Influence of environmental parameters on polyphosphate accumulation inAcinetobacter sp.

The regulation of and the optimum conditions for polyphosphate accumulation inAcinetobacter sp. were determined.Acinetobacter strain 210A accumulated polyphosphate in the presence of an intra- or extracellular energy source. The accumulation of polyphosphate during endogenous respiration was stimulated by streptomycin and inhibited by KCN. The highest amount of polyphosphate was found in cells in which energy supply was not limited, namely at low growth rates under sulphur limitation, and in the stationary phase of growth when either the nitrogen or the sulphur source was depleted. The phosphorus accumulation was not affected by the pH between 6.5 and 9. There was a pronounced effect of the temperature on phosphorus accumulation but is varied from strain to strain.Acinetobacter strain 210A accumulated more phosphate at low temperatures, strain B8 showed an optimum accumulation at 27.5° C, while strain P accumulated phosphorus independently of the temperature. The optimum temperature for growth ofAcinetobacter strains tested ranged from 25 to 33° C, and the optimum pH was between 6 and 9.     

Exposure of Fischerella [Mastigocladus] to high and low temperature extremes: strain evaluation for a thermal mitigation process

In conjunction with a proposed algal cultivation scheme utilizing thermal effluent, twelve Fischerella strains were tested for tolerance to temperatures above and below their growth range. Exposure to 65 °C or 70 °C for 30 min caused bleaching and death of most or all cells. Effects of 60 °C exposure for periods of up to 2 h ranged from undetectable to severe for the various strains. Chlorophyll a content typically decreased 21–22% immediately following 60 °C or 65 °C (1 h) exposure. However, the 60 °C-shocked cultures regained normal Chl a content after 24 h at 45 °C, whereas Chl a in 65 °C-shocked cultures immediately lost visible autofluorescence and was later degraded. Exposure to 15 °C virtually stopped growth of all strains during a 48 h exposure period. Most strains grew as rapidly as 45 °C controls when restored to 45 °C, while a few strains recovered more slowly. Comparison with dark-incubated controls indicated that photooxidative damage did not occur during cold shock. Certain strains exhibited relatively rapid recovery from both heat and cold exposure, thus meeting the temperature tolerance criteria for the proposed algal cultivation process.     

Effect of the composition of the growth medium on morphology and reproduction ofAzotobacter chroococcum

Summary The growth of threeAzotobacter chroococcum strains was investigated in a number of media. The cultures were incubated at 29°C. and studied by means of a Wild phase contrast microscope. The experiments have clearly demonstrated that the morphology and reproduction ofA. chroococcum are influenced greatly on the one hand by conditions of nutrition and on the other hand also by the properties of the strains. Although all strains used in this work were able to produce gonidia and regenerative rods, these were not found in all media. The formation of filamentous cells, and vegetative cells inside such cells, depended also on the medium and, in some cases, on the strain. Cysts could sometimes resist a temperature of 50°C. for 5.5 minutes, but not a single strain with cysts survived temperatures of 60°C. or higher. Spores could not be detected by the usual staining methods.     

Enrichment, isolation and characterization of dialkyl sulfosuccinate degrading bacteriaComamonas terrigena N3H andComamonas terrigena N1C

Microorganisms derived from soil, sludge, sewage or river water contaminated by detergents or mineral oils were enriched and tested for the capability of primary biodegradation of dialkyl sulfosuccinates (DASS). Adapted mixed cultures, containing predominantly Gram-negative bacteria, degraded di-n-hexyl sulfosuccinate (DHSS) more readily than di-n-octyl sulfosuccinate (DOSS). Bacterial isolates obtained from enriched mixed cultures that show the highest rates of primary biodegradation and assimilation of DASS belong to the genusComamonas. Under nongrowth conditions, the DHSS primary biodegradation by these strains follows the 1st order reaction kinetics. The rate constants of primary degradation of DHSS by resting cells ofC. terrigena N3H were found to be 30 times higher than when catalyzed byC. terrigena N1C. The capacity for accelerated removal of surface-active properties of DHSS byC. terrigena N3H was retained within the range of pH 4–10 and at temperatures up to 55°C, while DHSS degradation byC. terrigena N1C was completely inhibited above 48°C. Temperature and pH limits for DHSS primary degradation by both strains were to a great extent wider than those for the viability of the cultures. Biodegradation capacity and metabolic stability ofC. terrigena N3H make this strain interesting from both scientific and technological point of view.     

