240 research outputs found

    Thermal Stress Responses of \u3cem\u3eSodalis Glossinidius\u3c/em\u3e, an Indigenous Bacterial Symbiont of Hematophagous Tsetse Flies

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    Tsetse flies (Diptera: Glossinidae) house a taxonomically diverse microbiota that includes environmentally acquired bacteria, maternally transmitted symbiotic bacteria, and pathogenic African trypanosomes. Sodalis glossinidius, which is a facultative symbiont that resides intra and extracellularly within multiple tsetse tissues, has been implicated as a mediator of trypanosome infection establishment in the flyā€™s gut. Tsetseā€™s gut-associated population of Sodalis are subjected to marked temperature fluctuations each time their ectothermic fly host imbibes vertebrate blood. The molecular mechanisms that Sodalis employs to deal with this heat stress are unknown. In this study, we examined the thermal tolerance and heat shock response of Sodalis. When grown on BHI agar plates, the bacterium exhibited the most prolific growth at 25oC, and did not grow at temperatures above 30oC. Growth on BHI agar plates at 31Ā°C was dependent on either the addition of blood to the agar or reduction in oxygen levels. Sodalis was viable in liquid cultures for 24 hours at 30oC, but began to die upon further exposure. The rate of death increased with increased temperature. Similarly, Sodalis was able to survive for 48 hours within tsetse flies housed at 30oC, while a higher temperature (37oC) was lethal. Sodalisā€™ genome contains homologues of the heat shock chaperone protein-encoding genes dnaK, dnaJ, and grpE, and their expression was up-regulated in thermally stressed Sodalis, both in vitro and in vivo within tsetse fly midguts. Arrested growth of E. coli dnaK, dnaJ, or grpE mutants under thermal stress was reversed when the cells were transformed with a low copy plasmid that encoded the Sodalis homologues of these genes. The information contained in this study provides insight into how arthropod vector enteric commensals, many of which mediate their hostā€™s ability to transmit pathogens, mitigate heat shock associated with the ingestion of a blood meal

    Permeability-porosity relationships in seafloor vent deposits : dependence on pore evolution processes

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    Author Posting. Ā© American Geophysical Union, 2007. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 112 (2007): B05208, doi:10.1029/2006JB004716.Systematic laboratory measurements of permeability and porosity were conducted on three large vent structures from the Mothra Hydrothermal vent field on the Endeavor segment of the Juan de Fuca Ridge. Geometric means of permeability values obtained from a probe permeameter are 5.9 Ɨ 10āˆ’15 m2 for Phang, a tall sulfide-dominated spire that was not actively venting when sampled; 1.4 Ɨ 10āˆ’14 m2 for Roane, a lower-temperature spire with dense macrofaunal communities growing on its sides that was venting diffuse fluid of <300Ā°C; and 1.6 Ɨ 10āˆ’14 m2 for Finn, an active black smoker with a well-defined inner conduit that was venting 302Ā°C fluids prior to recovery. Twenty-three cylindrical cores were then taken from these vent structures. Permeability and porosity of the drill cores were determined on the basis of Darcy's law and Boyle's law, respectively. Permeability values range from āˆ¼10āˆ’15 to 10āˆ’13 m2 for core samples from Phang, from āˆ¼10āˆ’15 to 10āˆ’12 m2 for cores from Roane, and from āˆ¼10āˆ’15 to 3 Ɨ 10āˆ’13 m2 for cores from Finn, in good agreement with the probe permeability measurements. Permeability and porosity relationships are best described by two different power law relationships with exponents of āˆ¼9 (group I) and āˆ¼3 (group II). Microstructural analyses reveal that the difference in the two permeability-porosity relationships reflects different mineral precipitation processes as pore space evolves within different parts of the vent structures, either with angular sulfide grains depositing as aggregates that block fluid paths very efficiently (group I), or by late stage amorphous silica that coats existing grains and reduces fluid paths more gradually (group II). The results suggest that quantification of permeability and porosity relationships leads to a better understanding of pore evolution processes. Correctly identifying permeability and porosity relationships is an important first step toward accurately estimating fluid distribution, flow rate, and environmental conditions within seafloor vent deposits, which has important consequences for chimney growth and biological communities that reside within and on vent structures.Support from the National Science Foundation under grants NSF OCE-9986456 (W.Z. and M.K.T.) and NSF OCE-0327488 (P.R.C.) is gratefully acknowledged. We also thank the WHOI summer student fellowship for providing support to H.G

