Use of the pour plate technique in tuberculocidal efficacy testing according to EN 14348 - a comparative study.

Effective disinfection strategies are essential to prevent the spread of Mycobacterium tuberculosis. Tuberculocidal efficacy of disinfectants can be demonstrated by testing disinfectants in in-vitro tests, such as the well-established quantitative suspension test EN 14348 using Mycobacterium terrae as a surrogate organism in European disinfectant testing. In other European standard tests, such as EN 13727 and EN 13624, use of the pour plate technique is well established; however, in EN 14348, the spread plate technique alone is considered. Comparative experiments according to EN 14348 with M. terrae were conducted using a peracetic-acid-based disinfectant. Either the pour plate technique or the spread plate technique was used for cultivation. Differences in colony size and morphology were observed when comparing the growth of M. terrae on pour plates compared with spread plates. However, no significant differences in biocidal efficacy data were obtained when applying either spread plates or pour plates in the quantitative suspension test described in EN 14348 under both clean and dirty conditions.

Experimental fossilization of mat-forming cyanobacteria in coarse-grained siliciclastic sediments.

Microbial fossils and textures are commonly preserved in Ediacaran and early Cambrian coarse-grained siliciclastic sediments that were deposited in tidal and intertidal marine settings. In contrast, the fossilization of micro-organisms in similar marine environments of post-Cambrian age is less frequently reported. Thus, temporal discrepancies in microbial preservation may have resulted from the opening and closing of a unique taphonomic window during the terminal Proterozoic and early Phanerozoic, respectively. Here, we expand upon previous work to identify environmental factors which may have facilitated the preservation of cyanobacteria growing on siliciclastic sand, by experimentally determining the ability of microbial mats to trap small, suspended mineral grains, and precipitate minerals from ions in solution. We show that (i) fine grains coat the sheaths of filamentous cyanobacteria (e.g., Nodosilinea sp.) residing within the mat, after less than 1 week of cell growth under aerobic conditions, (ii) clay minerals do not coat sterile cellulose fibers and rarely coat unsheathed cyanobacterial cells (e.g., Nostoc sp.), (iii) stronger disturbances (where culture jars were agitated at 170 rpm; 3 mm orbital diameter) produce the smoothest and most extensive mineral veneers around cells, compared with those agitated at slower rotational speeds (150 and 0 rpm), and (iv) mineral veneers coating cyanobacterial cells are ~1 µm in width. These new findings suggest that sheathed filamentous cyanobacteria may be preferentially preserved under conditions of high fluid energy. We integrate these results into a mechanistic model that explains the preservation of microbial fossils and textures in Ediacaran sandstones and siltstones, and in fine-grained siliciclastic deposits that contain exceptionally preserved microbial mats.

Biologically agglutinated eukaryotic microfossil from Cryogenian cap carbonates.

Cryogenian cap carbonates that overlie Sturtian glacial deposits were formed during a post-glacial transgression. Here, we describe microfossils from the Kakontwe Formation of Zambia and the Taishir Formation of Mongolia-both Cryogenian age, post-Sturtian cap carbonates-and investigate processes involved in their formation and preservation. We compare microfossils from these two localities to an assemblage of well-documented microfossils previously described in the post-Sturtian Rasthof Formation of Namibia. Microfossils from both new localities have 10 ± 1 µm-thick walls composed of carbonaceous matter and aluminosilicate minerals. Those found in the Kakontwe Formation are spherical or ovoid and 90 ± 5 µm to 200 ± 5 µm wide. Structures found in the Taishir Formation are mostly spherical, 50 ± 5 µm to 140 ± 5 µm wide, with distinct features such as blunt or concave edges. Chemical and mineralogical analyses show that the walled structures and the clay fraction extracted from the surrounding sediments are composed of clay minerals, especially muscovite and illite, as well as quartz, iron and titanium oxides, and some dolomite and feldspar. At each locality, the mineralogy of the microfossil walls matched that of the clay fractions of the surrounding sediment. The abundance of these minerals in the walled microfossils relative to the surrounding carbonate matrix and microbial laminae, and the presence of minerals that cannot precipitate from solution (titanium oxide and feldspar), suggests that the composition represents the original mineralogy of the structures. Furthermore, the consistency in mineralogy of both microfossils and sediments across the three basins, and the uniformity of size and shape among mineral grains in the fossil walls indicate that these organisms incorporated these minerals by primary biological agglutination. The discovery of new, mineral-rich microfossil assemblages in microbially laminated and other fine-grained facies of Cryogenian cap carbonates from multiple localities on different palaeocontinents demonstrates that agglutinating eukaryotes were widespread in carbonate-dominated marine environments in the aftermath of the Sturtian glaciation.

Molecular biosignatures reveal common benthic microbial sources of organic matter in ooids and grapestones from Pigeon Cay, The Bahamas.

