Name: laboratory simulation of an iron(ii)-rich precambrian marine upwelling system to explore the growth of photosynthetic bacteria
Uploaded: 2016-07-24T15:00:00.000+00:00
Duration: 9 min 45 s
Description: Watch this Scientific Journal Video about Laboratory Simulation of an IronII-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria at JoVE.com

마크 노프 호프는 배관 연결의 설계 및 구현에 도움. 엘렌 STRUVE 선택하고 사용하는 장비를 획득 할 수있었습니다.

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저자는 공개 아무것도 없어.

선캄브리아 바다에서 미생물 커뮤니티에 의해 규제, 또는 활동의 결과 및 일반적인 화학적 조건으로 수정되었습니다. BIF의 기원을 해석에서 연구자들은 일반적으로의 퇴적 또는 BIF의 지구 화학, 예를 들면, 스미스 등. (23)와 존슨 등. (24)에 따라 미생물의 존재 활동을 추론. 고대의 환경 지구 화학적 유사체가 현대적인 환경에서 현대 생물의 연구는 가치있는 ?...

선캄브리아는 (4.6 Gy를 0.541으로 전) 분위기가 광합성 생산 된 산소의 점진적 형성을 경험 (O 2), 아마도 약 2.4 Gy의에서 이른바 &quot;위대한 산화 이벤트&quot;(GOE)에서 단계 변화에 의해 구두점 전 및 다시 대기 O와 신 원생대 (1-0.541 Gy를 전) 2 현대 레벨 1에 접근했다. 시아 노 박테리아는 산소를 광합성이 할 수있는 첫 번째 생물의 진화 잔재입니다. 지구 화학적 증거 및 모델링 연구는 주로 무산소 분위기 3-5 아래 표면 바다에서 로컬 산소 오아시스를 생성, 시아 노 박테리아 또는 산소를 광합성 또는 산소를 광 영양 생물의 수 생물의 활동 지역 사회를 품고 얕은 연안 환경의 역할을 지원합니다. 철 선캄브리아 점 걸쳐 해수 줄무늬 철 구조물 증착 (BIFS) (II) (철 (II))의 주요 화학적 C로서이들 증착 동안 적어도 국부적 해수 onstituent,. 가장 큰 BIFS 중 일부는 대륙붕 및 사면을 형성, 깊은 물 예금이다. 증착 철의 양은 주로 대륙 (즉, 풍화) 소스와 질량 균형의 관점에서 호환되지 않습니다. 따라서 철의 많은에 mafic 나에 ultramafic 해저 지각 (6)의 열수 변질로부터 공급되어 있어야합니다. 철의 비율의 추정은 연안 환경의 아웃 보드가 용승 (7)을 통해 표면 바다에 공급 철 (II)와 일치 증착. 철은 용승 전류에 전송 받으려면 감소, 모바일 형태로 존재되어 있어야합니다 - 철과 같은 (II). BIF 보존 철의 평균 산화수는 2.4 (8)이며, 일반적으로는 철 BIF는 산소 가능성, (II)이 산화 된 철을 승이시 형성된 철 (III), 퇴적 유지한다고 생각된다. 따라서, 경사 environme 따라 전위의 Fe (II) 산화 메커니즘을 탐구국세청은 BIF 형성 방법을 이해하는 것이 중요합니다. 