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Sample records for fagus grandifolia subsp

  1. Formation of cis-coniferin in cell-free extracts of Fagus grandifolia Ehrh bark

    SciTech Connect

    Yamamoto, Etsuo; Inciong, M.E.J.; Davin, L.B.; Lewis, N.G. )

    1990-09-01

    American beech (Fagus grandifolia Ehrh) bark exclusively accumulates cis-monolignols and their glucosidic conjugates; no evidence for the accumulation of trans-monolignols has been found. The glucosyltransferase from this source exhibits a very unusual substrate specificity for cis, and not trans, monolignols. This is further evidence that cis monolignols are involved in lignin formation in these plant tissues. Preliminary evidence for the existence of a novel trans-cis monolignol isomerase was obtained, in agreement with our contention that this isomerization is not photochemically mediated.

  2. Formation of cis-coniferin in cell-free extracts of Fagus grandifolia Ehrh bark

    NASA Technical Reports Server (NTRS)

    Yamamoto, E.; Inciong, E. J.; Davin, L. B.; Lewis, N. G.

    1990-01-01

    American beech (Fagus grandifolia Ehrh) bark exclusively accumulates cis-monolignols and their glucosidic conjugates; no evidence for the accumulation of trans-monolignols has been found. The glucosyltransferase from this source exhibits a very unusual substrate specificity for cis, and not trans, monolignols. This is further evidence that cis monolignols are involved in lignin formation in these plant tissues. Preliminary evidence for the existence of a novel trans-cis monolignol isomerase was obtained, in agreement with our contention that this isomerization is not photochemically mediated.

  3. Exclusive accumulation of Z-isomers of monolignols and their glucosides in bark of Fagus grandifolia

    NASA Technical Reports Server (NTRS)

    Lewis, N. G.; Inciong, E. J.; Ohashi, H.; Towers, G. H.; Yamamoto, E.

    1988-01-01

    In addition to Z-coniferyl and Z-sinapyl alcohols, bark extracts of Fagus grandifolia also contain significant amounts of the glucosides, Z-coniferin, Z-isoconiferin (previously called faguside) and Z-syringin. The corresponding E-isomers of these glucosides do not accumulate to a detectable level. The accumulation of the Z-isomers suggests that either they are not lignin precursors or that they are reservoirs of monolignols for subsequent lignin biosynthesis; it is not possible to distinguish between these alternatives. The co-occurrence of Z-coniferin and Z-isoconiferin demonstrate that glucosylation of monolignols can occur at either the phenolic or the allylic hydroxyl groups.

  4. Utilizing pigment-producing fungi to add commercial value to American beech (Fagus grandifolia).

    PubMed

    Robinson, Sara C; Tudor, Daniela; Cooper, Paul A

    2012-02-01

    American beech (Fagus grandifolia) is an abundant, underutilized tree in certain areas of North America, and methods to increase its market value are of considerable interest. This research utilized pigment-producing fungi to induce color in American beech to potentially establish its use as a decorative wood. Wood samples were inoculated with Trametes versicolor, Xylaria polymorpha, Inonotus hispidus, and Arthrographis cuboidea to induce fungal pigmentation. Black pigmentation (T. versicolor, X. polymorpha, I. hispidus) was sporadic, occurred primarily on the surfaces of the heartwood, but not internally. Pink pigmentation (A. cuboidea) occurred throughout all of the tested beech samples, but was difficult to see in the heartwood due to the darker color of the wood. To increase the visibility of the pink stain, beech blocks were pretreated with T. versicolor for 4 weeks before being inoculated with A. cuboidea. This method significantly increased the saturation of the pink stain on both beech heartwood and sapwood, creating coloration similar to that found on sugar maple. This value-adding process should be particularly effective for small-scale wood pigmentation, and should help establish a market for this currently underutilized wood species. PMID:21931972

  5. Do Interspecific Differences in Sapling Growth Traits Contribute to the Co-dominance of Acer saccharum and Fagus grandifolia?

    PubMed Central

    Takahashi, Koichi; Lechowicz, Martin J.

    2008-01-01

    Background and Aims Acer saccharum and Fagus grandifolia are among the most dominant late-successional tree species in North America. The influence of sapling growth responses to canopy gaps on the co-dominance of the two species in an old-growth forest in southern Quebec, Canada was examined. Two predictions were evaluated: (a) F. grandifolia is more shade tolerant than A. saccharum due to greater sapling leaf area and net production per sapling in closed-canopy conditions; and (b) the height growth rate of A. saccharum in canopy gaps is greater than that of F. grandifolia due to increased net production per sapling. Methods Sapling crown allometry, net production and height growth rates were compared between and within the two species in closed canopy vs. canopy gaps. Standardized major axis regression was used to analyse differences in crown allometry. Key Results F. grandifolia had greater crown projection, sapling leaf area and net production rate per sapling than A. saccharum in closed-canopy conditions. In response to canopy gaps, net production per sapling increased to the same degree in both species. The net production per sapling of F. grandifolia thus was much greater than that of A. saccharum in both canopy gap and closed-canopy conditions. The height growth rate of both species increased in canopy gaps, but the degree of increase was greater in F. grandifolia than in A. saccharum. Conclusions F. grandifolia regenerated more successfully than A. saccharum in both closed-canopy conditions and canopy gaps, which indicates that the co-dominance of the two species cannot be maintained simply by interspecific differences in shade tolerance and growth in gaps. Previous research showed that although Fagus and Acer shared dominance at this site, their relative dominance shifted with edaphic conditions. This suggests that the widespread co-dominance of the two species in eastern North American forests is maintained by the joint influence of canopy disturbance and species-specific responses to the heterogeneity of moisture and fertility regimes within forested landscapes. PMID:17942590

  6. Two Lactarius species associated with a relict Fagus grandifolia var. mexicana population in a Mexican montane cloud forest.

    PubMed

    Montoya, L; Haug, I; Bandala, V M

    2010-01-01

    Ectomycorrhizal (EM) fleshy fungi are being monitored in a population of Fagus grandifolia var. mexicana persisting in a montane cloud forest refuge on a volcano in a subtropical region of central Veracruz (eastern Mexico). The population of Fagus studied represents one of the 10 recognized forest fragments still housing this tree genus in Mexico. This is the first attempt to document EM fungi associated with this tree species in Mexico. We present evidence of the ectomycorrhizal symbiosis for Lactarius badiopallescens and L. cinereus with this endemic tree. Species identification of Lactarius on Fagus grandifolia var. mexicana was based on the comparison of DNAsequences (ITS rDNA) of spatiotemporally co-occurring basidiomes and EM root tips. The host of the EM tips was identified by comparison of the large subunit of the ribulose-bisphosphate carboxylase gene (rbcL). The occurrence of Lactarius badiopallescens and L. cinereus populations in the area of study represent the southernmost record known to date of these two species in North America and are new for the Neotropical Lactarius mycota. Descriptions coupled with illustrations of macro- and micromorphological features of basidiomes as well as photographs of ectomycorrhizas are presented. PMID:20120238

  7. Morphological and molecular identification of the ectomycorrhizal association of Lactarius fumosibrunneus and Fagus grandifolia var. mexicana trees in eastern Mexico.

    PubMed

    Garay-Serrano, Edith; Bandala, Victor Manuel; Montoya, Leticia

    2012-11-01

    A population of Fagus grandifolia var. mexicana (covering ca. 4.7 ha) is established in a montane cloud forest refuge at Acatlan Volcano in eastern Mexico (Veracruz State), and it represents one of only ten populations of this species known to occur in the country (each stand covers ca. 2-35 ha in extension) and one of the southernmost in the continent. Sporocarps of several ectomycorrhizal macrofungi have been observed in the area, and among them, individuals of the genus Lactarius are common in the forest. However, the morphological and molecular characterization of ectomycorrhizae is still in development. Currently, two species of Lactarius have been previously documented in the area. Through the phylogenetic analysis of the internal transcribed spacer (ITS) region from basidiomes and ectomycorrhizae, we identified the Lactarius fumosibrunneus ectomycorrhiza. The host, F. grandifolia var. mexicana, was determined comparing the amplified ITS sequence from ectomycorrhizal root tips in the GenBank database with Basic Local Alignment Search Tool. The mycorrhizal system of L. fumosibrunneus is monopodial-pyramidal, characterized by its shiny, white to silver and pruinose surface, secreting a white latex when damaged, composed of three plectenchymatous mantle layers, with diverticulated terminal elements at the outer mantle. It lacks emanating hyphae, rhizomorphs, and sclerotia. A detailed morphological and anatomical description, illustrations, and photographs of the ectomycorrhiza are presented. The comparison of L. fumosibrunneus and other Lactarius belonging to subgenus Plinthogalus is presented. PMID:22402818

  8. Unexpected presence of Fagus orientalis complex in Italy as inferred from 45,000-year-old DNA pollen samples from Venice lagoon

    PubMed Central

    Paffetti, Donatella; Vettori, Cristina; Caramelli, David; Vernesi, Cristiano; Lari, Martina; Paganelli, Arturo; Paule, Ladislav; Giannini, Raffaello

    2007-01-01

    Background Phylogeographic analyses on the Western Euroasiatic Fagus taxa (F. orientalis, F. sylvatica, F. taurica and F. moesiaca) is available, however, the subdivision of Fagus spp. is unresolved and there is no consensus on the phylogeny and on the identification (both with morphological than molecular markers) of Fagus Eurasiatic taxa. For the first time molecular analyses of ancient pollen, dated at least 45,000 years ago, were used in combination with the phylogeny analysis on current species, to identify the Fagus spp. present during the Last Interglacial period in Italy. In this work we aim at testing if the trnL-trnF chloroplast DNA (cpDNA) region, that has been previously proved efficient in discriminating different Quercus taxa, can be employed in distinguishing the Fagus species and in identifying the ancient pollen. Results 86 populations from 4 Western Euroasistic taxa were sampled, and sequenced for the trnL-trnF region to verify the efficiency of this cpDNA region in identifying the Fagus spp.. Furthermore, Fagus crenata (2 populations), Fagus grandifolia (2 populations), Fagus japonica, Fagus hayatae, Quercus species and Castanea species were analysed to better resolve the phylogenetic inference. Our results show that this cpDNA region harbour some informative sites that allow to infer relationships among the species within the Fagaceae family. In particular, few specific and fixed mutations were able to discriminate and identify all the different Fagus species. Considering a short fragment of 176 base pairs within the trnL intron, 2 transversions were found able in distinguishing the F. orientalis complex taxa (F. orientalis, F. taurica and F. moesiaca) from the remaining Fagus spp. (F. sylvatica, F. japonica, F. hayataea, F. crenata and F. grandifolia). This permits to analyse this fragment also in ancient samples, where DNA is usually highly degraded. The sequences data indicate that the DNA recovered from ancient pollen belongs to the F. orientalis complex since it displays the informative sites characteristic of this complex. Conclusion The ancient DNA sequences demonstrate for the first time that, in contrast to current knowledge based on palynological and macrofossil data, the F. orientalis complex was already present during the Tyrrhenian period in what is now the Venice lagoon (Italy). This is a new and important insight considering that nowadays West Europe is not the natural area of Fagus orientalis complex, and up to now nobody has hypothesized the presence during the Last Interglacial period of F. orientalis complex in Italy. PMID:17767734

  9. Comparisons of protein profiles of beech bark disease resistant and susceptible American beech (Fagus grandifolia)

    PubMed Central

    2013-01-01

    Background Beech bark disease is an insect-fungus complex that damages and often kills American beech trees and has major ecological and economic impacts on forests of the northeastern United States and southeastern Canadian forests. The disease begins when exotic beech scale insects feed on the bark of trees, and is followed by infection of damaged bark tissues by one of the Neonectria species of fungi. Proteomic analysis was conducted of beech bark proteins from diseased trees and healthy trees in areas heavily infested with beech bark disease. All of the diseased trees had signs of Neonectria infection such as cankers or fruiting bodies. In previous tests reported elsewhere, all of the diseased trees were demonstrated to be susceptible to the scale insect and all of the healthy trees were demonstrated to be resistant to the scale insect. Sixteen trees were sampled from eight geographically isolated stands, the sample consisting of 10 healthy (scale-resistant) and 6 diseased/infested (scale-susceptible) trees. Results Proteins were extracted from each tree and analysed in triplicate by isoelectric focusing followed by denaturing gel electrophoresis. Gels were stained and protein spots identified and intensity quantified, then a statistical model was fit to identify significant differences between trees. A subset of BBD differential proteins were analysed by mass spectrometry and matched to known protein sequences for identification. Identified proteins had homology to stress, insect, and pathogen related proteins in other plant systems. Protein spots significantly different in diseased and healthy trees having no stand or disease-by-stand interaction effects were identified. Conclusions Further study of these proteins should help to understand processes critical to resistance to beech bark disease and to develop biomarkers for use in tree breeding programs and for the selection of resistant trees prior to or in early stages of BBD development in stands. Early identification of resistant trees (prior to the full disease development in an area) will allow forest management through the removal of susceptible trees and their root-sprouts prior to the onset of disease, allowing management and mitigation of costs, economic impact, and impacts on ecological systems and services. PMID:23317283

  10. Antimalarial diterpenoid dimers of a new carbon skeleton from Aphanamixis grandifolia.

    PubMed

    Zhang, Hua; Liu, Jia; Gan, Li-She; Dalal, Seema; Cassera, Maria B; Yue, Jian-Min

    2016-01-21

    Chemical investigation into the minor constituents of Aphanamixis grandifolia yielded three new diterpenoid dimers, aphadilactones E-G (1-3) featuring a new carbon skeleton. Their structures and absolute configurations were fully established by comprehensive spectroscopic data analysis and ECD calculation. Discovery of another two new dimers (4 and 5) suggested the structure of recently reported aphanamene A to be re-investigated. Compounds 1-5 showed moderate antimalarial activities with low micromolar IC50 values. PMID:26615760

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

    EPA Science Inventory

    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...

  12. Molecular Characterization of Copper Resistance Genes from Xanthomonas citri subsp. citri and Xanthomonas alfalfae subsp. citrumelonis?

    PubMed Central

    Behlau, Franklin; Canteros, Blanca I.; Minsavage, Gerald V.; Jones, Jeffrey B.; Graham, James H.

    2011-01-01

    Copper sprays have been widely used for control of endemic citrus canker caused by Xanthomonas citri subsp. citri in citrus-growing areas for more than 2 decades. Xanthomonas alfalfae subsp. citrumelonis populations were also exposed to frequent sprays of copper for several years as a protective measure against citrus bacterial spot (CBS) in Florida citrus nurseries. Long-term use of these bactericides has led to the development of copper-resistant (Cur) strains in both X. citri subsp. citri and X. alfalfae subsp. citrumelonis, resulting in a reduction of disease control. The objectives of this study were to characterize for the first time the genetics of copper resistance in X. citri subsp. citri and X. alfalfae subsp. citrumelonis and to compare these organisms to other Cur bacteria. Copper resistance determinants from X. citri subsp. citri strain A44(pXccCu2) from Argentina and X. alfalfae subsp. citrumelonis strain 1381(pXacCu2) from Florida were cloned and sequenced. Open reading frames (ORFs) related to the genes copL, copA, copB, copM, copG, copC, copD, and copF were identified in X. citri subsp. citri A44. The same ORFs, except copC and copD, were also present in X. alfalfae subsp. citrumelonis 1381. Transposon mutagenesis of the cloned copper resistance determinants in pXccCu2 revealed that copper resistance in X. citri subsp. citri strain A44 is mostly due to copL, copA, and copB, which are the genes in the cloned cluster with the highest nucleotide homology (?92%) among different Cur bacteria. PMID:21515725

  13. Common Genomic Features of Campylobacter jejuni subsp. doylei Strains Distinguish Them from C. jejuni subsp. jejuni

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

  14. Genome Sequences of the Listeria ivanovii subsp. ivanovii Type Strain and Two Listeria ivanovii subsp. londoniensis Strains

    PubMed Central

    Hupfeld, Mario; Fouts, Derrick E.; Loessner, Martin J.

    2015-01-01

    We present the complete genomes of Listeria ivanovii subsp. ivanovii WSLC 3010 (ATCC 19119T), Listeria ivanovii subsp. londoniensis WSLC 30151 (SLCC 8854), and Listeria ivanovii subsp. londoniensis WSLC 30167 (SLCC 6032), representing the type strain of the species and two strains of the same serovar but different properties, respectively. PMID:25614561

  15. Campylobacter fetus subsp. testudinum subsp. nov., isolated from humans and reptiles.

    PubMed

    Fitzgerald, Collette; Tu, Zheng Chao; Patrick, Mary; Stiles, Tracy; Lawson, Andy J; Santovenia, Monica; Gilbert, Maarten J; van Bergen, Marcel; Joyce, Kevin; Pruckler, Janet; Stroika, Steven; Duim, Birgitta; Miller, William G; Loparev, Vladimir; Sinnige, Jan C; Fields, Patricia I; Tauxe, Robert V; Blaser, Martin J; Wagenaar, Jaap A

    2014-09-01

    A polyphasic study was undertaken to determine the taxonomic position of 13 Campylobacter fetus-like strains from humans (n?=?8) and reptiles (n?=?5). The results of matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS and genomic data from sap analysis, 16S rRNA gene and hsp60 sequence comparison, pulsed-field gel electrophoresis, amplified fragment length polymorphism analysis, DNA-DNA hybridization and whole genome sequencing demonstrated that these strains are closely related to C. fetus but clearly differentiated from recognized subspecies of C. fetus. Therefore, this unique cluster of 13 strains represents a novel subspecies within the species C. fetus, for which the name Campylobacter fetus subsp. testudinum subsp. nov. is proposed, with strain 03-427(T) (?=?ATCC BAA-2539(T)?=?LMG 27499(T)) as the type strain. Although this novel taxon could not be differentiated from C. fetus subsp. fetus and C. fetus subsp. venerealis using conventional phenotypic tests, MALDI-TOF MS revealed the presence of multiple phenotypic biomarkers which distinguish Campylobacter fetus subsp. testudinum subsp. nov. from recognized subspecies of C. fetus. PMID:24899653

  16. Growth of Fagus in transition zones of forest and soil on the western slope of Mt. Chokai, northern Japan

    NASA Astrophysics Data System (ADS)

    Kato, S.; Watanabe, M.

    2012-04-01

    A wide transition zone for forest structure is expected to distribute on the gentle slope of western side of Mt. Chokai ,Yamagata prefecture, northern Japan (N39° 05'57", E140°02'55"). The annual mean temperature and total precipitation at summit (2,059 m asl.) are 0.5° C and 3,285mm, respectively. The parent materials of the soils are weathered Andesite associated with non-tephric loess deposits transported from continental China. Representative sites were selected in forests of Quercus mongolica and Fagus crenata to examine characteristics of transition zones of vegetation and soil in the western slope of Mt. Chokai with concern on the growth of Fagus in transition zones. Surveys on vegetation profile and projection diagram of canopy for each site (10-10m plots) were carried out in 7 sites selected along altitudinal sequence on the western slope of Mt. Chokai; Ch1-7: 550-1,100m asl.. Growth rate of Fagus was estimated by the measurement of tree rings from increment core samples. Timber volume of Fagus at each point was calculated based on diameter of breast height; DBH as an indicator of tree biomass. Soil profiles were observed at the above 7 sites and soil samples were collected from each horizon. As for soil analyses, soil pH (H2O, KCl, NaF) values were measured by the glass electrode method in the suspension mixture of soil with a 2.5 times volume of H2O or 1N KCl and 50 times volume of 4% NaF. Pyrophosphate, acid oxalate and dithionite-citrate extractable Al (Alp, Alo, Ald), Fe (Feo, Fed) and Si (Sio, Sid) were measured by ICP-AES. The content of exchangeable Al (AlEX) was obtained by titration of extract with 1N KCl. Sclerotia formed by species of Cenococcum, ectomycorrhizal fungi, were collected for grains of diameter larger than 0.5mm from wet samples. Sclerotia content was obtained by weight (mg g-1 soil). Due to intensive base leaching under extremely high precipitation and the mineralogical properties, Ah and Ae horizons of all profiles had low soil pH values (3.4-4.6 in H2O) and a high content of exchangeable Al (0.20-1.09g kg-1 soil), which could restrict nutrient uptake and growth of trees. From observation of forest structures of each site, transition zone was approximately 750m by horizontal distance including Ch3-5 (710-780m asl.). The DBH of Fagus was more than 45cm at Ch3,4, and the timber volume of Fagus was the largest at Ch3, followed by Ch4, and the smallest at Ch7, followed by Ch6. Among the entire 7 sites, the growth rate of Fagus was the largest at Ch3 where soil pH value was above 4.0 and exchangeable Al content was relatively low. The oldest and highest Fagus was observed at Ch4, where content of exchangeable Al was high (1.0 g kg-1) . While, Fagus at Ch3,5 and 7 had almost same age. Sclerotia content in soils was much larger in transition zone (Ch3,4) than in podzolic soils (Ch6,7) beneath the dwarfed Fagus forest. The activity of mycorrhizae was suggested to be one of the important factors for the growth of Fagus in transition zone.

  17. Fine roots in stands of Fagus sylvatica and Picea abies along a gradient of soil acidification.

    PubMed

    Braun, Sabine; Cantaluppi, Leonardo; Flückiger, Walter

    2005-10-01

    Root length of naturally grown young beech trees (Fagus sylvatica L.) was investigated in 26 forest plots of differing base saturation and nitrogen deposition. The relative length of finest roots (<0.25 mm) was found to decrease in soils with low base saturation. A similar reduction of finest roots in plots with high nitrogen deposition was masked by the effect of base saturation. The formation of adventitious roots was enhanced in acidic soils. The analysis of 128 soil profiles for fine roots of all species present in stands of either Fagus sylvatica L., Picea abies [Karst.] L. or both showed a decreased rooting depth in soils with < or =20% base saturation and in hydromorphic soils. For base rich, well drained soils an average rooting depth of 108 cm was found. This decreased by 28 cm on acidic, well drained soils. The results suggest an effect of the current soil acidification in Switzerland and possibly also of nitrogen deposition on the fine root systems of forest trees. PMID:15964116

  18. Frequency of inversions affects senescence phenology of Acer pseudoplatanus and Fagus sylvatica.

    PubMed

    Schuster, Christina; Kirchner, Manfred; Jakobi, Gert; Menzel, Annette

    2014-05-01

    In mountainous regions, inversion situations with cold-air pools in the valleys occur frequently, especially in fall and winter. With the accumulation of inversion days, trees in lower elevations experience lower temperature sums than those in middle elevations. In a two-year observational study, deciduous trees, such as Acer pseudoplatanus and Fagus sylvatica, on altitudinal transects responded in their fall leaf senescence phenology. Phenological phases were advanced and senescence duration was shortened by the cold temperatures in the valley. This effect was more distinct for late phases than for early phases since they experienced more inversion days. The higher the inversion frequency, the stronger the signal was. Acer pseudoplatanus proved to be more sensitive to cold temperatures compared to Fagus sylvatica. We conclude that cold-air pools have a considerable impact on the vegetation period of deciduous trees. Considering this effect, trees in the mid hillside slopes gain advantages compared to lower elevations. Our findings will help to improve knowledge about ecological drivers and responses in mountainous forest ecosystems. PMID:23912394

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    PubMed

    Švec, Pavel; De Bel, Annelies; Sedlá?ek, Ivo; Petráš, Petr; Gelbí?ová, Tereza; ?ernohlávková, Jitka; Mašlan?ová, Ivana; Cnockaert, Margo; Varbanovová, Ivana; Echahidi, Fedoua; Vandamme, Peter; Pantu?ek, 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)). PMID:25829332

  1. Complete genome sequence of Bifidobacterium animalis subsp. lactis BLC1.

    PubMed

    Bottacini, Francesca; Dal Bello, Fabio; Turroni, Francesca; Milani, Christian; Duranti, Sabrina; Foroni, Elena; Viappiani, Alice; Strati, Francesco; Mora, Diego; van Sinderen, Douwe; Ventura, Marco

    2011-11-01

    Bifidobacterium animalis subsp. lactis BLC1 is a probiotic bacterium that is widely exploited by food industries as the active ingredient of various functional foods. Here we report the complete genome sequence of B. animalis subsp. lactis BLC1, which is expected to provide insights into the biology of this health-promoting microorganism and improve our understanding of its phylogenetic relatedness with other members of the B. animalis subsp. lactis taxon. PMID:22038957

  2. Fast wood decay in a mountain Mediterranean area having Fagus sylvatica forests

    NASA Astrophysics Data System (ADS)

    Fravolini, Giulia; Egli, Markus; Cherubini, Paolo; Tognetti, Roberto; Lombardi, Fabio; Marchetti, Marco

    2015-04-01

    Deadwood and litter act as important linkages between recent productivity and current community, and ecosystem processes. The increasing interest in the quantity and properties of coarse woody debris (CWD) and litter is relevant both to maintaining biodiversity and to global C dynamics. Mountain and Mediterranean areas, furthermore, are considered to be especially sensitive to changing environmental conditions. Consequently, a need exists to understand more in detail the interplay between soils, forests, deadwood and climate in general and in particular in mountain Mediterranean areas such as the Appenine. Due to the fact that linkages between climate, coarse woody decay and soils in mountain Mediterranean areas are only poorly understood, we aimed at investigating the decay mechanism of Fagus silvatica as a function of altitude and exposure. Furthermore, the effects of exposure on the decay dynamics of dead wood and soils were compared along a altitudinal sequence in an Appenine mountain forest (Majella Mountain). Ten sites, five of which having north and the other 5 having south exposure, were investigated, ranging from 1000 m to 1650 m asl. All sites have a Fagus sylvatica forest. In addition to this, experimental plots were installed at each site. In May 2014 standardised wood blocks (5 x 5 x 2 cm) of local Fagus sylvatica were placed at each site inside PVC tubes ('mesocosms') that was filled with undisturbed soil material. The sampling design foresees that three replicates of such mesocosms per site will be sampled after 8 , 16, 52 and 104 weeks. After 8 weeks three tubes were removed from the sites (sampled soil and dead wood blocks) and the wood blocks analysed for cellulose, lignin and density. At each site, three cores were taken to analyse soil properties. The soil cores were subdivided in 0 - 5, 5 - 10 and 10 - 15 cm depth and measured for organic carbon, carbonates and pH. In addition, the humus forms at each site were determined. Already after 8 weeks, the lignin content of the dead wood increased compared to the fresh wood. The cellulose analysis, however, showed a slight decreasing trend (not always statistically significant), both at the north and the south exposure. Especially at the highest altitudes this decreasing trend was best expressed. The highest sites are the coolest and also have enough moisture of during the dry summer months. Together with the relatively warm conditions, decay processes seem to be enhanced there. The humus forms nicely represented the climatic conditions with predominately Mull at higher sites and particularly at north-facing sites and Amphi at lower and warmer sites. Consequently, a fast wood decay also correlates with the humus form Mull.

  3. Antibacterial Activity of Alkyl Gallates against Xanthomonas citri subsp. citri

    PubMed Central

    Silva, I. C.; Regasini, L. O.; Petrônio, M. S.; Silva, D. H. S.; Bolzani, V. S.; Belasque, J.; Sacramento, L. V. S.

    2013-01-01

    The plant-pathogenic bacterium Xanthomonas citri subsp. citri is the causal agent of Asiatic citrus canker, a serious disease that affects all the cultivars of citrus in subtropical citrus-producing areas worldwide. There is no curative treatment for citrus canker; thus, the eradication of infected plants constitutes the only effective control of the spread of X. citri subsp. citri. Since the eradication program in the state of São Paulo, Brazil, is under threat, there is a clear risk of X. citri subsp. citri becoming endemic in the main orange-producing area in the world. Here we evaluated the potential use of alkyl gallates to prevent X. citri subsp. citri growth. These esters displayed a potent anti-X. citri subsp. citri activity similar to that of kanamycin (positive control), as evaluated by the resazurin microtiter assay (REMA). The treatment of X. citri subsp. citri cells with these compounds induced altered cell morphology, and investigations of the possible intracellular targets using X. citri subsp. citri strains labeled for the septum and centromere pointed to a common target involved in chromosome segregation and cell division. Finally, the artificial inoculation of citrus with X. citri subsp. citri cells pretreated with alkyl gallates showed that the bacterium loses the ability to colonize its host, which indicates the potential of these esters to protect citrus plants against X. citri subsp. citri infection. PMID:23104804

  4. Do variations in leaf phenology affect radial growth variations in Fagus sylvatica?

    NASA Astrophysics Data System (ADS)

    ?ufar, Katarina; De Luis, Martin; Prislan, Peter; Gri?ar, Jožica; ?repinšek, Zalika; Merela, Maks; Kajfež-Bogataj, Lu?ka

    2015-08-01

    We used a dendrochronological and leaf phenology network of European beech ( Fagus sylvatica) in Slovenia, a transitional area between Mediterranean, Alpine and continental climatic regimes, for the period 1955-2007 to test whether year to year variations in leaf unfolding and canopy duration (i.e. time between leaf unfolding and colouring) influence radial growth (annual xylem production and tree ring widths) and if such influences are more pronounced at higher altitudes. We showed that variability in leaf phenology has no significant effect on variations in radial growth. The results are consistent in the entire region, irrespective of the climatic regime or altitude, although previous studies have shown that leaf phenology and tree ring variation depend on altitude. The lack of relationship between year to year variability in leaf phenology and radial growth may suggest that earlier leaf unfolding—as observed in a previous study—probably does not cause increased tree growth rates in beech in Slovenia.

  5. Drought-Adaptation Potential in Fagus sylvatica: Linking Moisture Availability with Genetic Diversity and Dendrochronology

    PubMed Central

    Pluess, Andrea R.; Weber, Pascale

    2012-01-01

    Background Microevolution is essential for species persistence especially under anticipated climate change scenarios. Species distribution projection models suggested that the dominant tree species of lowland forests in Switzerland, European beech (Fagus sylvatica L.), might disappear from most areas due to expected longer dry periods. However, if genotypes at the moisture boundary of the species climatic envelope are adapted to lower moisture availability, they can serve as seed source for the continuation of beech forests under changing climates. Methodology/Principal Findings With an AFLP genome scan approach, we studied neutral and potentially adaptive genetic variation in Fagus sylvatica in three regions containing a dry and a mesic site each (nind.?=?241, nmarkers?=?517). We linked this dataset with dendrochronological growth measures and local moisture availabilities based on precipitation and soil characteristics. Genetic diversity decreased slightly at dry sites. Overall genetic differentiation was low (Fst?=?0.028) and Bayesian cluster analysis grouped all populations together suggesting high (historical) gene flow. The Bayesian outlier analyses indicated 13 markers with three markers differing between all dry and mesic sites and the others between the contrasting sites within individual regions. A total of 41 markers, including seven outlier loci, changed their frequency with local moisture availability. Tree height and median basal growth increments were reduced at dry sites, but marker presence/absence was not related to dendrochronological characteristics. Conclusion and Their Significance The outlier alleles and the makers with changing frequencies in relation to moisture availability indicate microevolutionary processes occurring within short geographic distances. The general genetic similarity among sites suggests that ‘preadaptive’ genes can easily spread across the landscape. Yet, due to the long live span of trees, fostering saplings originating from dry sites and grown within mesic sites might increase resistance of beech forests during the anticipated longer dry periods. PMID:22448260

  6. Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation.

    PubMed

    Genersch, Elke; Forsgren, Eva; Pentikäinen, Jaana; Ashiralieva, Ainura; Rauch, Sandra; Kilwinski, Jochen; Fries, Ingemar

    2006-03-01

    A polyphasic taxonomic study of the two subspecies of Paenibacillus larvae, Paenibacillus larvae subsp. larvae and Paenibacillus larvae subsp. pulvifaciens, supported the reclassification of the subspecies into one species, Paenibacillus larvae, without subspecies separation. Our conclusions are based on the analysis of six reference strains of P. larvae subsp. pulvifaciens and three reference strains and 44 field isolates of P. larvae. subsp. larvae. The latter originated from brood or honey of clinically diseased honey bee colonies or from honey of both clinically diseased and asymptomatic colonies from Sweden, Finland and Germany. Colony and spore morphology, as well as the metabolism of mannitol and salicin, did not allow a clear identification of the two subspecies and SDS-PAGE of whole-cell proteins did not support the subspecies differentiation. For genomic fingerprinting, repetitive element-PCR fingerprinting using ERIC primers and PFGE of bacterial DNA were performed. The latter method is a high-resolution DNA fingerprinting method proven to be superior to most other methods for biochemical and molecular typing and has not previously been used to characterize P. larvae. ERIC-PCR identified four different genotypes, while PFGE revealed two main clusters. One cluster included most of the P. larvae subsp. larvae field isolates, as well as all P. larvae subsp. pulvifaciens reference strains. The other cluster comprised the pigmented variants of P. larvae subsp. larvae. 16S rRNA gene sequences were determined for some strains. Finally, exposure bioassays demonstrated that reference strains of P. larvae subsp. pulvifaciens were pathogenic for honey bee larvae, producing symptoms similar to reference strains of P. larvae subsp. larvae. In comparison with the type strain for P. larvae subsp. larvae, ATCC 9545T, the P. larvae subsp. pulvifaciens strains tested were even more virulent, since they showed a shorter LT100. An emended description of the species is given. PMID:16514018

  7. Mycobacterium abscessus subsp abscessus lung disease: 'trouble ahead, trouble behind…'.

    PubMed

    Griffith, David E

    2014-01-01

    Mycobacterium abscessus subsp abscessus is the most common respiratory pathogen among the rapidly growing non-tuberculous mycobacteria (NTM) and is also the most feared due to its well-deserved reputation for being refractory to antibiotic therapy. M. abscessus subsp abscessus has multiple innate antibiotic resistance mechanisms, but the most important one described so far is an inducible erythromycin methylase (erm) gene. M. abscessus subsp abscessus isolates may appear macrolide susceptible on initial in vitro testing but become macrolide resistant after exposure to macrolide. It is therefore very important to test clinically significant M. abscessus subsp abscessus isolates for erm gene activity. Remarkably, controversy still exists about the taxonomy and nomenclature of M. abscessus subspecies including subsp abscessus, subsp massiliense and subsp bolletii. Identification of these subspecies is not moot as M. abscessus subsp massiliense does not have an active erm gene resulting in both in vitro and in vivo susceptibility to macrolide. It is imperative from the clinician's perspective that mycobacterial laboratories correctly and rapidly identify M. abscessus to the subspecies level. Unfortunately, there are no reliably or predictably effective treatment regimens for M. abscessus subsp abscessus and better, more effective antimicrobial agents are badly needed. Surgical resection of involved lung tissue as an adjunct to antibiotic therapy is beneficial in selected patients but cannot be broadly applied. Overall, M. abscessus subsp abscessus remains a formidable respiratory mycobacterial pathogen, one that we are only beginning to understand microbiologically and one that as yet consistently evades our best efforts at successful therapeutic outcomes. 'trouble ahead, trouble behind, and you know that notion just crossed my mind'.Casey Jones, Grateful Dead (1970). PMID:25580261

  8. Proposal to rename Carnobacterium inhibens as Carnobacterium inhibens subsp. inhibens subsp. nov. and description of Carnobacterium inhibens subsp. gilichinskyi subsp. nov., a psychrotolerant bacterium isolated from Siberian permafrost.

    PubMed

    Nicholson, Wayne L; Zhalnina, Kateryna; de Oliveira, Rafael R; Triplett, Eric W

    2015-02-01

    A novel, psychrotolerant facultative anaerobe, strain WN1359(T), was isolated from a permafrost borehole sample collected at the right bank of the Kolyma River in Siberia, Russia. Gram-positive-staining, non-motile, rod-shaped cells were observed with sizes of 1-2 µm long and 0.4-0.5 µm wide. Growth occurred in the range of pH 5.8-9.0 with optimal growth at pH 7.8-8.6 (pH optimum 8.2). The novel isolate grew at temperatures from 0-37 °C and optimal growth occurred at 25 °C. The novel isolate does not require NaCl; growth was observed between 0 and 8.8 % (1.5 M) NaCl with optimal growth at 0.5 % (w/v) NaCl. The isolate was a catalase-negative, facultatively anaerobic chemo-organoheterotroph that used sugars but not several single amino acids or dipeptides as substrates. The major metabolic end-product was lactic acid in the ratio of 86 % l-lactate : 14 % d-lactate. Strain WN1359(T) was sensitive to ampicillin, chloramphenicol, fusidic acid, lincomycin, monocycline, rifampicin, rifamycin SV, spectinomycin, streptomycin, troleandomycin and vancomycin, and resistant to nalidixic acid and aztreonam. The fatty acid content was predominantly unsaturated (70.2 %), branched-chain unsaturated (11.7 %) and saturated (12.5 %). The DNA G+C content was 35.3 mol% by whole genome sequence analysis. 16S rRNA gene sequence analysis showed 98.7 % sequence identity between strain WN1359(T) and Carnobacterium inhibens. Genome relatedness was computed using both Genome-to-Genome Distance Analysis (GGDA) and Average Nucleotide Identity (ANI), which both strongly supported strain WN1359(T) belonging to the species C. inhibens. On the basis of these results, the permafrost isolate WN1359(T) represents a novel subspecies of C. inhibens, for which the name Carnobacterium inhibens subsp. gilichinskyi subsp. nov. is proposed. The type strain is WN1359(T) (?= ATCC BAA-2557(T)?= DSM 27470(T)). The subspecies Carnobacterium inhibens subsp. inhibens subsp. nov. is created automatically. An emended description of C. inhibens is also provided. PMID:25392348

  9. A pyrosequencing insight into sprawling bacterial diversity and community dynamics in decaying deadwood logs of Fagus sylvatica and Picea abies

    PubMed Central

    Hoppe, Björn; Krger, Krüger; Kahl, Tiemo; Arnstadt, Tobias; Buscot, François; Bauhus, Jürgen; Wubet, Tesfaye

    2015-01-01

    Deadwood is an important biodiversity hotspot in forest ecosystems. While saproxylic insects and wood-inhabiting fungi have been studied extensively, little is known about deadwood-inhabiting bacteria. The study we present is among the first to compare bacterial diversity and community structure of deadwood under field conditions. We therefore compared deadwood logs of two temperate forest tree species Fagus sylvatica and Picea abies using 16S rDNA pyrosequencing to identify changes in bacterial diversity and community structure at different stages of decay in forest plots under different management regimes. Alphaproteobacteria, Acidobacteria and Actinobacteria were the dominant taxonomic groups in both tree species. There were no differences in bacterial OTU richness between deadwood of Fagus sylvatica and Picea abies. Bacteria from the order Rhizobiales became more abundant during the intermediate and advanced stages of decay, accounting for up to 25% of the entire bacterial community in such logs. The most dominant OTU was taxonomically assigned to the genus Methylovirgula, which was recently described in a woodblock experiment of Fagus sylvatica. Besides tree species we were able to demonstrate that deadwood physico-chemical properties, in particular remaining mass, relative wood moisture, pH, and C/N ratio serve as drivers of community composition of deadwood-inhabiting bacteria. PMID:25851097

  10. A pyrosequencing insight into sprawling bacterial diversity and community dynamics in decaying deadwood logs of Fagus sylvatica and Picea abies.

    PubMed

    Hoppe, Björn; Krger, Krüger; Kahl, Tiemo; Arnstadt, Tobias; Buscot, François; Bauhus, Jürgen; Wubet, Tesfaye

    2015-01-01

    Deadwood is an important biodiversity hotspot in forest ecosystems. While saproxylic insects and wood-inhabiting fungi have been studied extensively, little is known about deadwood-inhabiting bacteria. The study we present is among the first to compare bacterial diversity and community structure of deadwood under field conditions. We therefore compared deadwood logs of two temperate forest tree species Fagus sylvatica and Picea abies using 16S rDNA pyrosequencing to identify changes in bacterial diversity and community structure at different stages of decay in forest plots under different management regimes. Alphaproteobacteria, Acidobacteria and Actinobacteria were the dominant taxonomic groups in both tree species. There were no differences in bacterial OTU richness between deadwood of Fagus sylvatica and Picea abies. Bacteria from the order Rhizobiales became more abundant during the intermediate and advanced stages of decay, accounting for up to 25% of the entire bacterial community in such logs. The most dominant OTU was taxonomically assigned to the genus Methylovirgula, which was recently described in a woodblock experiment of Fagus sylvatica. Besides tree species we were able to demonstrate that deadwood physico-chemical properties, in particular remaining mass, relative wood moisture, pH, and C/N ratio serve as drivers of community composition of deadwood-inhabiting bacteria. PMID:25851097

  11. Variation in biogenic volatile organic compound emission pattern of Fagus sylvatica L. due to aphid infection

    NASA Astrophysics Data System (ADS)

    Joó, É.; Van Langenhove, H.; Šimpraga, M.; Steppe, K.; Amelynck, C.; Schoon, N.; Müller, J.-F.; Dewulf, J.

    2010-01-01

    Volatile organic compounds (VOCs) have been the focus of interest to understand atmospheric processes and their consequences in formation of ozone or aerosol particles; therefore, VOCs contribute to climate change. In this study, biogenic VOCs (BVOCs) emitted from Fagus sylvatica L. trees were measured in a dynamic enclosure system. In total 18 compounds were identified: 11 monoterpenes (MT), an oxygenated MT, a homoterpene (C 14H 18), 3 sesquiterpenes (SQT), isoprene and methyl salicylate. The frequency distribution of the compounds was tested to determine a relation with the presence of the aphid Phyllaphis fagi L. It was found that linalool, (E)-?-ocimene, ?-farnesene and a homoterpene identified as (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), were present in significantly more samples when infection was present on the trees. The observed emission spectrum from F. sylvatica L. shifted from MT to linalool, ?-farnesene, (E)-?-ocimene and DMNT due to the aphid infection. Sabinene was quantitatively the most prevalent compound in both, non-infected and infected samples. In the presence of aphids ?-farnesene and linalool became the second and third most important BVOC emitted. According to our investigation, the emission fingerprint is expected to be more complex than commonly presumed.

  12. Mycobacterium avium subsp. paratuberculosis in Veterinary Medicine

    PubMed Central

    Harris, N. Beth; Barletta, Raúl G.

    2001-01-01

    Mycobacterium avium subsp. paratuberculosis (basonym M. paratuberculosis) is the etiologic agent of a severe gastroenteritis in ruminants known as Johne's disease. Economic losses to the cattle industry in the United States are staggering, reaching $1.5 billion annually. A potential pathogenic role in humans in the etiology of Crohn's disease is under investigation. In this article, we review the epidemiology, pathogenesis, diagnostics, and disease control measures of this important veterinary pathogen. We emphasize molecular genetic aspects including the description of markers used for strain identification, diagnostics, and phylogenetic analysis. Recent important advances in the development of animal models and genetic systems to study M. paratuberculosis virulence determinants are also discussed. We conclude with proposals for the applications of these models and recombinant technology to the development of diagnostic, control, and therapeutic measures. PMID:11432810

  13. Functional characterization of a dehydrin protein from Fagus sylvatica seeds using experimental and in silico approaches.

    PubMed

    Kalemba, Ewa Marzena; Litkowiec, Monika

    2015-12-01

    A strong increase in the level of dehydrin/response ABA transcripts expression reported from the 14th week after flowering coincident with the accumulation of 26 and 44 kDa dehydrins in the embryonic axes of developing beech (Fagus sylvatica L.) seeds. Both transcript and protein levels were strongly correlated with maturation drying. These results suggest that the 44-kDa dehydrin protein is a putative dimer of dehydrin/response ABA protein migrating as a 26-kDa protein. Dehydrins and dehydrin-like proteins form large oligomeric complexes under native conditions and are shown as several spots differing in pI through isoelectrofocusing analyses. Detailed prediction of specific sites accessible for various post-translational modifications (PTMs) in the dehydrin/response ABA protein sequence revealed sites specific to acetylation, amidation, glycosylation, methylation, myristoylation, nitrosylation, O-linked ?-N-acetylglucosamination and Yin-O-Yang modification, palmitoylation, phosphorylation, sumoylation, sulfation, and ubiquitination. Thus, these results suggest that specific PTMs might play a role in switching dehydrin function or activity, water binding ability, protein-membrane interactions, transport and subcellular localization, interactions with targeted molecules, and protein stability. Despite the ability of two Cys residues to form a disulfide bond, -SH groups are likely not involved in dimer arrangement. His-rich regions and/or polyQ-tracts are potential candidates as spatial organization modulators. Dehydrin/response ABA protein is an intrinsically disordered protein containing low complexity regions. The lack of a fixed structure and exposition of amino acids on the surface of the protein structure enhances the accessibility to 40 predicted PTM sites, thereby facilitating dehydrin multifunctionality, which is discussed in the present study. PMID:26492132

  14. Potential Transmission Pathways of Streptococcus gallolyticus subsp. gallolyticus

    PubMed Central

    Dumke, Jessika; Hinse, Dennis; Vollmer, Tanja; Schulz, Jochen; Knabbe, Cornelius; Dreier, Jens

    2015-01-01

    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. PMID:25978355

  15. Changes in BVOC emission pattern from Fagus sylvatica L. measured by thermal desorber GC-MS

    NASA Astrophysics Data System (ADS)

    Joó, É.; van Langenhove, H.; Schietse, L.; Pokorska, O.; Šimpraga, M.; Steppe, K.; Demarcke, M.; Amelynck, C.; Schoon, N.; Müller, J.-F.; Samson, R.; Dewulf, J.

    2009-04-01

    Considerable attention has been focused on biogenic volatile organic compound (BVOC) emissions from forest ecosystems because of their contribution to tropospheric oxidation processes and secondary aerosol formation [1, 2]. It became apparent that biogenic emissions show much more variation than previously assumed. In this poster we focus on the change in BVOC emission patterns from a four year old Fagus sylvatica L. during a growth chamber experiment (PAR, temperature controlled) lasting from March to November 2008. A dynamic branch enclosure system was used in our experiments. Ozone and VOC were removed from air entering the cuvette, as ozone level was found to be a critical parameter for degradation of the compounds [3]. Samples were collected on Tenax TA-Carbotrap solid phase adsorbent tubes and analyzed by TD-GC-MS. Measurements started before budburst of the tree and finished at the end of autumn. Over the entire period 33 samples have been analyzed, while 16 compounds were detected, including 10 monoterpenes (MT), 2 oxygenated-MTs, 2 sesquiterpenes (SQT), isoprene and methyl salicylate. Sabinene showed the highest emission, in an agreement with previous studies [4, 5]. Quantifiable emission appeared 21 days after budburst, and reached the highest level at the beginning of summer. MT emissions showed a clear trend in following each other. As an illustration the trend of sabinene and limonene emission is presented. In the middle of autumn phytophaga infection was observed on the tree induced by Two-spotted mite (Tetranychus urticae). New compounds appeared as a result of infection (linalool, methyl salicylate, (E,E)-?-farnesene, unknown oxygenated-MT, unknown SQT) and became dominant over sabinene, explained by the low MT emissions at this time of the year. These observations point at the importance of further investigation of BVOC emissions (especially SQTs and oxygenated-MTs) and the need for a proper quantification system of these compounds. We would like to emphasize our acknowledgement to BELSPO (Belgian Science Policy; IMPECVOC contract # SD/TE/03A) and to FWO (Fonds voor Wetenschappelijk Onderzoek; contract # B/07659/02) for supporting the research program. [1] Ortega et al. (2008) Chemosphere (72) 365-380 [2] Duhl et al. (2008) Biogeosciences (5) 761-777 [3] Pollmann et al. (2005) Environ. Sci. Technol. (39) 9620-9629 [4] Moukhtar et al. (2005) Atmos. Environ. (39) 3535-3547 [5] Holzke et al. (2006) J. Atmos. Chem. (55) 81-102

  16. Biofilm formation of Francisella noatunensis subsp. orientalis

    USGS Publications Warehouse

    Soto, Esteban; Halliday-Wimmonds, Iona; 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 the iglC 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.

  17. Biofilm formation of Francisella noatunensis subsp. orientalis.

    PubMed

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

    2015-12-31

    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 24h where both salinity as well as temperature plays a role in the growth and biofilm formation of Fno. Mutations in the iglC 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. PMID:26507830

  18. Simultaneous detection of Clavibacter michiganensis subsp. nebraskensis and Pantoea stewartii subsp. stewartii based on microsphere immunoreaction.

    PubMed

    Zhang, Fan; Li, Jinfeng; Zou, Mingqiang; Chen, Yan; Wang, Yanfei; Qi, Xiaohua

    2013-04-01

    Clavibacter michiganensis subsp. nebraskensis (Cmn) and Pantoea stewartii subsp. stewartii (Pss) are two plant pathogens that can cause tremendous agricultural economic losses. This novel method based on microsphere immunoreaction was developed for the simultaneous detection of Cmn and Pss in maize. This multiplex method was constructed based on microsphere immunodetection with fluorescent labels such as quantum dots (QDs) and R-phycoerythrin (R-PE) for the detection of Cmn and Pss. Captured QDs and R-PE serve as signal reporters for fluorescent readout. The principle of this method is based on a sandwich immunoreaction. Cmn and Pss captured by the microspheres were detected using flow cytometry. The limit of detection of this method was 10 times lower than the enzyme-linked immunosorbent assay (ELISA), and its analysis time (1 h) was much shorter compared with ELISA (6-8 h). The method, which has been proven to be an effective approach to multiplex detection of plant bacteria (Cmn and Pss as models), not only increased the varieties but also improved the sensitivity. The microsphere immunoreaction provides a universal method for the multiplex determination of microbes because of its high sensitivity, specificity, and speed. In the future, the method will be more fully validated in vivo to detect diversiform bacteria. PMID:23169888

  19. Unraveling the growth determinism of Fagus sylvatica: a hybrid data-model approach

    NASA Astrophysics Data System (ADS)

    Guillemot, Joannès; Martin-StPaul, Nicolas; Delpierre, Nicolas; François, Christophe; Soudani, Kamel; Restoux, Gwendal; Dufrêne, Eric

    2013-04-01

    The physiological processes underlying the limitation of forest growth are still under debate. Growth has long been considered as a carbone (C) limited process (Sala et al., 2012). As a matter of facts, a recent global meta-analysis has shown good agreements between assimilated C and forest productivity (Litton et al., 2007). Consequently, a majority of the process-based productivity models considers growth as a fraction of the net primary production (NPP) (Lacointe et al., 2000; Sitch et al., 2003. However, investigations at the stand scale report conflicting results (Rocha et al., 2006, Mund et al., 2010) and are not systematically consistent with a strict C limitation of growth, thus challenging the C-centric paradigm. The mechanisms that potentially degrade the link between NPP and growth include: i) the direct effect of environmental factors on growth (Zweifel et al., 2006, Körner et al., 2003), ii) the temporal variability of the growth allocation coefficient, due either to ontogeny (Genet et al., 2009), or to the initial physiological state of the tree i.e. to the reaction to past conditions. Indeed, many dendrochronological and ecological studies have shown a correlation between growth and climatic factors of the previous years (e.g. Lebourgeois et al., 2005; Richardson et al., 2012). In this work, we used a hybrid data model approach in order to assess the determinant of Fagus sylvatica stem growth along a spatial gradient across France. Despite they could brought essential insight on tree functioning, intra-specific studies across contrasted sites are still lacking in the current debate. Standardized annual growth data series at the stand scale were calculated using circumference inventories and dendrochronological series on 17 plots of the RENECOFOR network. We used the process-based model CASTANEA, thoroughly validated in long term flux simulation across Europe (e.g. Delpierre et al. 2009), to simulate the annual NPP of the corresponding periods. We then investigated the dependency of annual growth on NPP, age and variables describing direct or indirect climatic conditions of the current (n) and the previous (n-1) year (e.g. simulated soil water content, sum of winter negative temperature, simulated growth phenology). Analyses were conducted as follows: first we used a Random Forest learning machine (RF) to select important variables and to obtain information on the form of the dependencies. We then formalized this information within the linear model framework to quantify the variability attributable to each factors and to conduct valid statistical tests. RF analysis revealed that NPP was the most important variable explaining growth, with a quasi-linear partial effect. The 2 other selected variables quantified the hydric stress of year (n) and (n-1), and impacted growth with high threshold effects. Analysis of covariance then revealed that both hydric variables significantly affect the NPP-growth relationship. The retained variables explained 29% of the growth variability. Growth of F. sylvatica is under complex control, involving C assimilation, direct effect of water shortage and lagged response to past condition. We used these results to implement in the CASTANEA model a new conceptual allocation scheme, validated at regional scale.

  20. Description and history of Syringa oblata subsp. oblata 'Frank Meyer'

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An accession of Syringa oblata subsp. oblata (PI 23031) collected in China by Frank Meyer in 1908was given the name ‘Frank Meyer’ by Father Fiala in 1988. To be established according to the International Code of Nomenclature for Cultivated Plants, a new cultivar name must be accompanied by a descrip...

  1. TARGETING MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS IN THE ENVIRONMENT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diagnosis of Johne’s disease, an enteric infection of cattle caused by Mycobacterium avium subsp. paratuberculosis (MAP), has been impeded by the lack of rapid, reliable detection methods. The goal of this study was to optimize detection of MAP in environmental samples. Experiments were conducted to...

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

    PubMed Central

    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. PMID:24714388

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

    PubMed Central

    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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  5. Cellular Interactions in Mycobacterium avium subsp. paratuberculosis Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The study of host immune responses to Mycobacterium avium subsp. paratuberculosis (MAP) is complicated by a number of factors, including the protracted nature of the disease and the stealthy nature of the pathogen. Noted as one of the more fastidious mycobacteria, infection with MAP is often chara...

  6. 40 CFR 180.41 - Crop group tables.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Almond (Prunus dulcis) Beech nut (Fagusspp.) Brazil nut (Bertholletia excelsa) Butternut (Juglans cinerea...Beechnut (Fagus grandifolia Ehrh.; F. sylvaticaL.) Brazil nut (Bertholletia excelsaHumb. & Bonpl.)...

  7. 40 CFR 180.41 - Crop group tables.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Almond (Prunus dulcis) Beech nut (Fagusspp.) Brazil nut (Bertholletia excelsa) Butternut (Juglans cinerea...Beechnut (Fagus grandifolia Ehrh.; F. sylvaticaL.) Brazil nut (Bertholletia excelsaHumb. & Bonpl.)...

  8. Alpha-glucosidase inhibitory constituents of Linaria kurdica subsp. eriocalyx.

    PubMed

    Aydo?du, Irfan; Zihnio?lu, Figen; Karayildirim, Tamer; Gülcemal, Derya; Alanku?-Cali?kan, Ozgen; Bedir, Erdal

    2010-06-01

    Three known iridoid glycosides, antirrhide (1), antirrhinoside (2), and 5-O-beta-allosylantirrhinoside (3), and two known flavone glycosides, linariin (4"'-O-acetylpectolinarin) (4) and linarin (acacetin-7-O-beta-D-rutinoside) (5) were isolated from Linaria kurdica Boiss & Hohen. subsp. eriocalyx. The structures of the isolated compounds were established from spectroscopic evidence. Compounds 1-3 showed high inhibitory potential against alpha-glucosidase. PMID:20614804

  9. PEDIOCIN PRODUCTION IN MILK BY PEDIOCOCCUS ACIDILACTICI IN CO-CULTURE WITH STREPTOCOCCUS THERMOPHILUS AND LACTOBACILLUS DELBRUECKII SUBSP. BULGARICUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The production of pediocin in milk by Pediococcus acidilactici was evaluated in co-culture with the dairy fermentation cultures Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis subsp. lactis. The cultures were tested singly or in different combinations...

  10. Comparison of pectic enzymes produced by Erwinia chrysanthemi, Erwinia carotovora subsp. carotovora, and Erwinia carotovora subsp. atroseptica.

    PubMed Central

    Ried, J L; Collmer, A

    1986-01-01

    Erwinia spp. that cause soft-rot diseases in plants produce a variety of extracellular pectic enzymes. To assess the correlation between patterns of pectic enzyme production and taxonomic classification, we compared the enzymes from representative strains. Supernatants obtained from polygalacturonate-grown cultures of nine strains of Erwinia chrysanthemi, three strains of E. carotovora subsp. carotovora, and three strains of E. carotovora subsp. atroseptica were concentrated and subjected to ultrathin-layer polyacrylamide gel isoelectric focusing. Pectate lyase, polygalacturonase, and exo-poly-alpha-D-galacturonosidase activities were visualized by staining diagnostically buffered pectate-agarose overlays with ruthenium red after incubation of the overlays with the isoelectric focusing gels. The isoelectric focusing profiles of pectate lyase and polygalacturonase were nearly identical for strains of E. carotovora subsp. carotovora and E. carotovora subsp. atroseptica, showing three pectate lyase isozymes with isoelectric points higher than 8.7 and a polygalacturonase with pI of ca. 10.2. Isoelectric focusing profiles of the E. chrysanthemi pectic enzymes were substantially different. Although there was considerable intraspecific heterogeneity, all strains produced at least four isozymes of pectate lyase, which could be divided into three groups: basic (pI, ca. 9.0 to 10.0), slightly basic (pI, ca. 7.0 to 8.5), and acidic (pI, ca. 4.0 to 5.0). Several strains of E. chrysanthemi also produced a single form of exo-poly-alpha-D-galacturonosidase (pI, ca. 8.0).(ABSTRACT TRUNCATED AT 250 WORDS) Images PMID:3752996

  11. Phosphite Protects Fagus sylvatica Seedlings towards Phytophthora plurivora via Local Toxicity, Priming and Facilitation of Pathogen Recognition

    PubMed Central

    Dalio, Ronaldo J. D.; Fleischmann, Frank; Humez, Martina; Osswald, Wolfgang

    2014-01-01

    Phytophthora plurivora causes severe damage on Fagus sylvatica and is responsible for the extensive decline of European Beech throughout Europe. Unfortunately, no effective treatment against this disease is available. Phosphite (Phi) is known to protect plants against Phytophthora species; however, its mode of action towards P. plurivora is still unknown. To discover the effect of Phi on root infection, leaves were sprayed with Phi and roots were subsequently inoculated with P. plurivora zoospores. Seedling physiology, defense responses, colonization of root tissue by the pathogen and mortality were monitored. Additionally the Phi concentration in roots was quantified. Finally, the effect of Phi on mycelial growth and zoospore formation was recorded. Phi treatment was remarkably efficient in protecting beech against P. plurivora; all Phi treated plants survived infection. Phi treated and infected seedlings showed a strong up-regulation of several defense genes in jasmonate, salicylic acid and ethylene pathways. Moreover, all physiological parameters measured were comparable to control plants. The local Phi concentration detected in roots was high enough to inhibit pathogen growth. Phi treatment alone did not harm seedling physiology or induce defense responses. The up-regulation of defense genes could be explained either by priming or by facilitation of pathogen recognition of the host. PMID:24489973

  12. Decomposition of beech (Fagus sylvatica) and pine (Pinus nigra) litter along an Alpine elevation gradient: Decay and nutrient release

    PubMed Central

    Berger, Torsten W.; Duboc, Olivier; Djukic, Ika; Tatzber, Michael; Gerzabek, Martin H.; Zehetner, Franz

    2015-01-01

    Litter decomposition is an important process for cycling of nutrients in terrestrial ecosystems. The objective of this study was to evaluate direct and indirect effects of climate on litter decomposition along an altitudinal gradient in a temperate Alpine region. Foliar litter of European beech (Fagus sylvatica) and Black pine (Pinus nigra) was incubated in litterbags during two years in the Hochschwab massif of the Northern Limestone Alps of Austria. Eight incubation sites were selected following an altitudinal/climatic transect from 1900 to 900 m asl. The average remaining mass after two years of decomposition amounted to 54% (beech) and 50% (pine). Net release of N, P, Na, Al, Fe and Mn was higher in pine than in beech litter due to high immobilization (retention) rates of beech litter. However, pine litter retained more Ca than beech litter. Altitude retarded decay (mass loss and associated C release) in beech litter during the first year only but had a longer lasting effect on decaying pine litter. Altitude comprises a suite of highly auto-correlated characteristics (climate, vegetation, litter, soil chemistry, soil microbiology, snow cover) that influence litter decomposition. Hence, decay and nutrient release of incubated litter is difficult to predict by altitude, except during the early stage of decomposition, which seemed to be controlled by climate. Reciprocal litter transplant along the elevation gradient yielded even relatively higher decay of pine litter on beech forest sites after a two-year adaptation period of the microbial community. PMID:26240437

  13. Molecular and phenotypic characterization of Vibrio aestuarianus subsp. francensis subsp. nov., a pathogen of the oyster Crassostrea gigas.

    PubMed

    Garnier, Matthieu; Labreuche, Yannick; Nicolas, Jean-Louis

    2008-10-01

    Eleven Vibrio isolates invading the hemolymph of live and moribund oysters (Crassostrea gigas) collected in the field and from a hatchery in France, were characterized by a polyphasic approach. Phylogenetic analysis of 16S rRNA, gyrB and toxR genes indicated high homogeneity between these strains and the Vibrio aestuarianus type strain (ATCC35048(T)), and confirmed previous 16S rRNA analysis. In contrast, DNA:DNA hybridization was from 61% to 100%, while phenotypic characters and virulence tests showed a large diversity between the strains. Nevertheless, several common characters allowed the isolates to be distinguished from the reference strain. On the basis of several distinct phenotypic characteristics, it is proposed to establish two subspecies within the V. aestuarianus spp. group, V. aestuarianus subsp. aestuarianus [D. Tison, R. Seidler, Vibrio aestuarianus: a new species from estuarine waters and shellfish, Int. J. Syst. Bacteriol. (1983) 699-702] and V. aestuarianus subsp. francensis for these French isolates. The characters that differentiate the new strains from V. aestuarianus subsp. aestuarianus(T) are virulence (positive for 63% of the isolates) and 12:0 fatty acid content. The colonies were smaller and uncoloured, whereas no growth occurred at 35 degrees C or on TCBS, and the strains did not utilize several substrates, including L-serine, alpha-cyclodextrin, D-mannitol, alpha-glycyl-L-aspartic acid, L-threonine and glucose-1-phosphate. PMID:18718730

  14. A UV Tolerant Mutant of Bacillus thuringiensis subsp. kurstaki Producing Melanin

    E-print Network

    Zaritsky, Arieh

    A UV Tolerant Mutant of Bacillus thuringiensis subsp. kurstaki Producing Melanin Deepak Saxena,1 thuringiensis subsp. kurstaki, producing a dark brown pigment, identified as melanin, was studied. Bt-m had that production of melanin by various microorganisms protects their susceptibility to oxidative damage caused

  15. Complete genome sequence of Mycobacterium avium subsp. paratuberculosis, isolated from human breast milk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp paratuberculosis is the etiologic agent of Johne’s disease. We report the draft genome sequences of six M. avium subsp paratuberculosis isolates obtained from diverse hosts including bison, cattle and sheep. These sequences will deepen our understanding of host association ...

