Phenanthrene degradation by estuarine surface microlayer and bulk water microbial populations

Paired surface microlayer and bulk water samples from five sites in the Great Bay Estuary, New Hampshire, were examined with regard to numbers of bacteria,¹⁴C-phenanthrene biodegradation potentials, and organic and inorganic chemical characteristics. Microlayer samples were generally enriched in nutrients (N and P), dissolved organic matter, and culturable heterotrophic bacteria compared with their corresponding bulk waters. Microlayer samples from marina environments were also enriched in aromatic hydrocarbons, as determined by UV spectrophotometric and fluorometric analyses, and demonstrated substantial phenanthrene biodegradation activity in the assay employed. Biodegradation activity of marina bulk water samples ranged from nil to levels exceeding those exhibited by microlayer samples. No diminution of biodegradation activity was observed after filtration (1.2 m effective retention) of microlayer water, indicating that the responsible organisms were not particle-associated. Phenanthrene-degrading bacteria, enumerated by counting clearing zones in a crystalline phenanthrene overlay after colony development on a phenanthrene/toluene agar (PTA) medium, were superior to epifluorescence direct counts or standard plate counts on PTA or estuarine nutrient agar in predicting¹⁴C-phenanthrene biodegradative activity.     

Factors regulating cladoceran dynamics in a Venezuelan floodplain lake

Five cladoceran zooplankton species in a Venezuelan floodplainlake, Laguna la Orsinera, were analyzed to identify factorscontrolling their dynamics over a period encompassing isolation,inundation by the Orinoco River and drainage. The abundanceof each species increased rapidly to a maximum associated withflooding of the lake by the Orinoco River. We calculated birth,death and population growth rates, and developed a method forestimating uncertainty in these parameters, in order to interpretobserved cladoceran abundance patterns. Rapid increases in populationsizes resulted both from high birth rates associated with decreasingturbidity following inundation and from the hatching of restingeggs. Birth rates remained high throughout the study, but highdeath rates restricted each population to a brief maximum andthen maintained small population sizes throughout the remainderof the study. Our demographic analyses suggest that high mortalityresulted from intense predation by fish and by Chaoborus larvae,rather than from resource limitation, and that this mortalitywas the major factor regulating cladoceran abundance duringinundation and drainage in this tropical floodplain lake.     

Production of poly-β-hydroxybutyrate by Azotobacter vinelandii strain UWD during growth on molasses and other complex carbon sources

Summary Azotobacter vinelandii strain UWD formed >2 mg/ml poly--hydroxybutyrate (pHB) during exponential growth in media containing ammonium acetate and 1% w/v glucose, fructose, sucrose, or maltose, and >1.5 mg/ml with 1% w/v sodium gluconate or glycerol. After acetate exhaustion, pHB formation accompanied carbohydrate utilization and pHB rapidly accounted for 53%–70% of the cell mass. Strain UWD also formed >2 mg/ml pHB when it was grown with 2% w/v corn syrup, cane molasses, beet molasses, or malt extract. Beet molasses had a growth stimulatory effect which promoted higher yields of pHB/ml and a high ratio of pHB/protein. Malt extract also promoted higher yields of pHB/ml. In this case, pHB formation was no longer subject to acetate repression and the cells contained a higher ratio of pHB/protein. This study shows that unrefined carbon sources support pHB formation in strain UWD and that the yields of pHB were comparable to or better than those obtained with refined carbon sources.     

