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[Growth substance requirements in heterothallic yeast cultures].

Vitamin heterotrophy was studied among the following new yeast genera: Leucosporidium, Rhodosporidium and Filobasidium. The requirement in growth factors of various crossing types was compared among these genera and ascomycetens yeast cultures Saccharomycopsis lipolytica and Metschnikowia pulcherrima. Various crossing types of Filobasidium capsuligenum were found to differ in their requirement in growth factors. Various crossing types of Rhodosporidium and Saccharomycopsis lipolytica were stimulated by different growth substances. These characteristics can be used as markers while selecting the types of crossing.

Ascomycota

Biochemical changes in Pinus pinea seeds. III. The effect of growth substances and steroidal hormones on nucleic acids.

The effects of exogenous growth factors (indolacetic acid, gibberellic acid and kinetin and steroidal hormones (estrone, estradiol and testosterone on the germination pattern of Pinus pinea seeds were studied. Nucleic acids metabolism during the stages before germination, has also been investigated. Seeds sown in the presence of all these substances, showed a higher growth rate and a higher germination degree than their respective controls; kinetin and estradiol were the most active factors. The level of total nucleic acids was studied in seeds after one day soaking and on the 1rst, 3rd, 5th, and 7th day after sowing. Megagametophytes contained a higher amount of acids when growth factors and steroidal hormones were present, specially after one day soaking. In embryos; after the 3rd day, a similar result was obtained. The RNA fraction increased the most.

DNA

[Deformation of lucerne root hairs caused by the growth substances and culture broth filtrates of Rhizobium meliloti].

Deformation of lucerne root hairs caused by the action of beta-indolylacetic acid (IAA) and alpha-naphthylacetic acid (NAA) differs from deformation induced by Rhizobium meliloti. High concentrations of IAA brought about abnormal deformation of root hairs wherease the action of average concentrations of IAA resulted in corkscrew-like winding and wavy structures. No deformation was observed under the action of low IAA concentrations or the cultural broth of Rhizobium. Infective filaments in root hairs were found only upon infection with pure cultures of Rhizobium.

Acetates

Biochemical changes in Pinus pinea seeds. II. The stimulation is isocitrate lyase and malate dehydrogenase activities by exogenous growth substances.

Treatment of intact pine seeds with plant hormones and steroid substances for several days, after an initial 24 hour immersion in different hormone solutions, resulted in a substantial increase in the specific activity of isocitrate lyase and malate dehydrogenase than the observed in the corresponding control solutions. Fresh weights did not show any increase, because water uptake was reduced in presence of plant hormones (indolacetic acid, kinetine and giberellic acid). However, dry weights were consistently greater than those of the control solutions.

Enzyme Induction

Automated system for analytical microbiology V: calibration lines for antibiotics.

The accuracy of an automated system for the microbiological assay of antibiotics was increased by improvement attendant to connection to an on-line computer. The system was used to investigate the suitability of four forms of interpolation formulas by assaying for chlortetracycline and erythromycin. The calibration lines were prepared as point-to-point straight-line approximations and as cubic equations. Cubic equations through four calibration points were preferred. Since the automated system was a four-channel instrument, a separate response line was prepared for each channel. Combining the four response lines into one could substantially degrade the accuracy and precision of assays. A new general equation relating the response of the test organism to concentrations of active materials was used to account for factors in addition to the antibiotic upon the dose-response line. Some of these factors were: diluents, growth substances, relative proportions of mixed antibiotics, pH and buffer capacities of the sample solution and assay broth, salts, and organic compounds in samples and not in standard solutions. The equation was used to show under what conditions the dose-response lines of mixtures and single-component antibiotics could be the same. It could also account for the nonspecific nature of turbidimetric assays. The equation showed assay biases to be caused not by differences in composition of antibiotics in standards and samples but by differences in other substances affecting growth of the test organism. A new dose-response line applicable to assays using Klebsiella pneumoniae was described.

Anti-Bacterial Agents

[Stability of a number of antibiotics, coccidiostats and growth-stimulating substances during manufacture and storage of (medicated) feed pellets (author's transl)].

The effect of processing and storage on the stability of a number of antibiotics, coccidiostats and growth-stimulating substances, viz. lasalocid, monensin-Na, avoparcin, olaquindox, carbadox, sulfadimidine-Na and lincomycin in (medicated) feeds was studied. Mixtures containing the additives were conditioned by steam at a temperature of 80 degrees C, and pelleted. After each stage in processing and following storage of the pellets for a month, the concentrations of antibiotics, etc., were determined and compared with the concentrations as calculated in the original feed mixtures. The results suggested that none of these substances were very susceptible to the conditions of processing applied. All samples of stored pellets were found to contain at least 90 per cent of the concentrations present in the original feed mixtures, the combined negative effects of processing and storage hardly being beyond the margin of accuracy of the analyses.

