Evaluation of Factors Related to Growth of Rift Valley Fever Virus in Suspended Cell Cultures

The effect of several controlled variables on the peak titer and fold increase of Rift Valley fever virus grown in suspension culture on two variants of Earle's L cell, L-DR and L-MA clone 1-1, was studied. No significant amount of cell-associated virus was found at 24 hr, indicating a release of virus soon after its formation. Mild sonic treatment of the virus produced in serum-free medium increased the infective titer about 10x. This difference was not observed with virus produced in medium supplemented with serum. Peak titer was not affected by medium used during the infection period, by multiplicity of inoculum (MOI), or by initial cell concentration within the test range of 10(4) to 2 x 10(6) cell/ml. Cell strain employed influenced titer, because the L-DR cell did not produce virus efficiently at low MOI and low initial cell concentration. The time of peak titer and fold replication was dependent on MOI and initial cell concentration. Differences in virus propagation in monolayer and suspension systems are discussed.     

Microbiological studies of Indonesian fermented foodstuffs

Microbiological studies were made of certain Indonesian foodstuffs obtained from Malang, Surakarta, and Djakarta.Saccharomyces cerevisiae andCandida solani were isolated from ragi-roti (a baker's yeast preparation). From ragi-tempe and tempe were isolatedRhizopus oryzae, R. arrhizus, R. oligosporus, R. stolonifer, Mucor Rouxii, M. javanicus andTrichosporon pullulans. The microbiological flora of ragi-tape was found to includeCandida parapsilosis, C. melinii, C. lactosa sp. nov.,Hansenula subpelliculosa, H. anomala, H. malanga sp. nov.,Chlamydomucor oryzae andAspergillus oryzae. From ragi-ketjap, used to prepare soysauce, were isolatedRhizopus oligosporus, R. arrhizus, Aspergillus oryzae, andA. flavus, the latter species probably being an accidental contaminant. Two new species are described:Candida lactosa andHansenula malanga, from ragi-tape from Surakarta and Malang, respectively.From a thesis presented to the Graduate School of Vanderbilt University in partial fulfillment of the requirements for the Ph.D. degree, August, 1969.Financial support of Dakimah Dwidjoseputro by the Ford Foundation is grate-fully acknowledged.     

Temperature Response in Animals Infected with Bacillus anthracis

Rats, rabbits, swine, guinea pigs, and monkeys were infected with anthrax spores, and their temperature responses were recorded. These were characteristic for a species and appeared independent of resistance or susceptibility of the species toward establishment of the disease. The rabbit appeared unique in that it not only failed to demonstrate a dose-response relationship over an 8-log dose range, but acted independently producing erratic body temperatures depending on spore dose. This limits the usefulness of the rabbit in studying anthrax pathogenesis, and poses questions regarding published data with the rabbit as the test animal.     

Production of a Pyrrole Antibiotic by a Marine Bacterium

Evidence is presented for the isolation and identification of bacteria able to synthesize an unusual antibiotic containing five bromine atoms per molecule. The identification and taxonomic position of these bacteria was made by use of a computer in conjunction with traditional methods. These microorganisms and closely related strains have been isolated on various occasions from tropical water in the vicinity of Puerto Rico. One bacterium, a pseudomonad, has been given the name Pseudomonas bromoutilis because of its distinctive capability. The antibiotic has been extracted, purified, and obtained in crystal form, and its structure has been determined. Although clinical tests of its properties were not encouraging, it may be of significant value and interest from an ecological standpoint.     

Studies on the Bohr Effect of Sheep Hemoglobin

A simple method is described for the measurement of pH changes in hemoglobin solutions on oxygenation and reduction. Data are presented establishing the absence of a Bohr effect in p-chloromercuribenzoate [PCMB] treated hemoglobin. The influence of a number of sulfhydryl inhibitors on the Bohr effect of a hemoglobin solution is reported and an interpretation based on steric factors in the protein is proposed.     

Studies on the Hemolytic Properties of Protamine

The basic protein protamine causes a rapid hemolysis when incubated with the red blood cells of many mammalian species. The age of the cells does not affect the process. Neutralization of the active side groups of the protamine molecule with formalinization demonstrates that a specific degree of charge is necessary for hemolysis, as more than 30 per cent of the guanidine groups must remain unreacted to maintain activity. Unlike the hemolysis induced by the synthetic polypeptides polylysine and polyhomoarginine, protamine hemolysis is temperature-dependent. Whole lipoprotein material derived from red blood cell membranes inhibits protamine hemolysis to a greater extent than do the membranes themselves, serum, serum protein fractions, or cholesterol. The phosphatide and protein moieties derived from the membranes are quite avid in inhibiting protamine hemolysis. A probable explanation is that intracellular aggregation of these structural elements may cause changes in electrostatic charge and surface tension which result in increased permeability. The hemolytic and antitumor cell properties of protamine could not be segregated from its animal toxicity. Despite formalinization to a degree which eliminated the former, the compound remained quite toxic to mice and rabbits.