11-01

    Full Text Available Dryopteris blanfordii (C.Hope C.Chr. is a member of the Dryopteridaceae, growing in high altitude Picea or Abies forests (2900–3500 m in China and India. Phylogenetic relationships between D. blanfordii subsp. nigrosquamosa and closely related species of Dryopteris were investigated using a combined analysis of multiple molecular data sets (the protein-coding region of rbcL and matK genes and intergenic spacers psbA-trnH, trnP-petG, rps4-trnS, trnL-trnF and rbcL-accD. An assumption about the position of D. blanfordii subsp. nigrosquamosa within Dryopteris was made by using the Maximum Likelihood and Bayesian Inference approach and chloroplast marker sequences of Dryopteris species from GenBank. The results demonstrated that Asian taxa D. blanfordii subsp. nigrosquamosa and D. laeta as well as two American species D. arguta and D. marginalis belong to the same clade, all four of them being part of Dryopteris section Dryopteris.

  2. The Host Genotype and Environment Affect Strain Types of Bifidobacterium longum subsp. longum Inhabiting the Intestinal Tracts of Twins.

    Science.gov (United States)

    Zhang, Min; Hang, Xiaomin; Tan, Jing; Yang, Hong

    2015-07-01

    To investigate the influences of host genotype and environment on Bifidobacterium longum subsp. longum inhabiting human intestines at the strain level, six pairs of twins, divided into two groups (children and adults), were recruited. Each group consisted of two monozygotic (MZ) twin pairs and one dizygotic (DZ) twin pair. Child twins had been living together from birth, while adult twins had been living separately for 5 to 10 years. A total of 345 B. longum subsp. longum isolates obtained from 60 fecal samples from these twins were analyzed by multilocus sequence typing (MLST), and 35 sequence types (STs) were finally acquired. Comparison of strains within and between the twin pairs showed that no strains with identical STs were observed between unrelated individuals or within adult DZ twin pairs. Eight STs were found to be monophyletic, existing within MZ twins and child DZ twins. The similarity of strain types within child cotwins was significantly higher than that within adult cotwins, which indicated that environment was one of the important determinants in B. longum subsp. longum strain types inhabiting human intestines. However, although these differences between MZ and DZ twins were observed, it is still difficult to reach an exact conclusion about the impact of host genotype. This is mainly because of the limited number of subjects tested in the present study and the lack of strain types tracing in the same twin pairs from birth until adulthood. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  3. Isolation and Genomic Characterization of the T4-Like Bacteriophage PM2 Infecting Pectobacterium carotovorum subsp. carotovorum.

    Science.gov (United States)

    Lim, Jeong-A; Lee, Dong Hwan; Heu, Sunggi

    2015-03-01

    In order to control Pectobacterium carotovorum subsp. carotovorum, a novel virulent bacteriophage PM2 was isolated. Bacteriophage PM2 can infect 48% of P. carotovorum subsp. carotovorum and 78% of P. carotovorum subsp. brasilliensis but none of atrosepticum, betavasculorum, odoriferum and wasabiae isolates had been infected with PM2. PM2 phage belongs to the family Myoviridae, and contains a large head and contractile tail. It has a 170,286 base pair genome that encodes 291 open reading frames (ORFs) and 12 tRNAs. Most ORFs in bacteriophage PM2 share a high level of homology with T4-like phages including IME08, RB69, and JS98. Phylogenetic analysis based on the amino acid sequence of terminase large subunits confirmed that PM2 is classified as a T4-like phage. It contains no integrase- or no repressor-coding genes related to the lysogenic cycle, and lifestyle prediction using PHACT software suggested that PM2 is a virulent bacteriophage.

  4. Characterization of genes required for the pathogenicity of Pectobacterium carotovorum subsp. carotovorum Pcc21 in Chinese cabbage.

    Science.gov (United States)

    Lee, Dong Hwan; Lim, Jeong-A; Lee, Juneok; Roh, Eunjung; Jung, Kyusuk; Choi, Minseon; Oh, Changsik; Ryu, Sangryeol; Yun, Jongchul; Heu, Sunggi

    2013-07-01

    Pectobacterium carotovorum subsp. carotovorum is a well-known plant pathogen that causes severe soft rot disease in various crops, resulting in considerable economic loss. To identify pathogenicity-related factors, Chinese cabbage was inoculated with 5314 transposon mutants of P. carotovorum subsp. carotovorum Pcc21 derived using Tn5 transposon mutagenesis. A total of 35 reduced-virulence or avirulent mutants were isolated, and 14 loci were identified. The 14 loci could be functionally grouped into nutrient utilization (pyrD, purH, purD, leuA and serB), production of plant cell-wall-degrading enzymes (PCWDEs) (expI, expR and PCC21_023220), motility (flgA, fliA and flhB), biofilm formation (expI, expR and qseC), susceptibility to antibacterial plant chemicals (tolC) and unknown function (ECA2640). Among the 14 genes identified, qseC, tolC and PCC21_023220 are novel pathogenicity factors of P. carotovorum subsp. carotovorum involved in biofilm formation, phytochemical resistance and PCWDE production, respectively.

  5. Isolation and Genomic Characterization of the T4-Like Bacteriophage PM2 Infecting Pectobacterium carotovorum subsp. carotovorum

    Directory of Open Access Journals (Sweden)

    Jeong-A Lim

    2015-03-01

    Full Text Available In order to control Pectobacterium carotovorum subsp. carotovorum, a novel virulent bacteriophage PM2 was isolated. Bacteriophage PM2 can infect 48% of P. carotovorum subsp. carotovorum and 78% of P. carotovorum subsp. brasilliensis but none of atrosepticum, betavasculorum, odoriferum and wasabiae isolates had been infected with PM2. PM2 phage belongs to the family Myoviridae, and contains a large head and contractile tail. It has a 170,286 base pair genome that encodes 291 open reading frames (ORFs and 12 tRNAs. Most ORFs in bacteriophage PM2 share a high level of homology with T4-like phages including IME08, RB69, and JS98. Phylogenetic analysis based on the amino acid sequence of terminase large subunits confirmed that PM2 is classified as a T4-like phage. It contains no integrase- or no repressor-coding genes related to the lysogenic cycle, and lifestyle prediction using PHACT software suggested that PM2 is a virulent bacteriophage.

  6. Improved trehalose production from biodiesel waste using parent and osmotically sensitive mutant of Propionibacterium freudenreichii subsp. shermanii under aerobic conditions.

    Science.gov (United States)

    Ruhal, Rohit; Choudhury, Bijan

    2012-08-01

    Trehalose is an important nutraceutical of wide commercial interest in the food processing industry. Recently, crude glycerol was reported to be suitable for the production of trehalose using a food microbe, Propionibacterium freudenreichii subsp. shermanii, under static flask conditions. Similarly, enhanced trehalose yield was reported in an osmotically sensitive mutant of the same strain under anaerobic conditions. In the present study, an effort was made to achieve higher production of trehalose, propionic acid, and lactic acid using the parent and an osmotically sensitive mutant of P. freudenreichii subsp. shermanii under aeration conditions. Under aeration conditions (200 rpm in shake flasks and 30 % air saturation in a batch reactor), biomass was increased and approximately 98 % of crude glycerol was consumed. In the parent strain, a trehalose titre of 361 mg/l was achieved, whereas in the mutant strain a trehalose titre of 1.3 g/l was produced in shake flask conditions (200 rpm). In the mutant strain, propionic and lactic acid yields of 0.53 and 0.21 g/g of substrate were also achieved with crude glycerol. Similarly, in controlled batch reactor culturing conditions a final trehalose titre of approximately 1.56 g/l was achieved with the mutant strain using crude glycerol as the substrate. Enhanced production of trehalose using P. freudenreichii subsp. shermanii from waste under aeration conditions is reported here. Higher production of trehalose was not due to a higher yield of trehalose but to a higher final biomass concentration.

  7. Can glandular hair density be a breeding marker for Origanum vulgare subsp. hirtum with high essential oil content?

    Science.gov (United States)

    Szabó, Krisztina; Sárosi, Szilvia; Cserháti, Beatrix; Ferenczy, Antal

    2010-09-01

    Origanum vulgare subsp. hirtum (Link) Ietswaart is an essential oil rich plant traditionally used as oregano. Based on the interest of the essential oil producing sector, in 2000 we have started a breeding program of O. vulgare subsp. hirtum. Plant material for our breeding work consists of 6 progeny. Individual evaluation of the plant material was carried out in 2008-2009 with the primary aim of finding mother plants with appropriate morphological features, high essential oil content (> 7%) and with carvacrol as the main essential oil component. Among the survey of morphological characteristics special attention was given to glandular hair density in order to test the usability of it as a morphological marker for screening progeny for high essential oil content. The characteristics of the progeny can be described with high variability ensuring the possibility of a good selection base. Evaluating the morphology, essential oil content and constitution of the individuals, 20 plants were selected on the grounds of their high (7-8.6%) essential oil content, high ratio (70-93%) of carvacrol in the essential oil and typical morphological features of O. vulgare subsp. hirtum. From the results of glandular hair density it can be stated that the correlation between glandular hair density of the upper, middle and lower leaves either on vegetative or generative shoots and essential oil content was never strong enough (correlation coefficient < or = 0.5) to use it exclusively as a morphological marker for individual selection.

  8. Variable-Number Tandem-Repeat Analysis of Respiratory and Household Water Biofilm Isolates of “Mycobacterium avium subsp. hominissuis” with Establishment of a PCR Database

    Science.gov (United States)

    Iakhiaeva, Elena; Howard, Susan T.; Brown Elliott, Barbara A.; McNulty, Steven; Newman, Kristopher L.; Falkinham, Joseph O.; Williams, Myra; Kwait, Rebecca; Lande, Leah; Vasireddy, Ravikiran; Turenne, Christine

    2016-01-01

    “Mycobacterium avium subsp. hominissuis” is an important cause of pulmonary disease. It is acquired from environmental sources, but there is no methodology for large population studies. We evaluated the potential of variable-number tandem-repeat (VNTR) analysis. Clinical and household biofilm M. avium isolates underwent molecular identification. Testing for IS901 was done to separate M. avium subsp. avium from M. avium subsp. hominissuis. VNTR types were defined using VNTR loci, and subtyping was performed using 3′ hsp65 and internal transcribed spacer (ITS) sequencing. Forty-nine VNTR types and eight subtypes of M. avium subsp. hominissuis (IS901 negative) were identified among 416 isolates of M. avium from 121 patients and 80 biofilm sites. Of those types, 67% were found only among patient isolates, 11% only among household water isolates, and 23% among both. Of 13 VNTR types that included ≥4 patients, the majority (61.5%) represented geographic clustering (same city). Most VNTR types with multiple patients belonged to the same 3′ hsp65 sequence code (sequevar). A total of 44 isolates belonging to four M. avium subsp. hominissuis VNTR types (8%), including three with the rare Mav-F ITS sequence and 0/8 subspecies, produced amplicons with IS901 PCR primers. By sequencing, all 44 amplicons were not IS901 but ISMav6, which was recently observed in Japan but had not been previously described among U.S. isolates. VNTR analysis of M. avium subsp. hominissuis isolates is easier and faster than pulsed-field gel electrophoresis. Seven VNTR loci separated 417 isolates into 49 types. No isolates of M. avium subsp. avium were identified. The distributions of the VNTR copy numbers, the allelic diversity, and the low prevalence of ISMav6 differed from the findings for respiratory isolates reported from Japan. PMID:26739155

  9. The genome of Aeromonas salmonicida subsp. salmonicida A449: insights into the evolution of a fish pathogen

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    Murphy Colleen

    2008-09-01

    Full Text Available Abstract Background Aeromonas salmonicida subsp. salmonicida is a Gram-negative bacterium that is the causative agent of furunculosis, a bacterial septicaemia of salmonid fish. While other species of Aeromonas are opportunistic pathogens or are found in commensal or symbiotic relationships with animal hosts, A. salmonicida subsp. salmonicida causes disease in healthy fish. The genome sequence of A. salmonicida was determined to provide a better understanding of the virulence factors used by this pathogen to infect fish. Results The nucleotide sequences of the A. salmonicida subsp. salmonicida A449 chromosome and two large plasmids are characterized. The chromosome is 4,702,402 bp and encodes 4388 genes, while the two large plasmids are 166,749 and 155,098 bp with 178 and 164 genes, respectively. Notable features are a large inversion in the chromosome and, in one of the large plasmids, the presence of a Tn21 composite transposon containing mercury resistance genes and an In2 integron encoding genes for resistance to streptomycin/spectinomycin, quaternary ammonia compounds, sulphonamides and chloramphenicol. A large number of genes encoding potential virulence factors were identified; however, many appear to be pseudogenes since they contain insertion sequences, frameshifts or in-frame stop codons. A total of 170 pseudogenes and 88 insertion sequences (of ten different types are found in the A. salmonicida genome. Comparison with the A. hydrophila ATCC 7966T genome reveals multiple large inversions in the chromosome as well as an approximately 9% difference in gene content indicating instances of single gene or operon loss or gain. A limited number of the pseudogenes found in A. salmonicida A449 were investigated in other Aeromonas strains and species. While nearly all the pseudogenes tested are present in A. salmonicida subsp. salmonicida strains, only about 25% were found in other A. salmonicida subspecies and none were detected in other

  10. Pollen morphology and its taxonomic significance in the genus Bomarea Mirb. (Alstroemeriaceae - II. Subgenus Bomarea

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    Abul Khayer Mohammad Golam Sarwar

    2015-12-01

    Full Text Available Pollen morphology of 52 species (out of c. 79 of the Bomarea subgenus Bomarea was examined using light microscopy and scanning electron microscopy (SEM, or using SEM alone. The studied species of Bomarea were stenopalynous, characterized by large, oblate, monosulcate monads with reticulate exine sculpture in most species. Wide variation was observed in quantitative palynological features. The studied taxa were divided into four major groups based on exine ornamentation observed under SEM: microreticulate, reticulate, coarsely rugulate, and psilate-perforate. The reticulate exine sculpture may be a plesiomorphic character state for the genus Bomarea, and the coarsely rugulate and finely rugulate-perforate or psilate-perforate exine sculptures may have evolved independently more than once. In agglomerative hierarchical clustering (AHC analyses of the genus Bomarea using quantitative pollen data, the studied species were distributed in either two (similarity-based or four (dissimilarity-based major clusters. Neither the recent molecular phylogenetic analyses nor the AHC analyses of Bomarea have recovered clades/clusters that represent traditionally recognized subgeneric taxa for the genus. Therefore, the most reliable infrageneric classification of Bomarea can be achieved by combining morphological, palynological, and molecular data from more extentive sampling of all the species.

  11. Efecto antitumoral del extracto acuoso de Bomarea cornigera (Alstroemeriaceae en sarcomas inducidos en ratones

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    Christian Villanueva

    2011-05-01

    Full Text Available Se investigó el efecto antitumoral del extracto acuoso del bejuco Bomarea cornigera. Ratones de la cepa Swiss albina fueron inoculados con la línea tumoral TG-180 por 15 días; luego del cual se separaron en 5 grupos (n=5 por grupo. Se administro intraperitonealmente ciclofosfamida (control positivo, agua destilada (control negativo y el extracto en concentraciones de 1X, 2X y 4X; se evaluó la morbilidad, mortalidad, el peso y la longitud del sarcoma. Se encontró un efecto inhibidor del extracto de B. cornigera en el desarrollo del tumor sólido en ratones en los cuales se les transplanto el sarcoma TG-180. Las tasas de inhibición fueron 87,44 y 8,52% después de 17 días de tratamiento considerando la dosis 1X (más baja y 2X (intermedia, respectiva- mente. Estos resultados sugieren que la administración de extracto acuoso de B. cornigera vía intraperitoneal puede ser útil como inhibidor del cáncer.

  12. Micromorphological Characterization of the Leaf and Rhizome of Agapanthus praecox subsp. praecox Willd. (Amaryllidaceae

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    Olubunmi Josephine Sharaibi

    2017-01-01

    Full Text Available Agapanthus praecox subsp. praecox Willd. is a highly valued medicinal plant of family Amaryllidaceae. The genus Agapanthus has been difficult to classify into distinct species due to broad similar morphology of its members. Present taxonomic confusion in this genus and numerous medicinal uses of A. praecox necessitate its proper identification. The leaf and rhizome microcharacters were studied using scanning electron, light microscopy, and energy dispersive X-ray spectroscopy. Epidermal cells are polygonal having wavy anticlinal walls with mean adaxial length of 80.04±0.5 μm and mean abaxial length of 85.33±0.6 μm. The leaf is amphistomatic with anomocytic stomata with mean pore length of 22.14±0.2 μm on the adaxial and 15.02±0.3 μm on the abaxial surface. The mean stomata densities on the adaxial and abaxial surfaces were 280.5±0.5 mm2 and 350.6±0.6 mm2. Trichomes and secretory ducts are absent on both surfaces. EDX spectroscopy showed that beryllium, carbon, oxygen, sodium, and silicon were present on both epidermal surfaces and rhizome while nitrogen, aluminum, and chlorine were detected only on the adaxial surface and sulphur was detected only in the rhizome.

  13. Characterization of a Mycobacterium avium subsp. avium Operon Associated with Virulence and Drug Detoxification

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    Mariana Noelia Viale

    2014-01-01

    Full Text Available The lprG-p55 operon of Mycobacterium tuberculosis and Mycobacterium bovis is involved in the transport of toxic compounds. P55 is an efflux pump that provides resistance to several drugs, while LprG is a lipoprotein that modulates the host's immune response against mycobacteria. The knockout mutation of this operon severely reduces the replication of both mycobacterial species during infection in mice and increases susceptibility to toxic compounds. In order to gain insight into the function of LprG in the Mycobacterium avium complex, in this study, we assayed the effect of the deletion of lprG gene in the D4ER strain of Mycobacterium avium subsp. avium. The replacement of lprG gene with a hygromycin cassette caused a polar effect on the expression of p55. Also, a twofold decrease in ethidium bromide susceptibility was observed and the resistance to the antibiotics rifampicin, amikacin, linezolid, and rifabutin was impaired in the mutant strain. In addition, the mutation decreased the virulence of the bacteria in macrophages in vitro and in a mice model in vivo. These findings clearly indicate that functional LprG and P55 are necessary for the correct transport of toxic compounds and for the survival of MAA in vitro and in vivo.