또한, 해양 퇴적물의 세련된 지구 화학적 특성은 철 (Ⅱ) 무산소 물기둥의 존재 철분을 함유 조건, 선캄브리아에 걸쳐 바다의 지속적인 기능이었고, 단지 시간과 장소에 제한되지 않을 수 있음을 확인했습니다 여기서 BIF 9 증착 하였다. 따라서, 지구 역사의 적어도 이십억년를 들어, 얕은 바다에서의 Fe (II)와 O (2) 사이의 산화 환원 인터페이스 가능성이 평범했다. 많은 연구는 캄브리아기 바다의 다양한 기능의 화학적 및 / 또는 생물학적 유사체있는 현대적인 사이트를 사용합니다. 좋은 예는 광합성 활동 (시아 노 박테리아에 의해 포함) 동안의 Fe (II)는 10 ~ 13을 검출 하였다 햇볕에 쬐 표면 바다에서 안정적으로 존재하는 철분을 함유 호수입니다. 이 연구의 결과는 / FER을 무산소 할 수있는 호기의 지구 화학적 및 미생물 특성에 대한 통찰력을 제공ruginous chemocline. 그러나이 위치는 일반적으로 물리적으로 거의 수직 오히려 승이 시스템에서 발생하는 화학 인터페이스보다 14 믹싱 층화하고, 4 시간 선캄브리아 가장 산소 생성을 지원하는 것으로 생각된다. 자연 아날로그는 무산소 분위기 아래 해양 산소 오아시스의 발전을 탐구하고, 햇볕에 쬐 지표수 열에서의 Fe (II)이 풍부한 용승 시스템에서 현대 지구에서 사용할 수 없습니다합니다. 따라서, 함철 승이 영역을 시뮬레이션하며 박테리아 및 photoferrotrophs의 성장을 지원할 수있는 실험실 시스템이 필요하다. 선캄브리아 해수를 나타내는 이해와 미생물 공정의 식별과 용승 수성 매질과의 상호 작용에 대한 이해를 촉진하고 완전히 고대 지구에 특유의 생지 화학적 과정을 이해하기 위해 암석 기록에서 얻은 정보를 보완 할 수 있습니다. </p&gt; 이를 위해 실험실 규모 컬럼되는 철 (II)이 풍부한 해수 배지 (중성)이 컬럼의 바닥으로 펌핑 설계하고, 상부에서 펌핑. 조명은 최고 3cm의 시아 노 박테리아의 성장을지지 4cm 폭 &quot;빛의 영역&quot;을 만들기 위해 상단에 제공되었다. 자연 환​​경은 일반적으로 층화 및 염분이나 온도 등의 물리 화학적 구배에 의해 안정화된다. 실험실 규모의 수층을 안정화시키기 위해, 열 실린더는 실험 기간 동안 개발 된 화학적 패턴의 설정을 유지하는 데 도움이 다공질 유리 비드 매트릭스 채워졌다. 연속적인 N 2 / CO 2 가스 유동은 GOE 15 전에 바다 반사 무산소 분위기를 유지하기 위해 컬럼의 상부 공간을 세척하기 위해 적용 하였다. 철의 일정한 플럭스 (II)를 설치 한 후, 시아 노 박테리아는 열에 걸쳐 접종 및 growt했다H는 샘플링 포트를 통해 제거 된 샘플을 세포 수로 모니터링 하였다. 산소는 열 실린더 측정 내벽 열 외부 광섬유로 만들어진 상 산소 민감성 optode 박을 배치하여 현장에서 모니터링 하였다. 수성 철의 분화는 깊이 해결 수평 샘플링 포트에서 제거 샘플에 의해 정량화하고 Ferrozine 방법으로 분석 하였다. 비 생물 제어 실험 결과는 개념 증명을 보여 - 대기로부터 격리 유지 고대의 물기둥의 실험실 규모의 아날로그, 달성이다. 시아 노 박테리아가 성장하여 산소를 생산하고, 철 (II)와 산소의 반응을 확인할 수 있었다. 해양 cyanobacterium 인 Synechococcus 특검팀 접종 동안 여기서, 디자인, 제조, 조립, 실행, 이러한 열의 샘플링의 방법은 컬럼의 84 시간 실행에서 결과와 함께 제공됩니다. PCC (7002).

