Bacterial trace fossils in eocene kaolin of the huber formation of Georgia; phylloderma microsphaeroides,n. ichnogen., n. ichnosp.

An Eocene kaolin deposit in Georgia contains microspheres of tangentially oriented kaolinite enclosing authigenic cristobalite/tridymite overgrown with kaolinite. These structures resemble cutans of clay minerals that surround polysaccharide‐covered bacteria in recent soils and sediments. Bacteria actively synthesize anionic mucopolysaccharides as adhesives for attachment to surfaces. Empty cutans are common; these polymer‐bound clay mineral microspheres persist following cell death and lysis. Even though the void space thus created and associated organic material influence subsequent diagenetic changes, sedimentary microfabrics attributable to bacterial activity are preserved.     

Effects of Different Clay’s Percentages on Improvement of Sand-Clay Mixtures with Microbially Induced Calcite Precipitation

Abstract

Microbially induced calcite precipitation (MICP) is currently appraised for the improvement of sandy soils, but only few studies use it to improve sand-clay mixtures. The effect of contents of kaolin clay and the effect of ions in kaolin clay on bacterial urease activity and productive rates for calcium carbonate were studied. Moreover, sand solidification tests were conducted and the solidifying effects of MICP for sand-clay mixtures were evaluated. The results show that adding kaolin clay has an inhibitory effect on the urease activity of bacteria, and adding too many kaolin clays also decrease the productive rates for calcium carbonate. With adding Al₂O₃ or FeCl₃, urease activity both decreases and it becomes lower with adding more Al₂O₃ or FeCl₃. The permeability of sand columns all decreased gradually with MICP curing. With more kaolin clay, the increasing range of bacterial utilization rates of those with larger particle sizes is bigger. The maximum productive rate for calcium carbonate of samples with smaller particle sizes exists in sample with 5% of kaolin clay while other samples with 7.5% of clay have more calcium carbonate. Sand columns with different sand particle sizes have different suitable amounts of added kaolin clays for MICP solidification.     

Adsorption of lecithin liposomes to acid clay

The interaction between lecithin liposomes and acid clay was investigated to clarify the mechanism for liposome adsorption to the clay. It was found that the multilamellar vesicular structure of the liposomes was broken as a result of primary adsorption. The acid clay particles aggregated and were eventually covered by the lecithin layer structure. In the case of kaolin, on the other hand, the liposomes were weakly adsorbed to the clay and maintained the vesicular structure. The amount of primary adsorption to the clay surface, which was estimated from the adsorption isotherm, was more for acid clay than for kaolin, and the total amount adsorbed to the acid clay was also more than to kaolin. This result can be explained by the much higher density of the negative charge on the acid clay surface than that for kaolin. The liposomes are therefore considered to be adsorbed to the acid clay mainly by the choline positive charge residing at the end of the lecithin molecule, although this is of no net charge as a whole.     

The radiolysis and radioracemization of amino acids on clays

L-Leucine and its hydrochloride salt have been deposited on the clay minerals kaolin and bentonite, and the amino acid/clay preparations have been irradiated in a 3000 Ci60Co gamma-ray source for radiation dosages that achieved 2-89% radiolysis of the leucine. The undecomposed leucine was thereupon recovered and both percent radiolysis and percent radioracemization were determined. Similar studies were made using solid L-leucine and its hydrochloride, and L-leucine in 0.1 M aqueous solution. It has been found that radiolysis and radio-racemization in these and the previously studied leucine systems follow pseudo-first-order rate laws, and the corresponding specific rate constants are evaluated and compared. Leucine and its hydrochloride salt proved to be the most stable to both radiolysis and radioracemization, followed by leucine and its HCl salt on kaolin, followed by leucine and its HCl salt on bentonite, with leucine (and its HCl and Na salts) in aqueous solution being least stable to both radiolysis and (except for the HCl salt) radioracemization. Implications of these observations as regards the Vester-Ulbricht mechanism for the origin of optical activity are discussed.     