  16. Development and Characterization of Monoclonal Antibodies and Aptamers Against Major Antigens of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Specific antibodies, available in unlimited quantities, have not been produced against Mycobacterium avium subsp. paratuberculosis, the bacterium that causes Johne’s disease (JD). To fill this gap in JD research, monoclonal antibodies (mAbs) against M. avium subsp. paratuberculosis were produced fr...

  17. Identification and Characterization of Iron Dependent Regulator (IdeR) of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP), is known for its dependency on mycobactin for its growth in laboratory media. Comparative genomics of M. tuberculosis, M. avium subsp. paratuberculosis and M. avium avium showed that mycobactin A, a gene encoding the first enzyme of mycobactin synth...

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

    PubMed

    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 2.10? CFU K. p. subsp. pneumoniae (n = 10) showed transient signs of infection and all of them recovered. All of those infected with 1.10? 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-? 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. PMID:21723409

  19. A Gene Specific to Mycobacterium avium subsp. paratuberculosis, But Only at the Transcription-translation Level

    Technology Transfer Automated Retrieval System (TEKTRAN)

    There is no known antibody that detects M. avium subsp paratuberculosis and does not cross react with other M. avium subspecies. In the present study, a monoclonal antibody was identified from mice immunized with a cell membrane fraction of M. avium subsp paratuberculosis strain K-10. This antibod...

  20. SALMONELLA ENTERICA SUBSP. ENTERICA IN CATTLE EGRET (BUBULCUS IBIS) CHICKS FROM CENTRAL TEXAS: PREVALENCE,

    E-print Network

    Mora, Miguel A.

    SALMONELLA ENTERICA SUBSP. ENTERICA IN CATTLE EGRET (BUBULCUS IBIS) CHICKS FROM CENTRAL TEXAS% to 95%. Seventeen Salmonella enterica subsp. enterica serotypes were isolated, of which the 4,5,12:i of Salmonella, but provided no evidence of transmission between these two species. Similar conclusions were

  1. Draft Genome Sequence of the First Human Isolate of the Ruminant Pathogen Mycoplasma capricolum subsp. capricolum

    PubMed Central

    Fischer, Anne; Heller, Martin; Jores, Joerg; Sachse, Konrad; Mourier, Tobias; Hansen, Anders Johannes

    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 isolate. PMID:26089408

  2. 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

    PubMed Central

    Fiers, Mark W. E. J.; Lu, Ashley; Armstrong, Karen F.

    2015-01-01

    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. PMID:26251497

  3. Tree Species Composition and Harvest Intensity Affect Herbivore Density and Leaf Damage on Beech, Fagus sylvatica, in Different Landscape Contexts

    PubMed Central

    Mangels, Jule; Blüthgen, Nico; Frank, Kevin; Grassein, Fabrice; Hilpert, Andrea; Mody, Karsten

    2015-01-01

    Most forests are exposed to anthropogenic management activities that affect tree species composition and natural ecosystem processes. Changes in ecosystem processes such as herbivory depend on management intensity, and on regional environmental conditions and species pools. Whereas influences of specific forest management measures have already been addressed for different herbivore taxa on a local scale, studies considering effects of different aspects of forest management across different regions are rare. We assessed the influence of tree species composition and intensity of harvesting activities on arthropod herbivores and herbivore-related damage to beech trees, Fagus sylvatica, in 48 forest plots in three regions of Germany. We found that herbivore abundance and damage to beech trees differed between regions and that – despite the regional differences - density of tree-associated arthropod taxa and herbivore damage were consistently affected by tree species composition and harvest intensity. Specifically, overall herbivore damage to beech trees increased with increasing dominance of beech trees – suggesting the action of associational resistance processes – and decreased with harvest intensity. The density of leaf chewers and mines was positively related to leaf damage, and several arthropod groups responded to beech dominance and harvest intensity. The distribution of damage patterns was consistent with a vertical shift of herbivores to higher crown layers during the season and with higher beech dominance. By linking quantitative data on arthropod herbivore abundance and herbivory with tree species composition and harvesting activity in a wide variety of beech forests, our study helps to better understand the influence of forest management on interactions between a naturally dominant deciduous forest tree and arthropod herbivores. PMID:25938417

  4. 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.

    PubMed

    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 23117(T)?=?BGSC 10A6(T)), and B. amyloliquefaciens subsp. amyloliquefaciens subsp. nov., with the type strain DSM 7(T)(?=?ATCC 23350(T)?=?Fukumoto Strain F(T)), for plant-associated and non-plant-associated representatives, respecitvely. This is in agreement with results of DDH and M-CGH tests and the MALDI-TOF MS of cellular components, all of which suggested that the ecovars represent two different subspecies. PMID:20817842

  5. Strong genetic differentiation between North American and European populations of Phytophthora alni subsp. uniformis.

    PubMed

    Aguayo, Jaime; Adams, Gerard C; Halkett, Fabien; Catal, Mursel; Husson, Claude; Nagy, Zoltán Á; Hansen, Everett M; Marçais, Benoît; Frey, Pascal

    2013-02-01

    Alder decline caused by Phytophthora alni has been one of the most important diseases of natural ecosystems in Europe during the last 20 years. The emergence of P. alni subsp. alni -the pathogen responsible for the epidemic-is linked to an interspecific hybridization event between two parental species: P. alni subsp. multiformis and P. alni subsp. uniformis. One of the parental species, P. alni subsp. uniformis, has been isolated in several European countries and, recently, in North America. The objective of this work was to assess the level of genetic diversity, the population genetic structure, and the putative reproduction mode and mating system of P. alni subsp. uniformis. Five new polymorphic microsatellite markers were used to contrast both geographical populations. The study comprised 71 isolates of P. alni subsp. uniformis collected from eight European countries and 10 locations in North America. Our results revealed strong differences between continental populations (Fst = 0.88; Rst = 0.74), with no evidence for gene flow. European isolates showed extremely low genetic diversity compared with the North American collection. Selfing appears to be the predominant mating system in both continental collections. The results suggest that the European P. alni subsp. uniformis population is most likely alien and derives from the introduction of a few individuals, whereas the North American population probably is an indigenous population. PMID:23095465

  6. Tree-Ring Stable Isotopes Reveal Twentieth-Century Increases in Water-Use Efficiency of Fagus sylvatica and Nothofagus spp. in Italian and Chilean Mountains

    PubMed Central

    Tognetti, Roberto; Lombardi, Fabio; Lasserre, Bruno; Cherubini, Paolo; Marchetti, Marco

    2014-01-01

    Changes in intrinsic water use efficiency (iWUE) were investigated in Fagus sylvatica and Nothofagus spp. over the last century. We combined dendrochronological methods with dual-isotope analysis to investigate whether atmospheric changes enhanced iWUE of Fagus and Nothofagus and tree growth (basal area increment, BAI) along latitudinal gradients in Italy and Chile. Post-maturation phases of the trees presented different patterns in ?13C, ?13C, ?18O, Ci (internal CO2 concentration), iWUE, and BAI. A continuous enhancement in isotope-derived iWUE was observed throughout the twentieth century, which was common to all sites and related to changes in Ca (ambient CO2 concentration) and secondarily to increases in temperature. In contrast to other studies, we observed a general increasing trend of BAI, with the exception of F. sylvatica in Aspromonte. Both iWUE and BAI were uncoupled with the estimated drought index, which is in agreement with the absence of enduring decline in tree growth. In general, ?13C and ?18O showed a weak relationship, suggesting the major influence of photosynthetic rate on Ci and ?13C, and the minor contribution of the regulation of stomatal conductance to iWUE. The substantial warming observed during the twentieth century did not result in a clear pattern of increased drought stress along these latitudinal transects, because of the variability in temporal trends of precipitation and in specific responses of populations. PMID:25398040

  7. The influence of O3, NO2 and SO2 on growth of Picea abies and Fagus sylvatica in the Carpathian Mountains.

    PubMed

    Muzika, R M; Guyette, R P; Zielonka, T; Liebhold, A M

    2004-07-01

    At 17 long-term pollution monitoring sites throughout the Carpathian Mountains, tree growth patterns and variation in growth rate were examined to determine relationship of tree growth to specific pollutants. Canopy dominant Picea abies and Fagus sylvatica were selected at each site. Basal area increment (BAI) values were calculated from raw ring widths and used as an estimate of tree growth. Across all sites, BAI chronologies were highly variable, therefore local conditions and forest structure accounted for considerable variation. Several significant relationships, however, implicated a role of pollutants on tree growth. Average levels (1997-1999) of NO(2) and SO(2) were inversely related to BAI means (1989-1999). Although average O(3) alone was not related to growth, the maximum O(3) value reported at the sites was negatively correlated with overall growth. A variable representing the combined effect of O(3), NO(2) and SO(2) was negatively correlated with both P. abies and F. sylvatica growth. Pollution data were used to categorize all sites into 'high' or 'low' pollution sites. Difference chronologies based on these categories indicated trends of decline in the 'high' pollution sites relative to 'low' pollution site. In the more heavily polluted sites, the BAI of Fagus sylvatica has declined approximately 50% and Picea abies has declined 20% over the past 45 years. PMID:15046841

  8. A unigene set for European beech (Fagus sylvatica L.) and its use to decipher the molecular mechanisms involved in dormancy regulation.

    PubMed

    Lesur, Isabelle; Bechade, Alison; Lalanne, Céline; Klopp, Christophe; Noirot, Céline; Leplé, Jean-Charles; Kremer, Antoine; Plomion, Christophe; Le Provost, Grégoire

    2015-09-01

    Systematic sequencing is the method of choice for generating genomic resources for molecular marker development and candidate gene identification in nonmodel species. We generated 47,357 Sanger ESTs and 2.2M Roche-454 reads from five cDNA libraries for European beech (Fagus sylvatica L.). This tree species of high ecological and economic value in Europe is among the most representative trees of deciduous broadleaf forests. The sequences generated were assembled into 21,057 contigs with MIRA software. Functional annotations were obtained for 85% of these contigs, from the proteomes of four plant species, Swissprot accessions and the Gene Ontology database. We were able to identify 28,079 in silico SNPs for future marker development. Moreover, RNAseq and qPCR approaches identified genes and gene networks regulated differentially between two critical phenological stages preceding vegetative bud burst (the quiescent and swelling buds stages). According to climatic model-based projection, some European beech populations may be endangered, particularly at the southern and eastern edges of the European distribution range, which are strongly affected by current climate change. This first genomic resource for the genus Fagus should facilitate the identification of key genes for beech adaptation and management strategies for preserving beech adaptability. PMID:25594128

  9. Identification of an outer membrane protein of Fusobacterium necrophorum subsp. necrophorum that binds with high affinity to bovine endothelial cells.

    PubMed

    Kumar, Amit; Menon, Sailesh; Nagaraja, T G; Narayanan, Sanjeev

    2015-03-23

    Fusobacterium necrophorum, a Gram-negative anaerobe, is the primary etiologic agent of liver abscesses in cattle. There are two subspecies; subsp. necrophorum and subsp. funduliforme, which differ in morphological, biochemical, molecular characteristics, and virulence. The subsp. necrophorum, which is more virulent, occurs more frequently in liver abscesses than the subsp. funduliforme. Bacterial adhesion to the host cell surface is critical to the pathogenesis of several bacterial infections, and in F. necrophorum, outer membrane proteins (OMP) have been shown to mediate adhesion to bovine endothelial cells. The objective of this study was to identify potential adhesins that are involved in adhesion of F. necrophorum subsp. necrophorum to the host cells. An OMP of 42.4 kDa, which binds with high affinity to the bovine endothelial cells and is recognized by the sera from cattle with liver abscesses, was identified. N-terminal sequencing of the protein showed 96% homology to the FomA protein of F. nucleatum. The PCR analysis showed that this fomA gene was present in several strains of subsp. necrophorum, subsp. funduliforme of bovine and subsp. funduliforme of human origin. The purified native and recombinantly expressed protein when preincubated with the endothelial cells, prevented the attachment of subsp. necrophorum significantly. In addition, the polyclonal antibody produced against the protein prevented the binding of subsp. necrophorum to bovine endothelial cells. PMID:25601800

  10. Comparative Genomic Hybridizations Reveal Genetic Regions within the Mycobacterium avium Complex That Are Divergent from Mycobacterium avium subsp. paratuberculosis Isolates†

    PubMed Central

    Paustian, Michael L.; Kapur, Vivek; Bannantine, John P.

    2005-01-01

    Mycobacterium avium subsp. paratuberculosis is genetically similar to other members of the Mycobacterium avium complex (MAC), some of which are nonpathogenic and widespread in the environment. We have utilized an M. avium subsp. paratuberculosis whole-genome microarray representing over 95% of the predicted coding sequences to examine the genetic conservation among 10 M. avium subsp. paratuberculosis isolates, two isolates each of Mycobacterium avium subsp. silvaticum and Mycobacterium avium subsp. avium, and a single isolate each of both Mycobacterium intracellulare and Mycobacterium smegmatis. Genomic DNA from each isolate was competitively hybridized with DNA from M. avium subsp. paratuberculosis K10, and open reading frames (ORFs) were classified as present, divergent, or intermediate. None of the M. avium subsp. paratuberculosis isolates had ORFs classified as divergent. The two M. avium subsp. avium isolates had 210 and 135 divergent ORFs, while the two M. avium subsp. silvaticum isolates examined had 77 and 103 divergent ORFs. Similarly, 130 divergent ORFs were identified in M. intracellulare. A set of 97 ORFs were classified as divergent or intermediate in all of the nonparatuberculosis MAC isolates tested. Many of these ORFs are clustered together on the genome in regions with relatively low average GC content compared with the entire genome and contain mobile genetic elements. One of these regions of sequence divergence contained genes homologous to a mammalian cell entry (mce) operon. Our results indicate that closely related MAC mycobacteria can be distinguished from M. avium subsp. paratuberculosis by multiple clusters of divergent ORFs. PMID:15774884

  11. Ozone-induced nitrate formation in needles and leaves of Picea abies, Fagus sylvatica and Quercus robur.

    PubMed

    Krause, G H

    1988-01-01

    Much attention has been paid to ozone as a major cause of novel forest decline in Europe. In combination with acidic mist, O(3) has been observed to increase ion leaching. Besides cations lake Mg(2+), Ca(2+), K(+), NH(4)(+), considerable amounts of nitrate were found to be leached by acidic mist from needles of Norway spruce. Controlled fumigation experiments, with 100, 300, and 600 microg O(3)m(-3) over 22 days continuously, have led to a nitrate accumulation of 94.1 +/- 14.8, 119.4 +/- 28.7 and 198.9 +/- 14.9 microg NO(3)(-1) g(-1) FW, respectively, in leaves of Quercus robur. Similar values were found in leaves of Fagus sylvatica and current and previous year needles of Picea abies. Nitrate levels of controls receiving charcoal filtered air were well below 40 microg NO(3)(-) g (-1) FW. Statistically significant elevated nitrate levels were observed after only 48 h of continuous fumigation with 600 microg O(3)m(-3), in all tree species tested, and after 144 h in the 100 microg O(3)m(-3) treatment. In another experiment, trees of Picea abies were kept in two charcoal (C) and two Purafil plus charcoal (P/C) ventilated chambers, and fumigated with O and 500 microg O(3)m(-3) in cabinets of each filter-type in order to eliminate NO(x) from chamber air. After 29 days of continuous ozone fumigation, NO(3)(-) accumulation in needles amounted to 102.0 +/- 37.7 and 137.4 +/- 40.5 microg g(-1) FW in P/C and C-filtered chambers, respectively. Nitrate contents of controls were below 30 microg NO(3)(-)g(-1) FW at the end of the experiment. No significant differences in NO(3)(-) accumulation between filter treatments were observed. Since NO(x) was reduced by more than 95% in the Purafil/charcoal versus the charcoal treatment, NO(3)(-) accumulation in needles can be attributed predominantly to the influence of ozone and not to direct NO(2) uptake of needles by the possible oxidation of NO to NO(2) in the presence of ozone. PMID:15092611

  12. Transport of soluble carbohydrates in temperate deciduous trees: beech (Fagus sylvatica) and ash (Fraxinus excelsior) in comparison

    NASA Astrophysics Data System (ADS)

    Thoms, Ronny; Köhler, Michael; Gessler, Arthur; Gleixner, Gerd

    2015-04-01

    The structure of phloem cells and the physiology of the transport of soluble carbohydrates in plants are well studied. However, the influence of different phloem un- and uploading strategies on the translocation of carbohydrates in different tree species is largely unknown. Therefore, we conducted a pulse labeling on 20 young trees of European beech (Fagus sylvatica) and European ash (Fraxinus excelsior) respectively, using the stable isotope 13C in a temperate deciduous forest in Central Germany. In one growing season each tree species was labeled in a closed transparent plastic chamber with 99% 13CO2 for 5 h. The compound specific ? 13C from carbohydrates in the different compartments leaf, branch, stem and root was measured by high-performance liquid chromatography linked with an isotope ratio mass spectrometer (HPLC-IRMS). We found that both tree species used sucrose as a transport sugar, but carbohydrates of the raffinose group (RFO) served as main transport sugar in ash trees. This indicate that beech used only the apoplastic loading strategy into the phloem cells while ash trees relied on both, apoplastic and symplastic loading, preferring the latter at the end of the growing season. Furthermore, we observed different transport velocities of labeled sugars in the two species. Here, sucrose in beech and carbohydrates of the RFO in ash were transported fastest, whereas sucrose was constantly slowest in ash trees. The label of carbohydrates was found over 60 day in the roots of both tree species, with the highest ? 13C enrichment in carbohydrates of RFO than in the other sugars. Accordingly, the mean residence time (MRT) and half life time (HLT) of 13C in different compartments were longest for carbohydrates of RFO in roots (25.6 days) and sucrose in stems (14.9 days), while the shortest MRT and HLT for sucrose appeared in beech in all compartments. Our results give evidence that RFO are preferentially transported to the root tissue as an agent against frost damage. This could have a great impact on the transport and storage behavior of sucrose in ash trees in the end of growing season. We conclude that transport mechanisms of carbohydrates in roots and stems differ. Intermediate storage of sucrose in line with different unloading strategies might be the limiting factor.

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

    PubMed

    Kaya, Duygu; Jäger, Anna K; Yalçin, Funda N; Ersöz, Tayfun

    2014-04-01

    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 plants revealed the potential effect of G. verna subsp. pontica and three benzophenone glucosides; 2,3'-dihydroxy-4-methoxy-benzophenone-6-O-beta-glucopyranoside (1), 2,4,3',4'-tetrahydroxy-benzophenone-6-O-beta-glucopyranoside (maclurin-6-O-f-glucopyranoside) (2) and 2,4,3'-trihydroxy-benzophenone-6-O-beta-glucopyranoside (3) isolated from G. verna subsp. pontica. Among the benzophenone glucosides 1 and 2 exhibited significant inhibition of MAO-A (IC50 = 31.3 +/- 4 microM and 41 +/- 4.7 microM resp.). PMID:24868869

  14. Complete genome sequence of Campylobacter fetus subsp. testudinum type strain 03-427T

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Campylobacter fetus subsp. testudinum has been isolated from reptiles and humans. This Campylobacter subspecies is genetically distinct from other C. fetus subspecies. Here we present the first whole genome sequence for this C. fetus subspecies....

  15. Comparative Genomics of Bifidobacterium animalis subsp. lactis Reveals a Strict Monophyletic Bifidobacterial Taxon

    PubMed Central

    Milani, Christian; Duranti, Sabrina; Lugli, Gabriele Andrea; Bottacini, Francesca; Strati, Francesco; Arioli, Stefania; Foroni, Elena; Turroni, Francesca; van Sinderen, Douwe

    2013-01-01

    Strains of Bifidobacterium animalis subsp. lactis are extensively exploited by the food industry as health-promoting bacteria, although the genetic variability of members belonging to this taxon has so far not received much scientific attention. In this article, we describe the complete genetic makeup of the B. animalis subsp. lactis Bl12 genome and discuss the genetic relatedness of this strain with other sequenced strains belonging to this taxon. Moreover, a detailed comparative genomic analysis of B. animalis subsp. lactis genomes was performed, which revealed a closely related and isogenic nature of all currently available B. animalis subsp. lactis strains, thus strongly suggesting a closed pan-genome structure of this bacterial group. PMID:23645200

  16. Neisseria elongata subsp elongata infective endocarditis following endurance exercise.

    PubMed

    Jenkins, Joanne May; Fife, Amanda; Baghai, Max; Dworakowski, Rafal

    2015-01-01

    A 31-year-old Argentinian woman presented with a 3-week history of fever, night sweats, myalgia and lethargy following a work trip to Uganda where she ran a marathon. Malarial screens were negative but C reactive protein, erythrocyte sedimentation rate and neutrophil count were raised and she was anaemic. A new pansystolic murmur was heard over the mitral valve and the transthoracic echocardiogram showed a large vegetation (>1?cm) with at least moderate mitral regurgitation. Blood cultures grew Neisseria elongata, subsp elongata treated initially with ceftriaxone then oral ciprofloxacin to complete 4?weeks of treatment. CT scan revealed a wedge-shaped area of low attenuation in the spleen in keeping with a splenic infarct. Seven days postadmission, the patient underwent a successful mitral valve repair. Recovery was complicated by a likely embolic infarct in the right frontal lobe, but the patient was discharged 12?days postoperative with no neurological sequelae. PMID:26655669

  17. Reassessment of Melittis melissophyllum L. subsp. melissophyllum iridoidic fraction.

    PubMed

    Venditti, A; Frezza, C; Guarcini, L; Maggi, F; Bianco, A; Serafini, M

    2016-01-01

    The analysis of the polar fraction of Melittis melissophyllum L. subsp. melissophyllum led to the identification of several iridoid glycosides: monomelittoside (1), melittoside (2), harpagide (3), acetyl-harpagide (4) and ajugoside (5). Compounds 3 and 4 are considered marker compounds for the genus and, as well as compounds 1, 2 and 5, were already evidenced in a previous study on the nominal species. It was noteworthy of the presence of allobetonicoside (6) which was never reported for this genus. The isolation of 6 is very relevant because of its allose residue on the structure. Allose has been often found in the species of the subfamily Lamioideae even if it mostly regarded flavonoids considered of chemotaxonomical relevance for some correlated genera of Lamiaceae. Same as allosyl-glycosidic flavonoids, the presence of allosyl-glycosidic iridoids may also be an additional chemosystematic evidence of botanical relationships among Lamiaceae species and genera. PMID:26131916

  18. Molecular Organization of the 25S–18S rDNA IGS of Fagus sylvatica and Quercus suber: A Comparative Analysis

    PubMed Central

    Inácio, Vera; Rocheta, Margarida; Morais-Cecílio, Leonor

    2014-01-01

    The 35S ribosomal DNA (rDNA) units, repeated in tandem at one or more chromosomal loci, are separated by an intergenic spacer (IGS) containing functional elements involved in the regulation of transcription of downstream rRNA genes. In the present work, we have compared the IGS molecular organizations in two divergent species of Fagaceae, Fagus sylvatica and Quercus suber, aiming to comprehend the evolution of the IGS sequences within the family. Self- and cross-hybridization FISH was done on representative species of the Fagaceae. The IGS length variability and the methylation level of 18 and 25S rRNA genes were assessed in representatives of three genera of this family: Fagus, Quercus and Castanea. The intergenic spacers in Beech and Cork Oak showed similar overall organizations comprising putative functional elements needed for rRNA gene activity and containing a non-transcribed spacer (NTS), a promoter region, and a 5?-external transcribed spacer. In the NTS: the sub-repeats structure in Beech is more organized than in Cork Oak, sharing some short motifs which results in the lowest sequence similarity of the entire IGS; the AT-rich region differed in both spacers by a GC-rich block inserted in Cork Oak. The 5?-ETS is the region with the higher similarity, having nonetheless different lengths. FISH with the NTS-5?-ETS revealed fainter signals in cross-hybridization in agreement with the divergence between genera. The diversity of IGS lengths revealed variants from ?2 kb in Fagus, and Quercus up to 5.3 kb in Castanea, and a lack of correlation between the number of variants and the number of rDNA loci in several species. Methylation of 25S Bam HI site was confirmed in all species and detected for the first time in the 18S of Q. suber and Q. faginea. These results provide important clues for the evolutionary trends of the rDNA 25S-18S IGS in the Fagaceae family. PMID:24893289

  19. Leaf morphology and phenology of Beech (Fagus sylvatica L.) are linked to environmental conditions depending on the altitudinal origin

    NASA Astrophysics Data System (ADS)

    Capdevielle-Vargas, Renee; Schuster, Christina; Estrella, Nicole; Menzel, Annette

    2014-05-01

    One of the principal responses of temperate climate trees to climate warming, besides migration, will be in-situ adaptation/evolution. For both, germination and growth rates can have a strong impact on survival and long-term recruitment and establishment of a species. Leaf morphology traits, together with phenology, are relevant to the study of inherent capacities of plants to adapt to an ever changing climate, especially in alpine regions, where a rapid warming has been observed in the last decades. The aim of this study was to evaluate the changes in possible adaptive traits (e.g. leaf morphology and phenology) of Beech (Fagus sylvatica L.) and to asses a decisive component of the survival strategy of this important broadly distributed Central European tree species. We collected beech seeds at six sites along two transects of a south- (900, 1000 and 1100-1400 m.a.s.l.) and a north-facing slope (800, 900 and 1100 m.a.s.l.) in 2011 (mast year) near Garmisch-Partenkirchen, Germany. All the seeds were stratified before sowing; 150 seeds were selected from each site and sowed (at the beginning of the spring) in square containers in a greenhouse under the same climatic conditions; seven phenological stages were defined following a modified beech germination key and the phenology of every seed was recorded three times a week. Harvesting took place 38/42 days after sowing and the specific leaf area (SLA), biomass, and leaf morphology (lamina length and width) were recorded for each seedling. Seeds from lower sites of the two transects presented a poorer germination rates (e.g. 30% for the south 900 m.a.s.l. site) and (75% for the north 800 m.a.s.l. site) when compared to seeds originating from higher elevations within the same transect. The highest germination percentages (98 and 85%) were observed in seeds originating from the highest elevations (e.g. 1100-1400 m.a.s.l. of the south site and 1100 m.a.s.l. of the north site, respectively). Although no significant differences in SLA were found among the altitudinal levels in any of the transects, significant differences were found in biomass among the two highest sites of the two transects. The length of the lamina differed significantly between 900 to 1100-1400 m.a.s.l. in the south facing transect, while in the north facing transect the lamina width showed significant differences between the highest and the lower sites. A higher percentage of germination of seeds originating from higher altitudinal sites may points to a developed sensitivity to environmental changes and a rapid and more favorable response. Our results suggest, contrary to what has been reported, (leaf size differentiation among altitudinal sites under natural conditions), that the altitude of origin doesn't have an overriding impact on leaf morphological responses when growing under the same conditions, indicating that leaf morphology and phenology may have an adaptive significance linked to climate.

  20. Effects of rhizopheric nitric oxide (NO) on N uptake in Fagus sylvatica seedlings depend on soil CO2 concentration, soil N availability and N source.

    PubMed

    Dong, Fang; Simon, Judy; Rienks, Michael; Lindermayr, Christian; Rennenberg, Heinz

    2015-08-01

    Rhizospheric nitric oxide (NO) and carbon dioxide (CO2) are signalling compounds known to affect physiological processes in plants. Their joint influence on tree nitrogen (N) nutrition, however, is still unknown. Therefore, this study investigated, for the first time, the combined effect of rhizospheric NO and CO2 levels on N uptake and N pools in European beech (Fagus sylvatica L.) seedlings depending on N availability. For this purpose, roots of seedlings were exposed to one of the nine combinations (i.e., low, ambient, high NO plus CO2 concentration) at either low or high N availability. Our results indicate a significant effect of rhizospheric NO and/or CO2 concentration on organic and inorganic N uptake. However, this effect depends strongly on NO and CO2 concentration, N availability and N source. Similarly, allocation of N to different N pools in the fine roots of beech seedlings also shifted with varying rhizospheric gas concentrations and N availability. PMID:26093371

  1. Growth losses in Swiss forests caused by ozone: epidemiological data analysis of stem increment of Fagus sylvatica L. and Picea abies Karst.

    PubMed

    Braun, Sabine; Schindler, Christian; Rihm, Beat

    2014-09-01

    The estimate of growth losses by ozone exposure of forest trees is a significant part in current C sequestration calculations and will also be important in future modeling. It is therefore important to know if the relationship between ozone flux and growth reduction of young trees, used to derive a Critical Level for ozone, is also valid for mature trees. Epidemiological analysis of stem increment data from Fagus sylvatica L. and Picea abies Karst. observed in Swiss forest plots was used to test this hypothesis. The results confirm the validity of the flux-response relationship at least for beech and therefore enable estimating forest growth losses by ozone on a country-wide scale. For Switzerland, these estimates amount to 19.5% growth reduction for deciduous forests, 6.6% for coniferous forests and 11.0% for all forested areas based on annual ozone stomatal uptake during the time period 1991-2011. PMID:24911370

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

    PubMed

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

    2015-12-01

    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 (p<0.05). The interaction of nisin producer L. lactis subsp. lactis GLc05 was demonstrated in situ, by demonstration of its influence in the complex microbiota naturally present in a raw goat milk cheese 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. PMID:26310130

  3. Flow Cytometric Detection of Mycobacterium avium subsp. paratuberculosis-Specific Antibodies in Experimentally Infected and Naturally Exposed Calves

    PubMed Central

    Bridger, P. S.; Bulun, H.; Fischer, M.; Akineden, Ö.; Seeger, T.; Barth, S.; Henrich, M.; Doll, K.; Bülte, M.; Menge, C.; Bauerfeind, R.

    2013-01-01

    A desirable test to diagnose infections with Mycobacterium avium subsp. paratuberculosis facilitates identification of infected cattle prior to the state of M. avium subsp. paratuberculosis shedding. This study aimed at adjusting a flow cytometry (FC)-based assay, using intact M. avium subsp. paratuberculosis bacteria as the antigen, for diagnosis of M. avium subsp. paratuberculosis infections in calves. Serum samples were collected from experimentally infected (n = 12) and naturally exposed (n = 32) calves. Samples from five calves from positive dams were analyzed to determine the dynamics of maternal antibodies. Samples from adult cattle with defined infection status served as the standard (18 M. avium subsp. paratuberculosis shedders, 22 M. avium subsp. paratuberculosis free). After preadsorption with Mycobacterium phlei, sera were incubated with M. avium subsp. paratuberculosis and M. avium subsp. avium bacterial suspensions, respectively, followed by the separate detection of bovine IgG, IgG1, IgG2, and IgM attached to the bacterial surface. M. avium subsp. paratuberculosis-specific sample/positive (S/P) ratios were compared to enzyme-linked immunosorbent assay (ELISA) S/P ratios. In adult cattle, the FC assay for IgG1 had a sensitivity of 78% at a specificity of 100%. Maternally acquired antibodies could be detected in calves up to 121 days of life. While all but two sera taken at day 100 ± 10 postnatum from naturally exposed calves tested negative, elevated S/P ratios (IgG and IgG1) became detectable from 44 and 46 weeks postinoculation onwards in two calves infected experimentally. Even with the optimized FC assay, M. avium subsp. paratuberculosis-specific antibodies can only occasionally be detected in infected calves less than 12 months of age. The failure to detect such antibodies apparently reflects the distinct immunobiology of M. avium subsp. paratuberculosis infections rather than methodological constraints. PMID:23885032

  4. Quantification of the Sensitivity of Mycobacterium avium subsp paratuberculosis and Salmonella enterica subsp enterica to Low pH and High Organic Acids using Propidium Monoazide and Quantitative PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp paratuberculosis (Map) and Salmonella enterica subsp enterica (S. enterica) are two pathogens that are a concern to food and animal safety due to their ability to withstand harsh conditions encountered in the natural environment and within the host during pathogenesis. Acid...

  5. IDENTIFICATION OF GENOMIC DIFFERENCES BETWEEN CAMPYLOBACTER JEJUNI SUBSP. JEJUNI AND CAMPYLOBACTER JEJUNI SUBSP. DOYLEI AT THE NAP LOCUS LEADS TO THE DEVELOPMENT OF A C. JEJUNI SUBSPECIATION MULTIPLEX PCR METHOD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 cli...

  6. Xanthomonas citri subsp. citri type IV Pilus is required for twitching motility, biofilm development, and adherence.

    PubMed

    Dunger, German; Guzzo, Cristiane R; Andrade, Maxuel O; Jones, Jeffrey B; Farah, Chuck S

    2014-10-01

    Bacterial type IV pili (T4P) are long, flexible surface filaments that consist of helical polymers of mostly pilin subunits. Cycles of polymerization, attachment, and depolymerization mediate several pilus-dependent bacterial behaviors, including twitching motility, surface adhesion, pathogenicity, natural transformation, escape from immune system defense mechanisms, and biofilm formation. The Xanthomonas citri subsp. citri strain 306 genome codes for a large set of genes involved in T4P biogenesis and regulation and includes several pilin homologs. We show that X. citri subsp. citri can exhibit twitching motility in a manner similar to that observed in other bacteria such as Pseudomonas aeruginosa and Xylella fastidiosa and that this motility is abolished in Xanthomonas citri subsp. citri knockout strains in the genes coding for the major pilin subunit PilAXAC3241, the ATPases PilBXAC3239 and PilTXAC2924, and the T4P biogenesis regulators PilZXAC1133 and FimXXAC2398. Microscopy analyses were performed to compare patterns of bacterial migration in the wild-type and knockout strains and we observed that the formation of mushroom-like structures in X. citri subsp. citri biofilm requires a functional T4P. Finally, infection of X. citri subsp. citri cells by the bacteriophage (?Xacm4-11 is T4P dependent. The results of this study improve our understanding of how T4P influence Xanthomonas motility, biofilm formation, and susceptibility to phage infection. PMID:25180689

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

    PubMed

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

    2011-11-01

    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. Estimation of Mycobacterium avium subsp. paratuberculosis growth parameters: strain characterization and comparison of methods.

    PubMed

    Elguezabal, Natalia; Bastida, Felix; Sevilla, Iker A; González, Nuria; Molina, Elena; Garrido, Joseba M; Juste, Ramón A

    2011-12-01

    The growth rate of Mycobacterium avium subsp. paratuberculosis was assessed by different methods in 7H9 medium supplemented with OADC (oleic acid, albumin, dextrose, catalase), Tween 80, and mycobactin J. Generation times and maximum specific growth rates were determined by wet weight, turbidometric measurement, viable count, and quantitative PCR (ParaTB-Kuanti; F57 gene) for 8 M. avium subsp. paratuberculosis strains (K10, 2E, 316F, 81, 445, 764, 22G, and OVICAP 49). Strain-to-strain differences were observed in growth curves and calculated parameters. The quantification methods gave different results for each strain at specific time points. Generation times ranged from an average of 1.4 days for viable count and qPCR to approximately 10 days for wet weight and turbidometry. The wet-weight, turbidometry, and ParaTB-Kuanti qPCR methods correlated best with each other. Generally, viability has been assessed by viable count as a reference method; however, due to M. avium subsp. paratuberculosis clumping problems and the presence of noncultivable M. avium subsp. paratuberculosis cells, we conclude that qPCR of a single-copy gene may be used reliably for rapid estimation of M. avium subsp. paratuberculosis bacterial numbers in a sample. PMID:22003015

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

    PubMed

    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 log?? 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 ?-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. PMID:22475940

  10. Infection with Mycobacterium avium subsp. paratuberculosis Results in Rapid Interleukin-1? Release and Macrophage Transepithelial Migration

    PubMed Central

    Lamont, Elise A.; O'Grady, Scott M.; Davis, William C.; Eckstein, Torsten

    2012-01-01

    Pathogen processing by the intestinal epithelium involves a dynamic innate immune response initiated by pathogen-epithelial cell cross talk. Interactions between epithelium and Mycobacterium avium subsp. paratuberculosis have not been intensively studied, and it is currently unknown how the bacterium-epithelial cell cross talk contributes to the course of infection. We hypothesized that M. avium subsp. paratuberculosis harnesses host responses to recruit macrophages to the site of infection to ensure its survival and dissemination. We investigated macrophage recruitment in response to M. avium subsp. paratuberculosis using a MAC-T bovine macrophage coculture system. We show that M. avium subsp. paratuberculosis infection led to phagosome acidification within bovine epithelial (MAC-T) cells as early as 10 min, which resulted in upregulation of interleukin-1? (IL-1?) at transcript and protein levels. Within 10 min of infection, macrophages were recruited to the apical side of MAC-T cells. Inhibition of phagosome acidification or IL-1? abrogated this response, while MCP-1/CCL-2 blocking had no effect. IL-1? processing was dependent upon Ca2+ uptake from the extracellular medium and intracellular Ca2+ oscillations, as determined by EGTA and BAPTA-AM [1,2-bis(2-aminophenoxy) ethane-N,N,N?,N?-tetraacetic acid tetrakis (acetoxymethyl ester)] treatments. Thus, M. avium subsp. paratuberculosis is an opportunist that takes advantage of extracellular Ca2+-dependent phagosome acidification and IL-1? processing in order to efficiently transverse the epithelium and enter its niche—the macrophage. PMID:22778093

  11. Persistence of Bacillus thuringiensis subsp. kurstaki in Urban Environments following Spraying?†‡

    PubMed Central

    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

  12. [The genome polymorphism of the Mycobacterium avium subsp. hominissuis strains].

    PubMed

    Starkova, D A; Mokrousov, I V; Viazovaia, A A; Zhuravlev, V Iu; Otten, T F; Vishnevski?, B I; Narvskaia, O V

    2014-01-01

    The non-tuberculosis mycobacteria Mycobacterium avium subsp. hominissuis (MAH) are able to cause human mycobacteriosis. In this work, the results of the first comprehensive study of the genome polymorphism of the clinical strains of MAH were reported using the typing scheme by 13 loci MATR-VNTR (TR292, TRX3, TR25, TR47, MATR-1, MATR-4, MATR-5, MATR-6, MATR-8, MATR-11, MATR-14, MATR-15, MATR-16) containing tandem nucleotide sites and IS1245-RFLP-typing sites. A total of 90 MAH strains isolated from patients with lung mycobacteriosis without epidemiological connection (including HIV infected) were tested in 2008-2011. The inhomogeneity of the MAH strains by 36 profiles of 13 loci MATR-VNTR was observed. The majority of the strains (68.8%) were included in the 8 MATR-VNTR clusters; most large cluster contained 37 strains with 13-bitnumerical profile 2222223145443'. The nucleotide sequence of the MATR-16 (3') locus contains the long deletion (GenBank accession no. KF479191). The MAH strains of the MATR-VNTR clusters were found to be inhomogeneous by the IS1245 marker. The MATR-VNTR-typing method by 13 loci is recommended for preliminary differentiation of domestic MAH strains with further analysis of the MATR-VNTR clusters using the IS1245-RFLP-typing method. PMID:25845136

  13. Prevalence on beef carcasses of Mycobacterium avium subsp. paratuberculosis DNA.

    PubMed

    Meadus, W J; Gill, C O; Duff, P; Badoni, M; Saucier, L

    2008-06-10

    Fifty samples were collected from each of skinned and dressed carcasses, from each of culled beef breeding cows and fed beef cattle <18 months old at two beef packing plants A and B, and from culled dairy cows at a packing plant C. The 450 samples were collected by swabbing an area of about 1000 cm2 in the anal region of each carcass. DNA extracted from each swab was tested for the IS900 and F57 sequences of the Mycobacterium avium subsp. paratuberculosis (MAP) genome by two stage, nested polymerase chain reaction (PCR) procedures. An internal amplification control (IAC) was detected in 45 or more of each group of 50 DNA preparations. IS900 and F57 were detected in some IAC-positive preparations from all and all but one of the groups of carcasses, respectively. Of the IAC-positive preparations in each group, between 6 and 54% were positive for IS900, and between 4 and 20% were positive for F57. When preparations were tested by single stage, quantitative PCR procedures, IS900 was detected in two samples but F57 was detected in none. The MAP DNA on carcasses was probably derived from small numbers of MAP from the environment that contaminated the animals' hides. PMID:18450311

  14. Bacillus thuringiensis subsp. israelensis and its dipteran-specific toxins.

    PubMed

    Ben-Dov, Eitan

    2014-04-01

    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 ?-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

  15. Esterase Activities of Fibrobacter succinogenes subsp. succinogenes S85

    PubMed Central

    McDermid, Kevin P.; MacKenzie, C. Roger; Forsberg, Cecil W.

    1990-01-01

    Cells of the anaerobic ruminal bacterium Fibrobacter succinogenes subsp. succinogenes S85 (formerly Bacteroides succinogenes) exhibit arylesterase activity. When cells were grown on cellulose, it was found that 69% of the total esterase activity was extracellular while 65% was nonsedimentable upon centrifugation of the culture supernatant at 100,000 × g. Disruption of the cells by various different methods failed to increase the esterase activity, indicating that the substrate was fully accessible to esterase enzymes in intact cells. During growth of cells with either glucose or cellulose as the sole carbon source, the increase in acetylesterase activity corresponded to an increase in cell density, suggesting constitutive production. The enzyme(s) hydrolyzed ?-naphthyl, p-nitrophenyl, and 4-methylumbelliferyl derivatives of acetic acid; xylose tetraacetate; glucose pentaacetate; acetylxylan; and a polymer composed of ferulic acid, arabinose, and xylose in molar proportions of 1:1.1:2.2 (FAX). These data demonstrate the presence of an acetylxylan esterase and a ferulic acid esterase. The cleavage of FAX also documents the presence of an ?-l-arabinofuranosidase. PMID:16348084

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

    PubMed

    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. PMID:25738552

  17. Protection from UV-B Damage of Mosquito Larvicidal Toxins from Bacillus thuringiensis subsp. israelensis Expressed in

    E-print Network

    Zaritsky, Arieh

    Protection from UV-B Damage of Mosquito Larvicidal Toxins from Bacillus thuringiensis subsp proteins of Bacillus thuringiensis subsp. israelensis from damage inflicted by UV-B, a sunlight component. israelensis as a mosquito biological control agent. Various subspecies of Bacillus thuringiensis (Bt

  18. Draft Genome Sequence of Streptococcus equi subsp. zooepidemicus Strain S31A1, Isolated from Equine Infectious Endometritis.

    PubMed

    da Piedade, Isabelle; Skive, Bolette; Christensen, Henrik; Bojesen, Anders Miki

    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. equi subsp. equi strain 4047. PMID:24009118

  19. Draft Genome Sequence of Streptococcus equi subsp. zooepidemicus Strain S31A1, Isolated from Equine Infectious Endometritis

    PubMed Central

    Skive, Bolette; Christensen, Henrik; Bojesen, Anders Miki

    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. equi subsp. equi strain 4047. PMID:24009118

  20. Refined, Circular Restriction Map of the Bacillus thuringiensis subsp. israelensis Plasmid Carrying the Mosquito Larvicidal Genes1

    E-print Network

    Zaritsky, Arieh

    Refined, Circular Restriction Map of the Bacillus thuringiensis subsp. israelensis Plasmid Carrying subsp. israelensis are located on one of its largest plasmids, nicknamed pBtoxis. Two linkage groupsHI restriction sites (Ben-Dov et al., 1996), but linking them to a single circular plasmid unambiguously

  1. Relationship between Presence of Cows with Milk Positive for Mycobacterium avium subsp. paratuberculosis-Specific Antibody by Enzyme-Linked Immunosorbent Assay and Viable M. avium subsp. paratuberculosis in Dust in Cattle Barns

    PubMed Central

    Chuchaisangrat, Ruj; Nielen, Mirjam; Koets, Ad P.

    2013-01-01

    Paratuberculosis, or Johne's disease, in cattle is caused by Mycobacterium avium subsp. paratuberculosis, which has recently been suspected to be transmitted through dust. This longitudinal study on eight commercial M. avium subsp. paratuberculosis-positive dairy farms studied the relationship between the number of cows with M. avium subsp. paratuberculosis antibody-positive milk and the presence of viable M. avium subsp. paratuberculosis in settled-dust samples, including their temporal relationship. Milk and dust samples were collected in parallel monthly for 2 years. M. avium subsp. paratuberculosis antibodies in milk were measured by enzyme-linked immunosorbent assay (ELISA) and used as a proxy for M. avium subsp. paratuberculosis shedding. Settled-dust samples were collected by using electrostatic dust collectors (EDCs) at six locations in housing for dairy cattle and young stock. The presence of viable M. avium subsp. paratuberculosis was identified by liquid culture and PCR. The results showed a positive relationship (odds ratio [OR], 1.2) between the number of cows with ELISA-positive milk and the odds of having positive EDCs in the same airspace as the adult dairy cattle. Moreover, the total number of lactating cows also showed an OR slightly above 1. This relationship remained the same for settled-dust samples collected up to 2 months before or after the time of milk sampling. The results suggest that removal of adult cows with milk positive for M. avium subsp. paratuberculosis-specific antibody by ELISA might result in a decrease in the presence of viable M. avium subsp. paratuberculosis in dust and therefore in the environment. However, this decrease is likely delayed by several weeks at least. In addition, the data support the notion that M. avium subsp. paratuberculosis exposure of young stock is reduced by separate housing. PMID:23793639

  2. 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?

    PubMed Central

    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 in this study appear to be good starter candidates. PMID:21666023

  3. Isofuranodiene is the main volatile constituent of Smyrnium perfoliatum L. subsp. perfoliatum growing in central Italy.

    PubMed

    Papa, Fabrizio; Ricciutelli, Massimo; Cianfaglione, Kevin; Maggi, Filippo

    2016-02-01

    The essential oil hydrodistilled from the aerial parts of Smyrnium perfoliatum subsp. perfoliatum growing in central Italy was analysed by GC-MS. The main peak in the gas chromatogram was given by the furanosesquiterpene curzerene which is the Cope rearrangement product of isofuranodiene formed into injector and column during the gas chromatographic run. A truthful quantification of these compounds was achieved by HPLC-DAD analysis which showed that isofuranodiene is the main volatile component (180.0 mg/g eo) of S. perfoliatum subsp. perfoliatum, while curzerene occurs in small amounts (18.1 mg/g eo). PMID:26134598

  4. [Genotypic characteristics of Mycobacterium avium subsp. hominissuis strains].

    PubMed

    Starkova, D A; Otten, T F; Mokrousov, I V; Viazovaia, A A; Vishnevski?, B I; Narvskaia, O V

    2013-09-01

    Mycobacterium avium are typical environmental, non-tuberculosis microorganisms that occasionally cause mycotuberculosis, an infectious disease in wild and domestic animals, birds, and humans. Here, we report the results of the first study on the genetic diversity of the Russian population of M. avium. A total of 85 M. avium subsp. hominissuis (MAH) clinical strains were isolated from patients (including 30 HIV-positive individuals) with mycobacteriosis in St. Petersburg, 2008-2011. The biochemical identification of the microorganisms was carried out using the PCR detection of the mobile elements IS901 and IS900, as well as of the polymorphism of restriction fragments of the hsp65 gene. The genetic diversity of the isolates was evaluated by VNTR typing based on eight variable-number tandem repeats (VNTRs) (292, X3, 25, 47, 3, 7, 10, and 32 [Thibault et al., 2007]). The MAH population studied was characterized by 15 VNTR types, including nine unique patterns and six clusters of isolates with identical eight-digit profiles. The largest clusters (22221128 and 24221128) included 45 (59.2%) and 15 (19.7%) isolates, respectively; the others contained from 2-7 strains. The strains of the cluster 2533112'8 possessed a truncated TR10 locus (allele 2'). Taking into account the absence of the epidemiological links between the patients and the fact that the infection was delivered from the environment, the high rate of clustering of MAH isolates can be explained by the low discriminatory power of the eight-locus VNTR-typing scheme (HGDI 0-0.61). PMID:25486772

  5. [Genotypic characteristics of Mycobacterium avium subsp. hominissuis strains].

    PubMed

    2013-09-01

    Mycobacterium avium are typical environmental, non-tuberculosis microorganisms that occasionally cause mycotuberculosis, an infectious disease in wild and domestic animals, birds, and humans. Here, we report the results of the first study on the genetic diversity of the Russian population of M. avium. A total of 85 M. avium subsp. hominissuis (MAH) clinical strains were isolated from patients (including 30 HIV-positive individuals) with mycobacteriosis in St. Petersburg, 2008-2011. The biochemical identification of the microorganisms was carried out using the PCR detection of the mobile elements IS901 and IS900, as well as of the polymorphism of restriction fragments of the hsp65 gene. The genetic diversity of the isolates was evaluated by VNTR typing based on eight variable-number tandem repeats (VNTRs) (292, X3, 25, 47, 3, 7, 10, and 32 [Thibault et al., 2007]). The MAH population studied was characterized by 15 VNTR types, including nine unique patterns and six clusters of isolates with identical eight-digit profiles. The largest clusters (22221128 and 24221128) included 45 (59.2%) and 15 (19.7%) isolates, respectively; the others contained from 2-7 strains. The strains of the cluster 2533112'8 possessed a truncated TR10 locus (allele 2'). Taking into account the absence of the epidemiological links between the patients and the fact that the infection was delivered from the environment, the high rate of clustering of MAH isolates can be explained by the low discriminatory power of the eight-locus VNTR-typing scheme (HGDI 0-0.61). PMID:25508903

  6. Fingerprinting of Salmonella enterica subsp. enterica serovar Enteritidis by ribotyping.

    PubMed

    Lippelt, Meike; de Isele, Theresa Sanabria; Kist, Manfred

    1997-04-01

    OBJECTIVE: To carry out an epidemiologic evaluation of Salmonella enterica subsp. enterica serovar Enteritidis outbreaks in households and small communities by means of rRNA gene restriction pattern analysis (ribotyping). METHODS: One hundred Enteritidis isolates dating from 1989 to 1994 which could be allocated epidemiologically to different sources or to small community outbreaks were investigated with ribotyping, a fingerprinting method in which bacterial DNA is hybridized with the biotin-labeled plasmid pKK 3535 containing a ribosomal RNA operon of Escherichia coli to determine the ribosomal RNA gene restriction patterns. RESULTS: Four different ribotyping patterns were found with the restriction endonuclease Smal and nine with Sphl. Ribotypes of isolates which could be allocated epidemiologically to a common source usually corresponded. Almost 60% of the Enteritidis infections had the ribotyping pattern Sphl-A. In contrast, this pattern was not found in any of the five Enteritidis strains isolated in 1989. The suspicion that Enteritidis phage type 4 infections are caused by consumption of insufficiently heated eggs is supported by the fact that the ribotyping pattern Sph1-A was found in isolates from eggs and from human specimens. CONCLUSIONS: As patterns Sphl-A and Smal-J appeared in 58% and 75% of the isolates, respectively, ribotyping cannot be used for the differentiation between various outbreaks with these two patterns. In cases where the Enteritidis strains showed less frequent patterns, ribotyping seems to be a practical tool for the identification of infection chains. In addition newly appearing ribotyping patterns can give information about the epidemiologic development of Enteritidis infection. PMID:11864109

  7. Molecular Identification of Mycobacterium avium subsp. silvaticum by Duplex High-Resolution Melt Analysis and Subspecies-Specific Real-Time PCR

    PubMed Central

    Csivincsik, Ágnes; Dán, Ádám

    2015-01-01

    Accurate identification of mycobacterial species and subspecies is essential to evaluate their significance and to perform epidemiological studies. The subspecies of Mycobacterium avium have different attributes but coincide in their zoonotic potential. Our knowledge about M. avium subsp. silvaticum is limited, since its identification is uncertain. Mycobacterium avium subsp. avium and M. avium subsp. silvaticum can be discriminated from each other based only on phenotypic characteristics, as they have almost identical genome sequences. Here we describe the development of a diagnostic method which enables the molecular identification of M. avium subsp. silvaticum and discrimination from M. avium subsp. avium based on genomic differences in a duplex high-resolution melt and M. avium subsp. silvaticum-specific mismatch real-time PCR. The developed assay was tested on reference strains and 199 field isolates, which were analyzed by phenotypic methods previously. This assay not only identified all 63 M. avium subsp. silvaticum and 138 M. avium subsp. avium strains correctly but also enabled the detection of mixed M. avium subsp. avium-M. avium subsp. silvaticum cultures. This is the first time that such a large panel of strains has been analyzed, and we also report the first isolation of M. avium subsp. silvaticum from red fox, red deer, wild boar, cattle, and badger. This assay is reliable, rapid, simple, inexpensive, and robust. It eliminates the long-existing problem of ambiguous phenotypic identification and opens up the possibility for detailed and comprehensive strain studies. PMID:25740770

  8. Molecular identification of Mycobacterium avium subsp. silvaticum by duplex high-resolution melt analysis and subspecies-specific real-time PCR.

    PubMed

    Rónai, Zsuzsanna; Csivincsik, Ágnes; Dán, Ádám

    2015-05-01

    Accurate identification of mycobacterial species and subspecies is essential to evaluate their significance and to perform epidemiological studies. The subspecies of Mycobacterium avium have different attributes but coincide in their zoonotic potential. Our knowledge about M. avium subsp. silvaticum is limited, since its identification is uncertain. Mycobacterium avium subsp. avium and M. avium subsp. silvaticum can be discriminated from each other based only on phenotypic characteristics, as they have almost identical genome sequences. Here we describe the development of a diagnostic method which enables the molecular identification of M. avium subsp. silvaticum and discrimination from M. avium subsp. avium based on genomic differences in a duplex high-resolution melt and M. avium subsp. silvaticum-specific mismatch real-time PCR. The developed assay was tested on reference strains and 199 field isolates, which were analyzed by phenotypic methods previously. This assay not only identified all 63 M. avium subsp. silvaticum and 138 M. avium subsp. avium strains correctly but also enabled the detection of mixed M. avium subsp. avium-M. avium subsp. silvaticum cultures. This is the first time that such a large panel of strains has been analyzed, and we also report the first isolation of M. avium subsp. silvaticum from red fox, red deer, wild boar, cattle, and badger. This assay is reliable, rapid, simple, inexpensive, and robust. It eliminates the long-existing problem of ambiguous phenotypic identification and opens up the possibility for detailed and comprehensive strain studies. PMID:25740770

  9. Evaluating the species- and site-specific differences in the physiological response of Picea abies, Fagus sylvatica and Larix decidua to drought

    NASA Astrophysics Data System (ADS)

    Hartl-Meier, Claudia; Rothe, Andreas; Treydte, Kerstin

    2013-04-01

    Sensitive regions like the Alps are very vulnerable to climate change. Especially warmer temperatures and a higher frequency of drought periods may imply strong effects on mountain ecosystems. In the Northern Limestone Alps, temperatures were already 1 °C higher (compared to the reference period 1941-1970) in the last two decades. Within a Bavarian-Austrian EU-project (INTERREG program) we investigated long-term growth patterns of mountain tree species and a possible growth effect caused by climate change using a dendroecological approach. In total we measured the ring widths of ~1300 living, on average 180 year old trees. The samples were taken along altitudinal gradients, ranging from 500 up to 1700 m a.s.l., in five different regions in the Northern Austrian and Bavarian Limestone Alps, covering the most prevalent coniferous (Picea abies, Abies alba, Larix decidua, Pinus sylvestris) and broad-leafed (Fagus sylvatica, Acer pseudoplatanus) mountain forest species. To get more detailed information about the physiological response to climate and especially drought events of different tree species, an additional study was conducted in the Kalkalpen Nationalpark, Austria. Stable isotopes (?13C and ?18O) of Picea abies, Fagus sylvatica and Larix decidua tree-rings (8 trees per species and site) were analysed at three different sites. The sites are located at the montane elevation level (900 m a.s.l.) on a south-facing and a north-facing slope as well on a plateau situation with deeper soils. Our main focus deals with the following questions: i) Is it possible to identify "drought events" in a region like the Alps with generally humid precipitation conditions (1400 mm/a), by analysing stable isotopes in tree rings? ii) Are there species- and/or site-specific differences in the isotopic signatures - also with respect to the trees' climate response? We will present (i) the isotopic signatures for the common period 1970-2010, (ii) their response to climate conditions, and (iii) compare them with our findings of the comprehensive tree-ring width measurements.

  10. Cyt1Aa from Bacillus thuringiensis subsp. israelensis enhances mosquitocidal activity of B. thuringiensis subsp. kurstaki HD-1 against Aedes aegypti but not Culex quinquefasciatus.

    PubMed

    Park, Hyun-Woo; Pino, Brent C; Kozervanich-Chong, Switzerlyna; Hafkenscheid, Erika A; Oliverio, Ryan M; Federici, Brian A; Bideshi, Dennis K

    2013-01-01

    The Cyt1Aa protein of Bacillus thuringiensis subsp. israelensis is known to synergize mosquitocidal proteins of B. thuringiensis and Bacillus sphaericus strains. Cyt1Aa is highly lipophilic, and after binding in vivo to the midgut microvillar membrane serves as a "receptor" for mosquitocidal Cry proteins, which subsequently form cation channels that kill mosquito larvae. Here we report that Cyt1Aa can serve a similar function for lepidopteran-specific Cry proteins of B. thuringiensis in certain mosquito larvae. Engineering Cyt1Aa into the HD-1 isolate of B. thuringiensis subsp. kurstaki enhanced toxicity against 4th instars of Aedes aegypti, but not against 4th instars of Culex quinquefasciatus. PMID:23314373

  11. Antioxidant Activity of the Essential Oils of Different Parts of Juniperus excelsa M. Bieb. subsp. excelsa and J. excelsa M. Bieb. subsp. polycarpos (K. Koch) Takhtajan (Cupressaceae)

    PubMed Central

    Emami, Sayyed Ahmad; Abedindo, Bibi Fatemeh; Hassanzadeh-Khayyat, Mohammad

    2011-01-01

    The essential oils of branchlets and fruits of Juniperus excelsa subsp. excelsa and Juniperus excelsa subsp. polycarpos were examined for their antioxidant activity. The compositions of the essential oils were studied by GC and GC-MS. To evaluation the antioxidants activity of the volatile oils, pure components and positive controls at different concentrations, thin-layer chromatography (TLC) screening methods, diphenylpicrylhydrazyl (DPPH) assay, deoxyribose degradation test and modified deoxyribose degradation test were employed. The results of the present study demonstrate some antioxidant activity for the tested essential oils obtained from various parts of both plants. It indicates that the use of these essential oils, in very low concentrations, may be useful as a natural preservative. However before any final conclusion, it is suggested that the antioxidant activity of these oils should also be evaluated by using lipid solvent system methods. PMID:24250416

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

    PubMed Central

    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. PMID:25242966

  13. 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.

    PubMed

    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)?=?LMG 27706(T)?=?DSM 27477(T)) for the BDB strains and Ralstonia pseudosolanacearum sp. nov. (type strain UQRS 461(T)?=?LMG 9673(T)?=?NCPPB 1029(T)) for the strains of R. solanacearum phylotypes I and III. PMID:24944341

  14. Characterizing the Genetic Diversity of the Clavibacter michiganensis subsp. michiganensis Population in New York.

    PubMed

    Tancos, Matthew A; Lange, Holly W; Smart, Christine D

    2015-02-01

    New York Clavibacter michiganensis subsp. michiganensis isolates, collected from disparate bacterial canker of tomato outbreaks over the past 11 years, were characterized with a multilocus sequence analysis (MLSA) scheme that differentiated the 51 isolates into 21 haplotypes with a discriminatory power of 0.944. The MLSA scheme consisted of five housekeeping genes (kdpA, sdhA, dnaA, ligA, and gyrB) and three putative pathogenicity genes (celA, tomA, and nagA). Repetitive polymerase chain reaction (PCR), with the BOX-A1R primer, confirmed the high diversity of C. michiganensis subsp. michiganensis isolates in New York by demonstrating that all six PCR patterns (A, B, 13C, 65C, 81C, and D) were present, with PCR patterns C and A being the most common. The MLSA scheme provided higher resolving power than the current repetitive-PCR approach. The plasmid profiles of New York isolates were diverse and differed from reference strain NCPPB382. PCR analysis indicated that the presence of putative pathogenicity genes varied between isolates and highlighted the ephemeral nature of pathogenicity genes in field populations of C. michiganensis subsp. michiganensis. Analysis of molecular variance between Serbian and New York C. michiganensis subsp. michiganensis isolates demonstrated that the two populations were not significantly different, with 98% genetic variation within each population and only 2% genetic variation between populations. PMID:25208240

  15. Induction of B Cell Responses Upon Experimental Infection of Neonatal Calves with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal models are useful for studying host responses to infection and aid in the development of diagnostic tools and vaccines. The current study was designed to compare the effects of different methods of experimental infection: Oral (Mycobacterium avium subsp. parauberculosis (MAP) strain K-10; Or...

  16. DETECTION OF BOVINE MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS IN CLINICAL AND ENVIRONMENTAL SAMPLES FROM AN INFECTED ANIMAL

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) causes Johne’s disease, a chronic, enteric infection that is passed from adults to calves via the fecal-oral route. Eradication of M. paratuberculosis from infected farms has been difficult and is likely due to long-ter...

  17. Cytokine Secretion in Periparturient dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne's disease, cause by Mycobacterium avium subsp. paratuberculosis (MAP), has a devastating impact on the dairy industry. Cows typically are infected as neonates, and stressors, such as parturition, may induce the transition from the subclinical to a more clinical stage of disease. The objective ...

  18. Experimental Infection of White-tailed Deer (Odocoileus virginianus) with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of paratuberculosis or Johne’s disease, a chronic enteric disease of domestic ruminants as well as some non-domestic ruminants. Paratuberculosis is characterized by a protracted subclinical phase followed by clinical signs such...

  19. Primary transcriptomes of Mycobacterium avium subsp. paratuberculosis reveal proprietary pathways in tissue and macrophages

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Mycobacterium avium subsp. paratuberculosis persistently infect intestines and mesenteric lymph nodes leading to a prolonged subclinical disease. We investigated the intracellular lifestyle of MAP in the intestines and lymph nodes to understand the MAP pathways that function to govern th...

  20. Identification of resistance to Acidovorax avenae subsp. citrulli among melon (Cucumis spp.) Plant Introductions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial fruit blotch (BFB) caused by the bacterium Acidovorax avenae subsp. citrulli (Aac) is a seed-borne disease that threatens most cucurbit crops. Although, limited resistance has been found in a small number of Plant Introductions (PI) in watermelon (Citrullus spp.), no significant activity ...