    Light at deep sea hydrothermal vents

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    Elucidation of a novel Vibrio cholerae lipid A secondary hydroxy-acyltransferase and its role in innate immune recognition

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    Similar to most Gram-negative bacteria, the outer leaflet of the outer membrane of Vibrio cholerae is comprised of lipopolysaccharide. Previous reports have proposed that V. cholerae serogroups O1 and O139 synthesize structurally different lipid A domains, which anchor lipopolysaccharide within the outer membrane. In the current study, intact lipid A species of V. cholerae O1 and O139 were analysed by mass spectrometry. We demonstrate that V. cholerae serogroups associated with human disease synthesize a similar asymmetrical hexa-acylated lipid A species, bearing a myristate (C14:0) and 3-hydroxylaurate (3-OH C12:0) at the 2ā€²- and 3ā€²-positions respectively. A previous report from our laboratory characterized the V. cholerae LpxL homologue Vc0213, which transfers a C14:0 to the 2ā€²-position of the glucosamine disaccharide. Our current findings identify V. cholerae Vc0212 as a novel lipid A secondary hydroxy-acyltransferase, termed LpxN, responsible for transferring the 3-hydroxylaurate (3-OH C12:0) to the V. cholerae lipid A domain. Importantly, the presence of a 3-hydroxyl group on the 3ā€²-linked secondary acyl chain was found to promote antimicrobial peptide resistance in V. cholerae; however, this functional group was not required for activation of the innate immune response

    The expression of plasmid mediated afimbrial adhesin genes in an avian septicemic Escherichia coli strain

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    An Escherichia coli strain (SEPT13) isolated from the liver of a hen presenting clinical signs of septicaemia had a LD50 of 4.0 Ɨ 105 CFU/ml in one-day-old chickens, expressed Ia, Ib, E1, E3, K and B colicins and aerobactin. The strain was ampicillin and streptomycin resistant, and found to have fimA, csgA and tsh DNA related sequences; it could adhere to and invade HEp-2 and tracheal epithelial cells, expressed fimbriae (observed by electron microscopy), and had five plasmids of 2.7, 4.7, 43, 56, and 88 MDa. Transposon mutagenesis of strain SEPT13, with transposon TnphoA, resulted in a mutant strain named ST16 that had a LD50 of 1.2 Ɨ 1012 CFU/ml. All other biological characteristics of strain ST16 were the same as those detected for strain SEPT13 except for the migration of an 88 MDa plasmid to the 93 MDa position indicating the insertion of the transposon into the 88 MDa plasmid. The 93 MDa plasmid of strain ST16 was transferred, by electroporation assay, to non-pathogenic receptor strains (E. coli strains K12 MS101 and HB101), resulting in transformant strains A and B, respectively. These strains exhibited adhesion properties to in vitro cultivated HEp-2 cells but did not have the capacity for invasion. The adherence occurred despite the absence of fimbriae; this finding suggests that the 88 MDa plasmid has afimbrial adhesin genes

    Geochemistry of abyssal peridotites (Mid-Atlantic Ridge, 15Ā°20ā€²N, ODP Leg 209) : implications for fluid/rock interaction in slow spreading environments