Effects of kaolin on the composition of generalist predator assemblages and parasitism of Choristoneura rosaceana (Lep., Tortricidae) in apple orchards

Abstract:  In three apple orchards, we tested how the hydrophilic kaolin clay particle film Surround WP affected the diversity of generalist arthropod predator assemblages in orchard foliage and the parasitism of the pest species Choristoneura rosaceana (Harris) (obliquebanded leafroller) (Lep., Tortricidae). In two orchards, kaolin was applied to orchard foliage once a week for 4 weeks, between mid-June and mid-July in 2004 and 2005. In the third orchard kaolin was applied to foliage twice over 2 weeks in June 2004. We quantified the proportion of larvae C. rosaceana parasitized, larval populations, and the relative abundance and assemblage composition of generalist predators (spiders and insects) in the orchards. Kaolin altered the species composition of the generalist predator assemblages and reduced the relative abundances of certain generalist predators, most notably jumping and crab spiders (Salticidae and Philodromidae), assassin bugs (Reduviidae), ants (Formicidae) and coccinellids (Coccinellidae). In contrast, the relative abundances of web-spinning spiders (Araneidae, Dictynidae, Theridiidae) were not affected. Kaolin did not affect the proportion of parasitized C. rosaceana larvae, which ranged from 24% to 47% in control and kaolin treatments, or the relative proportions of parasitoid taxa. The kaolin formulation did not affect the abundance of C. rosaceana larvae, but in one orchard, kaolin did reduce the abundance of the combined numbers of C. rosaceana and another tortricid pest, Argyrotaenia velutiana . Although kaolin does not affect parasitism of C. rosaceana , it significantly changes the composition of generalist predator assemblages in orchard foliage.     

Drinking rate regulation in some fresh-water mosquito larvae

ABSTRACT. Drinking rates were determined with four species of freshwater mosquito larvae by colorimetric measurement of the dye ingested after groups of fourth instars were allowed access for set periods to 2% amaranth solutions. The rate of drinking for the saline-tolerant Aedes aegypti, 309±113 nl per h per individual, was comparable with rates given in the literature for several saline-water species, but rates for Culex quinquefasciatus, Culex molestus and Anopheles albimanus were markedly lower (167±30, 48±17 and 108±28m per h, respectively). When larvae of A.aegypti and C. quinquefasciatus were glutted with kaolin (allowed to replace all food in the gut by filter-feeding in kaolin suspension), drinking rates were little affected at first, but after 1 day of fasting (holding in water after glutting), drinking rates were 50% lower and were reduced by a further 20% with fasting for up to 3 days. For A.aegypti, C.quinquefasciatus and C. molestus, drinking rates were approximately doubled with kaolin dispersed in the dye solution, and after fasting, were increased by up to 100% in solutions containing 0.05% of water-soluble yeast extract. A similar phagostimulant effect of 10^-3M adenylic acid was demonstrated for A. aegypti and C. quinquefasciatus. A single experiment indicated similar stimulatory effects of kaolin and adenylic acid for, A. albimanus. With 0.01-0.05% agarose in the dye solutions, drinking rates for A. aegypti and C. quinquefasciatus were more than doubled, and a similar though weaker effect was demonstrated for another colloid, methylcellulose. In constrast, both colloids markedly reduced the rate of drinking with A.albimanus. These findings are discussed in relation to whether drinking and filter-feeding are necessarily coupled. The possible significance of this with respect to larvae that feed in different microhabitats, providing different levels of dissolved and colloidal nutrient organic matter, is considered. The implications of drinking rates for biotests of solubilized bacterial toxins as mosquito larvicides are noted.     

Effects of kaolin on Ophelimus maskelli (Hymenoptera: Eulophidae) in laboratory and nursery experiments

Although recent research has demonstrated that clays provide satisfactory control of some agricultural insect pests, the effect of clays on gall wasps that damage forest trees has not been previously reported. The aim of the current study is to evaluate the effectiveness of the clay kaolin in the laboratory and in the field in reducing the damage caused by the eulophid Ophelimus maskelli (Ashmead) on seedlings of eucalyptus (Eucalyptus L'Hér.) species. In the laboratory, kaolin + wetting agent significantly reduced the percentage of infested leaves and the number of galls per leaf. In the nursery, gall number per leaf was not correlated with leaf area with kaolin + wetting agent but was related to leaf area for all other treatments (wetting agent alone, imidacloprid, and untreated control). In the nursery, gall number per leaf was lower with kaolin + wetting agent and with imidacloprid than with the other two treatments. Overall, kaolin effectively reduced eulophid infestations, and its effect was more persistent than that of imidacloprid. Although application of kaolin might not be feasible on large forested areas, kaolin could represent a valuable control method in nurseries, where the repeated application with more toxic chemicals can result in high concentrations of residual pesticides in the soil.     