Effects of temperature and food level on growth and development of a planktonic water mite

We analysed the relative effects of food availability and temperature on rates of growth and development of a predatory planktonic water mite, Piona exigua. Growth in length of mites fed Daphnia, Ceriodaphnia and Chydorus was analysed by Gompertz or von Bertalanffy curves; these curves were compared by parallel curve analysis. Growth rates of nymphs and adult female mites increased with temperature; the duration of the imagochrysalis stage decreased. Females grown at 10 °C were smaller at final size than females grown at 15 °C, 18 °C or 22 °C. Females reared at food levels of 15 or 30 prey l⁻¹ grew more slowly and were smaller than those provided with 60 or 120 prey l⁻¹. Nymphs grew more slowly when Daphnia were the only prey, than when smaller prey were available. Food level did not affect nymph growth at 10 °C or 15 °C, but growth at 18 °C or 22 °C may have been slowed at the lowest food levels. Synergistic effects of temperature and food level on nymph growth were apparent only from analysis of growth curves and not from stage duration data.     

Fruit-body production of test cultures ofFlammulina velutipes preserved for seven years by freezing at three different temperatures

Two strains ofFlammulina velutipes were cultured on PDA plates, and mycelial disks punched out using a cork borer were used for preservation. Five disks of a strain were put into a vial containing one of three cryoprotectants, 10% glycerol, 5% DMSO or 10% polyethylene glycol. Vials were then stored for 7 yr at −20°C, −85°C or liquid nitrogen temperature. The mycelial growth on PDA plates of the cryopreserved mycelial disks, as well as the usual subcultures, were tested two times. After the second test, spawns were prepared for fruit-body production tests by bottle cultivation from selected plates of the second growth tests. The yields of fruit-bodies varied among the cultures derived from the mycelial disks of the same strain preserved under different conditions. Variation in yields was observed even among the mycelial disks preserved at liquid nitrogen temperature, although the range of yield variation was narrower. The yield variation was obvious for the cultures which showed large retardation in the growth test. Four mycelial disks out of the six preserved at −20°C showed higher yields than those preserved at other temperatures. Among the cultures derived from strain FMC224, the control cultures preserved by subculture showed the lowest yield.     

Plant regeneration from microspore-derived embryos ofBrassica Napus: Effect of embryo age,culture temperature,osmotic pressure,and abscisic acid

Summary Microscope cultures ofBrassica napus cv. Topas undergo high frequency embryogenesis in vitro; however, the majority of microspore-derived embryos do not develop directly into plants but usually undergo abnormal development including the formation of secondary embryos on the hypocotyls. The present studies show that older embryos or embryos cultured at higher temperature (25° C) were more likely to undergo secondary embryogenesis whereas embryos cultured at 20° C or pretreated at 5° to 10° C for 28 days developed more readily into normal plants. Compared with embryos cultured at 25° C, those cultured at 20° C gave a threefold increase in normal plant production. Pretreatments at cooler temperatures (5° to 10° C) resulted in an additional two-to threefold increase in the recovery of normal plants. Higher osmoticum during pretreatment improved embryo survival at low temperatures but generally inhibited normal plant development. Abscisic acid was ineffective or deleterious.     

Selection of a Photosynthetic Bacterium Suitable for Hydrogen Production in Outdoor Cultures among Strains Isolated in the Seoul,Taegu, Sendai and Bangkok Areas

