1 Ganeden Inc., 5800 Landerbrook Drive, Suite 300, Mayfield Heights, OH 44124, USA.
2 Maastricht University - campus Venlo, Centre for Healthy Eating & Food Innovations, St. Jansweg 20, 5928 RC Venlo, the Netherlands.
Benef Microbes. 2019 Feb 8;10(1):77-87. doi: 10.3920/BM2018.0037. Epub 2019 Jan 29.
The aim of this study was to assess the germination, survival and metabolic activity of the probiotic Bacillus coagulans GBI-30, 6086 [GanedenBC] (BC30) in a dynamic, computer controlled in vitro model of the gastrointestinal (GI) tract, simulating human adults. Experiments were performed in the presence of a meal to maximise germination, due to the presence of germination-triggers. Both an upper GI tract (stomach and small intestine; TIM-1) and a colon model (TIM-2) were used, where material exiting TIM-1 was added to TIM-2. Spores of BC30 were introduced in the gastric compartment of TIM-1 and samples were taken immediately after the pylorus. Moreover, for 6 h, every hour the ileal efflux was collected and a subsample was plated for viable counts (spores and germinated cells). The remainder of the sample was fed to TIM-2, and after 24 h another sample was taken and tested for viable counts. In addition, samples were taken from the dialysates of the model and analysed using LC-MS/MS to determine bacterial metabolites and digestion products. Survival after transit through the gastric compartment was high (97%) and most cells were still in the spore form (76%). Survival after transit through TIM-1 was on average 51%, meaning that on average half of the orally provided spores was found back as cfu on the agar plates. Of these on average 93% were germinated cells and only 7% were spores. 24 h after the start of the experiments germination had increased in TIM-2 to 97% vegetative cells, and only 3% spores. No further loss of viability was observed in TIM-2. In terms of metabolic activity, increased levels of amino acids, dipeptides and citric acid cycle metabolites were found compared to experiments in the absence of BC30. In conclusion, BC30 spores germinate to a large extent (>90%) in the presence of germination triggers in the small intestine in a model that closely mimics the physiological conditions of human adults. Of the oral dose, as much as half of the cells survived transit through the upper GI tract, and based on the metabolite profile, these cells were metabolically active. Either these cells or the enzymes released from the dead cells aided in digestion of the meal. These insights help explain some of the observations in previous experiments, and support the understanding of the mechanism of action of the probiotic BC30.
本研究旨在评估益生菌凝结芽孢杆菌 GBI-30,6086[GanedenBC]（BC30）在动态、计算机控制的胃肠道（GI）体外模型中的发芽、存活和代谢活性，该模型模拟成人。由于存在发芽触发物，实验在进食时进行，以最大限度地促进发芽。使用了上胃肠道（胃和小肠；TIM-1）和结肠模型（TIM-2），其中从 TIM-1 中排出的物质被添加到 TIM-2 中。BC30 的孢子被引入 TIM-1 的胃腔中，在幽门后立即取样。此外，在 6 小时内，每小时收集回肠流出物的样本并进行活菌计数（孢子和发芽细胞）。其余的样本被喂给 TIM-2，24 小时后再次取样并进行活菌计数。此外，从模型的透析液中取样，并使用 LC-MS/MS 分析以确定细菌代谢物和消化产物。通过胃腔的转运后存活率很高（97%），大多数细胞仍处于孢子形式（76%）。通过 TIM-1 的转运后存活率平均为 51%，这意味着平均一半的口服提供的孢子在琼脂平板上作为 cfu 被发现。其中平均 93%为发芽细胞，只有 7%为孢子。实验开始 24 小时后，TIM-2 中的发芽率增加到 97%的营养细胞，只有 3%的孢子。在 TIM-2 中没有观察到进一步的生存能力损失。就代谢活性而言，与不存在 BC30 的实验相比，发现了氨基酸、二肽和柠檬酸循环代谢物水平的增加。总之，在小肠中存在发芽触发物的情况下，BC30 孢子在很大程度上（>90%）发芽，该模型非常接近成人的生理条件。在口服剂量中，多达一半的细胞通过上胃肠道转运存活下来，并且根据代谢物谱，这些细胞具有代谢活性。这些细胞或从死亡细胞释放的酶有助于消化膳食。这些见解有助于解释以前实验中的一些观察结果，并支持对益生菌 BC30 作用机制的理解。
