Radial growth rate of soil Micromycetes colonies under different conditions of nitrogen nutrition

The radial growth rate of soil micromycetes colonies as a function of mineral nitrogen concentrations in the medium is expressed by a bell-shaped curve. Low nitrogen concentrations are growth-limiting whereas its high concentrations inhibit the growth. Soil micromycetes differ in the absolute values of growth rates and in the ranges of tolerance.     

Primary culture and cryopreservation of mouse astrocytes under serumfree conditions

The methods of primary culture and cryopreservation of mouse astrocytes under serum-free conditions were examined. Cerebra from newborn C3H/He mice were employed as the source of astrocytes. The cultured cells were able to grow in a serum-free, chemically defined medium containing transferrin, hydrocortisone, biotin, sodium selenite, insulin, fibroblast growth factor and epidermal growth factor. After the culture was maintained in the medium for 3 weeks, purity was assessed using immunofluorescence staining. The great majority of the cells (>98%) contained glial fibrillary acidic protein and S-100 protein which are cell markers of astrocytes. To cryopreserve the enriched astrocytes under serum-free conditions, various cryoprotectants were examined. The combination of 10% dimethylsulfoxide and 0.1% methylcellulose gave the highest survival rate. These methods of primary culture and cryopreservation will be useful in physiological and biochemical studies which require mouse astrocytes.     

Micromycetes metabolites--inhibitors of growth and sterol biosynthesis in yeasts]

Antifungal activity of micelial fungus metabolites (of genera Aspergillus, Penicillium, Fusarium, Stachybotris, Cladosporium, Alternaria, Gliocladium, Paecilomyces, Trichoderma etc.) was determined. It was shown that antifungal activity of some micromycetes is due to the formation of substances inhibiting sterols biosynthesis in eucaryote cells. Inhibitors of enzymes of sterols biosynthesis were isolated and their activity was investigated. It was shown, that isolated fungus inhibitors of sterols biosynthesis inhibited the growth of test-organism Rhodotorula rubra and decreased ergosterin level in yeast cells. The qualitative content of yeast cell sterols was not changed in the presence of fungus inhibitors.     

Intensity of the epidemic process of whooping cough under conditions of mass immunization over a period of many years

On the basis of analysis of whooping cough incidence in 1959 to 1975 in Moscow the authors present characteristics of the epidemic process under conditions of planned many-year immunization of children against this infection. Whooping cough morbidity proved to undergo significant changes-from sharp falls the first 8 years of immunization to a relative stabilization with a gradual reduction the last 5 years. Marked seasonal and periodic elevations persist against the background of reduction of morbidity. Preschool- and schoolchildren are equally involved in the epidemic process; the most frequently involved are nonvaccinated children aged 1 year and 4 to 10 years, which lost postvaccinal immunity. Further reduction of the severity of whooping cough and increase in the number of unimanifested forms of the infection was noted. This was apparently connected not only with immunization, but also with the reduction of the virulence of H. pertusis detected in studying the cultures isolated in Moscow from 1967 to 1974. The intensity indices of the epidemic process in whooping cough pointed to the necessity of using a more effective vaccine for the protection of children from whooping cough, despite the noted reduction of morbidity, and diminished severity of the course of the disease, and of the causative agent virulence.     

Methods for detecting drug-resistant staphylococci in the human body under natural conditions]

Two methods were compared for determination of drug resistant staphylococci on the nasal mucosa of patients, i. e. the routine method for determination of staphylococcal sensitivity to antibiotics and the method of direct plating out of the starting material onto agarized media containing antibiotics. Staphylococci resistant to streptomycin, tetracycline, chloramphenicol, erythromycin, monomycin, axacillin and less frequent to penicillin were found more often with the 2nd method. A method of proportions was developed for testing sensitivity of staphylococci in purulent inflammatory foci of the patients. It provided characterization of the staphylococcal population from the foci by the number of the antibiotic resistant microbial cells.     