  1. Survival of Mycobacterium avium subsp. paratuberculosis in Biofilms on Livestock Watering Trough Materials

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite the ubiquitous occurrence of Mycobacterium sp. in nature and the fact that Johne’s disease has been reported worldwide, little research has been done to assess the survival of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) in agricultural environments. The goal of this stu...

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

    PubMed Central

    Rossier, Christophe; Baehler, Corinne; Schmitt, Sarah

    2014-01-01

    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. PMID:24689051

  3. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Mishmarhaemek Isolated from Bovine Feces

    PubMed Central

    Cooper, Ashley; Lambert, Dominic; Koziol, Adam G.; Seyer, Karine

    2015-01-01

    Salmonella enterica subsp. enterica serovar Mishmarhaemek is a Gram-negative, non-spore-forming, rod-shaped bacterium implicated in human clinical disease. Here, we report a 4.8-Mbp draft genome sequence of a nalidixic acid-resistant isolate of S. serovar Mishmarhaemek. PMID:26472847

  4. Induction of B Cell Responses upon Experimental Infection of Neonatal Calves with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal models are useful for studying host responses to infection and aid in the development of diagnostic tools and vaccines. The current study was designed to compare the effects of different methods of experimental infection: Oral (Mycobacterium avium subsp. parauberculosis (MAP) strain K-10; Or...

  5. Rapid Expression of Mycobacterium avium subsp. paratuberculosis Recombinant Proteins for Antigen Discovery

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne’s disease, a chronic granulomatous enteritis of ruminants and other species. Detection of infection in animals is hampered by the lack of sensitive and specific diagnostic assays. We here describe an approach that utilizes t...

  6. Genome Sequence of Lactococcus lactis subsp. cremoris Mast36, a Strain Isolated from Bovine Mastitis

    PubMed Central

    Gazzola, Simona; Fontana, Cecilia; Bassi, Daniela; Cocconcelli, Pier-Sandro; von Wright, Atte

    2015-01-01

    The genome sequence of Lactococcus lactis subsp. cremoris Mast36, isolated from bovine mastitis, is reported here. This strain was shown to be able to grow in milk and still possess genes of vegetable origin. The genome also contains a cluster of genes associated with pathogenicity. PMID:25999570

  7. Iron-sparing Response of Mycobacterium avium subsp. paratuberculosis is Strain Dependent

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Two genotypically and microbiologically distinct strains of Mycobacterium avium subsp. paratuberculosis (MAP) exist – the type I and type II strains that primarily infect sheep and cattle, respectively. Concentration of iron in the cultivation medium has been suggested as one contributin...

  8. Complete Genome Sequence of the Treponema pallidum subsp. pallidum Sea81-4 Strain

    PubMed Central

    Iverson-Cabral, Stefanie L.; King, Jordon C. K.; Molini, Barbara J.; Lukehart, Sheila A.; Centurion-Lara, Arturo

    2014-01-01

    Using the rabbit model of syphilis, the Sea81-4 strain of Treponema pallidum subsp. pallidum has been found to be more likely than other strains to invade the central nervous system (CNS). To identify possible explanations for this important phenotype at the genomic level, we sequenced the Sea81-4 strain genome. PMID:24744342

  9. From mouth to macrophage: mechanisms of innate immune subversion by Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  10. Functional Characterization of Iron Dependent Regulator (IdeR) of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study we investigated an iron dependent regulator (IdeR) of Mycobacterium avium subsp. paratuberculosis (MAP). IdeR is a transcriptional factor that plays a global iron regulatory role in Mycobacterium tuberculosis (MTB) with a 19-bp recognition sequence. IdeR recognition sites within MAP ge...

  11. GENOMIC COMPARISON OF MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS SHEEP AND CATTLE STRAINS BY MICROARRAY HYBRIDIZATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Microarray-based comparisons of three Mycobacterium avium subsp. paratuberculosis isolates, including one sheep strain and two cattle strains, identified three large genomic deletions in the sheep strain, totaling 29,208 bp and involving 24 open reading frames. These deletions may help explain some ...

  12. Molecular markers to determine ecological fate of Bacillus thuringiensis subsp. kurstaki

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus thuringiensis (“Bt”) is a ubiquitous soil bacterium with entomopathogenic properties. One strain, Bt subsp. kurstaki (“Btk”), is highly toxic to lepidopteran larvae and used in many commercial products for biological pest control. We designed a set of DNA markers that successfully identifi...

  13. Development of loop-mediated isothermal amplification for detection of Leifsonia xyli subsp. xyli in sugarcane

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ratoon stunt, caused by the xylem-limited coryneform bacterium Leifsonia xyli subsp. xyli (Lxx), is prevalent in most sugarcane-producing countries. Because the disease does not cause characteristic external symptoms, a laboratory-based technique is needed for accurate diagnosis. We developed a diag...

  14. VERTEBRATE TOXICOLOGY OF THE SOLUBILIZED PROTEINS OF BACILLUS THURINGIENSIS SUBSP. ISRAELENSIS

    EPA Science Inventory

    This review summarizes the studies done with the mammalian toxic Bacillus thuringiensis subsp. israelensis (Bti) 28 kDa cytA protein. The data is relevant to hazard identification studies with bacterial pesticides. The data shows that cytA produces lethal physiological changes in...

  15. Envelope protein complexes of Mycobacterium avium subsp. paratuberculosis and their antigenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease, a chronic enteric disease of ruminant animals. In the present study, blue native PAGE electrophoresis and 2D SDS-PAGE were used to separate MAP envelope protein complexes, followed by mass spectrometry (MS) ...

  16. Osteopontin Expression in Periparturient Dairy Cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne’s disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is estimated to infect more than 22% of US dairy herds. Periods of immunosuppression, typically seen at parturition, may contribute to the transition from the subclinical, or asymptomatic, to the clinical stage of inf...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

    PubMed Central

    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

  19. Immunologic responses to Mycobacterium avium subsp. paratuberculois protein cocktail vaccines in a mouse model

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne’s disease is a chronic granulomatous enteritis characterized by severe diarrhea, wasting and a decline in milk production caused by the bacterium Mycobacterium avium subsp. paratuberculois (MAP). The vaccine currently on the market has some limitations including a severe injection site reactio...

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

    EPA Science Inventory

    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...

  1. Survival of Mycobacterium avium subsp. paratuberculosis in Biofilms on Livestock Watering Trough Materials

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) is the causative agent of Johne’s disease, a chronic enteric infection that affects ruminants. Despite the ubiquitous occurrence of Mycobacterium sp. in nature and the fact that Johne’s disease has been reported worldwide, little rese...

  2. Top rot form of red stripe caused by Acidovirax avenae subsp. avenae in Louisiana sugarcane

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Red stripe of sugarcane caused by Acidovirax avenae subsp. avenae consists of two forms – leaf stripe and top rot. Symptoms of red stripe in Louisiana over the past 25 years have been limited to the leaf stripe form which causes no apparent yield loss. During 2010, the more severe top rot form was...

  3. Comparison of fecal DNA extraction kits for the detection of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fecal culture is considered the gold standard for the diagnostics of paratuberculosis, however, PCR for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in fecal material is widely used today, having demonstrated great sensitivity and specificity. To insure the most efficient and r...

  4. Comparison of fecal DNA extraction kits for the detection of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fecal culture is considered the gold standard for the diagnosis of paratuberculosis, however, PCR for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in fecal material is widely used today, having demonstrated great sensitivity and specificity. To insure the most efficient and rep...

  5. Parturition and Mycobacterium avium subsp. paratuberculosis: A Potential Interface for the Pathogenesis of Johne's Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne’s disease (JD), is estimated to infect more than 22% of US dairy herds and cost the industry $250 million annually. One major period of stress for dairy cows is the periparturient period, and field observations suggest ...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

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

    PubMed Central

    Oliveira, Letícia C.; Saraiva, Tessália D. L.; Soares, Siomar C.; Ramos, Rommel T. J.; Sá, Pablo H. C. G.; Carneiro, Adriana R.; Miranda, Fábio; Freire, Matheus; Renan, Wendel; Júnior, Alberto F. O.; Santos, Anderson R.; Pinto, Anne C.; Souza, Bianca M.; Castro, Camila P.; Diniz, Carlos A. A.; Rocha, Clarissa S.; Mariano, Diego C. B.; de Aguiar, Edgar L.; Folador, Edson L.; Barbosa, Eudes G. V.; Aburjaile, Flavia F.; Gonçalves, Lucas A.; Guimarães, Luís C.; Azevedo, Marcela; Agresti, Pamela C. M.; Silva, Renata F.; Tiwari, Sandeep; Almeida, Sintia S.; Hassan, Syed S.; Pereira, Vanessa B.; Abreu, Vinicius A. C.; Pereira, Ulisses P.; Dorella, Fernanda A.; Carvalho, Alex F.; Pereira, Felipe L.; Leal, Carlos A. G.; Figueiredo, Henrique C. P.; Silva, Artur; Miyoshi, Anderson

    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. PMID:25278529

  8. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Mishmarhaemek Isolated from Bovine Feces.

    PubMed

    Cooper, Ashley; Lambert, Dominic; Koziol, Adam G; Seyer, Karine; Carrillo, Catherine D

    2015-01-01

    Salmonella enterica subsp. enterica serovar Mishmarhaemek is a Gram-negative, non-spore-forming, rod-shaped bacterium implicated in human clinical disease. Here, we report a 4.8-Mbp draft genome sequence of a nalidixic acid-resistant isolate of S. serovar Mishmarhaemek. PMID:26472847

  9. Development and Application of a Multilocus Sequence Typing Scheme for Streptococcus gallolyticus subsp. gallolyticus

    PubMed Central

    Dumke, J.; Hinse, D.; Vollmer, T.; Knabbe, C.

    2014-01-01

    Streptococcus gallolyticus subsp. gallolyticus (formerly known as S. bovis biotype I) is a commensal of the gastrointestinal tract in animals and in up to 15% of healthy humans. Furthermore, it is a facultative pathogen that can cause infectious endocarditis, mastitis, and septicemia. The number of infections is increasing, but the transmission routes and zoonotic potential remain unknown. To assess the zoonotic potential and characterize the epidemiological structure of S. gallolyticus subsp. gallolyticus, we established a multilocus sequence typing (MLST) scheme. We amplified and sequenced internal fragments of seven housekeeping genes. The resulting sequences were analyzed with BioNumerics software 6.6 by using the unweighted-pair group method using average linkages algorithm. A total of 101 S. gallolyticus subsp. gallolyticus strains isolated from animals, humans, and environmental samples were analyzed and divided into 50 sequence types. Our first results highlight the importance of this MLST scheme for investigating the epidemiology, transmission patterns, and infection chains of S. gallolyticus subsp. gallolyticus. PMID:24789199

  10. Effect of Watering Trough Chlorination on Persistence of Mycobacterium avium subsp. Paratuberculosi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The continued global increase in the number of cases of Johne’s disease suggests that more information is needed to understand the mechanisms by which the causative agent Mycobacterium avium subsp. paratuberculosis (MAP) is spread among livestock on the farm site. Livestock watering troughs are freq...

  11. New Tricks from an Old Cow: Infective Endocarditis Caused by Streptococcus dysgalactiae subsp. dysgalactiae

    PubMed Central

    Jordal, Stina; Glambek, Marte; Oppegaard, Oddvar

    2014-01-01

    We present a case of infective endocarditis caused by Streptococcus dysgalactiae subsp. dysgalactiae, a major cause of bovine mastitis and previously thought to be an animal-restricted pathogen. The patient reported no direct contact with animals, and the clinical course was severe and complicated. PMID:25472489

  12. Evaluation of Mycobacterium avium subsp. paratuberculosis Transport in Saturated Porous Media

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of Johne’s disease, a chronic enteric infection that affects ruminants. Eradication of Map from infected farms has been difficult and is likely due to long-term survival of the organism in the environment. The application of Ma...

  13. Immunologic Responses to Mycobacterium avium subsp. paratuberculosis in Neonatal Calves After Oral or Intraperitoneal Experimental Infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infection models are useful for studying host responses to infection to aid in the development of diagnostic tools and vaccines. The majority of experimental models for ruminants have utilized an oral inoculation of live Mycobacterium avium subsp. paratuberculosis (MAP) in order to establish infecti...

  14. Genome Sequence of the Persistent Salmonella enterica subsp. enterica Serotype Senftenberg Strain SS209

    PubMed Central

    Boumart, Zineb; Virlogeux-Payant, Isabelle; Loux, Valentin; Chiapello, Hélène; Gendrault, Annie; Gibrat, Jean-François; Chemaly, Marianne; Velge, Philippe

    2012-01-01

    Salmonella enterica subsp. enterica serotype Senftenberg is an emerging serotype in poultry production which has been found to persist in animals and the farm environment. We report the genome sequence and annotation of the SS209 strain of S. Senftenberg, isolated from a hatchery, which was identified as persistent in broiler chickens. PMID:22493197

  15. Genome Sequence of the Invasive Salmonella enterica subsp. enterica Serotype Enteritidis Strain LA5

    PubMed Central

    Rossignol, Aurore; Loux, Valentin; Chiapello, Hélène; Gendrault, Annie; Gibrat, Jean-François; Velge, Philippe; Virlogeux-Payant, Isabelle

    2012-01-01

    Salmonella enterica subsp. enterica serotype Enteritidis is one of the major causes of gastroenteritis in humans due to consumption of poultry derivatives. Here we report the whole-genome sequence and annotation, including the virulence plasmid, of S. Enteritidis LA5, which is a chicken isolate used by numerous laboratories in virulence studies. PMID:22493198

  16. Draft Genome Sequence of the Putrescine-Producing Strain Lactococcus lactis subsp. lactis 1AA59

    PubMed Central

    Ladero, Victor; del Rio, Beatriz; Linares, Daniel M.; Fernandez, María; Mayo, Baltasar; Martín, M. Cruz

    2015-01-01

    We report here the 2,576,542-bp genome annotated draft assembly sequence of Lactococcus lactis subsp. lactis 1AA59. This strain—isolated from a traditional cheese—produces putrescine, one of the most frequently biogenic amines found in dairy products. PMID:26089428

  17. Production and Characterization of Monoclonal Antibodies Against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Mycobacterium avium subsp. paratuberculosis 35-kDa major membrane protein (MMP) encoded by MAP2121c has been shown to play a role in invasion of epithelial cells and is an important membrane antigen recognized by cattle with Johne’s disease. In this study, purified recombinant MMP was used to p...

  18. Antigenic Profiles of Recombinant Proteins from Mycobacterium avium subsp paratuberculosis in Sheep with Johne's Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Methods to improve the ELISA test to detect Mycobacterium avium subsp paratuberculosis have been explored over several years. Previously, selected recombinant proteins of M. avium subspecies paratuberculosis were found to be immunogenic in cattle with Johne’s disease. In the present study, antibo...

  19. Identification of Resistance to Acidovorax avenae subsp. citrulli among Melon (Cucumis spp.) Plant Introductions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial fruit blotch caused by the bacterium Acidovorax avenae subsp. citrulli (Aac) is a seed-borne disease that threatens most cucurbit crops. Although limited resistance has been found in a small number of Plant Introductions (PI) in watermelon (Citrullus spp.), no significant activity in scre...

  20. Whole-genome sequencing of Salmonella enterica subsp. enterica serovar Cubana strains isolated from agricultural sources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report draft genomes of Salmonella enterica subsp. enterica Serovar Cubana strain CVM42234 isolated from chick feed in 2012 and Salmonella Cubana strain 76814 isolated from swine in 2004. The genome sizes are 4,975,046 and 4,936,251 base pairs, respectively....

  1. An improved assay for detection of Acidovorax avenae subsp. citrulli in watermelon and melon seed

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acidovorax avenae subsp. citrulli (Aac), the causal agent of a watermelon seedling blight and fruit blotch (WFB), has emerged as a serious seedborne pathogen of watermelon, melons, pumpkin, and citron. Although attempts have been made to develop a simple routine laboratory seed assay to detect the...

  2. Immunlogic responses to Mycobacterium avium subsp. paratuberculosis protein cocktail vaccines in a mouse model

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne’s disease is a chronic granulomatous enteritis characterized by severe diarrhea, wasting, and a decline in milk production caused by the bacterium Mycobacterium avium subsp. paratuberculosis (MAP). The vaccine currently on the market has some limitations including a severe injection site react...

  3. GEOGRAPHIC DISTRIBUTION OF COMMON AND DWARF BUNT RESISTANCE IN LANDRACES OF TRITICUM AESTIVUM SUBSP. AESTIVUM

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Landrace accessions of cultivated bread wheat (Triticum aestivum L. subsp. aestivum) (TAA) from the USDA-ARS National Small Grains Collection (NSGC) have been tested systematically for the past 25 years for disease resistance. We analyzed the resistance of 10759 TAA accessions to common bunt (CB), c...

  4. Exploring the strain-specific attachment of Leuconostoc gelidum subsp. gasicomitatum on food contact surfaces.

    PubMed

    Pothakos, Vasileios; Aulia, Yosi Ayu; Van der Linden, Inge; Uyttendaele, Mieke; Devlieghere, Frank

    2015-04-16

    The psychrotrophic lactic acid bacterium (LAB) Leuconostoc gelidum subsp. gasicomitatum has emerged as one of the most prevalent specific spoilage organisms (SSOs) of packaged, cold-stored food products in Northern Europe. The whole genome sequencing of the type strain L. gelidum subsp. gasicomitatum LMG 18811(T) revealed genes encoding for proteins related to adhesion. In the present study the attachment of six food and environmental isolates was monitored on stainless steel (SS) and glass surfaces incubated (7 °C for 5-9 days) in two food simulating substrates (i.e. sweet bell pepper juice and boiled eggs in brine). The selection encompassed unique genotypes, isolated from different food products or sampling sites as well as slime-forming biotypes. The evaluation of the attached cells was performed with the bead vortexing method and a viability staining assay coupled with epifluorescence microscopy. On SS surfaces the slime-formers showed the lowest attachment (3.3-4.5 logCFU/cm(2)), while strain L. gelidum subsp. gasicomitatum ab2, which was isolated from an acetic acid bath in a vegetable salad company, reached significantly higher populations of attached cells exceeding 7 logCFU/cm(2). Strain ab2 formed dense cell aggregations on SS after 9 days of incubation in sweet bell pepper juice. The attachment ability of L. gelidum subsp. gasicomitatum on surfaces documented in the present study extends our knowledge and understanding of the spoilage potential and intra-subspecies diversity of this microbe. PMID:25625910

  5. High-Quality Draft Genome Sequence of Francisella tularensis subsp. holarctica Strain OR96-0246.

    PubMed

    Atkins, L M; Holder, M E; Ajami, N J; Metcalf, G A; Weissenberger, G M; Wang, M; Vee, V; Han, Y; Muzny, D M; Gibbs, R A; Petrosino, J F

    2015-01-01

    The bacterial pathogen Francisella tularensis was recently renewed as a tier-one select agent. F. tularensis subsp. tularensis (type A) and holarctica (type B) are of clinical relevance. Here, we report the complete genome of a virulent F. tularensis type B strain and describe its usefulness in comparative genomics. PMID:26272574

  6. High-Quality Draft Genome Sequence of Francisella tularensis subsp. holarctica Strain OR96-0246

    PubMed Central

    Atkins, L. M.; Holder, M. E.; Ajami, N. J.; Metcalf, G. A.; Weissenberger, G. M.; Wang, M.; Vee, V.; Han, Y.; Muzny, D. M.; Gibbs, R. A.

    2015-01-01

    The bacterial pathogen Francisella tularensis was recently renewed as a tier-one select agent. F. tularensis subsp. tularensis (type A) and holarctica (type B) are of clinical relevance. Here, we report the complete genome of a virulent F. tularensis type B strain and describe its usefulness in comparative genomics. PMID:26272574

  7. NOVEL DETECTION METHOD FOR MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS USING SURFACE-ENHANCED RAMAN SCATTERING BASED IMMUNOASSAYS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent of Johne's disease in cattle, imparts a loss of over $200 million to the U.S. dairy industry each year. One of the major obstacles in controlling the spread of this highly prevalent disease is the facile detection of the bacteri...

  8. Conditioned food aversion for control of poisoning by Ipomoea carnea subsp. fistulosa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conditioned food aversion is a technique that can be used to train livestock to avoid ingestion of poisonous plants. This study tested the efficacy and durability of conditioned food aversion to eliminate goat’s consumption of Ipomoea carnea subsp. fistulosa. We used 14 young Moxotó goats, which wer...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 comm...

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

    PubMed

    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. PMID:25835027

  11. INTERACTIONS BETWEEN BACILLUS THURINGIENSIS SUBSP. ISRAELENSIS AND FATHEAD MINNOWS, PIMEPHALES PROMELAS RAFINESQUE, UNDER LABORATORY CONDITIONS

    EPA Science Inventory

    Interactions between Bacillus thuringiensis subsp. israelensis and fathead minnows, Pimephales promelas, were studied in laboratory exposures to two commercial formulations, Vectobac-G and Mosquito Attack. ortality among fatheads exposed to 2.0 x 10 6 to 6.5 x 10 6 CFU/ml with bo...

  12. EFFECT OF REMOVAL OF THE CYTOLYTIC FACTOR OF 'BACILLUS THURINGIENSIS' SUBSP. 'ISRAELENSIS' ON MOSQUITO TOXICITY

    EPA Science Inventory

    Solubilized crystal protein of Bacillus thuringiensis subsp. israelensis was fractionated by affinity chromotography using a monoclonal antibody directed against the crystal's 28 kDa peptide. The 28 kDa peptide ws found to be relatively nontoxic to mosquito larvae although it doe...

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

    PubMed

    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. PMID:25816922

  14. Age-related changes in protein metabolism of beech (Fagus sylvatica L.) seeds during alleviation of dormancy and in the early stage of germination.

    PubMed

    Ratajczak, Ewelina; Kalemba, Ewa M; Pukacka, Stanislawa

    2015-09-01

    The long-term storage of seeds generally reduces their viability and vigour. The aim of this work was to evaluate the effect of long-term storage on beech (Fagus sylvatica L.) seeds at optimal conditions, over 9 years, on the total and soluble protein levels and activity of proteolytic enzymes, including endopeptidases, carboxypeptidases and aminopeptidases, as well as free amino acid levels and protein synthesis, in dry seeds, after imbibition and during cold stratification leading to dormancy release and germination. The same analyses were conducted in parallel on seeds gathered from the same tree in the running growing season and stored under the same conditions for only 3 months. The results showed that germination capacity decreased from 100% in freshly harvested seeds to 75% in seeds stored for 9 years. The levels of total and soluble proteins were highest in freshly harvested seeds and decreased significantly during storage, these proportions were retained during cold stratification and germination of seeds. Significant differences between freshly harvested and stored seeds were observed in the activities of proteolytic enzymes, including endopeptidases, aminopeptidases and carboxypeptidases, and in the levels of free amino acids. The neosynthesis of proteins during dormancy release and in the early stage of seed germination was significantly weaker in stored seeds. These results confirm the importance of protein metabolism for seed viability and the consequences of its reduction during seed ageing. PMID:26071872

  15. Ionic charge, radius, and potential control root/soil concentration ratios of fifty cationic elements in the organic horizon of a beech (Fagus sylvatica) forest podzol.

    PubMed

    Tyler, Germund

    2004-08-15

    The root/organic soil concentration ratio; R/S) of 50 cationic mineral elements was related to their ionic properties, including ionic radius (r), ionic charge (z), and ionic potential (z/r or z2/r). The materials studied were ectomycorrhizal beech (Fagus sylvatica L.) roots and their almost purely organic soil substrate, the O-horizon (mor; raw humus) of a Podzol in South Sweden, developed in a site which has been untouched by forestry or other mechanical disturbance since at least 50 years and located in an area with no local sources of pollution. Elements determined by ICP-AES were aluminium, barium, calcium, iron, potassium, magnesium, manganese, sodium and strontium. Determined by ICP-MS were silver, beryllium, bismuth, cadmium, cerium, cobalt, chromium, caesium, copper, dysprosium, erbium, europium, gallium, gadolinium, hafnium, mercury, holmium, indium, lanthanum, lithium, lutetium, niobium, neodymium, nickel, lead, praseodymium, rubidium, scandium, samarium, tin, terbium, thorium, titanium, thallium, thulium, uranium, vanadium, yttrium, ytterbium, zinc and zirconium. The R/S ratios were most clearly related to the ionic potential of the cationic elements studied, which accounted for approximately 60% of the variability in R/S among elements. The ionic charge of an element was more important than the ionic radius. Elements with high ionic charge had low R/S ratios and vice versa. No clear differences in R/S between essential and non-essential plant nutrients were observed, especially when ions of similar charge were compared. PMID:15262169

  16. Influence of the moisture content on the composition of the liquid smoke produced in the pyrolysis process of Fagus sylvatica L. wood.

    PubMed

    Guillén, M D; Ibargoitia, M L

    1999-10-01

    The pyrolysis of several samples of sawdust of Fagus sylvatica L. wood with different moisture contents was carried out, keeping all other smoke generation parameters constant. However, parameters such as smoke production length and maximum temperature reached were affected by the moisture content of the sample and varied in the different pyrolytic runs. The acidity and the composition of the liquid smokes obtained were determined, this latter by means of gas chromatography/mass spectrometry and gas chromatography with flame ionization detection. The acidity and composition of the liquid smoke produced were affected not only by the moisture content of the sawdust sample but also by the smoke generation length and by the temperature of the process. The highest yields in components were produced from samples with low moisture content that underwent a short pyrolytic process. Some compounds, with important properties from an organoleptic and preservative point of view, were not generated from samples with high moisture content. Equations that closely relate yield of the total components or of groups of components or of individual components with parameters such as moisture content, length of the process, and temperature were obtained; these equations predict yield data of liquid smoke components with a satisfactory degree of approximation. PMID:10552778

  17. Influence of type of culture medium on characterization of Mycobacterium avium subsp. paratuberculosis subtypes.

    PubMed

    Cernicchiaro, N; Wells, S J; Janagama, H; Sreevatsan, S

    2008-01-01

    We evaluated the effects of culture and/or enrichment methods on the selection of Mycobacterium avium subsp. paratuberculosis subtypes. M. avium subsp. paratuberculosis isolates from bovine fecal samples processed using a centrifugation protocol were investigated in both liquid (MGIT ParaTb tubes) and solid (Herrold's egg yolk medium) culture media. For this evaluation, M. avium subsp. paratuberculosis subtyping was based on the sequence variation in two of the most discriminatory short sequence repeat loci, i.e., mononucleotide G and trinucleotide GGT, in isolates from liquid and solid cultures. This study identified the existence of one major predominating fingerprint (>13G-5GGT) in bovine fecal samples, regardless of the type of medium used for isolation. Matched-pair analysis of subtypes showed that 69% of samples presented unique subtypes in the two culture media used, while 31% shared the same G-GGT allele. Furthermore, the liquid culture method appeared to select for a more genotypically diverse subtype population than the solid culture method. The variety of subtypes observed between liquid and solid cultures obtained from the same fecal samples suggests that the culture method could provide a "microbiological" bias and lead to a discrepancy in the growth of M. avium subsp. paratuberculosis strains. In conclusion, this study identified that these two types of culture media determined differential growth of M. avium subsp. paratuberculosis strains and that this should be considered in evaluating detection capabilities of diagnostic tests or interpreting data from molecular epidemiological studies performed using conventional solid fecal culture or automated liquid medium culture methods. PMID:17959762

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

    PubMed Central

    2011-01-01

    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. PMID:21824414

  19. Discovery of Stable and Variable Differences in the Mycobacterium avium subsp. paratuberculosis Type I, II, and III Genomes by Pan-Genome Microarray Analysis? †

    PubMed Central

    Castellanos, Elena; Aranaz, Alicia; Gould, Katherine A.; Linedale, Richard; Stevenson, Karen; Alvarez, Julio; Dominguez, Lucas; de Juan, Lucia; Hinds, Jason; Bull, Tim J.

    2009-01-01

    Mycobacterium avium subsp. paratuberculosis is an important animal pathogen widely disseminated in the environment that has also been associated with Crohn's disease in humans. Three M. avium subsp. paratuberculosis genomotypes are recognized, but genomic differences have not been fully described. To further investigate these potential differences, a 60-mer oligonucleotide microarray (designated the MAPAC array), based on the combined genomes of M. avium subsp. paratuberculosis (strain K-10) and Mycobacterium avium subsp. hominissuis (strain 104), was designed and validated. By use of a test panel of defined M. avium subsp. paratuberculosis strains, the MAPAC array was able to identify a set of large sequence polymorphisms (LSPs) diagnostic for each of the three major M. avium subsp. paratuberculosis types. M. avium subsp. paratuberculosis type II strains contained a smaller genomic complement than M. avium subsp. paratuberculosis type I and M. avium subsp. paratuberculosis type III genomotypes, which included a set of genomic regions also found in M. avium subsp. hominissuis 104. Specific PCRs for genes within LSPs that differentiated M. avium subsp. paratuberculosis types were devised and shown to accurately screen a panel (n = 78) of M. avium subsp. paratuberculosis strains. Analysis of insertion/deletion region INDEL12 showed deletion events causing a reduction in the complement of mycobacterial cell entry genes in M. avium subsp. paratuberculosis type II strains and significantly altering the coding of a major immunologic protein (MPT64) associated with persistence and granuloma formation. Analysis of MAPAC data also identified signal variations in several genomic regions, termed variable genomic islands (vGIs), suggestive of transient duplication/deletion events. vGIs contained significantly low GC% and were immediately flanked by insertion sequences, integrases, or short inverted repeat sequences. Quantitative PCR demonstrated that variation in vGI signals could be associated with colony growth rate and morphology. PMID:19047395

  20. Osteopontin Immunoreactivity in the Ileum and Ileoceccal Lymph Node of Dairy Cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Osteopontin (Opn), a highly acidic glycoprotein, promotes cellular adhesion and recruitment and has been shown to be upregulated in the granulomas of mycobacterial infections. Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is associated with granulomatous enteritis. ...

  1. Osteopontin: A Novel Cytokine Involved in the Regulation of Mycobacterium avium subsp. paratuberculosis Infection in Periparturient Dairy Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Osteopontin (Opn), an important mediator of the cell-mediated immune response, enhances the host immune response against mycobacterial infections. Infections caused by the intracellular bacterium, Mycobacterium avium subsp. paratuberculosis (MAP), have a devastating impact on the dairy industry. ...

  2. HAS MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS ACQUIRED VIRULENCE FACTORS TO COMPENSATE FOR THE ABSENCE OF PE/PGRS/PPE GENES?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial genomes are constantly evolving to increase adaptability to their changing surroundings and generate evolutionary novelty. Comparison of the five sequenced mycobacterial genomes reveals that while Mycobacterium lepare has evolved by retaining a minimal gene set, Mycobacterium avium subsp. ...

  3. Contrasting results of culture-dependent and molecular analyses of Mycobacterium avium subsp. paratuberculosis from wood bison.

    PubMed

    Forde, Taya; De Buck, Jeroen; Elkin, Brett; Kutz, Susan; van der Meer, Frank; Orsel, Karin

    2013-07-01

    Reduced to near extinction in the late 1800s, a number of wood bison populations (Bison bison athabascae) have been re-established through reintroduction initiatives. Although an invaluable tool for conservation, translocation of animals can spread infectious agents to new areas or expose animals to pathogens in their new environment. Mycobacterium avium subsp. paratuberculosis, a bacterium that causes chronic enteritis in ruminants, is among the pathogens of potential concern for wood bison management and conservation. In order to inform translocation decisions, our objectives were to determine the M. avium subsp. paratuberculosis infection status of wood bison herds in Canada and to culture and genetically characterize the infective strain(s). We tested fecal samples from bison (n = 267) in nine herds using direct PCR for three M. avium subsp. paratuberculosis-specific genetic targets with different copy numbers within the M. avium subsp. paratuberculosis genome. Restriction enzyme analysis (REA) and sequencing of IS1311 were performed on seven samples from five different herds. We also evaluated a panel of different culture conditions for their ability to support M. avium subsp. paratuberculosis growth from feces and tissues of direct-PCR-positive animals. Eighty-one fecal samples (30%) tested positive using direct IS900 PCR, with positive samples from all nine herds; of these, 75% and 21% were also positive using ISMAP02 and F57, respectively. None of the culture conditions supported the growth of M. avium subsp. paratuberculosis from PCR-positive samples. IS1311 REA and sequencing indicate that at least two different M. avium subsp. paratuberculosis strain types exist in Canadian wood bison. The presence of different M. avium subsp. paratuberculosis strains among wood bison herds should be considered in the planning of translocations. PMID:23686265

  4. Contrasting Results of Culture-Dependent and Molecular Analyses of Mycobacterium avium subsp. paratuberculosis from Wood Bison

    PubMed Central

    De Buck, Jeroen; Elkin, Brett; Kutz, Susan; van der Meer, Frank; Orsel, Karin

    2013-01-01

    Reduced to near extinction in the late 1800s, a number of wood bison populations (Bison bison athabascae) have been re-established through reintroduction initiatives. Although an invaluable tool for conservation, translocation of animals can spread infectious agents to new areas or expose animals to pathogens in their new environment. Mycobacterium avium subsp. paratuberculosis, a bacterium that causes chronic enteritis in ruminants, is among the pathogens of potential concern for wood bison management and conservation. In order to inform translocation decisions, our objectives were to determine the M. avium subsp. paratuberculosis infection status of wood bison herds in Canada and to culture and genetically characterize the infective strain(s). We tested fecal samples from bison (n = 267) in nine herds using direct PCR for three M. avium subsp. paratuberculosis-specific genetic targets with different copy numbers within the M. avium subsp. paratuberculosis genome. Restriction enzyme analysis (REA) and sequencing of IS1311 were performed on seven samples from five different herds. We also evaluated a panel of different culture conditions for their ability to support M. avium subsp. paratuberculosis growth from feces and tissues of direct-PCR-positive animals. Eighty-one fecal samples (30%) tested positive using direct IS900 PCR, with positive samples from all nine herds; of these, 75% and 21% were also positive using ISMAP02 and F57, respectively. None of the culture conditions supported the growth of M. avium subsp. paratuberculosis from PCR-positive samples. IS1311 REA and sequencing indicate that at least two different M. avium subsp. paratuberculosis strain types exist in Canadian wood bison. The presence of different M. avium subsp. paratuberculosis strains among wood bison herds should be considered in the planning of translocations. PMID:23686265

  5. Genome Sequence of Staphylococcus equorum subsp. equorum Mu2, Isolated from a French Smear-Ripened Cheese

    PubMed Central

    Loux, Valentin; Bento, Pascal; Gibrat, Jean-François; Straub, Cécile; Bonnarme, Pascal; Landaud, Sophie; Monnet, Christophe

    2012-01-01

    Staphylococcus equorum subsp. equorum is a member of the coagulase-negative staphylococcus group and is frequently isolated from fermented food products and from food-processing environments. It contributes to the formation of aroma compounds during the ripening of fermented foods, especially cheeses and sausages. Here, we report the draft genome sequence of Staphylococcus equorum subsp. equorum Mu2 to provide insights into its physiology and compare it with other Staphylococcus species. PMID:22933766

  6. Virulence and Immunity Orchestrated by the Global Gene Regulator sigL in Mycobacterium avium subsp. paratuberculosis

    PubMed Central

    Ghosh, Pallab; Steinberg, Howard

    2014-01-01

    Mycobacterium avium subsp. paratuberculosis causes Johne's disease in ruminants, a chronic enteric disease responsible for severe economic losses in the dairy industry. Global gene regulators, including sigma factors are important in regulating mycobacterial virulence. However, the biological significance of such regulators in M. avium subsp. paratuberculosis rremains elusive. To better decipher the role of sigma factors in M. avium subsp. paratuberculosis pathogenesis, we targeted a key sigma factor gene, sigL, activated in mycobacterium-infected macrophages. We interrogated an M. avium subsp. paratuberculosis ?sigL mutant against a selected list of stressors that mimic the host microenvironments. Our data showed that sigL was important in maintaining bacterial survival under such stress conditions. Survival levels further reflected the inability of the ?sigL mutant to persist inside the macrophage microenvironments. Additionally, mouse infection studies suggested a substantial role for sigL in M. avium subsp. paratuberculosis virulence, as indicated by the significant attenuation of the ?sigL-deficient mutant compared to the parental strain. More importantly, when the sigL mutant was tested for its vaccine potential, protective immunity was generated in a vaccine/challenge model of murine paratuberculosis. Overall, our study highlights critical role of sigL in the pathogenesis and immunity of M. avium subsp. paratuberculosis infection, a potential role that could be shared by similar proteins in other intracellular pathogens. PMID:24799632

  7. Local adaptations to frost in marginal and central populations of the dominant forest tree Fagus sylvatica L. as affected by temperature and extreme drought in common garden experiments

    PubMed Central

    Kreyling, Juergen; Buhk, Constanze; Backhaus, Sabrina; Hallinger, Martin; Huber, Gerhard; Huber, Lukas; Jentsch, Anke; Konnert, Monika; Thiel, Daniel; Wilmking, Martin; Beierkuhnlein, Carl

    2014-01-01

    Local adaptations to environmental conditions are of high ecological importance as they determine distribution ranges and likely affect species responses to climate change. Increased environmental stress (warming, extreme drought) due to climate change in combination with decreased genetic mixing due to isolation may lead to stronger local adaptations of geographically marginal than central populations. We experimentally observed local adaptations of three marginal and four central populations of Fagus sylvaticaL., the dominant native forest tree, to frost over winter and in spring (late frost). We determined frost hardiness of buds and roots by the relative electrolyte leakage in two common garden experiments. The experiment at the cold site included a continuous warming treatment; the experiment at the warm site included a preceding summer drought manipulation. In both experiments, we found evidence for local adaptation to frost, with stronger signs of local adaptation in marginal populations. Winter frost killed many of the potted individuals at the cold site, with higher survival in the warming treatment and in those populations originating from colder environments. However, we found no difference in winter frost tolerance of buds among populations, implying that bud survival was not the main cue for mortality. Bud late frost tolerance in April differed between populations at the warm site, mainly because of phenological differences in bud break. Increased spring frost tolerance of plants which had experienced drought stress in the preceding summer could also be explained by shifts in phenology. Stronger local adaptations to climate in geographically marginal than central populations imply the potential for adaptation to climate at range edges. In times of climate change, however, it needs to be tested whether locally adapted populations at range margins can successfully adapt further to changing conditions. PMID:25035801

  8. Ectomycorrhizal Communities on the Roots of Two Beech (Fagus sylvatica) Populations from Contrasting Climates Differ in Nitrogen Acquisition in a Common Environment.

    PubMed

    Leberecht, Martin; Dannenmann, Michael; Gschwendtner, Silvia; Bilela, Silvija; Meier, Rudolf; Simon, Judy; Rennenberg, Heinz; Schloter, Michael; Polle, Andrea

    2015-09-01

    Beech (Fagus sylvatica), a dominant forest species in Central Europe, competes for nitrogen with soil microbes and suffers from N limitation under dry conditions. We hypothesized that ectomycorrhizal communities and the free-living rhizosphere microbes from beech trees from sites with two contrasting climatic conditions exhibit differences in N acquisition that contribute to differences in host N uptake and are related to differences in host belowground carbon allocation. To test these hypotheses, young trees from the natural regeneration of two genetically similar populations, one from dryer conditions (located in an area with a southwest exposure [SW trees]) and the other from a cooler, moist climate (located in an area with a northeast exposure [NE trees]), were transplanted into a homogeneous substrate in the same environment and labeled with (13)CO2 and (15)NH4 (+). Free-living rhizosphere microbes were characterized by marker genes for the N cycle, but no differences between the rhizospheres of SW or NE trees were found. Lower (15)N enrichment was found in the ectomycorrhizal communities of the NE tree communities than the SW tree communities, whereas no significant differences in (15)N enrichment were observed for nonmycorrhizal root tips of SW and NE trees. Neither the ectomycorrhizal communities nor the nonmycorrhizal root tips originating from NE and SW trees showed differences in (13)C signatures. Because the level of (15)N accumulation in fine roots and the amount transferred to leaves were lower in NE trees than SW trees, our data support the suggestion that the ectomycorrhizal community influences N transfer to its host and demonstrate that the fungal community from the dry condition was more efficient in N acquisition when environmental constraints were relieved. These findings highlight the importance of adapted ectomycorrhizal communities for forest nutrition in a changing climate. PMID:26092464

  9. The production, localization and spreading of reactive oxygen species contributes to the low vitality of long-term stored common beech (Fagus sylvatica L.) seeds.

    PubMed

    Ratajczak, Ewelina; Ma?ecka, Arleta; Bagniewska-Zadworna, Agnieszka; Kalemba, Ewa Marzena

    2015-02-01

    The common beech (Fagus sylvatica L.) is propagated by seeds, but the seed set is irregular with five to ten years in between crops. It is therefore necessary to store the seeds. However, beech seeds lose germinability during long-term storage. In this study, beech seeds were stored at -10°C under controlled conditions for 2, 5, 8, 11 and 13 years. Our results show that beech seeds lose germinability during storage in proportion to the duration of storage. The decrease in germinability correlated with increased electrolyte leakage and accumulation of superoxide anion radicals, hydrogen peroxide and hydroxyl radicals. Furthermore, a strong positive correlation was observed among the releases of superoxide anion radicals, hydrogen peroxide and hydroxyl radicals. In situ localization showed that superoxide anion radicals and hydrogen peroxide were first detectable in root cap cells. When the seed storage time was extended, the reactive oxygen species fluorescence expanded to more areas of the radicle, reaching the root apical meristem. A storage time-dependent decrease in catalase activity, observed in both embryonic axes and cotyledons, was also positively correlated with germinability. DNA fragmentation was observed in beech seeds during storage and occurred predominantly in embryonic axes stored for 5 years and more. Altogether, these results suggest that the loss of germinability in beech seeds during long-term storage depends on several factors, including strong of reactive oxygen species accumulation accompanied by reduced catalase activity as well as membrane injury and DNA alternations, which may be aging-related and ROS-derived. We suggest that the accumulating reactive oxygen species that spread to the root apical meristem are key factors that affect seed germinability after long-term storage. PMID:25462977

  10. Within-Population Genetic Structure in Beech (Fagus sylvatica L.) Stands Characterized by Different Disturbance Histories: Does Forest Management Simplify Population Substructure?

    PubMed Central

    Piotti, Andrea; Leonardi, Stefano; Heuertz, Myriam; Buiteveld, Joukje; Geburek, Thomas; Gerber, Sophie; Kramer, Koen; Vettori, Cristina; Vendramin, Giovanni Giuseppe

    2013-01-01

    The fine-scale assessment of both spatially and non-spatially distributed genetic variation is crucial to preserve forest genetic resources through appropriate forest management. Cryptic within-population genetic structure may be more common than previously thought in forest tree populations, which has strong implications for the potential of forests to adapt to environmental change. The present study was aimed at comparing within-population genetic structure in European beech (Fagus sylvatica L.) plots experiencing different disturbance levels. Five plot pairs made up by disturbed and undisturbed plots having the same biogeographic history were sampled throughout Europe. Overall, 1298 individuals were analyzed using four highly polymorphic nuclear microsatellite markers (SSRs). Bayesian clustering within plots identified 3 to 11 genetic clusters (within-plot ?ST ranged from 0.025 to 0.124). The proportion of within-population genetic variation due to genetic substructuring (FCluPlot?=?0.067) was higher than the differentiation among the 10 plots (FPlotTot?=?0.045). Focusing on the comparison between managed and unmanaged plots, disturbance mostly explains differences in the complexity of within-population genetic structure, determining a reduction of the number of genetic clusters present in a standardized area. Our results show that: i) genetic substructuring needs to be investigated when studying the within-population genetic structure in forest tree populations, and ii) indices describing subtle characteristics of the within-population genetic structure are good candidates for providing early signals of the consequences of forest management, and of disturbance events in general. PMID:24039930

  11. Light and competition gradients fail to explain the coexistence of shade-tolerant Fagus sylvatica and shade-intermediate Quercus petraea seedlings

    PubMed Central

    Van Couwenberghe, Rosalinde; Gégout, Jean-Claude; Lacombe, Eric; Collet, Catherine

    2013-01-01

    Background and Aims The coexistence of forest tree species has often been linked to differences among species in terms of their response to light availability during the regeneration stage. From this perspective, species coexistence results from growth–growth or mortality–growth trade-offs along spatial light gradients. Experimental evidence of growth–growth trade-offs in natural conditions is sparse due to various confounding factors that potentially hinder the relationship. This study examined growth hierarchies along light gradients between two tree species with contrasting shade tolerance by controlling potential confounding factors such as seedling size, seedling status, seedling density and species composition. Methods Natural regenerated shade-tolerant Fagus sylvatica and shade-intermediate Quercus petraea seedlings were used, and growth rankings over a 4-year period were compared in 8- to 10-year-old tree seedlings. Key results No rank reversal occurs between the two species along the light gradient, or along the density, mixture or seedling size gradients. The shade-tolerant species was always the more competitive of the two. Pronounced effects of initial size on seedling growth were observed, whereas the effects of light and competition by neighbours were of secondary importance. The paramount effect of size, which results from the asymmetric nature of interseedling competition, gives a strong advantage to tall seedlings over the long term. Conclusions This study extends previous efforts to identify potential drivers of rank reversals in young tree mixtures. It does not support the classical assumption that spatial heterogeneity in canopy opening explains the coexistence of the two species studied. It suggests that spatial variation in local size hierarchies among seedlings that may be caused by seedling emergence time or seedling initial performance is the main driver of the dynamics of these mixed stands. PMID:24036670

  12. Patterns of late spring frost leaf damage and recovery in a European beech (Fagus sylvatica L.) stand in south-eastern Germany based on repeated digital photographs

    PubMed Central

    Menzel, Annette; Helm, Raimund; Zang, Christian

    2015-01-01

    Damage by late spring frost is a risk deciduous trees have to cope with in order to optimize the length of their growing season. The timing of spring phenological development plays a crucial role, not only at the species level, but also at the population and individual level, since fresh new leaves are especially vulnerable. For the pronounced late spring frost in May 2011 in Germany, we studied the individual leaf development of 35 deciduous trees (mainly European beech Fagus sylvatica L.) at a mountainous forest site in the Bayerischer Wald National Park using repeated digital photographs. Analyses of the time series of greenness by a novel Bayesian multiple change point approach mostly revealed five change points which almost perfectly matched the expected break points in leaf development: (i) start of the first greening between day of the year (DOY) 108–119 (mean 113), (ii) end of greening, and (iii) visible frost damage after the frost on the night of May 3rd/4th (DOY 123/124), (iv) re-sprouting 19–38 days after the frost, and (v) full maturity around DOY 178 (166–184) when all beech crowns had fully recovered. Since frost damage was nearly 100%, individual susceptibility did not depend on the timing of first spring leaf unfolding. However, we could identify significant patterns in fitness linked to an earlier start of leaf unfolding. Those individuals that had an earlier start of greening during the first flushing period had a shorter period of recovery and started the second greening earlier. Thus, phenological timing triggered the speed of recovery from such an extreme event. The maximum greenness achieved, however, did not vary with leaf unfolding dates. Two mountain ashes (Sorbus aucuparia L.) were not affected by the low temperatures of -5°C. Time series analysis of webcam pictures can thus improve process-based knowledge and provide valuable insights into the link between phenological variation, late spring frost damage, and recovery within one stand. PMID:25759707

  13. Effects of long-term exposure to ammonium sulfate particles on growth and gas exchange rates of Fagus crenata, Castanopsis sieboldii, Larix kaempferi and Cryptomeria japonica seedlings

    NASA Astrophysics Data System (ADS)

    Yamaguchi, Masahiro; Otani, Yoko; Li, Peiran; Nagao, Hiroshi; Lenggoro, I. Wuled; Ishida, Atsushi; Yazaki, Kenichi; Noguchi, Kyotaro; Nakaba, Satoshi; Yamane, Kenichi; Kuroda, Katsushi; Sano, Yuzou; Funada, Ryo; Izuta, Takeshi

    2014-11-01

    To clarify the effects of long-term exposure to ammonium sulfate (AS) particles on growth and physiological functions of forest tree species, seedlings of Fagus crenata, Castanopsis sieboldii, Larix kaempferi and Cryptomeria japonica were exposed to submicron-size AS particles during two growing seasons from 3 June 2011 to 8 October 2012. The mean sulfate concentration in PM2.5 increased during the exposure inside the chamber in 2011 and 2012 by 2.73 and 4.32 ?g SO42- m-3, respectively. No significant effects of exposure to AS particles were detected on the whole-plant dry mass of the seedlings. These results indicate that the exposure to submicrometer AS particles at the ambient level for two growing seasons did not significantly affect the growth of the seedlings. No significant effects of exposure to AS particles were found on the net photosynthetic rate in the leaves or needles of F. crenata, C. sieboldii and L. kaempferi seedlings. Also, in the previous-year needles of C. japonica seedlings, exposure to AS particles significantly reduced the net photosynthetic rate, which may be caused by the reduction in the concentration of ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco). On the contrary, in current-year needles of C. japonica seedlings, net photosynthetic rate significantly increased with exposure to AS particles, which may be the result of increases in stomatal conductance and concentrations of Rubisco and chlorophyll. Furthermore, exposure to AS particles correlated with an increase in concentrations of NH4+, free amino acid and total soluble protein, suggesting that AS particles may be deliquesced, absorbed into the leaves and metabolized into amino acid and protein. These results suggest that net photosynthesis in the needles of C. japonica is relatively sensitive to submicron-size AS particles as compared with the other three tree species.

  14. The importance of atmospheric deposition, charge and atomic mass to the dynamics of minor and rare elements in developing, ageing, and wilted leaves of beech (Fagus sylvatica L.).

    PubMed

    Tyler, Germund; Olsson, Tommy

    2006-10-01

    The amounts of sixty elements in developing, maturing, senescent and wilting leaves, and in the wintering dead leaves attached to the branches, are reported for a beech (Fagus sylvatica) forest on mor Podzol in south Sweden, a site with no local sources of pollution or geological anomalies. The amounts (contents per leaf) of K (potassium), Rb (rubidium), Cs (caesium), Cu (copper) and P (phosphorus) were highest in young leaves, decreasing throughout the growing season and usually in the subsequent winter. The entirely opposite pattern with a continuous, mostly even increase of the amounts was measured with Be (beryllium), Ba (barium), Hg (mercury), Al (aluminium), Tl (thallium), Pb (lead), Bi (bismuth), V (vanadium), W (tungsten), As (arsenic), Sb (antimony), and Se (selenium). Amounts of rare-earth elements and some transition metals, such as Co (cobalt), Ti (titanium), and the actinides Th (thorium) and U (uranium) were more stable during the growing season, after an initial increase in early summer, but increased greatly in the winter. This winter increase in dead attached leaves has to be accounted for by uptake from long-distance transported constituents in dry and wet deposition. It was similar to deposition rate estimates using moss carpets from the same locality. A passive uptake was positively related to ionic charge and atomic mass. However, the amounts of several, mainly non-essential elements, such as Ni (nickel), Sc (scandium), Zr (zirconium), Cr (chromium), Ag (silver), and Cd (cadmium) were not much lower in the young or maturing leaves than in the wintered dead leaves of this deciduous (hardwood) forest and a proportion apparently originated from internal translocation in the trees. Seasonal fluxes or cycling of many of the scarce or rare elements reported here have never been studied before in forest ecosystems. PMID:16603223

  15. Biotic and Abiotic Factors Controlling Respiration Rates of Above- and Belowground Woody Debris of Fagus crenata and Quercus crispula in Japan

    PubMed Central

    Jomura, Mayuko; Akashi, Yuhei; Itoh, Hiromu; Yuki, Risa; Sakai, Yoshimi; Maruyama, Yutaka

    2015-01-01

    As a large, long-term pool and source of carbon and nutrients, woody litter is an important component of forest ecosystems. The objective of this study was to estimate the effect of the factors that regulate the rate of decomposition of coarse and fine woody debris (CFWD) of dominant tree species in a cool-temperate forest in Japan. Respiration rates of dead stems, branches, and coarse and fine roots of Fagus crenata and Quercus crispula felled 4 years prior obtained in situ ranged from 20.9 to 500.1 mg CO2 [kg dry wood]–1 h–1 in a one-time measurement in summer. Respiration rate had a significant negative relationship with diameter; in particular, that of a sample of Q. crispula with a diameter of >15 cm and substantial heartwood was low. It also had a significant positive relationship with moisture content. The explanatory variables diameter, [N], wood density, and moisture content were interrelated. The most parsimonious path model showed 14 significant correlations among 8 factors and respiration. Diameter and [C] had large negative direct effects on CFWD respiration rate, and moisture content and species had medium positive direct effects. [N] and temperature did not have direct or indirect effects, and position and wood density had indirect effects. The model revealed some interrelationships between controlling factors. We discussed the influence of the direct effects of explanatory variables and the influence especially of species and position. We speculate that the small R2 value of the most parsimonious model was probably due to the omission of microbial biomass and activity. These direct and indirect effects and interrelationships between explanatory variables could be used to develop a process-based CFWD decomposition model. PMID:26658727

  16. Draft Genome Sequence of Lactobacillus delbrueckii subsp. bulgaricus CRL871, a Folate-Producing Strain Isolated from a Northwestern Argentinian Yogurt.

    PubMed

    Laiño, Jonathan Emiliano; Hebert, Elvira María; Savoy de Giori, Graciela; LeBlanc, Jean Guy

    2015-01-01

    Lactobacillus delbrueckii subsp. bulgaricus CRL871 is the first strain of L. delbrueckii subsp. bulgaricus reported as a folate-producing strain. We report the draft genome sequence of L. delbrueckii subsp. bulgaricus CRL871 (2,063,981 bp, G+C content of 49.1%). This strain is of great biotechnological importance to the dairy industry because it constitutes an alternative to folic acid fortification. PMID:26112792

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

    PubMed Central

    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

  18. Draft Genome Sequence of Xylella fastidiosa subsp. multiplex Strain Griffin-1 from Quercus rubra in Georgia.

    PubMed

    Chen, Jianchi; Huang, Hong; Chang, Chung-Jan; Stenger, Drake C

    2013-01-01

    The draft genome sequence of Xylella fastidiosa subsp. multiplex strain Griffin-1, isolated from a red oak tree (Quercus rubra) in Georgia, is reported here. The bacterium has a genome size of 2,387,314 bp, with a G+C content of 51.7%. The Griffin-1 strain genome contains 2,903 predicted open reading frames and 50 RNA genes. PMID:24115539

  19. Cytotoxic and antibacterial labdane-type diterpenes from the aerial parts of Cistus incanus subsp. creticus.

    PubMed

    Chinou, I; Demetzos, C; Harvala, C; Roussakis, C; Verbist, J F

    1994-02-01

    Seven labdane-type diterpenoids were isolated from the leaves of Cistus incanus subsp. creticus; their structures were established by spectroscopic means. All compounds were tested in vitro for their cytotoxicity against three cell line systems: KB, P-388; and NSCLC-N6. Their antibacterial and antifungal activities were tested against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosae, Enterobacter cloacae, Escherichia coli, Klebsiella pneumonia, Torulopsis glabrata, Saccharomyces cerevisiae, and Candida albicans as well. PMID:8134413

  20. Novel Phytases from Bifidobacterium pseudocatenulatum ATCC 27919 and Bifidobacterium longum subsp. infantis ATCC 15697

    PubMed Central

    Tamayo-Ramos, Juan Antonio; Sanz-Penella, Juan Mario; Yebra, María J.

    2012-01-01

    Two novel phytases have been characterized from Bifidobacterium pseudocatenulatum and Bifidobacterium longum subsp. infantis. The enzymes belong to a new subclass within the histidine acid phytases, are highly specific for the hydrolysis of phytate, and render myo-inositol triphosphate as the final hydrolysis product. They represent the first phytases characterized from this group of probiotic microorganisms, opening the possibilities for their use in the processing of high-phytate-content foods. PMID:22582052

  1. Novel phytases from Bifidobacterium pseudocatenulatum ATCC 27919 and Bifidobacterium longum subsp. infantis ATCC 15697.

    PubMed

    Tamayo-Ramos, Juan Antonio; Sanz-Penella, Juan Mario; Yebra, María J; Monedero, Vicente; Haros, Monika

    2012-07-01

    Two novel phytases have been characterized from Bifidobacterium pseudocatenulatum and Bifidobacterium longum subsp. infantis. The enzymes belong to a new subclass within the histidine acid phytases, are highly specific for the hydrolysis of phytate, and render myo-inositol triphosphate as the final hydrolysis product. They represent the first phytases characterized from this group of probiotic microorganisms, opening the possibilities for their use in the processing of high-phytate-content foods. PMID:22582052

  2. Optimization of Hexadecylpyridinium Chloride Decontamination for Culture of Mycobacterium avium subsp. paratuberculosis from Milk

    PubMed Central

    Bradner, L.; Robbe-Austerman, S.; Beitz, D. C.

