The redox dye 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) was employed for direct epifluorescent microscopic enumeration of respiring bacteria in environmental samples. Oxidized CTC is nearly colorless and is nonfluorescent; however, the compound is readily reduced via electron transport activity to fluorescent, insoluble CTC-formazan, which accumulates intracellularly. Bacteria containing CTC-formazan were visualized by epifluorescence microscopy in wet-mount preparations, on polycarbonate membrane filter surfaces, or in biofilms associated with optically opaque surfaces. Counterstaining of CTC-treated samples with the DNA-specific fluorochrome 4',6-diamidino-2-phenylindole allowed enumeration of active and total bacterial subpopulations within the same preparation. Municipal wastewater, groundwater, and seawater samples supplied with exogenous nutrients yielded CTC counts that were generally lower than total 4',6-diamidino-2phenylindole counts but typically equal to or greater than standard heterotrophic (aerobic) plate counts. In unsupplemented water samples, CTC counts were typically lower than those obtained with the heterotrophic plate count method. Reduction of CTC by planktonic or biofilm-associated bacteria was suppressed by formaldehyde, presumably because of inhibition of electron transport activity and other metabolic processes. Because of their bright red fluorescence (emission maximum, 602 nm), actively respiring bacteria were readily distinguishable from abiotic particles and other background substances, which typically fluoresced at shorter wavelengths. The use of CTC greatly facilitated microscopic detection and enumeration of metabolically active (i.e., respiring) bacteria in environmental samples.

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Clogging due to the accumulation of suspended solids is a major constraint that limits the capacity of Orange County Water District's (OCWD) surface water recharge system. In order to decrease clogging and increase system capacity, OCWD is testing the ability of riverbed filtration to reduce suspended solids concentrations and improve recharge rates. Riverbed filtration is achieved through a shallow subsurface collector system placed approximately one meter below the surface. Filtered water from the collector system is conveyed by gravity to the receiving recharge basin. Initial results show that riverbed filtration is highly effective in removing suspended solids in the recharge water, which in turn also greatly increases the recharge capacity of the receiving basin. Some other water quality benefits are also achieved. Data collected thus far indicate that it will be cost-effective to use this approach at a larger scale to capture and recharge increased quantities of storm flow obtained from the Santa Ana River.

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