益生菌在調節腦腸軸和影響神經功能的能力正日益受到重視。癲癇通常採用抗癲癇藥物（ASMs）治療，但許多患者對ASMs具有抗藥性或出現不良反應，這使得尋找更安全治療方案的必要性提升。本研究利用斑馬魚幼魚來篩選的抗癲癇作用益生菌。我們建立了一套標準化的實驗模型，明確了菌株劑量、餵食方案和幼魚發育階段。使用戊四唑（PTZ）誘導幼魚出現癲癇狀態，並即時分析幼魚的癲癇游泳行為。在測試的22株益生菌及其熱處理後的副生元中，有13株菌株顯著降低了PTZ誘導的癲癇發作。我們選擇BBM002及其熱處理形式（BBM002 HT）進行機制研究。在我們的研究發現，BBM002的完整的細胞壁（ICW）是介導抗癲癇作用的關鍵結構成分。在營養調控實驗中，高脂飲食會減弱益生菌的功效，而誘導性葉酸缺乏則會消除其對癲癇的抑製作用。相反，補充維生素B6的活性形式－磷酸吡哆醛（PLP）可增強BBM002和BBM002 HT的抗癲癇作用。這些發現支持BBM002和BBM002 HT有望成為控制癲癇的輔助治療方案，並證實飲食和代謝狀態對益生菌介導的抗癲癇作用具有調控能力。本研究可進一步去深入探討益生菌與營養物質在腦腸軸上的相互作用機制，並可支持發展腸道菌叢為主的癲癇治療策略。

Probiotics are increasingly recognized for their ability to modulate the gut–brain axis and influence neurological function. Epilepsy is commonly treated with anti-seizure medications (ASMs); however, many patients remain drug-resistant or experience adverse effects, underscoring the need for safer therapeutic alternatives. In this study, zebrafish larvae were used to screen probiotics for anti-seizure effect. A standardized protocol was established defining strain dosage, feeding schedule, and larval developmental stages. Seizure-like activity was induced by pentylenetetrazol (PTZ), and larval swimming behavior was analyzed in real time. Among the 22 probiotic and heat-treated parabiotic strains tested, 13 significantly reduced PTZ-induced seizures. BBM002 and its heat-treated form (BBM002 HT) were selected for mechanistic studies. The intact cell wall was identified as the critical structural component mediating anti-seizure effects. Nutritional modulation revealed that high-fat diet feeding attenuated probiotic efficacy, while inducible folate deficiency abolished seizure suppression. In contrast, supplementation with pyridoxal 5’-phosphate (PLP), the active form of vitamin B6, enhanced the anti-seizure effect of BBM002 HT. These findings identify BBM002 and BBM002 HT as promising adjunctive treatment for seizure control and demonstrate that dietary and metabolic states regulate probiotic-mediated anti-seizure effects. This study provides mechanistic insights into probiotic–nutrient interactions along the gut–brain axis and supports the development of microbiota-based therapeutic strategies for seizure.

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