Using a Blender to Assess the Microbial Density of Encapsulated Organisms

There are specific NASA requirements for source-specific encapsulated microbial density for encapsulated organisms in non-metallic materials. Projects such as the Mars Science Laboratory (MSL) that use large volumes of non-metallic materials of planetary protection concern pose a challenge to their bioburden budget. An optimized and adapted destructive hardware technology employing a commercial blender was developed to assess the embedded bioburden of thermal paint for the MSL project. The main objective of this optimization was to blend the painted foil pieces in the smallest sizes possible without excessive heating. The small size increased the surface area of the paint and enabled the release of the maximum number of encapsulated microbes. During a trial run, a piece of foil was placed into a blender for 10 minutes. The outside of the blender was very hot to the touch. Thus, the grinding was reduced to five 2-minute periods with 2-minute cooling periods between cycles. However, almost 20% of the foil fraction was larger (>2 mm). Thus, the largest fractions were then put into the blender and reground, resulting in a 71% increase in particles less than 1 mm in size, and a 76% decrease in particles greater than 2 mm in size. Because a repeatable process had been developed, a painted sample was processed with over 80% of the particles being <2 mm. It was not perceived that the properties (i.e. weight and rubber-like nature) of the painted/foil pieces would allow for a finer size distribution. With these constraints, each section would be ground for a total of 10 minutes with five cycles of a 2-minute pulse followed by a 2-minute pause. It was observed on several occasions that a larger blade affected the recovery of seeded spores by approximately half an order of magnitude. In the standard approach, each piece of painted foil was aseptically removed from the bag and placed onto a sterile tray where they were sized, cut, and cleaned. Each section was then weighed and placed into a sterile Waring Laboratory Blender. Samples were processed on low speed. The ground-up samples were then transferred to a 500-mL bottle using a sterile 1-in. (.2.5-cm) trim brush. To each of the bottles sterile planetary protection rinse solution was added and a modified NASA Standard Assay (NASA HBK 6022) was performed. Both vegetative and spore plates were analyzed.