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Pathogen Removal in Constructed Wetlands
Published in Donald A. Hammer, Constructed Wetlands for Wastewater Treatment, 2020
Richard M. Gersberg, R. A. Gearheart, Mike Ives
Water is collected in clean 1–liter Nalgene bottles and concentrated by membrane filtration through 0.45-m Gelman GN-6 filters. The filter is placed in a flask containing 50–100 mL of LSR broth. The flask is vigorously aerated (bubbled air was used, but a shaking water bath would work as well) at 35°C for 1 hour to starve the Salmonella and allow the other bacteria present to begin growth. Ampicillin is then added to a final concentration of 1000 mg/mL from a sterile solution in Davis minimal broth prepared the same day. Incubation is continued with aeration for 15 minutes. The cells are harvested by membrane filtration of the broth. The filter is then rinsed thoroughly in sterile buffer or minimal broth to remove the ampicillin, placed face up on XLD agar, and incubated for 24–36 hours at 41.5°C.
Internalisation of Salmonella spp. by Typha latifolia and Cyperus papyrus in vitro and implications for pathogen removal in Constructed Wetlands
Published in Environmental Technology, 2022
Richwell Alufasi, Wilson Parawira, Alexandros I. Stefanakis, Phiyani Lebea, Ereck Chakauya, Walter Chingwaru
The concentration of Salmonella spp. in the hydroponic medium was determined daily for a period of 12 days. Sample wastewater was taken from each hydroponic container every morning at 8am and serially diluted, by the factor of 10, to determine the concentration of Salmonella spp. according to a modified protocol of Bernstein et al. [31]. Briefly aliquots (100 µL) of the serially diluted samples were spread on XLD agar (Himedia Laboratories, India), in four replicates. The plates were incubated at 37°C for 48–72 h. Red, transparent colonies with black centres on XLD agar indicated the presence of Salmonella spp.