  14. Inhibitory Effect of Camptothecin against Rice Bacterial Brown Stripe Pathogen Acidovorax avenae subsp. avenae RS-2.

    Science.gov (United States)

    Dong, Qiaolin; Luo, Ju; Qiu, Wen; Cai, Li; Anjum, Syed Ishtiaq; Li, Bin; Hou, Mingsheng; Xie, Guanlin; Sun, Guochang

    2016-07-27

    Camptothecin (CPT) has anticancer, antiviral, and antifungal properties. However, there is a dearth of information about antibacterial activity of CPT. Therefore, in this study, we investigated the inhibitory effect of CPT on Acidovorax avenae subsp. avenae strain RS-2, the pathogen of rice bacterial brown stripe, by measuring cell growth, DNA damage, cell membrane integrity, the expression of secretion systems, and topoisomerase-related genes, as well as the secretion of effector protein Hcp. Results indicated that CPT solutions at 0.05, 0.25, and 0.50 mg/mL inhibited the growth of strain RS-2 in vitro, while the inhibitory efficiency increased with an increase in CPT concentration, pH, and incubation time. Furthermore, CPT treatment affected bacterial growth and replication by causing membrane damage, which was evidenced by transmission electron microscopic observation and live/dead cell staining. In addition, quantitative real-time PCR analysis indicated that CPT treatment caused differential expression of eight secretion system-related genes and one topoisomerase-related gene, while the up-regulated expression of hcp could be justified by the increased secretion of Hcp based on the ELISA test. Overall, this study indicated that CPT has the potential to control the bacterial brown stripe pathogen of rice.

  15. Sugarcane transcriptome analysis in response to infection caused by Acidovorax avenae subsp. avenae.

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    Ailton B Santa Brigida

    Full Text Available Sugarcane is an important tropical crop mainly cultivated to produce ethanol and sugar. Crop productivity is negatively affected by Acidovorax avenae subsp avenae (Aaa, which causes the red stripe disease. Little is known about the molecular mechanisms triggered in response to the infection. We have investigated the molecular mechanism activated in sugarcane using a RNA-seq approach. We have produced a de novo transcriptome assembly (TR7 from sugarcane RNA-seq libraries submitted to drought and infection with Aaa. Together, these libraries present 247 million of raw reads and resulted in 168,767 reference transcripts. Mapping in TR7 of reads obtained from infected libraries, revealed 798 differentially expressed transcripts, of which 723 were annotated, corresponding to 467 genes. GO and KEGG enrichment analysis showed that several metabolic pathways, such as code for proteins response to stress, metabolism of carbohydrates, processes of transcription and translation of proteins, amino acid metabolism and biosynthesis of secondary metabolites were significantly regulated in sugarcane. Differential analysis revealed that genes in the biosynthetic pathways of ET and JA PRRs, oxidative burst genes, NBS-LRR genes, cell wall fortification genes, SAR induced genes and pathogenesis-related genes (PR were upregulated. In addition, 20 genes were validated by RT-qPCR. Together, these data contribute to a better understanding of the molecular mechanisms triggered by the Aaa in sugarcane and opens the opportunity for the development of molecular markers associated with disease tolerance in breeding programs.

  16. Inhibitory Effect of Camptothecin against Rice Bacterial Brown Stripe Pathogen Acidovorax avenae subsp. avenae RS-2

    Directory of Open Access Journals (Sweden)

    Qiaolin Dong

    2016-07-01

    Full Text Available Camptothecin (CPT has anticancer, antiviral, and antifungal properties. However, there is a dearth of information about antibacterial activity of CPT. Therefore, in this study, we investigated the inhibitory effect of CPT on Acidovorax avenae subsp. avenae strain RS-2, the pathogen of rice bacterial brown stripe, by measuring cell growth, DNA damage, cell membrane integrity, the expression of secretion systems, and topoisomerase-related genes, as well as the secretion of effector protein Hcp. Results indicated that CPT solutions at 0.05, 0.25, and 0.50 mg/mL inhibited the growth of strain RS-2 in vitro, while the inhibitory efficiency increased with an increase in CPT concentration, pH, and incubation time. Furthermore, CPT treatment affected bacterial growth and replication by causing membrane damage, which was evidenced by transmission electron microscopic observation and live/dead cell staining. In addition, quantitative real-time PCR analysis indicated that CPT treatment caused differential expression of eight secretion system-related genes and one topoisomerase-related gene, while the up-regulated expression of hcp could be justified by the increased secretion of Hcp based on the ELISA test. Overall, this study indicated that CPT has the potential to control the bacterial brown stripe pathogen of rice.

  17. Sugarcane transcriptome analysis in response to infection caused by Acidovorax avenae subsp. avenae.

    Science.gov (United States)

    Santa Brigida, Ailton B; Rojas, Cristian A; Grativol, Clícia; de Armas, Elvismary M; Entenza, Júlio O P; Thiebaut, Flávia; Lima, Marcelo de F; Farrinelli, Laurent; Hemerly, Adriana S; Lifschitz, Sérgio; Ferreira, Paulo C G

    2016-01-01

    Sugarcane is an important tropical crop mainly cultivated to produce ethanol and sugar. Crop productivity is negatively affected by Acidovorax avenae subsp avenae (Aaa), which causes the red stripe disease. Little is known about the molecular mechanisms triggered in response to the infection. We have investigated the molecular mechanism activated in sugarcane using a RNA-seq approach. We have produced a de novo transcriptome assembly (TR7) from sugarcane RNA-seq libraries submitted to drought and infection with Aaa. Together, these libraries present 247 million of raw reads and resulted in 168,767 reference transcripts. Mapping in TR7 of reads obtained from infected libraries, revealed 798 differentially expressed transcripts, of which 723 were annotated, corresponding to 467 genes. GO and KEGG enrichment analysis showed that several metabolic pathways, such as code for proteins response to stress, metabolism of carbohydrates, processes of transcription and translation of proteins, amino acid metabolism and biosynthesis of secondary metabolites were significantly regulated in sugarcane. Differential analysis revealed that genes in the biosynthetic pathways of ET and JA PRRs, oxidative burst genes, NBS-LRR genes, cell wall fortification genes, SAR induced genes and pathogenesis-related genes (PR) were upregulated. In addition, 20 genes were validated by RT-qPCR. Together, these data contribute to a better understanding of the molecular mechanisms triggered by the Aaa in sugarcane and opens the opportunity for the development of molecular markers associated with disease tolerance in breeding programs.

  18. Studies on the biology of the crisamicin-producing organism Micromonospora purpureochromogenes subsp. celinoensis

    International Nuclear Information System (INIS)

    Pope, J.A. Jr.

    1985-01-01

    A Micromonospora isolate, RV-101, obtained from the Red V Coconut Company in the Philippines, produces a new complex of antibiotics, the crisamicins. Using standard taxonomic methods for the genus Micromonospora, including micromorphology, growth characteristics on select media, whole cell analysis of chemical composition, and carbohydrate utilization patterns, the organism was classified as Micromonospora purpureochromogenes subsp. celinoensis. The chief character used in this classification was the production of a dark-brown diffusible pigment on media containing complex sources of nitrogen. The biosynthesis of crisamicin A was investigated by the technique of 13 C acetate feeding and 13 C NMR spectroscopy. Crisamicin A was found to be synthesized from acetate via the polyketide biosynthetic pathway. In addition, the assignment of one of two possible structures, differing in the position of phenolic hydroxyl groups, and in the point of linkage between the two monomers of the molecule, was made using the labeling data. The structure determined demonstrates that crisamicin A is unique among the benzoisochromanequinone antibiotics, in that it lacks an oxygen atom at position C-8

  19. Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil.

    Science.gov (United States)

    Albuquerque, Pedro Paulo Feitosa de; Santos, André de Souza; Souza Neto, Orestes Luiz de; Kim, Pomy de Cássia Peixoto; Cavalcanti, Erika Fernanda Torres Samico Fernandes; Oliveira, Júnior Mário Baltazar de; Mota, Rinaldo Aparecido; Júnior, José Wilton Pinheiro

    The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples. Copyright © 2016. Published by Elsevier Editora Ltda.

  20. Development of healthy whey drink with Lactobacillus rhamnosus, Bifidobacterium bifidum and Propionibacterium freudenreichii subsp. shermanii

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    T.K. Maity

    2008-12-01

    Full Text Available Whey beverage was prepared by utilizing Lactobacillus rhamnosus NCDO 243, Bifidobacterium bifidum NCDO 2715 and Propionibacterium freudenreichii subsp. shermanii MTCC 1371 in order to make a fermented probiotic healthy drink. The product made with 4 % mixed culture (1:1:1 inoculated (initial count - lactobacilli 6.2 x 107 CFU/mL, bifidobacteria 5.4 x 107 CFU/mL, propionibacteria 3.9 x 107 CFU/mL in deprotienized whey (4.6 % lactose, 0.62 % ash, 0.48 % fat and 0.5 % protein adjusted to pH 6.4 and incubated at 37 °C for 8 h has a good technological and dietetic criteria required for a probiotic product. Total bacterial count, lactobacilli count, bifidobacteria count, propionibacteria count, titratable acidity, β-D galactosidase activity, concentration of lactic acid and sensory properties were monitored during storage period. The whey beverage fermented for 8 h and prepared with 4 % inoculum of mixed culture (1:1:1 met the probiotic criterion by maintaining each type of bacterial population at counts greater than 108 CFU/mL up to 10 days of storage period. The titratable acidity as well as sensory properties did not change appreciably during first 7 days of storage. At the end of 15 days of storage, slight acidification was detected, although the beverage still retained an acceptable flavour.

  1. Pilot Plant Production of Lactic acid by Lactobacillus casei subsp. casei

    Directory of Open Access Journals (Sweden)

    S.S Mirdamadi

    2012-02-01

    Full Text Available The aim of present study, was to scale up the production of L (+ lactic acid from the laboratory to pilot plant using Lactobacillus casei subsp. casei PTCC 1608. Moreover, the minimum inhibitory concentration of the produced lactic acid and sodium lactate against 4 test strains including Staphylococcus aureus PTCC 1113, Microccoccus luteus PTTC 1110, Escherichia coli PTCC 1330 and Listeria monocytogenes PTCC 1304 were evaluated. According to the results, the specific growth rate of each test strain was decreased by lactic acid. The inhibitory effect of the sodium lactate was lower than lactic acid in all of the experiments. The best carbon (glucose, lactose and whey and nitrogen (corn steep powder sources were optimized in batch and fed batch system and also pH, temperature and aeration were improved in shake flask incubator, 20 l and 750 l stirred tank reactors (STR. Glucose (80 g/l supplemented with (50 g/l whey was found as the best production medium.Productivity and yield of calcium lactate production in laboratory scale were 0.51 g/lh and 0.56%, respectively. Fed batch production of calcium lactate in 20 l bioreactor increased the productivity and yield up to 2.47 and 0.83%. Production and productivity was increased up to 350 g/l and 5.4 g/lh, respectively in scaled up processes by 750 liters bioreactor (STR.

  2. Leaching of Mycobacterium avium subsp. paratuberculosis in Soil under In Vitro Conditions

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    Eran A. Raizman

    2011-01-01

    Full Text Available Mycobacterium avium subsp paratuberculosis (Map, the causative agent of Johne's disease, has a robust ability to survive in the environment. However, the ability of Map to migrate through soil to drainage tiles or ground water, leave the farm, and leak into local watersheds is inadequately documented. In order to assess the ability of Map to leach through soil, two laboratory experiments were conducted. In the first study, 8 columns (30 cm long each of a sandy loam soil were treated with pure cultures of Map. Two soil moisture levels and two Map concentrations were used. The columns were leached with 500 mL of water once a week for three weeks, the leachate was collected, and detection analysis was conducted. In the second experiment, manure from Map negative cows (control and Map high shedder cows (treatment were deposited on 8 similar columns and the columns were leached with 500 mL of water once a week for four weeks. Map detection and numeration in leachate samples were done with RT-PCR and culture techniques, respectively. Using RT-PCR, Map could be detected in the leachates in both experiments for several weeks but could only be recovered using culture techniques in experiment one. Combined, these experiments indicate the potential for Map to move through soil as a result of rainfall or irrigation following application.

  3. The Science behind the Probiotic Strain Bifidobacterium animalis subsp. lactis BB-12(®).

    Science.gov (United States)

    Jungersen, Mikkel; Wind, Anette; Johansen, Eric; Christensen, Jeffrey E; Stuer-Lauridsen, Birgitte; Eskesen, Dorte

    2014-03-28

    This review presents selected data on the probiotic strain Bifidobacterium animalis subsp. lactis BB-12(®) (BB-12(®)), which is the world's most documented probiotic Bifidobacterium. It is described in more than 300 scientific publications out of which more than 130 are publications of human clinical studies. The complete genome sequence of BB-12(®) has been determined and published. BB-12(®) originates from Chr. Hansen's collection of dairy cultures and has high stability in foods and as freeze dried powders. Strain characteristics and mechanisms of BB-12(®) have been established through extensive in vitro testing. BB-12(®) exhibits excellent gastric acid and bile tolerance; it contains bile salt hydrolase, and has strong mucus adherence properties, all valuable probiotic characteristics. Pathogen inhibition, barrier function enhancement, and immune interactions are mechanisms that all have been demonstrated for BB-12(®). BB-12(®) has proven its beneficial health effect in numerous clinical studies within gastrointestinal health and immune function. Clinical studies have demonstrated survival of BB-12(®) through the gastrointestinal tract and BB-12(®) has been shown to support a healthy gastrointestinal microbiota. Furthermore, BB-12(®) has been shown to improve bowel function, to have a protective effect against diarrhea, and to reduce side effects of antibiotic treatment, such as antibiotic-associated diarrhea. In terms of immune function, clinical studies have shown that BB-12(®) increases the body's resistance to common respiratory infections as well as reduces the incidence of acute respiratory tract infections.

  4. Leaching of Mycobacterium avium Subsp paratuberculosis in Soil under In Vitro Conditions.

    Science.gov (United States)

    Raizman, Eran A; Habteselassie, Mussie Y; Wu, Ching C; Lin, Tsang L; Negron, M; Turco, Ronald F

    2011-01-01

    Mycobacterium avium subsp paratuberculosis (Map), the causative agent of Johne's disease, has a robust ability to survive in the environment. However, the ability of Map to migrate through soil to drainage tiles or ground water, leave the farm, and leak into local watersheds is inadequately documented. In order to assess the ability of Map to leach through soil, two laboratory experiments were conducted. In the first study, 8 columns (30 cm long each) of a sandy loam soil were treated with pure cultures of Map. Two soil moisture levels and two Map concentrations were used. The columns were leached with 500 mL of water once a week for three weeks, the leachate was collected, and detection analysis was conducted. In the second experiment, manure from Map negative cows (control) and Map high shedder cows (treatment) were deposited on 8 similar columns and the columns were leached with 500 mL of water once a week for four weeks. Map detection and numeration in leachate samples were done with RT-PCR and culture techniques, respectively. Using RT-PCR, Map could be detected in the leachates in both experiments for several weeks but could only be recovered using culture techniques in experiment one. Combined, these experiments indicate the potential for Map to move through soil as a result of rainfall or irrigation following application.

  5. Cytotoxicity Analysis of Three Bacillus thuringiensis Subsp. israelensis δ-Endotoxins towards Insect and Mammalian Cells

    Science.gov (United States)

    Teixeira Corrêa, Roberto Franco; Ardisson-Araújo, Daniel Mendes Pereira; Monnerat, Rose Gomes; Ribeiro, Bergmann Morais

    2012-01-01

    Three members of the δ-endotoxin group of toxins expressed by Bacillus thuringiensis subsp. israelensis, Cyt2Ba, Cry4Aa and Cry11A, were individually expressed in recombinant acrystalliferous B. thuringiensis strains for in vitro evaluation of their toxic activities against insect and mammalian cell lines. Both Cry4Aa and Cry11A toxins, activated with either trypsin or Spodoptera frugiperda gastric juice (GJ), resulted in different cleavage patterns for the activated toxins as seen by SDS-PAGE. The GJ-processed proteins were not cytotoxic to insect cell cultures. On the other hand, the combination of the trypsin-activated Cry4Aa and Cry11A toxins yielded the highest levels of cytotoxicity to all insect cells tested. The combination of activated Cyt2Ba and Cry11A also showed higher toxic activity than that of toxins activated individually. When activated Cry4Aa, Cry11A and Cyt2Ba were used simultaneously in the same assay a decrease in toxic activity was observed in all insect cells tested. No toxic effect was observed for the trypsin-activated Cry toxins in mammalian cells, but activated Cyt2Ba was toxic to human breast cancer cells (MCF-7) when tested at 20 µg/mL. PMID:23029407

  6. Safety of Bifidobacterium animalis Subsp. Lactis (B. lactis) Strain BB-12-Supplemented Yogurt in Healthy Children.

    Science.gov (United States)

    Tan, Tina P; Ba, Zhaoyong; Sanders, Mary E; D'Amico, Frank J; Roberts, Robert F; Smith, Keisha H; Merenstein, Daniel J

    2017-02-01

    Probiotics are live microorganisms that may provide health benefits to the individual when consumed in sufficient quantities. For studies conducted on health or disease endpoints on probiotics in the United States, the Food and Administration has required those studies to be conducted as investigational new drugs. This phase I, double-blinded, randomized, controlled safety study represents the first requirement of this pathway. The purpose of the study was to determine the safety of Bifidobacterium animalis subsp. lactis (B lactis) strain BB-12 (BB-12)-supplemented yogurt when consumed by a generally healthy group of children. The secondary aim was to assess the effect of BB-12-supplemented yogurt on the gut microbiota of the children. Sixty children ages 1 to 5 years were randomly assigned to consume 4 ounces of either BB-12-supplemented yogurt or nonsupplemented control yogurt daily for 10 days. The primary outcome was to assess safety and tolerability, as determined by the number of reported adverse events. A total of 186 nonserious adverse events were reported, with no significant differences between the control and BB-12 groups. No significant changes due to probiotic treatment were observed in the gut microbiota of the study cohort. BB-12-supplemented yogurt is safe and well-tolerated when consumed by healthy children. The present study will form the basis for future randomized clinical trials investigating the potential effects of BB-12-supplemented yogurt in different disease states.

