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Biomedical subjects

Nicholas J Russell

Publications and source records attributed to Nicholas J Russell.

10 recordsLinked to original sources

The influence of different temperature programmes on the bioremediation of polycyclic aromatic hydrocarbons (PAHs) in a coal-tar contaminated soil by in-vessel composting.

The biodegradation of 16 US. EPA-listed polycyclic aromatic hydrocarbons (sigma PAHs), with accompanying humification and microbial community structure changes during simulated in-vessel composting-bioremediation of an aged coal-tar-contaminated soil amended with green waste were studied over 56 days. The experimental design compared one constant temperature profile (TC=38 degrees C) with three variable temperature profiles (TP1, TP2 and TP3), including treatment at 70 degrees C to comply with regulatory requirements. Greatest sigma PAHs removal (75.4+/-0.1%; k(1)=0.026 day(-1), R(2)=0.98) occurred at TC=38 degrees C compared to all variable temperature profiles TP1 (62.1+/-11.0%; k(1)=0.016 day(-1), R(2)=0.93), TP2 (71.8+/-8.2%; k(1)=0.021 day(-1), R(2)=0.95) and TP3 (45.3+/-9.7%; k(1)=0.010 day(-1), R(2)=0.91). This study proved that using thermophilic temperatures (70 degrees C) towards the end of in-vessel composting processes (TP2) resulted in greater sigma PAHs removal than using other variable temperature profiles (TP1, TP3), as long as the increase was stepwise via an intermediate temperature (55 degrees C). Phospholipid fatty acid (PLFA) signatures indicated that use of thermophilic temperatures towards the end of the in-vessel composting-bioremediation (TP2) resulted in a higher fungal to bacterial PLFA ratio and a lower Gram-positive to Gram-negative (G(+)/G(-)) bacterial ratio. Fluorescence excitation-emission matrix (EEM) showed the presence of peaks typical of humic-like (Ex/Em wavelength pair approximately 340/460 nm) and fulvic-acid-like (Ex/Em wavelength pair approximately 245/460 nm) substances, indicating mineralization and/or maturation of the compost. Varying the temperature during in-vessel composting to comply with regulatory requirements for pathogen control, promoted contaminant biodegradation, microbial activity and compost maturation.

Bacteria↗

Phenotypic and genotypic characterisation of Bacillus cereus isolates from Bangladeshi rice.

We report the first phenotypic and genotypic characterisation of Bacillus cereus from Bangladeshi rice. Seven strains of B. cereus were isolated from two high-yielding rice varieties (BR5 and BRRI Dhan28) which are cultivated during different rice-growing seasons in Bangladesh. The strains were identified as B. cereus based on colonial and cellular (light microscopic) morphology and were confirmed using the API 50 CHB test system and 16S rDNA analysis. They could also be distinguished from each other based on their substrate-utilisation patterns using the API system. No psychrotrophic (cold-tolerant) isolates were obtained and all were mesophilic with a minimum growth temperature of 10 degrees C, but with an ability to grow at a higher rate than normal temperature (45 degrees C) for B. cereus. All strains had temperature optima between 35 and 45 degrees C, but two groups could be distinguished based on having either a narrow or a broad optimum range. The different strains could not be distinguished based on their phospholipid compositions including major fatty acyl chains which were typical for B. cereus. However, capillary-GC/MS analysis of trace fatty acyl components of the lipids has been used for the first time to group the isolates. Genotypic analysis of the rDNA region has also been used to distinguish the strains. Restriction fragment length polymorphism (RFLP) analysis of the spacer region between the 16S and 23S rRNA genes yielded two groups based on different base sequences, whereas sequence analysis of variable regions in the 16S rRNA gene gave four different groups by base differences at two positions. Despite their association with rice, none of the strains produced emetic toxin. The spore germination response of heat-activated spores of the Bangladeshi B. cereus strains was typical of the bacterium, but three different groups could be identified based on differences in germination rates. The use of this polyphasic approach to taxonomy has enabled all the B. cereus strains to be distinguished from each other.

Bacillus cereus↗

Strains of Bacillus cereus vary in the phenotypic adaptation of their membrane lipid composition in response to low water activity, reduced temperature and growth in rice starch.