    2013-01-01

    A protocol was optimized for the isolation of Mycobacterium avium subsp. paratuberculosis (MAP) from milk and colostrum, with parameters including chemical decontamination, antibiotics, and different culture media. This study demonstrates that the efficiency of MAP recovery from milk is highly dependent upon the culturing protocol, and such protocols should be optimized to ensure that low concentrations of MAP in milk can be detected. PMID:23426920

  3. Hare-to-Human Transmission of Francisella tularensis subsp. holarctica, Germany

    PubMed Central

    Otto, Peter; Kohlmann, Rebekka; Müller, Wolfgang; Julich, Sandra; Geis, Gabriele; Gatermann, Sören G.; Peters, Martin; Wolf, Peter Johannes; Karlsson, Edvin; Forsman, Mats; Myrtennäs, Kerstin

    2015-01-01

    In November 2012, a group of 7 persons who participated in a hare hunt in North Rhine-Westphalia, Germany, acquired tularemia. Two F. tularensis subsp. holarctica isolates were cultivated from human and hare biopsy material. Both isolates belonged to the FTN002–00 genetic subclade (derived for single nucleotide polymorphisms B.10 and B.18), thus indicating likely hare-to-human transmission. PMID:25531286

  4. Avian wildlife reservoir of Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. in Norway.

    PubMed Central

    Kapperud, G; Rosef, O

    1983-01-01

    Cloacal swabs from 540 wild-living birds were cultured for Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. The carrier rates detected were as follows: C. fetus subsp. jejuni, 28.4%; Yersinia spp., 1.2%; and Salmonella spp., 0.8%. All birds were apparently healthy when captured. C. fetus subsp. jejuni was isolated from 11 of the 40 bird species examined. Among birds inhabiting the city of Oslo, the highest isolation rate was found in crows (Corvus corone cornix) (89.8%), followed by gulls (Larus spp.) (50.0%) and domestic pigeons (Columba livia domesticus) (4.2%). The gulls and crows scavenge on refuse dumps. High carrier rates were also detected among the following birds from nonurban, coastal areas: puffin (Fratercula arctica) (51.3%), common tern (Sterna hirundo) (5.6%), common gull (Larus canus) (18.9%), black-headed gull (Larus ridibundus) (13.2%), and herring gull (Larus argentatus) (4.2%). The list of species harboring C. fetus subsp. jejuni also includes the Ural owl (Strix uralensis), goldeneye (Bucephala clangula), and reed bunting (Emberiza schoeniclus). The following five Yersinia strains were isolated: Y. kristensenii (two strains), Y. intermedia (two strains), and "Yersinia X2" (one strain). Four strains belonging to the genus Salmonella were isolated from three different species of gulls. These isolates were identified as S. typhimurium, S. indiana, and S. djugu. The results indicate that campylobacters are a normal component of the intestinal flora in several bird species, whereas Salmonella and Yersinia carriers are more sporadic. PMID:6338824

  5. Avian wildlife reservoir of Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. in Norway.

    PubMed

    Kapperud, G; Rosef, O

    1983-02-01

    Cloacal swabs from 540 wild-living birds were cultured for Campylobacter fetus subsp. jejuni, Yersinia spp., and Salmonella spp. The carrier rates detected were as follows: C. fetus subsp. jejuni, 28.4%; Yersinia spp., 1.2%; and Salmonella spp., 0.8%. All birds were apparently healthy when captured. C. fetus subsp. jejuni was isolated from 11 of the 40 bird species examined. Among birds inhabiting the city of Oslo, the highest isolation rate was found in crows (Corvus corone cornix) (89.8%), followed by gulls (Larus spp.) (50.0%) and domestic pigeons (Columba livia domesticus) (4.2%). The gulls and crows scavenge on refuse dumps. High carrier rates were also detected among the following birds from nonurban, coastal areas: puffin (Fratercula arctica) (51.3%), common tern (Sterna hirundo) (5.6%), common gull (Larus canus) (18.9%), black-headed gull (Larus ridibundus) (13.2%), and herring gull (Larus argentatus) (4.2%). The list of species harboring C. fetus subsp. jejuni also includes the Ural owl (Strix uralensis), goldeneye (Bucephala clangula), and reed bunting (Emberiza schoeniclus). The following five Yersinia strains were isolated: Y. kristensenii (two strains), Y. intermedia (two strains), and "Yersinia X2" (one strain). Four strains belonging to the genus Salmonella were isolated from three different species of gulls. These isolates were identified as S. typhimurium, S. indiana, and S. djugu. The results indicate that campylobacters are a normal component of the intestinal flora in several bird species, whereas Salmonella and Yersinia carriers are more sporadic. PMID:6338824

  6. Growth and Energy Generation by Lactococcus lactis subsp. lactis biovar diacetylactis during Citrate Metabolism.

    PubMed

    Hugenholtz, J; Perdon, L; Abee, T

    1993-12-01

    Growth of Lactococcus lactis subsp. lactis biovar diacetylactis was observed on media with citrate as the only energy source. At pH 5.6, steady state was achieved in a chemostat on a citrate-containing medium in the absence of a carbohydrate. Under these conditions, pyruvate, acetate, and some acetoin and butanediol were the main fermentation products. This indicated that energy was conserved in L. lactis subsp. lactis biovar diacetylactis during citrate metabolism and presumably during the conversion of citrate into pyruvate. The presumed energy-conserving step, decarboxylation of oxaloacetate, was studied in detail. Oxaloacetate decarboxylase was purified to homogeneity and characterized. The enzyme has a native molecular mass of approximately 300 kDa and consists of three subunits of 52, 34, and 12 kDa. The enzyme is apparently not sodium dependent and does not contain a biotin moiety, and it seems to be different from the energy-generating oxaloacetate decarboxylase from Klebsiella pneumoniae. Energy-depleted L. lactis subsp. lactis biovar diacetylactis cells generated a membrane potential and a pH gradient immediately upon addition of citrate, whereas ATP formation was slow and limited. In contrast, lactose energization resulted in rapid ATP formation and gradual generation of a proton motive force. These data were confirmed during studies on amino acid uptake. alpha-Aminoisobutyrate uptake was rapid but glutamate uptake was slow in citrate-energized cells, whereas lactose-energized cells showed the reverse tendency. These data suggest that, in L. lactis subsp. lactis bv. diacetylactis, a proton motive force could be generated during citrate metabolism as a result of electrogenic citrate uptake or citrate/product exchange together with proton consumption by the intracellular oxaloacetate decarboxylase. PMID:16349120

  7. Growth and Energy Generation by Lactococcus lactis subsp. lactis biovar diacetylactis during Citrate Metabolism

    PubMed Central

    Hugenholtz, Jeroen; Perdon, Leo; Abee, Tjakko

    1993-01-01

    Growth of Lactococcus lactis subsp. lactis biovar diacetylactis was observed on media with citrate as the only energy source. At pH 5.6, steady state was achieved in a chemostat on a citrate-containing medium in the absence of a carbohydrate. Under these conditions, pyruvate, acetate, and some acetoin and butanediol were the main fermentation products. This indicated that energy was conserved in L. lactis subsp. lactis biovar diacetylactis during citrate metabolism and presumably during the conversion of citrate into pyruvate. The presumed energy-conserving step, decarboxylation of oxaloacetate, was studied in detail. Oxaloacetate decarboxylase was purified to homogeneity and characterized. The enzyme has a native molecular mass of approximately 300 kDa and consists of three subunits of 52, 34, and 12 kDa. The enzyme is apparently not sodium dependent and does not contain a biotin moiety, and it seems to be different from the energy-generating oxaloacetate decarboxylase from Klebsiella pneumoniae. Energy-depleted L. lactis subsp. lactis biovar diacetylactis cells generated a membrane potential and a pH gradient immediately upon addition of citrate, whereas ATP formation was slow and limited. In contrast, lactose energization resulted in rapid ATP formation and gradual generation of a proton motive force. These data were confirmed during studies on amino acid uptake. ?-Aminoisobutyrate uptake was rapid but glutamate uptake was slow in citrate-energized cells, whereas lactose-energized cells showed the reverse tendency. These data suggest that, in L. lactis subsp. lactis bv. diacetylactis, a proton motive force could be generated during citrate metabolism as a result of electrogenic citrate uptake or citrate/product exchange together with proton consumption by the intracellular oxaloacetate decarboxylase. Images PMID:16349120

  8. Hare-to-human transmission of Francisella tularensis subsp. holarctica, Germany.

    PubMed

    Otto, Peter; Kohlmann, Rebekka; Müller, Wolfgang; Julich, Sandra; Geis, Gabriele; Gatermann, Sören G; Peters, Martin; Wolf, Peter Johannes; Karlsson, Edvin; Forsman, Mats; Myrtennäs, Kerstin; Tomaso, Herbert

    2015-01-01

    In November 2012, a group of 7 persons who participated in a hare hunt in North Rhine-Westphalia, Germany, acquired tularemia. Two F. tularensis subsp. holarctica isolates were cultivated from human and hare biopsy material. Both isolates belonged to the FTN002-00 genetic subclade (derived for single nucleotide polymorphisms B.10 and B.18), thus indicating likely hare-to-human transmission. PMID:25531286

  9. Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae

    PubMed Central

    Yu, Mingyan; O'Brien, Elizabeth; Heller, Martin; Nepper, Julia F.; Weibel, Douglas B.; Gluecks, Ilona; Younan, Mario; Frey, Joachim; Falquet, Laurent; Jores, Joerg

    2015-01-01

    Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of Mycoplasma infection, including cultivation, serological assays, and PCR, are time-consuming and require fully equipped stationary laboratories, which make them incompatible with testing in the resource-poor settings that are most relevant to this disease. We report a rapid, specific, and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for the detection of M. capricolum subsp. capripneumoniae. We developed the assay using a specific target sequence in M. capricolum subsp. capripneumoniae, as found in the genome sequence of the field strain ILRI181 and the type strain F38 and that was further evidenced in 10 field strains from different geographical regions. Detection limits corresponding to 5 × 103 and 5 × 104 cells/ml were obtained using genomic DNA and bacterial culture from M. capricolum subsp. capripneumoniae strain ILRI181, while no amplification was obtained from 71 related Mycoplasma isolates or from the Acholeplasma or the Pasteurella isolates, demonstrating a high degree of specificity. The assay produces a fluorescent signal within 15 to 20 min and worked well using pleural fluid obtained directly from CCPP-positive animals without prior DNA extraction. We demonstrate that the diagnosis of CCPP can be achieved, with a short sample preparation time and a simple read-out device that can be powered by a car battery, in <45 min in a simulated field setting. PMID:26085615

  10. The importance of arbuscular mycorrhiza for Cyclamen purpurascens subsp. immaculatum endemic in Slovakia.

    PubMed

    Rydlová, Jana; Sýkorová, Zuzana; Slavíková, Renata; Turis, Peter

    2015-11-01

    At present, there is no relevant information on arbuscular mycorrhiza and the effect of the symbiosis on the growth of wild populations of cyclamens. To fill this gap, two populations of Cyclamen purpurascens subsp. immaculatum, endemic in Nízke Tatry (NT) mountains and Ve?ká Fatra (VF) mountains, Slovakia, were studied in situ as well as in a greenhouse pot experiment. For both populations, mycorrhizal root colonization of native plants was assessed, and mycorrhizal inoculation potential (MIP) of the soils at the two sites was determined in 3 consecutive years. In the greenhouse experiment, the growth response of cyclamens to cross-inoculation with arbuscular mycorrhizal fungi (AMF) was tested: plants from both sites were grown in their native soils and inoculated with a Septoglomus constrictum isolate originating either from the same or from the other plant locality. Although the MIP of soil at the NT site was significantly higher than at the VF site, the level of AMF root colonization of C. purpurascens subsp. immaculatum plants in the field did not significantly differ between the two localities. In the greenhouse experiment, inoculation with AMF generally accelerated cyclamen growth and significantly increased all growth parameters (shoot dry weight, leaf number and area, number of flowers, tuber, and root dry weight) and P uptake. The two populations of C. purpurascens subsp. immaculatum grown in their native soils, however, differed in their response to inoculation. The mycorrhizal growth response of NT plants was one-order higher compared to VF plants, and all their measured growth parameters were stimulated regardless of the fungal isolates' origin. In the VF plants, only the non-native (NT originating) isolate showed a significant positive effect on several growth traits. It can be concluded that mycorrhiza significantly increased fitness of C. purpurascens subsp. immaculatum, despite the differences between plant populations, implying that AMF symbionts should be taken into account in conservation programs of this endemic plant. PMID:25720737

  11. Immunoreactivity of the Mycobacterium avium subsp. paratuberculosis 19-kDa lipoprotein

    PubMed Central

    Huntley, Jason FJ; Stabel, Judith R; Bannantine, John P

    2005-01-01

    Background The Mycobacterium tuberculosis 19-kDa lipoprotein has been reported to stimulate both T and B cell responses as well as induce a number of Th1 cytokines. In order to evaluate the Mycobacterium avium subsp. paratuberculosis (M. avium subsp. paratuberculosis) 19-kDa lipoprotein as an immunomodulator in cattle with Johne's disease, the gene encoding the 19-kDa protein (MAP0261c) was analyzed. Results MAP0261c is conserved in mycobacteria, showing a 95% amino acid identity in M. avium subspecies avium, 84% in M. intracellulare and 76% in M. bovis and M. tuberculosis. MAP0261c was cloned, expressed, and purified as a fusion protein with the maltose-binding protein (MBP-19 kDa) in Escherichia coli. IFN-? production was measured from 21 naturally infected and 9 control cattle after peripheral blood mononuclear cells (PBMCs) were stimulated with a whole cell lysate (WCL) of M. avium subsp. paratuberculosis or the recombinant MBP-19 kDa. Overall, the mean response to MBP-19 kDa was not as strong as the mean response to the WCL. By comparison, cells from control, non-infected cattle did not produce IFN-? after stimulation with either WCL or MBP-19 kDa. To assess the humoral immune response to the 19-kDa protein, sera from cattle with clinical Johne's disease were used in immunoblot analysis. Reactivity to MBP-19 kDa protein, but not MBP alone, was observed in 9 of 14 infected cattle. Antibodies to the 19-kDa protein were not observed in 8 of 9 control cows. Conclusions Collectively, these results demonstrate that while the 19-kDa protein from M. avium subsp. paratuberculosis stimulates a humoral immune response and weak IFN-? production in infected cattle, the elicited responses are not strong enough to be used in a sensitive diagnostic assay. PMID:15663791

  12. Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae.

    PubMed

    Liljander, Anne; Yu, Mingyan; O'Brien, Elizabeth; Heller, Martin; Nepper, Julia F; Weibel, Douglas B; Gluecks, Ilona; Younan, Mario; Frey, Joachim; Falquet, Laurent; Jores, Joerg

    2015-09-01

    Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of Mycoplasma infection, including cultivation, serological assays, and PCR, are time-consuming and require fully equipped stationary laboratories, which make them incompatible with testing in the resource-poor settings that are most relevant to this disease. We report a rapid, specific, and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for the detection of M. capricolum subsp. capripneumoniae. We developed the assay using a specific target sequence in M. capricolum subsp. capripneumoniae, as found in the genome sequence of the field strain ILRI181 and the type strain F38 and that was further evidenced in 10 field strains from different geographical regions. Detection limits corresponding to 5 × 10(3) and 5 × 10(4) cells/ml were obtained using genomic DNA and bacterial culture from M. capricolum subsp. capripneumoniae strain ILRI181, while no amplification was obtained from 71 related Mycoplasma isolates or from the Acholeplasma or the Pasteurella isolates, demonstrating a high degree of specificity. The assay produces a fluorescent signal within 15 to 20 min and worked well using pleural fluid obtained directly from CCPP-positive animals without prior DNA extraction. We demonstrate that the diagnosis of CCPP can be achieved, with a short sample preparation time and a simple read-out device that can be powered by a car battery, in <45 min in a simulated field setting. PMID:26085615

  13. Draft Genome Sequence of Staphylococcus sciuri subsp. sciuri Strain Z8, Isolated from Human Skin

    PubMed Central

    Hu, Xinjun; Zheng, Beiwen; Jiang, Haiyin; Kang, Yi; Cao, Qing; Ning, Huibin

    2015-01-01

    Staphylococcus sciuri subsp. sciuri strain Z8 was isolated from a skin wound infection of a patient with infective endocarditis. To the best of our knowledge, the genome sequence of the species S. sciuri has not been previously studied. The complete genome sequence of strain Z8 includes a genome of 2,620,868 bp (32.43% GC content) without any plasmids. PMID:26205872

  14. Genome differences between Treponema pallidum subsp. pallidum strain Nichols and T. paraluiscuniculi strain Cuniculi A.

    PubMed

    Strouhal, Michal; Smajs, David; Matejková, Petra; Sodergren, Erica; Amin, Anita G; Howell, Jerrilyn K; Norris, Steven J; Weinstock, George M

    2007-12-01

    The genome of Treponema paraluiscuniculi strain Cuniculi A was compared to the genome of the syphilis spirochete Treponema pallidum subsp. pallidum strain Nichols using DNA microarray hybridization, whole-genome fingerprinting, and DNA sequencing. A DNA microarray of T. pallidum subsp. pallidum Nichols containing all 1,039 predicted open reading frame PCR products was used to identify deletions and major sequence changes in the Cuniculi A genome. Using these approaches, deletions, insertions, and prominent sequence changes were found in 38 gene homologs and six intergenic regions of the Cuniculi A genome when it was compared to the genome of T. pallidum subsp. pallidum Nichols. Most of the observed differences were localized in tpr loci and the vicinity of these loci. In addition, 14 other genes were found to contain frameshift mutations resulting in major changes in protein sequences. Analysis of restriction target sites representing 0.34% of the total genome length and DNA sequencing of three PCR products (0.46% of the total genome length) amplified from Cuniculi A chromosomal regions and comparison to the Nichols genome revealed a sequence similarity of 98.6 to 99.3%. These results are consistent with a close genetic relationship among the T. pallidum strains and subspecies and a strong, but relatively divergent connection between the human and rabbit pathogens. PMID:17893135

  15. Molecular cloning and functional analysis of Photobacterium damselae subsp. piscicida haem receptor gene.

    PubMed

    Naka, H; Hirono, I; Aoki, T

    2005-02-01

    A haem receptor gene from Photobacterium damselae subsp. piscicida (formerly known as Pasteurella piscicida) has been cloned, sequenced and analysed for its function. The gene, designated as pph, has an open reading frame consisting of 2154 bp, a predicted 718 amino acid residues and exists as a single copy. It is homologous with the haem receptors of Vibrio anguillarum hupA, V. cholerae hutA, V. mimicus mhuA and V. vulnificus hupA at 32.7, 32.7, 45.6 and 30.9%, respectively, and is highly conserved, consisting of a Phe-Arg-Ala-Pro sequence (FRAP), an iron transport related molecule (TonB) and a Asn-Pron-Asn-Leu sequence (NPNL), binding motifs associated with haem receptors. As a single gene knockout mutant P. damselae subsp. piscicida was able to bind haem in the absence of pph, suggesting that other receptors may be involved in its iron transport system. This study shows that the P. damselae subsp. piscicida pph belongs to the haem receptor family, is conserved and that its iron-binding system may involve more than one receptor. PMID:15705153

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

    PubMed Central

    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

  17. Lymphoproliferative and Gamma Interferon Responses to Stress-Regulated Mycobacterium avium subsp. paratuberculosis Recombinant Proteins

    PubMed Central

    Gurung, Ratna B.; Begg, Douglas J.; Purdie, Auriol C.; de Silva, Kumudika; Bannantine, John P.

    2014-01-01

    Johne's disease in ruminants is a chronic infection of the intestines caused by Mycobacterium avium subsp. paratuberculosis. An important strategy to control disease is early detection, and a potentially efficient method for early detection is measurement of cell-mediated immune responses developed by the host in response to exposure or infection. One method is to measure lymphoproliferation and cytokine release from the host cells when exposed to the organism or parts of the organism. In this study, 10 recombinant M. avium subsp. paratuberculosis proteins known to be upregulated under in vitro stress conditions were evaluated by examining their ability to evoke memory as a result of exposure by vaccination or oral challenge with live Mycobacterium avium subsp. paratuberculosis. Out of 10 proteins, MAP2698c was found to induce higher cell-mediated immune responses in vaccinated and challenged sheep in comparison to healthy controls. The findings suggest that not all stress-regulated proteins have the diagnostic potential to detect cell-mediated immune responses in ovine paratuberculosis. PMID:24695774

  18. Phobalysin, a Small ?-Pore-Forming Toxin of Photobacterium damselae subsp. damselae.

    PubMed

    Rivas, Amable J; von Hoven, Gisela; Neukirch, Claudia; Meyenburg, Martina; Qin, Qianqian; Füser, Sabine; Boller, Klaus; Lemos, Manuel L; Osorio, Carlos R; Husmann, Matthias

    2015-11-01

    Photobacterium damselae subsp. damselae, an important pathogen of marine animals, may also cause septicemia or hyperaggressive necrotizing fasciitis in humans. We previously showed that hemolysin genes are critical for virulence of this organism in mice and fish. In the present study, we characterized the hlyA gene product, a putative small ?-pore-forming toxin, and termed it phobalysin P (PhlyP), for "photobacterial lysin encoded on a plasmid." PhlyP formed stable oligomers and small membrane pores, causing efflux of K(+), with no significant leakage of lactate dehydrogenase but entry of vital dyes. The latter feature distinguished PhlyP from the related Vibrio cholerae cytolysin. Attack by PhlyP provoked a loss of cellular ATP, attenuated translation, and caused profound morphological changes in epithelial cells. In coculture experiments with epithelial cells, Photobacterium damselae subsp. damselae led to rapid hemolysin-dependent membrane permeabilization. Unexpectedly, hemolysins also promoted the association of P. damselae subsp. damselae with epithelial cells. The collective observations of this study suggest that membrane-damaging toxins commonly enhance bacterial adherence. PMID:26303391

  19. Phospholipase A2 Inhibitors Synthesized by Two Entomopathogenic Bacteria, Xenorhabdus nematophila and Photorhabdus temperata subsp. temperata

    PubMed Central

    Seo, Samyeol; Lee, Sunghong; Hong, Yongpyo

    2012-01-01

    The entomopathogenic bacteria Xenorhabdus nematophila and Photorhabdus temperata subsp. temperata suppress insect immune responses by inhibiting the catalytic activity of phospholipase A2 (PLA2), which results in preventing biosynthesis of immune-mediating eicosanoids. This study identified PLA2 inhibitors derived from culture broths of these two bacteria. Both X. nematophila and P. temperata subsp. temperata culture broths possessed significant PLA2-inhibitory activities. Fractionation of these bacterial metabolites in the culture broths using organic solvent and subsequent chromatography purified seven potent PLA2 inhibitors, three of which (benzylideneacetone [BZA], proline-tyrosine [PY], and acetylated phenylalanine-glycine-valine [FGV]) were reported in a previous study. Four other compounds (indole, oxindole, cis-cyclo-PY, and p-hydroxyphenyl propionic acid) were identified and shown to significantly inhibit PLA2. X. nematophila culture broth contained these seven compounds, while P. temperata subsp. temperata culture broth contained three compounds (BZA, acetylated FGV, and cis-cyclo-PY). BZA was detected in the largest amount among these PLA2 compounds in both bacterial culture broths. All seven bacterial metabolites also showed significant inhibitory activities against immune responses, such as phenoloxidase activity and hemocytic nodulation; BZA was the most potent. Finally, this study characterized these seven compounds for their insecticidal activities against the diamondback moth, Plutella xylostella. Even though these compounds showed relatively low toxicities to larvae, they significantly enhanced the pathogenicity of Bacillus thuringiensis. This study reports bacterial-origin PLA2 inhibitors, which would be applicable for developing novel insecticides. PMID:22447611

  20. Domain analysis of lipoprotein LppQ in Mycoplasma mycoides subsp. mycoides SC

    PubMed Central

    Bonvin-Klotz, Laetitia; Kühni-Boghenbor, Kathrin; Kapp, Nadine; Frey, Joachim; Stoffel, Michael H.

    2007-01-01

    The lipoprotein LppQ is the most prominent antigen of Mycoplasma mycoides subsp. mycoides small colony type (SC) during infection of cattle. This pathogen causes contagious bovine pleuropneumonia (CBPP), a devastating disease of considerable socio-economic importance in many countries worldwide. The dominant antigenicity and high specificity for M. mycoides subsp. mycoides SC of lipoprotein LppQ have been exploited for serological diagnosis and for epidemiological investigations of CBPP. Scanning electron microscopy and immunogold labelling were used to provide ultrastructural evidence that LppQ is located to the cell membrane at the outer surface of M. mycoides subsp. mycoides SC. The selectivity and specificity of this method were demonstrated through discriminating localization of extracellular (i.e., in the zone of contact with host cells) vs. integral membrane domains of LppQ. Thus, our findings support the suggestion that the accessible N-terminal domain of LppQ is surface exposed and such surface localization may be implicated in the pathogenesis of CBPP. PMID:17674137

  1. Antigenic and Genetic Characterization of Lipoprotein LppQ from Mycoplasma mycoides subsp. mycoides SC

    PubMed Central

    Abdo, El-Mostafa; Nicolet, Jacques; Frey, Joachim

    2000-01-01

    Lipoprotein LppQ, a predominant 48-kDa antigen, and its corresponding gene, lppQ, were characterized in Mycoplasma mycoides subsp. mycoides SC, the etiological agent of contagious bovine pleuropneumonia. The lppQ gene is specific to M. mycoides subsp. mycoides SC and was found in the type strain and in field strains isolated in Europe, Africa, and Australia, as well as in vaccinal strains. LppQ is encoded as a precursor with a consensus sequence for prokaryotic signal peptidase II and a lipid attachment site. The leader sequence shows significant prominent transmembrane helix structure with a predicted outside-to-inside helix formation capacity. The N-terminal domain of the mature LppQ was shown to be surface exposed. It induced a strong, specific, early, and persistent immune response in naturally and experimentally infected animals. The C-terminal domain of LppQ possesses an integral membrane structure built up of repeated units, rich in hydrophobic and aromatic amino acids, which have a pore formation potential. A recombinant peptide representing the N-terminal domain of LppQ was obtained by site-directed mutagenesis of nine Mycoplasma-specific TGA (Trp) codons into universal TGG (Trp) codons and expression in Escherichia coli hosts. It was used for serodetection of cattle infected with M. mycoides subsp. mycoides SC, in which it was detected postinfection for significantly longer than conventional serological test reactions. PMID:10882657

  2. Characterization of pathogenic vibrios isolated from bivalve hatcheries in Galicia, NW Atlantic coast of Spain. Description of Vibrio tubiashii subsp. europaensis subsp. nov.

    PubMed

    Prado, Susana; Dubert, Javier; Barja, Juan L

    2015-02-01

    The taxonomic position of the bivalve pathogen PP-638 was studied together with five similar isolates. The strains were isolated from flat oyster (Ostrea edulis) and Manila clam (Venerupis philippinarum) cultures during outbreaks of disease in two shellfish hatcheries (Galicia, NW Spain). The pathogenicity, previously established for PP-638, was demonstrated with all isolates and for several bivalve species, including the original hosts. On the basis of phenotypic characterization and 16S rRNA gene sequences, a tight group was defined within the genus Vibrio. Multilocus sequence analysis (MLSA) based on concatenated sequences of the 16S rRNA gene and the five housekeeping genes recA, rpoA, pyrH, gyrB and ftsZ revealed that these strains form a cluster within the Orientalis clade, close to the species Vibrio tubiashii. The results of MLSA, the DDH rate and the phenotypic differences with the type strain of V. tubiashii supported the differentiation of the Galician isolates as a new subspecies within V. tubiashii, for which the name V. tubiashii subsp. europaensis subsp. nov. is proposed (type strain PP-638(T)=CECT 8136(T)=DSM 7349(T)) The emended description of V. tubiashii is included. The pathogenicity assays widen the host range of V. tubiashii to add two unreported species, Venerupis decussata and Donax trunculus, and the described as relatively resistant species V. philippinarum. PMID:25555343

  3. Morphological leaf variability in natural populations of Pistacia atlantica Desf. subsp. atlantica along climatic gradient: new features to update Pistacia atlantica subsp. atlantica key

    NASA Astrophysics Data System (ADS)

    El Zerey-Belaskri, Asma; Benhassaini, Hachemi

    2015-11-01

    The effect of bioclimate range on the variation in Pistacia atlantica Desf. subsp. atlantica leaf morphology was studied on 16 sites in Northwest Algeria. The study examined biometrically mature leaves totaling 3520 compound leaves. Fifteen characters (10 quantitative and 5 qualitative) were assessed on each leaf. For each quantitative character, the nested analysis of variance (ANOVA) was used to examine relative magnitude of variation at each level of the nested hierarchy. The correlation between the climatic parameters and the leaf morphology was examined. The statistical analysis applied on the quantitative leaf characters showed highly significant variation at the within-site level and between-site variation. The correlation coefficient (r) showed also an important correlation between climatic parameters and leaf morphology. The results of this study exhibited several values reported for the first time on the species, such as the length and the width of the leaf (reaching up to 24.5 cm/21.9 cm), the number of leaflets (up to 18 leaflets/leaf), and the petiole length of the terminal leaflet (reaching up to 3.4 cm). The original findings of this study are used to update the P. atlantica subsp. atlantica identification key.

  4. New triplex real-time PCR assay for detection of Mycobacterium avium subsp. paratuberculosis in bovine feces.

    PubMed

    Schönenbrücher, H; Abdulmawjood, A; Failing, K; Bülte, M

    2008-05-01

    In the present study, a robust TaqMan real-time PCR amplifying the F57 and the ISMav2 sequences of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples was developed and validated. The validation was based on the recommendations of International Organization for Standardization protocols for PCR and real-time PCR methods. For specificity testing, 205 bacterial strains were selected, including 105 M. avium subsp. paratuberculosis strains of bovine, ovine, and human origin and 100 non-M. avium subsp. paratuberculosis strains. Diagnostic quality assurance was obtained by use of an internal amplification control. By investigating six TaqMan reagents from different suppliers, the 100% detection probability was assessed to be 0.1 picogram M. avium subsp. paratuberculosis DNA per PCR. The amplification efficiency was 98.2% for the single-copy gene F57 and 97.8% for the three-copy insertion sequence ISMav2. The analytical method was not limited due to instrument specificity. The triplex real-time PCR allowed the reliable detection of M. avium subsp. paratuberculosis DNA using the ABI Prism 7000 sequence detection system, and the LightCycler 1.0. TaqMan(mgb) and locked nucleic acid fluorogenic probes were suitable for fluorescent signal detection. To improve the detection of M. avium subsp. paratuberculosis from bovine fecal samples, a more efficient DNA extraction method was developed, which offers the potential for automated sample processing. The 70% limit of detection was assessed to be 10(2) CFU per gram of spiked bovine feces. Comparative analysis of 108 naturally contaminated samples of unknown M. avium subsp. paratuberculosis status resulted in a relative accuracy of 98.9% and a sensitivity of 94.4% for fecal samples containing <10 CFU/g feces compared to the traditional culture method. PMID:18326682

  5. Mycobacterium avium subsp. paratuberculosis Antibody Response, Fecal Shedding, and Antibody Cross-Reactivity to Mycobacterium bovis in M. avium subsp. paratuberculosis-Infected Cattle Herds Vaccinated against Johne's Disease

    PubMed Central

    Hovingh, Ernest; Linscott, Rick; Martel, Edmond; Lawrence, John; Wolfgang, David; Griswold, David

    2014-01-01

    Vaccination for Johne's disease with killed inactivated vaccine in cattle herds has shown variable success. The vaccine delays the onset of disease but does not afford complete protection. Johne's disease vaccination has also been reported to interfere with measurements of cell-mediated immune responses for the detection of bovine tuberculosis. Temporal antibody responses and fecal shedding of Mycobacterium avium subsp. paratuberculosis, the causative agent of Johne's disease, were measured in 2 dairy cattle herds using Johne's disease vaccine (Mycopar) over a period of 7 years. Vaccination against Johne's disease resulted in positive serum M. avium subsp. paratuberculosis antibody responses in both herds, and the responses persisted in vaccinated cattle up to 7 years of age. Some vaccinated animals (29.4% in herd A and 36.2% in herd B) showed no serological reactivity to M. avium subsp. paratuberculosis. M. avium subsp. paratuberculosis-specific antibody responses were also detected in milk from Johne's disease-vaccinated animals, but fewer animals (39.3% in herd A and 49.4% in herd B) had positive results with milk than with serum samples. With vaccination against M. avium subsp. paratuberculosis, fecal shedding in both dairy herds was reduced significantly (P < 0.001). In addition, when selected Johne's disease-vaccinated and -infected animals were investigated for serological cross-reactivity to Mycobacterium bovis, no cross-reactivity was observed. PMID:24623626

  6. Novel Feature of Mycobacterium avium subsp. paratuberculosis, Highlighted by Characterization of the Heparin-Binding Hemagglutinin Adhesin

    PubMed Central

    Lefrancois, Louise H.; Bodier, Christelle C.; Cochard, Thierry; Canepa, Sylvie; Raze, Dominique; Lanotte, Philippe; Sevilla, Iker A.; Stevenson, Karen; Behr, Marcel A.; Locht, Camille

    2013-01-01

    Mycobacterium avium subsp. paratuberculosis comprises two genotypically defined groups, known as the cattle (C) and sheep (S) groups. Recent studies have reported phenotypic differences between M. avium subsp. paratuberculosis groups C and S, including growth rates, infectivity for macrophages, and iron metabolism. In this study, we investigated the genotypes and biological properties of the virulence factor heparin-binding hemagglutinin adhesin (HBHA) for both groups. In Mycobacterium tuberculosis, HBHA is a major adhesin involved in mycobacterium-host interactions and extrapulmonary dissemination of infection. To investigate HBHA in M. avium subsp. paratuberculosis, we studied hbhA polymorphisms by fragment analysis using the GeneMapper technology across a large collection of isolates genotyped by mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) and IS900 restriction fragment length polymorphism (RFLP-IS900) analyses. Furthermore, we analyzed the structure-function relationships of recombinant HBHA proteins of types C and S by heparin-Sepharose chromatography and surface plasmon resonance (SPR) analyses. In silico analysis revealed two forms of HBHA, corresponding to the prototype genomes for the C and S types of M. avium subsp. paratuberculosis. This observation was confirmed using GeneMapper on 85 M. avium subsp. paratuberculosis strains, including 67 strains of type C and 18 strains of type S. We found that HBHAs from all type C strains contain a short C-terminal domain, while those of type S present a long C-terminal domain, similar to that produced by Mycobacterium avium subsp. avium. The purification of recombinant HBHA from M. avium subsp. paratuberculosis of both types by heparin-Sepharose chromatography highlighted a correlation between their affinities for heparin and the lengths of their C-terminal domains, which was confirmed by SPR analysis. Thus, types C and S of M. avium subsp. paratuberculosis may be distinguished by the types of HBHA they produce, which differ in size and adherence properties, thereby providing new evidence that strengthens the genotypic differences between the C and S types of M. avium subsp. paratuberculosis. PMID:23974028

  7. Does reduced precipitation trigger physiological and morphological drought adaptations in European beech (Fagus sylvatica L.)? Comparing provenances across a precipitation gradient.

    PubMed

    Knutzen, Florian; Meier, Ina Christin; Leuschner, Christoph

    2015-09-01

    Global warming and associated decreases in summer rainfall may threaten tree vitality and forest productivity in many regions of the temperate zone in the future. One option for forestry to reduce the risk of failure is to plant genotypes which combine high productivity with drought tolerance. Growth experiments with provenances from different climates indicate that drought exposure can trigger adaptive drought responses in temperate trees, but it is not well known whether and to what extent regional precipitation reduction can increase the drought resistance of a species. We conducted a common garden growth experiment with five European beech (Fagus sylvatica L.) populations from a limited region with pronounced precipitation heterogeneity (816-544?mm?year(-1)), where phylogenetically related provenances grew under small to large water deficits. We grew saplings of the five provenances at four soil moisture levels (dry to moist) and measured ?30 morphological (leaf and root properties, root?:?shoot ratio), physiological (leaf water status parameters, leaf conductance) and growth-related traits (above- and belowground productivity) with the aim to examine provenance differences in the drought response of morphological and physiological traits and to relate the responsiveness to precipitation at origin. Physiological traits were more strongly influenced by provenance (one-third of the studied traits), while structural traits were primarily affected by water availability in the experiment (two-thirds of the traits). The modulus of leaf tissue elasticity ? reached much higher values late in summer in plants from moist origins resulting in more rapid turgor loss and a higher risk of hydraulic failure upon drought. While experimental water shortage affected the majority of morphological and productivity-related traits in the five provenances, most parameters related to leaf water status were insensitive to water shortage. Thus, plant morphology, and root growth in particular, did respond to reduced water availability with higher phenotypic plasticity than did physiology. We conclude that beech provenances exposed to different precipitation regimes have developed some genotypic differences with respect to leaf water status regulation, but these adaptations are associated with only minor adaptation in plant morphology and they do not affect the growth rate of the saplings. PMID:26209617

  8. Patterns of late spring frost leaf damage and recovery in a European beech (Fagus sylvatica L.) stand in south-eastern Germany based on repeated digital photographs

    NASA Astrophysics Data System (ADS)

    Menzel, Annette; Helm, Raimund; Zang, Christian

    2015-04-01

    The seasonality of woody plants in cold and temperate climates is adapted to the annual course of temperature and photoperiod in order to maximise the length of the active growing season and, at the same time, avoid damages by frost events, especially by late spring frosts. Winter chilling, spring warming and finally photoperiod trigger the timely bud burst of European beech (Fagus sylvatica L.) which as a climax species is quite sensitive to winter frost and also as seedling to late spring frosts. However, due to relatively late and less varying dates of leaf unfolding, damages by late spring frosts should not occur each year. In case of a total loss due to a late frost event, F. sylvatica trees produce a new set of leaves which guarantees survival, but diminishes carbon reserves. With a phenological camera we observed the phenological course of such an extreme event in the Nationalpark Bayerischer Wald in May 2011: Spring leaf unfolding, an almost complete loss of fresh green leaves after the frost event in the night 3rd to 4th May, a subsequent leafless period followed by re-sprouting. We modeled this special leaf development from day 80 to 210, observed as green% from the repeated digital camera pictures, using the Bayesian multiple change point approach recently introduced by Henneken et al. (2013). The results for more than 30 trees predominantly suggested a model with five change points: firstly, start of the season, abrupt ending before the frost event, the loss by the frost event and after a longer period of recovery the second leaf unfolding (St. John's sprout) ending in full leaf maturity. Analyzing the results of these models the following questions were answered (1) how long is the period of recovery till the second green-up? (2) does the temporal course of the second leafing differ from the first one? (3) what are the individual factors influencing damage and recovery? (4) are individuals with early or late bud burst more prone to damage? The five change points of the model almost perfectly matched the expected break points: i) start of the first greening between DOY (day of the year) 108 to 119 (mean 113), ii) end of greening and iii) visible frost damage after the frost on the night of May 3rd/4th (DOY 123/124), iv) re-sprouting 19 to 38 days after the frost, and v) full maturity around DOY 178 (166 to 184) when all beech crowns had fully recovered. Since frost damage was nearly 100%, individual susceptibility did not depend on the timing of first spring leaf unfolding. However, we could identify significant patterns in fitness linked to an earlier start of leaf unfolding.

  9. Impacts of repeated timber skidding on the chemical properties of topsoil, herbaceous cover and forest floor in an eastern beech (Fagus orientalis Lipsky) stand.

    PubMed

    Demir, Murat; Makineci, Ender; Comez, Aydin; Yilmaz, Ersel

    2010-07-01

    In this study, long-term timber skidding effects on herbaceous understory forest floor and soil were investigated on a skid road in a stand of the eastern beech (Fagus orientalis Lipsky). For this purpose, herbaceous understory forest floor and soil samples were collected from the skid road and from an undisturbed area used as a control plot. The mass (kg ha(-1)) of herbaceous and forest floor samples was determined, and soil characteristics were examined at two depths (0-5 cm and 5-10 cm). We quantified sand, silt and clay content, as well as bulk density compaction, pH, and organic carbon content in soil samples. The quantities of N, K, P, Na, Ca, Mg, Fe, Mn, Zn and Cu were determined in all herbaceous cover forest floor and soil samples. The quantities of Na, Fe, Zn, Cu and Mn in herbaceous understory samples from the skid road were considerably higher than those in the undisturbed area, while the quantity of Mg was considerably lower. These differences could have been caused by decreased herbaceous cover in addition to variations in the properties of the forest floor and soil after skidding. A lower amount of forest floor on the skid road was the result of skidding and harvesting activities. Mg and Zn contents in forest floor samples were found to be considerably lower for the skid road than for the undisturbed area. No significant differences were found in soil chemical properties (quantities of N, P, K, Na, Ca, Mg, Fe, Zn, Cu and Mn) at the 0-5 cm soil depth. Important differences exist between soil quantities of Mg at a 5-10 cm depth on the skid road and in undisturbed areas. Both 0-5 cm and 5-10 cm soil depths, the average penetrometer resistance values for the skid road was higher than for the undisturbed area. This result shows that the compaction caused by skidding is maintained to depth of 10 cm. Skid road soil showed higher bulk density values than undisturbed areas because of compaction. PMID:21186723

  10. Co-occurrence patterns of trees along macro-climatic gradients and their potential influence on the present and future distribution of Fagus sylvatica L.

    USGS Publications Warehouse

    Meier, E.S.; Edwards, T.C., Jr.; Kienast, Felix; Dobbertin, M.; Zimmermann, N.E.

    2011-01-01

    Aim During recent and future climate change, shifts in large-scale species ranges are expected due to the hypothesized major role of climatic factors in regulating species distributions. The stress-gradient hypothesis suggests that biotic interactions may act as major constraints on species distributions under more favourable growing conditions, while climatic constraints may dominate under unfavourable conditions. We tested this hypothesis for one focal tree species having three major competitors using broad-scale environmental data. We evaluated the variation of species co-occurrence patterns in climate space and estimated the influence of these patterns on the distribution of the focal species for current and projected future climates.Location Europe.Methods We used ICP Forest Level 1 data as well as climatic, topographic and edaphic variables. First, correlations between the relative abundance of European beech (Fagus sylvatica) and three major competitor species (Picea abies, Pinus sylvestris and Quercus robur) were analysed in environmental space, and then projected to geographic space. Second, a sensitivity analysis was performed using generalized additive models (GAM) to evaluate where and how much the predicted F. sylvatica distribution varied under current and future climates if potential competitor species were included or excluded. We evaluated if these areas coincide with current species co-occurrence patterns.Results Correlation analyses supported the stress-gradient hypothesis: towards favourable growing conditions of F. sylvatica, its abundance was strongly linked to the abundance of its competitors, while this link weakened towards unfavourable growing conditions, with stronger correlations in the south and at low elevations than in the north and at high elevations. The sensitivity analysis showed a potential spatial segregation of species with changing climate and a pronounced shift of zones where co-occurrence patterns may play a major role.Main conclusions Our results demonstrate the importance of species co-occurrence patterns for calibrating improved species distribution models for use in projections of climate effects. The correlation approach is able to localize European areas where inclusion of biotic predictors is effective. The climate-induced spatial segregation of the major tree species could have ecological and economic consequences. ?? 2010 Blackwell Publishing Ltd.

  11. The effect of carbohydrate accumulation and nitrogen deficiency on feedback regulation of photosynthesis in beech (Fagus sylvatica) under elevated CO2 concentration

    NASA Astrophysics Data System (ADS)

    Klem, K.; Urban, O.; Holub, P.; Rajsnerova, P.

    2012-04-01

    One of the main manifestations of global change is an increase in atmospheric CO2 concentration. Elevated concentration of CO2 has stimulating effect on plant photosynthesis and consequently also on the productivity. Long-term studies, however, show that this effect is progressively reduced due to feedback regulation of photosynthesis. The main causes of this phenomenon are considered as two factors: i) increased biomass production consumes a larger amount of nitrogen from the soil and this leads to progressive nitrogen limitation of photosynthesis, particularly at the level of the enzyme Rubisco, ii) the sink capacity is genetically limited and elevated CO2 concentration leads to increased accumulation of carbohydtrates (mainly sucrose, which is the main transport form of assimilates) in leaves. Increased concentrations of carbohydrates leads to a feedback regulation of photosynthesis by both, long-term feedback regulation of synthesis of the enzyme Rubisco, and also due to reduced capacity to produce ATP in the chloroplasts. However, mechanisms for interactive effects of nitrogen and accumulation of non-structural carbohydrates are still not well understood. Using 3-year-old Fagus sylvatica seedlings we have explored the interactive effects of nitrogen nutrition and sink capacity manipulation (sucrose feeding) on the dynamics of accumulation of non-structural carbohydrates and changes in photosynthetic parameters under ambient (385 ?mol (CO2) mol-1) and elevated (700 ?mol(CO2) mol-1) CO2 concentration. Sink manipulation by sucrose feeding led to a continuous increase of non-structural carbohydrates in leaves, which was higher in nitrogen fertilized seedlings. The accumulation of non-structural carbohydrates was also slightly stimulated by elevated CO2 concentration. Exponential decay (p <0.01) was observed in CO2 assimilation rate and stomatal conductance when the content of non-structural carbohydrates increased. However, this relationship was modified by the nitrogen content. Accumulation of non-structural carbohydrates had relatively smaller effect on actual quantum yield of photosystem II. Both, CO2 assimilation rate and the actual quantum yield of photosystem II decreased more rapidly during sink manipulation in elevated concentrations of CO2 than in ambient. Application of chlorophyll fluorescence imaging enabled us to evaluate changes in spatial distribution of feedback regulation of photosynthesis on the leaf-level. We can conclude that the accumulation of non-structural carbohydrates down-regulates photosynthesis mainly through the stomatal conductance, and this effect is further modified by nitrogen content.

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

    PubMed

    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. PMID:9160324

  13. Silencing of host basal defense response-related gene expression increases susceptibility of Nicotiana benthamiana to Clavibacter michiganensis subsp. michiganensis.

    PubMed

    Balaji, Vasudevan; Sessa, Guido; Smart, Christine D

    2011-03-01

    Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host-pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana-C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato. PMID:21062112

  14. Prevalence and Acquisition of the Genes for Zoocin A and Zoocin A Resistance in Streptococcus equi subsp. zooepidemicus

    PubMed Central

    O’Rourke, Anna-Lee D.; Simmonds, Robin S.; Gargis, Amy S.; Sloan, Gary L.

    2009-01-01

    Zoocin A is a streptococcolytic enzyme produced by Streptococcus equi subsp. zooepidemicus strain 4881. The zoocin A gene (zooA) and the gene specifying resistance to zoocin A (zif) are adjacent on the chromosome and are divergently transcribed. Twenty-four S. equi subsp. zooepidemicus strains were analyzed to determine the genetic difference between three previously characterized as zoocin A producers (strains 4881, 9g and 9h) and the twenty-one non-producers. LT-PCR and Southern hybridization studies revealed that none of the non-producer strains possessed zooA or zif. RAPD and PFGE showed that the 24 strains were a genetically diverse population with 8 RAPD profiles. S. equi subsp. zooepidemicus strains 9g and 9h appeared genetically identical to each other, but quite different from 4881. Sequences derived from 4881 and 9g showed that zooA and zif were integrated into the chromosome adjacent to the gene flaR. A comparison of these sequences with the genome sequences of S. equi subsp. zooepidemicus strains H70 and MGCS10565 and S. equi subsp. equi strain 4047 suggests that flaR flanks a region of genome plasticity in this species. PMID:19357799

  15. Prevalence and acquisition of the genes for zoocin A and zoocin A resistance in Streptococcus equi subsp. zooepidemicus.

    PubMed

    Gargis, Amy S; O'Rourke, Anna-Lee D; Sloan, Gary L; Simmonds, Robin S

    2009-05-01

    Zoocin A is a streptococcolytic enzyme produced by Streptococcus equi subsp. zooepidemicus strain 4881. The zoocin A gene (zooA) and the gene specifying resistance to zoocin A (zif) are adjacent on the chromosome and are divergently transcribed. Twenty-four S. equi subsp. zooepidemicus strains were analyzed to determine the genetic difference among three previously characterized as zoocin A producers (strains 4881, 9g, and 9h) and the 21 nonproducers. LT-PCR and Southern hybridization studies revealed that none of the nonproducer strains possessed zooA or zif. RAPD and PFGE showed that the 24 strains were a genetically diverse population with eight RAPD profiles. S. equi subsp. zooepidemicus strains 9g and 9h appeared to be genetically identical to each other but quite different from strain 4881. Sequences derived from 4881 and 9g showed that zooA and zif were integrated into the chromosome adjacent to the gene flaR. A comparison of these sequences with the genome sequences of S. equi subsp. zooepidemicus strains H70 and MGCS10565 and S. equi subsp. equi strain 4047 suggests that flaR flanks a region of genome plasticity in this species. PMID:19357799

  16. Complete Genome Sequence of Xanthomonas citri subsp. citri Strain Aw12879, a Restricted-Host-Range Citrus Canker-Causing Bacterium

    PubMed Central

    Jalan, Neha; Kumar, Dibyendu; Yu, Fahong; Jones, Jeffrey B.; Graham, James H.

    2013-01-01

    Xanthomonas citri subsp. citri causes citrus canker. The Asiatic strain has a broad host range, whereas the Wellington variant has a restricted host range. Here, we present the complete genome of X. citri subsp. citri strain AW12879. This study lays the foundation to further characterize the mechanisms for virulence and host range of X. citri. PMID:23682143

  17. Modulation of Cytokine Gene Expression and Secretion During the Periparturient Period in Dairy Cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Modulation of cytokine gene expression and secretion during the periparturient period in dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis Technical abstract Johne’s disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is estimated to infect more t...

  18. Highly Specific and Quick Detection of Mycobacterium avium subsp. paratuberculosis in Feces and Gut Tissue of Cattle and Humans by Multiple Real-Time PCR Assays?

    PubMed Central

    Imirzalioglu, Can; Dahmen, Heinrich; Hain, Torsten; Billion, Andre; Kuenne, Carsten; Chakraborty, Trinad; Domann, Eugen

    2011-01-01

    Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease (JD) in cattle and may be associated with Crohn's disease (CD) in humans. It is the slowest growing of the cultivable mycobacteria, and culture from clinical, veterinary, food, or environmental specimens can take 4 months or even longer. Currently, the insertion element IS900 is used to detect M. avium subsp. paratuberculosis DNA. However, closely related IS900 elements are also present in other mycobacteria, thus limiting its specificity as a target. Here we describe the use of novel primer sets derived from the sequences of two highly specific single copy genes, MAP2765c and MAP0865, for the quantitative detection of M. avium subsp. paratuberculosis within 6 h by using real-time PCR. Specificity of the target was established using 40 M. avium subsp. paratuberculosis isolates, 67 different bacterial species, and two intestinal parasites. Using the probes and methods described, we detected 27 (2.09%) M. avium subsp. paratuberculosis-positive stool specimens from 1,293 individual stool samples by the use of either IS900 or probes deriving from the MAP2765c and MAP0865 genes described here. In general, bacterial load due to M. avium subsp. paratuberculosis was uniformly low in these samples and we estimated 500 to 5,000 M. avium subsp. paratuberculosis bacteria per gram of stool in assay-positive samples. Thus, the methods described here are useful for rapid and specific detection of M. avium subsp. paratuberculosis in clinical samples. PMID:21430100

  19. Nineteen Whole-Genome Assemblies of Yersinia pestis subsp. microtus, Including Representatives of Biovars caucasica, talassica, hissarica, altaica, xilingolensis, and ulegeica

    PubMed Central

    Bogun, Aleksandr G.; Kadnikova, Lidiya A.; Maiskaya, Nadezhda V.; Platonov, Mikhail E.; Anisimov, Nikolai V.; Galkina, Elena V.; Dentovskaya, Svetlana V.

    2015-01-01

    The etiologic agent of plague, Yersinia pestis, includes two subspecies, of which Y. pestis subsp. microtus contains the strains that cause only occasional diseases in humans that are not accompanied by human-to-human transmission. Here, we report the draft genome sequences of 19 Y. pestis strains (across 6 biovars of Y. pestis subsp. microtus). PMID:26634751

  20. Nineteen Whole-Genome Assemblies of Yersinia pestis subsp. microtus, Including Representatives of Biovars caucasica, talassica, hissarica, altaica, xilingolensis, and ulegeica.

    PubMed

    Kislichkina, Angelina A; Bogun, Aleksandr G; Kadnikova, Lidiya A; Maiskaya, Nadezhda V; Platonov, Mikhail E; Anisimov, Nikolai V; Galkina, Elena V; Dentovskaya, Svetlana V; Anisimov, Andrey P

    2015-01-01

    The etiologic agent of plague, Yersinia pestis, includes two subspecies, of which Y. pestis subsp. microtus contains the strains that cause only occasional diseases in humans that are not accompanied by human-to-human transmission. Here, we report the draft genome sequences of 19 Y. pestis strains (across 6 biovars of Y. pestis subsp. microtus). PMID:26634751

  1. GENERAL TECHNICAL REPORT PSW-GTR-240 Screening for Resistance to Beech Bark Disease

    E-print Network

    American beech (Fagus grandifolia Ehrh.) trees since the accidental introduction of the beech scale insect scale insect (Cryptococcus fagisuga) is the insect component, and a canker-causing Neonectria (either and David W. Carey1 Abstract Beech bark disease (BBD) is an insect-disease complex that has been killing

  2. The first host records for the Nearctic species Triraphis discoideus (Hymenoptera: Braconidae: Rogadinae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Limacodid larvae were collected from 2004 – 2007 on leaves of the following host plants in the District of Columbia and Maryland: Carya glabra, pignut hickory; Quercus alba, white oak; Quercus rubra, northern red oak; Nyssa sylvatica, black gum; Prunus serotina, black cherry; and Fagus grandifolia, ...

  3. Development of an F57 sequence-based real-time PCR assay for detection of Mycobacterium avium subsp. paratuberculosis in milk.

    PubMed

    Tasara, T; Stephan, R

    2005-10-01

    A light cycler-based real-time PCR (LC-PCR) assay that amplifies the F57 sequence of Mycobacterium avium subsp. paratuberculosis was developed. This assay also includes an internal amplification control template to monitor the amplification conditions in each reaction. The targeted F57 sequence element is unique for M.avium subsp. paratuberculosis and is not known to exist in any other bacterial species. The assay specificity was demonstrated by evaluation of 10 known M. avium subsp. paratuberculosis isolates and 33 other bacterial strains. The LC-PCR assay has a broad linear range (2 x 10(1) to 2 x10(6) copies) for quantitative estimation of the number of M. avium subsp. paratuberculosis F57 target copies in positive samples. To maximize the assay's detection sensitivity, an efficient strategy for isolation of M. avium subsp. paratuberculosis DNA from spiked milk samples was also developed. The integrated procedure combining optimal M. avium subsp. paratuberculosis DNA isolation and real-time PCR detection had a reproducible detection limit of about 10 M. avium subsp. paratuberculosis cells per ml when a starting sample volume of 10 ml of M. avium subsp. paratuberculosis-spiked milk was analyzed. The entire process can be completed within a single working day and is suitable for routine monitoring of milk samples for M. avium subsp. paratuberculosis contamination. The applicability of this protocol for naturally contaminated milk was also demonstrated using milk samples from symptomatic M. avium subsp. paratuberculosis-infected cows, as well as pooled samples from a dairy herd with a confirmed history of paratuberculosis. PMID:16204510

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

    PubMed

    Lund, Marie B; Schätzle, Sarah; Schramm, Andreas; Kjeldsen, Kasper U

    2012-03-01

    Clone library-based studies have shown that almost all lumbricid earthworm species harbour host-specific symbiotic bacteria belonging to the novel genus Verminephrobacter in their nephridia (excretory organs). To date the only described representative from this genus is Verminephrobacter eiseniae, the specific symbiont of the earthworm Eisenia fetida. In this study two novel rod-shaped, non-endosporeforming, betaproteobacterial symbionts were isolated from the nephridia of two closely related earthworm species. Both isolates were affiliated with the genus Verminephrobacter by 16S rRNA gene sequence 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 environment. Based on 16S rRNA gene similarity, DNA-DNA hybridization value and phenotypic characteristics the two isolates are clearly distinct from V. eiseniae. Phenotypic characteristics could not clearly differentiate the two strains as separate species but a low DNA-DNA hybridization value of 57.3%, 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, At4(T) = DSM 21361(T) = LMG 25313(T)) and V. aporrectodeae subsp. caliginosae (type strain, Ac9(T) = DSM 21895(T) = LMG 25312(T)) isolated from the nephridia of the earthworms Aporrectodea tuberculata and A. caliginosa, respectively. PMID:22041977

  5. Characterization of the gene for an immunodominant 72 kDa lipoprotein of Mycoplasma mycoides subsp. mycoides small colony type.

    PubMed

    Cheng, X; Nicolet, J; Miserez, R; Kuhnert, P; Krampe, M; Pilloud, T; Abdo, E M; Griot, C; Frey, J

    1996-12-01

    With the aim of characterizing specific immunogenic proteins of Mycoplasma mycoides subsp. mycoides small colony (SC) type, the aetiological agent of contagious bovine pleuropneumonia, a gene encoding a major immunogenic protein of 72 kDa named P72 was cloned and expressed in Escherichia coli. The expressed protein was of the same apparent molecular mass as that produced by the parent strain. The predicted molecular mass of P72, based on the DNA-deduced amino acid sequence, was 61.118 kDa, significantly lower than the apparent molecular mass of endogenous or recombinant P72 on SDS-PAGE. Analysis of the amino acid sequence revealed a typical prokaryotic signal peptidase II-membrane lipoprotein lipid attachment site and a transmembrane structure domain in the leader sequence at the amino-terminal end of the protein. P72 was shown to be a lipoprotein and its surface location was confirmed by trypsin treatment of whole cells. An unassigned gene encoding a peptide with some similarity to P72 was found on the genome sequence of M. capricolum subsp. capricolum but not on that of Mycoplasma genitalium. The P72 gene was detected in 11/11 M. mycoides subsp. mycoides SC strains. Antiserum against recombinant P72 reacted strongly with 12/12 strains of M. mycoides subsp. mycoides SC, weakly with Mycoplasma bovine group 7 strain PG50, but not with other members of the 'mycoides cluster' or closely related mycoplasmas. Cows experimentally contact-infected with M. mycoides subsp. mycoides SC developed a humoral response against P72 within 35 d. P72 is a specific antigenic membrane lipoprotein of M. mycoides subsp. mycoides SC with potential for use in development of diagnostic reagents. It seems to belong to a family of lipoproteins of the "mycoides cluster'. PMID:9004514

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

    PubMed Central

    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

  7. Phytochemical Composition and Antinociceptive Activity of Bauhinia glauca subsp. hupehana in Rats

    PubMed Central

    Xu, Jinlong; Zhao, Qizhi; Wei, Lei; Yang, Yu; Xu, Rui; Yu, Nengjiang; Zhao, Yimin

    2015-01-01

    In traditional medicine, Bauhinia glauca subsp. hupehana has long been used as an analgesic agent in China. The aim of this study was to evaluate the antinociceptive activity of the ethanol extract of the aerial parts of B. glauca subsp. hupehana (BHE) in rats and its chemical fingerprint. The antinociceptive activity of BHE was assessed in mice using chemically and heat–induced pain models, such as the acetic acid–induced writhing, hot plate, tail–flick and glutamate tests. Naltrexone hydrochloride, a non–selective opioid receptor antagonist, was utilized to determine the involvement of the opioid system. In addition to this, the involvements of the cGMP and ATP–sensitive K+ channel pathways were also detected using methylene blue and glibenclamide. The oral administration of BHE (at doses of 50, 100 and 200 mg/kg) produced significant and dose–related inhibitions in both the chemically and heat–induced pain models. Interestingly, in the abdominal constriction test, when the dose of BHE was increased to 800 mg/kg (p.o., n = 10), the inhibition rate was 100%. The antinociceptive mechanism may involve the cGMP pathway and ATP sensitive K+ channel pathway. The central antinociceptive effect was not antagonized by naltrexone. One phenolic acid, one lignin and five flavonoids were isolated from BHE. The antinociceptive activity of BHE was most likely due to the presence of the flavonoids. The acute toxicity results showed that BHE was safe at a high dose (2 g/kg, p.o.). The current investigation demonstrates that B. glauca subsp. hupehana is a potential candidate for the development of novel, non–opioid, analgesic phytomedicines. PMID:25658740

  8. DrsG from Streptococcus dysgalactiae subsp. equisimilis inhibits the antimicrobial peptide LL-37.

    PubMed

    Smyth, Danielle; Cameron, Ainslie; Davies, Mark R; McNeilly, Celia; Hafner, Louise; Sriprakash, Kadaba S; McMillan, David J

    2014-06-01

    SIC and DRS are related proteins present in only 4 of the >200 Streptococcus pyogenes emm types. These proteins inhibit complement-mediated lysis and/or the activity of certain antimicrobial peptides (AMPs). A gene encoding a homologue of these proteins, herein called DrsG, has been identified in the related bacterium Streptococcus dysgalactiae subsp. equisimilis. Here we show that geographically dispersed isolates representing 14 of 50 emm types examined possess variants of drsG. However, not all isolates within the drsG-positive emm types possess the gene. Sequence comparisons also revealed a high degree of conservation in different S. dysgalactiae subsp. equisimilis emm types. To examine the biological activity of DrsG, recombinant versions of two major DrsG variants, DrsGS and DrsGL, were expressed and purified. Western blot analysis using antisera raised to these proteins demonstrated both variants to be expressed and secreted into culture supernatants. Unlike SIC, but similar to DRS, DrsG does not inhibit complement-mediated lysis. However, like both SIC and DRS, DrsG is a ligand of the cathelicidin LL-37 and is inhibitory to its bactericidal activity in in vitro assays. Conservation of prolines in the C-terminal region also suggests that these residues are important in the biology of this family of proteins. This is the first report demonstrating the activity of an AMP-inhibitory protein in S. dysgalactiae subsp. equisimilis and suggests that inhibition of AMP activity is the primary function of this family of proteins. The acquisition of the complement-inhibitory activity of SIC may reflect its continuing evolution. PMID:24664506

  9. Phytochemical composition and antinociceptive activity of Bauhinia glauca subsp. hupehana in rats.

    PubMed

    Xu, Jinlong; Zhao, Qizhi; Wei, Lei; Yang, Yu; Xu, Rui; Yu, Nengjiang; Zhao, Yimin

    2015-01-01

    In traditional medicine, Bauhinia glauca subsp. hupehana has long been used as an analgesic agent in China. The aim of this study was to evaluate the antinociceptive activity of the ethanol extract of the aerial parts of B. glauca subsp. hupehana (BHE) in rats and its chemical fingerprint. The antinociceptive activity of BHE was assessed in mice using chemically and heat-induced pain models, such as the acetic acid-induced writhing, hot plate, tail-flick and glutamate tests. Naltrexone hydrochloride, a non-selective opioid receptor antagonist, was utilized to determine the involvement of the opioid system. In addition to this, the involvements of the cGMP and ATP-sensitive K+ channel pathways were also detected using methylene blue and glibenclamide. The oral administration of BHE (at doses of 50, 100 and 200 mg/kg) produced significant and dose-related inhibitions in both the chemically and heat-induced pain models. Interestingly, in the abdominal constriction test, when the dose of BHE was increased to 800 mg/kg (p.o., n = 10), the inhibition rate was 100%. The antinociceptive mechanism may involve the cGMP pathway and ATP sensitive K+ channel pathway. The central antinociceptive effect was not antagonized by naltrexone. One phenolic acid, one lignin and five flavonoids were isolated from BHE. The antinociceptive activity of BHE was most likely due to the presence of the flavonoids. The acute toxicity results showed that BHE was safe at a high dose (2 g/kg, p.o.). The current investigation demonstrates that B. glauca subsp. hupehana is a potential candidate for the development of novel, non-opioid, analgesic phytomedicines. PMID:25658740

  10. High-Throughput Direct Fecal PCR Assay for Detection of Mycobacterium avium subsp. paratuberculosis in Sheep and Cattle

    PubMed Central

    Waldron, Anna M.; Galea, Francesca; Whittington, Ann-Michele; Saunders, Vanessa F.; Begg, Douglas J.; de Silva, Kumudika; Purdie, Auriol C.; Whittington, Richard J.

    2014-01-01

    Johne's disease (JD) is a chronic enteric disease caused by Mycobacterium avium subsp. paratuberculosis that affects ruminants. Transmission occurs by the fecal-oral route. A commonly used antemortem diagnostic test for the detection of M. avium subsp. paratuberculosis in feces is liquid culture; however, a major constraint is the 2- to 3-month incubation period needed for this method. Rapid methods for the detection of M. avium subsp. paratuberculosis based on PCR have been reported, but comprehensive validation data are lacking. We describe here a new test, the high-throughput-Johnes (HT-J), to detect M. avium subsp. paratuberculosis in feces. Its diagnostic accuracy was compared with that of liquid radiometric (Bactec) fecal culture using samples from cattle (1,330 samples from 23 herds) and sheep (596 samples from 16 flocks). The multistage protocol involves the recovery of M. avium subsp. paratuberculosis cells from a fecal suspension, cell rupture by bead beating, extraction of DNA using magnetic beads, and IS900 quantitative PCR. The limit of detection of the assay was 0.0005 pg, and the limit of quantification was 0.005 pg M. avium subsp. paratuberculosis genomic DNA. Only M. avium subsp. paratuberculosis was detected from a panel of 51 mycobacterial isolates, including 10 with IS900-like sequences. Of the 549 culture-negative fecal samples from unexposed herds and flocks, 99% were negative in the HT-J test, while 60% of the bovine- and 84% of the ovine-culture-positive samples were positive in the HT-J test. As similar total numbers of samples from M. avium subsp. paratuberculosis-exposed animals were positive in culture and HT-J tests in both species, and as the results of a McNemar's test were not significant, these methods probably have similar sensitivities, but the true diagnostic sensitivities of these tests are unknown. These validation data meet the consensus-based reporting standards for diagnostic test accuracy studies for paratuberculosis and the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines (S. A. Bustin et al., Clin. Chem. 55:611–622, 2009, doi:10.1373/clinchem.2008.112797). The HT-J assay has been approved for use in JD control programs in Australia and New Zealand. PMID:24352996

  11. A UV tolerant mutant of Bacillus thuringiensis subsp. kurstaki producing melanin.

    PubMed

    Saxena, Deepak; Ben-Dov, Eitan; Manasherob, Robert; Barak, Ze'ev; Boussiba, Sammy; Zaritsky, Arieh

    2002-01-01

    A UV-resistant mutant (Bt-m) of Bacillus thuringiensis subsp. kurstaki, producing a dark brown pigment, identified as melanin, was studied. Bt-m had higher larvicidity against Heliothis armigera than its parent. Survival of Bt-m spores and their insecticidal activity to irradiation at 254 nm and 366 nm were higher than those of the parent. The only toxic polypeptide produced by Bt-m was Cry1Ac (130 kDa); it lost cry1Aa, cry2Aa, and cry2Ab. PMID:11727037

  12. Branched-chain amino acid biosynthesis genes in Lactococcus lactis subsp. lactis.

    PubMed Central

    Godon, J J; Chopin, M C; Ehrlich, S D

    1992-01-01

    The genes for biosynthesis of the branched-chain amino acids leucine, isoleucine, and valine in Lactococcus lactis subsp. lactis NCDO2118 were characterized by cloning, complementation in Escherichia coli and Bacillus subtilis, and nucleotide sequence analysis. Nine structural genes are clustered on a 12-kb DNA fragment in the order leuABCD ilvDBNCA. Upstream of these genes, the nucleotide sequence suggests the existence of regulation by transcriptional attenuation. Between the leuD and ilvD genes is an unexpected gene, encoding a protein which belongs to the ATP-binding cassette protein superfamily. PMID:1400210

  13. Genetic IS901 RFLP diversity among Mycobacterium avium subsp. avium isolates from four pheasant flocks.

    PubMed

    Moravkova, Monika; Lamka, Jiri; Slany, Michal; Pavlik, Ivo

    2013-01-01

    IS901 RFLP analysis of 36 Mycobacterium avium subsp. avium (MAA) isolates from 15 pheasants (Phasianus colchicus) and two goshawks (Accipiter gentilis) from four pheasant farms was performed. Using this method, six different IS901 RFLP types (E, F, G, M, Q, and V) were identified. The distribution of IS901 RFLP profiles was tightly linked to individual flocks. Matching IS901 RFLP profiles observed in the present study indicate MAA transmission between pheasants and goshawks in the same locality. In two flocks, different pheasants within a flock as well as in various organs of five individual pheasants were found to have two distinct IS901 RFLP profiles. PMID:23388436

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

    PubMed Central

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

    2012-01-01

    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. PMID:23242372

  15. A circular genetic map of Erwinia carotovora subsp. atroseptica 3-2

    SciTech Connect

    Nikolaichik, E.A.; Pesnyakevich, A.G.

