Phân tích thành phần quần xã vi sinh vật trong các mô hình xử lý AMD trong phòng thí nghiệm

Để theo dõi thành phần của quần xã vi khuẩn trong các mô hình xử lý, đặc biệt là các vi khuẩn khử sulfate, chúng tôi tiến hành phân tích đoạn gen 16S rDNA bằng phương pháp điện di biến tính (DGGE) (hình 3.14).

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SR2

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Trước hết có thể thấy rằng ba chủng SRB phân lập được (đường điện di số 4, 5, 6) là đại diện của các nhóm vi khuẩn chiếm số đông trong mẫu làm giàu E1-4 (đường điện di số 1, các băng có mũi tên chỉ).

Trong quá trình vận hành mô hình xử lý với methanol là chất hữu cơ được bổ sung làm cơ chất cho SRB, cả 3 nhóm SRB mà các chủng SR2, SR3 và SR4 làm đại diện đều không được duy trì ở mật độ cao, thay vào đó là nhiều nhóm vi khuẩn khác phát triển, đặc biệt nhóm vi khuẩn ở băng đánh dấu * (đường điện di số 2).

Khi chất thải hữu cơ được bổ sung làm cơ chất cho SRB trong bể xử lý AMD, SR4 là đại diện của một trong các nhóm chính được tìm thấy (băng có mũi tên chỉ, đường điện di số 3). Bên cạnh đó hai nhóm khác ở các băng đánh dấu * và ** cũng là các nhóm chính nhưng chưa được xác định (đường điện di số 3).

1. 
   Đã thiết lập được hỗn hợp SRB (mẫu E1-4) có hoạt tính tốt trong điều kiện pH thấp qua phương pháp làm giàu.
   
2. 
   Phân lập được 3 chủng vi khuẩn khử sulfate SR2, SR3 và SR4 từ mẫu làm giàu nói trên. Dựa trên trình tự gần đủ của gen 16S rDNA các chủng này được định danh tương ứng là Desulfomicrobium sp. SR2, Desulfobulbus sp. SR3, và Desulfovibrio sp. SR4.
   
3. 
   Phân tích DGGE gen 16S rDNA cho thấy các chủng phân lập đại diện cho các nhóm SRB chính trong mẫu làm giàu E1-4.
   
4. 
   Nghiên cứu đặc điểm sinh lý của 3 chủng thấy rằng:
   

• 
  Cả 3 chủng đều có khả năng sinh trưởng tốt ở môi trường có hàm lượng muối 10 – 15 g/l, tương ứng với môi trường nước lợ. Đặc biệt là chủng SR4 có thể sinh trưởng tốt tại nồng độ muối 25 g/L, tương đương môi trường nước biển.
  
• 
  Cả ba chủng đều bị ức chế tại pH môi trường  6, tuy nhiên mẫu làm giàu gốc E1-4 thể hiện khả năng chịu pH thấp tốt hơn các chủng thuần khiết và sinh trưởng tốt ở pH 4 và 5.
  
• 
  Ba chủng đều sinh trưởng và tạo sulfide tối ưu ở 30^oC và được xếp vào nhóm SRB ưa ấm điển hình.
  
• 
  Chất cho điện tử được cả 3 chủng sử dụng hiệu quả nhất là lactate. Chủng SR2 và SR3 còn sử dụng acetate và methanol nhưng với hiệu quả thấp hơn, trong khi đó SR4 hoàn toàn không sử dụng acetate.
  
• 
  Ngoài chất nhận điện tử chính là sulfate, 2 chủng SR2 và SR4 còn sử dụng được cả nitrate làm chất nhận điện tử. Ngoài ra, chủng SR4 còn oxy hóa được lactate bằng Fe³⁺.
  

5. 
   Mô hình thử nghiệm xử lý AMD với cơ chất bổ sung là nước thải có hàm lượng hữu cơ cao đạt hiệu quả cao hơn khi sử dụng cơ chất đơn là methanol. Kết quả thu được sau 7 ngày xử lý gồm: pH tăng từ 4 lên 8,15, nồng độ sulfate giảm từ 13,75 mM còn 4,5 mM, hàm lượng sắt giảm từ 200 mg/l còn 36 mg/l thể hiện khả năng ứng dụng thực tế của công nghệ.
   
6. 
   Phân tích điện di biến tính gen 16S rDNA cho thấy Desulfovibrio spp., đại diện là chủng SR4 đóng vai trò quan trọng trong bước xử lý AMD với nguồn cơ chất bổ sung là nước thải có hàm lượng hữu cơ cao.
   

