Methodology for enumeration of coliphages in foods.
The effects of eluent composition, pH, and chaotropic agents on the recovery of T2, MS2, and indigenous coliphages from various foods were investigated. Additionally, methods of sample suspension and clarification were evaluated for coliphage recovery and application to various foods. Clarified sample suspensions were assayed for coliphages with a modified agar layer technique and appropriate Escherichia coli hosts. Centrifugation and polypropylene mesh filtration were more rapid and effective than glass wool filtration for clarification of sample suspensions and subsequent recovery of coliphages. Blending, stomaching, and shaking procedures were generally comparable for sample liquefaction and release of coliphages from foods. Complex basal eluents, EC medium and 1% casein, were generally more effective than a less complex eluent, phosphate buffer, for elution of coliphages from foods. For most foods, incorporation of sodium chloride or chaotropic agents, i.e., sodium trichloroacetate, urea, Tween 80, Triton X-100, and sodium nitrate, into basal eluents did not enhance recovery of coliphages. Indigenous coliphage recovery was not affected by sample suspension pH over a range of 6.0 to 9.0. With an optimal procedure, i.e., EC medium eluent, blending, and centrifugation, the recovery of T2 and MS2 ranged from 48 to 81% and from 58 to 100%, respectively, depending on the food type.
D000818:Animals; D001431:Bacteriological Techniques; D002417:Cattle; D002498:Centrifugation; D002645:Chickens; D003090:Coliphages / Q000254:growth & development / Q000302:isolation & purification*; D003470:Culture Media; D004926:Escherichia coli / Q000032:analysis; D005374:Filtration; D005516:Food Microbiology; D005519:Food Preservation; D006863:Hydrogen-Ion Concentration; D008460:Meat; D013552:Swine; D014675:Vegetables
D003470:Culture Media