Development and Validation of a Cultural Method for the Detection and Isolation of Salmonella in Cloves

Detection of Salmonella in some spices, such as cloves, remains a challenge due to their inherent antimicrobial properties. The purpose of this study was to develop an effective detection method for Salmonella from spices using cloves as a model. Two clove varieties, Ceylon and Madagascar, were used in the study. Cloves were inoculated with Salmonella enterica subsp. enterica serotypes Montevideo, Typhimurium, or Weltevreden at about 1, 3, or 6 log CFU/25 g. Two test portion sizes, 10 and 25 g, were compared. After adding Trypticase soy broth (TSB) to the weighed cloves for preenrichment, three preenrichment methods were compared: cloves were left in the TSB for 24 h during preenrichment (PreE1), or the cloves-TSB mixture was shaken vigorously for 30 s (PreE2) or 60 s (PreE3), and the decanted material was transferred to a new bag for 24 h of preenrichment. The rest of the procedures were carried out according to the U.S. Food and Drug Administration Bacteriological Analytical Manual (BAM). At the low inoculation level (<1 log CFU/25 g), the detection rate was low across the three preenrichment methods, with the highest for PreE3 and lowest for PreE1. At the medium and high inoculation levels (3 and 6 log CFU/25 g), all samples from PreE2 and PreE3 were positive for Salmonella , whereas PreE1 produced only 12 positive samples from the 48 samples at the medium inoculation level and 38 positive samples from the 48 samples at the high inoculation level. Therefore, PreE3 with 25 g of cloves per sample was more effective than the other two tested methods. This newly designed method was then validated by comparing with the BAM method in six trials, with each trial consisting of 40 test samples. The results showed that PreE3 detected Salmonella from 88 of 120 inoculated test samples compared with only 31 positive from 120 test samples with the BAM method. Thus, our newly designed method PreE3 was more sensitive and easier to operate than the current BAM method for detection of Salmonella in cloves.