The phenotypic adaptation of membrane lipids in seven strains of the food-poisoning bacterium Bacillus cereus, isolated from Bangladeshi rice, is reported in relation to their ability to grow under conditions of low water activity (a(w)), reduced temperature and the presence of soluble rice starch. The strains have different membrane phospholipid head-group and fatty acyl compositions, and they display individual differences in their responses to both low a(w) and reduced temperature. The extent of the increase in anionic membrane lipids in response to low a(w) varies from strain to strain, is solute specific and in one strain does not occur. Growth is stimulated by the presence of soluble rice starch and results in a large rise in the proportion of diphosphatidylglycerol (DPG) at the expense of phosphatidylglycerol (PG), without any change in the proportion of total anionic phospholipids. Growth at 15 degrees C compared with 37 degrees C increases the proportions of DPG and phosphatidylethanolamine at the expense of PG. At the lower temperature there are changes in phospholipid fatty acyl composition characteristic of those expected to maintain membrane fluidity, including increases in the amount of total branched fatty acids and the anteiso-/iso-branched ratio, and a decrease in the equivalent chain-length, but there are strain differences in how those changes were achieved. In contrast to some other bacilli, there are persistent large increases in the proportions of unsaturated fatty acyl chains in phospholipids during growth at 15 degrees C.

Adaptation, Physiological↗

Rhodopseudomonas acidophila strain 10050 contains photosynthetic LH2 antenna complexes that are not enriched with phosphatidylglycerol, and the phospholipids have a fatty acyl composition that is unusual for purple non-sulfur bacteria.

The phospholipid composition of Rhodopseudomonas acidophila strain 10050 grown aerobically or anaerobically in the light was determined. The major phospholipids present in the aerobic cells were phosphatidylethanolamine (PE; 54%), phosphatidylglycerol (PG; 24%) and cardiolipin (diphosphatidylglycerol, DPG) (14%), together with phosphatidylcholine (PC; 5%). On moving the cells to anaerobic photosynthetic growth in the light PE remained the major phospholipid (37-49%), but there was a major change in the proportion of PC, which increased to 31-33%, and corresponding reductions in the contents of PG to 11-16% and DPG to 4-5%. The fatty acid composition of the phospholipids was unusual, compared with other purple non-sulfur photosynthetic bacteria, in that it contained 16:0 (29%), 17:1 (20%) and 19:1 (9%) plus several mainly unsaturated 2-OH fatty acids (9% total) as major components, when grown aerobically in the dark. In contrast when grown photosynthetically under anaerobic conditions there was <2% 17:1 or 19:1 present, while the amounts of 16:1 and 18:1 increased, and 16:0 decreased. The phospholipid composition of the purified light-harvesting complex 2 (LH2) complex was PE (43%), PC (42%) and DPG (15%). Unexpectedly, there was no PG associated with the purified LH2. These findings contrast with previous studies on several other photosynthetic bacteria, which had shown an increase in PG upon photosynthetic growth [Biochem. J. 181 (1979) 339]. The prior hypothesis that phosphatidylglycerol has some specific role to play in the function of light-harvesting complexes cannot be true for Rps. acidophila. It is suggested that specific integral membrane proteins may strongly influence the phospholipid content of the host membranes into which they are inserted.

Bacterial Proteins↗

Bacterial membranes: the effects of chill storage and food processing. An overview.

The shelf life of food is extended by refrigeration because the metabolic processes of food-associated microorganisms are slowed by the lowered temperature. Nonetheless, cold-adapted psychrotrophic food-poisoning and food-spoilage bacteria remain a concern because they possess cold-adapted proteins and membrane lipids that facilitate growth at low temperatures. The use of membrane-disrupting novel preservation techniques, such as ultrasound, high hydrostatic pressure or pulsed electric field, offer the potential for an extension of shelf life. This review considers the interacting and potentially synergistic effects of chill storage or mild heat treatment on membrane properties, with the disruptive effects of membrane-targeted physical treatments.

Bacteria↗

The Antarctic Psychrobacter sp. TAD1 has two cold-active glutamate dehydrogenases with different cofactor specificities. Characterisation of the NAD+-dependent enzyme.

Psychrobacter sp. TAD1 is a psychrotolerant bacterium from Antarctic frozen continental water that grows from 2 to 25 degrees C with optimal growth rate at 20 degrees C. The new isolate contains two glutamate dehydrogenases (GDH), differing in their cofactor specificities, subunit sizes and arrangements, and thermal properties. NADP+-dependent GDH is a hexamer of 47 kDa subunits and it is comparable to other hexameric GDHs of family-I from bacteria and lower eukaria. The NAD+-dependent enzyme, described in this communication, has a subunit weight of 160 kDa and belongs to the novel class of GDHs with large size subunits. The enzyme is a dimer; this oligomeric arrangement has not been reported previously for GDH. Both enzymes have an apparent optimum temperature for activity of approximately 20 degrees C, but their cold activities and thermal labilities are different. The NAD+-dependent enzyme is more cold active: at 10 C it retains 50% of its maximal activity, compared with 10% for the NADP+-dependent enzyme. The NADP+-dependent enzyme is more heat stable, losing only 10% activity after heating for 30 min, compared with 95% for the NAD+-dependent enzyme. It is concluded that in Psychrobacter sp. TAD1 not only does NAD+-dependent GDH have a novel subunit molecular weight and arrangement, but that its polypeptide chains are folded differently from those of NADP+-dependent GDH, providing different cold-active properties to the two enzymes.