    1995-08-01

    A circular genetic map of Erwinia carotovora subsp. atroseptica 3-2 was constructed on the basis of the R471a plasmid and Tn5 and Tn9 using Hfr-like donors. Forty-six genes, including phytopathogenicity genes, were located on the basis of interrupted mating experiment results and analysis of coinheritance of markers on a map of 183 min in length. The similarity and differences of chromosomal genetic maps of Erwinia genus bacteria are discussed. 23 refs., 2 figs., 4 tabs.

  16. Characterization of Blood Culture Isolates of Streptococcus dysgalactiae subsp. equisimilis Possessing Lancefield's Group A Antigen

    PubMed Central

    Brandt, Claudia M.; Haase, Gerhard; Schnitzler, Norbert; Zbinden, Reinhard; Lütticken, Rudolf

    1999-01-01

    For three human blood culture isolates of beta-hemolytic streptococci with Lancefield's serogroup A antigen, phylogenetic analysis of the 16S rRNA genes confirmed biochemical identification as Streptococcus dysgalactiae subsp. equisimilis. Genes encoding M or M-like proteins, which are considered to be major virulence determinants in streptococci, were detected in all of these strains. Our data clearly demonstrate that for beta-hemolytic streptococci, the species assignment should not be based on the results of serogrouping alone. PMID:10565964

  17. Carbon assimilation, translocation and respiration in Fagus sylvatica and Abies alba stands measured by gas exchange and isotopic techniques during two contrasting climatic years

    NASA Astrophysics Data System (ADS)

    Gavrichkova, Olga; Scartazza, Andrea; Zampedri, Roberto; Cavagna, Mauro; Sottocornola, Matteo; Matteucci, Giorgio; Brugnoli, Enrico

    2014-05-01

    Global warming is tremendously influencing the climate of mountain areas through constantly rising temperatures and changes in local hydrological cycle. Increase of precipitation extremes, seasonal shifts of rainfall regime, heat waves are becoming more and more frequent events here. Vulnerability and plasticity of the local individual tree species under changing climate has still to be evaluated under field conditions. Two consecutive years, 2012 and 2013 were quite distinct in the climatic conditions during the plant growing season. Summer 2012 was characterized by a prolonged summer drought with almost no precipitation in central Italy from the end of May up to the end of August. The situation was aggravated by a very dry winter during this year. Mean annual temperatures in 2012 were 2oC higher in respect to the temperatures measured in the last 10 years. Conversely, year 2013 was milder with occasional rain events also during the summer months and temperatures close to the average values. In the Alpine zone the difference between two years were less pronounced with 2012 being slightly warmer than average and 2013 was characterized by unusually abundant spring precipitations. Taking advantage of these two contrasting years, we have monitored a functional response of one deciduous and one coniferous mountain forest stands growing in different mountain climate zones to variations in the local climate. The first, a deciduous European beech (Fagus sylvatica) forest, is located in the Appennine region of Italy at 1700 m height (Collelongo site, AQ) and characterized by a Mountain-Mediterranean climate. The second is a mixed forest dominated by Silver fir (Abies alba) which was chosen as a target species for our study. The site is located at 1350m height in the south-eastern Alps (Lavarone, TN) and is characterized by a mountain temperate climate. Sampling of plant material and point flux measurements were performed in the beginning, middle and the end of the growing season each year. At the beech site the middle samplings corresponded to the peak of the drought season whereas the last samplings of each year - to recovery phase. Leaves were sampled with three replicates at three heights. Assimilation activity was monitored on the leaf level with a portable LiCor 6400 system. Leaf respiration was measured with the same instrument after keeping the leaves for 30 min in darkness. Recently assimilated soluble sugars as well as bulk leaf organic matter were analysed in the laboratory for their ?13C signature and for sugars quantity and composition. Trunk, root and soil respirations together with their ?13C signatures were measured with closed static chambers by the Keeling plot approach. Phloem was sampled with a bark core aiming to analyse the C isotopic signature and composition of assimilates translocate downward with the phloem flow. A sequence of climatically different growing seasons and detailed analyses of plant material allowed us to evaluate climatically-induced variations in different steps of the C cycle at a plant level and to derive some conclusions on the plasticity of European beech and Silver fir in response to changing climate.

  18. Persistence of Mycobacterium avium subsp. paratuberculosis at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle.

    PubMed

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

    2011-05-01

    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 (10(1) 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 (10(2) 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

  19. Multi-Method Approach for Characterizing the Interaction between Fusarium verticillioides and Bacillus thuringiensis Subsp. Kurstaki

    PubMed Central

    Rocha, Liliana O.; Tralamazza, Sabina Moser.; Reis, Gabriela M.; Rabinovitch, Leon; Barbosa, Cynara B.; Corrêa, Benedito

    2014-01-01

    Bacterial antagonists used as biocontrol agents represent part of an integrated management program to reduce pesticides in the environment. Bacillus thuringiensis is considered a good alternative as a biocontrol agent for suppressing plant pathogens such as Fusarium. In this study, we used microscopy, flow cytometry, indirect immunofluorescence, and high performance liquid chromatography to determine the interaction between B. thuringiensis subsp. kurstaki LFB-FIOCRUZ (CCGB) 257 and F. verticillioides MRC 826, an important plant pathogen frequently associated with maize. B. thuringiensis showed a strong in vitro suppressive effect on F. verticillioides growth and inhibited fumonisin production. Flow cytometry analysis was found to be adequate for characterizing the fungal cell oscillations and death during these interactions. Further studies of the antagonistic effect of this isolate against other fungi and in vivo testing are necessary to determine the efficacy of B. thuringiensis subsp. kurstaki in controlling plant pathogens. This is the first report on the use of flow cytometry for quantifying living and apoptotic F. verticillioides cells and the B. thuringiensis Cry 1Ab toxin. PMID:24739804

  20. First time isolation of Photobacterium damselae subsp. damselae from Caranx sexfasciatus in Persian Gulf, Iran

    PubMed Central

    Hassanzadeh, Yashgin; Bahador, Nima; Baseri-Salehi, Majid

    2015-01-01

    Background and Objective: Photobacterium damselae subsp. damselae is a marine pathogenic bacterium which causes disease in marine animals and human. This bacterium mostly found in coastal shallow seawater. So, the aim of this study was isolation and characterization of Photobacterium damselae subsp. damselae from edible fish of Persian Gulf, Bandar Abbas. Material and Methods: Totally 100 fish from different species were evaluated and out of that 5 different types of fish with external symptoms including: Caranx sexfasciatus, Lethrinus olivaceus, Scomberoid tol, Auxis thazard and Liza macrolepis, were collected from Bandar Abbas local fish market in September 2013. The samples were cultured on Marin Agar 2216 and Thiosulfate Citrate Bile salts Sucrose Agar media and incubated at 25°C for 48 hrs. Then the isolates were characterized using biochemical (API 20 NE system) and molecular techniques. In addition, antibiotic susceptibility, presence of poly ? hydroxy butyrate and hemolysis activity of isolates were evaluated. Results and Conclusion: Entirely, 30 Gram negative bacterial colonies were isolated from the selected fish. Among the isolates, two suspected colonies were identified as Photobacterium damselae from Caranx sexfasciatus with API 20NE biochemical test. This results confirmed by 16s rRNA sequencing method. Both isolates showed ? hemolytic with existence of ? hydroxyl butyrate. Furthermore, the isolates were susceptible to ciprofloxacin, chloramphenicol and nalidixic acid. Conclusion: Overall, the study indicated first time isolation of this bacterium from one type of fish caught from Persian Gulf, which warns us to pay more attention to fishery in this geographical area. PMID:26668707

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

    PubMed Central

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

    2015-01-01

    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.

  2. Cyclopropane fatty acid synthase from Oenococcus oeni: expression in Lactococcus lactis subsp. cremoris and biochemical characterization.

    PubMed

    To, Thi Mai Huong; Grandvalet, Cosette; Alexandre, Hervé; Tourdot-Maréchal, Raphaëlle

    2015-11-01

    Bacterial cyclopropane fatty acid synthases (CFA synthases) catalyze the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to the double bond of a lipid chain, thereby forming a cyclopropane ring. CFAs contribute to resistance to acidity, dryness, and osmotic imbalance in many bacteria. This work describes the first biochemical characterization of a lactic acid bacterium CFA synthase. We have overexpressed Oenococcus oeni CFA synthase in E. coli in order to purify the enzyme. The optimum cyclopropanation activity was obtained at pH 5.6 and 35.8 °C. The high K m (AdoMet) value obtained (2.26 mM) demonstrates the low affinity of O. oeni enzyme toward the L. lactis subsp. cremoris unsaturated phospholipids. These results explain the partial complementation of the L. lactis subsp. cremoris cfa mutant by the O. oeni cfa gene and suggest a probable substrate specificity of the O. oeni enzyme. The current study reveals an essential hypothesis about the specificity of O. oeni CFA synthase which could play a key function in the acid tolerance mechanisms of this enological bacterium. PMID:26294376

  3. Detection of Mycobacterium avium subsp. paratuberculosis in infected tissues by new species-specific immunohistological procedures.

    PubMed

    Coetsier, C; Havaux, X; Mattelard, F; Sadatte, S; Cormont, F; Buergelt, K; Limbourg, B; Latinne, D; Bazin, H; Denef, J F; Cocito, C

    1998-07-01

    We have previously described the cloning and sequencing of a gene portion coding for the terminal part of a 34-kDa protein of Mycobacterium avium subsp. paratuberculosis, the etiological agent of Johne's disease (P. Gilot, M. De Kesel, L. Machtelinckx, M. Coene, and C. Cocito, J. Bacteriol. 175:4930-4935, 1993). The recombinant polypeptide (a362) carries species-specific B-cell epitopes which do not cross-react with other mycobacterial pathogens (M. De Kesel, P. Gilot, M.-C. Misonne, M. Coene, and C. Cocito, J. Clin. Microbiol. 31:947-954, 1993). The present work describes the preparation of polyclonal and monoclonal antibodies directed against a362 and the use of these immunoglobulins for histopathological diagnosis of Johne's disease. The new immunohistological procedures herewith detailed proved to be able to identify M. avium subsp. paratuberculosis antigens in the intestinal tissues and lymph nodes of cattle affected by either the paucibacillary or pluribacillary form of the disease. They yielded negative responses not only with healthy animals but also with those affected by tuberculosis (Mycobacterium bovis). Both immunohistological procedures proved to be as sensitive as or more sensitive than Ziehl-Neelsen staining and, in addition, to be endowed with species specificity. PMID:9665946

  4. Novel Probiotic Bifidobacterium longum subsp. infantis CECT 7210 Strain Active against Rotavirus Infections?

    PubMed Central

    Moreno Muñoz, José Antonio; Chenoll, Empar; Casinos, Beatriz; Bataller, Esther; Ramón, Daniel; Genovés, Salvador; Montava, Rebeca; Ribes, Juan Manuel; Buesa, Javier; Fàbrega, Joan; Rivero, Montserrat

    2011-01-01

    Rotavirus is the leading cause of severe acute gastroenteritis among children worldwide. It is well known that breast-feeding and vaccination afford infants protection. Since breast-feeding has drastically decreased in developed countries, efforts have been focused on the potential use of probiotics as preventive agents. In this study, a novel Bifidobacterium longum subsp. infantis strain was isolated from infant feces and selected, based on its capacity to inhibit in vitro rotavirus Wa replication (up to 36.05% infectious foci reduction) and also to protect cells from virus infection (up to 48.50% infectious foci reduction) in both MA-104 and HT-29 cell lines. Furthermore, studies using a BALB/c mouse model have proved that this strain provides preliminary in vivo protection against rotavirus infection. The strain has been deposited in the Spanish Type Culture Collection under the accession number CECT 7210. This novel strain has the main properties required of a probiotic, such as resistance to gastrointestinal juices, biliary salts, NaCl, and low pH, as well as adhesion to intestinal mucus and sensitivity to antibiotics. The food safety status has been confirmed by the absence of undesirable metabolite production and in acute ingestion studies of mice. Overall, these results demonstrate that Bifidobacterium longum subsp. infantis CECT 7210 can be considered a probiotic able to inhibit rotavirus infection. PMID:22003027

  5. The cry toxin operon of Clostridium bifermentans subsp. malaysia is highly toxic to Aedes Larval Mosquitoes.

    PubMed

    Qureshi, Nadia; Chawla, Swati; Likitvivatanavong, Supaporn; Lee, Han Lim; Gill, Sarjeet S

    2014-09-01

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

    PubMed

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

    2015-01-01

    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. PMID:26483011

  7. Photobacterium damselae subsp. damselae, a bacterium pathogenic for marine animals and humans

    PubMed Central

    Rivas, Amable J.; Lemos, Manuel L.; Osorio, Carlos R.

    2013-01-01

    Photobacterium damselae subsp. damselae (formerly Vibrio damsela) is a pathogen of a variety of marine animals including fish, crustaceans, molluscs, and cetaceans. In humans, it can cause opportunistic infections that may evolve into necrotizing fasciitis with fatal outcome. Although the genetic basis of virulence in this bacterium is not completely elucidated, recent findings demonstrate that the phospholipase-D Dly (damselysin) and the pore-forming toxins HlyApl and HlyAch play a main role in virulence for homeotherms and poikilotherms. The acquisition of the virulence plasmid pPHDD1 that encodes Dly and HlyApl has likely constituted a main driving force in the evolution of a highly hemolytic lineage within the subspecies. Interestingly, strains that naturally lack pPHDD1 show a strong pathogenic potential for a variety of fish species, indicating the existence of yet uncharacterized virulence factors. Future and deep analysis of the complete genome sequence of Photobacterium damselae subsp. damselae will surely provide a clearer picture of the virulence factors employed by this bacterium to cause disease in such a varied range of hosts. PMID:24093021

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

    PubMed Central

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

    2015-01-01

    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. PMID:26483011

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

    PubMed Central

    Sapierzy?ski, Rafa?; Szeleszczuk, Piotr

    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. PMID:24738057

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

    PubMed

    Ledwo?, Aleksandra; Sapierzy?ski, Rafa?; Augustynowicz-Kope?, 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. PMID:24738057

  11. Klebsiella pneumoniae subsp. pneumoniae–bacteriophage combination from the caecal effluent of a healthy woman

    PubMed Central

    Neve, Horst; Heller, Knut J.; Turton, Jane F.; Mahony, Jennifer; Sanderson, Jeremy D.; Hudspith, Barry; Gibson, Glenn R.; McCartney, Anne L.

    2015-01-01

    A sample of caecal effluent was obtained from a female patient who had undergone a routine colonoscopic examination. Bacteria were isolated anaerobically from the sample, and screened against the remaining filtered caecal effluent in an attempt to isolate bacteriophages (phages). A lytic phage, named KLPN1, was isolated on a strain identified as Klebsiella pneumoniae subsp. pneumoniae (capsular type K2, rmpA+). This Siphoviridae phage presents a rosette-like tail tip and exhibits depolymerase activity, as demonstrated by the formation of plaque-surrounding haloes that increased in size over the course of incubation. When screened against a panel of clinical isolates of K. pneumoniae subsp. pneumoniae, phage KLPN1 was shown to infect and lyse capsular type K2 strains, though it did not exhibit depolymerase activity on such hosts. The genome of KLPN1 was determined to be 49,037 bp (50.53 %GC) in length, encompassing 73 predicted ORFs, of which 23 represented genes associated with structure, host recognition, packaging, DNA replication and cell lysis. On the basis of sequence analyses, phages KLPN1 (GenBank: KR262148) and 1513 (a member of the family Siphoviridae, GenBank: KP658157) were found to be two new members of the genus “Kp36likevirus.” PMID:26246963

  12. The generalist Inga subnuda subsp. luschnathiana (Fabaceae): negative effect of floral visitors on reproductive success?

    PubMed

    Avila, R; Pinheiro, M; Sazima, M

    2015-05-01

    Inga species are characterised by generalist or mixed pollination system. However, this feature does not enhance reproductive rates in species with very low fruit set under natural conditions. Some ecological and genetic factors are associated with this feature, and to test the effect of massive visits on pollination success in Inga subnuda subsp. luschnathiana, we studied the efficacy of polyads deposited on stigmas of flowers isolated from visitors and polyads exposed to visitors. The proportion of polyads fixed in stigmas decreased after exposure to visitors (24 h) in comparison to stigmas isolated from visitors (hummingbirds, bees, wasps, hawkmoths and bats), and fruit set was very low. Furthermore, nectar production, sugar composition and other floral biology traits were evaluated. Increased nectar production, sugar availability and sucrose dominance during the night indicates adaptation to nocturnal visitors and supports their role as main pollinators; although the brush-flower morphology, time of anthesis, nectar dynamics and chemical composition also allow daytime visitors. Thus the species is an important resource for a diverse group of floral visitors. We conclude that excess visits (diurnal and nocturnal) are responsible for the decrease in fixed polyads in stigmas of I. subnuda subsp. luschnathiana flowers, thus contributing, with others factors, to its low fruit set. Therefore, the generalist pollination system does not result in reproductive advantages because the low fruit set in natural conditions could be the result of a negative effect of visitors/pollinators. PMID:25488371

  13. Cross-amplification of Vicia sativa subsp. sativa microsatellites across 22 other Vicia species.

    PubMed

    Raveendar, Sebastin; Lee, Gi-An; Jeon, Young-Ah; Lee, Yun Jeong; Lee, Jung-Ro; Cho, Gyu-Taek; Cho, Joon-Hyeong; Park, Jong-Hyun; Ma, Kyung-Ho; Chung, Jong-Wook

    2015-01-01

    The temperate and herbaceous genus Vicia L. is a member of the legume tribe Fabeae of the subfamily Papilionoideae. The genus Vicia comprises 166 annual or perennial species distributed mainly in Europe, Asia, and North America, but also extending to the temperate regions of South America and tropical Africa. The use of simple sequence repeat (SSR) markers for Vicia species has not been investigated as extensively as for other crop species. In this study, we assessed the potential for cross-species amplification of cDNA microsatellite markers developed from common vetch (Vicia sativa subsp. sativa). For cross-species amplification of the SSRs, amplification was carried out with genomic DNA isolated from two to eight accessions of 22 different Vicia species. For individual species or subspecies, the transferability rates ranged from 33% for V. ervilia to 82% for V. sativa subsp. nigra with an average rate of 52.0%. Because the rate of successful SSR marker amplification generally correlates with genetic distance, these SSR markers are potentially useful for analyzing genetic relationships between or within Vicia species. PMID:25608853

  14. Klebsiella pneumoniae subsp. pneumoniae-bacteriophage combination from the caecal effluent of a healthy woman.

    PubMed

    Hoyles, Lesley; Murphy, James; Neve, Horst; Heller, Knut J; Turton, Jane F; Mahony, Jennifer; Sanderson, Jeremy D; Hudspith, Barry; Gibson, Glenn R; McCartney, Anne L; van Sinderen, Douwe

    2015-01-01

    A sample of caecal effluent was obtained from a female patient who had undergone a routine colonoscopic examination. Bacteria were isolated anaerobically from the sample, and screened against the remaining filtered caecal effluent in an attempt to isolate bacteriophages (phages). A lytic phage, named KLPN1, was isolated on a strain identified as Klebsiella pneumoniae subsp. pneumoniae (capsular type K2, rmpA (+)). This Siphoviridae phage presents a rosette-like tail tip and exhibits depolymerase activity, as demonstrated by the formation of plaque-surrounding haloes that increased in size over the course of incubation. When screened against a panel of clinical isolates of K. pneumoniae subsp. pneumoniae, phage KLPN1 was shown to infect and lyse capsular type K2 strains, though it did not exhibit depolymerase activity on such hosts. The genome of KLPN1 was determined to be 49,037 bp (50.53 %GC) in length, encompassing 73 predicted ORFs, of which 23 represented genes associated with structure, host recognition, packaging, DNA replication and cell lysis. On the basis of sequence analyses, phages KLPN1 (GenBank: KR262148) and 1513 (a member of the family Siphoviridae, GenBank: KP658157) were found to be two new members of the genus "Kp36likevirus." PMID:26246963

  15. Chemical Decontamination with N-Acetyl-l-Cysteine–Sodium Hydroxide Improves Recovery of Viable Mycobacterium avium subsp. paratuberculosis Organisms from Cultured Milk

    PubMed Central

    Bradner, L.; Robbe-Austerman, S.; Beitz, D. C.

    2013-01-01

    Mycobacterium avium subsp. paratuberculosis is shed into the milk and feces of cows with advanced Johne's disease, allowing the transmission of M. avium subsp. paratuberculosis between animals. The objective of this study was to formulate an optimized protocol for the isolation of M. avium subsp. paratuberculosis in milk. The parameters investigated included chemical decontamination with N-acetyl-l-cysteine–sodium hydroxide (NALC-NaOH), alone and in combination with antibiotics (vancomycin, amphotericin B, and nalidixic acid), and the efficacy of solid (Herrold's egg yolk medium [HEY]) and liquid (Bactec 12B and para-JEM) culture media. For each experiment, raw milk samples from a known noninfected cow were inoculated with 102 to 108 CFU/ml of live M. avium subsp. paratuberculosis organisms. The results indicate that an increased length of exposure to NALC-NaOH from 5 to 30 min and an increased concentration of NaOH from 0.5 to 2.0% did not affect the viability of M. avium subsp. paratuberculosis. Additional treatment of milk samples with the antibiotics following NALC-NaOH treatment decreased the recovery of viable M. avium subsp. paratuberculosis cells more than treatment with NALC-NaOH alone. The Bactec 12B medium was the superior medium of the three evaluated for the isolation of M. avium subsp. paratuberculosis from milk, as it achieved the lowest threshold of detection. The optimal conditions for NALC-NaOH decontamination were determined to be exposure to 1.50% NaOH for 15 min followed by culture in Bactec 12B medium. This study demonstrates that chemical decontamination with NALC-NaOH resulted in a greater recovery of viable M. avium subsp. paratuberculosis cells from milk than from samples treated with hexadecylpyridinium chloride (HPC). Therefore, it is important to optimize milk decontamination protocols to ensure that low concentrations of M. avium subsp. paratuberculosis can be detected. PMID:23637290

  16. Immune Responses in Mice to Mycobacterium avium subsp. paratuberculosis Following Vaccination with a Novel 74F Recombinant Polyprotein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne’s Disease (JD) is a chronic infectious disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Here, we report the cloning and expression of a 74kDa recombinant polyprotein (Map74F) and its protective efficacy against MAP infection in mice. Map74F was generated by th...

  17. Duplex TaqMan real-time PCR assay for quantitative detection of Pantoea stewartii subsp. stewartii and Stenocarpella maydis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new TaqMan real-time PCR assay was developed for the simultaneous quantitative detection of two seedborne maize pathogens in a single assay. Pantoea stewartii subsp. stewartii (Pnss) (syn. Erwinia stewartii) is the causal agent of Stewart's bacterial wilt and leaf blight of maize. Stewart's wilt i...

  18. Draft Genome Sequence of Streptomyces ahygroscopicus subsp. wuyiensis CK-15, Isolated from Soil in Fujian Province, China

    PubMed Central

    Ge, Beibei; Liu, Yan; Liu, Binghua

    2015-01-01

    We report the first high-quality draft genome sequence of an antibiotic (wuyiencin)-producing strain, Streptomyces ahygroscopicus subsp. wuyiensis CK-15, isolated from soil samples collected from Fujian Province, China. The 9.41-Mb genome comprises 8,311 protein-coding sequences, encodes 89 structural RNAs, and shows a G+C content of 72.25%. PMID:26450721

  19. Genome Sequence of Streptococcus phocae subsp. phocae Strain ATCC 51973T Isolated from a Harbor Seal (Phoca vitulina)

    PubMed Central

    Poblete-Morales, Matías

    2015-01-01

    Streptococcus phocae subsp. phocae is a pathogen that affects different pinniped and mammalian species. This announcement reports the genome sequence of the type strain ATCC 51973 isolated in Norway from clinical specimens of harbor seal (Phoca vitulina), revealing interesting genes related to possible virulence factors. PMID:26586875

  20. Immunization with a DNA Vaccine Cocktail Induces a Th1 Response and Protects Mice Against Mycobacterium avium subsp. paratuberculosis Challenge

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  1. Complete Genome Sequence of Bacillus amyloliquefaciens subsp. plantarum CC178, a Phyllosphere Bacterium Antagonistic to Plant Pathogenic Fungi

    PubMed Central

    Kim, Byung-Yong; Lee, Sang-Yeob; Ahn, Jae-Hyung; Song, Jaekyeong; Kim, Wan-Gyu

    2015-01-01

    Bacillus amyloliquefaciens subsp. plantarum strain CC178 is a phyllosphere bacterium with antagonistic activity against a wide range of plant fungal pathogens. The genome of strain CC178 is 3,916,828 bp in size and harbors 3,972 genes. Six giant gene clusters are dedicated to the nonribosomal synthesis of antimicrobial polypeptides and polyketides. PMID:25573933

  2. Selection of single chain variable fragments (scFv) against Xylella fastidiosa subsp. pauca by phage display

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Xylella fastidiosa is a gram-negative member of the gamma proteobacteria. Xylella fastidiosa subsp pauca causes citrus variegated chlorosis in Brazil and enjoys ‘select agent’ status in the United States. Antibody based detection assays are commercially available for Xylella fastidiosa, and are ef...

  3. Progress in the Development of Crimson Sweet-type Watermelon Breeding Lines with Resistance to Acidovorax Avenae Subsp. Citrulli

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial fruit blotch (Acidovorax avenae subsp. citrulli [Schaad et al.] Willems et al.) continues to occur almost every year and has the potential to cause a disaster for the watermelon industry. In this study, Crimson Sweet watermelon was crossed with PI482279 and PI494817, two Citrullus lanatus...

  4. Identification and Functional Characterization of the Iron-dependent Regulator (IdeR) of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP), a ruminant pathogen, has unique iron requirements based on the observation that it is mycobactin dependent for successful cultivation in vitro. Thus an elucidation of iron regulation in MAP is expected to provide an understanding of its survival in ...

  5. Complete Genome Sequence of Bacillus amyloliquefaciens subsp. plantarum CC178, a Phyllosphere Bacterium Antagonistic to Plant Pathogenic Fungi.

    PubMed

    Kim, Byung-Yong; Lee, Sang-Yeob; Ahn, Jae-Hyung; Song, Jaekyeong; Kim, Wan-Gyu; Weon, Hang-Yeon

    2015-01-01

    Bacillus amyloliquefaciens subsp. plantarum strain CC178 is a phyllosphere bacterium with antagonistic activity against a wide range of plant fungal pathogens. The genome of strain CC178 is 3,916,828 bp in size and harbors 3,972 genes. Six giant gene clusters are dedicated to the nonribosomal synthesis of antimicrobial polypeptides and polyketides. PMID:25573933

  6. QTL mapping for salt tolerance and domestication-related traits in Vigna marina subsp. oblonga, a halophytic species.

    PubMed

    Chankaew, Sompong; Isemura, Takehisa; Naito, Ken; Ogiso-Tanaka, Eri; Tomooka, Norihiko; Somta, Prakit; Kaga, Akito; Vaughan, Duncan A; Srinives, Peerasak

    2014-03-01

    QTL mapping in F2 population [V. luteola × V. marina subsp. oblonga] revealed that the salt tolerance in V. marina subsp. oblonga is controlled by a single major QTL. The habitats of beach cowpea (Vigna marina) are sandy beaches in tropical and subtropical regions. As a species that grows closest to the sea, it has potential to be a gene source for breeding salt-tolerant crops. We reported here for the first time, quantitative trait loci (QTLs) mapping for salt tolerance in V. marina. A genetic linkage map was constructed from an F2 population of 120 plants derived from an interspecific cross between V. luteola and V. marina subsp. oblonga. The map comprised 150 SSR markers. The markers were clustered into 11 linkage groups spanning 777.6 cM in length with a mean distance between the adjacent markers of 5.59 cM. The F2:3 population was evaluated for salt tolerance under hydroponic conditions at the seedling and developmental stages. Segregation analysis indicated that salt tolerance in V. marina is controlled by a few genes. Multiple interval mapping consistently identified one major QTL which can explain about 50% of phenotypic variance. The flanking markers may facilitate transfer of the salt tolerance allele from V. marina subsp. oblonga into related Vigna crops. The QTL for domestication-related traits from V. marina are also discussed. PMID:24370961

  7. Evolutionary trends in two strains of Salmonella enterica subsp. I serovar Enteritidis PT13a that vary in virulence potential

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. I serovar Enteritidis (S. Enteritidis) is the world's leading cause of salmonellosis. Eggs contaminated by apparently healthy hens and that have been improperly cooked can result in illness in humans who consume them. Although the incidence of this pathogen within the Uni...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  9. Survival of the Causative Agent of Johne’s Disease (Mycobacterium avium subsp. paratuberculosis) in Biofilms on Trough Materials

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The continued global increase in the number of cases of Johne’s disease among dairy cattle suggests that there remain hidden sources of contamination in the farm environment where susceptible animals may be routinely exposed to Mycobacterium avium subsp. paratuberculosis (MAP), the causative agent o...

  10. Macrophage Transcriptional Response to Species-Adapted Mycobacterium avium subsp. Paratuberculosis Isolates: The Role of Pathogen Genotype in Host Response

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The transcriptional response of human and bovine macrophages to Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) isolates from cattle and sheep were examined using DNA microarrays. M. paratuberculosis is the etiologic agent of Johne’s Disease, a chronic infection of ruminant anima...

  11. Evaluation of survival of Mycobacterium avium subsp. paratuberculosis (Map) in ciliates isolated from Johne’s positive cow.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Persistence of Mycobacterium avium subsp. paratuberculosis (Map) in farm environments is not well understood. Previously we examined the ability of amoebae from a cow’s watering trough to sequester and enhance growth of Map and found that one amoeba species released vesicles containin...

  12. Complete Genome Sequences of Two Outbreak Strains of Salmonella enterica subsp. enterica Serovar Thompson Associated with Cilantro

    PubMed Central

    Huynh, Steven; Gorski, Lisa; Cooper, Kerry K.; Miller, William G.

    2015-01-01

    Salmonella enterica subsp. enterica serovar Thompson strains RM1984 (CADPH-99A2334) and RM1986 (CADPH-99A2345) are associated with a 1999 outbreak in contaminated cilantro. We report here the complete genome sequences and annotation of these two S. Thompson strains. These genomes are distinct and provide additional data for our understanding of S. enterica. PMID:26586897

  13. Comparison of fecal DNA extraction kits for the detection of Mycobacterium avium subsp. paratuberculosis by polymerase chain reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Culture of Mycobacterium avium subsp. paratuberculosis (MAP) from feces has been considered the gold standard for the diagnosis of paratuberculosis for many years. However, direct fecal PCR is becoming more widely used today, demonstrating similar sensitivity and specificity to culture. To ensure ef...

  14. Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Give, Isolated from an Imported Chili Powder Product

    PubMed Central

    Chen, Yi; Ayers, Sherry; Melka, David; Laasri, Anna; Payne, Justin S.; Zheng, Jie; Son, Insook; Timme, Ruth; Kastanis, George; Hammack, Thomas S.; Strain, Errol; Allard, Marc W.; Evans, Peter S.; Brown, Eric W.

    2015-01-01

    We report the genome sequence of Salmonella enterica subsp. enterica serovar Give (CFSAN012622), isolated from imported chili powder in 2014. This genome contains genes previously reported to be specific only to S. enterica serovar Enteritidis. This strain shows a unique pulsed-field gel electrophoresis (PFGE) pattern clustering with serovar Enteritidis (JEG X01.0005). PMID:26139723

  15. A LONG TERM, SUB-CLINICAL, OUTBREAK OF SALMONELLA ENTERICA SUBSP. ENTERICA CERRO IN A PENNSYLVANIA DAIRY HERD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella is a prominent foodborne pathogen in the US and dairy cattle have frequently been identified as reservoirs for this organism. Here we describe a long term outbreak of Salmonella enterica subsp. enterica Cerro that was detected in a Pennsylvania dairy herd during routine monitoring as pa...

  16. Evolutionary trends in two strains of Salmonella enterica subsp. I serovar Enteritidis PT13a that vary in virulence potential.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. I serovar Enteritidis (S. Enteritidis) is the world's leading cause of salmonellosis. Eggs that are contaminated by apparently healthy hens and that have been improperly cooked can result in illness in humans who consume them. Although the incidence of this pathogen within...

  17. PHYLOGENETIC STUDIES OF CORN AND RICE STRAINS OF ACIDOVORAX AVENAE SUBSP. AVENAE BY DNA/DNA HYBRIDIZATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acidovorax avenae subsp. avenae (Aaa) is the causal agent of diseases of several important economic crops, including bacterial streak of corn (Zea mays) and bacterial stripe of rice (Oryza sativa). To determine the phylogenetic relationship of these two pathogens, a highly reproducible S1 exonuclea...

  18. Parturition Invokes Changes in Peripheral blood Mononuclear Cell Populations in Holstein Dairy Cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is estimated to infect more than 22% of US dairy herds. Once infected, cows may remain in the asymptomatic subclinical state until a period of stress, such as parturition. Parturition has a major impact on the number of ...

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

    PubMed Central

    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

  20. Shedding of Mycobacterium avium subsp. paratuberculosis into milk and colostrum of naturally infected dairy cows over complete lactation cycles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The primary mode of transmission of Mycobacterium avium subsp. paratuberculosis (MAP) is fecal-oral. However, MAP is also shed into the milk and colostrum of infected cows. The objective of this study was to identify if an association exists between stage of MAP infection and days in lactation with ...

  1. Optimization of methods for the detection of Mycobacterium avium subsp. paratuberculosis in milk and colostrum of naturally infected dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two decontamination chemicals, hexadecylpyridinium choride (HPC) and N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH), were compared for their efficacy of reducing the growth of non-specific microorganisms in milk while minimally affecting the recovery of Mycobacterium avium subsp. paratuberculosis ...

  2. Optimization of methods for the detection of Mycobacterium avium subsp. paratuberculosis in milk and colostrum of naturally infected dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) is primarily shed into the feces but it has also been isolated from the milk and colostrum of cows. Because of this, there exists concern about transfer of the organism from dam to calf and about the prevalence of MAP in the milk supply. The prevalen...

  3. Genome Sequence of Lactobacillus delbrueckii subsp. lactis CNRZ327, a Dairy Bacterium with Anti-Inflammatory Properties

    PubMed Central

    El Kafsi, Hela; Binesse, Johan; Loux, Valentin; Buratti, Julien; Boudebbouze, Samira; Dervyn, Rozenn; Hammani, Amal; Maguin, Emmanuelle

    2014-01-01

    Lactobacillus delbrueckii subsp. lactis CNRZ327 is a dairy bacterium with anti-inflammatory properties both in vitro and in vivo. Here, we report the genome sequence of this bacterium, which appears to contain no less than 215 insertion sequence (IS) elements, an exceptionally high number regarding the small genome size of the strain. PMID:25035318

  4. Genome Sequence of the Lactic Acid Bacterium Lactococcus lactis subsp. lactis TOMSC161, Isolated from a Nonscalded Curd Pressed Cheese

    PubMed Central

    Velly, H.; Abraham, A.-L.; Loux, V.; Delacroix-Buchet, A.; Fonseca, F.; Bouix, M.

    2014-01-01

    Lactococcus lactis is a lactic acid bacterium used in the production of many fermented foods, such as dairy products. Here, we report the genome sequence of L. lactis subsp. lactis TOMSC161, isolated from nonscalded curd pressed cheese. This genome sequence provides information in relation to dairy environment adaptation. PMID:25377704

  5. Genome Sequence of the Lactic Acid Bacterium Lactococcus lactis subsp. lactis TOMSC161, Isolated from a Nonscalded Curd Pressed Cheese.

    PubMed

    Velly, H; Renault, P; Abraham, A-L; Loux, V; Delacroix-Buchet, A; Fonseca, F; Bouix, M

    2014-01-01

    Lactococcus lactis is a lactic acid bacterium used in the production of many fermented foods, such as dairy products. Here, we report the genome sequence of L. lactis subsp. lactis TOMSC161, isolated from nonscalded curd pressed cheese. This genome sequence provides information in relation to dairy environment adaptation. PMID:25377704

  6. Genome Sequence of Lactobacillus delbrueckii subsp. lactis CNRZ327, a Dairy Bacterium with Anti-Inflammatory Properties.

    PubMed

    El Kafsi, Hela; Binesse, Johan; Loux, Valentin; Buratti, Julien; Boudebbouze, Samira; Dervyn, Rozenn; Hammani, Amal; Maguin, Emmanuelle; van de Guchte, Maarten

    2014-01-01

    Lactobacillus delbrueckii subsp. lactis CNRZ327 is a dairy bacterium with anti-inflammatory properties both in vitro and in vivo. Here, we report the genome sequence of this bacterium, which appears to contain no less than 215 insertion sequence (IS) elements, an exceptionally high number regarding the small genome size of the strain. PMID:25035318

  7. Sequencing of the complete genome of an araphid pennate diatom Synedra acus subsp. radians from Lake Baikal.

    PubMed

    Galachyants, Y P; Zakharova, Yu R; Petrova, D P; Morozov, A A; Sidorov, I A; Marchenkov, A M; Logacheva, M D; Markelov, M L; Khabudaev, K V; Likhoshway, Ye V; Grachev, M A

    2015-01-01

    High-throughput method of sequencing was applied to determine the complete nucleotide sequence of an araphid pennate diatom Synedra acus subsp. radians from Lake Baikal (East Siberia). The assembled genome has a total length of 98 Mbp, the mean coverage is 33x. Structure-functional annotation of the genome was performed. PMID:25937221

  8. ZAP-70, CTLA-4, and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  9. Evaluation of Control Points in Youngstock and Adult Dairy Cow Management to Control Transmission of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Goal Complete a series of controlled on-farm trials to critically evaluate the efficacy and cost-benefit of commonly recommended management practices for reducing the transmission of Mycobacterium avium subsp. paratuberculosis (Map) in infected herds. Objective 1. Evaluate the effect of maternity...

  10. Genome Sequences of Strain ATCC 29281 and Pin and Northern Red Oak Isolates of Lonsdalea quercina subsp. quercina

    PubMed Central

    Ibarra Caballero, Jorge; Zerillo, Marcelo M.; Snelling, Jacob; Cranshaw, Whitney; Boucher, Christina

    2014-01-01

    Two bacteria identified as Lonsdalea quercina subsp. quercina were isolated from oak trees showing symptoms of drippy blight. Here, we present their draft genome assemblies, as well as that of the type strain of this species. To our knowledge, these are the first published genome sequences of this subspecies of Lonsdalea quercina. PMID:24926062

  11. ZAP-70, CTLA-4 and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  12. Evolutionary trends in two strains of Salmonella enterica subsp. I serovar Enteritidis PT13a that vary in virulence potential.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. I serovar Enteritidis (S. Enteritidis) is the world's leading cause of salmonellosis. Eggs contaminated by apparently healthy hens and that have been improperly cooked can result in illness in humans who consume them. Although the incidence of this pathogen within the Unit...

  13. Evolutionary trends in two strains of Salmonella enterica subsp. I serovar Enteritidis PT13a that vary in virulence potential.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica subsp. I serovar Enteritidis (S. Enteritidis) is the world's leading cause of salmonellosis. Eggs contaminated by apparently healthy hens can result in illness in humans who consume them. Although the incidence of this pathogen within the United States has not been as high as it ...

  14. Adult invasive and noninvasive infections due to Streptococcus dysgalactiae subsp. equisimilis in France from 2006 to 2010.

    PubMed

    Loubinoux, Julien; Plainvert, Céline; Collobert, Gislène; Touak, Gérald; Bouvet, Anne; Poyart, Claire

    2013-08-01

    We characterized 182 Streptococcus dysgalactiae subsp. equisimilis isolates and analyzed the epidemiological data on the corresponding infections. stG6, stG485, and stG6792 were the 3 most prevalent invasive emm types among the 27 different emm types recovered. High rates of antimicrobial resistance were observed for macrolides (26.4%) and tetracycline (34.6%). PMID:23698531

  15. Deciphering the conserved genetic loci implicated in plant disease control through comparative genomics of Bacillus amyloliquefaciens subsp. plantarum

    PubMed Central

    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

  16. Composition, Enantiomeric Distribution and Antimicrobial Activity of the Essential Oil of Tanacetum argenteum subsp. flabellifolium Essential Oil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tanacetum argenteum (Lam.) Willd. subsp. flabellifolium (Boiss. & Heldr.) Grierson of Asteraceae is an endemic species in Turkey. Hydrodistillation of aerial parts using a Clevenger apparatus yielded an essential oil, which was subsequently analyzed by gas chromatography/mass spectroscopy (GC/MS). ...

  17. Rapid Assessment of the Viability of Mycobacterium avium subsp. paratuberculosis Cells after Heat Treatment, Using an Optimized Phage Amplification Assay?

    PubMed Central

    Foddai, Antonio; Elliott, Christopher T.; Grant, Irene R.

    2010-01-01

    Thermal inactivation experiments were carried out to assess the utility of a recently optimized phage amplification assay to accurately enumerate viable Mycobacterium avium subsp. paratuberculosis cells in milk. Ultra-heat-treated (UHT) whole milk was spiked with large numbers of M. avium subsp. paratuberculosis organisms (106 to 107 CFU/ml) and dispensed in 100-?l aliquots in thin-walled 200-?l PCR tubes. A Primus 96 advanced thermal cycler (Peqlab, Erlangen, Germany) was used to achieve the following time and temperature treatments: (i) 63°C for 3, 6, and 9 min; (ii) 68°C for 20, 40, and 60 s; and (iii) 72°C for 5, 10, 15, and 25 s. After thermal stress, the number of surviving M. avium subsp. paratuberculosis cells was assessed by both phage amplification assay and culture on Herrold's egg yolk medium (HEYM). A high correlation between PFU/ml and CFU/ml counts was observed for both unheated (r2 = 0.943) and heated (r2 = 0.971) M. avium subsp. paratuberculosis cells. D and z values obtained using the two types of counts were not significantly different (P > 0.05). The D68°C, mean D63°C, and D72°C for four M. avium subsp. paratuberculosis strains were 81.8, 9.8, and 4.2 s, respectively, yielding a mean z value of 6.9°C. Complete inactivation of 106 to 107 CFU of M. avium subsp. paratuberculosis/ml milk was not observed for any of the time-temperature combinations studied; 5.2- to 6.6-log10 reductions in numbers were achieved depending on the temperature and time. Nonlinear thermal inactivation kinetics were consistently observed for this bacterium. This study confirms that the optimized phage assay can be employed in place of conventional culture on HEYM to speed up the acquisition of results (48 h instead of a minimum of 6 weeks) for inactivation experiments involving M. avium subsp. paratuberculosis-spiked samples. PMID:20097817

  18. Suspicion of Mycobacterium avium subsp. paratuberculosis Transmission between Cattle and Wild-Living Red Deer (Cervus elaphus) by Multitarget Genotyping

    PubMed Central

    Fritsch, Isabel; Luyven, Gabriele; Köhler, Heike; Lutz, Walburga

    2012-01-01

    Multitarget genotyping of the etiologic agent Mycobacterium avium subsp. paratuberculosis is necessary for epidemiological tracing of paratuberculosis (Johne's disease). The study was undertaken to assess the informative value of different typing techniques and individual genome markers by investigation of M. avium subsp. paratuberculosis transmission between wild-living red deer and farmed cattle with known shared habitats. Fifty-three M. avium subsp. paratuberculosis type II isolates were differentiated by short sequence repeat analysis (SSR; 4 loci), mycobacterial interspersed repetitive-unit–variable-number tandem-repeat analysis (MIRU-VNTR; 8 loci), and restriction fragment length polymorphism analysis based on IS900 (IS900-RFLP) using BstEII and PstI digestion. Isolates originated from free-living red deer (Cervus elaphus) from Eifel National Park (n = 13), six cattle herds living in the area of this park (n = 23), and five cattle herds without any contact with these red deer (n = 17). Data based on individual herds and genotypes verified that SSR G2 repeats did not exhibit sufficient stability for epidemiological studies. Two common SSR profiles (without G2 repeats), nine MIRU-VNTR patterns, and nine IS900-RFLP patterns were detected, resulting in 17 genotypes when combined. A high genetic variability was found for red deer and cattle isolates within and outside Eifel National Park, but it was revealed only by combination of different typing techniques. Results imply that within this restricted area, wild-living and farmed animals maintain a reservoir for specific M. avium subsp. paratuberculosis genotypes. No host relation of genotypes was obtained. Results suggested that four genotypes had been transmitted between and within species and that one genotype had been transmitted between cattle herds only. Use of multitarget genotyping for M. avium subsp. paratuberculosis type II strains and sufficiently stable genetic markers is essential for reliable interpretations of epidemiological studies on paratuberculosis. PMID:22179249

  19. Utility of Sequencing the erm(41) Gene in Isolates of Mycobacterium abscessus subsp. abscessus with Low and Intermediate Clarithromycin MICs

    PubMed Central

    Vasireddy, Sruthi; Vasireddy, Ravikiran; Iakhiaeva, Elena; Howard, Susan T.; Nash, Kevin; Parodi, Nicholas; Strong, Anita; Gee, Martha; Smith, Terry; Wallace, Richard J.

    2015-01-01

    The erm(41) gene confers inducible macrolide resistance in Mycobacterium abscessus subsp. abscessus, calling into question the usefulness of macrolides for treating M. abscessus subsp. abscessus infections. With an extended incubation (14 days), isolates with MICs of ?8 ?g/ml are considered macrolide resistant by current CLSI guidelines. Our goals were to determine the incidence of macrolide susceptibility in U.S. isolates, the validity of currently accepted MIC breakpoints, and the erm(41) sequences associated with susceptibility. Of 349 isolates (excluding those with 23S rRNA gene mutations), 85 (24%) had clarithromycin MICs of ?8 ?g/ml. Sequencing of the erm(41) genes from these isolates, as well as from isolates with MICs of ?16 ?g/ml, including ATCC 19977T, revealed 10 sequevars. The sequence in ATCC 19977T was designated sequevar (type) 1; most macrolide-resistant isolates were of this type. Seven sequevars contained isolates with MICs of >16 ?g/ml. The T28C substitution in erm(41), previously associated with macrolide susceptibility, was identified in 62 isolates (18%) comprising three sequevars, with MICs of ?2 (80%), 4 (10%), and 8 (10%) ?g/ml. No other nucleotide substitution was associated with macrolide susceptibility. We recommend that clarithromycin susceptibility breakpoints for M. abscessus subsp. abscessus be changed from ?2 to ?4 ?g/ml and that isolates with an MIC of 8 ?g/ml have repeat MIC testing or erm sequencing performed. Our studies suggest that macrolides are useful for treating approximately 20% of U.S. isolates of M. abscessus subsp. abscessus. Sequencing of the erm gene of M. abscessus subsp. abscessus will predict inducible macrolide susceptibility. PMID:25653399

  20. Modulation of Cytokine Expression and Lymphocyte Subsets During the Periparturient Period in Dairy Cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    On-farm observations suggest that dairy cows infected with Mycobacterium avium subsp. paratuberculosis (MAP) may demonstrate increased signs of clinical disease during the weeks following parturition. To date, limited research is available characterizing host immunity in periparturient dairy cows ...

  1. Treatment with antibiotics is detrimental to the recovery of viable Mycobacterium avium subsp. paratuberculosis cultured from milk and colostrum of dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Antibiotic cocktails are frequently used as secondary decontaminants prior to the culture of Mycobacterium avium subsp. paratuberculosis (MAP). This study investigated whether secondary incubation with an antibiotic cocktail containing vancomycin, nalidixic acid, and amphotericin B after primary exp...

  2. Complete Genome Sequence of Salmonella enterica subsp. enterica Serovar Typhi Isolate B/SF/13/03/195 Associated with a Typhoid Carrier in Pasir Mas, Kelantan, Malaysia.

    PubMed

    Muhamad Harish, Salwani; Sim, Kee-Shin; Mohd Nor, Fauziah; Mat Hussin, Hani; Hamzah, Wan Mansor; Najimudin, Nazalan; Aziah, Ismail

    2015-01-01

    We report here the complete genome sequence of Salmonella enterica subsp. enterica serovar Typhi B/SF/13/03/195 obtained from a typhoid carrier, who is a food handler in Pasir Mas, Kelantan. PMID:26564035

  3. Effect of Feeding Heat-Treated Colostrum on Preweaning Health, Economics and Transmission of Mycobacterium avium subsp. paratuberculosis in Dairy Calves: Phase I

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction and Objectives Colostrum provides protective immunoglobulins (Ig) and nutrients essential for calf health and performance. However, colostrum may also represent an early source of pathogen exposure including Mycobacterium avium subsp. paratuberculosis (MAP). Pilot studies have suggest...

  4. Granuloma Coinfection with Mycobacterium bovis, Mycobacterium avium subsp. paratuberculosis, and Corynebacterium pseudotuberculosis in Five Hunted Red deer (Cervus elaphus) in Portugal.

    PubMed

    Matos, Ana C; Dias, Ana P; Morais, Márcia; Figueira, Luís; Martins, Maria H; Matos, Manuela; Pinto, Maria L; Coelho, Ana C

    2015-07-01

    We report granulomatous lymphadenitis in red deer (Cervus elaphus) in Portugal caused by coinfection with Corynebacterium pseudotuberculosis, Mycobacterium bovis, and Mycobacterium avium subsp. paratuberculosis, as demonstrated by molecular methods. PMID:25932668

  5. Production and Evaluation of an Improved Mycobacterium avium subsp. paratuberculosis Purified Protein Derivative for Use in In-Vivo and In-Vitro Diagnostic Testing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Purified protein derivatives (PPD’s) were prepared from the cultured filtrate of Mycobacterium avium subsp. paratuberculosis (MAP) ATCC strain 19698. Production of PPD has historically been problematic for maintaining optimal floating cultures yielding defined immunogenic components. To obtain mor...

  6. Genome Sequence of Salmonella enterica subsp. enterica Serovar Typhi Isolate PM016/13 from Untreated Well Water Associated with a Typhoid Outbreak in Pasir Mas, Kelantan, Malaysia.

    PubMed

    Muhamad Harish, Salwani; Sim, Kee-Shin; Najimudin, Nazalan; Aziah, Ismail

    2015-01-01

    Salmonella enterica subsp. enterica serovar Typhi is a human-restricted pathogen that causes typhoid fever. Even though it is a human-restricted pathogen, the bacterium is also isolated from environments such as groundwater and pond water. Here, we describe the genome sequence of the Salmonella enterica subsp. enterica serovar Typhi PM016/13 which was isolated from well water during a typhoid outbreak in Kelantan, Malaysia, in 2013. PMID:26564032

  7. Aeromonas salmonicida subsp. salmonicida in the light of its type-three secretion system

    PubMed Central

    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

  8. Antioxidant and antiproliferative activity of Hypericum hircinum L. subsp. majus (Aiton) N. Robson essential oil.

    PubMed

    Quassinti, Luana; Lupidi, Giulio; Maggi, Filippo; Sagratini, Gianni; Papa, Fabrizio; Vittori, Sauro; Bianco, Armandodoriano; Bramucci, Massimo

    2013-01-01

    This study was undertaken to assess the antioxidant and antiproliferative potential of the essential oil of Hypericum hircinum L. subsp. majus (Aiton) N. Robson. Analysis of the oil composition revealed that sesquiterpene hydrocarbons (69.3%) dominate, cis-?-guaiene, ?-selinene and (E)-caryophyllene being the most representative. Significant values of antioxidant activity were found using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assays. The essential oil revealed antiproliferative activity as evaluated on human glioblastoma (T98G), human prostatic adenocarcinoma (PC3), human squamous carcinoma (A431) and mouse melanoma (B16-F1) tumour cell lines by MTT assay. PMID:22480321

  9. Development of an immunochromatographic strip for rapid detection of Pantoea stewartii subsp. stewartii.

    PubMed

    Feng, Min; Kong, Dezhao; Wang, Wenbing; Liu, Liqiang; Song, Shanshan; Xu, Chuanlai

    2015-01-01

    A rapid, simple, sensitive, and specific immunochromatographic test strip was developed for the detection of Pantoea stewartii subsp. stewartii (Pss) in corn seed which was soaked overnight and then centrifuged for precipitate re-dissolved as samples. A pair of sensitive monoclonal antibodies for the immunochromatographic test strip was generated by mice immunization and cell fusion. Under optimized conditions, the lower detection limit of the strips for Pss was 1 × 10(5) cfu/mL both in 0.01 M phosphate buffer solution and corn seed samples, with no cross-reactivity with other common plant pathogens. The developed strip is useful and rapid for the detection of Pss in corn seed samples. PMID:25686315

  10. [Possible association between Mycobacterium avium subsp paratuberculosis infection and Crohn's disease].

    PubMed

    Retamal, Patricio; Beltrán, Caroll; Abalos, Pedro; Quera, Rodrigo; Hermoso, Marcela

    2011-06-01

    Paratuberculosis is a chronic intestinal disease of animals caused by Mycobacterium avium subsp. paratuberculosis (MAP), which has some pathological features similar to Crohn's disease (CD) in humans. The presence of MAP in food for human consumption and in affected tissues of patients with CD has been detected. Therefore, a causal association between this microorganism and the disease in humans, has been postulated. However, several related studies have failed to confirm this hypothesis and the scientific acceptance of MAP as a zoonotic agent remains controversial. This review presents the main findings related to this issue, contrasting evidences for and against an association between MAP and CD. The need to promote national studies focusing on this area is suggested. PMID:22051762

  11. Chemomicin A, a new angucyclinone antibiotic produced by Nocardia mediterranei subsp. kanglensis 1747-64.

    PubMed

    Sun, Cheng-Hang; Wang, Yue; Wang, Zheng; Zhou, Jian-Qin; Jin, Wen-Zao; You, Xue-Fu; Gao, Hong; Zhao, Li-Xun; Si, Shu-Yi; Li, Xue

    2007-03-01

    A new angucyclinone antibiotic, chemomicin A was isolated from cultured broth of Nocardia mediterranei subsp. kanglensis 1747-64. Its chemical structure was determined to be 1,2,3,4a,5,6,6a,12a,12b-nonahydro-1,2,3,8,12,12b-hexahydroxy-3-methyl-6a,12a-epoxybenz[a]anthracen-4,7(12H)-dione by a detailed spectroscopic analysis. Chemomicin A had antimicrobial activity against Bacillus subtilis and Enterococcus faecium with MIC values of 10.2 and 20.4 microM, respectively, and showed cytotoxicity against human colorectal cancer HCT116 cells and human esophageal carcinoma YES-2 cells with IC50 values of 127 and 153 microM, respectively. PMID:17446695

  12. Infection of the thyroid gland caused by Campylobacter fetus subsp. fetus.

    PubMed

    Goegebuer, T; Verhaeghe, J P; Verlinde, A; De Laere, E; Surmont, I

    2007-01-01

    We report what we consider to be the first case of an abscess of the thyroid gland due to Campylobacter fetus subsp. fetus (C. fetus) in a patient suffering from hyperthyroidism. C. fetus is known as a rare and opportunistic pathogen in humans, causing a broad variety of systemic infections. Acquisition by humans is thought to occur through contact with animals or animal products and to start as a gastro-intestinal colonization. The detection of C. fetus in stool is challenging, since culture efforts are generally directed in order to fulfil growth requirements of C.jejuni, a much more common enteric pathogen. Detection of C. fetus in non-stool samples is even more challenging since routine culture doesn't imply prolonged incubation (>72h), selective media and microaerophilic conditions. It is therefore not unlikely that human infections caused by C. fetus occur more often than generally assumed. PMID:17547296

  13. Haemorrhagic septicaemia by Aeromonas salmonicida subsp. salmonicida in a black-tip reef shark (Carcharhinus melanopterus).

    PubMed

    Briones, V; Fernández, A; Blanco, M; Ramiro, F; de Vicente, M L; García, J; Méndez, J L; Goyache, J

    1998-09-01

    One black-tip reef shark (Carcharhinus melanopterus) was found dead without previous signs of disease. Major lesions consisted in cutaneous erythema, mainly at the base of the fins, focal to diffuse inflammatory lesions in gills and intestinal wall, and discrete haemorrhages in the same organs, liver and kidneys. Microcolonies of Gram-negative rods were observed in the lamina propia of gills, underneath the intestinal mucosa and randomly distributed in the renal and hepatic parenchymas. Also, emboli containing Gram-negative rods were observed in capillaries of these organs. Aeromonas salmonicida subsp. salmonicida was isolated in pure culture from gills, liver and intestine. Specific immunostaining confirmed the relationship between the isolate and lesion-associated bacteria. No previous reports on this infection in sharks have been found in the literature. PMID:9780832

  14. A new sesquiterpene lactone and other constituents of Moquiniastrum polymorphum subsp. floccosum (Asteraceae).

    PubMed

    Strapasson, L B; Rúdiger, André Luis; Burrow, Robert A; Barison, Andersson; Stefanello, Maria Elida A

    2014-11-01

    A new guaianolide, 1S, 3S, 5R, 6S, 7S, 11R-l-hydroxy-11?,13-dihydrozaluzanin C (1), was isolated from Moquiniastrum polymorphum subsp. floccosum trunk bark, together with fifteen known compounds, which were identified as 11?,13-dihydroglucozaluzanin C (2), 8?-hydroxy-11?,13-dihydrozaluzanin C (3), zaluzanin C (4), gochnatiolide B (5), ethyl caffeate (6), methyl chlorogenate (7), ethyl chlorogenate (8), methyl 3,5-dicaffeoyl quinate (9), ethyl 3,5-dicaffeoyl quinate (10), methyl 4,5-dicaffeoyl quinate (11), ethyl 4,5-dicaffeoyl quinate (12), ethyl 3,4-dicaffeoyl quinate (13), 3,5-dicaffeoyl quinic acid (14), 4,5-dicaffeoyl quinic acid (15), and 3,4-dicaffeoyl quinic acid (16). With the exception of 5, all known compounds are being reported for the first time in M. polymorphum. PMID:25532275

  15. Development of Loop-Mediated Isothermal Amplification for Detection of Leifsonia xyli subsp. xyli in Sugarcane

    PubMed Central

    Liu, Jing; Guo, Jinlong; Chen, Rukai; Grisham, Michael Paul

    2013-01-01

    Ratoon stunt, caused by the xylem-limited coryneform bacterium Leifsonia xyli subsp. xyli (Lxx), is a deep bacteriosis and prevalent in most of sugarcane-producing countries. Based on loop-mediated isothermal amplification (LAMP), we developed a method for detecting Lxx. The major advantages of the LAMP method are visual judgment by color and time saving with only 60?min for identification of Lxx and without the need for costly PCR apparatus and gel scanner. In the present study, positive and negative samples detected by the LAMP method were clearly distinguishable. When total DNA extracted from internode juice was used as the template, the sensitivity of LAMP was 10 times higher than that of the conventional PCR detection. The LAMP assay is a highly specific, rapid, and sensitive method for the diagnosis of ratoon stunt caused by Lxx in sugarcane. This is the first report of LAMP-based assay for the detection of Lxx in sugarcane. PMID:23710444

  16. Association between cattle herd Mycobacterium avium subsp. paratuberculosis (MAP) infection and infection of a hare population.

    PubMed

    Salgado, Miguel; Monti, Gustavo; Sevilla, Iker; Manning, Elizabeth

    2014-10-01

    Paratuberculosis has long been considered a disease of domestic and wild ruminants only. The known host range of Mycobacterium avium subsp. paratuberculosis (MAP) was recently extended to include non-ruminant wildlife species believed to be exposed to spillover of MAP from infected domestic cattle herds. The aim of the present study was to assess the association between cattle herd MAP infection pressure level and the infection level of a hare population in two dairy farms of southern Chile. Fifty hares from a herd A and 42 hares from herd B were captured and sampled for MAP culture. The results showed a statistically significant association between the cattle herds' infection prevalence and the hare infection prevalence. PMID:25030465

  17. Draft genome sequence of Francisella tularensis subsp. holarctica BD11-00177.

    PubMed

    Coolen, Jordy P M; Sjödin, Andreas; Maraha, Boulos; Hajer, Gerard F; Forsman, Mats; Verspui, Ellen; Frenay, Hendrina M E; Notermans, Daan W; de Vries, Maaike C; Reubsaet, Frans A G; Paauw, Armand; Roeselers, Guus

    2013-07-30

    Francisella tularensis is a facultative intracellular bacterium in the class Gammaproteobacteria. This strain is of interest because it is the etiologic agent of tularemia and a highly virulent category A biothreat agent. Here we describe the draft genome sequence and annotation of Francisella tularensis subsp. holarctica BD11-00177, isolated from the first case of indigenous tularemia detected in The Netherlands since 1953. Whole genome DNA sequence analysis assigned this isolate to the genomic group B.FTNF002-00, which previously has been exclusively reported from Spain, France, Italy, Switzerland and Germany. Automatic annotation of the 1,813,372 bp draft genome revealed 2,103 protein-coding and 46 RNA genes. PMID:24501637

  18. Phylogenomics of Brazilian epidemic isolates of Mycobacterium abscessus subsp. bolletii reveals relationships of global outbreak strains.

