Abstract: The present study was under taken to determine the relationship between spoilage factors of S. putrefaciens at different storage temperature and time in fresh whiting-fish with experimental treatment. The whiting fish samples taken from the Kefken region of Kand ra district in Kocaeli. Two groups were separated as control and test group of whiting-fish. Control group was divided into two parts. First part was stored at 0°C in 9 days and within this period, it was analysed in 1st, 3rd, 5th, 7th and 9th days of storage. Second part was also stored at -18°C for 6 months after freezed at -35°C. Test group was also divided into two groups and stored at the same conditions after contaminated with S. putrefaciens (Ref. No: CIP 80.40, Inst. Pasteur) at the level of 102 cfu g-1 and then all group samples analysed for total viable count, Enterobactericeae, Psycrophilic bacteria, Pseudomonas spp. S. putrefaciens, TMA amount and pH level. As S. putrefaciens average count was determined at under the detectable level (log10 < 2.0 cfu g-1) and 2.74 mg/100 g of TMA amountincontrol group, it determined at log10 3.66 cfu g-1and 5.59 mg/100 gof TMA amount in test group at 0°C storage temperature in 3rd day. It was determined that the differences became greater in the last day of storage (9th day). As S. putrefaciens average level at log10 8.6 cfu g-1 and 33.11 mg/100 g TMA amount in test group, it was found under the detectable level (log10 < 2.0 cfu g-1) and 12.04 mg/100 g TMA amount in control group. Therefore, S. putrefaciens had great effect on to spoilage of fish and TMA formation