The use of hollow fiber dialysis filters operated in axial flow mode for recovery of microorganisms in large volume water samples with high loadings of particulate matter

Abstract

Ultrafiltration concentration of microorganisms in large volume water samples containing high levels of particulate matter was evaluated in a proof of concept study. The organisms tested were Bacillus atrophaeus subspecies globigii spores and MS2 bacteriophage. To produce the large volume samples, fresh water sediment of a known particle size was added to 51 l of tap water. Five different concentrations of particulate matter were studied: 0, 50, 100, 150 and 750 mg solids/l. The concentration procedure used a dialysis filter as the ultrafilter configured for axial flow, either with or without recirculation. The target number of organisms spiked was 1 × 10⁵ of either spores or bacteriophage per 51 l. After concentration, the filters were dissected to retrieve the fibers which were then washed using surfactant solution which was then analyzed for the target organisms. Two washes of the filter fibers were carried out sequentially. For axial flow with recirculation, the first wash produced statistically greater recovery of B. globigii spores (26-40% of spike) compared to the second wash (8-13% of spike). Total recovery (the sum of the recoveries for the first and second washes) ranged from 35 to 53%. Recovery increased as the solids level increased from 0 to 150 mg solids/l. Recovery at the 100 and 150 mg solids/L loadings was statistically higher at the P < .05 level than recovery at 0 mg/L solids. At 150 mg solids/L, axial flow without recirculation (dead end) yielded lower recovery than axial flow with recirculation, however the difference was not significant at the P < .05 level. Recovery of B. globigii at 750 mg solids/L averaged 38% using dead end axial flow. The average recovery of MS2 bacteriophage was 45% at a solids concentration of 150 mg/L using axial flow with recirculation. PhiX174 and Phi8 were also studied, however these bacteriophage appeared to be inactivated in the matrix of concentrated wash water. One hundred liters of water containing 750 mg solids/L was concentrated using dead end axial flow, and only minimal problems with filter clogging were observed. Results described herein suggest axial flow ultrafiltration is an effective concentration method for microorganisms in water containing high levels of particulate matter.

Figure 1.

Tangential Flow HFUF Process

Figure 2.

HFUF Modified for Axial Flow

Figure 3.

Screen Shot of Software Used to Control the Automated Water Sample Concentrator

Figure 4.

Dead End Ultrafiltration Configured for Axial Flow

Figure 5.

Hollow Fiber Dialysis Filter Used in the Axial Flow Configuration. Ready for Dissection

Figure 6.

Percent Recovery of B. globigii spores. Axial Flow with Recirculation. Target Spike = 100,000 spores/51 liters.