Very efficient hydrogen producing photosynthetic bacteria, strains SL1, SL3, SL16 and TG28 newly isolated in Korea, and strain KM113 newly isolated in the Sendai area, were found to be Rhodopseudomonas spp. To examine the stability of cell suspensions of the cultures for hydrogen production, which is closely associated with light absorption, we conducted larger scale cultures under periodic illumination (12-hr intervals) without stirring at 30°C using strains SL1 and Rhodopseudomonas sphaeroides B5, the latter was isolated in the Bangkok area. Both strains gave homogeneous cell suspensions throughout the incubation period and larger amounts of hydrogen were produced in a shorter period of time by both cultures than obtained with Rhodopseudomonas sp. TN3, an isolate from the Sendai area which was reported previously. With the cells of the new isolates and strains TN3 and B5 grown on glutamate-malate medium at 30°C, we measured hydrogen production at 20, 30 and 40°C in the same medium. Among them, strains SL1, SL16 and KM113 showed the highest hydrogen production activity at 30°C. The maximum hydrogen production rates with these strains were over 130 µ1/hr/mg dry cells, but at 40°C, the highest activity (138 µl/hr/mg dry cells) was obtained with strain B5. Since strain B5 also showed good activities at 20 and 30°C, we suggest that this strain might be suitable for hydrogen production in outdoor cultures.     

Field evaluations of systemic fungicides and derivatives of dithiocarbamic acid for the control of clubroot

Strains of Botrytis cinerea and Mucor mucedo germinated and grew over the range 0.25^°C. There were differences in germination rates and growth rates between strains of B. cinerea at any given temperature. Five of the benomyl-resistant strains germinated and grew more slowly than any of the other benomyl-resistant or benomyl-sensitive strains of B. cinerea tested. Strains of Rhizopus stolonifer and R. sexualis germinated and grew between 5 and 25^°C, and although some spores germinated at 2^°C, subsequent growth of the germ tubes and growth from a mycelial inoculum did not occur. Neither species germinated or grew at o^°C. The effect of temperature on mycelial growth in vitro was consistent with the ability of the strains of the four species to infect strawberry fruits.     

General introduction to the lists of threatened biotopes,flora and fauna of the trilateral Wadden Sea Area (red data book)

The effect of the acclimation temperature on the temperature tolerance ofPorphyra leucosticta, and on the temperature requirements for growth and survival ofEnteromorpha linza was determined under laboratory conditions. Thalli ofP. leucosticta (blade or Conchocelis phases), acclimated to twenty-five degrees, survived up to 30°C, i.e. 2°C more than those acclimated to 15°C which survived up to 28°C. Lower temperature tolerance of bothPorphyra phases that were acclimated to 15°C was −1°C after an 8-week exposure time at the experimental temperatures. The upper temperature tolerance ofE. linza also increased by 2°C, i.e. from 31 to 33°C, when it was acclimated to 30°C instead of 15°C. The lower temperature tolerance increased from 1 to −1°C, when it was acclimated to 5°C instead of 15°C.E. linza thalli acclimated for 4 weeks to 5 or 10°C reached their maximum growth at 15°C, i.e. at a 5°C lower temperature than those acclimated to 15 or 30°C. These thalli achieved higher growth rates in percent of maximal growth at low temperatures than those acclimated to 15 or 30°C. Thalli acclimated for 1 week to 5°C reached their maximum growth rate at 20°C and achieved growth rates at low temperatures similar to those recorded for thalli acclimated to 15°C. Thalli ofE. linza acclimated for 4 weeks to 5°C lost this acclimation after being post-cultivated for the same period at 15°C. That was not the case with thalli acclimated for 8 weeks to 5°C and post-acclimated for 4 weeks to 15°C. These thalli displayed similar growth patterns at 10–25°C, while a decline of growth rate was observed at 5 or 30°C. The significance of the acclimation potential ofE. linza with regard to its seasonality in the Gulf of Thessaloniki, and its distribution in the N Atlantic, is also discussed.     

A rapid identification method for aflatoxin-producing strains ofAspergillus flavus andA. parasiticus by ammonia vapor

The colony reverse of aflatoxin (AF)-producing strains ofAspergillus flavus andA. parasiticus turned pink when their cultures were exposed to ammonia vapor. The color change was visible for colonies grown on media suitable for AF production such as potato dextrose, coconut, and yeast extract sucrose agars after 2 d incubation at 25°C. Of the 120 strains ofA. flavus, A. parasiticus, and two related species inA. flavus group:A. oryzae andA. sojae tested in this study, only the AF-producing strains ofA. flavus andA. parasiticus showed the pink pigmentation. The color change occurred immediately after the colony was contacted with ammonia vapor. This method was useful for rapid screening the AF-producing strains ofA. flavus andA. parasiticus.     

