Invalidity of the Concept of Slow Growth and Alkali Production in Cowpea Rhizobia

A total of 103 rhizobial strains representing the cowpea miscellany and Rhizobium japonicum were studied with regard to growth rate, glucose metabolic pathways, and pH change in culture medium. Doubling times ranged from 1.4 ± 0.04 to 44.1 ± 5.2 h; although two populations of “fast-growing” and “slow-growing” rhizobia were noted, they overlapped and were not distinctly separated. Twenty-four strains which had doubling times of less than 8 h all showed NADP-linked 6-phosphogluconate dehydrogenase (6-PGD) activity, whereas only one slow-growing strain (doubling time, 10.8 ± 0.9 h) of all those tested showed 6-PGD activity. Doubling times among fast growers could not be explained solely by the presence or absence of 6-PGD activity (r² = 0.14) because the tricarboxylic acid cycle and the Emden-Meyerhoff-Parnas pathway were operative in both 6-PGD-positive and 6-PGD-negative strains. Growth rate and pH change were unrelated to each other. Fast- or slow-growing strains were not associated with any particular legume species or group of species from which they were originally isolated, with the exception of Stylosanthes spp., all nine isolates of which were slow growers. We conclude that 6-PGD activity is a more distinctive characteristic among physiologically different groups of rhizobia than doubling times and that characterization of the cowpea rhizobia as slow-growing alkali producers is an invalid concept.     

Measurement of growth of bifidobacteria on inulofructosaccharides

A simple method for continuous measurement of growth of bifidobacteria is described. An inoculated broth in a small tube was evacuated at 100mmHg through a two-way stopcock and incubated at 37°C. Growth was periodically measured by inserting the tube into a spectrophotometer. Doubling times of three bifidobacterium strains in broth containing various inulofructosaccharide preparations were compared. Inulofructosacchandes from the Jerusalem artichoke tubers supported good growth of these bacteria.     

Rapid microbial growth in a pH auxostat

Summary Feeding nutrient to meet demand dilutes slow-growing organisms from continuous culture and greatly favors rapid growth. Doubling times of roughly 10 minutes have been verified in a pH auxostat by viable cell counts and by direct counting with a Petroff-Hausser chamber. Effects of wall attachment were negligible because a fresh reactor was substituted frequently before wall growth could become established.     

Early Holocene population expansion of some rainforest trees at Lake Barrine basin,Queensland

Pollen influx data derived from a sediment core from a small crater lake were related to two population growth models. Doubling times for some tropical rainforest tree taxa, estimated from the models, are 63–165 years for angiosperms and 199–355 years for conifers. These values are generally higher than those for temperate forest trees in the Northern Hemisphere. The results show that fossil pollen data can provide useful information on tree population ecology in tropical areas.     

The effect of temperature on the growth of non-proteolytic type B Clostridium botulinum

The effect of temperature between 4 and 35°C on the growth rate of a non-proteolytic type B strain of Clostridium botulinum was examined. Growth was in culture media at pH 6.7 and was measured by viable counts using the Most Probable Number (MPN) method. Doubling times were derived from the curve fitting model of Baranyi et al. (1992) and ranged from 42.3 h at 3.9°C to 22 min at 35°C.     

Shoot doubling time: A quantitative parameter for characterizing shoot cultures in vitro

Shoot doubling time is proposed as a suitable parameter for characterizing in vitro propagation rates in shoot cultures. Doubling times were estimated for cultures of black currant and apple; for black currant, doubling times ranging from 14 to 115 days were obtained, depending on the concentration of benzyladenine in the culture medium. These doubling times, which were constant for each culture, were maintained for periods of up to 76 days and could apparently be sustained indefinitely. For one subculture period only, a shoot doubling time as short as 5.6 days was obtained for black currant at high cytokinin concentration (3 × 10⁻⁵ M), but this rate could not be sustained. Shoot doubling time is a convenient parameter for use in optimizing proliferation rates in shoot cultures; its use may also facilitate investigations into the mechanisms of processes underlying shoot proliferation in vitro.     

