液態培養結合ATP檢測用於評估自來水生物穩定性

Abstract

供水系統中的生物穩定性是重要的生物安全指標，培養基培養法檢測自來水中的總菌數無法精確反應加氯的自來水系統生物穩定性。隨著技術進步，三磷酸腺苷(adenosine triphosphate , ATP)檢測逐漸成熟，相較於傳統塗盤法更靈敏方便且能在短時間內獲得結果。本研究旨在開發一種液態培養方法，並利用三磷酸腺苷(ATP)進行微生物活性檢測取代塗盤法，以提高傳統自來水總菌數檢測方法的檢出率。首先，針對台灣自來水，使用國內外常用的總異營菌數培養基塗盤法進行比較，以瞭解不同培養基檢出量的差異。結果顯示R2A培養基所獲得的菌落數較高；後續研究則採用R2A培養液加入自來水樣品中進行液態培養之實驗。利用液態培養方法結合ATP檢測，評估液態培養替代塗盤法之可行性。利用液態培養進行最大生長速率及最大生長量之實驗檢測，所獲之ATP濃度與初始菌落數進行線性回歸，結果分別是(R^2=0.37)、(R^2=0.71)，顯示利用最大生長量的培養方法可以有效的提高ATP濃度與R2A菌落數間的關係。但利用T-TEST、Wilcoxon檢定結果顯示最大生長量所獲得之公式並無法有效的利用ATP濃度來推估初始菌落數，但在加氯後的自來水中經過液態培養方法後，可以有效的提高ATP濃度與菌落數間的關係。因此，若要以ATP做為消毒後的自來水之生物活性檢測，仍需要有更多的研究來進行補強。

Biostability in water supply systems is an important microbiological safety indicator. Traditional culture-based methods that measure total bacterial counts in tap water cannot accurately reflect the biostability of chlorinated water systems. With technological advancements, adenosine triphosphate (ATP) testing offering a more sensitive and convenient alternative to the traditional plate count method and provides results in a shorter time. This study aims to develop a different type of liquid culture method and use ATP to measure microbial activity, replacing the plate count method to improve the detection rate of traditional total microbial count methods in tap water. Initially, the study compared the plate count methods for heterotrophic bacteria with Taiwan's tap water to determine the differences between R2A and PCA media. The results indicated that the R2A medium yielded higher colony counts; subsequent experiments involved adding tap water samples to R2A broth for liquid culturing. This liquid culturing method, combined with ATP testing, was evaluated for its feasibility to replace the plate culture method. Experiments measuring maximum growth rates and biomass using liquid culture showed that ATP concentrations correlated with initial colony counts with linear regression results of R^2=0.37 and R^2=0.71, respectively. The results demonstrated that using maximum biomass culture methods could effectively enhance the relationship between ATP concentration and R2A colony counts. However, T-TEST and Wilcoxon test indicated that the formulas obtained from maximum biomass could not effectively use ATP concentration to estimate initial colony counts. However, in chlorinated tap water, the liquid culture method still effectively improved the relationship between ATP concentration and colony counts. Therefore, further research is needed to strengthen the use of ATP as a measure of biological activity in disinfected tap water.