    PubMed

    Davidson, Rebecca M; Hasan, Nabeeh A; de Moura, Vinicius Calado Nogueira; Duarte, Rafael Silva; Jackson, Mary; Strong, Michael

    2013-12-01

    Rapidly growing, non-tuberculous mycobacteria (NTM) in the Mycobacterium abscessus (MAB) species are emerging pathogens that cause various diseases including skin and respiratory infections. The species has undergone recent taxonomic nomenclature refinement, and is currently recognized as two subspecies, M. abscessus subsp. abscessus (MAB-A) and M. abscessus subsp. bolletii (MAB-B). The recently reported outbreaks of MAB-B in surgical patients in Brazil from 2004 to 2009 and in cystic fibrosis patients in the United Kingdom (UK) in 2006 to 2012 underscore the need to investigate the genetic diversity of clinical MAB strains. To this end, we sequenced the genomes of two Brazilian MAB-B epidemic isolates (CRM-0019 and CRM-0020) derived from an outbreak of skin infections in Rio de Janeiro, two unrelated MAB strains from patients with pulmonary infections in the United States (US) (NJH8 and NJH11) and one type MAB-B strain (CCUG 48898) and compared them to 25 publically available genomes of globally diverse MAB strains. Genome-wide analyses of 27,598 core genome single nucleotide polymorphisms (SNPs) revealed that the two Brazilian derived CRM strains are nearly indistinguishable from one another and are more closely related to UK outbreak isolates infecting CF patients than to strains from the US, Malaysia or France. Comparative genomic analyses of six closely related outbreak strains revealed geographic-specific large-scale insertion/deletion variation that corresponds to bacteriophage insertions and recombination hotspots. Our study integrates new genome sequence data with existing genomic information to explore the global diversity of infectious M. abscessus isolates and to compare clinically relevant outbreak strains from different continents. PMID:24055961

  19. Characterization of novel microsatellite markers in Musa acuminata subsp. burmannicoides, var. Calcutta 4

    PubMed Central

    2010-01-01

    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 assisted selection for traits. PMID:20507605

  20. Characterization of nitrite degradation by Lactobacillus casei subsp. rhamnosus LCR 6013.

    PubMed

    Liu, Dong-mei; Wang, Pan; Zhang, Xin-yue; Xu, Xi-lin; Wu, Hui; Li, Li

    2014-01-01

    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(+). PMID:24755671

  1. Impact of Protein Shedding on Detection of Mycobacterium avium subsp. paratuberculosis by a Whole-Cell Immunoassay Incorporating Surface-Enhanced Raman Scattering?

    PubMed Central

    Yakes, Betsy Jean; Lipert, Robert J.; Bannantine, John P.; Porter, Marc D.

    2008-01-01

    The etiological agent of Johne's disease is Mycobacterium avium subsp. paratuberculosis. Controlling the spread of this disease is hindered by the lack of sensitive, selective, and rapid detection methods for M. avium subsp. paratuberculosis. By using a recently optimized sandwich immunoassay (B. J. Yakes, R. J. Lipert, J. P. Bannantine, and M. D. Porter, Clin. Vaccine Immunol. 15:227-234, 2008), which incorporates a new monoclonal antibody for the selective capture and labeling of M. avium subsp. paratuberculosis and surface-enhanced Raman scattering for sensitive readout, detection limits of ?630 and ?740 M. avium subsp. paratuberculosis cells/ml are achieved in phosphate-buffered saline and whole milk samples, respectively, after spiking with heat-treated M. avium subsp. paratuberculosis. Surprisingly, these detection limits are 3 orders of magnitude lower than expected based on theoretical predictions. Experiments designed to determine the origin of the improvement revealed that the major membrane protein targeted by the monoclonal antibody was present in the sample suspensions as shed protein. This finding indicates that the capture and labeling of shed protein function as a facile amplification strategy for lowering the limit of detection for M. avium subsp. paratuberculosis that may also be applicable to the design of a wide range of highly sensitive assays for other cells and viruses. PMID:18077615

  2. Detection of antibodies specific to Campylobacter fetus subsp. venerealis in the vaginal mucus of Nigerian breeding cows.

    PubMed

    Mshelia, Gideon D; Amin, Jibrilla D; Egwu, Godwin O; Yavari, Christine A; Murray, Richard D; Woldehiwet, Zerai

    2010-01-01

    The presence of bovine venereal campylobacteriosis in the Lake Chad Basin of Nigeria was investigated using an enzyme-linked immunosorbent assay (ELISA) for the detection of IgA antibodies specific to Campylobacter fetus subsp. venerealis in vaginal mucus (n = 66). IgA antibodies specific to C. fetus subsp. venerealis were detected in 7 (11%) vaginal mucus samples. All but one of the IgA-positive samples originated from cows belonging to herds with a history of abortion and infertility which suggested an association between antibody detection and poor herd fertility. It was concluded that bovine venereal campylobacteriosis is prevalent in the Lake Chad Basin of Nigeria and its contribution to reduced reproductive performance in cattle herds may be grossly underestimated in this part of the world. PMID:20857383

  3. Molecular and immunohistochemical diagnosis of Francisella noatunensis subsp. orientalis from formalin-fixed, paraffin-embedded tissues.

    PubMed

    Soto, Esteban; Illanes, Oscar; Hilchie, David; Morales, Juan A; Sunyakumthorn, Piyanate; Hawke, John P; Goodwin, Andrew E; Riggs, Allen; Yanong, Roy P; Pouder, Deborah B; Francis-Floyd, Ruth; Arauz, Maziel; Bogdanovic, Lewis; Castillo-Alcala, Fernanda

    2012-09-01

    Members of the genus Francisella (viz., F. noatunensis subsp. orientalis [Fno] and F. noatunensis subsp. noatunensis) have been described as causative agents of chronic granulomatous and pyogranulomatous lesions in wild and cultured fish species. In the present study, 68 archived formalin-fixed, paraffin-embedded (FFPE) tissues from several fish species, collected at different geographical locations from 2000 to 2011, were analyzed using a real-time polymerase chain reaction assay for the detection of the Fno intracellular growth loci C (iglC) gene and by immunohistochemistry for the demonstration of Fno antigens. The results revealed a high correlation between these 2 diagnostic techniques validating their use for the diagnosis of Fno infection in archived FFPE tissues and confirming the presence of Fno in fish species from the Cari y years of the present century. PMID:22786972

  4. First Complete Genome Sequence of a Salmonella enterica subsp. enterica Serovar Derby Strain Associated with Pork in France

    PubMed Central

    Hirchaud, Edouard; Rose, Valérie; Esnault, Emilie; Naquin, Delphine; Denis, Martine

    2015-01-01

    In France, Salmonella enterica subsp. enterica serovar Derby is one of the most often isolated serovars in pigs. Here, we describe the draft genome sequence of a strain isolated from a pig. This strain had the most frequent pulsed-field gel electrophoresis (PFGE) and antimicrobial patterns (S, SSU, T) usually observed in pig production in France. Those patterns have been also highlighted in human isolates. PMID:26227607

  5. First Complete Genome Sequence of a Salmonella enterica subsp. enterica Serovar Derby Strain Associated with Pork in France.

    PubMed

    Kérouanton, Annaëlle; Hirchaud, Edouard; Rose, Valérie; Esnault, Emilie; Naquin, Delphine; Denis, Martine

    2015-01-01

    In France, Salmonella enterica subsp. enterica serovar Derby is one of the most often isolated serovars in pigs. Here, we describe the draft genome sequence of a strain isolated from a pig. This strain had the most frequent pulsed-field gel electrophoresis (PFGE) and antimicrobial patterns (S, SSU, T) usually observed in pig production in France. Those patterns have been also highlighted in human isolates. PMID:26227607

  6. Genetic diversity of clinical Mycobacterium avium subsp. hominissuis and Mycobacterium intracellulare isolates causing pulmonary diseases recovered from different geographical regions.

    PubMed

    Ichikawa, Kazuya; van Ingen, Jakko; Koh, Won-Jung; Wagner, Dirk; Salfinger, Max; Inagaki, Takayuki; Uchiya, Kei-Ichi; Nakagawa, Taku; Ogawa, Kenji; Yamada, Kiyofumi; Yagi, Tetsuya

    2015-12-01

    Mycobacterium avium complex (MAC) infections are increasing annually in many countries. MAC strains are the most common nontuberculous mycobacterial pathogens isolated from respiratory samples and predominantly consist of two species, Mycobacterium avium and Mycobacterium intracellulare. The aim of this study was to analyze the molecular epidemiology and genetic backgrounds of clinical MAC isolates collected from The Netherlands, Germany, United States, Korea and Japan. Variable numbers of tandem repeats (VNTR) analysis was used to examine the genetic relatedness of clinical isolates of M. avium subsp. hominissuis (n=261) and M. intracellulare (n=116). Minimum spanning tree and unweighted pair group method using arithmetic averages analyses based on the VNTR data indicated that M. avium subsp. hominissuis isolates from Japan shared a high degree of genetic relatedness with Korean isolates, but not with isolates from Europe or the United States, whereas M. intracellulare isolates did not show any specific clustering by geographic origin. The findings from the present study indicate that strains of M. avium subsp. hominissuis, but not M. intracellulare, exhibit geographical differences in genetic diversity and imply that MAC strains may have different sources, routes of transmission and perhaps clinical manifestations. PMID:26435330

  7. Complete genome sequence of Polynucleobacter necessarius subsp. asymbioticus type strain (QLW-P1DMWA-1T)

    SciTech Connect

    Meincke, Linda; Copeland, A; Lapidus, Alla L.; Lucas, Susan; Berry, Kerrie W.; Glavina Del Rio, Tijana; Hammon, Nancy; Dalin, Eileen; Tice, Hope; Pitluck, Sam; Richardson, P M; Bruce, David; Goodwin, Lynne A.; Han, Cliff; Tapia, Roxanne; Detter, J. Chris; Schmutz, Jeremy; Brettin, Thomas S; Larimer, Frank W; Land, Miriam L; Hauser, Loren John; Kyrpides, Nikos C; Ivanova, N; Goker, Markus; Woyke, Tanja; Wu, Qinglong L.; Pockl, Matthias; Hahn, Martin W.; Klenk, Hans-Peter

    2012-01-01

    Polynucleobacter necessarius subsp. asymbioticus Hahn et al. 2009 is one of currently two subspecies of P. necessarius. While P. necessarius subsp. asymbioticus is a free-living bacterium, the closely related second subspecies, P. necessarius subsp. necessarius is an obligate endosymbiont living in the cytoplasm of freshwater ciliates of the genus Euplotes aediculatus. The two P. necessarius subspecies were the closest thus far reported phylogenetic neighbors that differ in their lifestyle as obligately free-living vs. obligate endosymbiontic, and they are the only members of the genus Polynucleobacter with completely sequenced genomes. The genome-sequenced strain represents a group of closely related strains not distinguishable by 16S rRNA, 16S-23S ITS or glnA sequences, which is persistent in the home habitat of the strain and frequently contributes > 10% of total bacterial numbers in water samples of the habitat. The 2,159,490 bp long chromosome with a total of 2,088 protein-coding and 48 RNA genes was sequenced as part of the DOE Joint Genome Institute Community Sequencing Program 2006.

  8. The Host Genotype and Environment Affect Strain Types of Bifidobacterium longum subsp. longum Inhabiting the Intestinal Tracts of Twins.

    PubMed

    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. PMID:25956768

  9. Exposure of Bifidobacterium longum subsp. infantis to Milk Oligosaccharides Increases Adhesion to Epithelial Cells and Induces a Substantial Transcriptional Response

    PubMed Central

    Kavanaugh, Devon W.; O’Callaghan, John; Buttó, Ludovica F.; Slattery, Helen; Lane, Jonathan; Clyne, Marguerite; Kane, Marian; Joshi, Lokesh; Hickey, Rita M.

    2013-01-01

    In this study, we tested the hypothesis that milk oligosaccharides may contribute not only to selective growth of bifidobacteria, but also to their specific adhesive ability. Human milk oligosaccharides (3?sialyllactose and 6?sialyllactose) and a commercial prebiotic (Beneo Orafti P95; oligofructose) were assayed for their ability to promote adhesion of Bifidobacterium longum subsp. infantis ATCC 15697 to HT-29 and Caco-2 human intestinal cells. Treatment with the commercial prebiotic or 3?sialyllactose did not enhance adhesion. However, treatment with 6?sialyllactose resulted in increased adhesion (4.7 fold), while treatment with a mixture of 3?- and 6?-sialyllactose substantially increased adhesion (9.8 fold) to HT-29 intestinal cells. Microarray analyses were subsequently employed to investigate the transcriptional response of B. longum subsp. infantis to the different oligosaccharide treatments. This data correlated strongly with the observed changes in adhesion to HT-29 cells. The combination of 3?- and 6?-sialyllactose resulted in the greatest response at the genetic level (both in diversity and magnitude) followed by 6?sialyllactose, and 3?sialyllactose alone. The microarray data was further validated by means of real-time PCR. The current findings suggest that the increased adherence phenotype of Bifidobacterium longum subsp. infantis resulting from exposure to milk oligosaccharides is multi-faceted, involving transcription factors, chaperone proteins, adhesion-related proteins, and a glycoside hydrolase. This study gives additional insight into the role of milk oligosaccharides within the human intestine and the molecular mechanisms underpinning host-microbe interactions. PMID:23805302

  10. Specific Discrimination of Three Pathogenic Salmonella enterica subsp. enterica Serotypes by carB-Based Oligonucleotide Microarray

    PubMed Central

    Shin, Hwa Hui; Hwang, Byeong Hee; Seo, Jeong Hyun

    2014-01-01

    It is important to rapidly and selectively detect and analyze pathogenic Salmonella enterica subsp. enterica in contaminated food to reduce the morbidity and mortality of Salmonella infection and to guarantee food safety. In the present work, we developed an oligonucleotide microarray containing duplicate specific capture probes based on the carB gene, which encodes the carbamoyl phosphate synthetase large subunit, as a competent biomarker evaluated by genetic analysis to selectively and efficiently detect and discriminate three S. enterica subsp. enterica serotypes: Choleraesuis, Enteritidis, and Typhimurium. Using the developed microarray system, three serotype targets were successfully analyzed in a range as low as 1.6 to 3.1 nM and were specifically discriminated from each other without nonspecific signals. In addition, the constructed microarray did not have cross-reactivity with other common pathogenic bacteria and even enabled the clear discrimination of the target Salmonella serotype from a bacterial mixture. Therefore, these results demonstrated that our novel carB-based oligonucleotide microarray can be used as an effective and specific detection system for S. enterica subsp. enterica serotypes. PMID:24185846

  11. Improved trehalose production from biodiesel waste using parent and osmotically sensitive mutant of Propionibacterium freudenreichii subsp. shermanii under aerobic conditions.

    PubMed

    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. PMID:22526328

  12. The Genome Sequence of the Tomato-Pathogenic Actinomycete Clavibacter michiganensis subsp. michiganensis NCPPB382 Reveals a Large Island Involved in Pathogenicity? †

    PubMed Central

    Gartemann, Karl-Heinz; Abt, Birte; Bekel, Thomas; Burger, Annette; Engemann, Jutta; Flügel, Monika; Gaigalat, Lars; Goesmann, Alexander; Gräfen, Ines; Kalinowski, Jörn; Kaup, Olaf; Kirchner, Oliver; Krause, Lutz; Linke, Burkhard; McHardy, Alice; Meyer, Folker; Pohle, Sandra; Rückert, Christian; Schneiker, Susanne; Zellermann, Eva-Maria; Pühler, Alfred; Eichenlaub, Rudolf; Kaiser, Olaf; Bartels, Daniela

    2008-01-01

    Clavibacter michiganensis subsp. michiganensis is a plant-pathogenic actinomycete that causes bacterial wilt and canker of tomato. The nucleotide sequence of the genome of strain NCPPB382 was determined. The chromosome is circular, consists of 3.298 Mb, and has a high G+C content (72.6%). Annotation revealed 3,080 putative protein-encoding sequences; only 26 pseudogenes were detected. Two rrn operons, 45 tRNAs, and three small stable RNA genes were found. The two circular plasmids, pCM1 (27.4 kbp) and pCM2 (70.0 kbp), which carry pathogenicity genes and thus are essential for virulence, have lower G+C contents (66.5 and 67.6%, respectively). In contrast to the genome of the closely related organism Clavibacter michiganensis subsp. sepedonicus, the genome of C. michiganensis subsp. michiganensis lacks complete insertion elements and transposons. The 129-kb chp/tomA region with a low G+C content near the chromosomal origin of replication was shown to be necessary for pathogenicity. This region contains numerous genes encoding proteins involved in uptake and metabolism of sugars and several serine proteases. There is evidence that single genes located in this region, especially genes encoding serine proteases, are required for efficient colonization of the host. Although C. michiganensis subsp. michiganensis grows mainly in the xylem of tomato plants, no evidence for pronounced genome reduction was found. C. michiganensis subsp. michiganensis seems to have as many transporters and regulators as typical soil-inhabiting bacteria. However, the apparent lack of a sulfate reduction pathway, which makes C. michiganensis subsp. michiganensis dependent on reduced sulfur compounds for growth, is probably the reason for the poor survival of C. michiganensis subsp. michiganensis in soil. PMID:18192381

  13. The genome of Aeromonas salmonicida subsp. salmonicida A449: insights into the evolution of a fish pathogen

    PubMed Central

    Reith, Michael E; Singh, Rama K; Curtis, Bruce; Boyd, Jessica M; Bouevitch, Anne; Kimball, Jennifer; Munholland, Janet; Murphy, Colleen; Sarty, Darren; Williams, Jason; Nash, John HE; Johnson, Stewart C; Brown, Laura L

    2008-01-01

    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 Aeromonas species. Conclusion Relative to the A. hydrophila ATCC 7966T genome, the A. salmonicida subsp. salmonicida genome has acquired multiple mobile genetic elements, undergone substantial rearrangement and developed a significant number of pseudogenes. These changes appear to be a consequence of adaptation to a specific host, salmonid fish, and provide insights into the mechanisms used by the bacterium for infection and avoidance of host defence systems. PMID:18801193

  14. Host Transcriptional Profiles and Immunopathologic Response following Mycobacterium avium subsp. paratuberculosis Infection in Mice

    PubMed Central

    Shin, Min-Kyoung; Park, Hongtae; Shin, Seung Won; Jung, Myunghwan; Lee, Su-Hyung; Kim, Dae-Yong; Yoo, Han Sang

    2015-01-01

    Paratuberculosis or Johne’s disease is a chronic granulomatous enteropathy in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP) infection. In the present study, we examined the host response to MAP infection in spleens of mice in order to investigate the host immunopathology accompanying host-pathogen interaction. Transcriptional profiles of the MAP-infected mice at 3 and 6 weeks p.i. showed severe histopathological changes, whereas those at 12 weeks p.i. displayed reduced lesion severity in the spleen and liver. MAP-infected mice at 3 and 6 weeks p.i. showed up-regulation of interferon-related genes, scavenger receptor, and complement components, suggesting an initial innate immune reaction, such as macrophage activation, bactericidal activity, and macrophage invasion of MAP. Concurrently, MAP-infected mice at 3 and 6 weeks p.i. were also suggested to express M2 macrophage phenotype with up-regulation of Mrc1, and Marco and down-regulation of MHC class II, Ccr7, and Irf5, and canonical pathways related to the T cell response including ICOS-ICOSL signaling in T helper cells, calcium-induced T lymphocyte apoptosis, and CD28 signaling in T helper cell. These results provide information which furthers the understanding of the immunopathologic response to MAP infection in mice, thereby providing insights valuable for research into the pathogenesis for MAP infection. PMID:26439498

  15. Mycobacterium avium subsp. avium found in raptors exposed to infected domestic fowl.

    PubMed

    Kriz, Petr; Kaevska, Marija; Bartejsova, Iva; Pavlik, Ivo

    2013-09-01

    We report a case of a falcon breeding facility, where raptors (both diurnal and nocturnal) were raised in contact with domestic fowl (Gallus gallus f. domesticus) infected by Mycobacterium avium subsp. avium. Fecal and environmental samples from 20 raptors and four common ravens (Corvus corax) were collected. Mycobacterium a. avium DNA was detected in feces of four raptors (bald eagle [Haliaeetus leucocephalus], eagle owl [Bubo bubo], barn owl [Tyto alba], and little owl [Athene noctua]) using triplex quantitative real-time PCR. As both the flock of domestic fowl and one of the infected raptors had the same origin (zoological collection), they might have had a common source of colonization/infection. However, the detection of M. a. avium in feces of three other raptors may point at transmission of the agent between the birds in the facility. Contact of raptors with domestic fowl infected by M. a. avium may pose a risk for transmission of the infection for them; however, raptors from the falcon breeding facility seemed to be relatively resistant to the infection. PMID:24283140

  16. Patterns of cytotype variation of Turnera sidoides subsp. pinnatifida (Turneraceae) in mountain ranges of central Argentina.

    PubMed

    Elías, Gabriela; Sartor, María; Solís Neffa, Viviana G

    2011-01-01

    Cytogeographical variability among 564 plants from 26 populations of Turnera sidoides subsp. pinnatifida in mountain ranges of central Argentina was analysed with meiotic chromosome counts and flow cytometry and is described at regional and local scales. Populations were primarily tetraploids (2n = 4x = 28), although diploid (2n = 2x = 14), hexaploid (2n = 2x = 42), and mixed populations of diploids and triploids (2n = 3x = 21) were also found. Diploids, triploids, and hexaploids were fewer in number and restricted to narrow areas, while tetraploids were the most common and geographically widespread cytotype. Diploids grew at higher altitudes and in colder and wet locations; tetraploids had the broadest ecological spectrum, while hexaploids occurred at the lowest altitudes and in drier conditions. The cytotypes were also spatially segregated at a microgeographical scale. Diploids grew in the piedmont, tetraploids were in the adjacent valley, and in the contact zone of both cytotypes, patches of diploids and triploids were found. At a regional scale, the distribution of the cytotypes may be governed by a combination of ecological and historical variables, while segregation in the contact zone may be independent of the selective environment because the cytotypes are unable to coexist as a result of reproductive exclusion. The role of triploids is also discussed. PMID:20437191

  17. Genetic transformation of intact Lactococcus lactis subsp. lactis by high-voltage electroporation

    SciTech Connect

    McIntyre, D.A.; Harlander, S.K. )

    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.

  18. Functional Characterization of a LOV-Histidine Kinase Photoreceptor from Xanthomonas citri subsp. citri.

    PubMed

    Kraiselburd, Ivana; Gutt, Alexander; Losi, Aba; Gärtner, Wolfgang; Orellano, Elena G

    2015-01-01

    The blue-light (BL) absorbing protein Xcc-LOV from Xanthomonas citri subsp. citri is composed of a LOV-domain, a histidine kinase (HK) and a response regulator. Spectroscopic characterization of Xcc-LOV identified intermediates and kinetics of the protein's photocycle. Measurements of steady state and time-resolved fluorescence allowed determination of quantum yields for triplet (?T  = 0.68 ± 0.03) and photoproduct formation (?390  = 0.46 ± 0.05). The lifetime for triplet decay was determined as ?T  = 2.4-2.8 ?s. Fluorescence of tryptophan and tyrosine residues was unchanged upon light-to-dark conversion, emphasizing the absence of significant conformational changes. Photochemistry was blocked upon cysteine C76 (C76S) mutation, causing a seven-fold longer lifetime of the triplet state (?T  = 16-18.5 ?s). Optoacoustic spectroscopy yielded the energy content of the triplet state. Interestingly, Xcc-LOV did not undergo the volume contraction reported for other LOV domains within the observation time window, although the back-conversion into the dark state was accompanied by a volume expansion. A radioactivity-based enzyme function assay revealed a larger HK activity in the lit than in the dark state. The C76S mutant showed a still lower enzyme function, indicating the dark state activity being corrupted by a remaining portion of the long-lived lit state. PMID:26172037

  19. The anti-infective activity of punicalagin against Salmonella enterica subsp. enterica serovar typhimurium in mice.

    PubMed

    Li, Guanghui; Feng, Yuqing; Xu, Yunfeng; Wu, Qian; Han, Qi'an; Liang, Xiujun; Yang, Baowei; Wang, Xin; Xia, Xiaodong

    2015-07-01

    Punicalagin, a major bioactive component of pomegranate peel, has been proven to have antioxidant, antiviral, anti-apoptosis, and hepatoprotective properties. The aim of this study was to investigate the anti-infective activity of punicalagin in a mouse model. C57BL/6 mice were initially challenged with Salmonella enterica subsp. enterica serovar typhimurium (S. typhimurium) and then treated with punicalagin. Food and water consumption and body weight were recorded daily. On day 8 post infection, the mice were sacrificed to examine pathogen counts in tissues, hematological parameters, cytokine levels, and histological changes. Compared to mice only infected with S. typhimurium, punicalagin-treated mice had more food consumption and less weight loss. A higher survival rate and lower counts of viable S. typhimurium in feces, liver, spleen, and kidney were found in the punicalagin-treated mice. The enzyme linked immunosorbent assay showed that the levels of IL-6, IL-10, and IFN-? in serum and the spleen and TNF-? in serum, the spleen and the liver were reduced by punicalagin. Moreover, more neutrophils and higher neutrophil-to-mononuclear cell ratios in the punicalagin-treated mice were observed. Histological examination showed that punicalagin protected cells in the liver and spleen from hemorrhagic necrosis. It is concluded that punicalagin has a beneficial effect against S. typhimurium infection in mice. The anti-infective properties, together with other nutritionally beneficial effects, make punicalagin a promising supplement in human food or animal feeds to prevent disease associated with S. typhimurium. PMID:26084785

  20. Red brome (Bromus rubens subsp. madritensis) in North America: Possible modes for early introductions, subsequent spread

    USGS Publications Warehouse

    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.

  1. Biochemical characterization of different genotypes of Paenibacillus larvae subsp. larvae, a honey bee bacterial pathogen.

    PubMed

    Neuendorf, Sandra; Hedtke, Kati; Tangen, Gerhard; Genersch, Elke

    2004-07-01

    Paenibacillus larvae subsp. larvae (P. l. larvae) is the aetiological agent of American foulbrood (AFB), the most virulent bacterial disease of honey bee brood worldwide. In many countries AFB is a notifiable disease since it is highly contagious, in most cases incurable and able to kill affected colonies. Genotyping of field isolates of P. l. larvae revealed at least four genotypes (AB, Ab, ab and alpha B) present in Germany which are genotypically different from the reference strain DSM 7030. Therefore, based on these data, five different genotypes of P. l. larvae are now identified with genotype AB standing out with a characteristic brown-orange and circled two-coloured colony morphology. Analysing the metabolic profiles of three German genotypes (AB, Ab and ab) as well as of the reference strain using the Biolog system, a characteristic biochemical fingerprint could be obtained for each strain. Cluster analysis showed that while genotypes Ab, ab and the reference strain DSM 7030 are rather similar, genotype AB is clearly different from the others. Analysis of all isolates for plasmid DNA revealed two different plasmids present only in isolates belonging to genotype AB. Therefore, genotype AB is remarkable in all aspects analysed so far. Future analysis will show whether or not these differences will expand to differences in virulence. PMID:15256579

  2. Studies on the biology of the crisamicin-producing organism Micromonospora purpureochromogenes subsp. celinoensis

    SciTech Connect

    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 /sup 13/C acetate feeding and /sup 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.

  3. Chemical composition, antioxidant and antibacterial properties of Bene (Pistacia atlantica subsp. mutica) hull essential oil.

    PubMed

    Rezaie, Mitra; Farhoosh, Reza; Sharif, Ali; Asili, Javad; Iranshahi, Mehrdad

    2015-10-01

    The tree Pistacia atlantica subsp. mutica, namely Bene, is widely distributed in Iranian mountains. Recent studies revealed that the oil of Bene was stable, even more stable than sesame oil, with antioxidant properties. This can give versatile applications for the oil. The volatile composition of this oil has not chemically been investigated so far. In this study, sixty three compounds were identified in the essential oil (EO) of Bene hull. The major components were determined to be ?-pinene (20.8 %), camphene (8.4 %), ?-myrcene (8.2 %) and limonene (8 %). Antioxidant activities of the essential oil from Bene hull were evaluated by using 2,2'- diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging, ferric reducing antioxidant power (FRAP), ?-carotene bleaching test, thiobarbituric acid reactive species (TBARS) and Rancimat assays. The Bene essential oil exhibited significant antioxidant activities in FRAP and TBARS assays as compared with positive controls. In addition, the oil was evaluated for its antimicrobial activity against both Gram positive (Staphylococcus aureus) and Gram negative (Escherichia coli) bacteria. It showed significant antibacterial activities against S. aureus and E. coli with minimum inhibitory concentration (MIC) values of 6 and 12.5 ?g/mL, respectively. PMID:26396430

  4. Dendroclimatic potential of plains cottonwood (Populus deltoides subsp. monilifera) from the Northern Great Plains, USA

    USGS Publications Warehouse

    Edmonson, Jesse; Friedman, Jonathan; Meko, David; Touchan, Ramzi; Scott, Julian; Edmonson, Alan

    2014-01-01

    A new 368-year tree-ring chronology (A.D. 1643–2010) has been developed in western North Dakota using plains cottonwood (Populus deltoides subsp. monilifera) growing on the relatively undisturbed floodplain of the Little Missouri River in the North Unit of Theodore Roosevelt National Park. We document many slow-growing living trees between 150–370 years old that contradict the common understanding that cottonwoods grow fast and die young. In this northern location, cottonwood produces distinct annual rings with dramatic interannual variability that strongly crossdate. The detrended tree-ring chronology is significantly positively correlated with local growing season precipitation and soil moisture conditions (r ?=? 0.69). This time series shows periods of prolonged low radial tree growth during the known droughts of the instrumental record (e.g. 1931–1939 and 1980–1981) and also during prehistory (e.g. 1816–1823 and 1856–1865) when other paleoclimate studies have documented droughts in this region. Tree rings of cottonwood will be a useful tool to help reconstruct climate, streamflow, and the floodplain history of the Little Missouri River and other northern river systems.

  5. LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis

    PubMed Central

    Trangoni, Marcos D.; Gioffré, Andrea K.; Cerón Cucchi, María E.; Caimi, Karina C.; Ruybal, Paula; Zumárraga, Martín J.; Cravero, Silvio L.

    2015-01-01

    In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries. PMID:26273282

  6. Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

    PubMed Central

    Johnston, Christopher D.; Bannantine, John P.; Govender, Rodney; Endersen, Lorraine; Pletzer, Daniel; Weingart, Helge; Coffey, Aidan; O'Mahony, Jim; Sleator, Roy D.

    2014-01-01

    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 MAP2121c can enhance the heterologous expression of the major membrane protein (MMP), analogous to the form in which it is 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, we previously engineered MAP3733c (encoding MptD) and show herein that MptD displays the tendency to associate with the cytoplasmic membrane boundary under confocal microscopy and the intracellularly accumulated protein selectively adheres to 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. PMID:25237653

  7. Transcriptional profile of Salmonella enterica subsp. enterica serovar Weltevreden during alfalfa sprout colonization

    PubMed Central

    Brankatschk, Kerstin; Kamber, Tim; Pothier, Joël F; Duffy, Brion; Smits, Theo H M

    2014-01-01

    Sprouted seeds represent a great risk for infection by human enteric pathogens because of favourable growth conditions for pathogens during their germination. The aim of this study was to identify mechanisms of interactions of Salmonella enterica subsp. enterica?Weltevreden with alfalfa sprouts. RNA-seq analysis of S. Weltevreden grown with sprouts in comparison with M9-glucose medium showed that among a total of 4158 annotated coding sequences, 177 genes (4.3%) and 345 genes (8.3%) were transcribed at higher levels with sprouts and in minimal medium respectively. Genes that were higher transcribed with sprouts are coding for proteins involved in mechanisms known to be important for attachment, motility and biofilm formation. Besides gene expression required for phenotypic adaption, genes involved in sulphate acquisition were higher transcribed, suggesting that the surface on alfalfa sprouts may be poor in sulphate. Genes encoding structural and effector proteins of Salmonella pathogenicity island 2, involved in survival within macrophages during infection of animal tissue, were higher transcribed with sprouts possibly as a response to environmental conditions. This study provides insight on additional mechanisms that may be important for pathogen interactions with sprouts. PMID:24308841

  8. Structural investigation of cell wall polysaccharides of Lactobacillus delbrueckii subsp. bulgaricus 17.

    PubMed

    Vinogradov, E; Sadovskaya, I; Cornelissen, A; van Sinderen, D

    2015-09-01

    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 (-[?-d-Glcp-(1-3)-]-4-?-l-Rhap2OAc-4-?-d-Glcp-[?-d-Galp-(1-3)]-4-?-Rhap-3-?-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-?-Galf-3-?-Galp-2-?-Galf-6-?-Galf-3-?-Galp-). PMID:26117227

  9. Bartonella clarridgeiae and Bartonella vinsonii subsp. berkhoffii exposure in captive wild canids in Brazil.

    PubMed

    Fleischman, D A; Chomel, B B; Kasten, R W; André, M R; Gonçalves, L R; Machado, R Z

    2015-02-01

    SUMMARY Wild canids are potential hosts for numerous species of Bartonella, yet little research has been done to quantify their infection rates in South America. We sought to investigate Bartonella seroprevalence in captive wild canids from 19 zoos in São Paulo and Mato Grosso states, Brazil. Blood samples were collected from 97 wild canids belonging to four different native species and three European wolves (Canis lupus). Indirect immunofluorescent antibody testing was performed to detect the presence of B. henselae, B. vinsonii subsp. berkhoffii, B. clarridgeiae, and B. rochalimae. Overall, Bartonella antibodies were detected in 11 of the canids, including five (12·8%) of 39 crab-eating foxes (Cerdocyon thous), three (11·1%) of 27 bush dogs (Speothos venaticus), two (8·7%) of 23 maned wolves (Chrysocyon brachyurus) and one (12·5%) of eight hoary foxes (Lycalopex vetulus), with titres ranging from 1:64 to 1:512. Knowing that many species of canids make excellent reservoir hosts for Bartonella, and that there is zoonotic potential for all Bartonella spp. tested for, it will be important to conduct further research in non-captive wild canids to gain an accurate understanding of Bartonella infection in free-ranging wild canids in South America. PMID:24892580

  10. From mouth to macrophage: mechanisms of innate immune subversion by Mycobacterium avium subsp. paratuberculosis

    PubMed Central

    2014-01-01

    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. PMID:24885748

  11. Oligosaccharide Binding Proteins from Bifidobacterium longum subsp. infantis Reveal a Preference for Host Glycans

    PubMed Central

    Garrido, Daniel; Kim, Jae Han; German, J. Bruce; Raybould, Helen E.; Mills, David A.

    2011-01-01

    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. PMID:21423604

  12. Biodistribution of a Promising Probiotic, Bifidobacterium longum subsp. longum Strain BBMN68, in the Rat Gut.

    PubMed

    Lv, Yang; Qiao, Xuewei; Zhao, Liang; Ren, Fazheng

    2015-06-01

    Bifidobacterium longum subsp. longum BBMN68, isolated from centenarians in Guangxi, China, has been proved to be a promising probiotic strain for its health benefits. In this study, the biodistribution of this strain in the rat gut was first investigated using the quantitative realtime PCR assay and propidium monoazide. Strain-specific primers were originally designed based on the BBMN68 genome sequence. Healthy rats were orally inoculated with either a single dose of BBMN68 (10(10) colony-forming units/kg), or with one dose per day for 7 days and bacterial concentrations were analyzed in detail from the intestinal contents and feces of four different gut locations, including stomach, small intestine, colon, and rectum. Results indicated that strain BBMN68 could overcome the rigors of passage through the upper gastrointestinal tract and transiently accumulate in the colon, even though survival in the stomach and small intestine was not high. A good level of BBMN8 could stay in vivo for 72 h following a 7-day oral administration, and a daily administration is suggested for a considerable and continuous population of BBMN68 to be maintained in the host intestine. PMID:25639716

  13. Molecular Clues To Understand the Aerotolerance Phenotype of Bifidobacterium animalis subsp. lactis

    PubMed Central

    Ruiz, Lorena; Gueimonde, Miguel; Ruas-Madiedo, Patricia; Ribbera, Angela; de los Reyes-Gavilán, Clara G.; Ventura, Marco; Sánchez, Borja

    2012-01-01

    Oxygen is one of the abiotic factors negatively affecting the survival of Bifidobacterium strains used as probiotics, mainly due to the induction of lethal oxidative damage. Aerobic conditions are present during the process of manufacture and storage of functional foods, and aerotolerance is a desired trait for bifidobacteria intended for use in industry. In the present study, the molecular response of Bifidobacterium animalis subsp. lactis IPLA4549 to aerobic conditions is presented. Molecular targets affected by oxygen were studied using two-dimensional electrophoresis (2DE) and quantitative reverse transcriptase (qRT) PCR. Globally, oxygen stress induced a shift in the glycolytic pathway toward the production of acetic acid with a concomitant increase in ATP formation. Several changes in the expression of genes coding for enzymes involved in redox reactions were detected, although the redox ratio remained unaltered. Interestingly, cells grown under aerobic conditions were characterized by higher activity of coproporphyrinogen III oxidase, which can directly detoxify molecular oxygen, and by higher NADH oxidase specific activity, which can oxidize NADH using hydrogen peroxide. In turn, this is in agreement with the glycolytic shift toward acetate production, in that more NADH molecules may be available due to the lower level of lactic acid formation. These findings further our ability to elucidate the mechanisms by which B. animalis copes with an oxygen-containing atmosphere. PMID:22101052

  14. Onchocerca dewittei japonica n. subsp., a common parasite from wild boar in Kyushu Island, Japan.

    PubMed

    Uni, S; Bain, O; Takaoka, H; Miyashita, M; Suzuki, Y

    2001-09-01

    We describe Onchocerca dewittei japonica n. subsp. from the Japanese wild boar, Sus scrofa leucomystax, in Oita, Kyushu Island, where all seven animals examined were found to be infected. This study began with efforts to identify the causative species in a recent case of zoonotic onchocerciasis. Compared with Onchocerca dewittei dewittei from Sus scrofa jubatus in Malaysia, which was reexamined here, our new subspecies has much greater space between the ridges on the females. In addition, its microfilariae (from uteri) are shorter (192-210 microns compared with 228-247 microns), and only the posterior third of the microfilarial body is coiled, instead of the entire body. The Onchocerca species parasitic in suids (these two subspecies and O. ramachandrini from the warthog in the Ethiopian region) form a group sharing several characters. Among the most unusual characters are the body swellings (a specialized apparatus for mating, known in only a few other genera). In addition, longitudinal cuticular crests were found on males of both subspecies from wild boar and on females of O. ramachandrini. PMID:11584751

  15. Phenolic compounds from Merremia umbellata subsp. orientalis and their allelopathic effects on Arabidopsis seed germination.

    PubMed

    Yan, Jian; Bi, Hai-Hong; Liu, Yong-Zhu; Zhang, Mei; Zhou, Zhong-Yu; Tan, Jian-Wen

    2010-11-01

    A bioassay-directed phytochemical study was carried out to investigate potential allelochemicals of the invasive plant Merremia umbellata subsp. orientalis (Hall. f.). Eight phenolic compounds, including a salicylic acid (SA)-derived new natural product, SA 2-O-?-D-(3',6'-dicaffeoyl)-glucopyranoside (1), and seven known ones 2-8 were isolated and identified from two bioactive sub-fractions of the acetone extract of this plant. The structure of new compound 1 was established by spectral and chemical methods. The potential allelopathic effects of these compounds at 0.5 and 1.0 mM concentrations on the germination of Arabidopsis seeds were tested. Results showed that 2 remarkably inhibited seed germination at concentrations as low as 0.5 mM. Compound 3 only moderately inhibited seed germination at 0.5 mM, but displayed strong inhibitory bioactivity at 1.0 mM concentration. Compounds 4 and 5 showed only slight inhibitory bioactivity at 1.0 mM, while the other compounds showed no obvious inhibitory effects. PMID:21076390

  16. Genome sequence of the enterobacterial phytopathogen Erwinia carotovora subsp. atroseptica and characterization of virulence factors

    PubMed Central

    Bell, K. S.; Sebaihia, M.; Pritchard, L.; Holden, M. T. G.; Hyman, L. J.; Holeva, M. C.; Thomson, N. R.; Bentley, S. D.; Churcher, L. J. C.; Mungall, K.; Atkin, R.; Bason, N.; Brooks, K.; Chillingworth, T.; Clark, K.; Doggett, J.; Fraser, A.; Hance, Z.; Hauser, H.; Jagels, K.; Moule, S.; Norbertczak, H.; Ormond, D.; Price, C.; Quail, M. A.; Sanders, M.; Walker, D.; Whitehead, S.; Salmond, G. P. C.; Birch, P. R. J.; Parkhill, J.; Toth, I. K.

    2004-01-01

    The bacterial family Enterobacteriaceae is notable for its well studied human pathogens, including Salmonella, Yersinia, Shigella, and Escherichia spp. However, it also contains several plant pathogens. We report the genome sequence of a plant pathogenic enterobacterium, Erwinia carotovora subsp. atroseptica (Eca) strain SCRI1043, the causative agent of soft rot and blackleg potato diseases. Approximately 33% of Eca genes are not shared with sequenced enterobacterial human pathogens, including some predicted to facilitate unexpected metabolic traits, such as nitrogen fixation and opine catabolism. This proportion of genes also contains an overrepresentation of pathogenicity determinants, including possible horizontally acquired gene clusters for putative type IV secretion and polyketide phytotoxin synthesis. To investigate whether these gene clusters play a role in the disease process, an arrayed set of insertional mutants was generated, and mutations were identified. Plant bioassays showed that these mutants were significantly reduced in virulence, demonstrating both the presence of novel pathogenicity determinants in Eca, and the impact of functional genomics in expanding our understanding of phytopathogenicity in the Enterobacteriaceae. PMID:15263089

  17. The investigation of some bioactive compounds and antioxidant properties of hawthorn (Crataegus monogyna subsp. monogyna Jacq)

    PubMed Central

    Keser, Serhat; Celik, Sait; Turkoglu, Semra; Yilmaz, Ökkes; Turkoglu, Ismail

    2014-01-01

    Aim: 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. Materials and Methods: For this purpose, antioxidant activities of extracts were assessed on DPPH•, ABTS•+, superoxide scavenging, reducing power and ferrous metal chelating activity assays and phenolic content of extracts was determined by Folin—Cioacalteu’s reagent. Results: The flavonoids including rutin, apigenin, myricetin, quercetin, naringenin and kaempferol, were identified by high-performance liquid chromatography in the hawthorn extract. Conclusion: It was observed the aqueous and ethanol extracts of Crataegus monogyna subsp. monogyna fruits showed the highest activity in reducing power and metal chelating activity assays. In addition, it was determined that the aqueous flower extract showed higher flavonoid content than aqueous leaves extract. The antioxidant and pharmacological effects of hawthorn have mainly been attributed to the polyphenolic contents. PMID:26401347

  18. DNA Macroarray Profiling of Lactococcus lactis subsp. lactis IL1403 Gene Expression during Environmental Stresses†

    PubMed Central

    Xie, Yi; Chou, Lan-szu; Cutler, Adele; Weimer, Bart

    2004-01-01

    This report describes the use of an oligonucleotide macroarray to profile the expression of 375 genes in Lactococcus lactis subsp. lactis IL1403 during heat, acid, and osmotic stress. A set of known stress-associated genes in IL1403 was used as the internal control on the array. Every stress response was accurately detected using the macroarray, compared to data from previous reports. As a group, the expression patterns of the investigated metabolic genes were significantly altered by heat, acid, and osmotic stresses. Specifically, 13 to 18% of the investigated genes were differentially expressed in each of the environmental stress treatments. Interestingly, the methionine biosynthesis pathway genes (metA-metB1 and metB2-cysK) were induced during heat shock, but methionine utilization genes, such as metK, were induced during acid stress. These data provide a possible explanation for the differences between acid tolerance mechanisms of L. lactis strains IL1403 and MG1363 reported previously. Several groups of transcriptional responses were common among the stress treatments, such as repression of peptide transporter genes, including the opt operon (also known as dpp) and dtpT. Reduction of peptide transport due to environmental stress will have important implications in the cheese ripening process. Although stress responses in lactococci were extensively studied during the last decade, additional information about this bacterium was gained from the use of this metabolic array. PMID:15528540

  19. Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication.

    PubMed

    Kaldestad, Marte; Haugland, Gyri T; Rønneseth, Anita; Wergeland, Heidrun I; Samuelsen, Ole Bent

    2014-02-01

    The granuloma disease caused by Francisella noatunensis subsp. noatunensis in farmed Atlantic cod has not been successfully treated by use of antibacterials, even when antibacterial resistance testing indicates a sufficient effect. The reason for this treatment failure may be the intracellular existence of the bacteria within immune cells, mainly macrophages. To investigate the effect of antibacterials on intracellular Francisella replication, we established a protocol for the detection of drugs within Atlantic cod immune cells using high-performance liquid chromatography (HPLC). When the uptake and intracellular concentrations of oxolinic acid and flumequine were analysed in isolated adherent head kidney leucocytes (HKLs) by HPLC, we found that uptake was rapid and the intracellular concentrations reflected the extracellular exposure concentrations. To investigate the effect of the antibacterial compounds on intracellular bacterial replication, adherent HKLs experimentally infected with the bacteria were analysed using flow cytometry and intracellular labelling of bacteria by specific antibodies. We found that flumequine did not inhibit intracellular bacterial replication. Unexpectedly, the results indicated that the intracellularly effiacy of the drug was reduced. The HPLC method used proved to be highly applicable for accurate determination of intracellular drug concentrations. When combined with sensitive and specific flow cytometry analyses for identification and measurement of intracellular bacterial replication, we suggest that this approach can be very valuable for the design of antibacterial treatments of intracellular pathogens. PMID:24492050

  20. A traditional Sudanese fermented camel's milk product, Gariss, as a habitat of Streptococcus infantarius subsp. infantarius.

    PubMed

    Abdelgadir, Warda; Nielsen, Dennis S; Hamad, Siddig; Jakobsen, Mogens

    2008-10-31

    Samples of the traditional Sudanese fermented camel's milk product Gariss representing 9 different regions in Sudan were microbiologically characterized using an integrated approach including phenotypic and genotypic methods. Lactic acid bacteria [log(CFU/g)=7.76-8.66] and yeasts [log(CFU/g)=6.05-7.79] were detected in high numbers. A total of 180 LAB isolates were identified of which 123 isolates were found to belong to the Streptococcus bovis group. Thirteen representative isolates were characterized by PCR amplification and sequencing of the housekeeping genes rpoB and sodA and the streptococcal glucosyltransferase gene (gtf). All thirteen isolates were identified as Streptococcus infantarius subsp. infantarius, a potential human pathogen. The gene encoding the virulence determinant gtf was detected in 10 of the 13 tested strains. The same isolates were able to survive exposure to 0.3% (w/v) oxgall for 4 h and pH=2.7 for 1-4 h. Also Lactobacillus fermentum were detected in high numbers, whereas Enterococcus faecium and Lactobacillus helveticus were detected more occasionally. The yeast microflora in all Gariss samples consisted of Kluyveromyces marxianus and Issatchenkia orientalis with the former being predominant in 7 out of 9 samples. PMID:18774196

  1. Safety and mutagenicity evaluation of Vigiis 101 powder made from Lactobacillus paracasei subsp. paracasei NTU 101.

    PubMed

    Tseng, Wei-Ting; Shih, Tsung-Wei; Liu, Shing-Hwa; Pan, Tzu-Ming

    2015-03-01

    The aim of the present work was to assess the genotoxic activity and the potential for toxicity upon repeated dosing of "Vigiis 101" powder, a probiotic consisting of dried bacteria Lactobacillus paracasei subsp. paracasei NTU 101. Results of the Ames test in Salmonella typhimurium strains TA1537, TA98, TA100, TA102, and TA1535 showed that Vigiis 101 (?5 mg per plate) was not mutagenic. We used experiments on ICR mice to evaluate the genotoxicity of Vigiis 101. Compared to the control, high-dose Vigiis 101 administration (16.72 g per kg of body weight) did not cause significant changes either in the number of reticulocytes or in the percentage (occurrence) of micronucleated reticulocytes. A mammalian chromosomal aberration test showed that the number of Chinese hamster ovary cells with abnormal chromosomes was <4% after Vigiis 101 treatment (maximal concentration was 5 mg/ml). A 28-day oral toxicity assay in Wistar rats was performed to assess the no-observed-adverse-effect level of Vigiis 101. Compared to the control, high-dose Vigiis 101 administration (5000 mg/kg/day) had no effects on mortality and body weight and did not cause toxicopathological lesions. Taken together, these results show that Vigiis 101 has no significant mutagenic or toxic effects. PMID:25481278

  2. Propionic acid production in glycerol/glucose co-fermentation by Propionibacterium freudenreichii subsp. shermanii.

    PubMed

    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. PMID:23584412

  3. Characterization of a Mycobacterium avium subsp. avium Operon Associated with Virulence and Drug Detoxification

    PubMed Central

    Viale, Mariana Noelia; Imperiale, Belén; Gioffre, Andrea Karina; Colombatti Olivieri, María Alejandra; Moyano, Roberto Damián; Morcillo, Nora; Santangelo, María de la Paz; Davis, William; Romano, María Isabel

    2014-01-01

    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. PMID:24967408

  4. The type III protein secretion system contributes to Xanthomonas citri subsp. citri biofilm formation

    PubMed Central

    2014-01-01

    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. PMID:24742141

  5. LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis.

    PubMed

    Trangoni, Marcos D; Gioffré, Andrea K; Cerón Cucchi, María E; Caimi, Karina C; Ruybal, Paula; Zumárraga, Martín J; Cravero, Silvio L

    2015-06-01

    In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR Green(TM) allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries. PMID:26273282

  6. Antiphagocytic Function of an IgG Glycosyl Hydrolase from Streptococcus equi subsp. equi and Its Use as a Vaccine Component

    PubMed Central

    Flock, Margareta; Frykberg, Lars; Sköld, Markus; Guss, Bengt

    2012-01-01

    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. PMID:22615244

  7. The Environment of “Mycobacterium avium subsp. hominissuis” Microaggregates Induces Synthesis of Small Proteins Associated with Efficient Infection of Respiratory Epithelial Cells

    PubMed Central

    Babrak, Lmar; Danelishvili, Lia; Rose, Sasha J.; Kornberg, Tiffany

    2014-01-01

    “Mycobacterium avium subsp. hominissuis” is an opportunistic environmental pathogen that causes respiratory illness in immunocompromised patients, such as those with cystic fibrosis as well as other chronic respiratory diseases. Currently, there is no efficient approach to prevent or treat M. avium subsp. hominissuis infection in the lungs. During initial colonization of the airways, M. avium subsp. hominissuis forms microaggregates composed of 3 to 20 bacteria on human respiratory epithelial cells, which provides an environment for phenotypic changes leading to efficient mucosal invasion in vitro and in vivo. DNA microarray analysis was employed to identify genes associated with the microaggregate phenotype. The gene encoding microaggregate-binding protein 1 (MBP-1) (MAV_3013) is highly expressed during microaggregate formation. When expressed in noninvasive Mycobacterium smegmatis, MBP-1 increased the ability of the bacteria to bind to HEp-2 epithelial cells. Using anti-MBP-1 immune serum, microaggregate binding to HEp-2 cells was significantly reduced. By far-Western blotting, and verified by coimmunoprecipitation, we observed that MBP-1 interacts with the host cytoskeletal protein vimentin. As visualized by confocal microscopy, microaggregates, as well as MBP-1, induced vimentin polymerization at the site of bacterium-host cell contact. Binding of microaggregates to HEp-2 cells was inhibited by treatment with an antivimentin antibody, suggesting that MBP-1 expression is important for M. avium subsp. hominissuis adherence to the host cell. MBP-1 immune serum significantly inhibited M. avium subsp. hominissuis infection throughout the respiratory tracts of mice. This study characterizes a pathogenic mechanism utilized by M. avium subsp. hominissuis to bind and invade the host respiratory epithelium, suggesting new potential targets for the development of antivirulence therapy. PMID:25422262

  8. Genetic structure of Mycobacterium avium subsp. paratuberculosis population in cattle herds in Quebec as revealed by using a combination of multilocus genomic analyses.

    PubMed

    Sohal, Jagdip Singh; Arsenault, Julie; Labrecque, Olivia; Fairbrother, Julie-Hélène; Roy, Jean-Philippe; Fecteau, Gilles; L'Homme, Yvan

    2014-08-01

    Mycobacterium avium subsp. paratuberculosis is the etiological agent of paratuberculosis, a granulomatous enteritis affecting a wide range of domestic and wild ruminants worldwide. A variety of molecular typing tools are used to distinguish M. avium subsp. paratuberculosis strains, contributing to a better understanding of M. avium subsp. paratuberculosis epidemiology. In the present study, PCR-based typing methods, including mycobacterial interspersed repetitive units/variable-number tandem repeats (MIRU-VNTR) and small sequence repeats (SSR) in addition to IS1311 PCR-restriction enzyme analysis (PCR-REA), were used to investigate the genetic heterogeneity of 200 M. avium subsp. paratuberculosis strains from dairy herds located in the province of Quebec, Canada. The majority of strains were of the "cattle type," or type II, although 3 strains were of the "bison type." A total of 38 genotypes, including a novel one, were identified using a combination of 17 genetic markers, which generated a Simpson's index of genetic diversity of 0.876. Additional analyses revealed no differences in genetic diversity between environmental and individual strains. Of note, a spatial and spatiotemporal cluster was evidenced regarding the distribution of one of the most common genotypes. The population had an overall homogeneous genetic structure, although a few strains stemmed out of the consensus cluster, including the bison-type strains. The genetic structure of M. avium subsp. paratuberculosis populations within most herds suggested intraherd dissemination and microevolution, although evidence of interherd contamination was also revealed. The level of genetic diversity obtained by combining MIRU-VNTR and SSR markers shows a promising avenue for molecular epidemiology investigations of M. avium subsp. paratuberculosis transmission patterns. PMID:24829229

  9. Effect of Turbulent-Flow Pasteurization on Survival of Mycobacterium avium subsp. paratuberculosis Added to Raw Milk

    PubMed Central

    Pearce, Lindsay E.; Truong, H. Tuan; Crawford, Robert A.; Yates, Gary F.; Cavaignac, Sonia; de Lisle, Geoffrey W.

    2001-01-01

    A pilot-scale pasteurizer operating under validated turbulent flow (Reynolds number, 11,050) was used to study the heat sensitivity of Mycobacterium avium subsp. paratuberculosis added to raw milk. The ATCC 19698 type strain, ATCC 43015 (Linda, human isolate), and three bovine isolates were heated in raw whole milk for 15 s at 63, 66, 69, and 72°C in duplicate trials. No strains survived at 72°C for 15 s; and only one strain survived at 69°C. Means of pooled D values (decimal reduction times) at 63 and 66°C were 15.0 ± 2.8 s (95% confidence interval) and 5.9 ± 0.7 s (95% confidence interval), respectively. The mean extrapolated D72°C was <2.03 s. This was equivalent to a >7 log10 kill at 72°C for 15 s (95% confidence interval). The mean Z value (degrees required for the decimal reduction time to traverse one log cycle) was 8.6°C. These five strains showed similar survival whether recovery was on Herrold's egg yolk medium containing mycobactin or by a radiometric culture method (BACTEC). Milk was inoculated with fresh fecal material from a high-level fecal shedder with clinical Johne's disease. After heating at 72°C for 15 s, the minimum M. avium subsp. paratuberculosis kill was >4 log10. Properly maintained and operated equipment should ensure the absence of viable M. avium subsp. paratuberculosis in retail milk and other pasteurized dairy products. An additional safeguard is the widespread commercial practice of pasteurizing 1.5 to 2° above 72°C. PMID:11525992

  10. Sensitive and Specific Enzyme-Linked Immunosorbent Assay for Detecting Serum Antibodies against Mycobacterium avium subsp. paratuberculosis in Fallow Deer

    PubMed Central

    Prieto, José M.; Balseiro, Ana; Casais, Rosa; Abendaño, Naiara; Fitzgerald, Liam E.; Garrido, Joseba M.; Juste, Ramon A.

    2014-01-01

    The enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies against Mycobacterium avium subsp. paratuberculosis in wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deer M. avium subsp. paratuberculosis isolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detect M. avium subsp. paratuberculosis antibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations. PMID:24872517

  11. Differential Expression of In Vivo and In Vitro Protein Profile of Outer Membrane of Acidovorax avenae Subsp. avenae

    PubMed Central

    Qiu, Hui; Li, Bin; Jabeen, Amara; Li, Liping; Liu, He; Kube, Michael; Xie, Guanlin; Wang, Yanli; 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. PMID:23166741

  12. Sensitive and specific enzyme-linked immunosorbent assay for detecting serum antibodies against Mycobacterium avium subsp. paratuberculosis in fallow deer.

    PubMed

    Prieto, José M; Balseiro, Ana; Casais, Rosa; Abendaño, Naiara; Fitzgerald, Liam E; Garrido, Joseba M; Juste, Ramon A; Alonso-Hearn, Marta

    2014-08-01

    The enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies against Mycobacterium avium subsp. paratuberculosis in wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deer M. avium subsp. paratuberculosis isolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detect M. avium subsp. paratuberculosis antibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations. PMID:24872517

  13. Effects of Elevated Atmospheric CO2 on Microbial Community Structure at the Plant-Soil Interface of Young Beech Trees (Fagus sylvatica L.) Grown at Two Sites with Contrasting Climatic Conditions.

    PubMed

    Gschwendtner, Silvia; Leberecht, Martin; Engel, Marion; Kublik, Susanne; Dannenmann, Michael; Polle, Andrea; Schloter, Michael

    2015-05-01

    Soil microbial community responses to elevated atmospheric CO2 concentrations (eCO2) occur mainly indirectly via CO2-induced plant growth stimulation leading to quantitative as well as qualitative changes in rhizodeposition and plant litter. In order to gain insight into short-term, site-specific effects of eCO2 on the microbial community structure at the plant-soil interface, young beech trees (Fagus sylvatica L.) from two opposing mountainous slopes with contrasting climatic conditions were incubated under ambient (360 ppm) CO2 concentrations in a greenhouse. One week before harvest, half of the trees were incubated for 2 days under eCO2 (1,100 ppm) conditions. Shifts in the microbial community structure in the adhering soil as well as in the root rhizosphere complex (RRC) were investigated via TRFLP and 454 pyrosequencing based on 16S ribosomal RNA (rRNA) genes. Multivariate analysis of the community profiles showed clear changes of microbial community structure between plants grown under ambient and elevated CO2 mainly in RRC. Both TRFLP and 454 pyrosequencing showed a significant decrease in the microbial diversity and evenness as a response of CO2 enrichment. While Alphaproteobacteria dominated by Rhizobiales decreased at eCO2, Betaproteobacteria, mainly Burkholderiales, remained unaffected. In contrast, Gammaproteobacteria and Deltaproteobacteria, predominated by Pseudomonadales and Myxococcales, respectively, increased at eCO2. Members of the order Actinomycetales increased, whereas within the phylum Acidobacteria subgroup Gp1 decreased, and the subgroups Gp4 and Gp6 increased under atmospheric CO2 enrichment. Moreover, Planctomycetes and Firmicutes, mainly members of Bacilli, increased under eCO2. Overall, the effect intensity of eCO2 on soil microbial communities was dependent on the distance to the roots. This effect was consistent for all trees under investigation; a site-specific effect of eCO2 in response to the origin of the trees was not observed. PMID:25370887

  14. Phylogenetic evidence suggests that strains of Aeromonas hydrophila subsp. dhakensis belong to the species Aeromonas aquariorum sp. nov.

    PubMed

    Martínez-Murcia, Antonio; Monera, Arturo; Alperi, Anabel; Figueras, Maria-José; Saavedra, Maria-José

    2009-01-01

    Three strains of Aeromonas hydrophila subspecies dhakensis, including the type strain, were subjected to phylogenetic analysis by sequencing gyrB, rpoD, and 16S rRNA genes and compared with all known Aeromonas species. The obtained gyrB and rpoD phylogenetic trees clearly suggested that these A. hydrophila subsp. dhakensis strains indeed belong to the species A. aquariorum. This finding may indicate that, at the time of "dhakensis" subspecies description, the strains were incorrectly identified as A. hydrophila by a polyphasic approach that included DNA-DNA hybridization. PMID:18839248

  15. Antibody responses elicited against the Treponema pallidum repeat proteins differ during infection with different isolates of Treponema pallidum subsp. pallidum.

    PubMed

    Leader, Brandon T; Hevner, Karin; Molini, Barbara J; Barrett, Lynn K; Van Voorhis, Wesley C; Lukehart, Sheila A

    2003-10-01

    Variation in the expression of the different Tpr proteins in the syphilis spirochete, Treponema pallidum subsp. pallidum, may have important implications in its ability to evade host immune detection and cause persistent infection. In the present study we examined the pattern of antibody responsiveness to different Tpr members during infection with three isolates of T. pallidum. There was variability in the specificities and temporal patterns of reactivity of the antibodies elicited against the individual Tpr proteins, suggesting that isolates may express different repertoires of Tpr proteins during infection. PMID:14500529

  16. Bacteriocinogenic Lactococcus lactis subsp. lactis DF04Mi isolated from goat milk: Evaluation of the probiotic potential

    PubMed Central

    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 probiotic potential. Lc. lactis DF4Mi was resistant to acidic pH and oxbile, presented co-aggregation with Listeria monocytogenes, and was not affected by several drugs from different generic groups, being sensitive to most tested antibiotics. These properties indicate that this Lc. lactis strain can be used for enhancement of dairy foods safety and quality, in combination with potential probiotic properties. PMID:25477942

  17. Bacillus thuringiensis subsp. sichuansis strain MC28 produces a novel crystal protein with activity against Culex quinquefasciatus larvae.

    PubMed

    Guan, Peng; Dai, Xiaojuan; Zhu, Jun; Li, Qiao; Li, Shuangcheng; Wang, Shiquan; Li, Ping; Zheng, Aiping

    2014-04-01

    The Bacillus thuringiensis subsp. sichuansis MC28 strain produces spherical parasporal crystals during sporulation and exhibits remarkable insecticidal activity against dipteran and lepidopteran pests. We characterized a novel cry gene (cry69Aa1), which was found in the pMC95 plasmid of the MC28 strain. The cry69Aa1 gene was inserted into a shuttle vector (pSTK) and expressed in an acrystalliferous mutant B. thuringiensis HD73?. In this transformant, a large number of spherical parasporal crystals, which were toxic to Culex quinquefasciatus (Diptera), were formed. PMID:24185745

  18. Epidemiology of a Salmonella enterica subsp. Enterica serovar Typhimurium strain associated with a songbird outbreak.