Sodium butyrate-induced alteration of growth properties and glycogen levels in cultured human colon carcinoma cells

Summary The effect of sodium butyrate on three cultured human colon carcinoma cell lines was studied. Exposure to butyrate caused morphological changes and resulted in the alteration of several growth properties. Doubling times of treated cells were increased five-fold and saturation densities and cloning efficiencies were decreased. compared to untreated cells. Histochemical studies using the periodic acid-Schiff reaction in conjunction with diastase digestion showed that butyrate induced increased glycogen levels in all three cell lines. This increase was confirmed by biochemical techniques. These effects of butyrate were reversed when treated cells were subsequently grown in the absence of butyrate. These changes are consistent with findings from several laboratories that butyrate can induce, phenotypic changes in cultured tumour cells.     

NUTRITIONAL STUDIES ON THE CHLOROMONADOPHYCEAE: VACUOLARIA VIRESCENS AND GONYOSTOMUM SEMEN12

Unialgal, nonaxenic cultures of Vacuolaria virestcens Cienkowsky were grown in soil-water medium, in simple salt solutions with added peat and soil extract, and in synthetic medium. Doubling times in different media were determined: maximum growth rate (doubling time = 46.0 hr) was obtained in continuous light (210 ft-c) at 24 ± I C. Gonyostomum semen (Ehrenberg) Diesing was isolated from a natural population and grown in soil-water medium and a synthetic medium. Other techniques that have been used for culturing chloromonadophycean algae are reviewed.     

Oxidation of Elemental Sulfur by Sulfolobus acidocaldarius

Oxidation of elemental sulfur by Sulfolobus acidocaldarius, an autotroph which grows at high temperatures and low pH, was examined by use of (35)S-labeled elemental sulfur. When cultured at pH 3.2 and 70 C, S. acidocaldarius oxidized elemental sulfur essentially quantitatively to sulfuric acid. Oxidation rate paralleled growth rate and decrease in pH of the culture medium. Elemental sulfur was not oxidized under these conditions if the culture was poisoned with formaldehyde. During the growth phase, the proportion of cells attached to the sulfur crystals increased progressively, and in the later phases of growth over 10 times more cells were attached to sulfur than were free. Doubling times for eight strains growing on elemental sulfur varied from 37 to 55 h. The organism grows much more rapidly on yeast extract than on sulfur. In a medium containing both sulfur and yeast extract, sulfur oxidation was partially inhibited, although growth was excellent.     

Isolation and properties of psychrotrophic and psychrophilic, pectolytic strains of Clostridium

Fourteen strains of pectolytic clostridia have been isolated that were capable of growth at 5–10°C in 7 d; two strains were psychrophiles and failed to grow at 20°C in 14 d and the remainder were psychrotrophs. The bacteria formed pectate lyase enzymes and were capable of degrading potato tissue; they are therefore a potential cause of soft rot of potatoes stored at low temperatures. Doubling times for representative strains were 15–19 h at 10°C and 21–53 h at 5°C. Twelve strains were classified with Group I Clostridium species and two strains with Group II. In the case of one strain the mature spores were not released from the sporangium. Electron microscopy of ultrathin sections of this strain showed the presence of disorganized lamellar structures associated with the spore coat.     

Development and Properties of Streptomycin Resistant Cultures of Erwinia amylovora Derived from English Isolates

Resistance to 1000 p/m, streptomycin was developed in 3 out of 16 virulent strains of Erwinia amylovora by continuous subculturing on increasing concentrations of the drug. Resistance to various antibiotics, including streptomycin, was more readily developed in strains of Erwinia herbicola . Streptomycin resistance carried on an R factor was transferred by conjugation from Escherichia coli to E. amylovora and to E. herbicola . Resistance to streptomycin was associated with attenuation or, in one case, complete loss of virulence. Doubling times of resistant cultures were greater than those of the parent culture both in shaken broth culture and (with the two attenuated cultures) in apple seedlings. The avirulent culture appeared to persist longer in vivo in the presence of the virulent culture than alone.     

Inhibition of Escherichia coli growth by cyclic adenosine 3', 5'-monophosphate

Cyclic AMP inhibits growth rate of E. coli Hfr 3000. Doubling times in glucose minimal medium increased from 60 to about 90 minutes with the addition of 5 mM cAMP. This effect is specific since it was not observed when the cyclic nucleotide was replaced by 5′ AMP, ADP, ATP or adenosine. Half maximal inhibition was obtained with 1 to 3 mM cyclic AMP. This inhibition occurs only with those carbon sources which are known to decrease intracellular cyclic AMP levels, i.e. glucose and pyruvate. No inhibition was observed with succinate, malate or glycerol.     