  7. Chemical composition and antioxidant activity of Origanum vulgare subsp. vulgare essential oil from Iran

    Directory of Open Access Journals (Sweden)

    M. Vazirian

    2015-12-01

    Full Text Available Background and Objectives: Essential oils are very complex mixture of components and their composition may vary in different species or varieties or even within the same variety. Origanum vulgare L. subsp. vulgare is one of the most distributed subspecies within the genus Origanum and has been found to be a poor-oil, categorized in cymyl, bornane or sabinyl chemotypes with higher proportion of sesquiterpenes. In this experiment, the Iranian sample was studied for the chemical composition of the oil and evaluation of its antioxidant activity. Methods: Essential oil was obtained by hydro-distillation and analyzed by GC/MS for determination of components. Antioxidant activity was evaluated by radical scavenging ability (DPPH method and reducing power (FRAP assay. Results: The sample belonged to “thymol” chemotype with the main components as thymol (37.13%, gama-terpinene (9.67%, carvacrol (9.57%, carvacrol methyl ether (6.88, cis-alpha-bisabolene (6.80%, eucalyptol (3.82%, p-cymene (3.58% and elemol (2.04%. The oil of plant showed very strong antioxidant activity (IC50=2.5 µg/mL in DPPH method, which was stronger than the standard antioxidants (Vit E and BHA, p

  8. Structural investigation of cell wall polysaccharides of Lactobacillus delbrueckii subsp. bulgaricus 17.

    Science.gov (United States)

    Vinogradov, E; Sadovskaya, I; Cornelissen, A; van Sinderen, D

    2015-09-02

    Lactobacilli are valuable strains for commercial (functional) food fermentations. Their cell surface-associated polysaccharides (sPSs) possess important functional properties, such as acting as receptors for bacteriophages (bacterial viruses), influencing autolytic characteristics and providing protection against antimicrobial peptides. The current report provides an elaborate molecular description of several surface carbohydrates of Lactobacillus delbrueckii subsp. bulgaricus strain 17. The cell surface of this strain was shown to contain short chain poly(glycerophosphate) teichoic acids and at least two different sPSs, designated here as sPS1 and sPS2, whose chemical structures were examined by 2D nuclear magnetic resonance spectroscopy and methylation analysis. Neutral branched sPS1, extracted with n-butanol, was shown to be composed of hexasaccharide repeating units (-[α-d-Glcp-(1-3)-]-4-β-l-Rhap2OAc-4-β-d-Glcp-[α-d-Galp-(1-3)]-4-α-Rhap-3-α-d-Galp-), while the major component of the TCA-extracted sPS2 was demonstrated to be a linear d-galactan with the repeating unit structure being (-[Gro-3P-(1-6)-]-3-β-Galf-3-α-Galp-2-β-Galf-6-β-Galf-3-β-Galp-). Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Streptococcus thermophilus urease activity boosts Lactobacillus delbrueckii subsp. bulgaricus homolactic fermentation.

    Science.gov (United States)

    Arioli, Stefania; Della Scala, Giulia; Remagni, Maria Chiara; Stuknyte, Milda; Colombo, Stefano; Guglielmetti, Simone; De Noni, Ivano; Ragg, Enzio; Mora, Diego

    2017-04-17

    The proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in the yogurt consortium enhances the growth rate and size of each population. In contrast, the independent growth of the two species in milk leads to a slower growth rate and a smaller population size. In this study, we report the first evidence that the urease activity of S. thermophilus increases the intracellular pH of L. delbrueckii in the absence of carbon source. However, in milk, in the presence of lactose the alkalizing effect of urea-derived ammonia was not detectable. Nevertheless, based on glucose consumption and lactic acid production at different pH in , L. delbrueckii showed an optimum of glycolysis and homolactic fermentation at alkaline pH values. In milk, we observed that ammonia provided by urea hydrolysis boosted lactic acid production in S. thermophilus and in L. delbrueckii when the species were grown alone or in combination. Therefore, we propose that urease activity acts as an altruistic cooperative trait, which is costly for urease-positive individuals but provides a local benefit because other individuals can take advantage of urease-dependent ammonia release. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Subcellular membrane fluidity of Lactobacillus delbrueckii subsp. bulgaricus under cold and osmotic stress.

    Science.gov (United States)

    Meneghel, Julie; Passot, Stéphanie; Cenard, Stéphanie; Réfrégiers, Matthieu; Jamme, Frédéric; Fonseca, Fernanda

    2017-09-01

    Cryopreservation of lactic acid bacteria may lead to undesirable cell death and functionality losses. The membrane is the first target for cell injury and plays a key role in bacterial cryotolerance. This work aimed at investigating at a subcellular resolution the membrane fluidity of two populations of Lactobacillus delbrueckii subsp. bulgaricus when subjected to cold and osmotic stresses associated to freezing. Cells were cultivated at 42 °C in mild whey medium, and they were exposed to sucrose solutions of different osmolarities (300 and 1800 mOsm L -1 ) after harvest. Synchrotron fluorescence microscopy was used to measure membrane fluidity of cells labeled with the cytoplasmic membrane probe 1-[4 (trimethylamino) phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). Images were acquired at 25 and 0 °C, and more than a thousand cells were individually analyzed. Results revealed that a bacterial population characterized by high membrane fluidity and a homogeneous distribution of fluidity values appeared to be positively related to freeze-thaw resistance. Furthermore, rigid domains with different anisotropy values were observed and the occurrence of these domains was more important in the freeze-sensitive bacterial population. The freeze-sensitive cells exhibited a broadening of existing highly rigid lipid domains with osmotic stress. The enlargement of domains might be ascribed to the interaction of sucrose with membrane phospholipids, leading to membrane disorganization and cell degradation.

  11. Galacto-oligosaccharides as protective molecules in the preservation of Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Tymczyszyn, E Elizabeth; Gerbino, Esteban; Illanes, Andrés; Gómez-Zavaglia, Andrea

    2011-04-01

    In this work, the protective capacity of galacto-oligosaccharides in the preservation of Lactobacillus delbrueckii subsp. bulgaricus CIDCA 333 was evaluated. Lactobacillus bulgaricus was freeze-dried or dried over silica gel in the presence of three commercial products containing galacto-oligosaccharides. The freeze-dried samples were stored at 5 and 25°C for different periods of time. After desiccation, freeze-drying or storage, samples were rehydrated and bacterial plate counts were determined. According to the results obtained, all galacto-oligosaccharides assays demonstrated to be highly efficient in the preservation of L. bulgaricus. The higher content of galacto-oligosaccharides in the commercial products was correlated with their higher protective capacity. Galacto-oligosaccharides are widely known by their prebiotic properties. However, their role as protective molecules have not been reported nor properly explored up to now. In this work the protective capacity of galacto-oligosaccharides in the preservation of L. bulgaricus, a strain particularly sensitive to any preservation process, was demonstrated. The novel role of galacto-oligosaccharides as protective molecules opens up several perspectives in regard to their applications. The supplementation of probiotics with galacto-oligosaccharides allows the production of self-protected synbiotic products, galacto-oligosaccharides exerting both a prebiotic and protecting effect. Copyright © 2011 Elsevier Inc. All rights reserved.

  12. Identification of Quorum Sensing Signal Molecule of Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Pang, Xiaoyang; Liu, Cuiping; Lyu, Pengcheng; Zhang, Shuwen; Liu, Lu; Lu, Jing; Ma, Changlu; Lv, Jiaping

    2016-12-14

    Many bacteria in nature use quorum sensing (QS) to regulate gene expression. The quorum sensing system plays critical roles in the adaptation of bacteria to the surrounding environment. Previous studies have shown that during high-density fermentation, the autolysis of lactic acid bacteria was regulated by the QS system, and the two-component system (TCS, LBUL_RS00115/LBUL_RS00110) is involved in the autolysis of Lactobacillus delbrueckii subsp. bulgaricus. However, the QS signal molecule, which regulates this pathway, has not been identified. In this study, we compared the genome of Lactobacillus bulgaricus ATCC BAA-365 with the locus of seven lactobacillus QS systems; the position of the QS signal molecule of Lactobacillus bulgaricus ATCC BAA-365 was predicted by bioinformatics tool. Its function was identified by in vitro experiments. Construction of TCS mutant by gene knockout of LBUL_RS00115 confirmed that the signal molecule regulates the density of the flora by the TCS (LBUL_RS00115/LBUL_RS00110). This study indicated that quorum quenching and inhibition based on the signal molecule might serve as an approach to reduce the rate of autolysis of LAB and increase the number of live bacteria in fermentation.

  13. A two component system is involved in acid adaptation of Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Cui, Yanhua; Liu, Wei; Qu, Xiaojun; Chen, Zhangting; Zhang, Xu; Liu, Tong; Zhang, Lanwei

    2012-05-20

    The Gram-positive bacterium Lactobacillus delbrueckii subsp. bulgaricus is of vital importance to the food industry, especially to the dairy industry. Two component systems (TCSs) are one of the most important mechanisms for environmental sensing and signal transduction in the majority of Gram-positive and Gram-negative bacteria. A typical TCS consists of a histidine protein kinase (HPK) and a cytoplasmic response regulator (RR). To investigate the functions of TCSs during acid adaptation in L. bulgaricus, we used quantitative PCR to reveal how TCSs expression changes during acid adaptation. Two TCSs (JN675228/JN675229 and JN675230/JN675231) and two HPKs (JN675236 and JN675240) were induced during acid adaptation. These TCSs were speculated to be related with the acid adaptation ability of L. bulgaricus. The mutants of JN675228/JN675229 were constructed in order to investigate the functions of JN675228/JN675229. The mutants showed reduced acid adaptation compared to that of wild type, and the complemented strains were similar to the wild-type strain. These observations suggested that JN675228 and JN675229 were involved in acid adaptation in L. bulgaricus. The interaction between JN675228 and JN675229 was identified by means of yeast two-hybrid system. The results indicated there is interaction between JN675228 and JN675229. Crown Copyright © 2011. Published by Elsevier GmbH. All rights reserved.

  14. Physiological Study of Lactobacillus delbrueckii subsp. bulgaricus Strains in a Novel Chemically Defined Medium

    Science.gov (United States)

    Chervaux, Christian; Ehrlich, S. Dusko; Maguin, Emmanuelle

    2000-01-01

    We developed a chemically defined medium called milieu proche du lait (MPL), in which 22 Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) strains exhibited growth rates ranging from 0.55 to 1 h−1. MPL can also be used for cultivation of other lactobacilli and Streptococcus thermophilus. The growth characteristics of L. bulgaricus in MPL containing different carbon sources were determined, including an initial characterization of the phosphotransferase system transporters involved. For the 22 tested strains, growth on lactose was faster than on glucose, mannose, and fructose. Lactose concentrations below 0.4% were limiting for growth. We isolated 2-deoxyglucose-resistant mutants from strains CNRZ397 and ATCC 11842. CNRZ397-derived mutants were all deficient for glucose, fructose, and mannose utilization, indicating that these three sugars are probably transported via a unique mannose-specific-enzyme-II-like transporter. In contrast, mutants of ATCC 11842 exhibited diverse phenotypes, suggesting that multiple transporters may exist in that strain. We also developed a protein labeling method and verified that exopolysaccharide production and phage infection can occur in MPL. The MPL medium should thus be useful in conducting physiological studies of L. bulgaricus and other lactic acid bacteria under well controlled nutritional conditions. PMID:11097906

  15. Detection and Characterization of Histamine-Producing Strains of Photobacterium damselae subsp. damselae Isolated from Mullets.

    Science.gov (United States)

    Trevisani, Marcello; Mancusi, Rocco; Cecchini, Matilde; Costanza, Claudia; Prearo, Marino

    2017-06-20

    Photobacterium damselae subsp. damselae ( Pdd ) is considered to be an emerging pathogen of marine fish and has also been implicated in cases of histamine food poisoning. In this study, eight strains isolated from mullets of the genera Mugil and Liza captured in the Ligurian Sea were characterized, and a method to detect histamine-producing Pdd from fish samples was developed. The histamine-producing potential of the strains was evaluated in culture media (TSB+) using a histamine biosensor. Subsequently, two strains were used to contaminate mackerel fillets (4 or 40 CFU/g), simulating a cross-contamination on the selling fish stalls. Sample homogenates were enriched in TSB+. The cultures were then inoculated on thiosulfate-citrate-bile salts-sucrose agar (TCBS) and the dark green colonies were cultured on Niven agar. The violet isolates were characterized using specific biochemical and PCR based tests. All Pdd strains were histamine producers, yielding concentration varying from 167 and 8977 µg/mL in TSB+ cultures incubated at 30 °C for 24 h. Pdd colonies were detected from the inoculated mackerel samples and their histidine decarboxylase gene was amplified using species-specific primer pairs designed for this study. The results indicate that mullets can be source of Pdd and the fish retailers needs to evaluate the risk posed by cross-contamination on the selling fish stalls.

  16. Mycobacterium avium subsp. paratuberculosis in an Italian Cohort of Type 1 Diabetes Pediatric Patients

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    Maria Luisa Manca Bitti

    2012-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP is the etiological agent of Johne’s disease in ruminants. Recent studies have linked MAP to type 1 diabetes (T1D in the Sardinian population. The aim of this study was to investigate the prevalence of MAP infection in a T1D cohort from continental Italy compared with healthy control subjects. 247 T1D subjects and 110 healthy controls were tested for the presence of MAP. MAP DNA was detected using IS900-specific polymerase chain reaction (PCR. The presence of antibodies towards a MAP antigen, heparin binding hemoagglutinin (HBHA, was detected by ELISA. We demonstrated a higher MAP DNA prevalence in plasma samples from T1D patients and a stronger immune response towards MAP HBHA, compared with healthy control subjects. Moreover, in the recent onset patients, we observed an association between anti-MAP antibodies and HLA DQ2 (DQA1 0201/DQB1 0202. These findings taken together support the hypothesis of MAP as an environmental risk factor for the development of T1D in genetically predisposed subjects, probably involving a mechanism of molecular mimicry between MAP antigens and pancreatic islet β-cells.

  17. Quantitative Proteomics for the Comprehensive Analysis of Stress Responses of Lactobacillus paracasei subsp. paracasei F19.

    Science.gov (United States)

    Schott, Ann-Sophie; Behr, Jürgen; Geißler, Andreas J; Kuster, Bernhard; Hahne, Hannes; Vogel, Rudi F

    2017-10-06

    Lactic acid bacteria are broadly employed as starter cultures in the manufacture of foods. Upon technological preparation, they are confronted with drying stress that amalgamates numerous stress conditions resulting in losses of fitness and survival. To better understand and differentiate physiological stress responses, discover general and specific markers for the investigated stress conditions, and predict optimal preconditioning for starter cultures, we performed a comprehensive genomic and quantitative proteomic analysis of a commonly used model system, Lactobacillus paracasei subsp. paracasei TMW 1.1434 (isogenic with F19) under 11 typical stress conditions, including among others oxidative, osmotic, pH, and pressure stress. We identified and quantified >1900 proteins in triplicate analyses, representing 65% of all genes encoded in the genome. The identified genes were thoroughly annotated in terms of subcellular localization prediction and biological functions, suggesting unbiased and comprehensive proteome coverage. In total, 427 proteins were significantly differentially expressed in at least one condition. Most notably, our analysis suggests that optimal preconditioning toward drying was predicted to be alkaline and high-pressure stress preconditioning. Taken together, we believe the presented strategy may serve as a prototypic example for the analysis and utility of employing quantitative-mass-spectrometry-based proteomics to study bacterial physiology.

  18. Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

    Directory of Open Access Journals (Sweden)

    Christopher D Johnston

    2014-09-01

    Full Text Available It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of two MAP genes (MAP2121c and MAP3733c can enhance the heterologous expression of two antigens (MMP and MptD respectively, analogous to the form to which they are produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants were preserved in synthetic MMP proteins as shown by monoclonal antibody mediated ELISA. Moreover, MMP is a membrane protein in MAP, which is also targeted to the cellular surface of recombinant L. salivarius at levels comparable to MAP. Additionally, codon optimised MptD displayed the tendency to associate with the cytoplasmic membrane boundary under confocal microscopy and the intracellularly accumulated protein selectively adhered with the MptD-specific bacteriophage fMptD. This work demonstrates there is potential for L. salivarius as a viable antigen delivery vehicle for MAP, which may provide an effective mucosal vaccine against Johne’s disease.

  19. Molecular Basis Underlying Leaf Variegation of a Moth Orchid Mutant (Phalaenopsis aphrodite subsp. formosana

    Directory of Open Access Journals (Sweden)

    Chi-Chu Tsai

    2017-07-01

    Full Text Available Leaf variegation is often the focus of plant breeding. Here, we studied a variegated mutant of Phalaenopsis aphrodite subsp. formosana, which is usually used as a parent of horticultural breeding, to understand its anatomic and genetic regulatory mechanisms in variegation. Chloroplasts with well-organized thylakoids and starch grains were found only in the mesophyll cells of green sectors but not of yellow sectors, confirming that the variegation belongs to the chlorophyll type. The two-dimensional electrophoresis and LC/MS/MS also reveal differential expressions of PsbP and PsbO between the green and yellow leaf sectors. Full-length cDNA sequencing revealed that mutant transcripts were caused by intron retention. When conditioning on the total RNA expression, we found that the functional transcript of PsbO and mutant transcript of PsbP are higher expressed in the yellow sector than in the green sector, suggesting that the post-transcriptional regulation of PsbO and PsbP differentiates the performance between green and yellow sectors. Because PsbP plays an important role in the stability of thylakoid folding, we suggest that the negative regulation of PsbP may inhibit thylakoid development in the yellow sectors. This causes chlorophyll deficiency in the yellow sectors and results in leaf variegation. We also provide evidence of the link of virus CymMV and the formation of variegation according to the differential expression of CymMV between green and yellow sectors.