    USGS Publications Warehouse

    Blehert, David S.; Hernandez, Sonia M.; Keel, Kevin; Sanchez, Susan; Trees, Eija; Peter Gerner-Smidt

    2012-01-01

    Salmonella enterica subsp. enterica serovar Typhimurium is responsible for the majority of salmonellosis cases worldwide. This Salmonella serovar is also responsible for die-offs in songbird populations. In 2009, there was an S. Typhimurium epizootic reported in pine siskins in the eastern United States. At the time, there was also a human outbreak with this serovar that was associated with contaminated peanuts. As peanuts are also used in wild-bird food, it was hypothesized that the pine siskin epizootic was related to this human outbreak. A comparison of songbird and human S. Typhimurium pulsed-field gel electrophoresis (PFGE) patterns revealed that the epizootic was attributed not to the peanut-associated strain but, rather, to a songbird strain first characterized from an American goldfinch in 1998. This same S. Typhimurium strain (PFGE type A3) was also identified in the PulseNet USA database, accounting for 137 of 77,941 total S. Typhimurium PFGE entries. A second molecular typing method, multiple-locus variable-number tandem-repeat analysis (MLVA), confirmed that the same strain was responsible for the pine siskin epizootic in the eastern United States but was distinct from a genetically related strain isolated from pine siskins in Minnesota. The pine siskin A3 strain was first encountered in May 2008 in an American goldfinch and later in a northern cardinal at the start of the pine siskin epizootic. MLVA also confirmed the clonal nature of S. Typhimurium in songbirds and established that the pine siskin epizootic strain was unique to the finch family. For 2009, the distribution of PFGE type A3 in passerines and humans mirrored the highest population density of pine siskins for the East Coast.

  19. Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a

    PubMed Central

    Malheiros, Patrícia S.; Sant’Anna, Voltaire; Todorov, Svetoslav D.; Franco, Bernadette D.G.M.

    2015-01-01

    Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 24 factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R 2 = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L?1 and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL?1) occurred in the MRS broth supplemented with 5.5 g L?1 glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL?1. PMID:26413066

  20. Maternal sex effects and inbreeding depression under varied environmental conditions in gynodioecious Fragaria vesca subsp. bracteata

    PubMed Central

    Dalton, Rebecca M.; Koski, Matthew H.; Ashman, Tia-Lynn

    2013-01-01

    Background and Aims Gynodioecy (coexistence of females and hermaphrodites) is a sexual system that occurs in numerous flowering plant lineages. Thus, understanding the features that affect its maintenance has wide importance. Models predict that females must have a seed fitness advantage over hermaphrodites, and this may be achieved via seed quality or quantity. Females in a population of Fragaria vesca subsp. bracteata, a long-lived gynodioecious perennial, do not demonstrate a seed quantity advantage, so this study explored whether females produced better quality seed via maternal sex effects or avoidance of inbreeding depression (IBD). Methods Families of selfed and outcrossed seed were created using hermaphrodite mothers and families of outcrossed seed were created using female mothers. The effects of these pollination treatments were assessed under benign conditions early in life and under varied conditions later in life. To test for an effect of maternal sex, fitness components and traits associated with acclimation to variable environments of progeny of outbred hermaphrodites and females were compared. To test for expression of IBD, fitness parameters between inbred and outbred progeny of hermaphrodites were compared. Key Results Offspring of females were more likely to germinate in benign conditions and survive in harsh resource environments than outbred progeny of hermaphrodites. IBD was low across most life stages, and both the effect of maternal sex on progeny quality and the expression of IBD depended on both maternal family and resource condition of the progeny. Conclusions The effect of maternal sex and IBD on progeny quality depended on resource conditions, maternal lineage and progeny life stage. In conjunction with known lack of differences in seed quantity, the quality advantages and IBD observed here are still unlikely to be sufficient for maintenance of gynodioecy under nuclear inheritance of male sterility. PMID:23723257

  1. Plant Pathogenic Microbial Communication Affected by Elevated Temperature in Pectobacterium carotovorum subsp. carotovorum.

    PubMed

    Saha, N D; Chaudhary, A; Singh, S D; Singh, D; Walia, S; Das, T K

    2015-11-01

    Gram-negative plant pathogenic bacteria regulate specific gene expression in a population density-dependent manner by sensing level of Acyl-Homoserine Lactone (HSL) molecules which they produce and liberate to the environment, called Quorum Sensing (QS). The production of virulence factors (extracellular enzyme viz. cellulase, pectinase, etc.) in Pectobacterium carotovorum subsp. carotovorum (Pcc) is under strong regulation of QS. The QS signal molecule, N-(3-oxohexanoyl)-L-Homoserine Lactone (OHHL) was found as the central regulatory system for the virulence factor production in Pcc and is also under strict regulation of external environmental temperature. Under seven different incubation temperatures (24, 26, 28, 30, 33, 35, and 37 °C) in laboratory condition, highest amount of OHHL (804 violacein unit) and highest (79 %) Disease Severity Index (DSI) were measured at 33 °C. The OHHL production kinetics showed accumulation of highest concentration of OHHL at late log phase of the growth but diminution in the concentration occurred during stationary phase onwards to death phase. At higher temperature (35 and 37 °C) exposure, OHHL was not at detectable range. The effect of temperature on virulence factor production is the concomitant effect of HSL production and degradation which justifies less disease severity index in cross-inoculated tomato fruits incubated at 35 and 37 °C. The nondetection of the OHHL in the elevated temperature may because of degradation as these signal molecules are quite sensitive and prone to get degraded under different physical factors. This result provides the rationale behind the highest disease severity up to certain elevated temperature and leaves opportunities for investigation on mutation, co-evolution of superior plant pathogen with more stable HSL signals-mediated pathogenesis under global warming context. PMID:26271295

  2. Identification and Phytotoxicity Assessment of Phenolic Compounds in Chrysanthemoides monilifera subsp. monilifera (Boneseed)

    PubMed Central

    Al Harun, Md Abdullah Yousuf; Johnson, Joshua; Uddin, Md Nazim; Robinson, Randall W.

    2015-01-01

    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 quantity of free proline in the target species also suggests that the plant was in a stressed condition due to litter allelopathy. These findings are important for better understanding the invasive potential of boneseed and in devising control strategies. PMID:26465595

  3. Detection of mycobacterium avium subsp. paratuberculosis in cheeses from small ruminants in Tuscany.

    PubMed

    Galiero, Alessia; Fratini, Filippo; Mataragka, Antonia; Turchi, Barbara; Nuvoloni, Roberta; Ikonomopoulos, John; Cerri, Domenico

    2016-01-18

    Paratuberculosis is an infectious disease which affects mainly domestic and wild ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Map has been associated with human diseases like Crohn disease, type-1 diabetes, sarcoidosis, multiple sclerosis and Hashimoto's thyroiditis. The aim of this study was to determine the level of Map positivity of cheeses produced in Tuscany (Italy) as an indication of human exposure to the specific pathogen. Sampling was focused on artisanal cheeses produced without commercial starter culture from raw sheep or goat milk, on small-scale farms. Samples were tested by quantitative PCR (qPCR) and culture. Map DNA was detected in 4/7 (57.14%) goat, and in 14/25 (56%) sheep cheeses by qPCR, whereas cultivation produced a positive result in only one case. This corresponded to a goat cheese that had also reacted positively by qPCR and yielded a viable Type S (sheep) strain of Map. The Map load of the tested samples based on qPCR ranged from 6×10 to 1.8×10(4)Map cells/g of cheese. The results indicate on average 56.57% and 66.6% positivity of cheese samples and farms, respectively. Hence, the type of cheeses that were analyzed within the context of this study seem to constitute a considerable source of human exposure to Map; although the question remains of whether the Map cells were present in a viable form, since positive results were almost exclusively recorded by qPCR. PMID:26555160

  4. Quantitative real-time PCR and culture examination of Mycobacterium avium subsp. paratuberculosis at farm level.

    PubMed

    Hanifian, Shahram; Khani, Sajjad; Barzegari, Abolfazl; Shayegh, Jalal

    2013-02-22

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease in ruminants and may contribute to Crohn's disease in humans. The aim of this study was to determine the occurrence and quantity of MAP in cattle feces and milk in the Iranian context. In addition, we evaluated the effect of cattle age as well as farming system as risk factors contributing to MAP load. For this, a total sample of 373 consisting of 150 cattle feces (CF), 150 individual cow's milk (ICM), as well as 73 bulk-tank milk (BTM) was collected randomly and regardless of the cattle health status. The samples were assayed using F57 quantitative real-time PCR (qPCR) and culture method. According to the results of qPCR which was found ? 10 times more sensitive than culture assay, MAP was detected in 68.66% (103/150) of the CF, 12% (18/150) of the ICM and 52.05% (38/73) of the BTM samples. In contrast to the previous reports, the quantity of MAP in the BTM (2.03-5.97 log cfu/50 ml) was statistically (p<0.01) higher than the ICM (0.90-1.97 log cfu/50 ml). Data suggested a direct relation (p<0.01) between the cattle age and the quantity of MAP in the CF samples, while the relation was not statistically significant (p>0.05) for the ICM. In addition, MAP load in the BTM samples obtained from traditional farms was significantly (p<0.01) higher than that of the industrial ones, while the differences in CF and ICM was not significant (p>0.05). PMID:22999522

  5. Volatile Emissions from Mycobacterium avium subsp. paratuberculosis Mirror Bacterial Growth and Enable Distinction of Different Strains

    PubMed Central

    Trefz, Phillip; Koehler, Heike; Klepik, Klaus; Moebius, Petra; Reinhold, Petra; Schubert, Jochen K.; Miekisch, Wolfram

    2013-01-01

    Control of paratuberculosis in livestock is hampered by the low sensitivity of established direct and indirect diagnostic methods. Like other bacteria, Mycobacterium avium subsp. paratuberculosis (MAP) emits volatile organic compounds (VOCs). Differences of VOC patterns in breath and feces of infected and not infected animals were described in first pilot experiments but detailed information on potential marker substances is missing. This study was intended to look for characteristic volatile substances in the headspace of cultures of different MAP strains and to find out how the emission of VOCs was affected by density of bacterial growth. One laboratory adapted and four field strains, three of MAP C-type and one MAP S-type were cultivated on Herrold’s egg yolk medium in dilutions of 10-0, 10-2, 10-4 and 10-6. Volatile substances were pre-concentrated from the headspace over the MAP cultures by means of Solid Phase Micro Extraction (SPME), thermally desorbed from the SPME fibers and separated and identified by means of GC-MS. Out of the large number of compounds found in the headspace over MAP cultures, 34 volatile marker substances could be identified as potential biomarkers for growth and metabolic activity. All five MAP strains could clearly be distinguished from blank culture media by means of emission patterns based on these 34 substances. In addition, patterns of volatiles emitted by the reference strain were significantly different from the field strains. Headspace concentrations of 2-ethylfuran, 2-methylfuran, 3-methylfuran, 2-pentylfuran, ethyl acetate, 1-methyl-1-H-pyrrole and dimethyldisulfide varied with density of bacterial growth. Analysis of VOCs emitted from mycobacterial cultures can be used to identify bacterial growth and, in addition, to differentiate between different bacterial strains. VOC emission patterns may be used to approximate bacterial growth density. In a perspective volatile marker substances could be used to diagnose MAP infections in animals and to identify different bacterial strains and origins. PMID:24116177

  6. Essential oil composition and antibacterial activity of Origanum vulgare subsp. glandulosum Desf. at different phenological stages.

    PubMed

    Béjaoui, Afef; Chaabane, Hédia; Jemli, Maroua; Boulila, Abdennacer; Boussaid, Mohamed

    2013-12-01

    Variation in the quantity and quality of the essential oil (EO) of wild population of Origanum vulgare at different phenological stages, including vegetative, late vegetative, and flowering set, is reported. The oils of air-dried samples were obtained by hydrodistillation. The yield of oils (w/w%) at different stages were in the order of late vegetative (2.0%), early vegetative (1.7%), and flowering (0.6%) set. The oils were analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). In total, 36, 33, and 16 components were identified and quantified in vegetative, late vegetative, and flowering set, representing 94.47%, 95.91%, and 99.62% of the oil, respectively. Carvacrol was the major compound in all samples. The ranges of major constituents were as follows: carvacrol (61.08-83.37%), p-cymene (3.02-9.87%), and ?-terpinene (4.13-6.34%). Antibacterial activity of the oils was tested against three Gram-positive and two Gram-negative bacteria by the disc diffusion method and determining their diameter of inhibition and the minimum inhibitory concentration (MIC) values. The inhibition zones and MIC values for bacterial strains, which were sensitive to the EO of O. vulgare subsp. glandulosum, were in the range of 9-36 mm and 125-600 ?g/mL, respectively. The oils of various phenological stages showed high activity against all tested bacteria, of which Bacillus subtilis was the most sensitive and resistant strain, respectively. Thus, they represent an inexpensive source of natural antibacterial substances that exhibited potential for use in pathogenic systems. PMID:24320986

  7. Discovery of novel small molecule modulators of Clavibacter michiganensis subsp. michiganensis.

    PubMed

    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 ?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 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. PMID:26539169

  8. Predictor variables for and complications associated with Streptococcus equi subsp equi infection in horses.

    PubMed

    Duffee, Lauren R; Stefanovski, Darko; Boston, Raymond C; Boyle, Ashley G

    2015-11-15

    Objective-To evaluate predictor variables for and complications associated with Streptococcus equi subsp equi infection (strangles) in horses. Design-Retrospective case-control study. Animals-108 horses with strangles (cases) and 215 horses without strangles (controls). Procedures-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. Results-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. Conclusions and Clinical Relevance-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. PMID:26517620

  9. Characterization of a Novel Plasmid, pMAH135, from Mycobacterium avium Subsp. hominissuis

    PubMed Central

    Uchiya, Kei-ichi; Takahashi, Hiroyasu; Nakagawa, Taku; Yagi, Tetsuya; Moriyama, Makoto; Inagaki, Takayuki; Ichikawa, Kazuya; Nikai, Toshiaki; Ogawa, Kenji

    2015-01-01

    Mycobacterium avium complex (MAC) causes mainly two types of disease. The first is disseminated disease in immunocompromised hosts, such as individuals infected by human immunodeficiency virus (HIV). The second is pulmonary disease in individuals without systemic immunosuppression, and the incidence of this type is increasing worldwide. M. avium subsp. hominissuis, a component of MAC, causes infection in pigs as well as in humans. Many aspects of the different modes of M. avium infection and its host specificity remain unclear. Here, we report the characteristics and complete sequence of a novel plasmid, designated pMAH135, derived from M. avium strain TH135 in an HIV-negative patient with pulmonary MAC disease. The pMAH135 plasmid consists of 194,711 nucleotides with an average G + C content of 66.5% and encodes 164 coding sequences (CDSs). This plasmid was unique in terms of its homology to other mycobacterial plasmids. Interestingly, it contains CDSs with sequence homology to mycobactin biosynthesis proteins and type VII secretion system-related proteins, which are involved in the pathogenicity of mycobacteria. It also contains putative conserved domains of the multidrug efflux transporter. Screening of isolates from humans and pigs for genes located on pMAH135 revealed that the detection rate of these genes was higher in clinical isolates from pulmonary MAC disease patients than in those from HIV-positive patients, whereas the genes were almost entirely absent in isolates from pigs. Moreover, variable number tandem repeats typing analysis showed that isolates carrying pMAH135 genes are grouped in a specific cluster. Collectively, the pMAH135 plasmid contains genes associated with M. avium’s pathogenicity and resistance to antimicrobial agents. The results of this study suggest that pMAH135 influence not only the pathological manifestations of MAC disease, but also the host specificity of MAC infection. PMID:25671431

  10. Generation and screening of a comprehensive Mycobacterium avium subsp. paratuberculosis transposon mutant bank

    PubMed Central

    Rathnaiah, Govardhan; Lamont, Elise A.; Harris, N. Beth; Fenton, Robert J.; Zinniel, Denise K.; Liu, Xiaofei; Sotos, Josh; Feng, Zhengyu; Livneh-Kol, Ayala; Shpigel, Nahum Y.; Czuprynski, Charles J.; Sreevatsan, Srinand; Barletta, Raúl G.

    2014-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's Disease in ruminants. This enteritis has significant economic impact and worldwide distribution. Vaccination is one of the most cost effective infectious disease control measures. Unfortunately, current vaccines reduce clinical disease and shedding, but are of limited efficacy and do not provide long-term protective immunity. Several strategies have been followed to mine the MAP genome for virulence determinants that could be applied to vaccine and diagnostic assay development. In this study, a comprehensive mutant bank of 13,536 MAP K-10 Tn5367 mutants (P > 95%) was constructed and screened in vitro for phenotypes related to virulence. This strategy was designated to maximize identification of genes important to MAP pathogenesis without relying on studies of other mycobacterial species that may not translate into similar effects in MAP. This bank was screened for mutants with colony morphology alterations, susceptibility to D-cycloserine, impairment in siderophore production or secretion, reduced cell association, and decreased biofilm and clump formation. Mutants with interesting phenotypes were analyzed by PCR, Southern blotting and DNA sequencing to determine transposon insertion sites. These insertion sites mapped upstream from the MAP1152-MAP1156 cluster, internal to either the Mod operon gene MAP1566 or within the coding sequence of lsr2, and several intergenic regions. Growth curves in broth cultures, invasion assays and kinetics of survival and replication in primary bovine macrophages were also determined. The ability of vectors carrying Tn5370 to generate stable MAP mutants was also investigated. PMID:25360421

  11. Intracellular Trafficking of AIP56, an NF-?B-Cleaving Toxin from Photobacterium damselae subsp. piscicida

    PubMed Central

    Pereira, Liliana M. G.; Pinto, Rute D.; Silva, Daniela S.; Moreira, Ana R.; Beitzinger, Christoph; Oliveira, Pedro; Sampaio, Paula; Benz, Roland; Azevedo, Jorge E.; dos Santos, Nuno M. S.

    2014-01-01

    AIP56 (apoptosis-inducing protein of 56 kDa) is a metalloprotease AB toxin secreted by Photobacterium damselae subsp. piscicida that acts by cleaving NF-?B. During infection, AIP56 spreads systemically and depletes phagocytes by postapoptotic secondary necrosis, impairing the host phagocytic defense and contributing to the genesis of infection-associated necrotic lesions. Here we show that mouse bone marrow-derived macrophages (mBMDM) intoxicated by AIP56 undergo NF-?B p65 depletion and apoptosis. Similarly to what was reported for sea bass phagocytes, intoxication of mBMDM involves interaction of AIP56 C-terminal region with cell surface components, suggesting the existence of a conserved receptor. Biochemical approaches and confocal microscopy revealed that AIP56 undergoes clathrin-dependent endocytosis, reaches early endosomes, and follows the recycling pathway. Translocation of AIP56 into the cytosol requires endosome acidification, and an acidic pulse triggers translocation of cell surface-bound AIP56 into the cytosol. Accordingly, at acidic pH, AIP56 becomes more hydrophobic, interacting with artificial lipid bilayer membranes. Altogether, these data indicate that AIP56 is a short-trip toxin that reaches the cytosol using an acidic-pH-dependent mechanism, probably from early endosomes. Usually, for short-trip AB toxins, a minor pool reaches the cytosol by translocating from endosomes, whereas the rest is routed to lysosomes for degradation. Here we demonstrate that part of endocytosed AIP56 is recycled back and released extracellularly through a mechanism requiring phosphoinositide 3-kinase (PI3K) activity but independent of endosome acidification. So far, we have been unable to detect biological activity of recycled AIP56, thereby bringing into question its biological relevance as well as the importance of the recycling pathway. PMID:25287919

  12. Evaluation of viability Bifidobacterium animalis subsp. lactis LKM512 in dogs.

    PubMed

    Nakamura, A; Ohnishi, Y; Shirotori, K; Matsumoto, M

    2015-12-01

    In dogs, gastric acid is not neutralised even when a meal is present in the stomach. Moreover, dogs take longer to digest their meals than humans do. Accordingly, the most important characteristic of any probiotics considered for use in dogs is high acid tolerance. The probiotic strain Bifidobacterium animalis subsp. lactis LKM512 (hereafter referred to as LKM512) not only exhibits potent acid tolerance, but also has the ability to adhere to intestinal mucin. The aim of the present study was to explore the potential of LKM512 as a probiotic in dogs. Specifically, we investigated whether LKM512 can survive in the large intestine in dogs. LKM512 preparations containing 10(10) cfu were administered daily for 14 days in five dogs. Faeces were collected on the day before administration (day 0) as well as on days 7 and 14, and 7 days after administration was halted (day 21). The numbers of viable LKM512 present in faeces were determined by both culture-based techniques and molecular analysis. Changes in intestinal bacterial populations were analysed by 16S rRNA gene semiconductor sequencing using the Ion Torrent Personal Genome Machine (PGM). On days 7 and 14, the numbers of viable LKM512 that were detected in faeces by culture-based techniques and molecular analysis were greater than the original daily dosage. 16S rRNA gene sequence analysis using the PGM indicated that relative proportions of Bifidobacterium spp. and Bifidobacteriaceae were significantly higher after administration than before. The present study demonstrated that LKM512 can survive strong gastric acid, and proliferate in the large intestine of dogs. Therefore, LKM512 may be a useful canine probiotic. PMID:26322543

  13. Toxicological evaluation of the essential oil from Mentha longifolia L. subsp. capensis leaves in rats.

    PubMed

    Odeyemi, O O; Yakubu, M T; Masika, P J; Afolayan, A J

    2009-06-01

    The effects of the essential oil from the leaves of Mentha longifolia L. subsp. capensis on some biochemical parameters of Wistar rats were studied. The oil at 125, 250, 375, and 500 microL/kg of body weight reduced (P < .05) the red blood cells and lymphocytes with no definite pattern on the white blood cells and mean cell volume. The doses significantly increased the neutrophils, monocytes, large unstained cells, liver-body weight ratio, and serum concentrations of cholesterol, triglyceride, high-density lipoprotein-cholesterol, and inorganic phosphate but had no effect on the heart body weight ratio and serum low-density lipoprotein-cholesterol, Na(+), Ca(2+), Cl(-), K(+), creatinine, and uric acid. The oil at 500 microL/kg of body weight also increased the kidney-body weight ratio. In contrast, the oil reduced the serum urea and atherogenic index. The total and conjugated bilirubin, together with the total protein and albumin, in the serum increased only with oil at 125 microL/kg of body weight. The serum alkaline phosphatase activity also increased with no significant change in those of gamma-glutamyl transferase and alanine and aspartate aminotransferase. The results indicate dose- and parameter-specific effect of the essential oil. Although the essential oil from M. longifolia leaves may not predispose to atherosclerosis, it may increase the functional activity of the rat liver at the lowest dose investigated. Therefore, the essential oil from M. longifolia may not be completely "safe" at the doses investigated. PMID:19627219

  14. Metabolomic Profiling in Cattle Experimentally Infected with Mycobacterium avium subsp. paratuberculosis

    PubMed Central

    De Buck, Jeroen; Shaykhutdinov, Rustem; Barkema, Herman W.; Vogel, Hans J.

    2014-01-01

    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 was a sensitive method for detecting MAP infection much sooner than with current diagnostic methods, with individual metabolites significantly distinguishing infected from non-infected cattle. PMID:25372282

  15. Metabolic adaptation of Mycobacterium avium subsp. paratuberculosis to the gut environment.

    PubMed

    Weigoldt, Mathias; Meens, Jochen; Bange, Franz-Christoph; Pich, Andreas; Gerlach, Gerald F; Goethe, Ralph

    2013-02-01

    Knowledge on the proteome level about the adaptation of pathogenic mycobacteria to the environment in their natural hosts is limited. Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease, a chronic and incurable granulomatous enteritis of ruminants, and has been suggested to be a putative aetiological agent of Crohn's disease in humans. Using a comprehensive LC-MS-MS and 2D difference gel electrophoresis (DIGE) approach, we compared the protein profiles of clinical strains of MAP prepared from the gastrointestinal tract of diseased cows with the protein profiles of the same strains after they were grown in vitro. LC-MS-MS analyses revealed that the principal enzymes for the central carbon metabolic pathways, including glycolysis, gluconeogenesis, the tricaboxylic acid cycle and the pentose phosphate pathway, were present under both conditions. Moreover, a broad spectrum of enzymes for ?-oxidation of lipids, nine of which have been shown to be necessary for mycobacterial growth on cholesterol, were detected in vivo and in vitro. Using 2D-DIGE we found increased levels of several key enzymes that indicated adaptation of MAP to the host. Among these, FadE5, FadE25 and AdhB indicated that cholesterol is used as a carbon source in the bovine intestinal mucosa; the respiratory enzymes AtpA, NuoG and SdhA suggested increased respiration during infection. Furthermore higher levels of the pentose phosphate pathway enzymes Gnd2, Zwf and Tal as well as of KatG, SodA and GroEL indicated a vigorous stress response of MAP in vivo. In conclusion, our results provide novel insights into the metabolic adaptation of a pathogenic mycobacterium in its natural host. PMID:23223439

  16. Evidence for genes associated with the ability of Mycobacterium avium subsp. hominissuis to escape apoptotic macrophages

    PubMed Central

    Bermudez, Luiz E.; Danelishvili, Lia; Babrack, Lmar; Pham, Tuan

    2015-01-01

    Mycobacterium avium subsp. hominissuis (MAH) is an environmental bacteria that infects immunocompromised humans. MAH cases are increasing in incidence, making it crucial to gain knowledge of the pathogenic mechanisms associated with the bacterium. MAH infects macrophages and after several days the infection triggers the phagocyte apoptosis. Many of the intracellular MAH escape the cell undergoing apoptosis leading to infection of neighboring macrophages. We screened a transposon bank of MAH mutants in U937 mononuclear phagocytes for the inability to escape macrophages undergoing apoptosis. Mutations in genes; MAV_2235, MAV_2120, MAV_2410, and MAV_4563 resulted in the inability of the bacteria to exit macrophages upon apoptosis. Complementation of the mutations corrected the phenotype either completely or partially. Testing for the ability of the mutants to survive in macrophages compared to the wild-type bacterium revealed that the mutant clones were not attenuated up to 4 days of infection. Testing in vivo, however, demonstrated that all the MAH clones were attenuated compared with the wild-type MAC 104 in tissues of mice. Although the mechanism associated with the bacterial inability to leave apoptotic macrophages is unknown, the identification of macrophage cytoplasm targets for the MAH proteins suggest that they interfere either with protein degradation machinery or post-translation mechanisms. The identification of tatC as a MAH protein involved in the ability of MAH to leave macrophages, suggests that secreted effector(s) are involved in the process. The study reveals a pathway of escape from macrophages, not shared with Mycobacterium tuberculosis. PMID:26380226

  17. Discovery of novel small molecule modulators of Clavibacter michiganensis subsp. michiganensis

    PubMed Central

    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 ?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 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. PMID:26539169

  18. Evaluation and comparison of molecular techniques for epidemiological typing of Salmonella enterica subsp. enterica serovar dublin.

    PubMed Central

    Liebisch, B; Schwarz, S

    1996-01-01

    A total of 28 unrelated isolates of the Salmonella enterica subsp. enterica serovar dublin (S. dublin) collected during a 6-year period, as well as four samples of the S. dublin live vaccine strain Bovisaloral and its prototype strain S. dublin 442/039, were investigated by different molecular typing methods for the following reasons: (i) to find the most discriminatory method for the epidemiological typing of isolates belonging to this Salmonella serovar and (ii) to evaluate these methods for their capacity to discriminate among the live vaccine strain Bovisaloral, its prototype strain S. dublin 442/039, and field isolates of the serovar dublin. Five different plasmid profiles were observed; a virulence plasmid of 76 kbp as identified by hybridization with an spvB-spvC gene probe was present in all isolates. The detection of 16S rRNA genes and that of IS200 elements proved to be unsuitable for the epidemiological typing of S. dublin; only one hybridization pattern could be observed with each of these methods. The results obtained from macrorestriction analysis strongly depended on the choice of restriction enzyme. While the enzyme NotI yielded the lowest discriminatory index among all enzymes tested, it was the only enzyme that allowed discrimination between the Bovisaloral vaccine strain and its prototype strain. In contrast to the enzymes XbaI and SpeI, which only differentiated among the S. dublin field isolates, XhoI as well as AvrII also produced restriction fragment patterns of the Bovisaloral strain and of its prototype strain that were not shared by any of the S. dublin field isolates. Macrorestriction analysis proved to be the most discriminatory method not only for the epidemiological typing of S. dublin field isolates but also for the identification of the S. dublin live vaccine strain Bovisaloral. PMID:8904430

  19. Copper tolerance in the macrolichens Cladonia furcata and Cladina arbuscula subsp. mitis is constitutive rather than inducible.

    PubMed

    Ba?kor, Martin; Péli, Evelin Ramóna; Vantová, Ivana

    2011-09-01

    In this study we assessed the degree of copper (Cu) tolerance in two common lichen species (Cladonia furcata and Cladina arbuscula subsp. mitis) that grow on both uncontaminated substrata and the surface of waste heaps from abandoned old Cu-mines. Regardless of their locality, populations of these lichens contain identical strains of photobionts (Asterochloris clade A in C. arbuscula subsp. mitis and clade D in C. furcata). Therefore, it was expected that if there were differences in Cu toxicity or tolerance between populations, that the photobiont could not be a key element of Cu tolerance in these two lichen species. In laboratory experiments samples of both lichen species (from contaminated and control sites) were incubated in Cu solutions (500 ?M) for 24 h. We attempted to determine whether Cu tolerance in these lichens was constitutive, or inducible form. Based on measurements of Cu accumulation, chlorophyll a integrity, chlorophyll a fluorescence, photosynthesis, respiration, measurements of the content of thiobarbituric acid reactive substances (TBARS), the content of soluble proteins, reactive oxygen species (ROS) and the amount of extracellular secondary metabolites of both lichens we found that there were no significant differences in the response of all selected populations of both lichen species to short-term exposure to these high levels of Cu. As a result, we conclude that Cu tolerance in these two lichen species is the constitutive rather than the inducible. PMID:21676428

  20. Short- and long-term dynamics in the intestinal microbiota following ingestion of Bifidobacterium animalis subsp. lactis GCL2505

    PubMed Central

    TANAKA, Yoshiyuki; TAKAMI, Kazuyo; NISHIJIMA, Tomohiko; AOKI, Ryo; MAWATARI, Takashi; IKEDA, Takayuki

    2015-01-01

    Bifidobacterium animalis subsp. lactis GCL2505 (B. lactis GCL2505) is able to survive passage through the intestines and proliferate. The daily dynamics of the intestinal bifidobacteria following ingestion of probiotics are not yet clear. Moreover, the effects of long-term ingestion of probiotics on the intestinal microbiota have not been well studied. Two experiments were performed in the present study. In Experiment 1, 53 healthy female volunteers received B. lactis GCL2505; B. bifidum GCL2080, which can survive but not proliferate in the intestine; or yogurt fermented with Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus for 2 weeks, and the daily dynamics of intestinal bifidobacteria were investigated. The number of fecal bifidobacteria significantly increased on day 1, and this was maintained until day 14 in the B. lactis GCL2505 ingestion group. However, no significant change in the number of fecal bifidobacteria was observed in the other groups throughout the ingestion period. In Experiment 2, 38 constipated volunteers received either B. lactis GCL2505 or a placebo for 8 weeks. Both the number of fecal bifidobacteria and the frequency of defecation significantly increased throughout the ingestion period in the B. lactis GCL2505 ingestion group. These results suggested that the proliferation of ingested bifidobacteria within the intestine contributed to a rapid increase in the amount of intestinal bifidobacteria and subsequent maintenance of these levels. Moreover, B. lactis GCL2505 improved the intestinal microbiota more effectively than non-proliferating bifidobacteria and lactic acid bacteria. PMID:26594607

  1. Short- and long-term dynamics in the intestinal microbiota following ingestion of Bifidobacterium animalis subsp. lactis GCL2505.

    PubMed

    Tanaka, Yoshiyuki; Takami, Kazuyo; Nishijima, Tomohiko; Aoki, Ryo; Mawatari, Takashi; Ikeda, Takayuki

    2015-01-01

    Bifidobacterium animalis subsp. lactis GCL2505 (B. lactis GCL2505) is able to survive passage through the intestines and proliferate. The daily dynamics of the intestinal bifidobacteria following ingestion of probiotics are not yet clear. Moreover, the effects of long-term ingestion of probiotics on the intestinal microbiota have not been well studied. Two experiments were performed in the present study. In Experiment 1, 53 healthy female volunteers received B. lactis GCL2505; B. bifidum GCL2080, which can survive but not proliferate in the intestine; or yogurt fermented with Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus for 2 weeks, and the daily dynamics of intestinal bifidobacteria were investigated. The number of fecal bifidobacteria significantly increased on day 1, and this was maintained until day 14 in the B. lactis GCL2505 ingestion group. However, no significant change in the number of fecal bifidobacteria was observed in the other groups throughout the ingestion period. In Experiment 2, 38 constipated volunteers received either B. lactis GCL2505 or a placebo for 8 weeks. Both the number of fecal bifidobacteria and the frequency of defecation significantly increased throughout the ingestion period in the B. lactis GCL2505 ingestion group. These results suggested that the proliferation of ingested bifidobacteria within the intestine contributed to a rapid increase in the amount of intestinal bifidobacteria and subsequent maintenance of these levels. Moreover, B. lactis GCL2505 improved the intestinal microbiota more effectively than non-proliferating bifidobacteria and lactic acid bacteria. PMID:26594607

  2. Increased viability but decreased culturability of Mycobacterium avium subsp. paratuberculosis in macrophages from inflammatory bowel disease patients under Infliximab treatment.

    PubMed

    Nazareth, Nair; Magro, Fernando; Appelberg, Rui; Silva, Jani; Gracio, Daniela; Coelho, Rosa; Cabral, José Miguel; Abreu, Candida; Macedo, Guilherme; Bull, Tim J; Sarmento, Amélia

    2015-12-01

    Mycobacterium avium subsp. paratuberculosis (MAP) has long been implicated as a triggering agent in Crohn's disease (CD). In this study, we investigated the growth/persistence of both M. avium subsp. hominissuis (MAH) and MAP, in macrophages from healthy controls (HC), CD and ulcerative colitis patients. For viability assessment, both CFU counts and a pre16SrRNA RNA/DNA ratio assay (for MAP) were used. Phagolysosome fusion was evaluated by immunofluorescence, through analysis of LAMP-1 colocalization with MAP. IBD macrophages were more permissive to MAP survival than HC macrophages (a finding not evident with MAH), but did not support MAP active growth. The lower MAP CFU counts in macrophage cultures associated with Infliximab treatment were not due to increased killing, but possibly to elevation in the proportion of intracellular dormant non-culturable MAP forms, as MAP showed higher viability in those macrophages. Increased MAP viability was not related to lack of phagolysosome maturation. The predominant induction of MAP dormant forms by Infliximab treatment may explain the lack of MAP reactivation during anti-TNF therapy of CD but does not exclude the possibility of MAP recrudescence after termination of therapy. PMID:25702170

  3. Utilization of a thermosensitive episome bearing transposon TN10 to isolate Hfr donor strains of Erwinia carotovora subsp. chrysanthemi.

    PubMed Central

    Kotoujansky, A; Lemattre, M; Boistard, P

    1982-01-01

    A thermosensitive episome bearing the transposon Tn10, F(Ts)::Tn10 Lac+, has been successfully transferred from Escherichia coli to several wild strains of the enterobacteria Erwinia carotovora subsp. chrysanthemi, which are pathogenic on Saintpaulia ionantha. In one of these strains, all of the characters controlled by this episome (Lac+, Tetr, Tra+) were expressed, and its replication was stopped at 40 degrees C and above. At 30 degrees C, the episome was easily transferred between strains derived from E. carotovora subsp. chrysanthemi 3937j and to E coli. Hfr donor strains were obtained from a F' strain of 3937j by selecting clones which grew at 40 degrees C on plates containing tetracycline. One of these strains, Hfrq, was examined in more detail: the characters Lac+ and Tetr were stabilized and did not segregate higher than its parental F' strain. The mating was most efficient at 37 degrees C on a membrane. Hfrq transferred its chromosome to recipient strains at high frequency and in a polarized fashion, as evidenced by the gradient of transfer frequencies, the kinetics of marker entry (in interrupted mating experiments), and the analysis of linkage between different markers. The chromosome of Hfrq was most probably transferred in the following sequence: origin...met...xyl...arg...ile...leu...thr...cys...pan...ura...gal...trp...his. ..pur... Moreover, this genetic transfer system proved to be efficient in strain construction. PMID:6277860

  4. Immunogenicity and reactivity of novel Mycobacterium avium subsp. paratuberculosis PPE MAP1152 and conserved MAP1156 proteins with sera from experimentally and naturally infected animals

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s Disease (JD) in ruminants. Development of genetic tools and completion of the MAP genome sequencing project expanded opportunities for antigen discovery. In this study, we determined the seroreactivity of two proteins encoded for at th...

  5. The role of IL-10 in the regulation of the adaptive immune responses in cattle infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular bacterial pathogen that causes Johne’s disease in cattle and other animals. Infection follows ingestion of the bacteria primarily through the fecal oral route and results in the colonization of the intestine and a granulomatous en...

  6. Evaluation of a Granulovirus (PoGV) and Bacillus Thuringiensis subsp. kurstaki for Control of the Potato Tuberworm in Stored Tubers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Currently no insecticides are registered for postharvest use on potato tubers in North America. We evaluated liquid suspensions and dry formulations of a granulovirus (PoGV) and bacterium Bacillus thuringiensis subsp. kurstaki Berliner (Btk) for control of the potato tuber moth (PTM), Phthorimaea op...

  7. Mycobacterium avium subsp. paratuberculosis PPE Protein MAP1152 and Conserved Protein MAP1156 are Antigenic in Experimentally and Naturally Infected Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s Disease (JD) in ruminants resulting in significant production losses. An insertion mutation upstream from the MAP1152-MAP1156 region causes a change in colony morphotype and results in an attenuated phenotype in bovine monocyte derive...

  8. Modulation of Cytokine Expression and Lymphocyte Subsets during the Periparturient Period in Dairy Cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to evaluate cytokine gene expression and populations of lymphocyte subsets in periparturient dairy cows naturally infected with Mycobacterium avium subsp. paratuberculosis (MAP). Blood was collected from noninfected, subclinical, and clinical MAP-infected dairy cows ...

  9. Proteomic Comparison of the Cattle and Sheep Strains of Mycobacterium avium subsp. paratuberculosis in Different Growth Conditions Using iTRAQ Analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Paratuberculosis (Johne's disease) caused by Mycobacterium avium subsp. paratuberculosis (MAP) is a serious economic threat worldwide affecting cattle, sheep and goats. The infection can persist for several years in a subclinical phase that is difficult to identify because of the absence of clinical...

  10. Draft Genome Sequence for ICMP 5702, the Type Strain of Pectobacterium carotovorum subsp. carotovorum That Causes Soft Rot Disease on Potato

    PubMed Central

    Lu, Ashley; Armstrong, Karen F.

    2015-01-01

    Pectobacterium species are economically important bacteria that cause soft rotting of potato tubers in the field and in storage. Here, we report the draft genome sequence of the type strain for P. carotovorum subsp. carotovorum, ICMP 5702 (ATCC 15713). The genome sequence of ICMP 5702 will provide an important reference for future phylogenomic and taxonomic studies of the phytopathogenic Enterobacteriaceae. PMID:26251498

  11. Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium subsp. paratuberculosis, milk production and longevity in Holstein dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In summer 2007, a randomized controlled clinical trial was initiated on 6 large Midwest commercial dairy farms to investigate the effect of feeding heat-treated (HT) colostrum on transmission of Mycobacterium avium subsp. paratuberculosis and on future milk production and longevity within the herd. ...

  12. Optimization of methods for the detection of Mycobacterium avium subsp. paratuberculosis in milk and colostrum of naturally infected dairy cows with Johne's Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s Disease (JD), a chronic enteritis that occurs in dairy cattle and other ruminants. A 2007 NAHMS Dairy Study demonstrated that over 68% of dairy herds are infected with JD so the risk of exposure within a herd is high...

  13. Optimization of Methods for Obtaining, Extracting and Detecting Mycobacterium avium subsp. paratuberculosis in Environmental Samples using Quantitative, Real-Time PCR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Detection of Johne’s disease, an enteric infection of cattle caused by Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), has been impeded by the lack of rapid, reliable detection methods. The goal of this study was to optimize methodologies for obtaining, extracting and evaluating t...

  14. Evaluation of the association between fecal excretion of Mycobacterium avium subsp paratuberculosis and detection in colostrum and on teat skin surfaces of dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objective—To evaluate the association between fecal excretion of Mycobacterium avium subsp paratuberculosis (MAP) by dairy cows in the periparturient period and detection of MAP DNA in colostrum specimens and on teat skin surfaces. Design—Cross-sectional study. Animals—112 Holstein cows. Procedures—...

  15. Depletion of CD4 T Lymphocytes at the time of infection with M. avium subsp. paratuberculosis does not accelerate disease progression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A calf model was used to determine if the depletion of CD4 T cells prior to inoculation of Mycobacterium avium subsp. paratuberculosis (Map) would delay development of an immune response to Map and accelerate disease progression. Ileal cannulas were surgically implanted in 5 bull calves at two month...

  16. Use of carnauba based carrier for copper sprays reduces infection by Xanthomonas citri subsp. citri and Diaporthe citri in Florida commercial grapefruit groves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Asiatic citrus canker caused by Xanthomonas citri subsp. citri (Xcc), a bacterial disease of citrus, was first documented on Florida citrus in the early 1900’s. At that time the disease was managed, only to return in the 1980’s and 90’s and to finally remain uncontrolled in 2004. Xcc is most active ...

  17. Association of single nucleotide polymorphisms in the ANKRA2 and CD180 genes with bovine respiratory disease and presence of Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective was to determine if single nucleotide polymorphisms (SNPs) in the ANKRA2 and CD180 genes were associated with incidence of bovine respiratory disease (BRD) and presence of Mycobacterium avium subsp. paratuberculosis (MAP) in cattle. Two independent populations were used. The first po...

  18. Genome Sequences of Three Salmonella enterica subsp. enterica Serovar Infantis Strains from Healthy Broiler Chicks in Hungary and in the United Kingdom.

    PubMed

    Olasz, Ferenc; Nagy, Tibor; Szabó, Móni; Kiss, János; Szmolka, Ama; Barta, Endre; van Tonder, Andries; Thomson, Nicholas; Barrow, Paul; Nagy, Béla

    2015-01-01

    The genome sequences of three strains of Salmonella enterica subsp. enterica serovar Infantis isolated from broiler chickens in 1994 and 2004 in Hungary and in the 1980s in the United Kingdom are reported here. A sequence comparison should improve our understanding of the evolution of the genome and spread of S. Infantis in poultry. PMID:25676749

  19. Simultaneous Detection of Acidovorax Avenae Subsp. Citrulli and Didymella Bryoniae in Cucurbit Seedlots Using Magnetic Capture Hybridization and Real-Time Polymerase Chain Reaction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To improve the detection of plant pathogens in seeds magnetic capture hybridization and multiplex real-time PCR (MCH real-time PCR) was evaluated. Single stranded DNA hybridization capture probes targeting Acidovorax avenae subsp. citrulli and Didymella bryoniae DNA were covalently attached to magn...

  20. Clinical disease and stage of lactation influences shedding of Mycobacterium avium subsp. paratuberculosis into milk and colostrum of naturally infected dairy cows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease (JD). One mode of transmission of MAP is through ingestion of contaminated milk and colostrum by susceptible calves. The objective of this study was to determine if the amount of MAP shed into the milk and co...

  1. Complete Genome and Plasmid Sequences of Three Canadian Strains of Salmonella enterica subsp. enterica Serovar Enteritidis Belonging to Phage Types 8, 13, and 13a.

    PubMed

    Rehman, Muhammad Attiq; Labbé, Geneviève; Ziebell, Kim; Johnson, Roger P; Nash, John H E

    2015-01-01

    Salmonella enterica subsp. enterica serovar Enteritidis is a prominent cause of human salmonellosis frequently linked to poultry products. In Canada, S. Enteritidis phage types 8, 13, and 13a predominate among both clinical and poultry isolates. Here, we report the complete genome and plasmid sequences of poultry isolates of these three phage types. PMID:26404595

  2. Isolation of High-Affinity Single-Chain Antibodies against Mycobacterium avium subsp. paratuberculosis Surface Proteins from Sheep with Johne's Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Johne’s disease, due to infection with Mycobacterium avium subsp. paratuberculosis, causes significant economic losses to the livestock farming industry. Improved investigative and diagnostic tools - necessary to understand disease processes and to identify sub-clinical infection - are much sought a...

  3. Draft Genome Sequence of the Moderately Heat-Tolerant Lactococcus lactis subsp. lactis bv. diacetylactis Strain GL2 from Algerian Dromedary Milk.

    PubMed

    Gabed, Noujoud; Yang, Manli; Bey Baba Hamed, Mohamed; Drici, Habiba; Gross, Roy; Dandekar, Thomas; Liang, Chunguang

    2015-01-01

    Lactococcus lactis subsp. lactis bv. diacetylactis GL2 is a moderately thermotolerant lactic acid bacterium isolated from dromedary raw milk. Here, we present the draft genome sequence of this potential new dairy starter strain, which combines thermotolerance and the capacity to metabolize lactose, casein, and citrate. PMID:26586883

  4. Draft Genome Sequence of the Moderately Heat-Tolerant Lactococcus lactis subsp. lactis bv. diacetylactis Strain GL2 from Algerian Dromedary Milk

    PubMed Central

    Gabed, Noujoud; Yang, Manli; Bey Baba Hamed, Mohamed; Drici, Habiba; Gross, Roy; Dandekar, Thomas

    2015-01-01

    Lactococcus lactis subsp. lactis bv. diacetylactis GL2 is a moderately thermotolerant lactic acid bacterium isolated from dromedary raw milk. Here, we present the draft genome sequence of this potential new dairy starter strain, which combines thermotolerance and the capacity to metabolize lactose, casein, and citrate. PMID:26586883

  5. Modulation of Cytokine Gene Expression and Secretion during the Periparturient Period in Dairy cows Naturally Infected with Mycobacterium avium subsp. paratuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Twenty multiparous and two primiparous Holstein cows were grouped according to infection status with Mycobacterium avium subsp. paratuberculosis (MAP), the causative microorganism for Johne’s disease. The effect of parturition and infection on the progression of Johne’s disease was monitored by det...

  6. Genotypic Characterization of Streptococcus infantarius subsp. coli Isolates from Sea Otters with Infective Endocarditis and/or Septicemia and from Environmental Mussel Samples

    PubMed Central

    Counihan-Edgar, Katrina L.; Gill, Verena A.; Doroff, Angela M.; Burek, Kathleen A.; Miller, Woutrina A.; Shewmaker, Patricia L.; Jang, Spencer; Goertz, Caroline E. C.; Tuomi, Pamela A.; Miller, Melissa A.; Jessup, David A.

    2012-01-01

    Pulsed-field gel electrophoresis (PFGE) was used to type 128 Streptococcus infantarius subsp. coli isolates from sea otters and mussels. Six SmaI PFGE groups were detected, with one predominant group representing 57% of the isolates collected over a wide geographic region. Several sea otter and mussel isolates were highly related, suggesting that an environmental infection source is possible. PMID:23052307

  7. Bacillus amyloliquefaciens subsp. plantarum GR53, a potent biocontrol agent resists Rhizoctonia disease on Chinese cabbage through hormonal and antioxidants regulation.

    PubMed

    Kang, Sang-Mo; Radhakrishnan, Ramalingam; Lee, In-Jung

    2015-10-01

    The fungus Rhizoctonia solani is one of the causal agents of numerous diseases that affect crop growth and yield. The aim of this present investigation was to identify a biocontrol agent that acts against R. solani and to determine the agent's protective effect through phytohormones and antioxidant regulation in experimentally infected Chinese cabbage plants. Four rhizospheric soil bacterial isolates GR53, GR169, GR786, and GR320 were tested for their antagonistic activity against R. solani. Among these isolates, GR53 significantly suppressed fungal growth. GR53 was identified as Bacillus amyloliquefaciens subsp. plantarum by phylogenetic analysis of the 16S rDNA sequence. The biocontrol activity of B. amyloliquefaciens subsp. plantarum GR53 was tested in Chinese cabbage plants under controlled conditions. Results showed that R. solani inhibited plant growth (length, width, fresh and dry weight of leaves) by reducing chlorophyll and total phenolic content, as well as by increasing the levels of salicylic acid, jasmonic acid, abscisic acid, and DPPH scavenging activity. By regulating the levels of these compounds, the co-inoculation of B. amyloliquefaciens subsp. plantarum GR53 heightened induced systemic resistance in infected Chinese cabbage, effectively mitigating R. solani-induced damaging effects and improving plant growth. The results obtained from this study suggest that B. amyloliquefaciens subsp. plantarum GR53 is an effective biocontrol agent to prevent the damage caused by R. solani in Chinese cabbage plants. PMID:26160009

  8. Analysis of the Immune Response to a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis in Experimentally and Naturally Infected Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 35 kDa major membrane protein (MMP) of Mycobacterium avium subsp. paratuberculosis (Map) is implicated in the pathogenesis of paratuberculosis (Ptb) in cattle. Understanding the immune response to MMP could reveal how Map evades immune elimination and provide information needed for developing a ...

  9. The Rhizobacterium Bacillus amyloliquefaciens subsp. plantarum NAU-B3 Contains a Large Inversion within the Central Portion of the Genome

    PubMed Central

    Wu, Huijun; Qiao, Junqing; Blom, Jochen; Rueckert, Christian; Reva, Oleg

    2013-01-01

    The genome of rhizobacterium Bacillus amyloliquefaciens subsp. plantarum strain NAU-B3 is 4,196,170 bp in size and harbors 4,001 genes. Nine giant gene clusters are dedicated to the nonribosomal synthesis of antimicrobial lipopeptides and polyketides. Remarkably, NAU_B3 contains a large inversion within the central portion of the genome. PMID:24233590

  10. The complete genome of Bacillus amyloliquefaciens subsp. plantarum CAU B946 contains a gene cluster for nonribosomal synthesis of iturin A.

    PubMed

    Blom, Jochen; Rueckert, Christian; Niu, Ben; Wang, Qi; Borriss, Rainer

    2012-04-01

    The genome of the rhizobacterium Bacillus amyloliquefaciens subsp. plantarum CAU B946 was 4.02 Mb in size and harbored 3,823 genes (coding sequences [CDS]). Nine giant gene clusters were dedicated to nonribosomal synthesis of antimicrobial compounds. Remarkably, strain CAU B946 possessed a gene cluster involved in synthesis of iturin A. PMID:22408246

  11. Diversity and Homogeneity among Small Plasmids of Aeromonas salmonicida subsp. salmonicida Linked with Geographical Origin

    PubMed Central

    Attéré, Sabrina A.; Vincent, Antony T.; Trudel, Mélanie V.; Chanut, Romain; Charette, Steve J.

    2015-01-01

    Furunculosis, which is caused by Aeromonas salmonicida subsp. salmonicida, is a major salmonid disease in fish farms worldwide. Several plasmids found in this bacterium confer phenotypes such drug resistance and virulence. Small plasmids (pAsa1, pAsa2, pAsa3, and pAsal1) related to ColE1- and ColE2-type replicons are usually present in its normal plasmidome. In the present study, with the objective to investigate if these plasmids display particularities related to the origin of the isolates bearing them, a total of 153 isolates, including 78 new and 75 previously described, were analyzed for the presence of small plasmids by PCR and DNA restriction fragment profiling. A geographical dichotomy between Canadian and European isolates for their propensity to do not have pAsa3 or pAsal1 was found. In addition, the genotyping analysis led to the identification of two European isolates harboring an unusual pAsal1. An investigation by next-generation sequencing (NGS) of these two isolates shed light on two pAsal1 variants (pAsal1C and pAsal1D). As with pAsal1B, another pAsal1 variant previously described, these two new variants bore a second insertion sequence (ISAS5) in addition to the usual ISAS11. The characterization of these variants suggested that they could predominate over the wild-type pAsal1 in stressful conditions such as growth at temperatures of 25°C and above. To obtain a comprehensive portrait of the mutational pressure on small plasmids, 26 isolates whose DNA had been sequenced by NGS were investigated. pAsa3 and pAsal1 were more prone to mutations than pAsa1 and pAsa2, especially in the mobA gene, which encodes a relaxase and a primase. Lastly, the average copy number of each plasmid per cell was assessed using raw sequencing data. A clear trend with respect to the relative proportion per cell of each plasmid was identified. Our large-scale study revealed a geographical dichotomy in small plasmid repertoire in addition to a clear trend for pAsa3 and pAsal1 to be more frequently altered. Moreover, we present the discovery of two new variants of pAsal1: pAsal1C and pAsal1D. PMID:26635745

  12. Fate of Mycobacterium avium subsp. paratuberculosis after application of contaminated dairy cattle manure to agricultural soils.

    PubMed

    Salgado, M; Collins, M T; Salazar, F; Kruze, J; Bölske, G; Söderlund, R; Juste, R; Sevilla, I A; Biet, F; Troncoso, F; Alfaro, M

    2011-03-01

    Details regarding the fate of Mycobacterium avium subsp. paratuberculosis (basonym, Mycobacterium paratuberculosis) after manure application on grassland are unknown. To evaluate this, intact soil columns were collected in plastic pipes (lysimeters) and placed under controlled conditions to test the effect of a loamy or sandy soil composition and the amount of rainfall on the fate of M. paratuberculosis applied to the soil surface with manure slurry. The experiment was organized as a randomized design with two factors and three replicates. M. paratuberculosis-contaminated manure was spread on the top of the 90-cm soil columns. After weekly simulated rainfall applications, water drainage samples (leachates) were collected from the base of each lysimeter and cultured for M. paratuberculosis using Bactec MGIT ParaTB medium and supplements. Grass was harvested, quantified, and tested from each lysimeter soil surface. The identity of all probable M. paratuberculosis isolates was confirmed by PCR for IS900 and F57 genetic elements. There was a lag time of 2 months after each treatment before M. paratuberculosis was found in leachates. The greatest proportions of M. paratuberculosis-positive leachates were from sandy-soil lysimeters in the manure-treated group receiving the equivalent of 1,000 mm annual rainfall. Under the higher rainfall regimen (2,000 mm/year), M. paratuberculosis was detected more often from lysimeters with loamy soil than sandy soil. Among all lysimeters, M. paratuberculosis was detected more often in grass clippings than in lysimeter leachates. At the end of the trial, lysimeters were disassembled and soil cultured at different depths, and we found that M. paratuberculosis was recovered only from the uppermost levels of the soil columns in the treated group. Factors associated with M. paratuberculosis presence in leachates were soil type and soil pH (P < 0.05). For M. paratuberculosis presence in grass clippings, only manure application showed a significant association (P < 0.05). From these findings we conclude that this pathogen tends to move slowly through soils (faster through sandy soil) and tends to remain on grass and in the upper layers of pasture soil, representing a clear infection hazard for grazing livestock and a potential for the contamination of runoff after heavy rains. PMID:21239557

  13. Lactase persistence, NOD2 status and Mycobacterium avium subsp. paratuberculosis infection associations to Inflammatory Bowel Disease

    PubMed Central

    2012-01-01

    Background Inflammatory Bowel Disease (IBD), which includes both Crohn’s disease (CD) and ulcerative colitis (UC), is caused by a complex interplay involving genetic predisposition, environmental factors and an infectious agent. Mycobacterium avium subsp. paratuberculosis (MAP) is a promising pathogen candidate since it produces a chronic intestinal inflammatory disease in ruminants that resembles CD in humans. MAP is a ubiquitous microorganism, although its presence in the food chain, especially in milk from infected animals, is what made us think that there could be an association between lactase persistence (LP) and IBD. The LCT mutation has brought adaptation to dairy farming which in turn would have increased exposure of the population to infection by MAP. NOD2 gene mutations are highly associated to CD. Methods In our study, CD and UC patients and controls from the North of Spain were genotyped for the lactase gene (LCT) and for three NOD-2 variants, R702W, G908R and Cins1007fs. MAP PCR was carried out in order to assess MAP infection status and these results were correlated with LCT and NOD2 genotypes. Results As for LP, no association was found with IBD, although UC patients were less likely to present the T/T?13910 variant compared to controls, showing a higher C-allele frequency and a tendency to lactase non-persistence (LNP). NOD2 mutations were associated to CD being the per-allele risk higher for the Cins1007fs variant. MAP infection was more extended among the healthy controls (45.2%) compared to CD patients (21.38%) and UC patients (19.04%) and this was attributed to therapy. The Asturian CD cohort presented higher levels of MAP prevalence (38.6%) compared to the Basque CD cohort (15.5%), differences also attributed to therapy. No interaction was found between MAP infection and LCT or NOD2 status. Conclusions We conclude that LP is not significantly associated with IBD, but that MAP infection and NOD2 do show not mutually interacting associations with IBD. PMID:22742424

  14. Development and Evaluation of a Novel Multicopy-Element-Targeting Triplex PCR for Detection of Mycobacterium avium subsp. paratuberculosis in Feces

    PubMed Central

    Garrido, Joseba M.; Molina, Elena; Geijo, María V.; Elguezabal, Natalia; Vázquez, Patricia; Juste, Ramón A.

    2014-01-01

    The enteropathy called paratuberculosis (PTB), which mainly affects ruminants and has a worldwide distribution, is caused by Mycobacterium avium subsp. paratuberculosis. This disease significantly reduces the cost-effectiveness of ruminant farms, and therefore, reliable and rapid detection methods are needed to control the spread of the bacterium in livestock and in the environment. The aim of this study was to identify a specific and sensitive combination of DNA extraction and amplification to detect M. avium subsp. paratuberculosis in feces. Negative bovine fecal samples were inoculated with increasing concentrations of two different bacterial strains (field and reference) to compare the performance of four extraction and five amplification protocols. The best results were obtained using the JohnePrep and MagMax extraction kits combined with an in-house triplex real-time PCR designed to detect IS900, ISMap02 (an insertion sequence of M. avium subsp. paratuberculosis present in 6 copies per genome), and an internal amplification control DNA simultaneously. These combinations detected 10 M. avium subsp. paratuberculosis cells/g of spiked feces. The triplex PCR detected 1 fg of genomic DNA extracted from the reference strain K10. The performance of the robotized version of the MagMax extraction kit combined with the IS900 and ISMap02 PCR was further evaluated using 615 archival fecal samples from the first sampling of nine Friesian cattle herds included in a PTB control program and followed up for at least 4 years. The analysis of the results obtained in this survey demonstrated that the diagnostic method was highly specific and sensitive for the detection of M. avium subsp. paratuberculosis in fecal samples from cattle and a very valuable tool to be used in PTB control programs. PMID:24727272

  15. Identification of Three Novel Superantigen-Encoding Genes in Streptococcus equi subsp. zooepidemicus, szeF, szeN, and szeP? †

    PubMed Central

    Paillot, Romain; Darby, Alistair C.; Robinson, Carl; Wright, Nicola L.; Steward, Karen F.; Anderson, Emma; Webb, Katy; Holden, Matthew T. G.; Efstratiou, Androulla; Broughton, Karen; Jolley, Keith A.; Priestnall, Simon L.; Marotti Campi, Maria C.; Hughes, Margaret A.; Radford, Alan; Erles, Kerstin; Waller, Andrew S.

    2010-01-01

    The acquisition of superantigen-encoding genes by Streptococcus pyogenes has been associated with increased morbidity and mortality in humans, and the gain of four superantigens by Streptococcus equi is linked to the evolution of this host-restricted pathogen from an ancestral strain of the opportunistic pathogen Streptococcus equi subsp. zooepidemicus. A recent study determined that the culture supernatants of several S. equi subsp. zooepidemicus strains possessed mitogenic activity but lacked known superantigen-encoding genes. Here, we report the identification and activities of three novel superantigen-encoding genes. The products of szeF, szeN, and szeP share 59%, 49%, and 34% amino acid sequence identity with SPEH, SPEM, and SPEL, respectively. Recombinant SzeF, SzeN, and SzeP stimulated the proliferation of equine peripheral blood mononuclear cells, and tumor necrosis factor alpha (TNF-?) and gamma interferon (IFN-?) production, in vitro. Although none of these superantigen genes were encoded within functional prophage elements, szeN and szeP were located next to a prophage remnant, suggesting that they were acquired by horizontal transfer. Eighty-one of 165 diverse S. equi subsp. zooepidemicus strains screened, including 7 out of 15 isolates from cases of disease in humans, contained at least one of these new superantigen-encoding genes. The presence of szeN or szeP, but not szeF, was significantly associated with mitogenic activity in the S. equi subsp. zooepidemicus population (P < 0.000001, P < 0.000001, and P = 0.104, respectively). We conclude that horizontal transfer of these novel superantigens from and within the diverse S. equi subsp. zooepidemicus population is likely to have implications for veterinary and human disease. PMID:20713629

  16. Immunological evaluation and cellular location analysis of the TprI antigen of Treponema pallidum subsp. pallidum.

    PubMed

    Giacani, Lorenzo; Sambri, Vittorio; Marangoni, Antonella; Cavrini, Francesca; Storni, Elisa; Donati, Manuela; Corona, Silvia; Lanzarini, Paolo; Cevenini, Roberto

    2005-06-01

    The TprI antigen of Treponema pallidum subsp. pallidum is a putative virulence factor predicted to be located in the outer membrane of the syphilis spirochete. In this study, we analyzed the immune response against TprI and its subunits in sera collected both from rabbits experimentally infected with the Nichols strain and from patients with syphilis, showing a different pattern of reactivity toward the antigen in these two groups of samples. The protective ability of recombinant TprI and its hypothetical outer membrane location were also investigated. Although no rabbit was protected after challenge, immunoelectron microscopy results, to be further investigated, were compatible with the outer membrane location of the antigen. PMID:15908421

  17. Topical Allium ampeloprasum subsp Iranicum (Leek) extract cream in patients with symptomatic hemorrhoids: a pilot randomized and controlled clinical trial.

