Gypsy moth cell lines divergent in viral susceptibility

Summary A series of cell lines unique in insect virus susceptibility pattern have been isolated from the ovaries of the gypsy moth (Lymantria dispar: Lepidoptera: Lymantriidae) on a synthetic medium with mammalian and avian serum supplementation. Growth curves showed the poorest growth occurring on peptone-based media with somewhat better growth on amino-acid-based media. The best growth was obtained with combined media. Serological study distinguished the present cell lines from one another and from cell lines derived from other insect species grown routinely in the same laboratory. Baculovirus susceptibility among the new lines varied from no response to a specific complete replication response upon challenge by the homologous (gypsy moth) nuclear polyhedrosis virus. This research was funded in part through a reimbursable agreement with the U.S. Forest Service.     

Physical conditions of propagation media and their influence on the rooting of cuttings

Summary The air content in three types of propagation media, Jiffy-7 and Jiffy-9 which are Sphagnum peat and Grodan which is rockwool, were investigated when they were held at moisture tensions of 0,6 and 12 cm measured from the base of the media. At 0 cm tension the air content (vol. %) was highest in Jiffy-9 and lowest in Jiffy-7. At 12 cm tension the air content was higher in Grodan than in Jiffy-9 and Jiffy-7. Oxygen diffusion coefficients (ODC) and oxygen diffusion rates (ODR) were measured at the different air contents. At air contents below 20 vol. % ODC was about the same for Jiffy-9 and Grodan but at air contents above 20 vol.% it was larger for Jiffy-9 than for Grodan. The oxygen diffusion rate was measured at 0, 4 and 8 cm moisture tension. At all tensions it was approximately 20% higher in Jiffy-9 than in Grodan and Jiffy-7. The ODR in Jiffy-7 and Grodan were affected equally at the same tension, although Grodan contained more air. Report no 253     

An examination and correction of plant tissue culture basal medium formulations

The inorganic formulations of fourteen common plant tissue culture basal media were examined from the primary literature. Inaccuracies and errors were found for molecular formulae, chemical hydrations, and molar equivalences for iron/EDTA complexation. A comparison with published basal medium formulations from six commercial suppliers uncovered additional inaccuracies, modifications, and errors, thereby emphasizing the need for investigators to examine and describe medium formulations precisely in future publications.     

Impact of the field slug Deroceras reticulatum on establishment of ryegrass and white clover in mixed swards

The relationship between numbers of Deroceras reticulatum and the establishment of ryegrass cv. Melle and white clover cv. Milkanova was studied on a mini-plot experiment. Assessments 12 and 33 days after sowing showed that Deroceras reticulatum had a proportionally greater impact on survival and growth of clover than on ryegrass. Slugs had no apparent effect on numbers or growth of three species of broad-leaved weeds (Senecio vulgaris, Cardamine hirsuta and Epilobium sp.) which occurred in the field experiment. In pot experiments where seedlings of ryegrass together with one of three clover cultivars, with or without chickweed (Stellaria media), were exposed to Deroceras reticulatum, seedlings of white clover cv. Aran were less affected by slug grazing than cvs S184 or Milkanova. The presence of different clover cultivars did not significantly influence the impact of slugs on growth of ryegrass or chickweed, nor did the presence or absence of chickweed influence the effect of slugs on ryegrass and clover seedlings. Slugs had less impact on the growth of chickweed than on ryegrass or clover. Thus slug grazing would make clover less able to compete with chickweed as well as the other weeds found in the mini-plot experiment.     

Serum-free medium for fermentor cultures of hybridomas

Hybridomas lend themselves particularly well to large scale cultivation techniques since they grow as single cells in suspension without requiring attachment to a substrate. Furthermore, many cell strains have been adapted to grow in serum-free (SF) media to a similar cell density and antibody production as in serum containing media. This review will concern itself mainly with the cultivation of hybridomas in SF-media in bioreactors of various types with the ultimate goal of producing large quantities of monoclonal antibodies (mAb).     

Method for Lyophilizing Brain Proteolipid Preparation That Increases Subsequent Solubilization by Detergent

Abstract: A frozen mixture of solubilized brain proteolipid proteins in chloroform-methanol is not sublimable in a vacuum. However, when 7 to 10 volumes of benzene were added to a chloroform-methanol solution containing 5 mg of proteolipid protein per ml, the proteolipid proteins remained in solution for a while and the frozen mixture was easily sublimated at 2 mm Hg. Before the addition of benzene, higher concentrations of protein required the acidification of the medium to avoid precipitation of proteolipid proteins. In contrast to what happens when proteolipid proteins are obtained by the evaporation of the organic mixture at room temperature, the protein obtained by lyophilization was soluble in aqueous solutions of ionic and nonionic detergents. Sodium dodecyl sulfate at 0.6 to 0.7% concentration completely solubilized the proteolipid protein obtained by lyophilization. With the nonionic detergents Lubrol WX and Triton X-100, a solubilization between 50 and 65% was achieved. Sodium deoxycholate was practically ineffective. Triton X-100 showed selectivity in solubilizing certain proteins. The role of lipids in the solubilization of proteolipid proteins with detergents is discussed.     

Benomyl and mancozeb as fungicides in soil studies of Azotobacter and Beijerinckia

Summary Routine media containing 0.005% each of benomyl and mancozeb for the enumeration and isolation of Azotobacter and Beijerinckia in soil studies are described. The incorporation of these fungicides in routine media was found to be more effective in inhibiting fungal contaminants than the use of cycloheximide.     

In vitro development of theDrosophila chorion in a chemically defined organ culture medium

Summary TheDrosophila chorion is produced normally in isolated follicles in Robb's chemically defined culture medium. The complex architecture of the shell developed in vitro from follicles as young as early stage 10 is completely normal morphologically. In addition, the time required for in vitro development closely approximates that observed for in vivo development. Comparisons of insect culture media developed by Robb, Grace, Schneider, and Echalier show large variations in their ability to supportDrosophila chorion development.     

Leishmania tarentolae: streptomycin and chloramphenicol resistance of promastigotes.

Stable mutant strains of Leishmania tarentolae promastigotes resistant to chloramphenicol (CAP) were isolated by replica-plating techniques. In addition, cell lines stress-adapted to streptomycin and to high culture temperature (33 C) were obtained. Drug resistance was influenced by temperature, culture media, and plating technique. Inhibition of amino acid incorporation into protein occurs in CAP-resistant cells when exposed to 600 μg CAP/ml but this inhibition was 50–80% lower than that found in wild type sensitive cells. The primary site of CAP action appears to be inhibition of protein synthesis. CAP also adversely affected proline oxidation.     

Design of an efficient fermentation process for the production of Metarhizium acridum blastospores

Using a sequential approach, we described efficient blastospore production in a stirred tank bioreactor (3?L capacity). We used the response surface methodology to optimise the media ingredients and fermentation parameters to obtain the maximum production of blastospores by a locally collected isolate of Metarhizium acridum (Ascomycota: Hypocreales). The results showed that a liquid culture medium supplemented with monopotassium phosphate (15.17?g/L), corn steep liquor (69.25?g/L), and casamino acids (80.68?g/L) in a stirred tank bioreactor under operating conditions constant at 635?rpm, a temperature of 26°C, and pH 3.3 produced 1.25?×?10⁸?blastospores (bls)/ml, with 93% viability after 120?h of fermentation. This bioreactor yield compares favourably with the yields obtained by shake flask production and confirms the suitability of the media and production parameters for the potential scale-up fermentation production of M. acridum.