Drought-tolerant endophytic actinobacteria promote growth of wheat (Triticum aestivum) under water stress conditions

Drought tolerant endophytic actinobacteria Streptomyces coelicolor DE07, S. olivaceus DE10 and Streptomyces geysiriensis DE27 were isolated from cultivated plants of arid and drought affected regions of Rajasthan, India. These isolates exhibited plant growth promotion traits and intrinsic water stress tolerance from ?0.05 to ?0.73 MPa. Maximum auxin production was observed in majority of actinobacterial cultures in the logarithmic to stationary phase of growth. Significant enhancement of wheat seedling vigour was recorded by the inoculation of these endophytic actinobacteria. S. olivaceus DE10 recorded maximum accumulation of indole 3-acetic acid (84.34?μg?mg^?1 protein). Culture and cell-free extract of the endophytes was applied on to wheat seeds to assess the effect on growth in water-stressed soil. Maximum yield was recorded with the inoculation of S. olivaceus DE10 culture (492.77?kg?ha^?1) and cell-free extract (262.31?kg?ha^?1). Co-inoculation of S. olivaceus DE10?+?S. geysiriensis DE27 recorded highest yield of 550.09?kg?ha^?1 while their cell-free extract yielded 524.92?kg?ha^?1. Overall, wheat seeds treated with cultures showed better plant growth and yield in comparison to control. Direct coating of cultures on seeds yielded better performance than cell-free extract coated on seeds and co-inoculation of cultures or cell-free extract proved better than single culture inoculations. Production of phytohormones, plant growth promotion traits combined with water stress tolerance potential in these endophytic actinobacteria played a cumulative synergistic role that supported enhanced plant growth promotion of wheat in the stressed soil.     

Survival of Micromycetes and Actinobacteria under Conditions of Long-Term Natural Cryopreservation

Almost all of the investigated samples of the Arctic and Antarctic permafrost sediments of different genesis with ages from 5–10 thousand to 2–3 million years were found to contain viable micromycete and bacterial cells. The maximum amounts of viable cells of fungi (up to 10⁴CFU/g air-dried sample) and bacteria (up to 10⁷–10⁹CFU/g air-dried sample) were present in fine peaty sediment samples taken from different depths. The identified micromycetes belonged to more than 20 genera of the divisions Basidiomycota, Ascomycota, and Zygomycota, and some represented mitosporic fungi. Thawing the samples at 35 and 52°C allowed the number of detected fungal genera to be increased by more than 30%. Aerobic heterotrophic prokaryotes were dominated by coryneform, nocardioform, and spore-forming microorganisms of the order Actinomycetales.Analysis of the isolated fungi and actinomycetes showed that most of them originated from the microbial communities of ancient terrestrial biocenoses.     

Marine actinobacteria: new opportunities for natural product search and discovery

It is widely accepted that new drugs, especially antibiotics, are urgently required, and that the most propitious source remains natural products. We argue that in exploring new sources of bioactive natural products the marine environment warrants particular attention, in view of the remarkable diversity of microorganisms and metabolic products. Recent reports of new chemical entities and first-in-class drug candidates, and confirmation of indigenous marine actinobacteria, make exciting discoveries even more likely given the unrivalled capacity of this class of bacteria to produce exploitable natural products.     

Degradation of natural and synthetic polyesters under anaerobic conditions

Often, degradability under anaerobic conditions is desirable for plastics claimed to be biodegradable, e.g. in anaerobic biowaste treatment plants, landfills and in natural anaerobic sediments. The biodegradation of the natural polyesters poly(beta-hydroxybutyrate) (PHB), poly(beta-hydroxybutyrate-co-11.6%-beta-hydroxyvalerate) (PHBV) and the synthetic polyester poly(epsilon-caprolactone) (PCL) was studied in two anaerobic sludges and individual polyester degrading anaerobic strains were isolated, characterized and used for degradation experiments under controlled laboratory conditions. Incubation of PHB and PHBV films in two anaerobic sludges exhibited significant degradation in a time scale of 6-10 weeks monitored by weight loss and biogas formation. In contrast to aerobic conditions, PHB was degraded anaerobically more rapidly than the copolyester PHBV, when tested with either mixed cultures or a single strained isolate. PCL tends to degrade slower than the natural polyesters PHB and PHBV. Four PHB and PCL degrading isolates were taxonomically identified and are obviously new species belonging to the genus Clostridium group I. The depolymerizing enzyme systems of PHB and PCL degrading isolates are supposed to be different. Using one isolated strain in an optimized laboratory degradation test with PHB powder, the degradation time was drastically reduced compared to the degradation in sludges (2 days vs. 6-10 weeks).     