TEMPERATURE AND IRRADIANCE EFFECTS ON GROWTH OF PITHOPHORA OEDOGONIA (CHLOROPHYCEAE) AND SPIROGYRA SP. (CHAROPHYCEAE)1

Growth responses of Pithophora oedogonia (Mont.) Wittr. and Spirogyra sp. to nine combinations of temperature (15°, 25°, and 35°C) and photon flux rate (50, 100, and 500 μmol·m^?2·s^?1) were determined using a three-factorial design. Maximum growth rates were measured at 35°C and 500 pmol·m^?2·s^?1 for P. oedogonia (0.247 d^?1) and 25°C and 500 μmol·m^?2·s^?1 for Spirogyra sp. (0.224 d^?1). Growth rates of P. oedogonia were strongly inhibited at 15°C (average decrease= 89%of maximum rate), indicating that this species is warm stenothermal. Growth rates of Spirogyra sp. were only moderately inhibited at 15° and 35°C (average decrease = 36 and 30%, respectively), suggesting that this species is eurythermal over the temperature range employed. Photon flux rate had a greater influence on growth of Spirogyra sp. (31% reduction at 50 pmol·m^?2·s^?1 and 25°C) than it did on growth of P. oedogonia (16% reduction at 50 μmol·m^?2·s^?1 and 35°C). Spirogyra sp. also exhibited much greater adjustments to its content of chlorophyll a (0.22–3.34 μg·mg fwt^?1) than did P. oedogonia (1.35–3.08 μg·mg fwt^?1). The chlorophyll a content of Spirogyra sp. increased in response to both reductions in photon flux rate and high temperatures (35°C). Observed species differences are discussed with respect to in situ patterns of seasonal abundance in Surrey Lake, Indiana, the effect of algal mat anatomy on the internal light environment, and the process of acclimation to changes in temperature and irradiance conditions.     

Effect of salinity and pH on growth and agar yield of Gracilaria tenuistipitata var. liui in laboratory and outdoor cultivation

Acclimation responses of the red alga Gracilaria tenuistipitata var. liui collected on the northwest coast of Philippines were determined in laboratory setups and outdoor cultivation tanks in Haifa, Israel. Growth under laboratory conditions was influenced by all three variables studied, namely, temperature (20 or 30 ^°C), salinity (20, 30 or39‰) and seawater pH (6.5, 7.0, 8.0 or ≥ 9.0). In 250 mL flasks lacking pH control growth was influenced by temperature only at 20 ‰, whereas at 39 ‰, growth rates were similar at 20 or 30 ^°C. In 500 mL cylinders in which pH was controlled, growth rates were significantly different at a pH of 6.5 and 7.0 for all salinities, with maximal rates occurring in 39 ‰. At pH 8.0, and above, growth rates between salinities were similar and reduced to approximately 50% at a pH of 9.0 compared to rates at a pH of 6.5. Photosynthesis responses generally resembled growth responses both, in 250 mL and 500 mL cultures. In 40-L outdoor tanks, weekly growth and agar yields were apparently enhanced by increasing light intensities (up to full sunlight) and nutrient concentrations (up to 0.2 mM PO₃ ^2- and 2.0 mM NH₄ ⁺), and rates averaged four times higher than rates determined in the smaller flask cultures. This study shows broad salinity tolerance of G. tenuistipitata var. liui and its ability to sustain growth rates that are among the highest measured for Gracilaria spp. in outdoor cultures. This revised version was published online in August 2006 with corrections to the Cover Date.     

A vitamin-free minimal synthetic medium forCryptococcus neoformans

The use of a simple synthetic medium is essential for study on the growth and physiology ofCryptococcus neoformans. In the present study, a minimal synthetic liquid medium (MSM) was tested for the growth of 23C. neoformans strains. This medium contained a low concentration of glucose, ammonium sulphate and inorganic salts with a pH value of 4.5, but no amino acids or vitamins. The strains were starved for 4 days to eliminate nutrients which might have been carried over from their pre-culture medium. Then, they were inoculated in the MSM at an initial OD of 0.020 at 550 nm and incubated at 37 °C for 20 days. Cell growth was generally monitored daily by measuring the absorbance at 550 nm. The medium supported the growth of the strains tested and gave an average final OD of 0.500. The results obtained indicate thatC. neoformans may be autotrophic with respect to vitamins and in particular to thiamine. The MSM medium is easy to prepare and store. It is highly reproducible and useful for studies on the growth and physiology ofC. neoformans.     