Application of living immobilized cells to the acceleration of the continuous conversions of ethanol (wort) to acetic acid (vinegar)—Hydrous titanium(IV) oxide-immobilized Acetobacter species

Various strains of Acetobacter species have been immobilized on hydrous titanium(IV) oxide or hydrous titanium(IV) chelated cellulose and used in the continuous conversion of a dilute aqueous alcoholic solution (in the form of‘charging wort’) into acetic acid (in the form of vinegar) in tower fermenter-type reactors. A strain of Acetobacter species producing extracellular polysaccharide aggregated in the presence of hydrous titanium(IV) oxide thereby enabling higher medium flow rates and an increased acetic acid output to be achieved. A strain of Acetobacter species not producing polysaccharide showed no effect with hydrous titanium(IV) oxide but did produce more acetic acid when a titanium(IV)-cellulose chelate was added to the fermentation, although aggregation was not observed. Mechanisms, which appear to conform to established results, are proposed for the aggregation of both strains of bacteria. Apparently, these water-insoluble titanium compounds can interact with the bacterial cells, increasing their density and thus making them more resistant to ‘wash out’ by increasing the rate at which they sediment in the fermenter. This enables a greater cell mass per unit volume to be achieved which in turn leads to an increase in conversion rate in the reactor.     

Isolation and characteristics of a specific polysaccharide of group-B meningococci

The optimum conditions for the isolation and purification of the specific polysaccharide of group B meningococci have been developed. The advantages of the use of synthetic culture media for growing the initial bacterial culture have been demonstrated. The purified polysaccharides have been found to contain about 70% of sialic acid and less than 1% of protein and nucleic acid admixtures. The molecular parameters of group B polysaccharide depended on the growth phase of the bacterial culture. The most valuable culture was obtained at the exponential phase of growth. High serological activity and specificity of the polysaccharide in the passive hemagglutination test recommend it for studies on the development of diagnostic and prophylactic preparations.     

The extracellular polysaccharides of Rhizobium japonicum: Compositional studies

The extracellular polysaccharides of seven strains of Rhizobium japonicum were investigated by using a gas-chromatographic scheme developed for determination of the various sugars present. These polysaccharides were more heterogeneous in their composition than those of any other species of Rhizobium yet examined. Five strains (1809, 110, 123, 127, and 709) produced polysaccharides containing the same constituents, although in varying relative amounts: glucose (36–44%), galactose (7–25%), mannose (18–20%), 4-O-methylgalactose (5–13%), galacturonic acid (12–16%), and acetyl groups (4–8%). The sugars of the polysaccharide of strain 1809 were all of the d series. These are the first bacterial polysaccharides reported to contain 4-O-methylgalactose and the first Rhizobium polysaccharides in which galacturonic acid has been found. In contrast to this, the polysaccharide of strain 129 consisted of glucose (7%), galactose (51%), mannose (5%), xylose (5%), glucuronic acid (5%), and pyruvic acid (2%). The polysaccharide of strain 711 contained glucose (34%), galactose (13%), mannose (27%), and pyruvic acid (6%).     

Disaccharide analysis and molecular mass determination to microgram level of single sulfated glycosaminoglycan species in mixtures following agarose-gel electrophoresis.