<table><tbody><tr><td>Widdel flask (5 L)</td><td>Ochs</td><td>110015</td><td>labor-ochs.de</td></tr><tr><td>Glass bottles (5 L)</td><td>Rotilabo</td><td>Y682.1</td><td>carlroth.com</td></tr><tr><td>Glass pipettes (5 ml)</td><td></td><td>51714</td><td>labor-ochs.de</td></tr><tr><td>0.22 &amp;micro;m Steritop filter unit (0.22&amp;nbsp;&amp;micro;m Polyethersulfone membrane)</td><td>Millipore</td><td>X337.1</td><td>carlroth.com</td></tr><tr><td>Aluminum foil</td><td></td><td></td><td></td></tr><tr><td>Sterile Luer Lock glass syringe, filled with cotton</td><td></td><td>C681.1</td><td>carlroth.com</td></tr><tr><td>Luer Lock stainless steel needles (150 mm, 1.0&amp;nbsp;mm ID)</td><td></td><td>201015</td><td>labor-ochs.de</td></tr><tr><td>NaCl</td><td>Sigma</td><td>433209</td><td>sigmaaldrich.com</td></tr><tr><td>MgSO&lt;sub&gt;4&lt;/sub&gt;</td><td>Sigma</td><td>208094</td><td>sigmaaldrich.com</td></tr><tr><td>CaCl&lt;sub&gt;2&lt;/sub&gt;</td><td>Sigma</td><td>C4901</td><td>sigmaaldrich.com</td></tr><tr><td>NH&lt;sub&gt;4&lt;/sub&gt;Cl</td><td>Sigma</td><td>A9434</td><td>sigmaaldrich.com</td></tr><tr><td>KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;</td><td>Sigma</td><td>P5655</td><td>sigmaaldrich.com</td></tr><tr><td>KBr</td><td>Sigma</td><td>P3691</td><td>sigmaaldrich.com</td></tr><tr><td>KCl</td><td>Sigma</td><td>P9541</td><td>sigmaaldrich.com</td></tr><tr><td>Glass cylinder</td><td></td><td>Y310.1</td><td>carlroth.com</td></tr><tr><td>Glass wool</td><td></td><td>7377.2</td><td>carlroth.com</td></tr><tr><td>Glass beads (&amp;oslash; 0.55 - 0.7&amp;nbsp;mm)</td><td></td><td>11079105</td><td>biospec.com</td></tr><tr><td>Butyl rubber stopper (&amp;oslash; 1.2 cm)</td><td></td><td>271024</td><td>labor-ochs.de</td></tr><tr><td>Petri Dish, glass (&amp;oslash; 8.0&amp;nbsp;cm)</td><td></td><td>T939.1</td><td>carlroth.com</td></tr><tr><td>Polymers glue</td><td></td><td>OTTOSEAL S68</td><td>adchem.de</td></tr><tr><td>Optical oxygen sensor foil (for oxygen analysis, see below)</td><td></td><td>&amp;ndash; on request &amp;ndash;</td><td>presens.de</td></tr><tr><td>Rubber tubing (35 mm, 7&amp;nbsp;mm ID)</td><td></td><td>770350</td><td>labor-ochs.de</td></tr><tr><td>Luer Lock tube connector (3.0 mm, Luer lock male = LLM)</td><td></td><td>P343.1</td><td>carlroth.com</td></tr><tr><td>Luer Lock tube connector (3.0 mm, Luer lock female = LLF)</td><td></td><td>P335.1</td><td>carlroth.com</td></tr><tr><td>Rubber tubing (25 mm, 0.72 mm ID)</td><td></td><td>2600185</td><td>newageindustries.com</td></tr><tr><td>Rubber tubing (50 mm, 7&amp;nbsp;mm ID)</td><td></td><td>770350</td><td>labor-ochs.de</td></tr><tr><td>Luer Lock stainless steel needle (150 mm, 1.0 mm ID)</td><td></td><td>201015</td><td>labor-ochs.de</td></tr><tr><td>Luer Lock glass syringe (10 ml)</td><td></td><td>C680.1</td><td>carlroth.com</td></tr><tr><td>Loose cotton&amp;nbsp;</td><td></td><td>&amp;ndash;</td><td></td></tr><tr><td>Butyl rubber stopper (&amp;oslash; 1.75 cm)</td><td></td><td>271050</td><td>labor-ochs.de</td></tr><tr><td>Stainless steel needle (40 mm, 1.0&amp;nbsp;mm ID)</td><td>Sterican</td><td>4665120</td><td>bbraun.de</td></tr><tr><td>Luer Lock stainless steel needle (150 mm, 1.5 mm ID)</td><td></td><td>201520</td><td>labor-ochs.de</td></tr><tr><td>position: Luer Lock female connector part at C.7</td><td></td><td></td><td></td></tr><tr><td>Polymers glue</td><td></td><td>OTTOSEAL S68</td><td>adchem.de</td></tr><tr><td>Stainless steel needle (120 mm, 0.7 mm ID)</td><td>Sterican</td><td>4665643</td><td>bbraun.de</td></tr><tr><td>Rubber tubing (40 mm, 0.74 mm ID)</td><td></td><td>2600185</td><td>newageindustries.com</td></tr><tr><td>Heat shrink tubing (35&amp;nbsp;mm, 3 mm ID shrunk)</td><td></td><td>541458 - 62</td><td>conrad.de</td></tr><tr><td>Tube