Growth, respiration and survival of Legionella pneumophila at high temperatures

The optimum temperature for multiplication of legionella strains in culture media is around 37°C. The effect of high temperatures on the growth of strains isolated from various environments is poorly known. We studied the growth (cell multiplication, respiration) of clinical and environmental Legionella pneumophila strains in liquid media at intervals of 0.5°C in the temperature range from 41.6 to 51.6°C using a temperature gradient incubator. Cell multiplication and CO₂ production decreased markedly with all the strains at temperatures above 44–45°C. CO₂ continued to be produced up to 51.6C even if cell multiplication generally stopped at around 48.4–50.0C. Thus, legionella retained its metabolic activity beyond the maximum temperature for cell multiplication. The CO₂ production per bacterial cell (metabolic quotient, qCO₂) increased with increasing temperature up to 45°C, whereafter it decreased, the turning point being almost at the same at which the rate of cell multiplication decreased. The difference in qCO₂ between the strains may reflect their different physiological capacities for tolerating high temperatures.     

Ultrastructure of Nitrosospira sp. X101 with crystalline outer membrane protein (COMP)

Abstract The outer membrane (OM) structure of Nitrosospira sp. X101 was studied by different electron microscopic techniques and SDS-PAGE. A crystalline outer membrane protein was visible in freeze-etched cells, occasionally seen also in the thin sectioned cells, but was difficult to see in a negatively-stained preparation. The lattice probably consists of large globular protein subunits with a hexagonal arrangement. The molecular weights of the major proteins in the cell envelope are 35 kDa, 40 kDa and 42 kDa.     

Mineralization of carbon and nitrogen in soil samples taken from three fertilized pine stands: Long-term effects

Seven years after fertilization the rate of CO₂ production in the soil samples taken from the organic horizons of a poor pine forest site (Calluna vulgaris site type), treated with urea or ammonium nitrate with lime, was lower than that in the unfertilized soil. The same trend was also observed in samples of theEmpetrum-Calluna site type 14 years after fertilization. In the more fertileVaccinium myrtillus site type these rapidly-soluble N fertilizers had a long-term enhancing effect on the production of CO₂. Apatite and biotite eliminated the decreasing effect of urea on the production of CO₂. One reason for this might be the long-term increase in soil pH caused by apatite and biotite, or their constituents (Ca, Mg, K, P). Nitroform (a slow-releasing N fertilizer) had no statistically significant effect on the production of CO₂ in soil samples from any of the forest types. Despite the high N mineralization in the samples from nitroform fertilized soils there was no nitrification, and the high content of total N indicated that after nitroform fertilization the losses of N were low.The correlation between the net mineralization values for C (CO₂ production) and N was poor. However, multiple linear regression analysis, which also took into account the effect of nutrients and pH, indicated that there was a link between the mineralization of C and N.     

Growth of legionella and other heterotrophic bacteria in a circulating cooling water system exposed to ultraviolet irradiation

The effect of ultraviolet irradiation on the growth and occurrence of legionella and other heterotrophic bacteria in a circulating cooling water system was studied. Water of the reservoir was circulated once in 28 h through a side-stream open channel u.v. radiator consisting of two lamps. Viable counts of legionellas and heterotrophic bacteria in water immediately after the u.v. treatment were 0—12 and 0·7—1·2% of those in the reservoir, respectively. U.v. irradiation increased the concentration of easily assimilable organic carbon. In the u.v. irradiated water samples incubated in the laboratory the viable counts of heterotrophic bacteria reached the counts in reservoir water within 5 d. The increase in viable counts was mainly due to reactivation of bacterialcells damaged by u.v. light, not because of bacterial multiplication. Despite u.v. irradiation the bacterial numbers in the reservoir water, including legionellas, did not decrease during the experimental period of 33 d. The main growth of bacteria in the reservoir occurred in biofilm and sediment, which were never exposed to u.v. irradiation.     

Availability and mobility of nutrients in acid forest soil treated with fast and slow-release nutrients

The effects of slow and fast-release fertilizers (P, K, Mg) on the movement and availability of nutrients in acid forest soil were studied. Fast-release superphosphate, potassium chloride and magnesium sulphate and slow-release apatite (P) and biotite (K, Mg) were applied alone or together with urea or urea+limestone. The nutrient content in the organic horizon was determined one growing season and three growing seasons after the application, and in the mineral layer after one growing season. The movement of nutrient ions in the organic horizon was studied by an ion exchange resin bag method during a 5-month period following application. The fast-release salts immediately increased the soluble P and exchangeable K and Mg contents in the organic and mineral soils and in the resin bags. After three growing seasons the effect of K application in the organic layer was non-detectable and that of P had clearly diminished. Apatite gradually increased soluble P content in the organic layer, but biotite had only a minor effect on the K and Mg contents. The nutrients from the fast-release fertilizers had clearly become available and mobile in the year of application and were thus susceptible to leaching. The rate of nutrient release from apatite and biotite is slower and the added nutrients are retained in the organic horizon. Slow-release compounds, like apatite and biotite, might be potential fertilizers for counteracting acidic deposition and subsequent nutrient losses.     

Effect of freezing of water samples on viable counts of environmental mycobacteria

The effect of prolonged storage on mycobacteria and other heterotrophic bacteria in brook water samples was examined by determination of viable counts from fresh samples and again after water concentrates had been stored in nutrient broth at —75°C for 15 months. The counts of mycobacteria were on average three times higher after storage (range of ratio 0·9–10·4). In contrast, the viable counts of other heterotrophic bacteria were reduced by 69%. The increase in mycobacterial counts was probably due to break-up of microcolonies or release of attached bacteria from particles. The possibility of cultivating mycobacteria from frozen samples is of practical help in large-scale field surveys.     