自然条件下福寿螺繁殖特性

为了弄清福寿螺在自然条件下的繁殖特性,在湖南省资兴市水稻田内进行了为期3年的人工养殖试验.结果表明:福寿螺在湘南自然条件下1年可发生不完全3代.第1代、第2代和第3代(越冬代)雌螺的平均性成熟期分别为59.3、45.4和213.0 d.雄螺的平均性成熟期比雌螺早4.3d.福寿螺的自然性比(♀∶♂)为1.54:1.每次交配持续时间平均为19.2h,但并非每次交配后都会产卵.田间福寿螺月产卵块数与月平均气温呈显著正相关(r=0.756).福寿螺卵块的平均孵化期(d)与日平均气温(℃)呈显著负相关(r=-0.980),其线性回归方程为y=-0.726x+23.064.福寿螺卵块的平均孵化期为20.7d,平均孵化率为44.1％.雌螺与雄螺的平均寿命分别为2.40和1.98 a.平均每头雌螺一生可产卵13764粒,可繁殖幼螺6070头.     

Leukocytolysis and phagocytosis under hyperosmotic conditions]

PAN and leukocytolysis were measured in dog blood placed in hypo- and hypertonic solutions of glycerin and glucose (1:10). It was found that leukocytolytic activity of the substances depends on equimolar concentrations of nonelectrolytes. It is more pronounced in hypo- than in hypertonic conditions. Glycerin is 3-5-times more active than glucose. In concentrations 2.0 M for glycerin and 0.5 M for glucose leukocytolysis is less active. Mononuclears are more resistant than neutrophils in strong solutions. PAN increased by 11-16% in hypotonic solutions. Strong glycerin solutions reduce PAN less than glucose which stimulates the digestive activity. The results can be of use in specific leukolysis tests to diagnose drug allergy.     

Modeling Daphnia population dynamics and demography under natural conditions

Various approaches to modeling the population dynamics and demography of Daphnia have been published. These methods range from the simple egg-ratio method, to mathematically complex models based on partial differential equations and numerically complex individual-based Daphnia population models. The usefulness of these models in unraveling the population dynamics and demography of Daphnia under natural conditions is discussed. Next to this, an extended version of an existing individual-based Daphnia model is documented (Cladosim) and its application to a typical field data set collected in 1995 in Lake Volkerak is shown. To answer the question which factor was limiting Daphnia numbers during the course of the season food level and temperature in the model were varied and results were compared with those obtained for the observed food level and temperature. These analysis showed that in April temperature was limiting while during May–July and September–October food was limiting. In August neither temperature nor food was limiting. Analysis with a set of size-selective mortality scenarios showed that on average the Daphnia population in Lake Volkerak experienced a mild positive size-selective mortality during the year that was analyzed. Birth rates derived with the detailed individual-based model were compared with those derived with the much simpler egg-ratio method. For the conditions as observed in Lake Volkerak in 1995, both methods gave very comparable results, despite sampling intervals of up to four weeks. The same holds under the environmental scenarios. Using the size-selective mortality scenarios it could be shown, however, that under strong mortality of the smaller daphnids, the egg-ratio method severely underestimates the birth rate. The vices and virtues of the new model and potential extensions are discussed.     

Hydrogen peroxide concentrations in leaves under natural conditions

While H2O2 has been implicated in numerous plant environmental responses, normal levels and variabilities are poorly established, and estimates of leaf tissue concentrations span more than three orders of magnitude, even in a single species under similar conditions. Here, leaf tissue H2O2 contents under natural conditions are reported after determining (i) that H2O2 in extracts was stable with time, (ii) that H2O2 added to the extract was recovered quantitatively, and (iii) that the H2O2 calibration curve was unaffected (or quantifiably affected) by the extract. The broad applicability of the protocol and variability in leaf concentrations were demonstrated using tissue collected from several habitats in association with three, more extensive, experiments. The first involved nychthemeral studies of the mangrove, Rhizophora mangle L. Lowest H2O2 levels occurred in the early morning and near sunset, with higher levels both at midday and at night. Second, using five temperate species in Spring, concentrations were compared on a warm, sunny day and a cool, cloudy day. Higher concentrations were found on the warm day for Aesculus glabra Willd., Glechoma hederacea L., Plantago major L., and Viola soraria Willd., while there were no differences in Quercus macrocarpa Michx. Finally, the effects of elevated CO2 and ozone were examined in soybean, Glycine max L. Pioneer 93B15 under Free Air gas Concentration Enrichment (FACE) conditions. Both supplements led to elevated H2O2. Overall, mean leaf, midday, and mid-summer H2O2 concentrations ranged from 0.67 micromol (gFW)(-1) in mangrove to 3.6 micromol (gFW)(-1) in A. glabra Willd. Greatest within-species differences were only 2.5-fold in any of the studies.     