Formation of Metallogenium-like structures by a manganese-oxiding fungus

Structures resembling Metallogenium spp. were observed in agar and in liquid cultures of a Mn-oxidizing basidiomycetous fungus only when Mn²⁺ was oxidized. Fungal viability was necessary for formation of the structures; Mn²⁺ concentration and the presence or absence of agar in the medium were important factors determining their morphology. Slide cultures revealed no identifiable cells in any stage of development. Fluorescent dyes that stained nucleic acids and polysaccharides in the fungal hyphae did not stain the Metallogenium-like structures. Likewise, Rhodamine 123, a fluorescent probe for membrane potential, stained fungal mitochondria, but did not stain the structures. Thin sections through the structures showed no biological membranes or other cellular features. Only the characteristic ultrastructure of biological Mn oxides were observed in serial thin sections. In agar, unfixed structures disappeared permanently during reduction of Mn oxides with hydroxylamine. Glutaraldehyde fixation stabilized these structures. Fixed structures lost most of their original phase density during reduction with hydroxylamine, but continuous microscopic observations showed that their phase density could be restored by staining with Coomassie blue. Structures that formed in liquid medium did not require stabilization with glutaraldehyde during reduction of Mn oxides. They, too, lost their original phase density during reduction with hydroxylamine; phase density could be restored by staining with cationic colloidal iron or Coomassie blue. The results suggest that the Metallogenium-like structures were formed as a result of Mn oxidation associated with exopolymers produced by the fungus.Non-standard abbreviations HEPES (N-hydroxyethylpiperazine-N-2-ethane sulfonic acid) - DAPI (4,6-diamidino-2-phenylindole) - PIPES (piperazine-N,N-bis[2-ethane sulfonic acid])     

Quararibea (Bombacaceae): Five new species from moist and wet forests of Costa Rica and Panama

Five new species ofQuararibea from costa Rica and Panama are described and illustrated, with notes on their ecology and relationships.Quararibea gomeziana, Q. pendula, andQ. santaritensis are from the Caribbean lowlands of the Provinces of Limón, Costa Rica, and of Bocas del Toro and Colón, Panama.Quararibea aurantiocalyx andQ. costaricensis are from montane habitats of Panama and Costa Rica. The exceptionally long pedicels ofQ. pendula far exceed those of other known members ofQuararibea and may prove to be the most striking example of adaptation to bat pollination and fruit dispersal in the genus.Quararibea costaricensis, a relatively common species, has long been erroneously identified asQ. platyphylla, a much rarer inhabitant of the same region.     

Catechol Formation and Melanization by Na+ -Dependent Azotobacter chroococcum: a Protective Mechanism for Aeroadaptation?

Aeroadaptive microaerophilic Azotobacter chroococcum 184 produced a cell-associated black pigment when grown at high aeration rates under nitrogen-fixing conditions. This pigment was shown to be a catechol melanin. Polyphenol oxidase activity was detected in cell extracts of cells grown for 72 h. Melanin formation was optimal in the later stages of growth, and there was no correlation between nitrogenase activity and melanization. Nitrogenase activity in strain 184 was optimal at 10% O₂, and melanin formation was suppressed by O₂ limitation. In the presence of charcoal, an adsorbent of toxic oxygen intermediates, and benzoic acid, a scavenger of hydroxyl radicals, melanization was inhibited. However, in the presence of copper, the intensity of pigment color increased and melanization was accelerated. Copper also eliminated catalase and peroxidase activities of the organism but still permitted aerobic growth. In the presence of low levels of iron, melanization was accelerated under high aeration rates, and under low rates of aeration, melanization was observed only at higher levels of iron. Hydroxamate-siderophore production was detectable in the presence of soluble iron under high rates of aeration but was repressed by the same levels of iron under low aeration rates. Unlike melanization and hydroxamate formation, catechol formation was observed under both low and high rates of aeration under nitrogen-fixing conditions. Catechol formation and melanization were repressed by 14 mM NH₄⁺, at which level nitrogenase activity was also repressed. Copper reversed the repressive effect of NH₄⁺. A role for catechol formation and melanization in aeroadaptation is proposed.     