clamp</td><td></td><td>STHC-C-500-4</td><td>tekproducts.com</td></tr><tr><td>Luer Lock tube connector (1.0 mm, LLF)</td><td></td><td>P334.1</td><td>carlroth.com</td></tr><tr><td>Luer Lock plastic cap (LLM)</td><td></td><td>CT69.1</td><td>carlroth.com</td></tr><tr><td>Glass bottle (5 L)</td><td>Rotilabo</td><td>Y682.1</td><td>carlroth.com</td></tr><tr><td>Butyl rubber stopper (for GL45)</td><td></td><td>444704</td><td>labor-ochs.de</td></tr><tr><td>Stainless steel capillary (300 mm, 0.74 mm ID)</td><td></td><td>56736</td><td>sigmaaldrich.com</td></tr><tr><td>Stainless steel capillary (50 mm, 0.74 mm ID)</td><td></td><td>56737</td><td>sigmaaldrich.com</td></tr><tr><td>Shrink tubing (35 mm, 3&amp;nbsp;mm ID shrunk)</td><td></td><td>541458 - 62</td><td>conrad.de</td></tr><tr><td>Rubber tubing (100 mm, 0.74 mm ID)</td><td></td><td>2600185</td><td>newageindustries.com</td></tr><tr><td>Luer Lock tube connector (1.0 mm, LLF)</td><td></td><td>P334.1</td><td>carlroth.com</td></tr><tr><td>Luer Lock glass syringe (10 ml)</td><td></td><td>C680.1</td><td>carlroth.com</td></tr><tr><td>Loose cotton&amp;nbsp;</td><td></td><td>&amp;ndash;</td><td></td></tr><tr><td>Butyl rubber stopper (&amp;oslash; 1.75 cm)</td><td></td><td>271050</td><td>labor-ochs.de</td></tr><tr><td>Stainless Steel needle (40 mm, 0.8&amp;nbsp;mm ID)</td><td>Sterican</td><td>4657519</td><td>bbraun.de</td></tr><tr><td>Luer lock glass syringe (5 ml)</td><td></td><td>C679.1</td><td>carlroth.com</td></tr><tr><td>Butyl rubber stopper (&amp;oslash; 1.75 mm)</td><td></td><td>271050</td><td>labor-ochs.de</td></tr><tr><td>Rubber tubing (40 mm, 0.74 mm ID)</td><td></td><td>2600185</td><td>newageindustries.com</td></tr><tr><td>Glass bottle (2 L)</td><td>Rotilabo</td><td>X716.1</td><td>carlroth.com</td></tr><tr><td>Butyl rubber stopper (for GL45)</td><td></td><td>444704</td><td>labor-ochs.de</td></tr><tr><td>Stainless steel capillary (50 mm, 0.74 mm ID)</td><td></td><td>56736</td><td>sigmaaldrich.com</td></tr><tr><td>Rubber tubing (30 mm x 0.74 mm ID)</td><td></td><td>2600185</td><td>newageindustries.com</td></tr><tr><td>Rubber tubing (100 mm x 0.74 mm ID)</td><td></td><td>2600185</td><td>newageindustries.com</td></tr><tr><td>Luer Lock tube connector (1.0 mm, LLF)</td><td></td><td>P334.1</td><td>carlroth.com</td></tr><tr><td>Luer Lock 3-way connector (LLF, 2x LLM)</td><td></td><td>6134</td><td>cadenceinc.com</td></tr><tr><td>Light source</td><td>Samsung</td><td>SI-P8V151DB1US</td><td>samsung.com</td></tr><tr><td>Peristalic pump</td><td>Ismatec</td><td>EW-78017-35</td><td>coleparmer.com</td></tr><tr><td>Pumping tubing (0.89&amp;nbsp;mm ID)</td><td></td><td>EW-97628-26</td><td>coleparmer.com</td></tr><tr><td>Stainless steel capillary (200 mm, 0.74 mm ID)</td><td></td><td>56736</td><td>sigmaaldrich.com</td></tr><tr><td>Stainless steel capillary (400 mm, 0.74 mm ID)</td><td></td><td>56737</td><td>sigmaaldrich.com</td></tr><tr><td>Supel-Inert Foil (Tedlar - PFC) gas pack (10 L)</td><td></td><td>30240-U</td><td>sigmaaldrich.com</td></tr><tr><td>Rubber tube (30 mm, 6&amp;nbsp;mm ID)</td><td></td><td>770300</td><td>labor-ochs.de</td></tr><tr><td>Luer Lock tube connector (3.0 mm, LLM)</td><td></td><td>P343.1</td><td>carlroth.com</td></tr><tr><td>Luer Lock tube connector (3.0 mm, LLF)</td><td></td><td>P335.1</td><td>carlroth.com</td></tr><tr><td>Gas-tight syringe (20&amp;nbsp;ml)</td><td></td><td>C681.1</td><td>carlroth.com</td></tr><tr><td>Bunsen burner</td><td></td><td>&amp;ndash;</td><td></td></tr><tr><td>Fiber optic oxygen meter for oxygen quantification</td><td>Presens</td><td>TR-FB-10-01</td><td>presens.de</td></tr><tr><td>Vacuum pump</td><td></td><td>&amp;ndash;</td><td></td></tr><tr><td>Silicone glue for oxygen optodes</td><td>Presens</td><td>PS1</td><td>presens.de</td></tr></tbody></table>