Animal Feed

Effects of bleomycin on the epidermal content of growth-regulatory substances (chalones).

Hairless male mice were given 2 mg Bleomycin i.p. on two successive days. At different time intervals from 1 to 10 days after the last Bleomycin injection, groups of animals were killed and water extracts of hemogenized skin were made. These extracts, supposed to contain the epidermal G1 and G2 chalones, were injected into female hairless mice, and their growth inhibitory potency determined by two methods. 5 mg of lyophilized crude skin extract were injected i.p. together with Colcemid, and the animals killed 4 hr later. The number of Colcemid-arrested mitoses was determined, and was considered to be a measure of the G2 inhibitor present in the skin extracts. 10 mg of the same extracts were injected i.p., and these animals also got 3H-TdR i.p. 12 hr later, and were killed after a subsequent 30 min. The epidermal LI was determined, and was considered to be a measure of the epidermal G1 factor in the skin extracts. The results obtained were compared to the effect of Bleomycin alone and to the effects of skin extracts from non-Bleomycin-treated animals. The results show that Bleomycin provoked slight alterations in the growth-inhibitory potency of the G1 chalone, whereas significant effects were seen in the G2 chalone, There was an increased amount of growth-inhibiting factors on days 2 and 3, and on days 8-10. The results are discussed and it is concluded that the most probable hypothesis is that Bleomycin, in addition to its known inhibition by accumulation of cells with high growth inhibitory potency. An initial, additional direct effect of Bleomycin on the chalone system cammot be excluded.

Animals

Selection of a yeast strain with optimal utilization of straw hydrolyzates.

The dry nonseparated straw hydrolyzates prepared by mild acid hydrolysis of milled straw contains 25--30% of reducing substances, mostly saccharides with prevalence of xylose. A strain utilizing the neutralized nonseparated hydrolyzates without any growth substances added was obtained by selection and long-term adaptation of an array of yeast strains. The strain, identified as Candida tropicalis 2838, exhibited high cell-growth rate and considerable yield of protein-rich biomass.

Animal Feed

Morphogenesis in Mucor mucedo: mutations affecting gamone response and organ differentiation.

Mutants of Mucor mucedo minus strain that are affected in their trisporic acid (TA) mediated zygophore formation have been isolated. We have found mutants with cold sensitive (cs), with temperature sensitive (ts) and without zygophore formation as well as mutants with unstable zygophores (Zst-). From the appearence of certain pleiotropic phenotypes we deduce a one-dimensional sequence of states of competence of the mycelium to form different organs. TA appears to be a growth substance for zygophores acting on one transient state of competence. The fact that all isolates with lowered response to TA also have a lowered response to the mating type specific TA-precursor P strongly suggests that P has to be converted into TA before inducing zygophore growth. Furthermore, one mutant with lowered sensitivity to TA exhibits an excess zygophore formation in the presence of high TA-concentrations, while high concentrations of P cause a depressed zygophore formation (Fig. 6). Our interpretation of this behaviour is that P acts as an antagonist to TA in the regulation of zygophore growth.

Genes, Regulator

Plasmid-determined tetracycline resistance in Streptococcus faecalis: evidence for gene amplification during growth in presence of tetracycline.

The tetracycline (TG)-resistant Streptococcus faecalis strain DS-5Cl harbors two plasmids designated alpha and gamma with molecular masses of approximately 6 and 35 million daltons, respectively. TC-sensitive variants were derived by storing cells at 45 degrees for 2-3 weeks. Analysis of covalently closed circular DNA from five such variants (derived independently) revealed that in each variant the alpha-plasmid, which normally sediments at 28 S (supercoiled) in a sucrose density gradient, was replaced by a 22S substance. Growth of DS-5Cl in the presence of 150 mug/ml of TC (minimum inhibitory concentration is 250 mug/ml in liquid broth) for a prolonged period of time (50-60 generations) resulted in the disappearance of 28S DNA and the appearance of a heterogeneous covalently-closed circular DNA sedimenting at about 40-48 S. This phenomenon was accompanied by an increase in the level of bacterial TC-resistance, whereby tells were subsequently grown in the absence of TC for 70-80 generations, the heterogeneous DNA disappeared and a typical 28S alpha-plasmid reappeared. The cells also became less resistant to TC, i.e., the minimum inhibitory concentration returned to 250 mug/ml. These data suggest that bacterial growth in the presence of TC results in a reversible gene amplification with respect to a TC-resistant determinant residing on the alpha-plasmid.

Cell Division