Rapid enumeration of physiologically active bacteria in purified water used in the pharmaceutical manufacturing process.
Physiologically active bacteria in purified water used in the manufacturing process of pharmaceutical products were enumerated in situ. Bacteria with growth potential were enumerated using the micro-colony technique and direct viable counting (DVC), followed by 24 h of incubation in 100-fold diluted SCDB (Soybean Casein Digest Broth) at 30 degrees C. Respiring and esterase-active bacteria were detected by fluorescent staining with 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and 6-carboxyfluorescein diacetate (6CFDA), respectively. A large number of bacteria in purified water retained physiological activity, while most could not form colonies on conventional media. The techniques applied in this study enabled bacteria to be counted within 24 h so results could be available within one working day. These rapid and convenient techniques should be useful for the systematic monitoring of bacteria in water used for pharmaceutical manufacturing.
Kawai
M
M
Graduate School of Pharmaceutical Sciences, Osaka University, Japan.
Yamaguchi
N
N
Nasu
M
M
D001431:Bacteriological Techniques; D016956:Burkholderia cepacia / Q000201:enzymology / Q000254:growth & development*; D002839:Chromans; D003470:Culture Media; D004345:Drug Industry / Q000592:standards*; D004950:Esterases / Q000378:metabolism; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005618:Fresh Water / Q000382:microbiology; D010100:Oxygen / Q000378:metabolism; D013212:Staphylococcus epidermidis / Q000201:enzymology / Q000254:growth & development*; D018508:Water Purification; D014881:Water Supply / Q000592:standards*
D002839:Chromans; D003470:Culture Media; D005452:Fluoresceins; D005456:Fluorescent Dyes; C059114:2-(3-chloropropyl)-2,5,7,8-tetramethyl-6-chromanol; C093616:6-carboxyfluorescein diacetate; D004950:Esterases; D010100:Oxygen