Excretion of Fumonisin B1, Hydrolyzed Fumonisin B1, and the Fumonisin B1−Fructose Adduct in Rats

The excretion of fumonisin B1 (FB1), hydrolyzed FB1 (HFB1), and FB1−fructose addition products (FB1−fructose) was determined in male Fisher 344/NHsd rats. Rats were dosed by gavage with 0.69, 6.93, or 69.3 μmol/kg of body weight FB1, HFB1, or FB1−fructose. Excretion of unchanged FB1, HFB1, and HFB1 after hydrolysis was determined in urine and feces by analytical reverse phase HPLC and fluorometric detection of the o-phthaldialdehyde derivatives. Average total FB1 backbone excretion in feces was 101, 76, and 50% of the dose for FB1, HFB1, and FB1−fructose, respectively. No effect of dose level was found on the percentage of the dose excreted as total FB1 after hydrolysis. FB1−fructose appears to be absorbed to the highest extent, followed by HFB1. FB1 appears to be excreted nearly completely in the feces. The greater absorption of HFB1 may explain the greater toxicity of HFB1 compared to FB1 on a molar basis. However, the detoxification of FB1 by formation of the fructose adduct cannot be explained by reduced absorption. Average total FB1 backbone excretion in urine was 2.7, 5.0, and 5.3% of the dose for FB1, HFB1, or FB1−fructose, respectively

Tetraether membrane lipids of Candidatus “Aciduliprofundum boonei”, a cultivated obligate thermoacidophilic euryarchaeote from deep-sea hydrothermal vents

The lipid composition of Candidatus “Aciduliprofundum boonei”, the only cultivated representative of archaea falling in the DHVE2 phylogenetic cluster, a group of microorganisms ubiquitously occurring at hydrothermal vents, was studied. The predominant core membrane lipids in this thermophilic euryarchaeote were found to be composed of glycerol dibiphytanyl glycerol tetraethers (GDGTs) containing 0–4 cyclopentyl moieties. In addition, GDGTs with an additional covalent bond between the isoprenoid hydrocarbon chains, so-called H-shaped GDGTs, were present. The latter core lipids have been rarely reported previously. Intact polar lipid analysis revealed that they predominantly consist of GDGTs with a phospho-glycerol headgroup

Butyrate conversion by sulfate-reducing and methanogenic communities from anoxic sediments of Aarhus Bay, Denmark

The conventional perception that the zone of sulfate reduction and methanogenesis are separated in high-and low-sulfate-containing marine sediments has recently been changed by studies demonstrating their co-occurrence in sediments. The presence of methanogens was linked to the presence of substrates that are not used by sulfate reducers. In the current study, we hypothesized that both groups can co-exist, consuming common substrates (H2 and/or acetate) in sediments. We enriched butyrate-degrading communities in sediment slurries originating from the sulfate, sulfate–methane transition, and methane zone of Aarhus Bay, Denmark. Sulfate was added at different concentrations (0, 3, 20 mM), and the slurries were incubated at 10◦ C and 25◦ C. During butyrate conversion, sulfate reduction and methanogenesis occurred simultaneously. The syntrophic butyrate degrader Syntrophomonas was enriched both in sulfate-amended and in sulfate-free slurries, indicating the occurrence of syntrophic conversions at both conditions. Archaeal community analysis revealed a dominance of Methanomicrobiaceae. The acetoclastic Methanosaetaceae reached high relative abundance in the absence of sulfate, while presence of acetoclastic Methanosarcinaceae was independent of the sulfate concentration, temperature, and the initial zone of the sediment. This study shows that there is no vertical separation of sulfate reducers, syntrophs, and methanogens in the sediment and that they all participate in the conversion of butyrate.</p

Paleoceanographic changes in the Eastern Equatorial Pacific over the last 10 Myr

To examine the Late Neogene evolution of tropical Pacific oceanography, we determined multiple geochemical proxy records for temperature (U-37(K)' and TEX86H indices) and primary productivity (algal biomarkers and diol indices) in sediments recovered at ODP Site 1241 in the East Equatorial Pacific (EEP) spanning a record of the last 10 Myr. The TEX86H temperatures are lower than those recorded by U-37(K)' indices, exhibiting large fluctuations and suggesting strong warming during the Mid Pliocene Warm Period (MPWP; 4.5-3.2 Ma) and significantly colder temperature during the Late Miocene cooling period (7-5 Ma) and after the Middle Pliocene Warm Period (MPWP). Such variations could reflect changes in the EEP thermocline temperatures, but we suggest that they instead reflect changes in the depth of export production of glycerol dialkyl glycerol tetraether lipids in response to changes in the upper ocean structure. A combination of temperature records, inferred to represent different and likely varying depths in the water column, as well as algal biomarker records for export production and ecosystem structure, suggest that both productivity and inference upwelling were reduced in the EEP during warmer periods, such as the MPWP and prior to 7 Ma. In contrast, stronger upwelling conditions and associated increased productivity likely prevailed from 7 to 5 Ma and for the past 3 Myr, both corresponding to globally cool intervals. A further increase in EEP productivity occurred at ca 1.8 Ma, coincident with the development of the E-W Pacific SST gradient. These results confirm previous work that protracted El Nino-like conditions prevailed during warmer intervals of the Pliocene before ultimately descending into the current climate state