Ooids are sedimentary grains that are distributed widely in the geologic record. Their formation is still actively debated, which limits our understanding of the significance and meaning of these grains in Earth's history. Central questions include the role played by microbes in the formation of ooids and the sources of ubiquitous organic matter within ooid cortices. To address these issues, we investigated the microbial community composition and associated lipids in modern oolitic sands at Pigeon Cay on Cat Island, The Bahamas. Surface samples were taken along a transect from the shallow, turbulent surf zone to calmer, deeper water. Grains transitioned from shiny and abraded ooids in the surf zone, to biofilm-coated ooids at about 3 m water depth. Further offshore, grapestones (cemented aggregates of ooids) dominated. Benthic diatoms and Proteobacteria dominated biofilms. Taxa that may promote carbonate precipitation were abundant, particularly those associated with sulfur cycling. Compared to the lipids associated with surface biofilms, relict lipids bound within carbonate exhibited remarkably similar profiles in all grain types. The enhanced abundance of methyl-branched fatty acids and ß-hydroxy fatty acids, 1-O-monoalkyl glycerol ethers and hopanoids bound within ooid and grapestone carbonate confirms a clear association of benthic sedimentary bacteria with these grains. Lipids bound within ooid cortices also contain molecular indicators of microbial heterotrophic degradation of organic matter, possibly in locally reducing conditions. These included the loss of labile unsaturated fatty acids, enhanced long-chain fatty acids/short-chain fatty acids, enriched stable carbon isotopes ratios of fatty acids, and very high stanol/stenol ratios. To what extent some of these molecular signals are derived from later heterotrophic endolithic activity remains to be fully resolved. We speculate that some ooid carbonate forms in microbial biofilms and that early diagenetic degradation of biofilms may also play a role in early stage carbonate precipitation around ooids.

Lipid biomarkers in ooids from different locations and ages: evidence for a common bacterial flora.

Ooids are one of the common constituents of ancient carbonate rocks, yet the role that microbial communities may or may not play in their formation remains unresolved. To search for evidence of microbial activity in modern and Holocene ooids, samples collected from intertidal waters, beaches and outcrops in the Bahamas and in Shark Bay in Western Australia were examined for their contents of lipid biomarkers. Modern samples from Cat and Andros islands in the Bahamas and from Carbla Beach in Hamelin Pool, Western Australia, showed abundant and notably similar distributions of hydrocarbons, fatty acids (FAs) and alcohols. A large fraction of these lipids were bound into the carbonate matrix and only released on acid dissolution, which suggests that these lipids were being incorporated continuously during ooid growth. The distributions of hydrocarbons, and their disparate carbon isotopic signatures, were consistent with mixed input from cyanobacteria together with small and variable amounts of vascular plant leaf wax [C27 -C35 ; δ(13) C -25 to -32‰Vienna Pee Dee Belemnite (VPDB)]. The FAs comprised a complex mixture of C12 -C18 normal and branched short-chain compounds with the predominant straight-chain components attributable to bacteria and/or cyanobacteria. Branched FA, especially 10-MeC16 and 10-MeC17 , together with the prevalence of elemental sulfur in the extracts, indicate an origin from sulfate-reducing bacteria. The iso- and anteiso-FA were quite variable in their (13) C contents suggesting that they come from organisms with diverse physiologies. Hydrogen isotopic compositions provide further insight into this issue. FAs in each sample show disparate δD values consistent with inputs from autotrophs and heterotrophs. The most enigmatic lipid assemblage is an homologous series of long-chain (C24 -C32 ) FA with pronounced even carbon number preference. Typically, such long-chain FA are thought to come from land plant leaf wax, but in this case, their (13) C-enriched isotopic signatures compared to co-occurring n-alkanes (e.g., Hamelin Pool TLE FA C24 -C32 ; δ(13) C -20 to -24.2‰ VPDB; TLE n-alkanes δ(13) C -24.1 to -26.2 -‰VPDB) indicate a microbial origin, possibly sulfate-reducing bacteria. Lastly, we identified homohopanoic acid and bishomohopanol as the primary degradation products of bacterial hopanoids. The distributions of lipids isolated from Holocene oolites from the Rice Bay Formation of Cat Island, Bahamas were very similar to the beach ooids described above and, in total, these modern and fossil biomarker data lead us to hypothesize that ooids are colonized by a defined microbial community and that these microbes possibly mediate calcification.

Molecular indicators of microbial diversity in oolitic sands of Highborne Cay, Bahamas.

Microbialites (stromatolites and thrombolites) are mineralized mat structures formed via the complex interactions of diverse microbial-mat communities. At Highborne Cay, in the Bahamas, the carbonate component of these features is mostly comprised of ooids. These are small, spherical to ellipsoidal grains characterized by concentric layers of calcium carbonate and organic matter and these sand-sized particles are incorporated with the aid of extra-cellular polymeric substances (EPS), into the matrix of laminated stromatolites and clotted thrombolite mats. Here, we present a comparison of the bacterial diversity within oolitic sand samples and bacterial diversity previously reported in thrombolitic and stromatolitic mats of Highborne Cay based on analysis of clone libraries of small subunit ribosomal RNA gene fragments and lipid biomarkers. The 16S-rRNA data indicate that the overall bacterial diversity within ooids is comparable to that found within thrombolites and stromatolites of Highborne Cay, and this significant overlap in taxonomic groups suggests that ooid sands may be a source for much of the bacterial diversity found in the local microbialites. Cyanobacteria were the most diverse taxonomic group detected, followed by Alphaproteobacteria, Gammaproteobacteria, Planctomyces, Deltaproteobacteria, and several other groups also found in mat structures. The distributions of intact polar lipids, the fatty acids derived from them, and bacteriohopanepolyols provide broad general support for the bacterial diversity identified through analysis of nucleic acid clone libraries.