กิจกรรมการยับยั้งแบคทีเรียและการต้านไบโอฟิล์มของแอคติโนแบคทีเรีย ที่แยกจากดินรอบรากพืชสมุนไพรบางชนิด Antibacterial Activities and Antibiofilm Formation of Actinobacteria Isolated from Rhizospheric Soils of Some Medicinal Plants
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Abstract
Actinobacteria are an important group of Gram-positive bacteria capable to produce secondary metabolite with biological activities and has been used as antimicrobial agents and including antibiofilm formation. Actinobacteria are commonly found in soil, water, plant tissue and plant rhizosphere.The aim of this research was to investigate the antibacterial activity and antibiofilm formation of actinobacteria isolated from rhizospheric soils of three medicinal plants including Andrographis paniculata, Cannabis sativa and Mitragyna speciosa against Bacillus cereus TISTR 687, Staphylococcus aureus ATCC 27853, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa. 40 actinobacteria isolates were recovered from rhizospheric soils of A. paniculata 17 isolates, M. speciosa 12 isolates and C. sativa 11 isolates. Out of these, 11 isolates were found to inhibit at least one tested bacteria. There were three isolates namely F17, KC11 and KT01 exhibited high potential of antimicrobial activity and were selected for antibiofilm formation test using the microtiter plate (MTP) assay. The results showed that the isolate KT01 exhibited the highest antibiofilm activity. Cell free supernatant of isolate KT01 inhibited biofilm formation of B. cereus TISTR 687, S. aureus ATCC 27853 E. coli ATCC 25922 and P. aeruginosa, accounting for 11.69 ± 0.06%, 19.32 ± 0.05%, 23.57± 0.37% and 87.46 ± 0.07%, respectively. Based on 16s rDNA gene analysis, the isolate KT01 (1,499 bp) was most closely related to Streptomyces griseoincarnatus LMG 19316T (100% similarity). This result demonstrated that actinobacteria, isolated from the rhizospheric soils of medicinal plants, are source of bioactive compounds to inhibit pathogenic bacteria and antibiofilm.
บทคัดย่อ
แอคติโนแบคทีเรียเป็นแบคทีเรียแกรมบวกกลุ่มสำคัญที่สามารถผลิตสารเมทาบอไลต์ทุติยภูมิที่มีฤทธิ์ทางชีวภาพ และถูกนำมาใช้เป็นสารต้านจุลินทรีย์ แอคติโนแบคทีเรียสามารถพบได้ทั่วไปในดิน น้ำ เนื้อเยื่อพืชและในดิน รอบรากพืช งานวิจัยนี้มีวัตถุประสงค์เพื่อศึกษากิจกรรมการยับยั้งแบคทีเรียและการต้านไบโอฟิล์มของแอคติโนแบคทีเรียที่แยกจากดินรอบรากพืชสมุนไพร 3 ชนิด คือ ฟ้าทะลายโจร (Andrographis paniculata) กัญชา (Cannabis sativa) และกระท่อม (Mitragyna speciosa) ต่อการยับยั้งแบคทีเรียทดสอบ Bacillus cereus TISTR 687, Staphylococcus aureus ATCC 27853, Escherichia coli ATCC 25922 และ Pseudomonas aeruginosa แอคติโนแบคทีเรียทั้งหมด 40 ไอโซเลต แยกได้จากดินบริเวณรอบรากฟ้าทะลายโจร จำนวน 17 ไอโซเลต ดินบริเวณรอบรากกระท่อม จำนวน 12 ไอโซเลต และดินบริเวณรอบรากกัญชา จำนวน 11 ไอโซเลต ในจำนวนนี้มี 11 ไอโซเลต ยับยั้งแบคทีเรียทดสอบได้อย่างน้อย 1 ชนิด และมี 3 ไอโซเลต คือ KT01, KC11 และ F17 มีกิจกรรมการยับยั้งแบคทีเรียดีที่สุดและคัดเลือกมาทดสอบการยับยั้งการสร้างไบโอฟิล์มด้วยวิธี microtiter plate (MTP) assay ผลการศึกษาพบว่าไอโซเลต KT01 มีกิจกรรมยับยั้งการสร้างไบโอฟิล์มดีที่สุด น้ำเลี้ยงเชื้อปราศจากเซลล์ของไอโซเลต KT01 ยับยั้งการสร้างไบโอฟิล์มของ B. cereus TISTR 687, S. aureus ATCC 27853, E. coli ATCC 25922 และ P. aeruginosa ได้ 11.69 ± 0.06%, 19.32 ± 0.05%, 23.57± 0.37% และ 87.46 ± 0.07% ตามลำดับ จากการวิเคราะห์ลำดับนิวคลีโอไทด์ของยีน 16S rDNA ของไอโซเลต KT01 (1,499 คู่เบส) พบว่ามีความเหมือนกัน กับ Streptomyces griseoincarnatus LMG 19316T (100 เปอร์เซ็นต์) จากผลการศึกษานี้แสดงให้เห็นว่าแอคติโนแบคทีเรียที่แยกจากดินรอบรากพืชสมุนไพรเป็นแหล่งของสารออกฤทธิ์ทางชีวภาพที่มีฤทธิ์ยับยั้งแบคทีเรียสาเหตุโรคและยับยั้งการสร้างไบโอฟิล์มได้
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