  20. Cytotoxicity analysis of three Bacillus thuringiensis subsp. israelensis δ-endotoxins towards insect and mammalian cells.

    Directory of Open Access Journals (Sweden)

    Roberto Franco Teixeira Corrêa

    Full Text Available Three members of the δ-endotoxin group of toxins expressed by Bacillus thuringiensis subsp. israelensis, Cyt2Ba, Cry4Aa and Cry11A, were individually expressed in recombinant acrystalliferous B. thuringiensis strains for in vitro evaluation of their toxic activities against insect and mammalian cell lines. Both Cry4Aa and Cry11A toxins, activated with either trypsin or Spodoptera frugiperda gastric juice (GJ, resulted in different cleavage patterns for the activated toxins as seen by SDS-PAGE. The GJ-processed proteins were not cytotoxic to insect cell cultures. On the other hand, the combination of the trypsin-activated Cry4Aa and Cry11A toxins yielded the highest levels of cytotoxicity to all insect cells tested. The combination of activated Cyt2Ba and Cry11A also showed higher toxic activity than that of toxins activated individually. When activated Cry4Aa, Cry11A and Cyt2Ba were used simultaneously in the same assay a decrease in toxic activity was observed in all insect cells tested. No toxic effect was observed for the trypsin-activated Cry toxins in mammalian cells, but activated Cyt2Ba was toxic to human breast cancer cells (MCF-7 when tested at 20 µg/mL.

  1. Red brome (Bromus rubens subsp. madritensis) in North America: Possible modes for early introductions, subsequent spread

    Science.gov (United States)

    Salo, L.F.

    2005-01-01

    Although invasions by exotic plants have increased dramatically as human travel and commerce have increased, few have been comprehensively described. Understanding the patterns of invasive species spread over space and time will help guide management activities and policy. Tracing the earliest appearances of an exotic plant reveals likely sites of introduction, paving the way for genetic studies to quantify founder events and identify potential source populations. Red brome (Bromus madritensis subsp. rubens) is a Mediterranean winter annual grass that has invaded even relatively undisturbed areas of western North America, where it threatens native plant communities. This study used herbarium records and contemporary published accounts to trace the early introductions and subsequent spread of red brome in western North America. The results challenge the most frequently cited sources describing the early history of this grass and suggest three possible modes for early introductions: the California Gold Rush and Central Valley wheat, southern California shipping, and northern California sheep. Subsequent periods of most rapid spread into new areas, from 1930 to 1942, and of greatest spread into new regions, during the past 50 years, coincide with warm Pacific Decadal Oscillation regimes, which are linked to increased winter precipitation in the southwestern USA and northern Mexico. Global environmental change, including increased atmospheric CO2 levels and N deposition, may be contributing to the success of red brome, relative to native species.

  2. Oligosaccharide binding proteins from Bifidobacterium longum subsp. infantis reveal a preference for host glycans.

    Directory of Open Access Journals (Sweden)

    Daniel Garrido

    2011-03-01

    Full Text Available Bifidobacterium longum subsp. infantis (B. infantis is a common member of the infant intestinal microbiota, and it has been characterized by its foraging capacity for human milk oligosaccharides (HMO. Its genome sequence revealed an overabundance of the Family 1 of solute binding proteins (F1SBPs, part of ABC transporters and associated with the import of oligosaccharides. In this study we have used the Mammalian Glycan Array to determine the specific affinities of these proteins. This was correlated with binding protein expression induced by different prebiotics including HMO. Half of the F1SBPs in B. infantis were determined to bind mammalian oligosaccharides. Their affinities included different blood group structures and mucin oligosaccharides. Related to HMO, other proteins were specific for oligomers of lacto-N-biose (LNB and polylactosamines with different degrees of fucosylation. Growth on HMO induced the expression of specific binding proteins that import HMO isomers, but also bind blood group and mucin oligosaccharides, suggesting coregulated transport mechanisms. The prebiotic inulin induced other family 1 binding proteins with affinity for intestinal glycans. Most of the host glycan F1SBPs in B. infantis do not have homologs in other bifidobacteria. Finally, some of these proteins were found to be adherent to intestinal epithelial cells in vitro. In conclusion, this study represents further evidence for the particular adaptations of B. infantis to the infant gut environment, and helps to understand the molecular mechanisms involved in this process.

  3. Inferring biomarkers for Mycobacterium avium subsp. paratuberculosis infection and disease progression in cattle using experimental data

    Science.gov (United States)

    Magombedze, Gesham; Shiri, Tinevimbo; Eda, Shigetoshi; Stabel, Judy R.

    2017-03-01

    Available diagnostic assays for Mycobacterium avium subsp. paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN-γ, ELISA-antibody and fecal shedding experimental sensitivity scores for MAP infection detection and disease progression. We used both statistical methods and dynamic mathematical models to (i) evaluate the empirical assays (ii) infer and explain biological mechanisms that affect the time evolution of the biomarkers, and (iii) predict disease stages of 57 animals that were naturally infected with MAP. This analysis confirms that the fecal test is the best marker for disease progression and illustrates that Th1/Th2 (IFN-γ/ELISA antibodies) assays are important for infection detection, but cannot reliably predict persistent infections. Our results show that the theoretical simulated macrophage-based assay is a potential good diagnostic marker for MAP persistent infections and predictor of disease specific stages. We therefore recommend specifically designed experiments to test the use of a based assay in the diagnosis of MAP infections.

  4. Ectopic accumulation of linalool confers resistance to Xanthomonas citri subsp. citri in transgenic sweet orange plants.

    Science.gov (United States)

    Shimada, Takehiko; Endo, Tomoko; Rodríguez, Ana; Fujii, Hiroshi; Goto, Shingo; Matsuura, Takakazu; Hojo, Yuko; Ikeda, Yoko; Mori, Izumi C; Fujikawa, Takashi; Peña, Leandro; Omura, Mitsuo

    2017-05-01

    In order to clarify whether high linalool content in citrus leaves alone induces strong field resistance to citrus canker caused by Xanthomonas citri subsp. citri (Xcc), and to assess whether this trait can be transferred to a citrus type highly sensitive to the bacterium, transgenic 'Hamlin' sweet orange (Citrus sinensis L. Osbeck) plants over-expressing a linalool synthase gene (CuSTS3-1) were generated. Transgenic lines (LIL) with the highest linalool content showed strong resistance to citrus canker when spray inoculated with the bacterium. In LIL plants inoculated by wounding (multiple-needle inoculation), the linalool level was correlated with the repression of the bacterial titer and up-regulation of defense-related genes. The exogenous application of salicylic acid, methyl jasmonate or linalool triggered responses similar to those constitutively induced in LIL plants. The linalool content in Ponkan mandarin leaves was significantly higher than that of leaves from six other representative citrus genotypes with different susceptibilities to Xcc. We propose that linalool-mediated resistance might be unique to citrus tissues accumulating large amounts of volatile organic compounds in oil cells. Linalool might act not only as a direct antibacterial agent, but also as a signal molecule involved in triggering a non-host resistance response against Xcc. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  5. The type III protein secretion system contributes to Xanthomonas citri subsp. citri biofilm formation

    KAUST Repository

    Zimaro, Tamara

    2014-04-18

    Background: Several bacterial plant pathogens colonize their hosts through the secretion of effector proteins by a Type III protein secretion system (T3SS). The role of T3SS in bacterial pathogenesis is well established but whether this system is involved in multicellular processes, such as bacterial biofilm formation has not been elucidated. Here, the phytopathogen Xanthomonas citri subsp. citri (X. citri) was used as a model to gain further insights about the role of the T3SS in biofilm formation. Results: The capacity of biofilm formation of different X. citri T3SS mutants was compared to the wild type strain and it was observed that this secretion system was necessary for this process. Moreover, the T3SS mutants adhered proficiently to leaf surfaces but were impaired in leaf-associated growth. A proteomic study of biofilm cells showed that the lack of the T3SS causes changes in the expression of proteins involved in metabolic processes, energy generation, exopolysaccharide (EPS) production and bacterial motility as well as outer membrane proteins. Furthermore, EPS production and bacterial motility were also altered in the T3SS mutants. Conclusions: Our results indicate a novel role for T3SS in X. citri in the modulation of biofilm formation. Since this process increases X. citri virulence, this study reveals new functions of T3SS in pathogenesis. 2014 Zimaro et al.; licensee BioMed Central Ltd.

  6. Reducing the Bitterness of Tuna (Euthynnus pelamis Dark Meat with Lactobacillus casei subsp. casei ATCC 393

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    Ernani S. Sant’Anna

    2004-01-01

    Full Text Available During the process of canning tuna fish, considerable amounts of dark tuna meat are left over because of its bitterness, which are then used in the production of animal food. Fermentation with Lactobacillus casei subsp. casei ATCC 393 was used as an alternative to reduce this bitter taste. Samples of meat were prepared, vacuum packed and then stored at –18 °C. The frozen dark meat was used immediately after defrosting and the experiment was carried out with 2 and 4 % of NaCl with the addition of 2 and 4 % of glucose, respectively. The dark tuna meat was inoculated with lactic acid bacteria (LAB and fermented at 10 °C for 30 days. The fermentation process was monitored through bacteriological and chemical analyses, when an increase of acidity and the corresponding decrease of pH were observed due to the prevalence of LAB. Sensorial analysis, using a test of multiple comparison, was carried out with pastes of fermented dark tuna meat and presented a significant difference when compared to the paste control, indicating the reduction of bitter taste.

  7. The Effect of Citrus Essential Oils and Their Constituents on Growth of Xanthomonas citri subsp. citri.

    Science.gov (United States)

    Mirzaei-Najafgholi, Hossein; Tarighi, Saeed; Golmohammadi, Morteza; Taheri, Parissa

    2017-04-14

    Citrus bacterial canker (CBC) caused by Xanthomonas citri subsp. citri ( Xcc ), is the most devastating of the citrus diseases worldwide. During our study, we found that Essential oils (EOs) of some citrus cultivars are effective on Xcc . Therefore, it prompted us to determine the plant metabolites responsible for the antibacterial properties. We obtained EOs from some locally cultivated citrus by using a Clevenger apparatus and their major constituents were identified by gas chromatography/mass spectrometry (GC-MS). The effect of Citrus aurantium , C. aurantifolia , Fortunella sp. EOs and their major constituents were evaluated against Xcc -KVXCC1 using a disk diffusion assay. Minimal inhibitory and bactericidal concentration of the EOs and their constituents were determined using the broth microdilution method. C. aurantium , C. aurantifolia Eos, and their major constituents including citral, linalool, citronellal, geraniol, α-terpineol, and linalyl acetate indicated antibacterial effects against Xcc . The C. aurantifolia EO and citral showed the highest antibacterial activity among the tested EOs and constituents with inhibition zones of 15 ± 0.33 mm and 16.67 ± 0.88 mm, respectively. Synergistic effects of the constituents were observed between α-terpineol-citral, citral-citronellal, citral-geraniol, and citronellal-geraniol by using a microdilution checkerboard assay. Transmission electron microscopy revealed that exposure of Xcc cells to citral caused cell wall damage and altered cytoplasmic density. We introduced C. aurantifolia and C. aurantium EOs, and their constituents citral, α-terpineol, citronellal, geraniol, and linalool as possible control agents for CBC.

  8. Mycobacterium abscessus subsp. abscessus Lung Disease: Drug Susceptibility Testing in Sputum Culture Negative Conversion

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    Takehiko Kobayashi

    2018-01-01

    Full Text Available Background: Among Mycobacterium abscessus complex infections, patients with M. abscessus subsp. abscessus (MAA lung disease are difficult to treat and no standard therapy has been established. Few reports have investigated the drug susceptibility of these strains. We retrospectively investigated how in vitro drug susceptibility testing (DST of MAA affects the induction of sputum conversion using pharmacotherapy. Methods: Patients with MAA lung disease diagnosed and treated between 2010 and 2014 at our hospital were enrolled and divided into Group A (sputum conversion without relapse within 1 year and Group B (persistent positive cultured or negative conversion with relapse. MAA was identified in M. abscessus using sequence with genotyping, and DST of MAA was performed. Results: We assessed 23 patients (9 males and 14 females. There were 8 patients in Group A and 15 in Group B. Higher prevalence of susceptible isolates for clarithromycin (CAM susceptibility on day 14 was noted in Group A than in Group B (P = 0.03 and no significant difference observed in the two groups for other drugs. Conclusions: In vitro DST of MAA, especially CAM susceptibility on day 14, affected the results of negative conversion. No other drugs were found to affect sputum culture negative conversion.

  9. The effect of nisin from Lactococcus lactis subsp. lactis on refrigerated patin fillet quality

    Science.gov (United States)

    Adilla, S. N.; Utami, R.; Nursiwi, A.; Nurhartadi, E.

    2017-04-01

    The effect of nisin from Lactococcus lactis subsp. lactis with spraying method application on quality of patin fillet during refrigerated storage (4±1°C) was investigated. The quality of patin fillet based on total plate count (TPC), pH, TVB-N, and TBA values during 16 days at 4±1°C. Completely Randomized Design (CDR) was used in one factor (nisin activity) at 0 IU/ml, 500 IU/ml, 1000 IU/ml, and 2000 IU/ml. The observation was done at 0, 4th, 8th, 12th, and 16th days of storage. The result showed that variation of nisin activity significantly affected the quality of fillet according to TPC, pH, and TVB-N values, however no significant difference on the obtained of TBA value. Nisin in 500 IU/ml, 1000 IU/ml, and 2000 IU/ml could extend the shelf-life of fillet until 4th, 8th, and 12th days respectively based on standard in all parameters.

  10. Genetic transformation of intact Lactococcus lactis subsp. lactis by high-voltage electroporation

    Energy Technology Data Exchange (ETDEWEB)

    McIntyre, D.A.; Harlander, S.K. (Univ. of Minnesota, St. Paul (USA))

    1989-03-01

    The objective of this study was to develop a system for electroporating intact cells of Lactococcus lactis subsp. lactis LM0230 (previously designated Streptococcus lactis LM0230) with a commercially available electroporation unit. Parameters which influenced the efficiency of transformation included growth phase and final concentration of cells, ionic strength of the suspending medium, concentration of plasmid DNA, and the amplitude and duration of the pulse. Washed suspensions of intact cells suspended in deionized distilled water were subjected to one high-voltage electric pulse varying in voltage (300 to 900 V corresponding to field strengths of 5 to 17 kV/cm) and duration (100 {mu}s to 1 s). Transformation efficiencies of 10{sup 3} transformants per {mu}g of DNA were obtained when dense suspensions (final concentration, 5 {times} 10{sup 10} CFU/ml) of stationary-phase cells were subjected to one pulse with a peak voltage of 900 V (field strength, 17 kV/cm) and a pulse duration of 5 ms in the presence of plasmid DNA. Dilution of porated cells in broth medium followed by an expression period of 2 h at 30{degree}C was beneficial in enhancing transformation efficiencies. Plasmids ranging in size from 9.8 to 30.0 kilobase pairs could be transformed by this procedure.

  11. Predictor variables for and complications associated with Streptococcus equi subsp equi infection in horses.

    Science.gov (United States)

    Duffee, Lauren R; Stefanovski, Darko; Boston, Raymond C; Boyle, Ashley G

    2015-11-15

    To evaluate predictor variables for and complications associated with Streptococcus equi subsp equi infection (strangles) in horses. Retrospective case-control study. 108 horses with strangles (cases) and 215 horses without strangles (controls). Medical records from January 2005 through July 2012 were reviewed. Cases were defined as horses with clinical signs of strangles (pyrexia, retropharyngeal lymphadenopathy, and mucopurulent nasal discharge) that were associated with a confirmed strangles outbreak or had positive results for S equi on PCR assay or bacteriologic culture. Controls were defined as horses with pyrexia that did not meet the other criteria for cases. Data compared between cases and controls included signalment, clinical signs, diagnostic test results, and disease complications and outcome. Logistic regression was used to identify variables associated with strangles and its complications. Clinical signs of strangles were not evident in 12 of 25 cases classified as S equi carriers (infected > 40 days). Predictor variables associated with strangles included mucopurulent nasal discharge and external abscesses in the pharyngeal region. Strangles was more likely to be diagnosed in the spring than in the summer. Cases with anemia were more likely to develop purpura hemorrhagica than were cases without anemia. No risk factors were identified for the development of guttural pouch empyema or metastatic abscesses. Results indicated that not all horses infected with S equi develop clinical signs of strangles. We recommend that guttural pouch endoscopy and lavage with PCR assay of lavage fluid samples be performed to identify S equi carrier horses.

  12. Streptococcus equi subsp. zooepidemicus isolates from equine infectious endometritis belong to a distinct genetic group.

    Science.gov (United States)

    Rasmussen, Camilla Dooleweerdt; Haugaard, Maria Mathilde; Petersen, Morten Roenn; Nielsen, Jesper Møller; Pedersen, Hanne Gervi; Bojesen, Anders Miki

    2013-04-18

    Streptococcus equi subsp. zooepidemicus is the pathogen most commonly isolated from the uterus of mares. S. zooepidemicus is an opportunistic pathogen and part of the resident flora in the caudal reproductive tract. The aim of this study was to investigate whether a genotypically distinct subpopulation of S. zooepidemicus is associated with endometritis in the mare, by genotyping and comparing uterine S. zooepidemicus strains with isolates from the vagina and clitoral fossa. Mares with (n=18) or without (n=11) clinical symptoms of endometritis were included. Uterine samples were obtained using a guarded endometrial biopsy punch, whereas a swab was used to recover samples from the cranial vagina and the clitoral fossa. If S. zooepidemicus was present, up to three colonies were selected from each anatomical location (max. 9 isolates per mare). Bacterial isolates were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). S. zooepidemicus was isolated from the endometrium of 12 mares. A total of 88 isolates were analyzed by PFGE: 31 from the endometrium, 26 from the cranial vagina and 31 isolates from the clitoral fossa. For MLST 21 isolates were chosen. Results demonstrated a higher genetic similarity of the isolates obtained from infectious endometritis compared to isolates obtained from the caudal reproductive tract. In conclusion, we demonstrate for the first time that a genetically distinct group of S. zooepidemicus is associated with infectious endometritis in the mare.