Soil temperature effects on competitiveness and growth ofRhizobium japonicum and on Rhizobium-induced chlorosis of soybeans

Summary The effects of temperature on growth in broth and soil and on competition for nodule formation betweenRhizobium japonicum serotypes USDA 76 and 94 compared to 6 and 110 were studied. Increasing root temperatures of Lee soybean from 20 to 35°C increased the competitiveness of 76 and 94 relative to 6 and 110 for all inoculum ratios such that at 30 and 35°C symptoms ofRhizobium-induced chlorosis appeared. Tolerance to elevated temperatures was exhibited by 76 and 110, but not 94 and 6 in broth and soil which suggested that increased competitiveness of 76 and 94 at high soil temperatures was not dependent upon growth at elevated temperatures. Nodulation and vegetative growth of Lee soybeans were at a minimum at 20°C and optimum at 30°C. Differences in competitiveness of 6 to previous studies indicated the need to standardize temperatures of assays. Differences in growth responses of 76 and 94 to temperature from a previous study suggested a confounding effect on different carbon sources in growth media. Scientific Article No. A-3721 Contribution No. 6697 of the Maryland Agric Exp Sta, Dept of Agronomy, College Park, MD 20742 and the USDA, ARS, Beltsville, MD 20705. Part of a thesis submitted by the senior author in partial fulfillment of the requirements for the M.S. Degree.     

Metarhizium spp. isolates from madagascar: Morphology and effect of high temperature on growth and infectivity to the migratory locust,Locusta migratoria

Five strains ofMetarhizium anisopliae (Metsch.) Sorokin and one strain ofMetarhizium flavoviride Gams &; Rozsypal originally isolated in Madagascar were studied. Measurements of conidia and, for the first time, also of blastospores produced in a liquid medium were used for species and variety determination. Blastospores ofM. flavoviride were more homogenous in their size than those ofM. anisopliae. Growth at high temperatures between 25° and 40°C showed that 4 isolates ofM. anisopliae grew at 36°C andM. flavoviride grew at 38°C. Using alternating day/night temperatures (8/16 h) the three strains tested could also tolerate 40°/25°C. In bioassays, fiveMetarhizium spp. isolates were tested against third and fourth instar larvae ofLocusta migratoria (L.) at two alternating day/night temperatures of 30°/25°C and 36°/25°C. In the cooler regime, all strains caused a mortality of 50% within 5.9 to 8.5 days (median lethal time), while in the 36°/25°C treatment only the thermophilicM. flavoviride and oneM. anisopliae strain isolated from a soil sample gave comparable results with median lethal times of 6.8 and 7.3 days, respectively.     

Mycelium growth kinetics and optimal temperature conditions for the cultivation of edible mushroom species on lignocellulosic substrates

The influence of environmental parameters on mycelial linear growth ofPleurotus ostreatus, P. eryngii, P. pulmonarius, Agrocybe aegerita, Lentinula edodes, Volvariella volvacea andAuricularia auricula-judae was determined in two different nutrient media in a wide range of temperature, forming the basis for the assessment of their temperature optimaV. volvacea grew faster at 35°C,P. eryngii at 25°C,P. ostreatus andP. pulmonarius at 30°C,A. aegerita at 25 or 30°C andA. auricula-judae at 20 or 25°C depending on the nutrient medium used andL. edodes at 20 or 30°C depending on the strain examined. The mycelium extension rates were evaluated on seven mushroom cultivation substrates: wheat straw, cotton gin-trash, peanut shells, poplar sawdust, oak sawdust, corn cobs and olive press-cake. The mycelium extension rates (linear growth and colonization rates) were determined by the ‘race-tube’ technique, and were found to be the highest on cotton gin-trash, peanut shells and poplar sawdust forPleurotus spp. andA. aegerita. Wheat straw, peanut shells and particularly cotton gin-trash supported fast growth ofV. volvacea, whereas wheat straw was the most suitable substrate forL. edodes andA. auricula-judae. Supplemented oak sawdust and olive press-cake were poor substrates for most species examined, white almost all strains performed adequately on corn cobs.     