Comparison of some enrichment broths and growth media for the isolation of thermophilic campylobacters from surface water samples

Three different enrichment broths and two selective growth media were compared for isolating thermophilic campylobacters by combined membrane filtration and enrichment techniques from surface waters of different physical, chemical and bacteriological characteristics. Fifty-two strains of campylobacters were isolated from total of 1668 cultures. The various broth/medium combinations did not affect the dominance of C. jejuni over C. coli (total 49 C. jejuni and three C. coli). The most efficient combinations of enrichment broth and growth media were either Oosterom broth/blood-free charcoal-cefoperazone-deoxycholate agar (CCDA) medium or blood-free charcoal-cefoperazone-deoxycholate (CCD) broth/CCDA medium. Modified Preston broth (sheep blood instead of horse blood) with either of the growth media gave significantly lower yields although it suppressed efficiently the growth of contaminants. Skirrow medium had lower selectivity than CCDA medium and gave slightly lower isolation rate. Enrichment time (24 or 48 h) did not affect the isolation frequency of campylobacters but longer enrichment time increased the growth of contaminants. Prefiltration through membranes of pore sizes 5.0 and 1.2 microns decreased the growth of contaminants. However, these membranes retain campylobacters and must be cultured to avoid underestimation. From more polluted waters campylobacters were isolated most frequently with CCD broth and CCDA medium.     

Lakes as nitrous oxide sources in the boreal landscape

Estimates of regional and global freshwater N₂O emissions have remained inaccurate due to scarce data and complexity of the multiple processes driving N₂O fluxes the focus predominantly being on summer time measurements from emission hot spots, agricultural streams. Here, we present four‐season data of N₂O concentrations in the water columns of randomly selected boreal lakes covering a large variation in latitude, lake type, area, depth, water chemistry, and land use cover. Nitrate was the key driver for N₂O dynamics, explaining as much as 78% of the variation of the seasonal mean N₂O concentrations across all lakes. Nitrate concentrations varied among seasons being highest in winter and lowest in summer. Of the surface water samples, 71% were oversaturated with N₂O relative to the atmosphere. Largest oversaturation was measured in winter and lowest in summer stressing the importance to include full year N₂O measurements in annual emission estimates. Including winter data resulted in fourfold annual N₂O emission estimates compared to summer only measurements. Nutrient‐rich calcareous and large humic lakes had the highest annual N₂O emissions. Our emission estimates for Finnish and boreal lakes are 0.6 and 29 Gg N₂O‐N/year, respectively. The global warming potential of N₂O from lakes cannot be neglected in the boreal landscape, being 35% of that of diffusive CH₄ emission in Finnish lakes.     

Insect oviposition preference between Epichloë‐symbiotic and Epichloë‐free grasses does not necessarily reflect larval performance

Variation in plant communities is likely to modulate the feeding and oviposition behavior of herbivorous insects, and plant‐associated microbes are largely ignored in this context. Here, we take into account that insects feeding on grasses commonly encounter systemic and vertically transmitted (via seeds) fungal Epichloë endophytes, which are regarded as defensive grass mutualists. Defensive mutualism is primarily attributable to alkaloids of fungal origin. To study the effects of Epichloë on insect behavior and performance, we selected wild tall fescue (Festuca arundinacea) and red fescue (Festuca rubra) as grass–endophyte models. The plants used either harbored the systemic endophyte (E+) or were endophyte‐free (E?). As a model herbivore, we selected the Coenonympha hero butterfly feeding on grasses as larvae. We examined both oviposition and feeding preferences of the herbivore as well as larval performance in relation to the presence of Epichloë endophytes in the plants. Our findings did not clearly support the female's oviposition preference to reflect the performance of her offspring. First, the preference responses depended greatly on the grass–endophyte symbiotum. In F. arundinacea, C. hero females preferred E+ individuals in oviposition‐choice tests, whereas in F. rubra, the endophytes may decrease exploitation, as both C. hero adults and larvae preferred E? grasses. Second, the endophytes had no effect on larval performance. Overall, F. arundinacea was an inferior host for C. hero larvae. However, the attraction of C. hero females to E+ may not be maladaptive if these plants constitute a favorable oviposition substrate for reasons other than the plants' nutritional quality. For example, rougher surface of E+ plant may physically facilitate the attachment of eggs, or the plants offer greater protection from natural enemies. Our results highlight the importance of considering the preference of herbivorous insects in studies involving the endophyte‐symbiotic grasses as host plants.     

Juvenile parkinsonism,hypogonadism and Leigh-like MRI changes in a patient with m.4296G>A mutation in mitochondrial DNA

Leigh syndrome is a mitochondrial disease with considerable clinical and genetic variation. We present a 16-year-old boy with Leigh-like syndrome and broad developmental retardation, parkinsonism and hypogonadism. Sequencing of the entire mitochondrial DNA from blood revealed the m.4296G>A mutation in the MT-TI gene. The mutation was heteroplasmic with a 95% proportion of the mutant genome, while the proportion was 58% in the blood of the patient's clinically healthy mother. Our results suggest that m.4296G>A is pathogenic in humans, and that the phenotype related to this change includes Leigh-like syndrome in adolescence with parkinsonism and hypogonadism, in addition to the previously reported early infantile Leigh syndrome.