  13. Evaluation of a commercially available modified-live Streptococcus equi subsp equi vaccine in ponies.

    Science.gov (United States)

    Borst, Luke B; Patterson, Sheila K; Lanka, Saraswathi; Barger, Anne M; Fredrickson, Richard L; Maddox, Carol W

    2011-08-01

    To evaluate a commercially available modified-live Streptococcus equi subsp equi vaccine for safety and persistence in vaccinated ponies and to detect recombination or reversion events in the vaccine strain. 5 ponies that were 1.5 to 8 years old (group 1) and 4 ponies that were 6 months old (group 2). Ponies were vaccinated, with a subsequent booster vaccination 2 to 3 weeks later, and monitored for 50 days. At booster vaccination, an equal amount of a tetracycline-resistant wild-type strain of S equiwas administered. Recovery of all strains was performed by use of bacteriologic culture and PCR assays. Ponies in group 1 had background antibody titers against S equi antigen before vaccination despite the lack of known exposure to S equi. Ponies in group 2 were immunologically naïve. Increases in anti-S equi antibody titers were detected in both groups. Ponies in group 1 did not have clinical signs of disease caused by S equi. In group 2, all ponies developed abscesses in retropharyngeal lymph nodes; 1 pony developed severe clinical disease and was euthanized. The vaccine strain was recovered from ponies in group 2 for up to 24 days after vaccination. Although the vaccine was successful in inducing IgG antibodies against S equi in all ponies, findings suggested that the vaccine may have caused substantial morbidity and some deaths in the young ponies. In young ponies, the vaccine strain persisted in tissues for weeks; however, no evidence of recombination was detected.

  14. [Antimicrobial susceptibility in isolates of Streptococcus equi subsp. equi from Buenos Aires province, Argentina].

    Science.gov (United States)

    Bustos, Carla P; Marfil, María J; Lanza, Natalia S; Guida, Nora

    2017-11-30

    Streptococcus equi subsp. equi is the etiologic agent of strangles, an infectious disease affecting the upper respiratory tract and head and neck lymph nodes of equines. Routine antimicrobial therapy includes penicillin (PEN) as antibiotic of first choice. Streptococci are usually susceptible to PEN and only a few antimicrobial studies had been performed. The aim of this work was to study the antimicrobial susceptibility profile of S. equi from Buenos Aires, Argentina. Ninety-two isolates were studied by the single disk method to PEN, cefotaxime, erythromycin (ERY), tetracycline, enrofloxacin (ENR), trimethoprim sulfamethoxazole (TMS), ciprofloxacin, clindamycin (CLI), streptomycin (STR) and florfenicol. Minimum inhibitory concentration (MIC) to PEN and antibiotics with resistance and intermediate susceptibility were tested. High percentages of susceptibility were obtained by the disk diffusion method and MIC values of PEN, TMS and CLI were found to be under the breakpoint values. Resistant strains of ENR and STR with MIC 50 , MIC 90 and MIC ranges above breakpoints were identified. These findings confirm that PEN may be used empirically because resistant strains were not found in Buenos Aires. Emphasis is placed on the rational use of antibiotics to achieve therapeutic success, to prevent chronicity, recurrence of infections and the emergence of resistance. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  15. Homology Modeling and Functional Characterization of PR-1a Protein of Hordeum vulgare subsp. Vulgare.

    Science.gov (United States)

    Aslanzadeh, Vahid; Ghaderian, Mostafa

    2012-01-01

    Pathogenesis-related protein 1a of Hordeum vulgare subsp. Vulgare (HvPR-1a) is induced by various pathogens and stress related factors. It plays important roles in plant defense system. Since the discovery of HvPR-1a a great deal of research has been focused on its isolation and characterization. However, three dimensional structure of HvPR-1a is still unknown. 3D structure can be used for determining protein function, and identifying novel protein folds and potential targets for regulation. The protein model was developed using MODELLER 9v10. Physicochemical characterization and functional annotation of the model carried out with Expasy's ProtParam server and three different conserved domain finding programs including InterProScan, Proteins Families Database (Pfam), and NCBI Conserved Domains Database (NCBI-CDD). Applying validation programs revealed that the model has good quality and the RMSD value is 0.7. The predicted model submitted in Protein Model Database, PMDB for public use. This model will be used in wide range of studies for functional analysis and improvement activity of the protein.

  16. Preventive effect of taraxasteryl acetate from Inula britannica subsp. japonica on experimental hepatitis in vivo.

    Science.gov (United States)

    Iijima, K; Kiyohara, H; Tanaka, M; Matsumoto, T; Cyong, J C; Yamada, H

    1995-02-01

    The survival rate for acute hepatic failure induced by Propionibacterium acnes and lipopolysaccharide (LPS) was increased when a hot water extract from the flowers of Inula britannica L. subsp. japonica Kitam. was injected into the experimental hepatitis mice, and anti-hepatitis substances could be extracted with CHCl3. The CHCl3 extract from I.britannica was fractionated and anti-hepatitis fractions IB-3-2 and IB-3-3 were obtained. IB-3-3 had the most potent anti-hepatitis activity among the fractions but further purification of the active compound was not achieved because of the low yield. IB-3-2 contained only one substance which was identified to be taraxasteryl acetate by 1H- and 13C-NMR and MS. Taraxasteryl acetate showed potent preventive activity against acute hepatic failure induced by P.acnes and LPS in a dose-dependent manner, however deacetylation and modification of the olefinic bonds significantly decreased the anti-hepatitis activity of taraxasteryl acetate. Taraxasteryl acetate also inhibited the increment of plasma transaminase on acute hepatic failure induced by carbon tetrachloride (CCl4) or D-galactosamine. From a histological study it appeared that degeneration and necrosis, which were observed in the liver from CCl4 mice, were not found in the liver cells from taraxasteryl acetate treated mice. These results indicates that taraxasteryl acetate shows preventive effects on experimental hepatitis caused by either immunologically induced injuries or hepatotoxic chemicals.

  17. Comparison of rapid diagnostic tests to detect Mycobacterium avium subsp. paratuberculosis disseminated infection in bovine liver.

    Science.gov (United States)

    Zarei, Mehdi; Ghorbanpour, Masoud; Tajbakhsh, Samaneh; Mosavari, Nader

    2017-08-01

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease, a chronic enteritis in cattle and other domestic and wild ruminants. The presence of MAP in tissues other than intestines and associated lymph nodes, such as meat and liver, is a potential public health concern. In the present study, the relationship between the results of rapid diagnostic tests of the Johne's disease, such as serum ELISA, rectal scraping PCR, and acid-fast staining, and the presence of MAP in liver was evaluated. Blood, liver, and rectal scraping samples were collected from 200 slaughtered cattle with unknown Johne's disease status. ELISA was performed to determine the MAP antibody activity in the serum. Acid-fast staining was performed on rectal scraping samples, and PCR was performed on rectal scraping and liver samples. PCR-positive liver samples were used for mycobacterial culture. Overall, the results of this study demonstrated that MAP can be detected and cultured from liver of slaughtered cattle and rapid diagnostic tests of Johne's disease have limited value in detecting cattle with MAP infection in liver. These findings show that the presence of MAP in liver tissue may occur in cows with negative results for rapid diagnostic tests and vice versa. Hence, liver might represent another possible risk of human exposure to MAP. Given concerns about a potential zoonotic role for MAP, these results show the necessity to find new methods for detecting cattle with MAP disseminated infection.

  18. Biosorption of copper(II) by Marrubium globosum subsp. globosum leaves powder: effect of chemical pretreatment.

    Science.gov (United States)

    Yazici, Hüseyin; Kiliç, Mehmet; Solak, Murat

    2008-03-01

    The study was aimed at determining the effect of chemical pretreatment on copper(II) biosorption by Marrubium globosum subsp. globosum leaves. The uptake capacity of the biomass was increased by chemical pretreatment when compared with the raw biomass. The results of biosorption experiments, carried out at the conditions of 50 mg l(-1) initial metal concentration and pH 5.5, showed that pretreating the biomass with alkali solutions (laundry detergent, sodium hydroxide and sodium bicarbonate, 0.5 M) improved the biosorption capacity of biomass (45.90, 45.78 and 43.91%, respectively) compared with raw biomass. Pretreatment with sulfuric and nitric acid solutions, 0.5 M, increased the biosorption capacity of biomass by 11.82 and 10.18%, respectively, while there was no considerable change in the biosorption capacity of biomass (0.35%) after pretreatment with formic acid solution, 0.5 M. Furthermore, sodium chloride and calcium chloride, 0.5 M, pretreatments resulted in the improvement in biosorption capacity of biomass (31.38 and 26.69%, respectively). FT-IR analysis revealed that hydroxyl and carboxyl functional groups were mainly responsible for copper(II) biosorption.

  19. Pectobacterium carotovorum subsp. carotovorum – the causal agent of broccoli soft rot in Serbia

    Directory of Open Access Journals (Sweden)

    Katarina Gašić

    2014-12-01

    Full Text Available Soft rot symptoms were observed on broccoli plants in several commercial fields in the western part of Serbia. Six strains of bacteria were isolated from diseased tissues and identified as Pectobacterium carotovorum subsp. carotovorum using conventional bacteriological and molecular methods. All strains were non-fluorescent, gram-negative, facultative anaerobes, oxidase-negative and catalase-positive, causing soft rot on potato and carrot slices and did not induce hypersensitive reaction on tobacco leaves. They grew in 5% NaCl and at 37°C, did not produce acid from α-methyl glucoside, sorbitol and maltose, nor reducing substances from sucrose, but utilized lactose and trehalose, and did not produce indole or lecithinase. The investigated strains showed characteristic growth on Logan’s medium and did not produce blue pigmented indigoidine on GYCA medium nor “fried egg” colonies on PDA. The identity of strains was confirmed by ITS-PCR and ITS-RFLP analyses and by sequence analysis of the 16S rRNA gene. In a pathogenicity assay, all strains caused tissue discoloration and soft rot development on inoculated broccoli head tissue fragments.

  20. Screening bactericidal effect of Pectobacterium carotovorum subsp. carotovorum strains against causal agent of potato soft rot.

    Science.gov (United States)

    Kazemi-Zaromi, Samaneh; Baghaee-Ravari, Sareh; Khodaygan, Pejman; Falahati-Rastegar, Mahrokh

    2016-02-01

    This study focuses on the potential of Pectobacterium carotovorum subsp. carotovorum (Pcc) strains producing bacteriocin as a tool to control potato soft rot disease. Thirty out of 48 purified bacterial strains were characterized as Pcc using specific PCR and phenotypic tests. The pathogenicity and pectate degrading assays were recorded positive for 13 strains. Bacteriocin typing clustered producers into three groups according to their antimicrobial spectra. Majority of the producers except strains of group II showed antibacterial activity toward relative genus and the role of UV or mitomycin C was inductive. In addition, none of the distant genus was sensitive to Pcc bacteriocins except Rhizobium vitis. Molecular detection of four bacteriocins including carotovoricin, carosin S1, S2 and carosin D was performed. Overall, 54.5% of group I, 47.3 and 70% of groups II and III strains carried carotovoricin and four strains harbored gene corresponding to carosin S1. According to our data divers antimicrobial patterns obtained by Pcc strains and existence of new bateriocines could be possible. Moreover, our findings recommended that direct application of P29 or expression of corresponding genes of Pog22 or P21 in a nonpathogenic strain as a biocontrol agent may improve soft rot disease control. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Lettuce genotype resistance to "soft rot" caused by Pectobacterium carotovorum subsp. carotovorum

    Directory of Open Access Journals (Sweden)

    Kátia Cilene da Silva Felix

    2014-08-01

    Full Text Available Soft rot, caused by Pectobacterium carotovorum subsp. carotovorum (Pcc, is the main bacterial disease affecting lettuce (Lactuca sativa L. crops in Brazil and leads to significant yield losses. This study aimed to assess the reaction of lettuce genotypes to soft rot induced by a virulent isolate and the stability of the resistance to three isolates varying in virulence. Using a descriptive ordinal scale ranging from 1 to 9 a classification system was defined: class 1 = resistant (R: severity (Sev 3.5. Of the 41 tested genotypes, 14 were classified as MR and 27 as S when inoculated with a Pcc isolate of intermediate virulence. Eleven of these genotypes (four S and seven MR were selected to test their resistance stability against three other isolates with an increasing degree of virulence (Pcc36 < Pcc-A1.1 < Pcc-23. Out of the 11 genotypes eight retained the original classification and three moved from S to MR resistant class when challenged with the least virulent isolate. Vitória de Santo Antão was the only genotype classified as MR for all tested isolates and is a promising candidate for durable soft rot resistance breeding.

  2. Propionic acid production in glycerol/glucose co-fermentation by Propionibacterium freudenreichii subsp. shermanii.

    Science.gov (United States)

    Wang, Zhongqiang; Yang, Shang-Tian

    2013-06-01

    Propionibacterium freudenreichii subsp. shermanii can ferment glucose and glycerol to propionic acid with acetic and succinic acids as two by-products. Propionic acid production from glucose was relatively fast (0.19 g/Lh) but gave low product yield (~0.39 g/g) and selectivity (P/A: ~2.6; P/S: ~4.8). In contrast, glycerol with a more reduced state gave a high propionic acid yield (~0.65 g/g) and selectivity (P/A: ~31; P/S: ~11) but low productivity (0.11 g/L h). On the other hand, co-fermentation of glycerol and glucose at an appropriate mass ratio gave both a high yield (0.54-0.65 g/g) and productivity (0.18-0.23 g/L h) with high product selectivity (P/A: ~14; P/S: ~10). The carbon flux distributions in the co-fermentation as affected by the ratio of glycerol/glucose were investigated. Finally, co-fermentation with cassava bagasse hydrolysate and crude glycerol in a fibrous-bed bioreactor was demonstrated, providing an efficient way for economic production of bio-based propionic acid. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Interactive behavior of Saccharomyces cerevisiae, Bacillus pumilus and Propionibacterium freudenreichii subsp. shermanii.

    Science.gov (United States)

    Odame-Darkwah, J K; Marshall, D L

    1993-09-01

    Prevention of ropy bread caused by mucoid variants of certain bacilli presents a major problem for developing countries where cost of preservatives is prohibitive. Control of ropiness may be achieved by using propionic acid-producing bacteria in mixed culture with leavening yeasts. Therefore, interaction studies between Propionibacterium freudenreichii subsp. shermanii, Bacillus pumilus and Saccharomyces cerevisiae were conducted in a chemically defined medium to test the relevance of such an approach. Growth of vegetative cells and germination of spores of B. pumilus were inhibited in media preincubated with P. shermanii at 30 degrees C for 13 h. Inhibition was bacteriostatic for the first 6 h of incubation, becoming bactericidal between 6 and 12 h. Inhibition of B. pumilus spore germination was greater than inhibition of growth of vegetative cells of the bacterium. Culturing of either P. shermanii with S. cerevisiae or B. pumilus with S. cerevisiae did not produce inhibitory effects on any of the organisms. Inhibition of B. pumilus by P. shermanii may be useful for prevention of ropiness in bread prepared by the sponge method, involving fermentation of a portion of the dough.

  4. [Isolation and purification of a protective protein from Propionibacterium freudenreichii subsp. Shermanii].

    Science.gov (United States)

    Zinchenko, A A; Vorob'eva, L I; Gordeeva, E A; Khodzhaev, E Iu; Ponomareva, G M; Nokel', E A

    1998-01-01

    A protein responsible for the protective and reactivating activities of two active fractions (AF1 and AF2) of the cells of Propionibacterium freudenreichii subsp. shermanii was isolated. The active fraction AF1 was obtained by fractional precipitation of the cell-free extract of propionic acid bacteria between 20 and 40% ammonium sulfate saturation, whereas fraction AF2 was precipitated between 60 and 80% saturation. Further fractionation of AF1 and AF2 by gel filtration on Sephacryl S-200 and by ion-exchange chromatography on DEAE-Sepharose yielded seven active subfractions, as revealed by testing for their protective activity on UV-inactivated cells of Escherichia coli. Analysis of subfraction AF2-2.5 by SDS-electrophoresis and HPLC showed that it contained an apparently homogeneous protein with a molecular mass of 44 +/- 2 kDa. The concentrational dependence of the protective activity of this protein was derived. Peptides of subfractions AF2-2.1 and AF2-2.2 with molecular masses lower than 15 kDa also exhibited protective activity.

  5. Detection and Characterization of Histamine-Producing Strains of Photobacterium damselae subsp. damselae Isolated from Mullets

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    Marcello Trevisani

    2017-06-01

    Full Text Available Photobacterium damselae subsp. damselae (Pdd is considered to be an emerging pathogen of marine fish and has also been implicated in cases of histamine food poisoning. In this study, eight strains isolated from mullets of the genera Mugil and Liza captured in the Ligurian Sea were characterized, and a method to detect histamine-producing Pdd from fish samples was developed. The histamine-producing potential of the strains was evaluated in culture media (TSB+ using a histamine biosensor. Subsequently, two strains were used to contaminate mackerel fillets (4 or 40 CFU/g, simulating a cross-contamination on the selling fish stalls. Sample homogenates were enriched in TSB+. The cultures were then inoculated on thiosulfate-citrate-bile salts-sucrose agar (TCBS and the dark green colonies were cultured on Niven agar. The violet isolates were characterized using specific biochemical and PCR based tests. All Pdd strains were histamine producers, yielding concentration varying from 167 and 8977 µg/mL in TSB+ cultures incubated at 30 °C for 24 h. Pdd colonies were detected from the inoculated mackerel samples and their histidine decarboxylase gene was amplified using species-specific primer pairs designed for this study. The results indicate that mullets can be source of Pdd and the fish retailers needs to evaluate the risk posed by cross-contamination on the selling fish stalls.