A unique bacteriohopanetetrol stereoisomer of marine anammox

Anaerobic ammonium oxidation (anammox) is a major process of bioavailable nitrogen removal from marine systems. Previously, a bacteriohopanetetrol (BHT) isomer, with unknown stereochemistry, eluting later than BHT using high performance liquid chromatography (HPLC), was detected in ‘Ca. Scalindua profunda’ and proposed as a biomarker for anammox in marine paleo-environments. However, the utility of this BHT isomer as an anammox biomarker is hindered by the fact that four other, non-anammox bacteria are also known to produce a late-eluting BHT stereoisomer. The stereochemistry in Acetobacter pasteurianus, Komagataeibacter xylinus and Frankia sp. was known to be 17β, 21β(H), 22R, 32R, 33R, 34R (BHT-34R). The stereochemistry of the late-eluting BHT in Methylocella palustris was unknown. To determine if marine anammox bacteria produce a unique BHT isomer, we studied the BHT distributions and stereochemistry of known BHT isomer producers and of previously unscreened marine (‘Ca. Scalindua brodeae’) and freshwater (‘Ca. Brocadia sp.’) anammox bacteria using HPLC and gas chromatographic (GC) analysis of acetylated BHTs and ultra high performance liquid chromatography (UHPLC)-high resolution mass spectrometry (HRMS) analysis of non-acetylated BHTs. The 34R stereochemistry was confirmed for the BHT isomers in Ca. Brocadia sp. and Methylocella palustris. However, ‘Ca. Scalindua sp.’ synthesise a stereochemically distinct BHT isomer, with still unconfirmed stereochemistry (BHT-x). Only GC analysis of acetylated BHT and UHPLC analysis of non-acetylated BHT distinguished between late-eluting BHT isomers. Acetylated BHT-x and BHT-34R co-elute by HPLC. As BHT-x is currently only known to be produced by ‘Ca. Scalindua spp.’, it may be a biomarker for marine anammox

Biophysical properties of membrane lipids of anammox bacteria:I. Ladderane phospholipids form highly organized fluid membranes

AbstractAnammox bacteria that are capable of anaerobically oxidizing ammonium (anammox) with nitrite to nitrogen gas produce unique membrane phospholipids that comprise hydrocarbon chains with three or five linearly condensed cyclobutane rings. To gain insight into the biophysical properties of these ‘ladderane’ lipids, we have isolated a ladderane phosphatidylcholine and a mixed ladderane phosphatidylethanolamine/phosphatidylglycerol lipid fraction and reconstituted these lipids in different membrane environments. Langmuir monolayer experiments demonstrated that the purified ladderane phospholipids form fluid films with a relatively high lipid packing density. Fluid-like behavior was also observed for ladderane lipids in bilayer systems as monitored by cryo-electron microscopy on large unilamellar vesicles (LUVs) and epi-fluorescence microscopy on giant unilamellar vesicles (GUVs). Analysis of the LUVs by fluorescence depolarization revealed a relatively high acyl chain ordering in the hydrophobic region of the ladderane phospholipids. Micropipette aspiration experiments were applied to study the mechanical properties of ladderane containing lipid bilayers and showed a relatively high apparent area compressibility modulus for ladderane containing GUVs, thereby confirming the fluid and acyl chain ordered characteristics of these lipids. The biophysical findings in this study support the previous postulation that dense membranes in anammox cells protect these microbes against the highly toxic and volatile anammox metabolites

Analysis of non-derivatized bacteriohopanepolyols using UHPLC-HRMS reveals great structural diversity in environmental lipid assemblages