laboratory simulation of an iron(ii)-rich precambrian marine upwelling system to explore the growth of photosynthetic bacteria

일부 선캄브리아 줄무늬 철 구조물 (BIF)을 증착하기위한 종래의 개념 선캄브리아 바다 열수 출처 1 철 [철 (II)] 승이은 분자 산소에 의해 산화 된 것으로 가정하여 진행한다 O 2] 박테리아에 의해 생산. 전 약 24억년 (Gy를)에서 이전에 좋은 산화 이벤트 (GOE)에 증착 된 가장 오래된 BIFS는, 무산소 조건에서 anoxygenic photoferrotrophs에 의해 철 (II)의 직접 산화에 의해 형성 할 수 있었다. 다른 생물학적 시나리오에 따라 개발하는 지구 화학적 및 광물 학적 패턴을 테스트하기위한 방법으로, 우리는 실험실 규모의이 풍부한 해양 용승 시스템 고대 바다의 대표적인 무산소의 Fe (II)을 시뮬레이션 할 수있는 40cm 길이의 수직 흐름을 통해 열 설계 . 실린더는 화학적 구배를 안정화 다공질 유리 비드 매트릭스로 포장하고, 철 정량 액체 시료 수층에 걸쳐 수행 될 수있다. 용존 산소이었다외부에서 optodes를 통해 비 침습적 발견했습니다. 물 열에서 하단, 상단에서 별개의 빛 그라데이션 및 시아 노 박테리아의 존재에서의 Fe (II)의 용승 플럭스를 포함 생물 실험, 철의 형성을위한 쇼 명백한 증거의 결과 (III) 미네랄 침전물과 chemocline 개발 철 사이 (II) 및 O 2. 이 열은 우리가 시뮬레이션 해양 선캄브리아 조건 (미래 photoferrotrophs에서) 시아 노 박테리아를 배양하여 BIFS의 형성 가설을 테스트 할 수 있습니다. 얕은 바다 또는 호수의 퇴적물을 포함하여 - 또한 우리는 우리의 열 개념은 다양한 화학적, 물리적 환경의 시뮬레이션 수 있다는 가설.

