In this study, we developed Rapid Enrichment Broth for Vibrio parahaemolyticus (REB-V), a broth capable enriching V. parahaemolyticus from 100 CFU/mL to 106 CFU/mL within 6 hours, which greatly facilitates the rapid detection of V. parahaemolyticus. Using a modified Gompertz model and response surface methodology, we optimized supplement sources to rapidly enrich V. parahaemolyticus. The addition of 0.003 g/10 mL of D-(+)- mannose, 0.002 g/10 mL of L-valine, and 0.002 g/10 mL of magnesium sulfate to 2% (w/v) NaCl BPW was the most effective combination of V. parahaemolyticus enrichment. Optimal V. parahaemolyticus culture conditions using REB-V were at pH 7.84 and 37oC. To confirm REB-V culture efficiency compared to 2% (w/v) NaCl BPW, we assessed the amount of enrichment achieved in 7 hours in each medium and extracted DNA samples from each culture every hour. Real-time PCR was performed using the extracted DNA to verify the applicability of this REB-V culture method to molecular diagnosis. V. parahaemolyticus was enriched to 5.452±0.151 Log CFU/mL in 2% (w/v) NaCl BPW in 7 hours, while in REB-V, it reached 7.831±0.323 Log CFU/mL. This confirmed that REB-V enriched V. parahaemolyticus to more than 106 CFU/mL within 6 hours. The enrichment rate of REB-V was faster than that of 2% (w/v) NaCl BPW, and the amount of enrichment within the same time was greater than that of 2% (w/v) NaCl BPW, indicating that REB-V exhibits excellent enrichment efficiency.

본 연구에서는 축산식품인 편육을 대상으로 황색포도상 구균의 성장예측모델을 개발하였다. 편육에서 황색포도상 구균의 성장패턴은 4, 10, 20, 37oC의 보관온도에서 측정 되었으며, 황색포도상구균은 각각의 저장 온도에서 모두 증가하는 것으로 나타났다. 편육에 오염된 황색포도상구 균의 생육결과를 토대로 Baranyi model을 이용하여 유도 기(LPD)와 최대성장률(μmax)을 산출한 결과, 유도기는 4, 10, 20, 37oC에서 212.81, 79.67, 3.12, 2.21 h으로 온도에 반비례한 것으로 나타났고 최대성장률은 같은 보관온도에 서 0.004, 0.009, 0.130, 0.568 log CFU/g/h으로 온도에 비 례한 것으로 조사되었다. 2차 모델은 μmax의 경우, square root model, LPD는 polynomial equation을 사용하여 산출 하였고, 개발한 모델의 적합성을 평가하기 위해 통계적 지 표인 RMSE 값을 계산한 결과, 비교적 0에 가까운 0.42로 도출되어 모델이 적합한 것으로 확인되었다. 따라서 개발된 모델이 편육에 대한 황색포도상구균의 성장 예측모델로 사용 가능하다고 판단되어지며, 편육에서의 식중독을 예방하고 미생물학적 위생관리기준을 설정하는데 기초자 료로 활용될 수 있을 것으로 사료된다.

본 연구에서는 멸균처리공정이 없는 돈육 포장공정을 대상으로 작업장에서 직접 분리한 야생균주인 Salmonellaspp. KSC101를 작업장의 온도와 시간을 주요 변수로 하여, 이들 현장에서의 Salmonella spp. KSC101의 성장 특성을 파악하고, 이를 수학적으로 예측할 수 있는 모델을 개발하였다. 돈육포장공장 현장을 반영하여 온도는 0, 5, 10,15, 20oC로, 시간은 0, 1, 2, 3시간으로 하였으며, 0oC와5oC에서는 성장이 발생하지 않았으며, 10oC, 15oC, 20oC에서는 약간의 성장이 있었으나 증가수준은 평균 0.34 logCFU/g정도였고, 20oC에서 성장율이 더 높았으나 15oC와는 통계적으로는 유의하지 않았다(p < 0.05). 하지만 PMP와 비교시 야생균주인 Salmonella spp. KSC101의 성장이더 빠른 것으로 나타났다. 이들 실험결과를 바탕으로 1차모델은 Gompertz 4 parameter식을, 2차 모델은 Exponentialdecay식을 이용하여 성장예측모델을 개발하였으며, R2값은0.99이상으로 나타났다. 개발된 모델에 대한 검증으로RMSE를 이용하였으며, 값이 0.103으로 양(+)의 방향으로약간 초과 예측하는 것으로 나타났으나, 최종적으로 실험값과 예측값이 ± 0.5 log cfu/g 내에서 일치하고 있어, 본연구에서 개발된 모델은 추후 냉장돈육 포장공정에서 위생관리기준 설정에 대한 과학적 근거자료로 활용할 수 있을 것이다.

Predictive mathematical models were developed for predicting the kinetics of growth of Listeria monocytogenes in smoked salmon, which is the popular ready-to-eat foods in the world, as a function of temperature (4, 10, 20 and 30℃). At these storage temperature, the primary growth curve fit well (r² = 0.989~0.996) to a Gompertz equation to obtain specific growth rate (SGR) and lag time (LT). The Polynomial model for natural logarithm transformation of the SGR and LT as a function of temperature was obtained by nonlinear regression (Prism, version 4.0,GraphPad Software). Results indicate L. monocytogenes growth was affected by temperature mainly, and SGR model equation is 365.3-31.94*Temperature+0.6661*Temperature^2 and LT model equation is 0.1162-0.01674*Temperature+0.0009303*Temperature^2. As storage temperature decreased 30℃ to 4℃, SGR decreased and LT increased respectively. Polynomial model was identified as appropriate secondary model for SGR and LT on the basis of most statistical indices such as bias factor (1.01 by SGR, 1.55 by LT) and accuracy factor (1.03 by SGR, 1.58 by LT).

In this study, predictive mathematical models were developed to predict the kinetics of Listeria monocytogenes growth in the mixed fresh-cut vegetables, which is the most popular ready-to-eat food in the world, as a function of temperature (4, 10, 20 and 30oC). At the specified storage temperatures, the primary growth curve fit well (r2 = 0.916~0.981) with a Gompertz and Baranyi equation to determine the specific growth rate (SGR). The Polynomial model for natural logarithm transformation of the SGR as a function of temperature was obtained by nonlinear regression (Prism, version 4.0, GraphPad Software). As the storage temperature decreased from 30oC to 4oC, the SGR decreased, respectively. Polynomial model was identified as appropriate secondary model for SGR on the basis of most statistical indices such as mean square error (MSE = 0.002718 by Gompertz, 0.055186 by Baranyi), bias factor (Bf = 1.050084 by Gompertz, 1.931472 by Baranyi) and accuracy factor (Af = 1.160767 by Gompertz, 2.137181 by Baranyi). Results indicate L. monocytogenes growth was affected by temperature mainly, and equation was developed by Gompertz model (−0.1606 + 0.0574*Temp + 0.0009*Temp*Temp) was more effective than equation was developed by Baranyi model (0.3502 − 0.0496*Temp + 0.0022*Temp* Temp) for specific growth rate prediction of L. monocytogenes in the mixed fresh-cut vegetables.