Adaptation, Physiological↗

Citrate synthase and 2-methylcitrate synthase: structural, functional and evolutionary relationships.

Following the complete sequencing of the Escherichia coli genome, it has been shown that the proposed second citrate synthase of this organism, recently described by the authors, is in fact a 2-methylcitrate synthase that possesses citrate synthase activity as a minor component. Whereas the hexameric citrate synthase is constitutively produced, the 2-methylcitrate synthase is induced during growth on propionate, and the catabolism of propionate to succinate and pyruvate via 2-methylcitrate is proposed. The citrate synthases of the psychrotolerant eubacterium DS2-3R, and of the thermophilic archaea Thermoplasma acidophilum and Pyrococcus furiosus, are approximately 40% identical in sequence to the Escherichia coli 2-methylcitrate synthase and also possess 2-methylcitrate synthase activity. The data are discussed with respect to the structure, function and evolution of citrate synthase and 2-methylcitrate synthase.

Bacterial Proteins↗

Re-evaluation of the hypothesis that biodegradable surfactants stimulate surface attachment of competent bacteria.

The hypothesis that biodegradable surfactants stimulate the attachment of biodegradation-competent bacteria to surfaces has been re-evaluated using a variant of the surfactant-degrading bacterium Pseudomonas sp. DES1 designated Pseudomonas sp. DES2. This variant was identical to the parental strain in terms of its carbon-utilization patterns and alcohol dehydrogenase and alkylsulfatase complements (enzymes involved in surfactant biodegradation), but differed markedly in its growth characteristics when using sodium dodecyl triethoxysulfate or triethylene glycol dodecyl ether as secondary carbon sources. Pseudomonas sp. DES1 exhibited diauxie in these surfactant-based culture media in contrast to Pseudomonas sp. DES2, which exhibited single-phase growth. Pseudomonas sp. DES2 did not attach to river sediment in a microcosm system when challenged with a dose of either surfactant, although it did biodegrade the substrate. In contrast, Pseudomonas sp. DES1 attached to the river sediment whilst biodegrading the test substrate. It is concluded that the ether-scission system, which is responsible for primary biodegradation of both substrates, is deregulated in Pseudomonas sp. DES2 in contrast to that in Pseudomonas sp. DES1, and that, contrary, to a previous hypothesis, biodegradable surfactants do not necessarily stimulate the attachment of biodegradation-competent bacteria during their biodegradation.

Bacterial Adhesion↗

Ether-bond scission in the biodegradation of alcohol ethoxylate nonionic surfactants by Pseudomonas sp. strain SC25A.

Pseudomonas sp. strain SC25A, previously isolated for its ability to grow on alcohol ethoxylates (PEG dodecyl ethers) as sole source of carbon and energy, was shown to be capable of growth on the dodecyl ethers of mono-, di, tri- and octaethylene glycols. Comparative growth yields for this series of alcohol ethoxylate nonionic surfactants indicated that, whereas all of the carbon of monoethylene glycol dodecyl ether (MEGDE) was assimilable, only the alkyl chains were assimilated from the higher ethoxamers. These results are interpreted in terms of a primary biodegradation mechanism in which the scission of the dodecyl-ether bond is the first step. In the case of MEGDE this step separates the dodecyl chain from a C2 fragment, both of which are readily assimilable; for the higher ethoxamers, the assimilable dodecyl chain is accompanied by an ether-containing PEG derivative which would require further rounds of either scission before assimilation. Whole cells and cell extracts converted [1-14C]MEGDE initially and very rapidly to radiolabelled dodecanol. Disappearance of [14C]dodecaol was accompanied by production of [14C]dodecanal. [14C]Dodecanoic acid was present at relatively low concentrations throughout the incubation periods. [14C]Dodecan-1, 12-dioic acid was produced in significant quantities (up to 25% radiolabel), and the onset of its production coincided with the peak concentration of dodecanal, the disappearance of which mirrored the appearance of the dioic acid. Under anaerobic conditions in the presence of cell extracts, dodecanol (55% of radiolabel) and dodecanal (22%) accumulated rapidly from MEGDE, but there was little subsequent conversion to mono- or dicarboxylic acids. These results are interpreted in terms of a pathway initiated by dodecyl-ether cleavage to produce dodecanol, which is subsequently oxidized to dodecanal and dodecanoic acid. The formation of dodecan-1, 12-dioic acid, probably from dodecanal, may represent a means of harbouring carbon under non-growing conditions.

Biodegradation, Environmental↗