제어 실험 비 생물 제어 실험 (십일)이 지속적으로 낮은 산소 농도를 보여 주었다 (O 2 &lt;유한 요소 (II에서 유의 한 변동와 0.15 ㎎ / ℓ)) -profile 위로 향하는 물기둥을 통해. 석출물의 형성은 (아마도의 Fe (III) (oxyhydr-)의 산화물) 배지 저장조와 연결을 통해 10 일 일부 산소 확산을 나타내는 500 μM 내...

Watch this Scientific Journal Video about Laboratory Simulation of an IronII-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria at JoVE.com

Laboratory Simulation of an IronII-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

우리는 실험실 규모의 수직 흐름을 통해 열에서 선캄브리아 철분을 함유 해양 용승 시스템을 시뮬레이션. 목표는 O 2 및 Fe (II)의 방법 화학적 프로파일을 이해 시아 노 박테리아의 생산 O 2로 진화했다. 이 결과로 인해 광합성 제조 O 2에 의해 철 (II) 산화에 chemocline의 설립을 나타낸다.

선캄브리아 해양 용승 시스템 광합성 박테리아의 성장을 탐구하는이 풍부한 철 (II)의 실험실 시뮬레이션

A conventional concept for the deposition of some Precambrian Banded Iron Formations (BIF) proceeds on the assumption that ferrous iron [Fe(II)] upwelling ...

laboratory-simulation-an-iron-ii-rich-precambrian-marine-upwelling

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JoVE Journal

Environment

Laboratory Simulation of an Iron(II)-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

11.7K 조회수.  University of Tuebingen.  우리는 실험실 규모의 수직 흐름을 통해 열에서 선캄브리아 철분을 함유 해양 용승 시스템을 시뮬레이션. 목표는 O 2 및 Fe (II)의 방법 화학적 프로파일을 이해 시아 노 박테리아의 생산 O 2로 진화했다. 이 결과로 인해 광합성 제조 O 2에 의해 철 (II) 산화에 chemocline의 설립을 나타낸다. 

비디오: Laboratory Simulation of an IronII-rich Precambrian Marine Upwelling System to Explore the Growth of Photosynthetic Bacteria

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated (WATER-PAM) Fluorometry

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be subsampled throughout the experiment1&#8211;7. Subsampling of large volumes can be problematic for several reasons: 1) it causes variation in the total volume and the surface area:volume ratio of the culture during the experiment; 2) pseudo-replication (i.e., replicate samples from the same treatment flask8) is often employed rather than true replicates (i.e., sampling from replicate treatments); 3) the duration of the experiment is limited by the total volume; and 4) axenic cultures or the usual bacterial microbiota are difficult to maintain during long-term experiments as contamination commonly occurs during subsampling.
The use of microtiter plates enables 1 ml culture volumes to be used for each replicate, with up to 48 separate treatments within a 12.65 x 8.5 x 2.2 cm plate, thereby decreasing the experimental volume and allowing for extensive replication without subsampling any treatment. Additionally, this technique can be modified to fit a variety of experimental formats including: bacterial-algal co-cultures, algal physiology tests, and toxin screening9&#8211;11. Individual wells with an alga, bacterium and/or co-cultures can be sampled for numerous laboratory procedures including, but not limited to: WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry, microscopy, bacterial colony forming unit (cfu) counts and flow cytometry. The combination of the microtiter plate format and WATER-PAM fluorometry allows for multiple rapid measurements of photochemical yield and other photochemical parameters with low variability between samples, high reproducibility and avoids the many pitfalls of subsampling a carboy or conical flask over the course of an experiment.

A Small Volume Bioassay to Assess Bacterial/Phytoplankton Co-culture Using WATER-Pulse-Amplitude-Modulated WATER-PAM Fluorometry

The goal of this procedure is to demonstrate the reproducibility and adaptability of using a microtiter plate format for microalgal screening. This rapid screen combines WATER-Pulse-Amplitude-Modulated (WATER-PAM) fluorometry to measure photosynthetic yield as an indicator of Photosystem II (PSII) health with small volume bacterial-algal co-cultures.

작은 볼륨 생물 검정은 물 - 펄스 진폭 변조 (WATER-PAM) 형광 측정을 사용하여 세균 / 식물 플랑크톤의 공동 문화를 평가하는

Conventional methods for experimental manipulation of microalgae have employed large volumes of culture (20 ml to 5 L), so that the culture can be ...