    PubMed

    Mosavat, Seyed Hamdollah; Ghahramani, Leila; Sobhani, Zahra; Haghighi, Ehsan Rahmanian; Heydari, Mojtaba

    2015-04-01

    Allium ampeloprasum subsp iranicum (Leek) has been traditionally used in antihemorrhoidal topical herbal formulations. This study aimed to evaluate its safety and efficacy in a pilot randomized controlled clinical trial. Twenty patients with symptomatic hemorrhoids were randomly allocated to receive the topical leek extract cream or standard antihemorrhoid cream for 3 weeks. The patients were evaluated before and after the intervention in terms of pain, defecation discomfort, bleeding severity, anal itching severity, and reported adverse events. A significant decrease was observed in the grade of bleeding severity and defecation discomfort in both the leek and antihemorrhoid cream groups after the intervention, while no significant change was observed in pain scores. There was no significant difference between the leek and antihemorrhoid cream groups with regard to mean changes in outcome measures. This pilot study showed that the topical use of leek cream can be as effective as a standard antihemorrhoid cream. PMID:25608984

  18. Biological Activities of Various Extracts and Chemical Composition of Trigonella monantha C. A. Mey. subsp. monantha Grown in Iran

    PubMed Central

    Esmaeili, Akbar; Rashidi, Bahareh; Rezazadeh, Shamsali

    2012-01-01

    The objective of this research was to study the biological activities of various extracts and the chemical composition of the essential oil Trigonella monantha C. A. Mey. subsp. monantha by gas chromatography (GC) and gas chromatography-mass spectroscopy (GC-MS), from the compounds derived from the aerial parts. The overall results of T. monantha tests allow us to conclude various extracts [hexane extract (HE), methanol extract (ME) and chloroform extract (CE)] of the test total phenolic, ABTS and DPPH while also testing ?-carotene largest property antioxidant. The property antioxidant shows the extracts of both mechanisms of electron transfer and the hydrogen transfer it has gone through. The antimicrobial activity of the extracts of both samples was determined against seven Gram-positive and Gram-negative bacteria. The strongest activity exhibited by the T. monantha of ME was determined to be 49.58 ?g/mL and was exhibited by ME. PMID:24250546

  19. Purification and thermostability of beta-galactosidase (lactase) from an autolytic strain of Streptococcus salivarius subsp. thermophilus.

    PubMed

    Chang, B S; Mahoney, R R

    1989-02-01

    beta-Galactosidase from an autolytic strain of Streptococcus salivarius subsp. thermophilus was purified 109-fold to near homogeneity. The yield of purified enzyme was 41% and the specific activity was 592 o-nitrophenyl beta-D-galactopyranoside U/mg at 37 degrees C. Two isozymes were present, but only one subunit was detected, having a mol. wt of 116,000. Enzyme stability was 37-83 times greater in milk than in buffer in the range 60-65 degrees C. At 60 degrees C the half-life in milk was 146 min. Denaturation in buffer was first-order, but in milk the overall reaction order with respect to enzyme concentration was approximately 0.5. The activation energy for denaturation was 453 kJ/mol in milk and 372 kJ/mol in buffer. In milk the activation energy for lactose hydrolysis was 35.1 kJ/mol. PMID:2495313

  20. Conjugative plasmid in Corynebacterium flaccumfaciens subsp. oortii that confers resistance to arsenite, arsenate, and antimony(III)

    SciTech Connect

    Hendrick, C.A.; Haskins, W.P.; Vidaver, A.K.

    1984-07-01

    Gene transfer systems for phytopathogenic corynebacteria have not been reported previously. In this paper a conjugative 46-megadalton plasmid (pDG101) found in Corynebacterium flaccumfaciens subsp. oorii CO101 is described that mediates resistance to arsenite, arsenate, and antimony(III). Transfer of the plasmid from CO101 to four other strains from the C. flaccumfaciens group occurred between cells immobilized on nitrocellulose filters or on agar surfaces. Transconjugant strains expressed the same levels of metal resistance as the donor strain and were able to act as donor strains in subsequent matings. The physical presence of the plasmid was detected by agarose gel electrophoresis. Arsenite-sensitive derivatives of the donor and transconjugant strains were obtained after heat treatment; these were cured of pDG101.

  1. An outbreak of granulomatous inflammation associated with Francisella noatunensis subsp. orientalis in farmed tilapia (Oreochromis niloticus × O. aureus) in China

    NASA Astrophysics Data System (ADS)

    Lin, Qiang; Li, Ningqiu; Fu, Xiaozhe; Hu, Qiandong; Chang, Ouqin; Liu, Lihui; Zhang, Defeng; Wang, Guangjun; San, Guibao; Wu, Shuqin

    2015-10-01

    In 2013, a novel disease was detected in tilapia (Oreochromis niloticus × O. aureus) in Guangzhou, South China. To identify the causative pathogen, we conducted histological examination, bacteria isolation, and purification, and sequenced the 16S rRNA gene of isolates. Infected fish had a large number of white granulomas in the kidney, liver, heart, and spleen. The head kidney and spleen were markedly swollen. A bacterium strain designated as gz201301 was recovered from the spleen of infected tilapia. The 16S rRNA sequence of gz201301 revealed that it was highly similar to F. noatunensis subsp. orientalis. This is the first report of a Francisella-like infection in farmed tilapia in China.

  2. Operational Evaluation Of Vectomax® WSP (Bacillus thuringiensis Subsp. israelensis+Bacillus sphaericus) Against Larval Culex pipiens in Septic Tanks (1).

    PubMed

    Cetin, Huseyin; Oz, Emre; Yanikoglu, Atila; Cilek, James E

    2015-06-01

    The residual effectiveness of VectoMax® WSP (a water-soluble pouch formulation containing a combination of Bacillus thuringiensis subsp. israelensis strain AM65-52 and B. sphaericus strain ABTS 1743) when applied to septic tanks against 3rd- and 4th-stage larvae of Culex pipiens L. was evaluated in this study. This formulation was evaluated at operational application rates of 1 pouch (10 g) and 2 pouches (20 g) per septic tank. Both application rates resulted in >96% control of larvae for 24 days. Operationally, VectoMax WSP has proven to be a useful tool for the nonchemical control of Culex species in septic tank environments. PMID:26181699

  3. Cistus creticus subsp. eriocephalus as a Model for Studying Plant Physiological and Metabolic Responses to Environmental Stress Factors.

    PubMed

    Paolessi, Paola; Nicoletti, Marcello; Catoni, Rosangela; Puglielli, Giacomo; Toniolo, Chiara; Gratani, Loretta

    2015-12-01

    Variations in physiology and metabolic products of Cistus creticus subsp. eriocephalus along an altitudinal gradient (350-750?m.a.s.l.) within the Monti Lucretili Regional Natural Park (central Italy) were studied. The results showed that the phenol production was in relationship with the net photosynthetic rates and the chlorophyll content. In particular, the increasing caffeic acid (CA) content with altitude suggested its role in providing an additional photo-protection mechanism, by its ability to consume photochemical reducing power and acting as an alternative C-atom sink under high light conditions. The metabolic production was tested by high performance thin layer chromatography (HPTLC) fingerprint analysis, highlighting the potential of this technique in biologic studies. PMID:26663839

  4. Comprehensive Insights in the Mycobacterium avium subsp. paratuberculosis Genome Using New WGS Data of Sheep Strain JIII-386 from Germany

    PubMed Central

    Möbius, Petra; Hölzer, Martin; Felder, Marius; Nordsiek, Gabriele; Groth, Marco; Köhler, Heike; Reichwald, Kathrin; Platzer, Matthias; Marz, Manja

    2015-01-01

    Mycobacterium avium (M. a.) subsp. paratuberculosis (MAP)—the etiologic agent of Johne’s disease—affects cattle, sheep, and other ruminants worldwide. To decipher phenotypic differences among sheep and cattle strains (belonging to MAP-S [Type-I/III], respectively, MAP-C [Type-II]), comparative genome analysis needs data from diverse isolates originating from different geographic regions of the world. This study presents the so far best assembled genome of a MAP-S-strain: Sheep isolate JIII-386 from Germany. One newly sequenced cattle isolate (JII-1961, Germany), four published MAP strains of MAP-C and MAP-S from the United States and Australia, and M. a. subsp. hominissuis (MAH) strain 104 were used for assembly improvement and comparisons. All genomes were annotated by BacProt and results compared with NCBI (National Center for Biotechnology Information) annotation. Corresponding protein-coding sequences (CDSs) were detected, but also CDSs that were exclusively determined by either NCBI or BacProt. A new Shine–Dalgarno sequence motif (5?-AGCTGG-3?) was extracted. Novel CDSs including PE-PGRS family protein genes and about 80 noncoding RNAs exhibiting high sequence conservation are presented. Previously found genetic differences between MAP-types are partially revised. Four of ten assumed MAP-S-specific large sequence polymorphism regions (LSPSs) are still present in MAP-C strains; new LSPSs were identified. Independently of the regional origin of the strains, the number of individual CDSs and single nucleotide variants confirms the strong similarity of MAP-C strains and shows higher diversity among MAP-S strains. This study gives ambiguous results regarding the hypothesis that MAP-S is the evolutionary intermediate between MAH and MAP-C, but it clearly shows a higher similarity of MAP to MAH than to Mycobacterium intracellulare. PMID:26384038

  5. Comparison of bacterial culture and polymerase chain reaction (PCR) for the detection of F. tularensis subsp. holarctica in wild animals.

    PubMed

    Sting, Reinhard; Runge, Martin; Eisenberg, Tobias; Braune, Silke; Müller, Wolfgang; Otto, Peter

    2013-01-01

    Detection of the zoonotic pathogen Francisella tularensis subsp. holarctica (EF tularensis) in wild animals with culture techniques as well as polymerase chain reaction were compared and discussed on the basis of the investigation of 60 animals. The samples originated from 55 European brown hares (Lepus europaeus), two red foxes (Vulpes vulpes) and one each from a wild rabbit (Oryctolagus cuniculus), a European beaver (Castor fiber), and a lemur (Lemur catta). When comparing the growth of 28 F. tularensis isolates on the cysteine blood agar and the modified Martin-Lewis-agar used in this study, cultivation was successful for 26 isolates on both media, but for two isolates only on the cysteine blood agar. Out of 43 carcasses 19 tested positive in bacteriological culture and PCR. Two culture positive samples of tonsils originating from foxes could not be confirmed by PCR, although PCR was positive in 22 samples that missed growth of F. tularensis. Comparative studies on cultural detection of E. tularensis were performed on samples of 16 hares from lung, spleen, liver and gut and in one case with a peritoneal swab. In at least one of these localizations cultivation of the pathogen was successful. Detection rate was reduced to 94% (15 of 16 hares) considering only the results of the cultures of the lungs and spleens. For a sensitive and rapid detection of F. tularensis subsp. holarctica, the PCR is a suitable method thereby avoiding hazardous multiplying of the pathogen. However, cultivation of F. tularensis is often a prerequisite for further studies on antibiotic resistance patterns of the pathogen, molecular epidemiological and pathological analyses of tularaemia. PMID:23901583

  6. In silico epitope analysis of unique and membrane associated proteins from Mycobacterium avium subsp. paratuberculosis for immunogenicity and vaccine evaluation.

    PubMed

    Carlos, Perla; Roupie, Virginie; Holbert, Sébastien; Ascencio, Felipe; Huygen, Kris; Gomez-Anduro, Gracia; Branger, Maxime; Reyes-Becerril, Martha; Angulo, Carlos

    2015-11-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis disease affecting ruminants worldwide. The aim of this study was to identify potential candidate antigens and epitopes by bio and immuno-informatic tools which could be later evaluated as vaccines and/or diagnosis. 110 protein sequences were selected from MAP K-10 genome database: 48 classified as putative enzymes involved in surface polysaccharide and lipopolysaccharide synthesis, as membrane associated and secreted proteins, 32 as conserved membrane proteins, and 30 as absent from other mycobacterial genomes. These 110 proteins were preliminary screened for Major Histocompatibility Complex (MHC) class II affinity and promiscuity using ProPred program. In addition, subcellular localization and host protein homology was analyzed. From these analyses, 23 MAP proteins were selected for a more accurate inmunoinformatic analysis (i.e. T cell and B cell epitopes analysis) and for homology with mycobacterial proteins. Finally, eleven MAP proteins were identified as potential candidates for further immunogenic evaluation: six proteins (MAP0228c, MAP1239c, MAP2232, MAP3080, MAP3131 and MAP3890) were identified as presenting potential T cell epitopes, while 5 selected proteins (MAP0232c, MAP1240c, MAP1738, MAP2239 and MAP3641c) harbored a large numbers of epitopes predicted to induce both cell- and antibody-mediated immune responses. Moreover, immunogenicity of selected epitopes from MAP1239c were evaluated in IFN-? release assay. In summary, eleven M. avium subsp. paratuberculosis proteins were identified by in silico analysis and need to be further evaluated for their immunodiagnostic and vaccine potential in field and mice model. PMID:26279134

  7. Chemical composition and anti-biofilm activity of Thymus sipyleus BOISS. subsp. sipyleus BOISS. var. davisianus RONNIGER essential oil.

    PubMed

    Ceylan, Ozgur; Ugur, Aysel

    2015-06-01

    In this study, antimicrobial and antibiofilm activities and the chemical composition of Thymus sipyleus BOISS. subsp. sipyleus BOISS. var. davisianus RONNIGER essential oil was evaluated. The essential oil was obtained by hydro-distillation and analyzed by gas chromatography-mass spectrometry. Fourteen compounds were characterized, having as major components thymol (38.31%) and carvacrol (37.95%). Minimum inhibitory concentrations (MICs) of oil and the major components were calculated by serial dilution method, and anti-biofilm effects by microplate biofilm assay against five Gram positive (Staphylococcus aureus MU 38, MU 40, MU 46, MU 47, Stahylococcus epidermidis MU 30) and five Gram negative (Pseudomonas aeruginosa MU 187, MU 188, MU 189, Pseudomonas fluorescens MU 180, MU 181) bacteria. It was found that MICs for essential oil, thymol and carvacrol were between 5 and 50 µl/ml, 0.125-0.5 µg/ml and 0.125-05 µl/ml, respectively. The results showed that doses of MIC produced a greater anti-biofilm influence than 0.5, 0.25 and 0.125 MIC. In the presence of essential oil (MIC), the mean biofilm formation value was equal to 67 ± 5.5% for P. aeruginosa MU 188, and essential oil (MIC) inhibition exceeds 60% for P. aeruginosa biofilms. The results also showed that carvacrol (MIC) was able to induce an inhibition 72.9 ± 4.1% for S.aureus (MU 40) biofilm. In addition, thymol (MIC) showed 68.6 ± 5.3% reduction in biofilm formation of P. fluorescens MU 181. This study demonstrated the antimicrobial and antibiofilm activity of T. sipyleus BOISS. subsp. sipyleus BOISS. var. davisianus RONNIGER essential oil and points out the exceptional efficiency of thymol and carvacrol, which could represent candidates in the treatment of Pseudomonas and Staphylococcus biofilms. PMID:25385321

  8. The combinations Lysobacter enzymogenes subsp. enzymogenes Christensen and Cook 1978, L. enzymogenes subsp. cookii Christensen 1978 and Streptococcus casseliflavus (Mundt and Graham 1968) Vaughan et al. 1979 were in accordance with the International Code of Nomenclature of Bacteria at the time of publication in the International Journal of Systematic Bacteriology, but are not to be considered to be included on the Approved Lists of Bacterial Names. Opinion 95: Judicial Commission of the International Committee on Systematics of Prokaryotes.

    PubMed

    Tindall, B J

    2014-11-01

    The Judicial Commission affirms that, according to information presented to it, the combination Lysobacter enzymogenes subsp. enzymogenes Christensen and Cook 1978, the combination Lysobacter enzymogenes subsp. cookii Christensen 1978 and the combination Streptococcus casseliflavus (Mundt and Graham 1968) Vaughan et al. 1979 were in accordance with the wording of the 1975 and 1992 revisions of the International Code of Nomenclature of Bacteria but they are not to be considered to be included on the Approved Lists of Bacterial Names. PMID:25386007

  9. Genome sequencing reveals a new lineage associated with lablab bean and genetic exchange between Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans

    PubMed Central

    Aritua, Valente; Harrison, James; Sapp, Melanie; Buruchara, Robin; Smith, Julian; Studholme, David J.

    2015-01-01

    Common bacterial blight is a devastating seed-borne disease of common beans that also occurs on other legume species including lablab and Lima beans. We sequenced and analyzed the genomes of 26 strains of Xanthomonas axonopodis pv. phaseoli and X. fuscans subsp. fuscans, the causative agents of this disease, collected over four decades and six continents. This revealed considerable genetic variation within both taxa, encompassing both single-nucleotide variants and differences in gene content, that could be exploited for tracking pathogen spread. The bacterial strain from Lima bean fell within the previously described Genetic Lineage 1, along with the pathovar type strain (NCPPB 3035). The strains from lablab represent a new, previously unknown genetic lineage closely related to strains of X. axonopodis pv. glycines. Finally, we identified more than 100 genes that appear to have been recently acquired by Xanthomonas axonopodis pv. phaseoli from X. fuscans subsp. fuscans. PMID:26500625

  10. Strain- and genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, a bacterial pathogen causing American foulbrood disease in honeybees.

    PubMed

    Genersch, Elke; Ashiralieva, Ainura; Fries, Ingemar

    2005-11-01

    Virulence variations of Paenibacillus larvae subsp. larvae, the causative agent of American foulbrood disease of honeybees, were investigated by analysis of 16 field isolates of this pathogen, belonging to three previously characterized genotypes, as well as the type strain (ATCC 9545) of P. larvae subsp. larvae, with exposure bioassays. We demonstrated that the strain-specific 50% lethal concentrations varied within an order of magnitude and that differences in amount of time for the pathogen to kill 100% of the infected hosts (LT100) correlated with genotype. One genotype killed rather quickly, with a mean LT100 of 7.8 +/- 1.7 days postinfection, while the other genotypes acted more slowly, with mean LT100s of 11.2 +/- 0.8 and 11.6 +/- 0.6 days postinfection. PMID:16269801

  11. Strain- and Genotype-Specific Differences in Virulence of Paenibacillus larvae subsp. larvae, a Bacterial Pathogen Causing American Foulbrood Disease in Honeybees

    PubMed Central

    Genersch, Elke; Ashiralieva, Ainura; Fries, Ingemar

    2005-01-01

    Virulence variations of Paenibacillus larvae subsp. larvae, the causative agent of American foulbrood disease of honeybees, were investigated by analysis of 16 field isolates of this pathogen, belonging to three previously characterized genotypes, as well as the type strain (ATCC 9545) of P. larvae subsp. larvae, with exposure bioassays. We demonstrated that the strain-specific 50% lethal concentrations varied within an order of magnitude and that differences in amount of time for the pathogen to kill 100% of the infected hosts (LT100) correlated with genotype. One genotype killed rather quickly, with a mean LT100 of 7.8 ± 1.7 days postinfection, while the other genotypes acted more slowly, with mean LT100s of 11.2 ± 0.8 and 11.6 ± 0.6 days postinfection. PMID:16269801

  12. Genome sequencing reveals a new lineage associated with lablab bean and genetic exchange between Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans.

    PubMed

    Aritua, Valente; Harrison, James; Sapp, Melanie; Buruchara, Robin; Smith, Julian; Studholme, David J

    2015-01-01

    Common bacterial blight is a devastating seed-borne disease of common beans that also occurs on other legume species including lablab and Lima beans. We sequenced and analyzed the genomes of 26 strains of Xanthomonas axonopodis pv. phaseoli and X. fuscans subsp. fuscans, the causative agents of this disease, collected over four decades and six continents. This revealed considerable genetic variation within both taxa, encompassing both single-nucleotide variants and differences in gene content, that could be exploited for tracking pathogen spread. The bacterial strain from Lima bean fell within the previously described Genetic Lineage 1, along with the pathovar type strain (NCPPB 3035). The strains from lablab represent a new, previously unknown genetic lineage closely related to strains of X. axonopodis pv. glycines. Finally, we identified more than 100 genes that appear to have been recently acquired by Xanthomonas axonopodis pv. phaseoli from X. fuscans subsp. fuscans. PMID:26500625

  13. Development and Validation of a Liquid Medium (M7H9C) for Routine Culture of Mycobacterium avium subsp. paratuberculosis To Replace Modified Bactec 12B Medium

    PubMed Central

    Whittington, Ann-Michele; Waldron, Anna; Begg, Douglas J.; de Silva, Kumi; Purdie, Auriol C.; Plain, Karren M.

    2013-01-01

    Liquid culture of Mycobacterium avium subsp. paratuberculosis from clinical samples, such as feces, is the most sensitive antemortem test for the diagnosis of Johne's disease in ruminants. In Australia, New Zealand, the United States, and some other countries, the Bactec 460 system with modified Bactec 12B medium (Becton, Dickinson) has been the most commonly used liquid culture system, but it was discontinued in 2012. In this study, a new liquid culture medium, M7H9C, was developed. It consists of a Middlebrook 7H9 medium base with added Casitone, albumin, dextrose, catalase, egg yolk, mycobactin J, and a cocktail of antibiotics. We found that polyoxyethylene stearate (POES) was not essential for the cultivation of M. avium subsp. paratuberculosis in either the Bactec 12B or the M7H9C medium. The limit of detection determined using pure cultures of the C and S strains of M. avium subsp. paratuberculosis was 7 bacilli per 50 ?l inoculum in the two media. The new medium was validated using 784 fecal and tissue samples from sheep and cattle, >25% of which contained viable M. avium subsp. paratuberculosis. Discrepant results for the clinical samples between the two media were mostly associated with samples that contained <10 viable bacilli per gram, but these results were relatively uncommon, and the performances of the two media were not significantly different. M7H9C medium was less than half the cost of the Bactec 12B medium and did not require regular examination during incubation, but a confirmatory IS900 PCR test had to be performed on every culture after the predetermined incubation period. PMID:24048541

  14. Simultaneous detection of Acidovorax avenae subsp. citrulli and Didymella bryoniae in cucurbit seedlots using magnetic capture hybridization and real-time polymerase chain reaction.

    PubMed

    Ha, Y; Fessehaie, A; Ling, K S; Wechter, W P; Keinath, A P; Walcott, R R

    2009-06-01

    To improve the simultaneous detection of two pathogens in cucurbit seed, a combination of magnetic capture hybridization (MCH) and multiplex real-time polymerase chain reaction (PCR) was developed. Single-stranded DNA hybridization capture probes targeting DNA of Acidovorax avenae subsp. citrulli, causal agent of bacterial fruit blotch, and Didymella bryoniae, causal agent of gummy stem blight, were covalently attached to magnetic particles and used to selectively concentrate template DNA from cucurbit seed samples. Sequestered template DNAs were subsequently amplified by multiplex real-time PCR using pathogen-specific TaqMan PCR assays. The MCH multiplex real-time PCR assay displayed a detection threshold of A. avenae subsp. citrulli at 10 CFU/ml and D. bryoniae at 10(5) conidia/ml in mixtures of pure cultures of the two pathogens, which was 10-fold more sensitive than the direct real-time PCR assays for the two pathogens separately. Although the direct real-time PCR assay displayed a detection threshold for A. avenae subsp. citrulli DNA of 100 fg/microl in 25% (1/4 samples) of the samples assayed, MCH real-time PCR demonstrated 100% detection frequency (4/4 samples) at the same DNA concentration. MCH did not improve detection sensitivity for D. bryoniae relative to direct real-time PCR using conidial suspensions or seed washes from D. bryoniae-infested cucurbit seed. However, MCH real-time PCR facilitated detection of both target pathogens in watermelon and melon seed samples (n = 5,000 seeds/sample) in which 0.02% of the seed were infested with A. avenae subsp. citrulli and 0.02% were infested with D. bryoniae. PMID:19453225

  15. Novel insights into the genomic basis of citrus canker based on the genome sequences of two strains of Xanthomonas fuscans subsp. aurantifolii

    PubMed Central

    2010-01-01

    Background Citrus canker is a disease that has severe economic impact on the citrus industry worldwide. There are three types of canker, called A, B, and C. The three types have different phenotypes and affect different citrus species. The causative agent for type A is Xanthomonas citri subsp. citri, whose genome sequence was made available in 2002. Xanthomonas fuscans subsp. aurantifolii strain B causes canker B and Xanthomonas fuscans subsp. aurantifolii strain C causes canker C. Results We have sequenced the genomes of strains B and C to draft status. We have compared their genomic content to X. citri subsp. citri and to other Xanthomonas genomes, with special emphasis on type III secreted effector repertoires. In addition to pthA, already known to be present in all three citrus canker strains, two additional effector genes, xopE3 and xopAI, are also present in all three strains and are both located on the same putative genomic island. These two effector genes, along with one other effector-like gene in the same region, are thus good candidates for being pathogenicity factors on citrus. Numerous gene content differences also exist between the three cankers strains, which can be correlated with their different virulence and host range. Particular attention was placed on the analysis of genes involved in biofilm formation and quorum sensing, type IV secretion, flagellum synthesis and motility, lipopolysacharide synthesis, and on the gene xacPNP, which codes for a natriuretic protein. Conclusion We have uncovered numerous commonalities and differences in gene content between the genomes of the pathogenic agents causing citrus canker A, B, and C and other Xanthomonas genomes. Molecular genetics can now be employed to determine the role of these genes in plant-microbe interactions. The gained knowledge will be instrumental for improving citrus canker control. PMID:20388224

  16. Spatial genetic structure in Beta vulgaris subsp. maritima and Beta macrocarpa reveals the effect of contrasting mating system, influence of marine currents, and footprints of postglacial recolonization routes

    PubMed Central

    Leys, Marie; Petit, Eric J; El-Bahloul, Yasmina; Liso, Camille; Fournet, Sylvain; Arnaud, Jean-François

    2014-01-01

    Understanding the factors that contribute to population genetic divergence across a species' range is a long-standing goal in evolutionary biology and ecological genetics. We examined the relative importance of historical and ecological features in shaping the present-day spatial patterns of genetic structure in two related plant species, Beta vulgaris subsp. maritima and Beta macrocarpa. Using nuclear and mitochondrial markers, we surveyed 93 populations from Brittany (France) to Morocco – the southern limit of their species' range distribution. Whereas B. macrocarpa showed a genotypic structure and a high level of genetic differentiation indicative of selfing, the population genetic structure of B. vulgaris subsp. maritima was consistent with an outcrossing mating system. We further showed (1) a strong geographic clustering in coastal B. vulgaris subsp. maritima populations that highlighted the influence of marine currents in shaping different lineages and (2) a peculiar genetic structure of inland B. vulgaris subsp. maritima populations that could indicate the admixture of distinct evolutionary lineages and recent expansions associated with anthropogenic disturbances. Spatial patterns of nuclear diversity and differentiation also supported a stepwise recolonization of Europe from Atlantic-Mediterranean refugia after the last glacial period, with leading-edge expansions. However, cytoplasmic diversity was not impacted by postglacial recolonization: stochastic long-distance seed dispersal mediated by major oceanic currents may mitigate the common patterns of reduced cytoplasmic diversity observed for edge populations. Overall, the patterns we documented here challenge the general view of reduced genetic diversity at the edge of a species' range distribution and provide clues for understanding how life-history and major geographic features interact to shape the distribution of genetic diversity. PMID:24963380

  17. Immunogenicity and Reactivity of Novel Mycobacterium avium subsp. paratuberculosis PPE MAP1152 and Conserved MAP1156 Proteins with Sera from Experimentally and Naturally Infected Animals ? †

    PubMed Central

    Bannantine, John P.; Paulson, Avery L.; Chacon, Ofelia; Fenton, Robert J.; Zinniel, Denise K.; McVey, David S.; Smith, David R.; Czuprynski, Charles J.; Barletta, Raúl G.

    2011-01-01

    Mycobacterium avium subsp. paratuberculosis causes Johne's disease (JD) in ruminants. Development of genetic tools and completion of the M. avium subsp. paratuberculosis genome sequencing project have expanded the opportunities for antigen discovery. In this study, we determined the seroreactivities of two proteins encoded at the 5? and 3? regions of the MAP1152-MAP1156 gene cluster. MAP1152 encodes a PPE protein, and MAP1156 encodes a diacylglycerol acyltransferase involved in triglyceride metabolism and classified in the uncharacterized protein family UPF0089. Recombinant MAP proteins were overproduced and purified from Escherichia coli as maltose-binding protein (MBP) fusions. Immunoblotting analysis indicated that both MAP1152 and MAP1156 displayed reactivity against sera of mice and rabbits immunized with live M. avium subsp. paratuberculosis cells and against samples from naturally infected cattle. In immunoblot assays, MAP1156 yielded a stronger positive signal than MAP1152 against sera from cattle with JD. An enzyme-linked immunosorbent assay for the recombinant proteins was developed and used to test preclassified positive and negative serum samples from naturally infected and noninfected cattle. Samples, with one exception, displayed no seroreactivity against the MBP-LacZ fusion protein (P > 0.05), the negative-control antigen. MAP1152 displayed seroreactivity against all positive sera but no seroreactivity to the negative sera (P < 0.01). MAP1156 displayed stronger and more variable reactivity than MAP1152, but significant differences were observed between noninfected and infected cattle (P < 0.05). Otherwise, degrees of reactivity followed the same trend as the positive reference antigen. In conclusion, both proteins are immunogenic in mice and rabbits, and M. avium subsp. paratuberculosis-infected cattle mount a humoral response to both MAP1152 and MAP1156 cross-reactive epitopes. These findings have potential applications to diagnostics, vaccine production, and elucidation of the immunopathogenesis of JD. PMID:21084462

  18. Production of buttery-odor compounds and transcriptome response in Leuconostoc gelidum subsp. gasicomitatum LMG18811T during growth on various carbon sources.

    PubMed

    Jääskeläinen, Elina; Vesterinen, Sanna; Parshintsev, Jevgeni; Johansson, Per; Riekkola, Marja-Liisa; Björkroth, Johanna

    2015-03-01

    Leuconostoc gelidum subsp. gasicomitatum is a common spoilage bacterium in meat products packaged under oxygen-containing modified atmospheres. Buttery off-odors related to diacetyl/acetoin formation are frequently associated with the spoilage of these products. A whole-genome microarray study, together with gas chromatography (GC)-mass spectrometry (MS) analyses of the pathway end products, was performed to investigate the transcriptome response of L. gelidum subsp. gasicomitatum LMG18811(T) growing on semidefined media containing glucose, ribose, or inosine, which are essential carbon sources in meat. Generally, the gene expression patterns with ribose and inosine were quite similar, indicating that catabolism of ribose and nucleosides is closely linked. Diacetyl/acetoin concentrations as high as 110 or 470 ?M were measured when growth was based on inosine or ribose, respectively. The gene expression results for pyruvate metabolism (upregulation of ?-acetolactate synthase, downregulation of l-lactate dehydrogenase and pyruvate dehydrogenase) were as expected when diacetyl and acetoin were the end products. No diacetyl production (<7.5 ?M) was detected with the glucose-containing medium, even though the cell counts of LMG18811(T) was 6 or 10 times higher than that on inosine or ribose, respectively. Although glucose was the most effective carbon source for the growth of L. gelidum subsp. gasicomitatum, utilization of inosine and ribose resulted in the production of the unwanted buttery-odor compounds. These results increase our understanding of which compounds are likely to enhance the formation of buttery odors during meat spoilage caused by L. gelidum subsp. gasicomitatum. PMID:25548057

  19. Biotransformation of aflatoxin B1 and aflatoxin G1 in peanut meal by anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus.

    PubMed

    Chen, Yujie; Kong, Qing; Chi, Chen; Shan, Shihua; Guan, Bin

    2015-10-15

    The purpose of this study was to explore the ability of anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to biotransform aflatoxins in peanut meal. The pH of the peanut meal was adjusted above 10, and then heated for 10 min at 100 °C, 115 °C and 121 °C. The S. thermophilus and L. delbrueckii subsp. bulgaricus were precultured together in MRS broth for 48 h at 37 °C. The heated peanut meal was mixed with precultured MRS broth containing 7.0×10(8) CFU/mL of S. thermophilus and 3.0×10(3) CFU/mL of L. delbrueckii subsp. bulgaricus with the ratio of 1 to 1 (weight to volume) and incubated in anaerobic jars at 37 °C for 3 days. The aflatoxin content in the peanut meal samples was determined by HPLC. The results showed that the peanut meal contained mainly aflatoxin B1 (AFB1) (10.5±0.64 ?g/kg) and aflatoxin G1 (AFG1) (18.7±0.55 ?g/kg). When heat treatment was combined with anaerobic solid fermentation, the biotransformation rate of aflatoxins in peanut meal could attain 100%. The cytotoxicity of fermented peanut meal to L929 mouse connective tissue fibroblast cells was determined by MTT assay and no significant toxicity was observed in the fermented peanut meal. Furthermore, heat treatment and anaerobic solid fermentation did not change the amino acid concentrations and profile in peanut meal. PMID:26143229

  20. Bioconversion of squid pen by Lactobacillus paracasei subsp paracasei TKU010 for the production of proteases and lettuce growth enhancing biofertilizers.

    PubMed

    Wang, San-Lang; Huang, Tsai-Yi; Wang, Chun-Yuan; Liang, Tzu-Wen; Yen, Yue-Horng; Sakata, Yusuke

    2008-09-01

    A protease-producing bacterium, strain TKU010, was isolated from infant vomited milk and identified as Lactobacillus paracasei subsp. paracasei. A surfactant-stable protease, purified 64-fold from the third day culture supernatant to homogeneity in an overall yield of 11%, has a molecular weight of about 49,000. The enzyme degraded casein and gelatin, but did not degrade albumin, fibrin, and elastin. The enzyme activity was increased about 1.5-fold by the addition of 5mM Ba(2+). However, Fe(2+) and Cu(2+) ions strongly inhibited the enzyme. The enzyme was maximally active at pH 10 and 60 degrees C and retained 94% and 71% activity in the presence of Tween 20 (2% w/v) and SDS (2mM), respectively. The result of identification of TKU010 protease showed that nine tryptic peptides were identical to Serratia protease (serralysin) (GenBank accession number gi999638) with 35% sequence coverage. In comparison with the tryptic peptides of L. paracasei subsp. paracasei TKU012 protease, TKU010 protease possessed two additional peptides with sequences of AATTGYDAVDDLLHYHER and QTFTHEIGHALGLSHPGDYNAGEGNPTYR. The fourth day culture supernatant of TKU010 showed maximal activity of about 5-fold growth enhancing effect on lettuce weight, which was not shown with L. paracasei subsp paracasei TKU012. PMID:18187322

  1. Role of glutamine synthetase adenylylation in the self-protection of Pseudomonas syringae subsp. tabaci from its toxin, tabtoxinine-. beta. -lactam

    SciTech Connect

    Knight, T.J.; Durbin, R.D.; Langston-Unkefer, P.J.

    1986-04-01

    Selected pathovars of Pseudomonas syringae produce an extracellular phytotoxin, tabtoxinine-..beta..-lactam, that irreversibly inhibits its known physiological target, glutamine synthetase (GS). Pseudomonas syringae subsp. tabaci retains significant amounts of glutamine synthetase activity during toxin production in culture. As part of our investigation of the self-protection mechanism(s) used by these pathovars, we have determined that GS becomes adenylylated after toxin production is initiated and that the serine released from the zinc-activated hydrolysis of tabtoxin is a factor in the initiation of this adenylylation. The adenylylation state of this GS was estimated to range from E/sub 5.0-7.5/. The irreversible inactivation by tabtoxinine-..beta..-lactam of unadenylylated and adenylylated glutamine synthetase purified from P. syringae subsp. tabaci was investigated. Adenylylated GS was inactivated by tabtoxinine-..beta..-lactam at a slower rate that was unadenylylated enzyme. Adenylylated GS (E/sub 7.5-10.5/) was significantly protected from this inactivation in the presence of the enzyme effectors, AMP, Ala, Gly, His, and Ser. Thus, the combination of the adenylylation of GS after toxin production is initiated and the presence of the enzyme effectors in vivo could provide part of the self-protection mechanism uses by subsp. tabaci.

  2. Duplex PCR for differential identification of Mycobacterium bovis, M. avium, and M. avium subsp. paratuberculosis in formalin- fixed paraffin-embedded tissues from cattle.

    PubMed

    Coetsier, C; Vannuffel, P; Blondeel, N; Denef, J F; Cocito, C; Gala, J L

    2000-08-01

    We previously isolated and sequenced two genomic segments of Mycobacterium avium subsp. paratuberculosis, namely, f57, a species-specific sequence, and the p34 gene, coding for a 34-kDa antigenic protein. Comparison of sequences upstream of the p34 open reading frame (us-p34) from M. avium subsp. paratuberculosis and M. tuberculosis showed a 79-base deletion in M. tuberculosis. Sequence analysis of the p34 genes in another two species, M. bovis (strain BCG) and M. avium (strain D4), confirmed the differences observed between tuberculous and nontuberculous species. A duplex diagnostic PCR strategy based on coamplification of nonhomologous us-p34 and species-specific f57 sequences was therefore developed. Duplex PCR yielded three different patterns, specific either for tuberculous bacilli (M. tuberculosis, M. bovis, and M. africanum), for both nontuberculous mycobacteria M. avium and M. intracellulare, or for M. avium subsp. paratuberculosis. The specificity of this single-step DNA-based assay was assessed on DNA from cultured mycobacterial strains, as well as on a panel of formalin-fixed and paraffin-embedded tissues from cattle. Molecular assay results from tissular DNA were compared to conventional bacteriological and histological test results, including those obtained by Ziehl-Neelsen staining on tissue biopsy specimens. Molecular discrimination was successful and confirmed the value of duplex us-p34 and f57 sequence amplification for differential diagnosis of tuberculosis, paratuberculosis, or infections caused by other members of the M. avium complex. PMID:10921976

  3. DsbA Plays a Critical and Multifaceted Role in the Production of Secreted Virulence Factors by the Phytopathogen Erwinia carotovora subsp. atroseptica*S?

    PubMed Central

    Coulthurst, Sarah J.; Lilley, Kathryn S.; Hedley, Peter E.; Liu, Hui; Toth, Ian K.; Salmond, George P. C.

    2008-01-01

    Erwinia carotovora subsp. atroseptica is an enterobacterial phytopathogen causing economically significant soft rot disease. Pathogenesis is mediated by multiple secreted virulence factors, many of which are secreted by the type II (Out) secretion system. DsbA catalyzes the introduction of disulfide bonds into periplasmic and secreted proteins. In this study, the extracellular proteome (secretome) of wild type E. carotovora subsp. atroseptica SCRI1043, and dsbA and out mutants, was analyzed by spectral counting mass spectrometry. This revealed that dsbA inactivation had a huge impact on the secretome and identified diverse DsbA- and Out-dependent secreted proteins, representing known, predicted, and novel candidate virulence factors. Further characterization of the dsbA mutant showed that secreted enzyme activities, motility, production of the quorumsensing signal, and virulence were absent or substantially reduced. The impact of DsbA on secreted virulence factor production was mediated at multiple levels, including impacting on the Out secretion system and the virulence gene regulatory network. Transcriptome analyses revealed that the abundance of a broad, but defined, set of transcripts, including many virulence factors, was altered in the dsbA mutant, identifying a new virulence regulon responsive to extracytoplasmic conditions. In conclusion, DsbA plays a crucial, multifaceted role in the pathogenesis of E. carotovora subsp. atroseptica. PMID:18562317

  4. Detection of Variants of the pRAS3, pAB5S9, and pSN254 Plasmids in Aeromonas salmonicida subsp. salmonicida: Multidrug Resistance, Interspecies Exchanges, and Plasmid Reshaping

    PubMed Central

    Vincent, Antony T.; Trudel, Mélanie V.; Paquet, Valérie E.; Boyle, Brian; Tanaka, Katherine H.; Dallaire-Dufresne, Stéphanie; Daher, Rana K.; Frenette, Michel; Derome, Nicolas

    2014-01-01

    The ubiquitous water-borne Gram-negative bacterium Aeromonas salmonicida subsp. salmonicida is the causative agent of furunculosis, a worldwide disease in fish farms. Plasmids carrying antibiotic resistance genes have already been described for this bacterium. The aim of the present study was to identify and characterize additional multidrug resistance plasmids in A. salmonicida subsp. salmonicida. We sequenced the plasmids present in two multiple antibiotic-resistant isolates using high-throughput technologies. We also investigated 19 other isolates with various multidrug resistance profiles by genotyping PCR and assessed their resistance to tetracycline. We identified variants of the pAB5S9 and pSN254 plasmids that carry several antibiotic resistance genes and that have been previously reported in bacteria other than A. salmonicida subsp. salmonicida, which suggests a high level of interspecies exchange. Genotyping analyses and the antibiotic resistance profiles of the 19 other isolates support the idea that multiple versions of pAB5S9 and pSN254 exist in A. salmonicida subsp. salmonicida. We also identified variants of the pRAS3 plasmid. The present study revealed that A. salmonicida subsp. salmonicida harbors a wide variety of plasmids, which suggests that this ubiquitous bacterium may contribute to the spread of antibiotic resistance genes in the environment. PMID:25267667

  5. Expression of Xylella fastidiosa RpfF in citrus disrupts signaling in Xanthomonas citri subsp. citri and thereby its virulence.

    PubMed

    Caserta, R; Picchi, S C; Takita, M A; Tomaz, J P; Pereira, W E L; Machado, M A; Ionescu, M; Lindow, S; De Souza, A A

    2014-11-01

    Xylella fastidiosa and Xanthomonas citri subsp. citri, that cause citrus variegated chlorosis (CVC) and citrus canker diseases, respectively, utilize diffusible signal factor (DSF) for quorum sensing. DSF, produced by RpfF, are similar fatty acids in both organisms, although a different set of genes is regulated by DSF in each species. Because of this similarity, Xylella fastidiosa DSF might be recognized and affect the biology of Xanthomonas citri. Therefore, transgenic Citrus sinensis and Carrizo citrange plants overexpressing the Xylella fastidiosa rpfF were inoculated with Xanthomonas citri and changes in symptoms of citrus canker were observed. X. citri biofilms formed only at wound sites on transgenic leaves and were thicker; however, bacteria were unable to break through the tissue and form pustules elsewhere. Although abundant growth of X. citri occurred at wound sites on inoculated transgenic leaves, little growth was observed on unwounded tissue. Genes in the DFS-responsive core in X. citri were downregulated in bacteria isolated from transgenic leaves. DSF-dependent expression of engA was suppressed in cells exposed to xylem sap from transgenic plants. Thus, altered symptom development appears to be due to reduced expression of virulence genes because of the presence of antagonists of DSF signaling in X. citri in rpfF-expressing plants. PMID:25099341

  6. Survey on the presence of Mycobacterium avium subsp. paratuberculosis in ground beef from an industrial meat plant.

    PubMed

    Savi, R; Ricchi, M; Cammi, G; Garbarino, C; Leo, S; Pongolini, S; Arrigoni, N

    2015-06-12

    Paratuberculosis of ruminants is characterised by chronic enteritis but, at advanced stages of the disease, a systemic dissemination of Mycobacterium avium subsp. paratuberculosis (MAP) in tissues and organs can occur. MAP has been recovered from lymph nodes and muscles of clinical and sub-clinical cows. In most countries, dairy and beef cattle infected with paratuberculosis are routinely sent to slaughter and the consumption of their meat could be a possible route of human exposure to MAP. However, few studies on MAP in ground beef are currently available. During the period November 2013-March 2014 we carried out a survey on the ground beef produced in an industrial meat processing plant. One-hundred and forty samples of ground meat were analysed by IS900-qPCR and culture (VersaTrek System). The limit of detection (LOD) of qPCR was 630 MAP cells/g (107 CFU/g) while the LOD for culture was 170-230 MAP cells/g (62-115 CFU/g). No samples were positive by direct IS900 qPCR, while two samples were positive by liquid culture. Our data suggest that the presence of live MAP in raw minced meat is possible. In order to avoid exposure for humans through the consumption of contaminated meat, proper cooking of meat is recommended. PMID:25824129

  7. Effect of lipoarabinomannan from Mycobacterium avium subsp avium in Freund's incomplete adjuvant on the immune response of cattle

    PubMed Central

    Colavecchia, S.B.; Jolly, A.; Fernández, B.; Fontanals, A.M.; Fernández, E.; Mundo, S.L.

    2012-01-01

    The aim of the present study was to determine whether lipoarabinomannan (LAM), in combination with Freund's incomplete adjuvant (FIA), was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA) in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-? production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-? production in response to ConA and OVA (P < 0.05). Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-? production by at least 50 and 25%, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis. PMID:22286534

  8. Sequence diversity of Treponema pallidum subsp. pallidum tprK in human syphilis lesions and rabbit-propagated isolates.

    PubMed

    LaFond, Rebecca E; Centurion-Lara, Arturo; Godornes, Charmie; Rompalo, Anne M; Van Voorhis, Wesley C; Lukehart, Sheila A

    2003-11-01

    The tprK gene of Treponema pallidum subsp. pallidum, the causative agent of venereal syphilis, belongs to a 12-member gene family and encodes a protein with a predicted cleavable signal sequence and predicted transmembrane domains. Except for the Nichols type strain, all rabbit-propagated isolates of T. pallidum examined thus far are comprised of mixed populations of organisms with heterogeneous tprK sequences. We show that tprK sequences in treponemes obtained directly from syphilis patients are also heterogeneous. Clustering analysis demonstrates that primary chancre tprK sequences are more likely to cluster within a sample than among samples and that tighter clustering is seen within chancre samples than within rabbit-propagated isolates. Closer analysis of tprK sequences from a rabbit-propagated isolate reveals that individual variable regions have different levels of diversity, suggesting that variable regions may have different intrinsic rates of sequence change or may be under different levels of selection. Most variable regions show increased sequence diversity upon passage. We speculate that the diversification of tprK during infection allows organisms to evade the host immune response, contributing to reinfection and persistent infection. PMID:14563860

  9. Genome-wide analysis of the SBP-box gene family in Chinese cabbage (Brassica rapa subsp. pekinensis).

    PubMed

    Tan, Hua-Wei; Song, Xiao-Ming; Duan, Wei-Ke; Wang, Yan; Hou, Xi-Lin

    2015-11-01

    The SQUAMOSA PROMOTER BINDING PROTEIN (SBP)-box gene family contains highly conserved plant-specific transcription factors that play an important role in plant development, especially in flowering. Chinese cabbage (Brassica rapa subsp. pekinensis) is a leafy vegetable grown worldwide and is used as a model crop for research in genome duplication. The present study aimed to characterize the SBP-box transcription factor genes in Chinese cabbage. Twenty-nine SBP-box genes were identified in the Chinese cabbage genome and classified into six groups. We identified 23 orthologous and 5 co-orthologous SBP-box gene pairs between Chines cabbage and Arabidopsis. An interaction network among these genes was constructed. Sixteen SBP-box genes were expressed more abundantly in flowers than in other tissues, suggesting their involvement in flowering. We show that the MiR156/157 family members may regulate the coding regions or 3'-UTR regions of Chinese cabbage SBP-box genes. As SBP-box genes were found to potentially participate in some plant development pathways, quantitative real-time PCR analysis was performed and showed that Chinese cabbage SBP-box genes were also sensitive to the exogenous hormones methyl jasmonic acid and salicylic acid. The SBP-box genes have undergone gene duplication and loss, evolving a more refined regulation for diverse stimulation in plant tissues. Our comprehensive genome-wide analysis provides insights into the SBP-box gene family of Chinese cabbage. PMID:26599708

  10. Morphology, Carbohydrate Composition and Vernalization Response in a Genetically Diverse Collection of Asian and European Turnips (Brassica rapa subsp. rapa)

    PubMed Central

    Zhang, Ningwen; Zhao, Jianjun; Lens, Frederic; de Visser, Joan; Menamo, Temesgen; Fang, Wen; Xiao, Dong; Bucher, Johan; Basnet, Ram Kumar; Lin, Ke; Cheng, Feng; Wang, Xiaowu; Bonnema, Guusje

    2014-01-01

    Brassica rapa displays enormous morphological diversity, with leafy vegetables, turnips and oil crops. Turnips (Brassica rapa subsp. rapa) represent one of the morphotypes, which form tubers and can be used to study the genetics underlying storage organ formation. In the present study we investigated several characteristics of an extensive turnip collection comprising 56 accessions from both Asia (mainly Japanese origin) and Europe. Population structure was calculated using data from 280 evenly distributed SNP markers over 56 turnip accessions. We studied the anatomy of turnip tubers and measured carbohydrate composition of the mature turnip tubers of a subset of the collection. The variation in 16 leaf traits, 12 tuber traits and flowering time was evaluated in five independent experiments for the entire collection. The effect of vernalization on flowering and tuber formation was also investigated. SNP marker profiling basically divided the turnip accessions into two subpopulations, with admixture, generally corresponding with geographical origin (Europe or Asia). The enlarged turnip tuber consists of both hypocotyl and root tissue, but the proportion of the two tissues differs between accessions. The ratio of sucrose to fructose and glucose differed among accessions, while generally starch content was low. The evaluated traits segregated in both subpopulations, with leaf shape, tuber colour and number of shoots per tuber explaining most variation between the two subpopulations. Vernalization resulted in reduced flowering time and smaller tubers for the Asian turnips whereas the European turnips were less affected by vernalization. PMID:25474111

  11. Effect of X-irradiation on Citrus Canker Pathogen Xanthomonas citri subsp. citri of Satsuma Mandarin Fruits.

    PubMed

    Song, Min-A; Park, Jae Sin; Kim, Ki Deok; Jeun, Yong Chull

    2015-12-01

    Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most important bacterial diseases of citrus. Because citrus canker is not found in many countries including European Union and Australia, Xcc is strictly regulated in order to prevent its spread. In this study, the effects of X-irradiation on Xcc growth either in the suspension or on the surface of citrus fruits were investigated. The suspension containing 1×10(7) cfu/ml of Xcc was irradiated with different absorbed doses of X-irradiation ranging from 50 to 400 Gy. The results showed that Xcc was fully dead at 400 Gy of X-irradiation. To determine the effect of X-irradiation on quarantine, the Xcc-inoculated citrus fruits were irradiated with different X-ray doses at which Xcc was completely inhibited by an irradiation dose of 250 Gy. The D10 value for Xcc on citrus fruits was found to be 97 Gy, indicating the possibility of direct application on citrus quarantine without any side sterilizer. Beside, presence of Xcc on the surface of asymptomatic citrus fruits obtained from citrus canker-infected orchards was noted. It indicated that the exporting citrus fruits need any treatment so that Xcc on the citrus fruits should be completely eliminated. Based on these results, ionizing radiation can be considered as an alternative method of eradicating Xcc for export of citrus fruits. PMID:26672670

  12. ERIC-PCR genotyping of field isolates of Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum.

    PubMed

    Secundo de Souza, Andrei Itajahy; Freitas Neto, Oliveiro Caetano de; Batista, Diego Felipe Alves; Estupinan, Anny Lucia Del Pilar Celis; Almeida, Adriana Maria de; Barrow, Paul Andrew; Berchieri, Angelo

    2015-12-01

    Salmonella Gallinarum (SG) and Salmonella Pullorum (SP) have been classified as biovars belonging to Salmonella enterica subsp. enterica serovar Gallinarum. Genetic diversity among isolates of the same biovar can be detected by DNA fingerprinting techniques which are useful in epidemiological investigations. In this study, we applied the PCR amplification of Enterobacterial Repetitive Intergenic Consensus sequences (ERIC-PCR) to analyse 45 strains of SG and SP, most of which were isolated from diseased poultry of different Brazilian regions over a period of 27 years until 2014. The ERIC-genotypes obtained were used to describe the epidemiological relationship amongst the strains. Our findings showed that there were six ERIC-patterns for SG strains at 80% similarity. In addition, some of the SG isolates recovered from different regions and years clustered with 100% similarity, suggesting that transfer of genotypes between these regions has taken place. The commercial rough vaccine strain 9R showed a unique profile. Meanwhile, more genetic diversity was observed among SP strains where ten ERIC-patterns were also formed at 80% similarity. PMID:26365161

  13. CRISPR-MVLST subtyping of Salmonella enterica subsp. enterica serovars Typhimurium and Heidelberg and application in identifying outbreak isolates

    PubMed Central

    2013-01-01

    Background Salmonella enterica subsp. enterica serovars Typhimurium (S. Typhimurium) and Heidelberg (S. Heidelberg) are major causes of foodborne salmonellosis, accounting for a fifth of all annual salmonellosis cases in the United States. Rapid, efficient and accurate methods for identification are required for routine surveillance and to track specific strains during outbreaks. We used Pulsed-field Gel Electrophoresis (PFGE) and a recently developed molecular subtyping approach termed CRISPR-MVLST that exploits the hypervariable nature of virulence genes and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) to subtype clinical S. Typhimurium and S. Heidelberg isolates. Results We analyzed a broad set of 175 S. Heidelberg and S. Typhimurium isolates collected over a five-year period. We identified 21 Heidelberg Sequence Types (HSTs) and 37 Typhimurium STs (TSTs) that were represented by 27 and 45 PFGE pulsotypes, respectively, and determined the discriminatory power of each method. Conclusions For S. Heidelberg, our data shows that combined typing by both CRISPR-MVLST and PFGE provided a discriminatory power of 0.9213. Importantly, CRISPR-MVLST was able to separate common PFGE patterns such as JF6X01.0022 into distinct STs, thus providing significantly greater discriminatory power. Conversely, we show that subtyping by either CRISPR-MVLST or PFGE independently provides a sufficient discriminatory power (0.9345 and 0.9456, respectively) for S. Typhimurium. Additionally, using isolates from two S. Typhimurium outbreaks, we demonstrate that CRISPR-MVLST provides excellent epidemiologic concordance. PMID:24219629

  14. Development of a predictive model for detection of Mycobacterium avium subsp. paratuberculosis in faeces by quantitative real time PCR.

    PubMed

    Kralik, Petr; Slana, Iva; Kralova, Alena; Babak, Vladimir; Whitlock, Robert H; Pavlik, Ivo

    2011-04-21

    This study focused on the development of a reliable and cost-efficient DNA isolation procedure for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in faeces by previously developed IS900 and F57 quantitative real time PCR (qPCR) and their comparison with culture. The recovery of MAP DNA from the spiking experiments ranged from 29.1 to 102.4% of the input amount of MAP with median 37.9%. The limit of detection was determined to be 1.03 × 10(4) for F57 qPCR and 6.87 × 10(2)MAP cells per gram of faeces for IS900 qPCR, respectively. The developed technique for DNA isolation was coupled with IS900 qPCR and compared to traditional MAP culture using a cohort of 1906 faecal samples examined from 12 dairy cattle farms in our laboratory. From those 1906 original faecal samples, 875 were positive by IS900 qPCR and 169 by culture. None of the culture positive samples was negative by IS900 qPCR. This data facilitated development of a predictive model capable of estimating the probability of being culture positive by estimating the absolute number of MAP per gram of faeces as determined IS900 qPCR without performing the culture. PMID:21075565

  15. First isolation of mycobacterium avium subsp. Paratuberculosis from wild guanacos (Lama guanicoe) on Tierra del Fuego Island.

    PubMed

    Salgado, M; Herthnek, D; Bölske, G; Leiva, S; Kruze, J

    2009-04-01

    The aim of this study was to search for Mycobacterium avium subsp. paratuberculosis (Map) infection in a free-ranging wild animal species in a region where Johnes's disease has yet to be reported and to classify Map isolates using a genomic typing method. Fecal samples were obtained from 501 wild guanacos (Lama guanicoe) from Tierra del Fuego Island, Chile, in August 2006. Samples were cultured using Herrold's egg yolk medium with and without mycobactin J. After 9 mo of incubation, suspected Map colonies showing mycobactin dependence were confirmed by real-time polymerase chain reaction (PCR) based on IS900 and F57. Isolates were further tested using IS1311 PCR with restriction endonuclease analysis in order to type the guanaco Map strains. Twenty-one of 501 (4.2%) animals were fecal culture-positive for Map; identity was confirmed by real-time PCR and isolates were classified as cattle-type. Most culture-positive animals were located in four contiguous geographic areas, and the infection was most commonly found among adult animals. Prevalence was higher in females (5.9%) than males (3.1%) but the difference was not statistically significant. This represents the first isolation of Map from a free-ranging wildlife species in Chile. It expands the geographic range of paratuberculosis and the diversity of wildlife species that can become infected with Map. PMID:19395739

  16. Mycobacterium avium subsp. paratuberculosis viability determination using F57 quantitative PCR in combination with propidium monoazide treatment.

    PubMed

    Kralik, P; Nocker, A; Pavlik, I

    2010-07-31

    Mycobacterium avium subsp. paratuberculosis (MAP) is known to be a very slow-growing organism. The fact that cells typically need several weeks to form visible colonies severely compromises the suitability of plate counting for assessment of viable cell counts. This problem might be overcome by the application of fast molecular methods containing a viability component. We have evaluated a promising technology combining sample treatment with propidium monoazide (PMA) prior to DNA extraction for selective detection of cells with intact cell membranes with detection of sequence element F57 by quantitative PCR (F57 qPCR). Element F57 is unique for MAP and is not known to exist in any other bacterial species. Conditions of PMA treatment were optimised for MAP isolate 7082 using live and heat-killed cells and comparing different DNA extraction procedures. The subsequent successful application of the optimised protocol to four other MAP isolates of different origins suggested that the optimised protocol might be broadly applicable to different MAP strains. Furthermore, different equations were compared to use the data resulting from this technology to optimally predict the percentage of live MAP cells in mixtures containing both live and dead cells. The presented protocol holds promise to be used routinely for detecting MAP with intact cell membranes in research applications. PMID:20385417

  17. Anti-Inflammatory and Cytostatic Activities of a Parthenolide-Like Sesquiterpene Lactone from Cota palaestina subsp. syriaca

    PubMed Central

    Talhouk, Rabih S.; Nasr, Bilal; Fares, Mohamed-Bilal; Ajeeb, Bushra; Nahhas, Rana; Al Aaraj, Lamis; Ghaddar, Tarek H.; Saliba, Najat A.

    2015-01-01

    A sesquiterpene lactone 1-?,10-Epoxy-6-hydroxy-1,10H-inunolide (K100) was isolated through “bioassay-guided fractionation” from Cota palaestina subsp. syriaca, an Eastern Mediterranean endemic plant. K100 inhibited endotoxin- (ET-) induced proinflammatory markers: IL-6, MMP-9, and NO in normal mouse mammary SCp2 Cells. Molecular docking in silico suggested that K100, having highly analogous structure as parthenolide (PTL), an anticancer compound, could bind PTL target proteins at similar positions and with comparable binding affinities. Both compounds, K100 and PTL, inhibited the proliferation and prolonged the S-phase of the cell cycle of breast adenocarcinoma MDA-MB-231 cells grown in 2D cultures. Noncytotoxic concentrations of K100 and PTL decreased the proliferation rate of MDA-MB-231 and shifted their morphology from stellate to spherical colonies in 3D cultures. This was accompanied with a significant increase in the amount of small colonies and a decrease in the amount of large colonies. Moreover, K100 and PTL decreased cellular motility and invasiveness of MDA-MB-231 cells. In summary, these results suggest that K100 exhibits PTL-analogous anti-inflammatory, cytostatic, and antimetastatic effects. PMID:26089941

  18. Evolutionary optimization of life-history traits in the sea beet Beta vulgaris subsp. maritima: Comparing model to data

    NASA Astrophysics Data System (ADS)

    Hautekèete, N.-C.; Van Dijk, H.; Piquot, Y.; Teriokhin, A.

    2009-01-01

    At evolutionary equilibrium, ecological factors will determine the optimal combination of life-history trait values of an organism. This optimum can be assessed by assuming that the species maximizes some criterion of fitness such as the Malthusian coefficient or lifetime reproductive success depending on the degree of density-dependence. We investigated the impact of the amount of resources and habitat stability on a plant's age at maturity and life span by using an evolutionary optimization model in combination with empirical data. We conducted this study on sea beet, Beta vulgaris subsp. maritima, because of its large variation in life span and age at first reproduction along a latitudinal gradient including considerable ecological variation. We also compared the consequence in our evolutionary model of maximizing either the Malthusian coefficient or the lifetime reproductive success. Both the data analysis and the results of evolutionary modeling pointed to habitat disturbance and resources like length of the growing season as factors negatively related to life span and age at maturity in sea beet. Resource availability had a negative theoretical influence with the Malthusian coefficient as the chosen optimality criterion, while there was no influence in the case of lifetime reproductive success. As suggested by previous theoretical work the final conclusion on what criterion is more adequate depends on the assumptions of how in reality density-dependence restrains population growth. In our case of sea beet data R0 seems to be less appropriate than ?.

  19. Bifidobacterium longum subsp. infantis in experimental necrotizing enterocolitis: alterations in inflammation, innate immune response, and the microbiota

    PubMed Central

    Underwood, Mark A.; Arriola, Jennifer; Gerber, Colin W.; Kaveti, Ashwini; Kalanetra, Karen M.; Kananurak, Anchasa; Bevins, Charles L.; Mills, David A.; Dvorak, Bohuslav

    2014-01-01

    Background Probiotics decrease the risk of necrotizing enterocolits (NEC). We sought to determine the impact of Bifidobacterium longum subsp. infantis (B. infantis) in the established rat model of NEC. Methods Rat pups delivered one day prior to term gestation were assigned to one of three groups: dam-fed (DF), formula-fed (FF), or fed with formula supplemented with 5 × 106 CFU B. infantis per day (FF+Binf). Experimental pups were exposed to hypoxia and cold stress. Ileal tissue was examined for pathology and expression of inflammatory mediators, antimicrobial peptides, and goblet-cell products. Ceca were assessed for bacterial composition by analysis of 16S rRNA sequence. Results Administration of B. infantis significantly reduced the incidence of NEC, decreased expression of Il6, Cxcl1, Tnfa, Il23, and iNOS, and decreased expression of the antimicrobial peptides Reg3b and Reg3g. There was significant microbial heterogeneity both within groups and between experiments. The cecal microbiota was not significantly different between the FF and FF+Binf groups. Bifidobacteria were not detected in the cecum in significant numbers. Conclusions In the rat model, the inflammation associated with NEC was attenuated by administration of probiotic B. infantis. Dysbiosis was highly variable precluding determination of the precise role of the microbiota in experimental NEC. PMID:25000347

  20. The use of date waste for lactic acid production by a fed-batch culture using Lactobacillus casei subsp. rhamnosus

    PubMed Central

    Nancib, Aicha; Nancib, Nabil; Boubendir, Abdelhafid; Boudrant, Joseph

    2015-01-01

    The production of lactic acid from date juice by Lactobacillus caseisubsp. rhamnosus in batch and fed-batch cultures has been investigated. The fed-batch culture system gave better results for lactic acid production and volumetric productivity. The aim of this work is to determine the effects of the feeding rate and the concentration of the feeding medium containing date juice glucose on the cell growth, the consumption of glucose and the lactic acid production by Lactobacillus casei subsp. rhamnosus in fed-batch cultures. For this study, two concentrations of the feeding medium (62 and 100 g/L of date juice glucose) were tested at different feeding rates (18, 22, 33, 75 and 150 mL/h). The highest volumetric productivity (1.3 g/L.h) and lactic acid yield (1.7 g/g) were obtained at a feeding rate of 33 mL/h and a date juice glucose concentration of 62 g/L in the feeding medium. As a result, most of the date juice glucose was completely utilised (residual glucose 1 g/L), and a maximum lactic acid production level (89.2 g/L) was obtained. PMID:26413076