Transformation and characterization of mutant human fibroblasts defective in peroxisome assembly.

Human skin fibroblasts deficient in peroxisome biogenesis were transformed by transfecting SV40 ori- DNA with the use of an electroporator, and the biochemical, immunocytochemical, and cytogenetic properties of the transformants were analyzed. Cells (1 x 10(6)) from a patient with Zellweger syndrome and one with neonatal adrenoleukodystrophy were suspended with 2 micrograms of SV40 ori- DNA in PBS; then a high-voltage pulse (2000 V, 30 microseconds) was generated two times. Several colonies expressing large T-antigen were picked up 4 weeks after transfection. Doubling time of the transformants was about half of that and the saturation density was 5 to 10 times greater than that of the parental cells. Biochemical abnormalities including defective lignoceric acid oxidation, dihydroxyacetone phosphate acyltransferase deficiency, and disturbed biosynthesis of peroxisomal beta-oxidation enzymes were preserved in the transformants. Peroxisomes were defective in all colonies, as determined by immunofluorescence staining using anti-catalase IgG. Cell fusion studies confirmed that the transformants belong to the same complementation groups as those of the parental cells. These transformed mutant cell lines are expected to be useful tools for investigating the pathogenesis of inherited diseases related to defects in peroxisome biogenesis.     

Factors influencing cell shape in the mutans group of streptococci.

Electron and light microscopic and growth studies of representatives of the diverse species of mutans streptococci revealed the cells to be either bacillary or coccoid in shape. Some strains changed from bacillary to coccoid if the HCO3-/K+ ratio of the media was increased and from coccoid to bacillary if the ratio was decreased. Doubling times of rods and cocci were the same despite an HCO3-/K+ ratio change between 0.008 and 2.84. For strain 10449S, no tested anions or cations substituted for HCO3- or K+ to produce this effect, except for B4O7(2-). Strain 10449S grown at a high B4O7(2-)/K+ ratio became ellipsoid, and this phenomenon was associated with slower doubling times. Up to three incomplete septa could be observed in one rod, but no more than one incomplete septum could be observed in either ellipsoid or spherical cells. Interseptal distances were greatest in rods, shorter in spheres, and shortest in ellipses. All of the above differences were statistically significant (P less than 0.001).     

Phase-Shifting of Cycles in Level of Serotonin N-Acetyltransferase Activity and Cyclic GMP Content of Cultured Chick Pineal Glands by Methotrexate

Methotrexate at 1 microM stimulated increase of serotonin N-acetyltransferase (NAT) activity in chick pineal glands cultured under each of three conditions of illumination. The peak of the circadian rhythm in NAT activity and the "spike" in content of cyclic GMP were both advanced in pineal glands cultured in the dark from midphotoperiod. In contrast, the time of peak NAT activity in glands cultured in the dark from late photoperiod was unaffected. In addition, methotrexate did not affect times of reaching maximum NAT activities in glands cultured from midphotoperiod in the light or under diurnal illumination. Doubling the concentration of methotrexate also eliminated the lag phase in increase of NAT activity in glands cultured in the dark. However, at a concentration of 5 microM methotrexate the curve depicting increase of NAT activity was biphasic, and neither time nor level of peak NAT activity differed from those of control glands. Results of attempts to demonstrate persistent effects of exposure to methotrexate were inconclusive.     

Characterization of multiple chlorobenzoic acid-degrading organisms from pristine and contaminated systems: mineralization of 2,4-dichlorobenzoic acid

Multiple bacterial strains with CBA metabolic properties were isolated using a simple selective strategy. Phylogenetic analysis of the 16S rRNA gene sequences grouped them into two main clusters consisting of four bacterial phyla and belonging to 17 genera. Whereas growth was more frequent with 2-CBA (～68%), 50% grew on 4-CBA and ～7% utilized 3-CBA. One third of the strains exhibited 2,4-dichlorobenzoic acid (2,4-diCBA) catabolic function and were mainly representatives of α-, β- and γ-Proteobacteria. In batch experiments, growth was concomitant with substrate disappearance and near-stoichiometric release of chloride. Doubling times for 2,4-diCBA degradation doubled those determined for mono-substituted CBAs. Out of the six 2,4-diCBA degraders submitted for enzyme assays, significant induction of catechol 1,2-dioxygenase types I and II activities in cell-free extracts were found in four while protocatechuate 3,4-dioxygenase activity was detected in the remaining two. Activities in CBA-grown cells were 20 orders-of-magnitude higher than those grown on benzoic acid.     