Selective phonotaxis of female crickets under natural outdoor conditions

Acoustic mate choice in insects has been extensively studied under laboratory conditions, using different behavioural paradigms. Ideally, however, mate choice designs should reflect natural conditions, including the physical properties of the transmission channel for the signal. Since little is known about the discrimination ability of females between male song variants under natural conditions, we performed phonotaxis experiments with female field crickets (Gryllus bimaculatus) outdoors, using two-choice decisions based on differences in carrier frequency, sound pressure level, and chirp rate. For all three song parameters, minimum differences necessary for a significant preference between two song models were considerably larger outdoors compared to laboratory conditions. A minimum amplitude difference of 5 dB was required for a significant choice in the field, compared to only 1–2 dB reported for lab-based experiments. Due to the tuned receiver system, differences in carrier frequency equal differences in perceived loudness, and the results on choice for differences in carrier frequency corroborate those in amplitude. Similarly, chirp rate differences of 50 chirps/min were required outdoors compared to only 20 chirps/min in the lab. For predictions about patterns of sexual selection, future studies need to consider the different outcomes of mate choice decisions in lab and field trials.     

Arousing sleeping genes: shifts in secondary metabolism of metal tolerant actinobacteria under conditions of heavy metal stress

Numerous microbial habitats are strongly influenced by elevated levels of heavy metals. This type of habitat has developed either due to ore mining and metal processing or by pedogenesis above metal-rich base rocks. Most actinobacteria are soil-borne microbes with a remarkable capability for the synthesis of a broad variety of biologically active secondary metabolites. One major obstacle in identifying secondary metabolites, however, is the known phenomenon of sleeping gene clusters which are present, but silent under standard screening conditions. Here, we proceed to show that sleeping gene clusters can be awakened by the induction in heavy metal stress. Both, a chemical and a biological screening with extracts of supernatant and biomass of 10 strains derived from metal contaminated and non-contaminated environments was carried out to assay the influence of heavy metals on secondary metabolite patterns of metal tolerant actinobacteria. Metabolite patterns of cultures grown in complex and minimal media were compared to nickel (or cadmium) spiked parallels. Extracts of some strains grown in the presence of a metal salt displayed intense antibiosis against Escherichia coli, Mycobacterium smegmatis, Staphylococcus aureus and Candida albicans. Contrarily to the widely held opinion of metals as hindrance in secondary metabolism, metals thus can induce or enhance synthesis of possibly potent and medically relevant metabolites in metal tolerant strains. Hence, re-screening of existing strain libraries as well as identification of new strains from contaminated areas are valid strategies for the detection of new antibiotics in the future.     

Synthesis and cytotoxicity of pyranonaphthoquinone natural product analogues under bioreductive conditions

We have synthesised a focused library of derivatives of natural products containing the pyranonaphthoquinone moiety including the first report of such a scaffold with an appended tetrazole functionality. Examples include kalafungin derivatives as well as analogues of nanaomycin and eleutherin. These compounds were assessed for cytotoxic activation by breast cancer cell lines engineered to express the prototypic human one- and two-electron quinone bioreductive enzymes, NADPH: cytochrome P450 oxidoreductase (POR) and NAD(P)H: quinoneoxidoreductase 1 (NQO1; DT-diaphorase), respectively. Several compounds were observed to be cytotoxic at sub-micromolar level and a pattern of increased aerobic potency was observed in cells over expressing POR. A subset of analogues was assessed under anoxic conditions, where cytotoxicity was reduced, implicating redox cycling as a major mechanism of toxicity. The substrate specificity for reductive enzymes is relevant to the future design of bioreductive prodrugs to treat cancer.