The separation of sulfated glycosaminoglycans in mixtures by agarose-gel electrophoresis and the recovery of single polysaccharide bands has been applied to the characterization of polysaccharides extracted from tissues without previous purification of single species. Sulfated glycosaminoglycans, heparin with its two components, slow-moving and fast-moving, heparan sulfate, dermatan sulfate, and chondroitin sulfate, were separated to microgram level by conventional agarose-gel electrophoresis. After their separation, they were fixed in the agarose-gel matrix by precipitation in a cetyltrimethylammonium bromide solution, making them visible on a dark background. After recovery of gel containing the fixed bands, high temperatures (90 degrees C for 15 min) were necessary to dissolve the gel matrix, and a solution of NaCl (3 M) was used to release sulfated polysaccharides from the complex with cetyltrimethylammonium. After precipitation of glycosaminoglycans in the presence of ethanol, the recovery of slow-moving heparin, fast-moving heparin, heparan sulfate, dermatan sulfate, and chondroitin sulfate was from 1 to 10 microg, with a percentage greater than 45% and a purity above 90%. Sulfated glycosaminoglycans in mixtures recovered from gel matrix as single species were evaluated for purity and characterized for unsaturated disaccharides after treatment with bacterial lyases (heparinases for heparin and heparan sulfate samples, and chondroitinases for dermatan sulfate and chondroitin sulfate) and molecular mass. Bovine lung and heart Glycosaminoglycans were extracted and separated into single species by agarose-gel electrophoresis and recovered from gel matrix after treatment in cetyltrimethylammonium solution. Unsaturated disaccharides pattern, the sulfate to carboxyl ratio, and the molecular mass of each single polysaccharide species were determined.     

POLYSACCHARIDE RELEASE BY APHANOTHECE HALOPHYTICA INHIBITS CYANOBACTERIA/CLAY FLOCCULATION1

Many microalgae release polysaccharides, but the effects of the polysaccharides on mutual flocculation of microalgae and clay in aquatic environments have not been well studied. Aphanothece halophytica Frémy is a bloom‐forming cyanobacterium in salterns and can release large amounts of polysaccharide (AH‐RPS). In the present study, we investigated the effect of AH‐RPS on mutual flocculation of cyanobacterium and clay and further explored the mechanisms by which AH‐RPS affected mutual flocculation. We determined that AH‐RPS possessed clay‐dispersing activity as defined as the ability to inhibit the flocculation and sedimentation of clay suspensions in water. Supplementation of AH‐RPS in cyanobacterial cell suspensions and in the culture media containing the same kaolin clay concentration dose dependently decreased flocculation of cyanobacterial cells and increased clay‐dispersing activity. These findings indicate that the clay‐dispersing activity of AH‐RPS was related to its inhibitory effect on mutual flocculation of cyanobacterial cells and clay particles. Moreover, the clay‐dispersing activity of AH‐RPS was stable from pH 3 to pH 10 and was increased by adding NaCl, MgCl₂, CaCl₂, or low concentrations of KCl (up to 0.4 M). Taken together, our data suggest that AH‐RPS might maintain its clay‐dispersing activity and inhibit mutual flocculation of microalgae and suspended clay in saltern brine.     

Anaerobic decolorisation of simulated textile wastewater containing azo dyes

The effects of the addition of powered particles of kaolin to nitrifying activated sludge systems were studied. Kaolin was added to a nitrifying activated sludge reactor, during the operational phase, to observe the effects of this clay on reactor performance. The results were compared to those obtained from a similar unit operated without kaolin. The settling properties of the sludges from both units were similar (sludge volume index (SVI) of 14.5 ml/g VSS; zone settling velocity (ZSV) of 7.5 m/h), but the specific nitrifying activities of ammonia and nitrite oxidizing processes were enhanced up to 75% and 50%, respectively, when kaolin was added. The mechanism of action of kaolin was not clear. Additional ammonia, nitrite and nitrate adsorption tests showed that these compounds were not adsorbed by kaolin. This demonstrated that no beneficial effect was caused by adsorption of either substrates or products. Short-term activity tests also showed that the stimulating effects of kaolin on specific activity were not immediate. The effects of kaolin when nitrifying units were operated under unfavorable conditions were also evaluated: In a second set of experiments, a nitrifying unit was operated with low levels of dissolved oxygen (DO), with and without kaolin. The presence of kaolin exerted practically no effect on ammonia oxidation but nitrite oxidation slightly diminished. In a third set of experiments, a nitrifying unit was subjected to pH shocks (9, 10 and 11) over 3 h with pH then restored to 7.8. A pH shock of 11 caused a decrease of 60% in nitrifying activity for 12 days. When kaolin was added to this unit the efficiency of the system was completely restored in 4 days. Therefore, kaolin might be useful to restore damaged units.     