Bacteriohopanepolyols (BHPs) are lipids with great chemotaxonomic potential for microbial populations and biogeochemical processes in the environment. The most commonly used methods for BHP analysis are chemical degradation followed by gas chromatography-mass spectrometry (MS) or derivatization followed by high performance liquid chromatography (HPLC)- atmospheric pressure chemical ionization/MS. Here we report on significant advances in the analysis of non-derivatized BHPs using U(ltra)HPLC-electrospray ionization-high resolution MS2. Fragmentation mass spectra provided information on the BHP core, functionalized side chain, as well as the conjugated moiety of composite BHPs. We successfully identified the common bacteriohopanepolyols and their (di)methylated and (di)unsaturated homologues, aminoBHPs, and composite BHPs (e.g., cyclitol ethers and methylcarbamateBHPs) in biomass of several known BHP-producing micro-organisms. To show how the method can be exploited to reveal the diversity of BHPs in the environment, we investigated a soil from an active methane seep, in which we detected ca. 130 individual BHPs, including a complex distribution of adenosylhopanes. We identified the nucleoside base moiety of both adenosylhopane type-2 and type-3. Adenosyl hopane type-3 contains a methylated adenine as its nucleobase, while type-2 appears to contain a deaminated and methylated adenine, or N1-methylinosine. In addition, we detected novel adenosylhopanes. Furthermore, we identified a novel series of composite BHPs comprising of bacteriohopanepolyols conjugated to an ethenolamine moiety. The novel ethanolamineBHPs as well as aminoBHPs were also detected acylated to fatty acids. The analytical approach described allows for simultaneous analysis of the full suite of IPLs, now including BHPs, and represents a further step towards environmental lipidomics

Lysine and novel hydroxylysine lipids in soil bacteria: amino acid membrane lipid response to temperature and pH in <i>Pseudopedobacter saltans</i>

Microbial decomposition of organic matter is an essential process in the global carbon cycle. The soil bacteria Pseudopedobacter saltans and Flavobacterium johnsoniae are both able to degrade complex organic molecules, but it is not fully known how their membrane structures are adapted to their environmental niche. The membrane lipids of these species were extracted and analyzed using high performance liquid chromatography-electrospray ionization/ion trap/mass spectrometry (HPLC-ESI/IT/MS) and high resolution accurate mass/mass spectrometry (HRAM/MS). Abundant unknown intact polar lipids (IPLs) from P. saltans were isolated and further characterized using amino acid analysis and two dimensional nuclear magnetic resonance (NMR) spectroscopy. Ornithine IPLs (OLs) with variable (hydroxy) fatty acid composition were observed in both bacterial species. Lysine-containing IPLs (LLs) were also detected in both species and were characterized here for the first time using HPLC-MS. Novel LLs containing hydroxy fatty acids and novel hydroxylysine lipids with variable (hydroxy) fatty acid composition were identified in P. saltans. The confirmation of OL and LL formation in F. johnsoniae and P. saltans and the presence of OlsF putative homologs in P. saltans suggest the OlsF gene coding protein is possibly involved in OL and LL biosynthesis in both species, however, potential pathways of OL and LL hydroxylation in P. saltans are still undetermined. Triplicate cultures of P. saltans were grown at three temperature/pH combinations: 30°C/pH 7, 15°C/pH 7, and 15°C/pH 9. The fractional abundance of total amino acid containing IPLs containing hydroxylated fatty acids was significantly higher at higher temperature, and the fractional abundance of lysine-containing IPLs was significantly higher at lower temperature and higher pH. These results suggest that these amino acid-containing IPLs, including the novel hydroxylysine lipids, could be involved in temperature and pH stress response of soil bacteria

Hydrogen isotopic ratios of long-chain diols reflect salinity

Long-chain diols (LCDs) are ubiquitous lipids produced by freshwater and marine algae. A combination of semi-preparative high performance liquid chromatography and gas chromatography isotope ratio monitoring mass spectrometry, allowed the measurement of δ2H of individual LCDs from cultures, which indicated a correlation with the hydrogen isotope composition of the growth water and a species-specific effect. Results from environmental samples along a salinity gradient indicated the potential of δ2H ratios of LCDs to trace the hydrogen isotopic composition of water and sea surface salinity.ISSN:0146-638

A radiocarbon-based assessment of the preservation characteristics of crenarchaeol and alkenones from continental margin sediments

Author Posting. © Elsevier B.V. , 2008. 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 Organic Geochemistry 39 (2008): 1039-1045, doi:10.1016/j.orggeochem.2008.02.006.Crenarchaeotal glycerol dibiphytanyl glycerol tetraether (GDGT) lipids and alkenones are two types of biomarkers derived from planktonic marine micro-organisms which are used for reconstruction of sea-surface temperatures. We determined the radiocarbon contents of the archaeal GDGT crenarchaeol and of alkenones isolated from continental margin sediments. Systematic differences were found between the two biomarkers, with higher radiocarbon contents in crenarchaeol than in the phytoplankton-derived alkenones. These differences can be explained by variable contributions of pre-aged, laterally advected material to the core sites. Crenarchaeol appears to be more efficiently degraded during transport in oxygen-replete environments than alkenones. Whether this reflects the influence of chemical structure or mode of protection (e.g., particle association) is not yet known.This work was funded by a Spinoza grant of NWO to J.S.S.D. and by NSF-grant OCE-0327405 to T.I.E.