1. 
   Xác định các nhóm vi khuẩn khác SR4 cũng đóng vai trò quan trọng trong quá trình xử lý AMD ở MH2.
   
2. 
   Phân lập thêm các chủng SRB ưa axit để bổ sung vào mô hình xử lý AMD.
   
3. 
   Thử nghiệm áp dụng công nghệ ở quy mô lớn hơn trong điều kiện thực tế.
   

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GGTATCCGAGCGACTTCGGGTAGAACCGACTCTCGTGGTGTGAACGGGCGGTGTGTCACAAGGCCCGGGAACGTATTCACCCCGGCATGCTGATCCGGCGATTACTGAGCGATTCCAACTTCAGGCAGGCGAGTCGAAGACTGCAATCCGGACTATGGATGGGTTTTTTGAGATTCGCTCGACCTCACGGTTTCGCTGCCCTTTGTACCCACCATTGTAATACGTGTGGTAGCCCTAGGCGTAAGGGCCATGATGACTTGAAGTCATCCCCACCTTCTCTCCCGGTTAACGCGGGCAGTCTCCACAGAGTGCCCACCATTATGTGCTGGCAACTGAGAATAGGGGTTGCGCTCGTTGCGGGACTTAACCCAACACCTCACGGCACGAGCTGACGACAGCCATGCAGCACCGGTCTCTGGATTCCCCGAAGGGCACTCCCGCATCTCTGCAGGATTCCCAGGATGTCAAGCCTAGGTAAGGTTCTTCGCGTTGCATCGAAATAAACCACCATACTCCACCGCTTGTGCGGGCCCCCGTCAATTTCTTTGAGTTTCAGCCTTGCGACCGTACTCCCCAGGCGGGATACTTAACGCGTTAGCTACGGCACCGAAGATCAAGTCCCCGACACCTAGTATCCATCGTTTACGGTGTGGACTACCACGGTATCTAATCCTGTTTGCTCCCCACACTTTCGCACCTCAACGTCAATACCTGTCCAGGTGGCCGCCTTCGCCACCGGTGTTCCTCCTGATATCTACGGATTTCACTCCTACACCAGGAATTCCGCCACCCTCTCCAGGATTCAAGCCCTGCAGTTTCAAAGGCAGTTCCACGGTTGAGCCGTGGGATTTCACCCCTGACTTACAAGGCCGCCTACGTGCGCTTTACGCCCAGTAATTCCGAATAACGCTTGCACCCTCCGTATTACCGCGGCTGCTGGCACGGAGTTAGCCGGTGCTTCCTCTAAAGGTACCGTCAAAACAAAGGCCTATTACACCAATGCCCGTTCTTCCCTTCCGACATGAGGTTTACGACCCGAAAGCCTTCATCCCTCACACGGCGTTGCTGCGTCAGGCTTTCGCCCATTGCGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGACCGTGTTTCAGTTCCAGTGTGGCTGATCATCCTCTCAGACCAGCTACTCATCGTTGCCTTGGTAGGCCATTACCCTACCAACTAGCTAATGAGACGCGGGCTCATCCTCGGACGAATGCATATGCAGAGGCATCCTTTACCGACGTCTATTAAAAGCCAGACTATCCGGTATTAGCTCCACTTTCGCGGAGTTATTCCAAATCCAAGGGTAGATTACCCACGCGTTACTCACCCGTGCGCCACTCTACTCAGGGCCGAAGCCCCTTTCTCGTACGACTGCAAGGTGTTAGGAACGCCGCCGGCGTCTCCCATTCTGA

