Detection of somatic coliphages through a bioluminescence assay measuring phage mediated release of adenylate kinase and adenosine 5'-triphosphate.

The feasibility of detecting somatic coliphages by phage infection of Escherichia coli WG5 and measurement of phage propagation by the lysis mediated release of the bacterial host adenylate kinase (AK) and adenosine 5'-triphosphate (ATP) detected by a bioluminescent signal was evaluated. After 2h of incubation, all cultures infected with reference bacteriophage phiX174 showed a significant increase in the bioluminescent signal, even with number of phages as low as less of 10 plaque forming units (PFU). Naturally occurring somatic coliphages ensured a significant bioluminescent signal after 3h of infection when >10 PFU were inoculated. These results indicate that an easy and reliable method to detect low numbers of coliphages in less than 3h is feasible.

Microbial indicators and pathogens: removal, relationships and predictive capabilities in water reclamation facilities.

Four water reclamation facilities in north-eastern Spain were monitored over 2 years to determine the occurrence and concentrations of a set of microbial indicators (total coliforms, Escherichia coli, enterococci, spores of sulphite reducing clostridia, somatic coliphages, F-specific RNA phages, phages infecting Bacteroides fragilis strain RYC2056 and phages infecting Bacteroides tethaiotaomicron strain GA-17), and two selected pathogens (cytopathogenic enteroviruses and viable Cryptosporidium oocysts). The indicator (survival) and index (presence) functions of the various indicators tested were evaluated through the wastewater treatments. The inactivation pattern of all groups of bacteriophages tested was closer to the inactivation of enteroviruses than to the inactivation of the conventional bacterial indicators tested. The inactivation of sulfite reducing clostridia spores and bacteriophages more closely approximates the reduction of viable Cryptosporidium than do the conventional bacterial indicators. We observed neither index functions nor a predictive relationship between any of microbial indicators and viable Cryptosporidium oocysts. In contrast, several regression models (r>0.6) and discriminant functions (67-88% well classified samples) based mostly on numbers of bacteriophages were able to predict both the presence and concentrations of enteroviruses. A combination of both bacterial and bacteriophage indicators seem to be the best choice for ensuring the microbial quality of reclaimed water.