  6. Outbreak of upper respiratory disease in horses caused by Streptococcus equi subsp. zooepidemicus ST-24.

    Science.gov (United States)

    Lindahl, Susanne B; Aspán, Anna; Båverud, Viveca; Paillot, Romain; Pringle, John; Rash, Nicola L; Söderlund, Robert; Waller, Andrew S

    2013-09-27

    Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is generally considered a commensal and an opportunistic pathogen of the upper airways in horses. Establishing whether certain strains of S. zooepidemicus can cause upper respiratory disease as a host-specific pathogen of horses, and if there are certain genogroups of S. zooepidemicus that are more virulent than others is of major clinical importance. In this study, we describe an outbreak of upper respiratory disease in horses that was associated with S. zooepidemicus. Upper respiratory samples were cultured, analyzed by real-time PCR for S. zooepidemicus and S. equi, and genetically differentiated by sequencing of the SzP protein gene and multi-locus sequence typing (MLST). Serum samples were analyzed for antibodies against S. equi and common viral respiratory pathogens. The ST-24 strain of S. zooepidemicus was isolated from all horses with clinical signs of disease, while the healthy horses carried other strains of S. zooepidemicus. Bacteriological, molecular and serological analyses strongly suggest that a single strain (ST-24) was responsible for the disease outbreak, and that certain strains of this presumed commensal may be more virulent than others. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. From mouth to macrophage: mechanisms of innate immune subversion by Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Arsenault, Ryan J; Maattanen, Pekka; Daigle, Joanna; Potter, Andrew; Griebel, Philip; Napper, Scott

    2014-05-15

    Johne's disease (JD) is a chronic enteric infection of cattle caused by Mycobacterium avium subsp. paratuberculosis (MAP). The high economic cost and potential zoonotic threat of JD have driven efforts to develop tools and approaches to effectively manage this disease within livestock herds. Efforts to control JD through traditional animal management practices are complicated by MAP's ability to cause long-term environmental contamination as well as difficulties associated with diagnosis of JD in the pre-clinical stages. As such, there is particular emphasis on the development of an effective vaccine. This is a daunting challenge, in large part due to MAP's ability to subvert protective host immune responses. Accordingly, there is a priority to understand MAP's interaction with the bovine host: this may inform rational targets and approaches for therapeutic intervention. Here we review the early host defenses encountered by MAP and the strategies employed by the pathogen to avert or subvert these responses, during the critical period between ingestion and the establishment of persistent infection in macrophages.

  8. Oral Fusobacterium nucleatum subsp. polymorphum binds to human salivary α-amylase.

    Science.gov (United States)

    Zulfiqar, M; Yamaguchi, T; Sato, S; Oho, T

    2013-12-01

    Fusobacterium nucleatum acts as an intermediate between early and late colonizers in the oral cavity. In this study, we showed that F. nucleatum subsp. polymorphum can bind to a salivary component with a molecular weight of approximately 110 kDa and identified the protein and another major factor of 55 kDa, as salivary α-amylase by time-of-flight mass spectrometry and immuno-reactions. Salivary α-amylase is present in both monomeric and dimeric forms and we found that formation of the dimer depends on copper ions. The F. nucleatum adhered to both monomeric and dimeric salivary α-amylases, but the numbers of bacteria bound to the dimeric form were more than those bound to the monomeric form. The degree of adherence of F. nucleatum to four α-amylases from different sources was almost the same, however its binding to β-amylase was considerably decreased. Among four α-amylase inhibitors tested, acarbose and type 1 and 3 inhibitors derived from wheat flour showed significant activity against the adhesion of F.nucleatum to monomeric and dimeric amylases, however voglibose had little effect. Moreover F. nucleatum cells inhibited the enzymatic activity of salivary α-amylase in a dose-dependent manner. These results suggest that F. nucleatum plays more important and positive role as an early colonizer for maturation of oral microbial colonization. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  9. New Insights into the Enterococcus faecium and Streptococcus gallolyticus subsp. gallolyticus Host Interaction Mechanisms.

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    Ana María Sánchez-Díaz

    Full Text Available Enterococcus faecium and Streptococcus gallolyticus subsp. gallolyticus (S. gallolyticus were classically clustered into the Lancefield Group D streptococci and despite their taxonomic reclassification still share a similar genetic content and environment. Both species are considered as opportunistic pathogens. E. faecium is often associated with nosocomial bacteraemia, and S. gallolyticus is sporadically found in endocarditis of colorectal cancer patients. In both cases, the source of infection is commonly endogenous with a translocation process that launches through the intestinal barrier. To get new insights into the pathological processes preceding infection development of both organisms, we used an in vitro model with Caco-2 cells to study and compare the adhesion, invasion and translocation inherent abilities of 6 E. faecium and 4 S. gallolyticus well-characterized isolates. Additionally, biofilm formation on polystyrene, collagen I and IV was also explored. Overall results showed that E. faecium translocated more efficiently than S. gallolyticus, inducing a destabilization of the intestinal monolayer. Isolates Efm106, Efm121 and Efm113 (p < .001 compared to Ef222 exhibited the higher translocation ability and were able to adhere 2-3 times higher than S. gallolyticus isolates. Both species preferred the collagen IV coated surfaces to form biofilm but the S. gallolyticus structures were more compact (p = .01. These results may support a relationship between biofilm formation and vegetation establishment in S. gallolyticus endocarditis, whereas the high translocation ability of E. faecium high-risk clones might partially explain the increasing number of bacteraemia.

  10. Ciclo de vida de Pellaea ternifolia (Cav. Link subsp. ternifolia (Pteridaceae-Polypodiidae

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    Jorge Huerta-Zavala

    2015-08-01

    Full Text Available Se describe el ciclo de vida dePellaea ternifolia(Cav. Link subsp.ternifolia. Los especímenes fueron recolectados en un bosque dePinusperturbado en San Miguel de los Alcanfores, municipio de Tlaxco, Tlaxcala, México. Las esporas fueron sembradas en vasos de unicel de 262 ml con cerámica (barro molido, piedra de río, tela de mosquitero y tres soportes de cultivo (tierra, maquique y musgo, todo previamente esterilizado y cerrado con plástico y alambre de cobre. El ciclo fue isospórico con desarrollo del protalo tipoAdiantum. En la fase cordada se apreciaron gametófitos con anteridios y arquegonio en una proporción cercana al 70% y el otro 30% fueron gametófitos apogámicos. Los esporófitos jóvenes se obtuvieron a partir de los 63 días.

  11. Secondary metabolites with ecologic and medicinal implications in Anthemis cretica subsp. petraea from Majella National Park

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    Alessandro Venditti

    2016-11-01

    Full Text Available Anthemis cretica subsp. petraea (Ten. Greuter is a plant belonging to the Asteraceae family and endemic of central Italy. In this paper, the first analysis of the ethanolic fraction of samples collected in the Majella National Park is reported. Seven compounds were isolated and identified namely parthenolide (1, 9α-acetoxyparthenolide (2, tamarixetin (3, 7-hydroxycoumarin (4, 4'-hydroxyacetophenone (5, leucanthemitol (conduritol F (6, and proto-quercitol (7. Isolation of the compounds was achieved by means of column chromatography (CC, while their identification was achieved through spectroscopic and spectrometric techniques. The presence of these compounds is of great relevance. Compounds 1 and 2 are chemosystematic markers of the family, thus confirming the correct botanical classification of the species. Conversely, compounds 3, 5, and 7 were identified for the first time in the species and, instead, confirm the tendency of endemic entities to develop characteristic metabolite patterns in respect to cosmopolite species. Moreover, the presence of compounds 6 and 7 has ecologic implications and may be linked to this taxon’s adaption to dry environments. The production of these osmolytes may, in fact, represent the reason why this species is able to survive in extreme conditions of aridity. Lastly, from a medicinal standpoint, the isolated compounds are endowed with interesting biological activities and may justify, on a molecular base, the widespread traditional uses of the Anthemis species, as well as a basis for the use of the subspecies petraea.

  12. Expression of chitinase-encoding genes in Bacillus thuringiensis and toxicity of engineered B. thuringiensis subsp. aizawai toward Lymantria dispar larvae.

    Science.gov (United States)

    Lertcanawanichakul, Monthon; Wiwat, Chanpen; Bhumiratana, Amaret; Dean, Donald Harry

    2004-03-01

    Chitinase genes from Aeromonas hydrophila and Bacillus circulans No.4.1 were cloned into the plasmid pHY300PLK and designated as pHYA2 and pHYB43, respectively. Both plasmids were introduced into various strains of B. thuringiensis by electroporation. Plasmid pHYB43 was generally structurally stable, but showed lower segregrational stability than pHYA2 in B. thuringiensis subsp. aizawai when grown under nonselective conditions. The production of chitinase from B. thuringiensis subsp. aizawai harboring pHYB43 or pHYA2 could be detected after native polyacrylamide gel electrophoresis by using 4-methylumbelliferyl beta-D- N,N'- diacetylchitobioside as the substrate. Moreover, B. thuringiensis subsp. aizawai harboring pHYB43 gave 15 times higher chitinase activity than when harboring pHYA2, as determined by means of a colorimetric method using glycol chitin as the substrate. In addition, B. thuringiensis subsp. aizawai harboring pHYB43 was more toxic to gypsy moth larvae ( Lymantria dispar) than parental B. thuringiensis subsp. aizawai or its clone harboring pHYA2.

  13. Estudo dos parâmetros da ultrafiltração de permeado de soro de queijo fermentado por Lactococcus lactis subsp. lactis Ultrafiltration conditions of whey permeate fermented by Lactococcus lactis subsp. lactis

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    Viviane BRONSTEIN

    1998-04-01

    Full Text Available Permeado de soro doce, suplementado com extrato de levedura e peptona, foi utilizado como meio de crescimento para Lactococcus lactis subsp. lactis. No final da fase exponencial de crescimento, o meio de cultura fermentado foi submetido a uma ultrafiltração com o objetivo de concentrar o microrganismo. Foram realizados 6 processamentos diferentes, nos quais variou-se as condições iniciais da ultrafiltração, tendo sido avaliados os seguintes parâmetros: porosidade da membrana, pH e número de células viáveis no permeado e no retentado, a fim de ser estudado a influência de cada parâmetro na taxa de permeação da ultrafiltração. As membranas utilizadas foram eficazes como meio de barragem para o microrganismo Lactococcus lactis subsp. lactis, ficando o retentado com uma média celular de 10(8 ufc/ml e o permeado com uma média celular de 10² ufc/ml. Membranas de diferentes porosidades tiveram taxas de fluxo semelhantes. O aumento da concentração celular provocou a diminuição do fluxo. O pH também influenciou a taxa de permeação, havendo um aumento do fluxo quando foi utilizado um pH inicial mais alto.Cheese whey permeate supplemented with yeast extract and peptone was used as a growth medium for the bacteria Lactococcus lactis subsp. lactis. At the end of the exponential growth phase, the fermented growth medium was ultrafiltered to concentrate the microorganism and to evaluate the effect of the membrane porosity, inicial UF pH and cellular concentration in permeation rate during the ultrafiltration process. The membranes used were efficient as a mean of a barrage for the Lactococcus lactis subsp. lactis. On average, the cellular concentrations were 10(8 CFU/mL and 10² CFU/mL for retentate and permeate, respectively. Membranes of different porosities had very similar flux rates. Better flow rates were obtained with inicial UF pH 6,5 and with the minors micrrorganism concentration.

  14. Biosystematic studies on Dactylis L. 2. Personal research. 2.1. Morphological differentiation and occurrence of representatives of the genus Dactylis in Poland. 2.1.3. Distribution of D. glomerata subsp. aschersoniana (Graebn. Thell. in Poland

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    Marta Mizianty

    2014-01-01

    Full Text Available In this paper the author presents the distribution of D. g. subsp. aschersoniana (Graebn. Thell. in 198 localities in Poland. The studies on distribution were based mainly on revision and verification of materials from Polish herbaria. It is stated that subsp. aschersoniana occurs in deciduous forests in lowlands and on plateaus as well as in lower mountain altitudes. Part of the localities are situated beyond the present eastern limit of the Fagus sylvatica distribution. This confirms the opinion that this subspecies is not connected with beech woods but rather with oak-hornbeam forests. Sometimes subsp. glomerata, subsp. slovenica and subsp. aschersoniana were found in one locality. This fact is very important in the context of gene-flow between these three subspecies.

  15. [The relationship of Campylobacter jejuni subsp. jejuni enterotoxigenicity and the increase of cAMP and electrolyte changes in the rat intestine].

    Science.gov (United States)

    Fernández, H; Toro, J

    1998-08-01

    Small intestine alterations produced by the enterotoxigenic capacity of Campylobacter jejuni subsp. jejuni are similar to the hydric, electrolytic and pathological changes caused by choleraic and thermolabile Escherichia coli toxins. To study the enterotoxigenic capacity of 4 strains of Campylobacter jejuni subsp. jejuni using the intestinal loop model. Rat intestinal loops were inoculated with culture filtrates of the four strains. Enterotoxigenicity was assessed by fluid accumulation, the increase in Na+ and Cl- in the loop fluid, and cAMP increase in loop tissues. An enterotoxigenic Escherichia coil strain and sterile Brucella both were used as positive and negative controls, respectively. The filtrates of two strains produced fluid accumulation in the loops, significantly increased Na+ and Cl- secretion to the intestinal lumen and increased tissue cAMP levels. Some strains of Campylobacter jejuni subsp. jejuni are able to show enterotoxigenicity in vivo, increasing cAMP levels in the intestinal cells and altering electrolyte exchange mechanisms.

  16. The chromosome map of Bacillus thuringiensis subsp. canadensis HD224 is highly similar to that of the Bacillus cereus type strain ATCC 14579.

    Science.gov (United States)

    Carlson, C R; Johansen, T; Kolstø, A B

    1996-08-01

    A physical map of the Bacillus thuringiensis subsp. canadensis HD224 chromosome based on AscI, NotI, and SfiI restriction sites has been established. The chromosome map of 4.3 Mb was similar to a revised map of the chromosome of the B. cereus type strain ATCC 14579, except that the B. thuringiensis subsp. canadensis HD224 chromosome lacked a NotI site and had two additional AscI sites. The positions of 27 probes were identical in the common macromap. A probe for the insecticidal toxin gene, cryIA, hybridized only to the B. thuringiensis subsp. canadensis HD224 chromosome. The BssHII ribotype patterns were almost identical confirming the similarity between the two strains.

  17. Antiphagocytic function of an IgG glycosyl hydrolase from Streptococcus equi subsp. equi and its use as a vaccine component.

    Science.gov (United States)

    Flock, Margareta; Frykberg, Lars; Sköld, Markus; Guss, Bengt; Flock, Jan-Ingmar

    2012-08-01

    EndoSe from Streptococcus equi subsp. equi is an enzyme hydrolyzing glycosyl groups on IgG, analogous to EndoS from Streptococcus pyogenes. We here show that the activity of EndoSe leads to an antiphagocytic function and may thus be a contributory factor to immune evasion of S. equi. Despite the damaging effect that EndoSe has on IgG, antibodies against EndoSe can neutralize its function. Antibodies against EndoSe restored the opsonic activity of specific opsonizing antibodies. Mice infected with either S. equi subsp. equi or subsp. zooepidemicus or S. pyogenes could be protected by vaccination with EndoSe. It is speculated that EndoSe could be a suitable vaccine candidate against streptococcal infections.

  18. Essential Oil Composition of Three Centaurea Species from Turkey: Centaurea aggregata Fisch. & Mey. ex. DC. subsp. aggregata, C. balsamita Lam. and C. behen L.

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    Tuğçe Erdoğan

    2017-01-01

    Full Text Available The essential oils of Centaurea aggregata subsp. aggregata, C. balsamita and C. behen collected in the same habitat from Turkey, have been investigated. A total of 26, 54 and 26 components were identified in C. aggregata subsp. aggregata, C. balsamita and C. behen, respectively. The main constituents of the investigated populations of the three taxa have been revealed as follows: Centaurea aggregata subsp. aggregata: hexadecanoic acid (35.8%, phytol (7.2%, caryophyllene oxide(6.0%, spathulenol (6.0%; C. balsamita: hexadecanoic acid (23.0%, spathulenol (8.9%, germacrene D (2.1%; C. behen: hexadecanoic acid (32.7%, germacrene D (14.8%, and phytol (12.3%.

  19. Tingkat Keterjadian Penyakit Layu Stewart pada Benih dan Respon beberapa Varietas Jagung terhadap Infeksi Pantoea stewartii subsp. stewartii

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    Haliatur Rahma

    2014-03-01

    Full Text Available Disease incidence of Stewart’s wilt on the seed and response of several maize varieties to Pantoea stewartii subp. stewartii. Stewart’s wilt disease of maize is caused by Pantoea stewartii subsp. stewartii.  This bacterium is seed-borne pathogens, when attacked maize caused yield lost 40-100%.  The objective of this research was to detemine the incidence level of stewart’s wilt disease, growth of some varieties of maize and their response to stewart’s wilt pathogens Pantoea stewartii subsp. stewartii.  The research was conducted in the Laboratory of Bacteriology and Greenhouse Cikabayan IPB from November 2011 to March 2012. In experiment I, nineteen samples of maize were used for symptom test  in the maize seedling stage, using Randomized Block Design with three replications. Experiment II used a Randomized Block Design with 2 factors: maize varieties (8 hybrids varieties, 3 open pollinated varieties, and 7 sweet corn varieties and bacteria isolates  (BGR 2, BGR 4, BGR 28, BGR7 and PSM 27, with three replications. The results showed in experiment I, the incidence of stewart’s wilt disease ranged 2.00 – 15.33%, germination and vigor index of maize seed were 68.00 – 95.33% and 55.33 – 90.67% respectively.  While in experiment II, hybrid and open pollinated of maize varieties were resistant to moderately susceptible   while all sweet corn varieties were susceptible to infection of Pantoea stewartii subsp. stewartii.

  20. Differential expression of in vivo and in vitro protein profile of outer membrane of Acidovorax avenae subsp. avenae.