Biochemical characteristics of C-6 glial cells

C-6 glial cells were studied in culture with respect to morphological and biochemical changes under several experimental conditions. Doubling time was 33 hr for cells plated at either 0.5 or 1.0×10⁶ cells per flask. Markedly reduced cell growth and astrocyte-like appearance were observed following dibutyryl cyclic AMP (DBcAMP) treatment. An inverse relationship between cell density and DNA, RNA, and protein content per cell was observed. AChE and BuChE activities were also inversely related to cell density, and treatment with DBcAMP increased enzyme activity, but did not alter the cell density relationship. Uptake of³H-norepinephrine also decreased with increasing cell density. In DBcAMP-treated cells,³H-NE uptake was markedly lower than in nontreated controls, and cortisol treatment decreased the uptake of³H-NE in DBcAMP-treated cells further still. We interpreted the foregoing changes to indicate that cellular activity is cell density-dependent.     

Trypanosoma theileri: In vitro cultivation in tsetse fly and vertebrate cell culture systems

Epimastigote forms of Trypanosoma theileri were grown at 25°C in insect cell culture media and in Glossina tissue cultures for more than 6 months. Doubling times of 10–14 h during exponential growth were observed. In cell cultures which had been derived from pupal tsetse flies growth rates were higher than in cell free media; in a larval cell line, however, growth of T. theileri was inhibited. Ecdysteroids and juvenile hormone I reduced multiplication of T. theileri in cell free media. When T. theileri was incubated in different sera only fetal calf serum (FCS) supported growth. Epimastigote forms transformed into trypomastigote bloodforms when cultured at 37°C in FCS, vertebrate cell cultures, and Eagle's medium, but not in insect media or Glossina cell cultures. Oxygen uptake of epimastigotes could be inhibited by rotenone antimycin A and cyanide; trypomastigotes were not affected by these inhibitors.     

闽南-台湾浅滩近岸上升流区浮游植物碳同化速率的研究

本文采用~(14)C同位素示踪法,测定了浮游植物光合作用速率,结合浮游植物细胞含碳量,计算了不同季节、不同深度下浮游植物碳同化速率常数,讨论了不同环境条件对浮游植物碳同化速率常数的影响。结果表明,闽南-台湾浅滩近岸上升流的形成,是该海域高生产力的主要原因,上升流期间浮游植物复制时间要比非上升流期间浮游植物复制时间缩短1.8倍。同时还表明,温度,光照强度、营养盐是控制浮游植物生长的主要因子。上升流期间营养盐始终保持在较高的水平。是上升流区具有较大的浮游植物碳同化速率常数(3.3d~(-1))的原因之一。适宜该海域浮游植物生长的光照强度在3 000—15 000lx之间,温度的影响可用Goldman和Carpenter模式近似表示。     

Effects of nutrient supply (NPK) on spring wheat response to elevated atmosperic CO2

The effects of increased atmospheric CO₂ on crop growth and dry matter allocation may change if nutrient supply becomes insufficient for maximal growth. Increased atmospheric CO₂ may also cause changes in minimum nutrient concentration in plant tissue and hence in the nutrient use efficiency or yield-nutrient uptake ratios of crops. To study these effects for spring wheat, pot experiments have been carried out in two glass houses at ambient and doubled CO₂ concentration. Wheat plants were grown at different supplies of N, P or K. Doubling of ambient CO₂ resulted in a large increase in total biomass (+70%) and grain yield when the nutrient supply was optimum. With strong N and K limitation this CO₂ effect was about halved and with strong P limitation it became almost nil. Doubling of ambient CO₂ resulted in a 10% lower minimum N concentration in plant tissue and in no change in the minimum P concentration.