Dense Crithidia Growth and Heme Sparing: Relation to Fe,Cu, Mo Chelation*

SYNOPSIS. Heme, intrinsically required by Trypanosomatidae, is unstable, especially in conventional alkaline (pH 7.2–8.0) media. Low solubility of heme in a pH 6.5 basal medium (developed to assay biopterin with Crithidia fasciculata) posed a problem: in media acidified during growth because of glycolysis, heme precipitated, perhaps contributed to acid-limited growth and interfered with densitometric estimation of growth. The remedy was to: replace glucose with less rapidly metabolized mannitol; distribute media in thin layers to promote oxidation of acetate, fumarate, and malate (presumably leaving an alkaline residue); and buffer heavily with histidine + Good zwitterionic buffers, and superimpcse physiological buffering by arginine + asparagine whose catabolism appeared to yield an excess of NH⁺₄ over acid. Thereupon, Fe and Cu deficiencies sharply limited growth in the medium whose main chelators were: (a) 2,3–dihydroxybenzoic + 5-sulfosalicylic acids (which preferentially bind transitional elements at their higher valences; (b) malic and gluconic acids; and (c) histidine. With unconventionally heightened concentrations of Fe, Cu, and Mo (the latter serving as Cu buffer as well as nutrient per se), the hemin concentration could be lowered, widening the margin of safety for heme solubility. Growth then reached 1.4 × 10⁸ cell/ml. This medium may serve to screen for ligands promoting uptake or release of Fe and Cu. The increased growth is a step towards improving the assay medium for biopterin and practical use of Crithidia to assay several B vitamins and essential amino acids for metazoa.     

Isolation of polymorphic tetranucleotide microsatellite markers for the black‐bellied seedcracker (Pyrenestes ostrinus)

We have isolated 10 polymorphic microsatellite markers for the black‐bellied seedcracker (Pyrenestes ostrinus) from genomic libraries enriched either for (AAGG)_n or (ATCT)_n repetitive elements and characterized them in 39 individuals. The number of alleles ranged from two to 27 per locus with the observed heterozygosity ranging from 0.38 to 0.94. These markers will be useful for analysis of questions concerning parentage and population genetic structure.     

Biochemical and Cytological Evidence for an Overabundance of Mucocysts in the bcd Pattern Mutant of Tetrahymena thermophila

ABSTRACT. Three acidic proteins (42 kD, 43 kD and 50 kD) were present in unusually high concentrations in cortical preparations of the Tetrahymena pattern mutant broadened cortical domains (bcd). Antisera to the 42-kD and 50-kD proteins bound to discharging mucocysts and food vacuole contents in both wild-type and mutant cells. Subsequent analysis revealed that bcd mutant cell pellicles possess five times more "docked" mucocysts than their wild-type counterparts.     

A revision of the genus Naroma (Lepidoptera: Lymantriidae)

A key is given to the four species of Naroma now recognised. The species are redescribed and their distribution is listed. The early stages of N.varipes and N.signifera are described by D.G.Sevastopulo.     

Analysis of Genomic G + C Content, Codon Usage, Initiator Codon Context and Translation Termination Sites In Tetrahymena Thermophila

In recent years, the amount of molecular sequencing data from Tetrahymena thermophila has dramatically increased. We analyzed G + C content, codon usage, initiator codon context and stop codon sites in the extremely A + T rich genome of this ciliate. Average G + C content was 38% for protein coding regions, 21% for 5' non-coding sequences, 19% for 3' non-coding sequences, 15% for introns, 19% for micronuclear limited sequences and 17% for macronuclear retained sequences flanking micronuclear specific regions. The 75 available T. thermophila protein coding sequences favored codons ending in T and, where possible, avoided those with G in the third position. Highly expressed genes were relatively G + C-rich and exhibited an extremely biased pattern of codon usage while developmentally regulated genes were more A + T-rich and showed less codon usage bias. Regions immediately preceding Tetrahymena translation initiator codons were generally A-rich. For the 60 stop codons examined, the frequency of G in the end + 1 site was much higher than expected whereas C never occupied this position.     

A POLARLY FLAGELLATED MEMBER OF THE ENTEROBACTERIACEAE

SUMMARY: An organism which has been frequently isolated from the rhizosphere of plants of Lolium perenne (Perennial Rye-grass) is described; it has the typical morphology and flagellation of Pseudomonas but the biochemical reactions of Klebsiella.     

