Long-term changes of diatoms and chemistry in headwater streams polluted by atmospheric deposition of sulphur and nitrogen compounds

1. Samples for the analysis of attached diatoms and surface water chemistry were taken at thirty-five stations in sixteen soft-water streams in the Netherlands during autumn 1990. Most were located in pine forest and heathland catchments, fed by deep groundwater (median residence time 87 years), with no direct influence of agricultural or urban drainage water. 2. The chemical data from sixteen stations were compared with data collected in 1974 and 1981. There were no significant increases in pH (6.3 in 1974, 6–4 in 1990) or in sulphate (200 mmol m^?3 in 1974, 229 mmol m^?3 in 1990). Over the same period nitrate increased significantly from 17 to 158 mmol m^?3, while alkalinity decreased significantly from 355 to 251 meq m^?3. 3. The most important correlates with the distribution of diatoms, as revealed by canonical correspondence analysis, were pH, nitrate, cross-sectional area of the stream and the Ca/(Ca + Cl) ratio. 4. Diatom assemblages of upstream stations indicated more acid conditions than those of downstream stations. Also there was considerable variation in the diatom assemblages of different habitats, 5. A diatom-pH transfer function was developed using the weighted-averaging method. The function was applied to comparable pairs of samples which were collected in 1974 and 1990 at ten stations. There was a significant decrease in the median diatom-inferred pH from 6.78 in 1974 to 6.55 in 1990. 6. The diatom assemblages are diverse and contain a high proportion of filamentous diatoms, particularly Aulacoseira crenulata and A. alpigena, which are rare in Western Europe. This is a response to the low current velocity (median 5 cm^?1) and the relatively constant deep groundwater discharge.     

Phytosterols and Other Lipids as Survival Factors for Tetrahymena at 0–5 C

SYNOPSIS. Tetrahymena pyriformis syngen 1, mating type II, (optimal growth temperature ∼ 37 C) ordinarily dies out in 5-14 days at 0-5 C. Dying cells were lumpy, suggesting membrane damage. By supplying crude soy lecithin, survival at 0-5 C was prolonged (after growth in peptone-yeast-dextrin) to at least 22 weeks. Crude soy sterols or sitosterol or stigmasterol, and antioxidant, e.g., Ionox 330 or ascorbylpalmitate, permitted survival of cells in suspension or in growth media for at least 16-22 weeks. These sterols are known to protect against triparanol toxicity, which suggested that triparanol, which blocks cholesterol synthesis in higher animals, might enhance cold-induced injury. Triparanol was more toxic at 0–5 than at 28 C for cell suspensions and cells in growth medium; this toxicity was annulled by crude soy lecithin or β-sitosterol, the only phytosterol tested. The synthetic medium intended as a control on the crude media became toxic at 0–5 C. Protection against cold damage is discussed as a means of elucidating the role of sterols—especially phytosterols—and other lipids in maintaining the integrity of the ciliate cell membrane.     

Growth of Tetrahymena in Carbohydate-Free High-Glutamate Media

SYNOPSIS. In chemically defined media, pH 6.4–6.8, supplementation with glycerol permitted growth equal to that with a carbohydrate supplement. In a pH 5.4 medium described in detail, containing L-glutamic acid 0.25%, supplementation with L-serine (essential for this strain), especially when further supplemented with proline, likewise permitted growth nearly equal to that with carbohydrate. These results are discussed in relation to the question of which components of peptone act as substrates and the resemblances of the Tetrahymena pattern of substrate utilization to that of metazoa.     

A multiplex set of microsatellite markers for the scarlet rosefinch (Carpodacus erythrinus)

Cardueline finches have become important models in studies of sexual selection and evolution of carotenoid‐based ornamentation. Here, we describe eight new polymorphic microsatellites isolated from the Scarlet rosefinch (Carpodacus erythrinus) and four from the House finch (Carpodacus mexicanus). Together with the cross‐species amplification of additional loci, originally published for two species of songbirds, we optimized a multiplex panel for C. erythrinus allowing genotyping of 22 polymorphic loci. Number of alleles and heterozygosity per locus in a sample of 34 individuals ranged from three to 38 and from 0.27 to 0.94, respectively.     