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    Muhammad Ibrahim

    Full Text Available Outer membrane (OM proteins play a significant role in bacterial pathogenesis. In this work, we examined and compared the expression of the OM proteins of the rice pathogen Acidovorax avenae subsp. avenae strain RS-1, a Gram-negative bacterium, both in an in vitro culture medium and in vivo rice plants. Global proteomic profiling of A. avenae subsp. avenae strain RS-1 comparing in vivo and in vitro conditions revealed the differential expression of proteins affecting the survival and pathogenicity of the rice pathogen in host plants. The shotgun proteomics analysis of OM proteins resulted in the identification of 97 proteins in vitro and 62 proteins in vivo by mass spectrometry. Among these OM proteins, there is a high number of porins, TonB-dependent receptors, lipoproteins of the NodT family, ABC transporters, flagellins, and proteins of unknown function expressed under both conditions. However, the major proteins such as phospholipase and OmpA domain containing proteins were expressed in vitro, while the proteins such as the surface anchored protein F, ATP-dependent Clp protease, OmpA and MotB domain containing proteins were expressed in vivo. This may indicate that these in vivo OM proteins have roles in the pathogenicity of A. avenae subsp. avenae strain RS-1. In addition, the LC-MS/MS identification of OmpA and MotB validated the in silico prediction of the existance of Type VI secretion system core components. To the best of our knowledge, this is the first study to reveal the in vitro and in vivo protein profiles, in combination with LC-MS/MS mass spectra, in silico OM proteome and in silico genome wide analysis, of pathogenicity or plant host required proteins of a plant pathogenic bacterium.

  1. Differential expression of in vivo and in vitro protein profile of outer membrane of Acidovorax avenae subsp. avenae.

    Science.gov (United States)

    Ibrahim, Muhammad; Shi, Yu; Qiu, Hui; Li, Bin; Jabeen, Amara; Li, Liping; Liu, He; Kube, Michael; Xie, Guanlin; Wang, Yanli; Blondel, Carlos; Santiviago, Carlos A; Contreras, Ines; Sun, Guochang

    2012-01-01

    Outer membrane (OM) proteins play a significant role in bacterial pathogenesis. In this work, we examined and compared the expression of the OM proteins of the rice pathogen Acidovorax avenae subsp. avenae strain RS-1, a Gram-negative bacterium, both in an in vitro culture medium and in vivo rice plants. Global proteomic profiling of A. avenae subsp. avenae strain RS-1 comparing in vivo and in vitro conditions revealed the differential expression of proteins affecting the survival and pathogenicity of the rice pathogen in host plants. The shotgun proteomics analysis of OM proteins resulted in the identification of 97 proteins in vitro and 62 proteins in vivo by mass spectrometry. Among these OM proteins, there is a high number of porins, TonB-dependent receptors, lipoproteins of the NodT family, ABC transporters, flagellins, and proteins of unknown function expressed under both conditions. However, the major proteins such as phospholipase and OmpA domain containing proteins were expressed in vitro, while the proteins such as the surface anchored protein F, ATP-dependent Clp protease, OmpA and MotB domain containing proteins were expressed in vivo. This may indicate that these in vivo OM proteins have roles in the pathogenicity of A. avenae subsp. avenae strain RS-1. In addition, the LC-MS/MS identification of OmpA and MotB validated the in silico prediction of the existance of Type VI secretion system core components. To the best of our knowledge, this is the first study to reveal the in vitro and in vivo protein profiles, in combination with LC-MS/MS mass spectra, in silico OM proteome and in silico genome wide analysis, of pathogenicity or plant host required proteins of a plant pathogenic bacterium.

  2. Bioactive Compounds of Endemic Species Sideritis raeseri subsp. raeseri Grown in National Park Galičica

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    Nebojša Menković

    2013-05-01

    Full Text Available Collection of Sideritis raeseri subsp. raeseri has a long tradition in local communities in the Ohrid-Prespa region. The aim of the present study was the analysis of bioactive compounds especially those with anti-inflammatory activity. Combination of the UV and MS data allowed the characterization of 17 compounds, which could be classified into flavonoid glycosides or hydroxycynnamic acid derivatives. Six of them were isolated using preparative HPLC: isoscutellarein 7-O-[6′″-O-acetyl- β -D-allopyranosyl-(1→2]- β -D-glucopyranoside, 4′-O-methylhypolaetin 7-O-[6′″-O-acetyl- β -D-allopyranosyl-(1→2]- β -D-glucopyranoside, hypolaetin 7-O-[6′″-O-acetyl- β -D-allopyranosyl-(1→2]-6″-O-acetyl- β -D-glucopyranoside, 4′-O-methylisoscutellarein 7-O-[6′″-O-acetyl- β -D-allopyranosyl-(1→2]- β -D-glucopyranoside, isoscutellarein 7-O-[6′″-O-acetyl- β -D-allopyranosyl-(1→2]-6″-O-acetyl- β -D-glucopyranoside and 4′-O-methylhypolaetin 7-O-[6′″-O-acetyl- β -D-allopyranosyl-(1→2]-6″-O-acetyl- β -D-glucopyranoside. The presence of phenylpropanoid glycoside martynoside was reported for the first time in the wild growing S. raeseri subsp. raeseri. Hypolaetin derivatives, known for their antiinflammatory activity, dominated and were more abundant in S. raeseri subsp. raeseri grown in NPG in comparison with S. scardica grown nearby the NPG. The type of solvent affected distribution and the amount of bioactive compounds and the advantage was given to less polar extracts which were richer in hypolaetin derivatives.

  3. The mosaic architecture of Aeromonas salmonicida subsp. salmonicida pAsa4 plasmid and its consequences on antibiotic resistance

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    Katherine H. Tanaka

    2016-10-01

    Full Text Available Aeromonas salmonicida subsp. salmonicida, the causative agent of furunculosis in salmonids, is an issue especially because many isolates of this bacterium display antibiotic resistances, which limit treatments against the disease. Recent results suggested the possible existence of alternative forms of pAsa4, a large plasmid found in A. salmonicida subsp. salmonicida and bearing multiple antibiotic resistance genes. The present study reveals the existence of two newly detected pAsa4 variants, pAsa4b and pAsa4c. We present the extensive characterization of the genomic architecture, the mobile genetic elements and the antimicrobial resistance genes of these plasmids in addition to the reference pAsa4 from the strain A449. The analysis showed differences between the three architectures with consequences on the content of resistance genes. The genomic plasticity of the three pAsa4 variants could be partially explained by the action of mobile genetic elements like insertion sequences. Eight additional isolates from Canada and Europe that bore similar antibiotic resistance patterns as pAsa4-bearing strains were genotyped and specific pAsa4 variants could be attributed to phenotypic profiles. pAsa4 and pAsa4c were found in Europe, while pAsa4b was found in Canada. In accordance with their content in conjugative transfer genes, only pAsa4b and pAsa4c can be transferred by conjugation in Escherichia coli. The plasticity of pAsa4 variants related to the acquisition of antibiotic resistance indicates that these plasmids may pose a threat in terms of the dissemination of antimicrobial-resistant A. salmonicida subsp. salmonicida bacteria.

  4. Screening in a Lactobacillus delbrueckii subsp. bulgaricus collection to select a strain able to survive to the human intestinal tract.

    Science.gov (United States)

    Vázquez, Clotilde; Botella-Carretero, José I; García-Albiach, Raimundo; Pozuelo, María J; Rodríguez-Baños, Mercedes; Baquero, Fernando; Baltadjieva, María A; del Campo, Rosa

    2013-01-01

    Genetic diversity and resistance of Lactobacillus bulgaricus sbsp. delbrueckii collection with 100 isolates from different home-made yogurt in rural Bulgarian areas were determined. The strain K98 was the most resistant to bile salts and low pH. Survival and effects on short chain fatty acids production were tested in 20 healthy volunteers. High genetic diversity was observed in the L. bulgaricus collection by RAPD, whereas the ability of tolerate high deoxycholic acid concentrations, and different acid pHs was variable. The strain K98 was selected and used to prepare a homemade yogurt which was administered to 20 healthy volunteers (500 ml/day during 15d). A basal faecal sample and another after yogurt intake were recovered. DGGE experiments, using both universal and Lactic Acid Bacteria (LAB) primers, demonstrated no significant changes in the qualitative composition of gut microbiota. A band corresponding to L. bulgaricus was observed in all 20 samples. Viable L. bulgaricus K98 strain was only recovered in one volunteer. After yogurt intake we found an increase of LAB and Clostridium perfringens, and a decrease of Bacteroides- Prevotella-Porphyromonas. In addition, increases of acetic, butyric and 2-hydroxy-butyric acids in faeces were detected. Genetic diversity of L. delbrueckii subsp. bulgaricus especie is high We have isolated a probiotic resistant strain to bile and high acidity, L. delbrueckii subsp. bulgaricus-K98. Qualitative and quantitative changes in the intestinal microbiota are found after ingestion of a homemade yogurt containing this strain, with a concomitant increase in faecal SCFA. Our findings support the interest in developing further studies providing different amounts of L. delbrueckii subsp. bulgaricus-K98, and should evaluate its clinical effects in human disease. Copyright © AULA MEDICA EDICIONES 2013. Published by AULA MEDICA. All rights reserved.

  5. Bifidobacterium animalis subsp lactis CNCM-I2494 restores gut barrier permeability in chronically low-grade inflamed mice

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    Rebeca eMartín

    2016-05-01

    Full Text Available Growing evidence supports the efficacy of many probiotic strains in the management of gastrointestinal disorders associated with deregulated intestinal barrier function and/or structure. In particular, bifidobacteria have been studied for their efficacy to both prevent and treat a broad spectrum of animal and/or human gut disorders. The aim of the current work was thus to evaluate effects on intestinal barrier function of Bifidobacterium animalis subsp lactis CNCM-I2494, a strain used in fermented dairy products. A chronic DNBS-induced low-grade inflammation model causing gut dysfunction in mice was used in order to study markers of inflammation, intestinal permeability and immune function in the presence of the bacterial strain. In this chronic low-grade inflammation mice model several parameters pointed out the absence of an over active inflammation process. However, gut permeability, lymphocyte populations and colonic cytokines were found to be altered. B. animalis subsp lactis CNCM-I2494 was able to protect barrier functions by restoring intestinal permeability, colonic goblet cell populations and cytokine levels. Furthermore, tight junction (TJ proteins levels were also measured by qRT-PCR showing the ability of this strain to specifically normalize the level of several TJ proteins, in particular for claudin-4. Finally, B. lactis strain counterbalanced CD4+ lymphocyte alterations in both spleen and mesenteric lymphoid nodes (MLN. It restores the Th1/Th2 ratio altered by the DNBS challenge (which locally augments CD4+ Th1 cells by increasing the Th2 response as measured by the increase in the production of major representative Th2 cytokines (IL-4, IL-5 and IL-10. Altogether, these data suggest that B. animalis subsp lactis CNCM-I2494 may efficiently prevent disorders associated with increased barrier permeability.

  6. Molecular characterization of a DNA fragment harboring the replicon of pBMB165 from Bacillus thuringiensis subsp. tenebrionis

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    Yu Ziniu

    2006-10-01

    Full Text Available Abstract Background Bacillus thuringiensis belongs to the Bacillus cereus sensu lato group of Gram-positive and spore-forming bacteria. Most isolates of B. thuringiensis can bear many endogenous plasmids, and the number and size of these plasmids can vary widely among strains or subspecies. As far as we know, the replicon of the plasmid pBMB165 is the first instance of a plasmid replicon being isolated from subsp. tenebrionis and characterized. Results A 20 kb DNA fragment containing a plasmid replicon was isolated from B. thuringiensis subsp. tenebrionis YBT-1765 and characterized. By Southern blot analysis, this replicon region was determined to be located on pBMB165, the largest detected plasmid (about 82 kb of strain YBT-1765. Deletion analysis revealed that a replication initiation protein (Rep165, an origin of replication (ori165 and an iteron region were required for replication. In addition, two overlapping ORFs (orf6 and orf10 were found to be involved in stability control of plasmid. Sequence comparison showed that the replicon of pBMB165 was homologous to the pAMβ1 family replicons, indicating that the pBMB165 replicon belongs to this family. The presence of five transposable elements or remnants thereof in close proximity to and within the replicon control region led us to speculate that genetic exchange and recombination are potentially responsible for the divergence among the replicons of this plasmid family. Conclusion The replication and stability features of the pBMB165 from B. thuringiensis subsp. tenebrionis YBT-1765 were identified. Of particular interest is the homology and divergence shared between the pBMB165 replicon and other pAMβ1 family replicons.

  7. Interaction of Francisella noatunensis subsp. orientalis with Oreochromis mossambicus bulbus arteriosus cell line

    Science.gov (United States)

    Soto, Esteban; Yun, Susan; Lewis, J.; Kearney, Michael T.; Hansen, John D.

    2017-01-01

    Francisella noatunensis subsp. orientalis (Fno) (syn. F. asiatica) is an emergent warmwater fish pathogen and the causative agent of piscine francisellosis. Although Fno causes septicemia and can live extracellularly in infected tilapia (Oreochromis spp.), the early interaction of Fno with vasculature endothelium is unknown. In the present study, we examined the interaction of wild-type Fno (WT) and two Fno knockout [intracellular growth loci C (ΔiglC) and pathogenicity determinant protein A (ΔpdpA)] strains with the endothelial O. mossambicus bulbus arteriosus cell line (TmB) at 25 °C and 30 °C. Similar amounts of WT, ΔiglC, and ΔpdpA attached and were detected intracellularly after 5 h of incubation at both temperatures; however temperature affected attachment and uptake. While significantly greater amounts of Fno (WT, ΔiglC, and ΔpdpA) were detected intracellularly when TmB cells were incubated at 30 °C, bacteria attached to TmBs at greater levels at 25 °C. Only WT Fno was able to replicate intracellularly at 25 °C, which resulted in Fno mediated cytotoxicity and apoptosis at 24 and 72 h post-infection. WT Fno incubated at 30 °C as well as ΔiglC,and ΔpdpA incubated at 25 °C and 30 °C were all defective for survival, replication, and the ability to cause cytotoxicity in TmB. Taken together, these results demonstrate that temperature plays a vital role for Fno intracellular survival, persistence and cytotoxicity.

  8. Genome-Wide Association Mapping of Stem Rust Resistance in Hordeum vulgare subsp. spontaneum

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    Ahmad H. Sallam

    2017-10-01

    Full Text Available Stem rust was one of the most devastating diseases of barley in North America. Through the deployment of cultivars with the resistance gene Rpg1, losses to stem rust have been minimal over the past 70 yr. However, there exist both domestic (QCCJB and foreign (TTKSK aka isolate Ug99 pathotypes with virulence for this important gene. To identify new sources of stem rust resistance for barley, we evaluated the Wild Barley Diversity Collection (WBDC (314 ecogeographically diverse accessions of Hordeum vulgare subsp. spontaneum for seedling resistance to four pathotypes (TTKSK, QCCJB, MCCFC, and HKHJC of the wheat stem rust pathogen (Puccinia graminis f. sp. tritici, Pgt and one isolate (92-MN-90 of the rye stem rust pathogen (P. graminis f. sp. secalis, Pgs. Based on a coefficient of infection, the frequency of resistance in the WBDC was low ranging from 0.6% with HKHJC to 19.4% with 92-MN-90. None of the accessions was resistant to all five cultures of P. graminis. A genome-wide association study (GWAS was conducted to map stem rust resistance loci using 50,842 single-nucleotide polymorphic markers generated by genotype-by-sequencing and ordered using the new barley reference genome assembly. After proper accounting for genetic relatedness and structure among accessions, 45 quantitative trait loci were identified for resistance to P. graminis across all seven barley chromosomes. Three novel loci associated with resistance to TTKSK, QCCJB, MCCFC, and 92-MN-90 were identified on chromosomes 5H and 7H, and two novel loci associated with resistance to HKHJC were identified on chromosomes 1H and 3H. These novel alleles will enhance the diversity of resistance available for cultivated barley.

  9. Discovery of novel small molecule modulators of Clavibacter michiganensis subsp. michiganensis

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    Xiulan eXu

    2015-10-01

    Full Text Available Clavibacter michiganensis subsp. michiganensis (Cmm is a Gram-positive seed-transmitted bacterial phytopathogen responsible for substantial economic losses by adversely affecting tomato production worldwide. A high-throughput, cell-based screen was adapted to identify novel small molecule growth inhibitors to serve as leads for future bactericide development. A library of 4,182 compounds known to be bioactive against Saccharomyces cerevisiae was selected for primary screening against Cmm wild-type strain C290 for whole-cell growth inhibition. Four hundred sixty-eight molecules (11.2% hit rate were identified as bacteriocidal or bacteriostatic against Cmm at 200 M. Seventy-seven candidates were selected based on Golden Triangle analyses for secondary screening. Secondary screens showed that several of these candidates were strain-selective. Several compounds were inhibitory to multiple Cmm strains as well as Bacillus subtilis, but not Pseudomonas fluorescens, Mitsuaria sp., Lysobacter enzymogenes, Lactobacillus rhamnosus, Bifidobacter animalis, or Escherichia coli. Most of the compounds were not phytotoxic and did not show overt host toxicity. Using a novel 96-well bioluminescent Cmm seedling infection assay, we assessed effects of selected compounds on pathogen infection. The 12 most potent novel molecules were identified by compiling the scores from all secondary screens combined with the reduction of pathogen infection in planta. When tested for ability to develop resistance to the top-12 compounds, no resistant Cmm were recovered, suggesting that the discovered compounds are unlikely to induce resistance. In conclusion, here we report top-12 compounds that provide chemical scaffolds for future Cmm-specific bactericide development.

  10. Discovery of novel small molecule modulators of Clavibacter michiganensis subsp. michiganensis.

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    Xu, Xiulan; Kumar, Anand; Deblais, Loïc; Pina-Mimbela, Ruby; Nislow, Corey; Fuchs, James R; Miller, Sally A; Rajashekara, Gireesh

    2015-01-01

    Clavibacter michiganensis subsp. michiganensis (Cmm) is a Gram-positive seed-transmitted bacterial phytopathogen responsible for substantial economic losses by adversely affecting tomato production worldwide. A high-throughput, cell-based screen was adapted to identify novel small molecule growth inhibitors to serve as leads for future bactericide development. A library of 4,182 compounds known to be bioactive against Saccharomyces cerevisiae was selected for primary screening against Cmm wild-type strain C290 for whole-cell growth inhibition. Four hundred sixty-eight molecules (11.2% hit rate) were identified as bacteriocidal or bacteriostatic against Cmm at 200 μM. Seventy-seven candidates were selected based on Golden Triangle analyses for secondary screening. Secondary screens showed that several of these candidates were strain-selective. Several compounds were inhibitory to multiple Cmm strains as well as Bacillus subtilis, but not to Pseudomonas fluorescens, Mitsuaria sp., Lysobacter enzymogenes, Lactobacillus rhamnosus, Bifidobacterium animalis, or Escherichia coli. Most of the compounds were not phytotoxic and did not show overt host toxicity. Using a novel 96-well bioluminescent Cmm seedling infection assay, we assessed effects of selected compounds on pathogen infection. The 12 most potent novel molecules were identified by compiling the scores from all secondary screens combined with the reduction of pathogen infection in planta. When tested for ability to develop resistance to the top-12 compounds, no resistant Cmm were recovered, suggesting that the discovered compounds are unlikely to induce resistance. In conclusion, here we report top-12 compounds that provide chemical scaffolds for future Cmm-specific bactericide development.