Hemispheric‐scale patterns of climate‐related shifts in planktonic diatoms from North American and European lakes

A synthesis of over 200 diatom‐based paleolimnological records from nonacidified/nonenriched lakes reveals remarkably similar taxon‐specific shifts across the Northern Hemisphere since the 19th century. Our data indicate that these diatom shifts occurred in conjunction with changes in freshwater habitat structure and quality, which, in turn, we link to hemispheric warming trends. Significant increases in the relative abundances of planktonic Cyclotella taxa (P<0.01) were concurrent with sharp declines in both heavily silicified Aulacoseira taxa (P<0.01) and benthic Fragilaria taxa (P<0.01). We demonstrate that this trend is not limited to Arctic and alpine environments, but that lakes at temperate latitudes are now showing similar ecological changes. As expected, the onset of biological responses to warming occurred significantly earlier (P<0.05) in climatically sensitive Arctic regions (median age=ad 1870) compared with temperate regions (median age=ad 1970). In a detailed paleolimnological case study, we report strong relationships (P<0.005) between sedimentary diatom data from Whitefish Bay, Lake of the Woods (Ontario, Canada), and long‐term changes in air temperature and ice‐out records. Other potential environmental factors, such as atmospheric nitrogen deposition, could not explain our observations. These data provide clear evidence that unparalleled warming over the last few decades resulted in substantial increases in the length of the ice‐free period that, similar to 19th century changes in high‐latitude lakes, likely triggered a reorganization of diatom community composition. We show that many nonacidified, nutrient‐poor, freshwater ecosystems throughout the Northern Hemisphere have crossed important climatically induced ecological thresholds. These findings are worrisome, as the ecological changes that we report at both mid‐ and high‐latitude sites have occurred with increases in mean annual air temperature that are less than half of what is projected for these regions over the next half century.     

Effects of experimental drought on soil respiration and radiocarbon efflux from a temperate forest soil

Soil moisture affects microbial decay of SOM and rhizosphere respiration (RR) in temperate forest soils, but isolating the response of soil respiration (SR) to summer drought and subsequent wetting is difficult because moisture changes are often confounded with temperature variation. We distinguished between temperature and moisture effects by simulation of prolonged soil droughts in a mixed deciduous forest at the Harvard Forest, Massachusetts. Roofs constructed over triplicate 5 × 5 m² plots excluded throughfall water during the summers of 2001 (168 mm) and 2002 (344 mm), while adjacent control plots received ambient throughfall and the same natural temperature regime. In 2003, throughfall was not excluded to assess the response of SR under natural weather conditions after two prolonged summer droughts. Throughfall exclusion significantly decreased mean SR rate by 53 mg C m^?2 h^?1 over 84 days in 2001, and by 68 mg C m^?2 h^?1 over 126 days in 2002, representing 10–30% of annual SR in this forest and 35–75% of annual net ecosystem exchange (NEE) of C. The differences in SR were best explained by differences in gravimetric water content in the Oi horizon (r²=0.69) and the Oe/Oa horizon (r²=0.60). Volumetric water content of the A horizon was not significantly affected by throughfall exclusion. The radiocarbon signature of soil CO₂ efflux and of CO₂ respired during incubations of O horizon, A horizon and living roots allowed partitioning of SR into contributions from young C substrate (including RR) and from decomposition of older SOM. RR (root respiration and microbial respiration of young substrates in the rhizosphere) made up 43–71% of the total C respired in the control plots and 41–80% in the exclusion plots, and tended to increase with drought. An exception to this trend was an interesting increase in CO₂ efflux of radiocarbon‐rich substrates during a period of abundant growth of mushrooms. Our results suggest that prolonged summer droughts decrease primarily heterotrophic respiration in the O horizon, which could cause increases in the storage of soil organic carbon in this forest. However, the C stored during two summers of simulated drought was only partly released as increased respiration during the following summer of natural throughfall. We do not know if this soil C sink during drought is transient or long lasting. In any case, differential decomposition of the O horizon caused by interannual variation of precipitation probably contributes significantly to observed interannual variation of NEE in temperate forests.