A lipopolysaccharide fromAspergillus flavus conidia

A lipopolysaccharide was isolated by extraction ofAspergillus flavus conidia with 45 % phenol at 68–70 °C. Quantitative analysis revealed 7 % nucleic acids, 5.5 % proteins, 46 % polysaccharides and 49 % lipids, of which 12 % were covalently bound. Glucose, mannose, galactose and fucose were detected as monosaccharide components of the polysaccharide moiety by gas chromatography; palmitic acid, stearic acid, oleic acid, linoleic acid and myristic acid were mainly present in the lipidic fraction. This material differs from the bacterial lipopolysaccharides, both in composition of the polysaccharide moiety and representation of fatty acids in the lipidic fraction.     

Quantitative determination of pneumococcal capsular polysaccharide serotype 14 using a modification of phenol-sulfuric acid method

The capsular polysaccharide of Streptococcus pneumoniae, serotype 14, is part of every pneumococcal vaccine presently in the market or under development. A strategy for the quantitative determination of this polysaccharide by the phenol-sulfuric acid method is described. The modality of acid addition is shown to be the critical step for obtaining reproducible test results between different technicians. Raising the incubation temperature above 80 degrees C increased the consistency of the method by more than 60% regardless of the acid addition modality, but at the expense of some loss of sensitivity. Incubation at 110 degrees C was found necessary to obtain reproducible results within 3% for this technique, which was used to follow the enrichment of the polysaccharide during the last steps of purification. A model mixture of the component polysaccharide sugars provided an adequate and economic standard to construct the calibration curve for this assay, with absorbance reading either in the reaction tubes or in a microplate. A similar procedure may be applied to the determination of other bacterial polysaccharides as well.     

On the asymmetric polymerization of aspartic acid enantiomers by kaolin

Summary Two recent reports in the literature claim thatl-aspartic acid polymerizes significantly faster thand-aspartic acid in the presence of kaolin in aqueous solution at 90°. The novelty of these observations and their potential significance for molecular evolution and the origin of optical activity in nature has prompted us to attempt a duplication of the experiments involved—using, however, analytical criteria which we felt would be more reliable than those previously employed. In our experimentsl- andd,l-aspartic acid in 0.01M solution were incubated with kaolin at 90° for 8 days. Careful examination of the aqueous residues from such experiments, however, failed to demonstrate any preferential polymerization ofl- overd-aspartic acid under the influence of kaolin, or indeed any significant gross polymerization of aspartic acid at all.     

Bacterial community structure and activity in different Cd-treated forest soils

In this study we compared indicators of Cd bioavailability (water extracts, Lakanen extracts, free ions) and ecotoxicity in forest soils with contrasting physico-chemical characteristics. Soil samples were treated with CdCl(2) solutions (0, 0.1, 1, 10 and 100 mM) and incubated for 30 days. Microbial activity indexes (acid phosphatase, beta-glucosidase, basal respiration) and changes in bacterial community structure using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting were investigated. The Cd concentrations measured ranged from 1% to 37% of the total additions in water extracts, to higher levels in Lakanen extracts. Effects of Cd were observed at bioavailable concentrations exceeding United Nations/European Economic Commission UN/ECE guidelines for total Cd in the soil solution. Basal respiration was the most affected index, while enzymatic activities showed variable responses to the Cd treatments. We also noticed that soils with pH higher than 6.7 and clay content higher than 50% showed inhibition of basal respiration but no marked shift in bacterial community structure. Soils with lower pH (pH <5.8) with less clay content (<50%) showed in addition strong changes in the bacterial community structure. Our results provide evidence for the importance of relating the effects of Cd on the soil communities to soil properties and to bioavailability.     

