COMPARATIVE STUDY OF SHELL SWAB AND SHELL CRUSH METHODS FOR THE RECOVERY OF SALMONELLA FROM SHELL EGGS

摘要

Swabbing is the standard methodology for the recovery of resident microorganism from shell eggs in Japan. A comparative study of shell swab (SW) and shell crush (CR) techniques was performed to recover the laboratory-inoculated Salmonella from shell eggs. It was found that the recovery of Salmonella by CR methods was significantly higher (4.5-7.5 log cfu-egg) than that of SW methods (3.1-6.3 log cfu-egg). However, analyses with quantitative real-time polymerase chain reaction (invA as a target gene), fluorescent microscopic and quantitative analyses with a Live-Dead BacLight bacterial viability kit revealed that not all of the inoculated Salmonella spp. populations were recovered as intact cells by either method. The chemiluminescent bacterial viability assay showed that chemiluminescence intensity (CI) began to increase after 30min in CR samples; on the other hand, SW samples did not show any increase in CI for 2h. These results suggest that SW might cause more damage and lethality to cells than CR.In addition, to determine the most appropriate method for recovering resident aerobic bacteria, coliforms and Salmonella spp from shell eggs, 4,000 commercial eggs were collected and sampled by shell rinse (SR) and CR techniques using phosphate-buffered saline (PBS) warmed to different temperatures. PBS at 37C was found to be the best recovery solution and temperature, respectively, for recovering aerobic microorganisms from shell eggs by both methods and the CR methods recovered a higher population than did the SR methods (4.9 versus 5.8 log cfu-egg for SR and CR methods, respectively; n=500 eggs-method).Therefore, the CR method along with recovery buffer (PBS) at 37C could be an effective technique for the recovery of microorganisms from post-processed shell eggs. There is a need to develop a rapid and highly sensitive method for the recovery of microorganisms from shell eggs. Such recovery methods are also useful for evaluating the efficacy of newly developed shell egg disinfection techniques. Many methods involving rinsing, swabbing, and crushing of shell eggs have been reported; however, we performed a comparative study of the method used to recover the Salmonella from shell eggs. We found that the shell crush method (CR) was superior to the shell swab method (SW) for the recovery of Salmonella spp., and phosphate-buffered saline (PBS) at 37C was found to be the best recovery solution and temperature, respectively, for recovering microorganisms from shell eggs by both methods. Therefore, the CR method along with recovery buffer (PBS) at 37C could be an effective technique for the recovery of microorganisms from post-processed shells. Use of this method could be recommended for the microbial evaluation of post-processed shell eggs in industries.