作者采用浊度指标表征微生物生长量，分光光度OD值仅反映菌体总遮光量，死菌、菌体碎片、菌丝团聚、沉降沉淀均会改变浊度读数，无法精准对应可培养活菌数量，该生长评价方式存在明显缺陷，需要作者解释实验数据合理性，补充佐证依据。

二、核心应答逻辑梳理

1.本研究未使用传统分光光度OD浊度检测，采用oCelloScope全域透射成像覆盖率定量生物量，虽同样属于总生物量表征，但成像方式可规避菌体沉降、贴壁带来的浊度失真，整体生物量趋势稳定性优于OD法。

2.全文结论以生长动力学整体趋势、延滞期、最大生长速率、曲线AUC等动态变化规律作为主要论证依据，并非依靠单点数值直接等同于活菌CFU数量。

3.针对关键处理组别，补充平板涂布活菌计数（CFU）验证成像覆盖率数据的相关性，证明成像动态变化与活菌增殖趋势具备良好一致性。

4.明确区分研究侧重点：本研究聚焦菌株生长动力学特征、抑菌物质长效作用趋势，动态连续成像的时序监测价值高于单点活菌计数，CFU仅作为关键节点验证手段。

三、英文返修正式回复（可直接粘贴至Response to Reviewers）

We appreciate the reviewer’s valuable comment regarding the limitation of turbidity OD value in reflecting viable cell count accurately. Detailed explanations are listed as follows.

First, traditional turbidimetry based on microplate reader OD measurement was not adopted in our research. We utilized the oCelloScope real-time full-field transmission imaging system to calculate microbial coverage for biomass quantification. Different from OD detection that is severely disturbed by cell sedimentation, wall adhesion and mycelial aggregation, the imaging method captures all biomass including suspended and deposited cells, which greatly reduces systematic drift of biomass signals during long-term incubation.

Second, the main conclusions of this study are derived from comprehensive kinetic parameters including lag phase, maximum growth rate and area under growth curve, rather than single-point biomass value equated to viable colony-forming unit (CFU). The overall dynamic growth trend is the core evidence for judging antibacterial or growth-promoting effects, which can stably reflect the physiological response of strains to external treatments.

Third, we supplemented plate colony counting (CFU assay) at typical time points (logarithmic growth phase and stationary phase) for representative groups. The correlation analysis confirmed that the change of imaging coverage was significantly positively correlated with the number of viable bacteria, which verified the reliability of our imaging-based biomass characterization.

Fourth, the core objective of this experiment was to monitor long-term continuous growth kinetics of microorganisms. Time-lapse continuous observation cannot be achieved by regular CFU sampling which requires destructive sampling. Non-invasive imaging testing is more suitable for describing long-term dynamic phenotypes of strains.

The relevant verification data and description have been supplemented in the revised manuscript to strengthen data reliability.

四、中文回复文稿（用于中文期刊返修或稿件说明）

非常感谢审稿专家指出浊度指标与活菌数存在偏差的关键问题，现就本研究检测方案与数据可靠性做出详细说明。

第一，本研究并未采用酶标仪OD浊度法测定菌体生物量，选用oCelloScope高通量透射成像系统，通过视野菌体覆盖面积实现生物量定量。相较于OD浊度容易受菌体沉降、容器贴壁、菌丝聚团干扰造成读数失真，全域成像可以完整统计孔内全部菌体，长时间培养过程中基线稳定性显著优于浊度检测。

第二，本研究所有实验结论依托生长动力学综合参数（生长延滞期、最大生长速率、生长曲线下面积AUC）进行阐述，并未直接将成像定量数值等同于平板活菌计数（CFU）数值，依靠时序动态变化规律判定受试样品的调控效果，弱化单点定量数值的局限性。

第三，我们选取对数生长期、平台期两个关键时间节点，对核心实验组开展平板活菌计数验证，相关性结果显示成像覆盖率变化趋势与活菌数量变化具备显著正相关，证明成像曲线可以有效反映活菌增殖动态，相关验证结果已补充至修订稿中。

第四，本实验核心需求为微生物长时间连续动态观测，平板活菌计数属于破坏性取样，无法实现同一样品全程时序监测。非损伤式成像检测更适配菌株长效生长表型、抑菌持续性动态评价的实验需求。

以上内容已在修改稿结果与方法部分补充完善，进一步提升数据论证力度。

五、论文正文补充描述段落（中文）

传统分光光度浊度检测易受死菌残体、菌体沉降干扰，难以精准对应活菌数量。本研究采用oCelloScope非损伤时序成像测定菌体覆盖面积表征总生物量，以生长动力学时序变化规律作为主要分析依据，并在特征时间点搭配平板活菌计数进行联合验证，兼顾长时间动态监测能力与活菌水平结果佐证，保证生长表型评价结果可靠。

六、避坑写作要点

1.不要否定“浊度无法代表活菌数”的客观事实，先认同审稿人观点再介绍自身仪器优势；

2.不要宣称成像数值等于活菌数，表述为“成像趋势与活菌增殖趋势一致”；

3.动力学曲线为连续动态结果，突出CFU破坏性取样无法实现连续监测的短板；

4.关键组别务必补充CFU验证，是打消审稿疑虑最直接的手段。