Conformational and dynamic differences between N. meningitidis serogroup B and C polysaccharides, using n.m.r. spectroscopy and molecular mechanics calculations

1H-N.m.r. spectroscopy has been used to determine the conformation in aqueous solution of the sialic acid residues of the N. meningitidis serogroup B and non-O-acetylated (O-Ac-)-C polysaccharides, and of N-acetylneuraminic acid (NeuNAc). In all cases, the sugar adopts the 2C5 conformation. The side-chain of NeuNAc adopts a conformation such that H-7 and H-8 are approximately anti-periplanar. This conformation is also found in the (O-Ac-)-C polysaccharide, whereas H-7 and H-8 are gauche in the B polysaccharide. Molecular mechanics calculations have been used to probe the conformational preferences of the variously linked sialic acid residues, and the results are in general agreement with those based on the 1H-n.m.r. data. The 13C-n.m.r. spin-lattice relaxation-times have been interpreted in terms of the molecular dynamics of the B and (O-Ac-)-C polysaccharides. Molecular correlation times have been calculated and details of internal rotational or segmental motion elucidated. The C polysaccharide is characterised by internal or segmental motion in the C-7 to C-9 side-chain of the sialic acid repeating-unit, whereas the B polysaccharide has little or no such movement and tumbles in solution as a rigid species with internal rotation of only the pendant C-9 group. The conformational differences suggest a substantially different three-dimensional structure in solution for these polysaccharides.     

Formation of Novel Polysaccharides by Bradyrhizobium japonicum Bacteroids in Soybean Nodules

Certain strains of Bradyrhizobium japonicum form a previously unknown polysaccharide in the root nodules of soybean plants (Glycine max (L.) Merr.). The polysaccharide accumulates inside of the symbiosome membrane—the plant-derived membrane enclosing the bacteroids. In older nodules (60 days after planting), the polysaccharide occupies most of the symbiosome volume and symbiosomes become enlarged so that there is little host cytoplasm in infected cells. The two different groups of B. japonicum which produce different types of polysaccharide in culture produce polysaccharides of similar composition in nodules. Polysaccharide formed by group I strains (e.g., USDA 5 and USDA 123) is composed of rhamnose, galactose, and 2-O-methylglucuronic acid, while polysaccharide formed by group II strains (e.g., USDA 31 and USDA 39) is composed of rhamnose and 4-O-methylglucuronic acid. That the polysaccharide is a bacterial product is indicated by its composition plus the fact that polysaccharide formation is independent of host genotype but is dependent on the bacterial genotype. Polysaccharide formation in nodules is common among strains in serogroups 123, 127, 129, and 31, with 27 of 39 strains (69%) testing positive. Polysaccharide formation in nodules is uncommon among other B. japonicum serogroups, with only 1 strain in 18 (6%) testing positive.     

Characterization of cpsF and its product CMP-N-acetylneuraminic acid synthetase, a group B streptococcal enzyme that can function in K1 capsular polysaccharide biosynthesis in Escherichia coli

Group B Streptococcus (GBS) is the foremost cause of neonatal sepsis and meningitis in the United States. A major virulence factor for GBS is its capsular polysaccharide, a high molecular weight polymer of branched oligosaccharide subunits. N -acetylneuraminic acid (Neu5Ac or sialic acid), at the end of the polysaccharide side chains, is critical to the virulence function of the capsular polysaccharide. Neu5Ac must be activated by CMP-Neu5Ac synthetase before it is incorporated into the polymer. We showed previously that a transposon mutant of a serotype III GBS strain which had no detectable capsular Neu5Ac was deficient in CMP-Neu5Ac-synthetase activity (Wessels et al ., 1992). In this paper, we report the identification and characterization of cpsF , a gene interrupted by transposon insertion in the previously described Neu5Ac-deficient mutant. The predicted amino acid sequence of the cpsF gene product shares 57% similarity and 37% identity with CMP-Neu5Ac synthetase encoded by the Escherichia coli K1 gene, neuA . The enzymatic function of the protein encoded by cpsF was established by cloning the gene in E. coli under the control of the T7 polymerase/promoter. Lysates of E. coli in which the cpsF gene product was expressed, catalysed the condensation of CTP with Neu5Ac to form CMP-Neu5Ac. In addition, when a CMP-Neu5Ac synthetase-deficient mutant of E. coli K1 was transformed with cpsF , K1 antigen expression was restored. We conclude that cpsF encodes CMP-Neu5Ac synthetase in type III GBS, and that the GBS enzyme can function in the capsule-synthesis of a heterologous bacterial species.