Inhibition of methanogenesis and its reversal during biogas formation from cattle manure

The composition of volatile fatty acids in the biogas digester based on cattle manure as substrate and stabilised at 25°C showed that it contained 87–88% branched chain fatty acids, comprising of isobutyric and isovaleric acids, in comparison to 38 % observed in the digester operating at 35°C. Mixed cellulolytic cultures equilibrated at 25°C (C-25) and 35‡C (C-35) showed similar properties, but rates of hydrolysis were three times higher than that observed in a standard biogas digester. The proportion of isobutyric and isovaleric were drastically reduced when C-25 was grown with glucose or filter paper as substrates. The volatile fatty acids recovered from C-25 (at 25°C) inhibited growth of methanogens on acetate, whereas that from C-35 was not inhibitory. The inhibitory effects were due to the branched chain fatty acids and were observed with isobutyric acid at concentrations as low as 50 ppm. Addition of another micro-organismRhodotorula selected for growth on isobutyric completely reversed this inhibition. Results indicate that the aceticlastic methanogens are very sensitive to inhibition by branched chain fatty acids and reduction in methane formation in biogas digester at lower temperature may be due to this effect.     

Anatoxin-a concentration inAnabaena andAphanizomenon under different environmental conditions and comparison of growth by toxic and non-toxicAnabaena-strains — a laboratory study

Anatoxin-a-concentration in cells ofAnabaena- andAphanizomenon-strains and in their growth media were studied in the laboratory in batch cultures at different temperatures, light fluxes, orthophosphate and nitrate concentrations and with different nitrogen sources for growth. Toxin concentrations were detected by HPLC. Also, the growth of the toxicAnabaena-strains was compared to that of a non-toxic one. The non-toxicAnabaena was never found to produce anatoxin-a. The amount of toxin in the cells of the toxic strains was high, often exceeding 1% of their dry weight. High temperature decreased the amount of the toxin regardless of growth. Growth limiting low and growth inhibiting high light decreased the amount of the toxin in the cells ofAnabaena-strains. The highest light flux studied did not limit the growth or decrease the level of the toxin in the cells ofAphanizomenon. Growth in N-free medium (i.e. N₂ fixation) showed that the cells contained more toxin than growth in N-rich medium. Orthophosphate concentration had no effect on toxin levels, although the lowest concentrations limited the growth of all strains studied. The toxic strains tolerated higher temperatures than the non-toxic one, but the non-toxic strain seemed to be more adjustable to high irradiance than the toxic ones. The yields (dry weight) of non-toxic and toxic strains differed significantly in different phosphate concentrations.     

Use of Refrigeration as a Practical means to Preserve Viability of in Vitro-Cultured IDE8 Tick Cells

In vitro cultivation of the IDE8 cell line, derived from embryonic Ixodes scapularis ticks, constitutes an important system for the study of tick-borne pathogens, as these cells support growth of rickettsial species which are not normally transmitted by this tick. However, since cryopreservation of IDE8 cells is not always successful, there is a need to develop alternative ways to preserve these cells. In the present study, a suspension of IDE8 cells in culture medium was kept under refrigeration at 4°C for up to 60 days. Every 15 days, the suspension was mixed and aliquots were re-cultured in 2-ml tubes, under standardized conditions. In addition, three techniques for cryopreservation, using two different cryoprotectants (DMSO and glycerol), were evaluated. Medium changes were carried out every week and subculturing every 2 weeks. The development of cultures and their respective subcultures, after returning to standard culture temperature, was evaluated by percentage viability and by cellular morphology evaluated in Giemsa-stained cytocentrifuge smears. All cultures and subcultures appeared healthy, showing growth rates comparable to cultures that had not been kept under refrigeration. The results demonstrated that storage under refrigeration at 4°C is an efficient method for preservation of IDE8 cells for up to 60 days and that refrigeration may be preferable to cryopreservation for short-term preservation of IDE8 cells.