  11. Metabolomic profiling in cattle experimentally infected with Mycobacterium avium subsp. paratuberculosis.

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    Jeroen De Buck

    Full Text Available The sensitivity of current diagnostics for Johne's disease, a slow, progressing enteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP, is too low to reliably detect all infected animals in the subclinical stage. The objective was to identify individual metabolites or metabolite profiles that could be used as biomarkers of early MAP infection in ruminants. In a monthly follow-up for 17 months, calves infected at 2 weeks of age were compared with aged-matched controls. Sera from all animals were analyzed by 1H nuclear magnetic resonance spectrometry. Spectra were acquired, processed, and quantified for analysis. The concentration of many metabolites changed over time in all calves, but some metabolites only changed over time in either infected or non-infected groups and the change in others was impacted by the infection. Hierarchical multivariate statistical analysis achieved best separation between groups between 300 and 400 days after infection. Therefore, a cross-sectional comparison between 1-year-old calves experimentally infected at various ages with either a high- or a low-dose and age-matched non-infected controls was performed. Orthogonal Projection to Latent Structures Discriminant Analysis (OPLS DA yielded distinct separation of non-infected from infected cattle, regardless of dose and time (3, 6, 9 or 12 months after infection. Receiver Operating Curves demonstrated that constructed models were high quality. Increased isobutyrate in the infected cattle was the most important agreement between the longitudinal and cross-sectional analysis. In general, high- and low-dose cattle responded similarly to infection. Differences in acetone, citrate, glycerol and iso-butyrate concentrations indicated energy shortages and increased fat metabolism in infected cattle, whereas changes in urea and several amino acids (AA, including the branched chain AA, indicated increased protein turnover. In conclusion, metabolomics

  12. The Science behind the Probiotic Strain Bifidobacterium animalis subsp. lactis BB-12®

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    Jungersen, Mikkel; Wind, Anette; Johansen, Eric; Christensen, Jeffrey E.; Stuer-Lauridsen, Birgitte; Eskesen, Dorte

    2014-01-01

    This review presents selected data on the probiotic strain Bifidobacterium animalis subsp. lactis BB-12® (BB-12®), which is the world’s most documented probiotic Bifidobacterium. It is described in more than 300 scientific publications out of which more than 130 are publications of human clinical studies. The complete genome sequence of BB-12® has been determined and published. BB-12® originates from Chr. Hansen’s collection of dairy cultures and has high stability in foods and as freeze dried powders. Strain characteristics and mechanisms of BB-12® have been established through extensive in vitro testing. BB-12® exhibits excellent gastric acid and bile tolerance; it contains bile salt hydrolase, and has strong mucus adherence properties, all valuable probiotic characteristics. Pathogen inhibition, barrier function enhancement, and immune interactions are mechanisms that all have been demonstrated for BB-12®. BB-12® has proven its beneficial health effect in numerous clinical studies within gastrointestinal health and immune function. Clinical studies have demonstrated survival of BB-12® through the gastrointestinal tract and BB-12® has been shown to support a healthy gastrointestinal microbiota. Furthermore, BB-12® has been shown to improve bowel function, to have a protective effect against diarrhea, and to reduce side effects of antibiotic treatment, such as antibiotic-associated diarrhea. In terms of immune function, clinical studies have shown that BB-12® increases the body’s resistance to common respiratory infections as well as reduces the incidence of acute respiratory tract infections. PMID:27682233

  13. Identification and Phytotoxicity Assessment of Phenolic Compounds in Chrysanthemoides monilifera subsp. monilifera (Boneseed.

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    Md Abdullah Yousuf Al Harun

    Full Text Available Chrysanthemoides monilifera subsp. monilifera (boneseed, a weed of national significance in Australia, threatens indigenous species and crop production through allelopathy. We aimed to identify phenolic compounds produced by boneseed and to assess their phytotoxicity on native species. Phenolic compounds in water and methanol extracts, and in decomposed litter-mediated soil leachate were identified using HPLC, and phytotoxicity of identified phenolics was assessed (repeatedly through a standard germination bioassay on native Isotoma axillaris. The impact of boneseed litter on native Xerochrysum bracteatum was evaluated using field soil in a greenhouse. Collectively, we found the highest quantity of phenolic compounds in boneseed litter followed by leaf, root and stem. Quantity varied with extraction media. The rank of phenolics concentration in boneseed was in the order of ferulic acid > phloridzin > catechin > p-coumaric acid and they inhibited germination of I. axillaris with the rank of ferulic acid > catechin > phloridzin > p-coumaric acid. Synergistic effects were more severe compared to individual phenolics. The litter-mediated soil leachate (collected after15 days exhibited strong phytotoxicity to I. axillaris despite the level of phenolic compounds in the decomposed leachate being decreased significantly compared with their initial level. This suggests the presence of other unidentified allelochemicals that individually or synergistically contributed to the phytotoxicity. Further, the dose response phytotoxic impacts exhibited by the boneseed litter-mediated soil to native X. bracteatum in a more naturalistic greenhouse experiment might ensure the potential allelopathy of other chemical compounds in the boneseed invasion. The reduction of leaf relative water content and chlorophyll level in X. bracteatum suggest possible mechanisms underpinning plant growth inhibition caused by boneseed litter allelopathy. The presence of a substantial

  14. Strand-specific RNA-seq analysis of the Lactobacillus delbrueckii subsp. bulgaricus transcriptome.

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    Zheng, Huajun; Liu, Enuo; Shi, Tao; Ye, Luyi; Konno, Tomonobu; Oda, Munehiro; Ji, Zai-Si

    2016-02-01

    Lactobacillus delbrueckii subsp. bulgaricus 2038 (Lb. bulgaricus 2038) is an industrial bacterium that is used as a starter for dairy products. We proposed several hypotheses concerning its industrial features previously. Here, we utilized RNA-seq to explore the transcriptome of Lb. bulgaricus 2038 from four different growth phases under whey conditions. The most abundantly expressed genes in the four stages were mainly involved in translation (for the logarithmic stage), glycolysis (for control/lag stages), lactic acid production (all the four stages), and 10-formyl tetrahydrofolate production (for the stationary stage). The high expression of genes like d-lactate dehydrogenase was thought as a result of energy production, and consistent expression of EPS synthesis genes, the restriction-modification (RM) system and the CRISPR/Cas system were validated for explaining the advantage of this strain in yoghurt production. Several postulations, like NADPH production through GapN bypass, converting aspartate into carbon-skeleton intermediates, and formate production through degrading GTP, were proved not working under these culture conditions. The high expression of helicase genes and co-expressed amino acids/oligopeptides transporting proteins indicated that the helicase might mediate the strain obtaining nitrogen source from the environment. The transport system of Lb. bulgaricus 2038 was found to be regulated by antisense RNA, hinting the potential application of non-coding RNA in regulating lactic acid bacteria (LAB) gene expression. Our study has primarily uncovered Lb. bulgaricus 2038 transcriptome, which could gain a better understanding of the regulation system in Lb. bulgaricus and promote its industrial application.

  15. Fabrication of a Novel Conductometric Biosensor for Detecting Mycobacterium avium subsp. paratuberculosis Antibodies

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    Steven Bolin

    2008-09-01

    Full Text Available Johne’s disease (JD is one of the most costly bacterial diseases in cattle. In the U.S., economic losses from the disease have been estimated to exceed $1,500,000,000 per year, mainly from the effects of reduced milk production. Current diagnostic tests for JD are laboratory based and many of those tests require specialized equipment and training. Development of rapid and inexpensive diagnostic assays, which are adapted for point-ofcare applications, would aid in the control of JD. In this study, a polyaniline (Pani-based conductometric biosensor, in an immunomigration format, was fabricated for the detection of serum antibody (IgG against the causal organism of JD, Mycobacterium avium subsp. paratuberculosis (MAP. Immobilized Mycobacterium avium purified proteins in the capture membrane were used to detect MAP IgG, previously bound with Pani/anti-bovine IgG* conjugate in the conjugate membrane. After detection, the Pani in the sandwiched captured complex bridges an electrical circuit between the silver electrodes, flanking the capture membrane. The electrical conductance, caused by Pani, was measured as drop in electrical resistance. Testing of the biosensor with known JD positive and negative serum samples demonstrated a significant difference in the mean resistance observed between the groups. This proof-of-concept study demonstrated that a conductometric biosensor could detect MAP IgG in 2 minutes. The biosensor’s speed of detection and the equipment involved would, among other things, support its application towards the various point-ofcare opportunities aimed at JD management and control.

  16. Dysbiosis of the Fecal Microbiota in Cattle Infected with Mycobacterium avium subsp. paratuberculosis.

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    Marie-Eve Fecteau

    Full Text Available Johne's disease (JD is a chronic, intestinal infection of cattle, caused by Mycobacterium avium subsp. paratuberculosis (MAP. It results in granulomatous inflammation of the intestinal lining, leading to malabsorption, diarrhea, and weight loss. Crohn's disease (CD, a chronic, inflammatory gastrointestinal disease of humans, has many clinical and pathologic similarities to JD. Dysbiosis of the enteric microbiota has been demonstrated in CD patients. It is speculated that this dysbiosis may contribute to the intestinal inflammation observed in those patients. The purpose of this study was to investigate the diversity patterns of fecal bacterial populations in cattle infected with MAP, compared to those of uninfected control cattle, using phylogenomic analysis. Fecal samples were selected to include samples from 20 MAP-positive cows; 25 MAP-negative herdmates; and 25 MAP-negative cows from a MAP-free herd. The genomic DNA was extracted; PCR amplified sequenced on a 454 Roche platform, and analyzed using QIIME. Approximately 199,077 reads were analyzed from 70 bacterial communities (average of 2,843 reads/sample. The composition of bacterial communities differed between the 3 treatment groups (P < 0.001; Permanova test. Taxonomic assignment of the operational taxonomic units (OTUs identified 17 bacterial phyla across all samples. Bacteroidetes and Firmicutes constituted more than 95% of the bacterial population in the negative and exposed groups. In the positive group, lineages of Actinobacteria and Proteobacteria increased and those of Bacteroidetes and Firmicutes decreased (P < 0.001. Actinobacteria was highly abundant (30% of the total bacteria in the positive group compared to exposed and negative groups (0.1-0.2%. Notably, the genus Arthrobacter was found to predominate Actinobacteria in the positive group. This study indicates that MAP-infected cattle have a different composition of their fecal microbiota than MAP-negative cattle.

  17. Different Mycobacterium avium subsp. paratuberculosis MIRU-VNTR patterns coexist within cattle herds.

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    van Hulzen, K J E; Heuven, H C M; Nielen, M; Hoeboer, J; Santema, W J; Koets, A P

    2011-03-24

    A better understanding of the biodiversity of Mycobacterium avium subsp. paratuberculosis (MAP) offers more insight in the epidemiology of paratuberculosis and therefore may contribute to the control of the disease. The aim of this study was to investigate the genetic diversity in bovine MAP isolates using PCR-based methods detecting genetic elements called Variable-Number Tandem Repeats (VNTRs) and Mycobacterial Interspersed Repetitive Units (MIRUs) to determine if multiple MAP strains can coexist on farms with endemic MAP infection. For 52 temporal isolates originating from infected cattle from 32 commercial dairy herds with known trading history, MIRU-VNTR analysis was applied at 10 loci of which six showed variation. Within the group of 52 isolates, 17 different MIRU-VNTR patterns were detected. One MIRU-VNTR pattern was found in 29 isolates, one pattern in four isolates, one pattern in three isolates, two times one MIRU-VNTR pattern was found occurring in two isolates, and 12 patterns were found only once. Eleven herds provided multiple isolates. In five herds a single MIRU-VNTR pattern was detected among multiple isolates whereas in six herds more than one pattern was found. This study confirms that between dairy farms as well as within dairy farms, infected animals shed MAP with different MIRU-VNTR patterns. Analysis of trading history and age within herds indicated that cows born within the same birth cohort can be infected with MAP strains exhibiting variations in the number of MIRU-VNTR repeats. These data indicate that such multiple genotypes of MAP can coexist within one herd. Copyright © 2010 Elsevier B.V. All rights reserved.

  18. Virulence of Mycobacterium avium Subsp. hominissuis Human Isolates in an in vitro Macrophage Infection Model

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    Laura Rindi

    2018-01-01

    Full Text Available Background: Mycobacterium avium subsp. hominissuis (MAH is an environmental opportunistic pathogen for humans and swine worldwide; in humans, the vast majority of MAH infections is due to strains belonging to specific genotypes, such as the internal transcribed spacer (ITS-sequevars Mav-A and Mav-B that mostly cause pulmonary infections in elderly patients and severe disseminated infections in acquired immunodeficiency syndrome patients, respectively. To test whether the different types of infections in distinct patients' populations might reflect a different virulence of the infecting genotypes, MAH human isolates, genotyped by ITS sequencing and MIRU-VNTR minisatellite analysis, were studied for the capacity to infect and replicate in human macrophages in vitro. Methods: Cultures of human peripheral blood mononuclear cells and phagocytic human leukemic cell line THP-1 cells were infected with each MAH isolate and intracellular colony-forming units (CFU were determined. Results: At 2 h after infection, i.e., immediately after cell entry, the numbers of intracellular bacteria did not differ between Mav-A and Mav-B organisms in both phagocytic cell types. At 5 days, Mav-A organisms, sharing highly related VNTR-MIRU genotypes, yielded numbers of intracellular CFUs significantly higher than Mav-B organisms in both phagocytic cell types. MIRU-VNTR-based minimum spanning tree analysis of the MAH isolates showed a divergent phylogenetic pathway of Mav-A and Mav-B organisms. Conclusion: Mav-A and Mav-B sequevars might have evolved different pathogenetic properties that might account for their association with different human infections.

  19. Prevalence of Francisella noatunensis subsp. orientalis in cultured tilapia on the island of Oahu, Hawaii.

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    Soto, Esteban; McGovern-Hopkins, Kathleen; Klinger-Bowen, Ruth; Fox, Bradley K; Brock, James; Antonio, Nathene; Waal, Zelda van der; Rushton, Stephen; Mill, Aileen; Tamaru, Clyde S

    2013-06-01

    Francisellosis is an emergent disease in cultured and wild aquatic animals. The causative agent, Francisella noatunensis subsp. orientalis (Fno), is a gram-negative bacterium recognized as one of the most virulent pathogens of warmwater fish. The main objective of this project was to investigate the prevalence of Fno in cultured tilapia (specifically, Mozambique Tilapia Oreochromis mossambicus, Koilapia [also known as Wami Tilapia] O. hornorum, Blue Tilapia O. aureus, and Nile Tilapia O. niloticus hybrids) on the island of Oahu, Hawaii, using conventional and real-time PCR assays followed by statistical modeling to compare the different diagnostic methods and identify potential risk factors. During 2010 and 2012, 827 fish were collected from different geographical locations throughout the island of Oahu. Upon collection of fish, the water temperature in the rearing system and the length of individual fish were measured. Extraction of DNA from different tissues collected aseptically during necropsy served as a template for molecular diagnosis. High correlation between both molecular methods was observed. Moreover, the bacterium was isolated from infected tilapia on selective media and confirmed to be Fno utilizing a species-specific Taqman-based real-time PCR assay. Although a direct comparison of the prevalence of Fno between the different geographical areas was not possible, the results indicate a high prevalence of Fno DNA in cultured tilapia throughout the farm sites located on Oahu. Of the different tilapia species and hybrids currently cultured in Hawaii, Mozambique Tilapia were more susceptible to infection than Koilapia. Water temperature in the rearing systems and fish size also had a strong effect on the predicted level of infection, with fish held at lower temperatures and smaller fish being more susceptible to piscine francisellosis.

  20. Development of a recombinase polymerase amplification assay for rapid detection of Francisella noatunensis subsp. orientalis.

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    Khalid Shahin

    Full Text Available Francisella noatunensis subsp. orientalis (Fno is the causative agent of piscine francisellosis in warm water fish including tilapia. The disease induces chronic granulomatous inflammation with high morbidity and can result in high mortality. Early and accurate detection of Fno is crucial to set appropriate outbreak control measures in tilapia farms. Laboratory detection of Fno mainly depends on bacterial culture and molecular techniques. Recombinase polymerase amplification (RPA is a novel isothermal technology that has been widely used for the molecular diagnosis of various infectious diseases. In this study, a recombinase polymerase amplification (RPA assay for rapid detection of Fno was developed and validated. The RPA reaction was performed at a constant temperature of 42°C for 20 min. The RPA assay was performed using a quantitative plasmid standard containing a unique Fno gene sequence. Validation of the assay was performed not only by using DNA from Fno, closely related Francisella species and other common bacterial pathogens in tilapia farms, but also by screening 78 Nile tilapia and 5 water samples. All results were compared with those obtained by previously established real-time qPCR. The developed RPA showed high specificity in detection of Fno with no cross-detection of either the closely related Francisella spp. or the other tested bacteria. The Fno-RPA performance was highly comparable to the published qPCR with detection limits at 15 and 11 DNA molecules detected, respectively. The RPA gave quicker results in approximately 6 min in contrast to the qPCR that needed about 90 min to reach the same detection limit, taking only 2.7-3 min to determine Fno in clinical samples. Moreover, RPA was more tolerant to reaction inhibitors than qPCR when tested with field samples. The fast reaction, simplicity, cost-effectiveness, sensitivity and specificity make the RPA an attractive diagnostic tool that will contribute to controlling the