연구

환경과학

Quantification of Heavy Metals and Other Inorganic Contaminants on the Productivity of Microalgae

Increasing demand for renewable fuels has researchers investigating the feasibility of alternative feedstocks, such as microalgae. Inherent advantages include high potential yield, use of non-arable land and integration with waste streams. The nutrient requirements of a large-scale microalgae production system will require the coupling of cultivation systems with industrial waste resources, such as carbon dioxide from flue gas and nutrients from wastewater. Inorganic contaminants present in these wastes can potentially lead to bioaccumulation in microalgal biomass negatively impact productivity and limiting end use. This study focuses on the experimental evaluation of the impact and the fate of 14 inorganic contaminants (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Sb, Se, Sn, V and Zn) on Nannochloropsis salina growth. Microalgae were cultivated in photobioreactors illuminated at 984 &#181;mol m-2 sec-1 and maintained at pH 7 in a growth media polluted with inorganic contaminants at levels expected based on the composition found in commercial coal flue gas systems. Contaminants present in the biomass and the medium at the end of a 7 day growth period were analytically quantified through cold vapor atomic absorption spectrometry for Hg and through inductively coupled plasma mass spectrometry for As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Sb, Se, Sn, V and Zn. Results show N. salina is a sensitive strain to the multi-metal environment with a statistical decrease in biomass yieldwith the introduction of these contaminants. The techniques presented here are adequate for quantifying algal growth and determining the fate of inorganic contaminants.

Integration of microalgal cultivation with industrial flue gas will ultimately introduce heavy metals and other inorganic compounds into the growth media. This study presents a procedure used to determine the end fate and impact of heavy metals and inorganic contaminants on the growth of Nannochloropsis salina grown in photobioreactors.

중금속과 미세 조류의 생산성에 다른 무기 오염 물질의 정량화

Increasing demand for renewable fuels has researchers investigating the feasibility of alternative feedstocks, such as microalgae. Inherent advantages ...

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron (Oxy)Hydroxides, Trace Elements, and Bacteria

Fate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as redox potential (Eh) and pH, but also to microbial activities that can play a direct or indirect role on speciation and/or mobility. Indeed, some bacteria can directly oxidize As(III) to As(V) or reduce As(V) to As(III). Likewise, bacteria are strongly involved in Hg cycling, either through its methylation, forming the neurotoxin monomethyl mercury, or through its reduction to elemental Hg&#176;. The fates of both As and Hg are also strongly linked to soil or aquifer composition; indeed, As and Hg can bind to organic compounds or (oxy)hydroxides, which will influence their mobility. In turn, bacterial activities such as iron (oxy)hydroxide reduction or organic matter mineralization can indirectly influence As and Hg sequestration. The presence of sulfate/sulfide can also strongly impact these particular elements through the formation of complexes such as thio-arsenates with As or metacinnabar with Hg.
Consequently, many important questions have been raised on the fate and speciation of As and Hg in the environment and how to limit their toxicity. However, due to their reactivity towards aquifer components, it is difficult to clearly dissociate the biogeochemical processes that occur and their different impacts on the fate of these TE.
To do so, we developed an original, experimental, column setup that mimics an aquifer with As- or Hg-iron-oxide rich areas versus iron depleted areas, enabling a better understanding of TE biogeochemistry in anoxic conditions. The following protocol gives step by step instructions for the column set-up either for As or Hg, as well as an example with As under iron and sulfate reducing conditions.

Experimental Column Setup for Studying Anaerobic Biogeochemical Interactions Between Iron OxyHydroxides, Trace Elements, and Bacteria

Fate and speciation of arsenic and mercury in aquifers are closely related to physio-chemical conditions and microbial activity. Here, we present an original experimental column setup that mimics an aquifer and enables a better understanding of trace element biogeochemistry in anoxic conditions. Two examples are presented, combining geochemical and microbiological approaches.

(옥 시) 철 수 산화물, 성분, 및 박테리아 혐 기성 생물 지구 화학적 상호 작용을 공부에 대 한 실험 열 설정

Fate and speciation of trace elements (TEs), such as arsenic (As) and mercury (Hg), in aquifers are closely related to physio-chemical conditions, such as ...

