Concentration of Bacteriophages from Natural Waters

Abstract

The methods used for concentrating animal viruses from drinking water were found to be unsuitable for the concentration of bacteriophages from natural waters. The factors affecting recovery were investigated and a concentration procedure devised which is amenable to larger scale and field use. This procedure involves: (1) passage of the water through a sand filter; (2) removal of dissolved organic material with an anion exchange resin; (3) addition of MgCl2 to a final concentration of 5 times 10‐4m; (4) adjustment of the pH value to 3°8; (5) adsorption of the bacteriophages on to fibre glass and cellulose nitrate filters; (6) elution of bound phage with 3% (w/v) beef extract, and (7) concentration by ultrafiltration of the resulting eluates. Using this procedure a wide range of test bacteriophages was concentrated from 41 to 5 ml with recoveries ranging from 18–80%—concentration factors of 200–900 fold.

Virioplankton: Viruses in Aquatic Ecosystems

“…In particular, the inactivating effects of bacterioplankton and heat-labile dissolved compounds can be offset by the protective effect of viral adsorption to sediment and particulates suspended in the water column. Indeed, the general sorptive properties of viruses to suspended particulates in water (see the review by Bitton [26]) have been exploited in several methods for concentrating viruses from natural water samples (7,81,82,252,287,288,336). In controlled experiments, the addition of clay or sediment to water, prior to the addition of virus, lowered the decay rate of both enteroviruses (158,297) and coliphages (27,105).…”

A Portable Device for Concentrating Bacteriophages from Large Volumes of Freshwater

“…The large scale concentrator described here recovered test phages as efficiently as the small-scale system described previously (Seeley & Primrose 1979) and we consider that scale-up has been worthwhile. Indeed, the relative recoveries of the eight test phages (Table 1) were very similar to those achieved with the small-scale system.…”

“…The media, bacteria and bacteriophages used have been described previously (Seeley & Primrose 1979).…”

“…The pH was adjusted to 5.0 with 1 N HCI and then, using the p H meter in the expanded scale mode, reduced to 3.8 with 0-I N HCI. The acidified water was pumped through an acid-washed (Seeley & Primrose 1979) filter sandwich contained in the adsorbent filter holder. The sandwich comprised a 293 mm Metrigard GN6 membrane (Gelman Hawksley Ltd, Lancing, Sussex) protected by a Gelman 265 mm polypropylene spacer and a 293 mm Millipore AP20 prefilter.…”

“…Gerba et al (1978) found that the use of pleated membrane filter cartridges permitted up to 5Ooh of added poliovirus to be recovered from 190 I of secondary sewage effluent and 378 1 of estua;y water. In a previous communication (Seeley & Primrose 1979) we reported the appiication of some of the techniques of animal virus concentration to the recovery of a wide range of bacteriophage types from turbid pond water. We now report the application e?…”