GCGCATCGCAGGCGGCGATGATTATCACATGCAAATCGAACGCGAAAGGGACTTCGGTCCTGAGTAAAGTGGCGCAAGGGTGACTATTACGTGTAGATAACCCGTCTTCATGTATGGAATAATACACCGAAAGGGGTACTAATACCGGATATTATTGCCTCATATAAGTTTTGCAAGCAAAGGTGGCATCTGATTCAAGCTACTGCATGTAGAGGCGTCTGCGTACCATGAGCTAGTAGGTAGGGTAATGGCCTACCTAGGCAACGATGGTGAGCGGGTCTGAGAGGATGATCCGCCACACTGGCACTGGAACACGGGCCAGACTCCTACGGGAGGCAGCAGTGAGGAATATTGCGCAATGGGGGCAACCCTGACGAAGCGACGCCGCGTGAGTGAGGAAGGCCTTCGGGTCGTAAAGATCTGTCAAGAGGAAAGAAATGCATAGCGGTTAATACCCGCTATGTTTGACGGTACCTCTAAAGGAAGCACAGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTGTTCGGAATCACTGGGCGTAAAGGGCGCGCAGGCGGTTTGGTAAGTCAGATGTGAAAGCCCACGGCTTAACCGTGGAAGTGCATTTGATACTGCTAGACCTGAGTACCAGAGGGGAAAGTGGAATTCCCGGTGTAGAGGTGAAATTCGTAGATATCGGGAGGAATACCGGTGGCGAAGGCGACTTTCTGGCTGAGTACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGATGTAGGGGGGTGTTGATCCATTCTGTGTCGCAGCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGTATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGTCTTGACATCCCGGGAATCTTTCGGAAACGAGAGAGTGCCTTCATTAGAAGGAACCTGGAGACAGGTGCTGCATGGCAGTCGTCAGCTCGTGTCATGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGCCTTTAGTTGCCAGCAGTTCGGCTGGGCACTCTAAAGGGACGGCCGGTGTCAAACCGAAGAAAGGTGGGGATGACGTCAAGTCCTCATGGCCTTTATGACCAGGGCTACACACGTACTTACAATGGCCGATACAAAGGGCAGGCCACATTGCGAAATGGAGCCAATCCCATAAAACCGCTCTCACTCCGAATTGAAGTCTGCAACTCGACTCCATGGAAGTTGGAATCCCTAGTAATCGCGGATCAGCAATGCCGCGGGGAAAACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACCGGAATCGGTTGTACCAAAAGCAGTA

GTTACCCCGACAGTTTTAGGTAGGAACCGACTTTCGTGGTGTAGACGGGCGGTGTGTAACAAGGCCCGGGAAAGTATTCACCCGGGACCATGCTGATCTCCGATATACAAGCGATTCCGACTTCACGGGGGTCGAATTGCAGACCCCTGATCCGGAATGGGACCGGTTTTTGGGGATTGGCTCCACCCTGCGGTCTCGCAAGCCCATTGTAACCCGCCATTGTAGTACGTGTGTAGCCCTGGGCGTAAGGCCCATGATGACTTGACGTCGTCCCCACCTTCCTTCCCCGTTGACCGAGGCGGTCTCCCTAGAGTGCCCGACATTACTCGCTGGCAACTAAGGACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACCACCTCACGGCACGAGCTGACGACAGCCATGCAGCACCTGTCACCCCGCTCCCCGAAGGGCACTCCTCCTTTTCGGGAGGATTCGAGGGATGTCAAACCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAAACCACATACTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGCCTTGCGACCGTACTCCCCAGGCGGGATGCTTAATGCGTTAACTGCGGCACCGAAGATCGCTCCCCGACACCTAGCATCCATCGTTTACAGCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGCGCCTCAGCGTCAGTACCTGTCCAGGTGGCCGCCTTCGCCACCGGTGTTCCTCCGGATATCTACGGATTTCACTCCTACACCCGGAATTCCGCCACCCTCTCCAGGACTCAAGTCTCCCAGTATCGAACGCAGTTCCCCGGTTGAGCCGAGGGCTTTCACGTCCGACTTAAAAGACGGCCTACGCGCGCTTTACGCCCAGTGATTCCGATTAACGCTTGCACCCTCCGTATTACCGCGGCTGCTGGCACGGAGTTAGCCGGTGCTTCCTCTGGAAGTACCGTCAGTCCCAAGGCTTGTTCAGCCTTGAGAGGTTCTTCCTTCCTGACAGAGGTTTACGACCCGAAAGCCTTCTTCCCTCACACGGCGTCGCTGCGTCAGGGTTTCCCCCATTGCGCAATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGACCGTGTTTCAGTTCCAGTGTGGCTGATCATCCTCTCAGACCAGCTACCCATCGTTGCCTTGGTGGGCCATTACCCCACCAACAAGCTAATGGGACGCGGACTCATCTCATTGCGACAGCTTGCAAGCAGAGGCCGCCTTTCCCCCAACCGAAATTGGAGCGTATCCGGTATTAGCGGCAGTTTCCCGCCGTTATCCCAAACAATGAGGGAGATAATCCACGCGTTACTCACCCGTGCGCCGCTCTACTCAGGGACCGAAGCCCCCTTTCTCGCACGACTTGCACGTGTTAAGCACGCCCCCACCGTTGATCTG