Nutrient release rates from the sediments of Saginaw Bay,Lake Huron

Direct measurements of net production rates and pore water profiles of solutes in the fine-grained sediments of Saginaw Bay, imply corresponding steady-state fluxes to the overlying water of 1.1–1.3 (I), 450–1010 (NH₄ ⁺), 1250–2650 (Si(OH)₄), 3000–3400 (Ca²⁺), 440–1330 (Mg²⁺), 1.5–728 (Fe²⁺), and 179–281 (Mn²⁺) moles/m²/day and 11.0–11.8 (alkalinity) meq/m²/day at 17.5 °C. Silica production rates in sediments apparently follow first order kinetics with a rate coefficient of 0.09/day and a steady-state silica concentration of 1.2 mM at 23.5°C. The remaining solutes follow kinetics approximately independent of solute concentration over the range of concentrations observed. Measured solute production rates are consistent with observed solute profiles only if lateral diffusion gradients are maintained in the sediments by the burrowing and irrigation activity of benthic organisms such asChironomous, the dominant burrower in Saginaw Bay. Assuming that solute fluxes from Saginaw Bay are representative of all of the post-glacial sediments of Lake Huron, the iodine flux from sediments is comparable to the total fluvial input of iodine. The extrapolated silica fluxes from Lake Huron sediments balance the estimated biogenic silica flux to the sediments.     

Hepatic glycogen metabolism in the db/db mouse

Summary Knowledge of the metabolic changes that occur in insulin-resistant type 2 diabetes is relatively lacking compared to insulin-deficient type 1 diabetes. This paper summarizes the importance of the C57BL/KsJ-db/db mouse as a model of type 2 diabetes, and illustrates the effects that insulin-deficient and insulin-resistant states have on hepatic glycogen metabolism. A longitudinal study of db/db mice of ages 2–15 weeks revealed that significant changes in certain parameters of hepatic glycogen metabolism occur during this period. The liver glycogen levels were similar between diabetic and control mice. However, glycogen particles from db/db mice were on average smaller in mass and had shorter exterior and interior chain lengths. Total phosphorylase and phosphorylase a activities were elevated in the genetically diabetic mice. This was primarily due to an increase in the amount of enzymic protein apparently the result of a decreased rate of degradation. It was not possible to find a consistent alteration in glycogen synthase activity in the db/db mice. Glycogen synthase and phosphorylase from diabetic liver revealed some changes in kinetic properties in the form of a decrease in V_max, and altered sensitivity to inhibitors like ATP. The altered glycogen structure in db/db mice may have contributed to changes in the activities and properties of glycogen synthase and phosphorylase. The exact role played by hormones (insulin and glucagon) in these changes is not clear but further studies should reveal their contributions. The db/db mouse provides a good model for type 2 diabetes and for fluctuating insulin and glucagon ratios. Its use should clarify the regulation of hepatic glycogen metabolism and other metabolic processes known to be controlled by these hormones. The other animal models of type 2 diabetes, ob/ob mouse and fatty Zucker (fa/fa) rat, show similar impairment of hepatic glycogen metabolism. The concentrations of glycogen metabolizing enzymes are high and in vitro studies indicate enhanced rate of glycogen synthesis and breakdown. However, streptozotocin-induced diabetic animals and BB rats which resemble insulin-deficient type 1 diabetes are characterized by decreased glycogen turnover as a result of reduction in the levels of glycogen metabolizing enzymes.     

Comparative studies of hepatotoxicity and fumonisin B1 and B2 content of water and chloroform/methanol extracts of Fusarium moniliforme strain MRC 826 culture material

Fusarium moniliforme has been associated with several diseases including equine leukoencephalomalacia, human esophageal cancer and hepatotoxicity/hepatocarcinogenicity in laboratory animals. The potential health risks to animals and humans posed by F. moniliforme contaminated grains cannot be assessed until the toxins are identified and toxicologically evaluated. As part of a systematic approach to identifying the hepatotoxins produced by F. moniliforme, diets containing aqueous and chloroform/methanol (11) extracts of F. moniliforme strain MRC 826 culture material (CM) and/or the extracted CM residues were fed to male Sprague-Dawley rats for four weeks. Serum alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase activities were increased after two and four weeks and microscopic liver lesions were found in those animals fed aqueous CM extract and the CM residue after chloroform/ methanol extraction. Fumonisins B₁ and B₂ were extracted from the CM by water, but not chloroform/ methanol, and were present in the toxic diets at concentrations of 93–139 and 82–147 ppm, respectively. Nontoxic diets contained 22 ppm fumonisin B₁ and 65 ppm fumonisin B₂.Abbreviations CM culture material - ELEM equine leukoencephalomalacia Mention of a trademark, proprietory name or vendor does not imply its approval by the US Department of Agriculture to the exclusion of others that may also be suitable.