Sampling Size Optimization for Bioburden Density Estimation in Planetary Protection

Abstract

Planetary protection (PP) is a discipline that focuses on minimizing the biological contamination of spacecraft to ensure compliance with international policy. Precise estimation of bioburden - the total number of microbes in or on spacecraft hardware – and the bioburden density are of utmost importance for PP. Such estimation is the way concordance with requirements is demonstrated, and it is critical for quantifying the potential risk of inadvertently contaminating other planetary bodies. Although a suite of molecular techniques have been used to thoroughly characterize and profile the microbiome of various cleanroom environments and spacecraft, the gold standard remains the physical enumeration of microbes via culturing of samples directly taken from spacecraft and associated surfaces. However, due to technical, budgetary, and programmatic constraints, only a manageable portion (around 10%) of the entire spacecraft surface is directly sampled with cotton swabs or wipes. To generate the bioburden current best estimate (CBE) for components not directly verifiable, the accepted approach is to apply a NASA-defined bioburden estimate based on the components’ manufacturing or assembly environment. This approach utilizes a prespecified bioburden density estimation that applies a maximum value across the total surface area of the specified component. For hardware components that underwent similar assembly processes, an implied bioburden is adopted for all components, based on a direct verification of a representative component within the same lot. Once all components have a CBE, the bioburden estimates are generated. In previous publication [ 1], we have shown that statistical risks quantifying the accuracy of the estimates for sampled, prespecified, and implied components can be derived and ranked. For mean squared error (MSE) function, the risks are available analytically and hence a cost function can be obtained to optimize the risks with respect to the sampling area and sampling cost. Since the sampling area and sampling cost are two complimentary variables, their sum will have a well-defined minimum. This paper presents the multivariate optimization of the integrated risk of an empirical Bayes estimator to determine the optimal sampling schedule for a given number of components. It is assumed that given a number of components, N, the bioburden density for each component can either be sampled, implied, or prespecified. The multivariate optimization searches through different options to sample, imply or prespecify the bioburden density for a component, and account for the component’s surface area and cost of sampling. The idea of the optimization is based on the observation that the statistical risk of using an estimator is a monotonically decreasing function of the sampled area. The larger the sampled area, the lower the risk of using the estimator as the estimator becomes more and more accurate as the sampling area increases. On the other hand, the cost of sampling is monotonically increasing as the sampled surface grows. This makes the risk and total cost of sampling complimentary variables which can be counterbalanced to achieve an optimal overall value with respect to the sampled surface. In this paper, the integrated risk has been used to quantify the accuracy of the estimator. This risk has been selected because it depends on neither the true value of the parameter nor on the collected data. The cost of each sample was also available to obtain the total cost of sampling of N components. The paper will present the results based on computer-simulated data as well as the data collected during the InSight mission. The computer-simulated data have N components with randomly generated total areas and each component assigned to one of the three categories according to the method of estimating of bioburden density: sampled, implied, or prespecified. The cost of sampling is also available. The cost of sampling is estimated based on a cost model provided by the planetary protection group at JPL. For this paper, the overall cost was assumed to be a linear function of exposure. The optimization process finds the allocation of the components to the three categories that minimizes the tradeoff between integrated risk and total cost. For the InSight data, a set of components is selected representing all three categories, and optimization is performed to determine if the performed allocation was optimal or if a better allocation could have been obtained. To the best of our knowledge, this work is the first attempt not only perform an accurate estimation of bioburden density but also do it in an optimal way.