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M D ORLANDO

Publications and source records attributed to M D ORLANDO.

2 recordsLinked to original sources

SINGLE-DOSE ASSAY TECHNIQUE FOR VARIOLA VIRUS.

A biological assay for variola virus was needed that would (i) require a minimum of time and (ii) have acceptable precision. Preliminary titrations made in 11-day-old embryonated eggs and in suckling mice (6 to 24 hr of age) demonstrated a linear relationship between the concentration of variola virus injected and the mean reciprocal time to death (MTD) of both hosts. This linear response indicated that the injection of a single dose of virus suspension and the measurement of the MTD should result in an acceptable assay method. Seven replicate samples of liquid preparations (20% chorioallantoic membrane in Heart Infusion Broth) and the freeze-dried material obtained with these suspensions were assayed in triplicate for pock infectious units and for MTD. The variance of the pock counts was far greater than was expected from a Poisson distribution, and coefficients of variation ranged from 25 to 61%. Variances obtained with the single-dilution assay were all far below that expected from a Poisson distribution, and coefficients of variations ranged from 5.2 to 13.6%. The use of the MTD assay resulted in a saving of time, a saving in the number of hosts necessary per assay, and increased precision.

Animals↗

Viability and estimation of shelf-life of bacterial populations.

Mathematical concepts associated with the exponential and probit models are developed, and the similarity of the two methods is discussed. Because of its greater flexibility in design, the probit method was used to estimate the shelf-life for four bacterial populations, wet and dry spores of Bacillus anthracis and wet and dry cells of Pasteurella tularensis. On the basis of data gained by storing these organisms at high temperature, the probit method was used to predict the time at which 50% viability would occur for cells stored at 3 C. A plane passing through a three-space showing change in percentage viability of bacteria was formulated by a multiple regression method. With this functional technique, the percentage viability, expressed as a probit, was linearily related to a logarithm of storage time and storage temperature. The use of this method to study the effect of controlled variables on the viability of cells is demonstrated by comparing the effect of viability associated with three additives used prior to drying. The results of the test gave shelf-life estimates which were too low for all additives; however, the order of stability was ranked properly as confirmed by long-term tests.

Bacillus anthracis↗