Immunologic-mediated Protection of Trypanosoma congolense-Infected Mice by Polyribonucleotides

SYNOPSIS. When the synthetic polyribonucleotides polyinosinic acid-polycytidylic acid (poly I poly C) and polyadenylic acid-polyuridylic acid (poly A poly U) were tested against mice infected with varying numbers of Trypanosoma congolense the results varied with the method of passage of trypanosomes in mice. Thus, when 100 flagellates were passaged every 7th day and experiments were initiated with these trypanosomes from mice on the 7th day of their infection, the protective effects of poly I poly C and poly A poly U apparently varied independently of each other as assayed by the mean parasitemias and cumulative mortalities of infected mice. Poly I poly C-resistant and poly I poly C-susceptible variants (“R” and “S”, respectively) were isolated and maintained in mice by passage of 10⁶ trypanosomes every 4th day. Mice infected with these variants responded consistently to poly I poly C and poly A poly U injections in that mice infected with the “R” variant showed no response to either polyribonucleotide but those infected with the “S” variant consistently had a decrease in mean parasitemias and cumulative mortality when treated with poly I poly C, but not with poly A poly U. Using mice infected with the “S” variant, the protective effect of poly I poly C was dose-dependent and best protection was afforded when 1st injections of poly I poly C were given around the time of infection of the mice. The protective effects of the synthetic polyribonucleotides used in these experiments are probably due to their immunologic enhancing capacities and not to interferon. To support this, injections of Newcastle disease virus, a strong inducer of interferon in mice, did not protect against T. congolense in mice. It was not possible to determine whether serum from poly I poly C-treated mice had a greater neutralizing effect upon trypanosomes in vitro than serum from saline-treated mice since any effect of antibody was masked by a naturally occurring inhibitor in normal mouse serum which was reduced during infection. The protective effects of poly I poly C in T. congolense-infected mice were reversed by treatment with cyclophosphamide. This strong immunosuppressant, however, could not reverse the protective effects of poly I poly C against mice infected with Semliki Forest virus, strongly suggesting that the protective mechanisms stimulated by poly I poly C in these 2 infections were different. The response of mice infected with the “R” and “S” variants of T. congolense to synthetic polyribonucleotides is discussed in relation to antigenic variation of trypanosomes.     

Cryoprotectants for Crithidia fasciculata Stored at -20 C, with Notes on Trypanosoma gambiense and T. conorhini

SYNOPSIS. Cryoprotectants were tested in both complex and semidefined media for the trypanosomatid Crithidia fasciculata. Near log-phase or end-of-log-phase cultures were frozen for 24–48 hr at ∼ -20 C, then warmed in air to room temperature. Immediate motility was correlated with viability. The best protectant of the 83 tested was glycerol at ∼ 10% (w/v). Survival without cryoprotectant was rare. Outstanding cryoprotectants (perhaps also useful solvents for drugs poorly soluble in water) were: ethylene glycol; 2,2'-dioxyethanol (diethylene glycol); 1,2,4-butanetriol; 1,4-cyclohexanediol; dimethylsulfoxide; propylene glycol; and N -acetylethanolamine. Several sugars were active, e.g., D-arabinose, sucrose, and sorbitol. Trypanosomes tolerated cryoprotectants much less; tolerance was better in growth media than in suspension media. Trypanosoma gambiense was grown in blood-enriched media + 2-2.5% glycerol, suspended in 20% (w/v) glycerol. then frozen; this permitted 3-week survival. T. conorhini survived 4 weeks after growth in media containing glycerol 2.5%+ ethylene glycol 4%+ rutin 1.0 mg per 100 ml.