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    Author Posting. Ā© Elsevier B.V., 2006. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Chemical Geology 234 (2006): 179-210, doi:10.1016/j.chemgeo.2006.04.011.Abyssal peridotite from the 15Ā°20ā€™N area of the Mid-Atlantic Ridge show complex geochemical variations among the different sites drilled during ODP Leg 209. Major element compositions indicate variable degrees of melt depletion and refertilization as well as local hydrothermal metasomatism. Strongest evidence for melt-rock interactions are correlated Light Rare Earth Element (LREE) and High Field Strength Element (HFSE) additions at sites 1270 and 1271. In contrast, hydrothermal alteration at Sites 1274, 1272, and 1268 causes LREE mobility associated with minor HFSE variability, reflecting the low solubility of HFSE in aqueous solutions. Site 1274 contains the least-altered, highly refractory, peridotite with strong depletion in LREE and shows a gradual increase in the intensity of isochemical serpentinization; except for the addition of H2O which causes a mass gain of up to 20 g/100 g. The formation of magnetite is reflected in decreasing Fe2+/Fe3+ ratios. This style of alteration is referred to as rock-dominated serpentinization. In contrast, fluid-dominated serpentinization at Site 1268 is characterized by gains in sulfur and development of U-shaped REE pattern with strong positive Eu anomalies which are also characteristic for hot (350 to 400Ā°C) vent-type fluids discharging from black smoker fields. Serpentinites at Site 1268 were overprinted by talc alteration under static conditions due to interaction with high aSiO2 fluids causing the development of smooth, LREE-enriched patterns with pronounced negative Eu anomalies. These results show that hydrothermal fluid-peridotite and fluid-serpentinite interaction processes are an important factor regarding the budget of exchange processes between the lithosphere and the hydrosphere in slow spreading environments.ODP is sponsored by the U.S. National Science Foundation (NSF) and participating countries under management of Joint Oceanographic Institutions (JOI), Inc

    S-33 constraints on the seawater sulfate contribution in modern seafloor hydrothermal vent sulfides

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    Author Posting. Ā© Elsevier B.V., 2006. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Geochimica et Cosmochimica Acta 71 (2007): 1170-1182, doi:10.1016/j.gca.2006.11.017.Sulfide sulfur in mid-oceanic ridge hydrothermal vents is derived from leaching of basaltic-sulfide and seawater-derived sulfate that is reduced during high temperature water rock interaction. Conventional sulfur isotope studies, however, are inconclusive about the mass-balance between the two sources because 34S/32S ratios of vent fluid H2S and chimney sulfide minerals may reflect not only the mixing ratio but also isotope exchange between sulfate and sulfide. Here, we show that high-precision analysis of S-33 can provide a unique constraint because isotope mixing and isotope exchange result in different Ī”33S (ā‰” Ī“33S ā€“ 0.515 Ī“34S) values of up to 0.04 ā€° even if Ī“34S values are identical. Detection of such small Ī”33S differences is technically feasible by using the SF6 dual-inlet mass-spectrometry protocol that has been improved to achieve a precision as good as 0.006 ā€° (2Ļƒ). Sulfide minerals (marcasite, pyrite, chalcopyrite, and sphalerite) and vent H2S collected from four active seafloor hydrothermal vent sites, East Pacific Rise (EPR) 9-10Ā° N, 13Ā° N, and 21Ā° S and Mid-Atlantic Ridge (MAR) 37Ā° N yield Ī”33S values ranging from ā€“0.002 to 0.033 and Ī“34S from ā€“0.5 to 5.3 ā€°. The combined Ī“34S and Ī”33S systematics reveal that 73 to 89 % of vent sulfides are derived from leaching from basaltic sulfide and only 11 to 27 % from seawater-derived sulfate. Pyrite from EPR 13Ā° N and marcasite from MAR 37Ā° N are in isotope disequilibrium not only in Ī“34S but also in Ī”33S with respect to associated sphalerite and chalcopyrite, suggesting non-equilibrium sulfur isotope exchange between seawater sulfate and sulfide during pyrite precipitation. Seafloor hydrothermal vent sulfides are characterized by low Ī”33S values compared with biogenic sulfides, suggesting little or no contribution of sulfide from microbial sulfate reduction into hydrothermal sulfides at sediment-free mid-oceanic ridge systems. We conclude that 33S is an effective new tracer for interplay among seawater, oceanic crust and microbes in subseafloor hydrothermal sulfur cycles.S. Ono thanks the Agouron Institute for financial support and funding from the NASA Astrobiology Institute and Carnegie Institution of Washington for supporting the analytical costs. Funding for O. Rouxel is from the Deep Ocean Exploration Institute at WHOI
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