Self-standing Electrochemical Set-up to Enrich Anode-respiring Bacteria On-site

Anaerobic respiration coupled with electron transport to insoluble minerals (referred to as extracellular electron transport [EET]) is thought to be critical for microbial energy production and persistence in many subsurface environments, especially those lacking soluble terminal electron acceptors. While EET-capable microbes have been successfully isolated from various environments, the diversity of bacteria capable of EET is still poorly understood, especially in difficult-to-sample, low energy or extreme environments, such as many subsurface ecosystems. Here, we describe an on-site electrochemical system to enrich EET-capable bacteria using an anode as a respiratory terminal electron acceptor. This anode is connected&#160;to a cathode capable of&#160;catalyzing abiotic oxygen reduction. Comparing this approach with electrocultivation methods that use a potentiostat for poising the electrode potential, the two-electrode system does not require an external power source. We present an example of our on-site enrichment utilized in an alkaline pond at the Cedars, a terrestrial serpentinization site in Northern California. Prior attempts to cultivate mineral reducing bacteria were unsuccessful, which is likely due to the low-biomass nature of this site and/or the low relative abundance of metal reducing microbes. Prior to implementing our two-electrode enrichment, we measured the vertical profile of dissolved oxygen concentration. This allowed us to place the carbon felt anode and platinum-electroplated carbon felt cathode at depths that would support anaerobic and aerobic processes, respectively. Following on-site incubation, we further enriched the anodic electrode in the laboratory and confirmed&#160;a distinct microbial community compared to the surface-attached or biofilm communities normally observed at the Cedars. This enrichment subsequently led to the isolation of the first electrogenic microbe from the Cedars. This method of on-site microbial enrichment has the potential to greatly enhance the isolation of EET-capable bacteria from low biomass or difficult to sample habitats.

On-site microbial enrichment or in situ cultivation techniques can facilitate the isolation of difficult-to-culture microbial taxa, especially from low-biomass or geochemically extreme environments. Here, we describe an electrochemical set-up without using an external power source to enrich microbial strains that are capable of extracellular electron transport (EET).

현장 양극 다 박테리아를 풍부 하 게 각자 서 있는 전기 설정

Anaerobic respiration coupled with electron transport to insoluble minerals (referred to as extracellular electron transport [EET]) is thought to be ...

Microalgae Cultivation and Biomass Quantification in a Bench-Scale Photobioreactor with Corrosive Flue Gases

Photobioreactors are illuminated cultivation systems for experiments on phototrophic microorganisms. These systems provide a sterile environment for microalgal cultivation with temperature, pH, and gas composition and flow rate control. At bench-scale, photobioreactors are advantageous to researchers studying microalgal properties, productivity, and growth optimization. At industrial scales, photobioreactors can maintain product purity and improve production efficiency. The video describes the preparation and use of a bench-scale photobioreactor for microalgal cultivation, including the safe use of corrosive gas inputs, and details relevant biomass measurements and biomass productivity calculations. Specifically, the video illustrates microalgal culture storage and preparation for inoculation, photobioreactor assembly and sterilization, biomass concentration measurements, and a logistic model for microalgal biomass productivity with rate calculations including maximum and overall biomass productivities. Additionally, since there is growing interest in experiments to cultivate microalgae using simulated or real waste gas emissions, the video will cover the photobioreactor equipment adaptations necessary to work with corrosive gases and discuss safe sampling in such scenarios.

Bench-scale, axenic cultivation facilitates microalgal characterization and productivity optimization before subsequent process scale-up. Photobioreactors provide the necessary control for reliable and reproducible microalgal experiments and can be adapted to safely cultivate microalgae with the corrosive gases (CO2, SO2, NO2) from municipal or industrial combustion emissions.

부식성 연도 가스가 있는 벤치 스케일 광생물반응기에서 미세조류 재배 및 바이오매스 정량화

Photobioreactors are illuminated cultivation systems for experiments on phototrophic microorganisms. These systems provide a sterile environment for ...

Simulation of Early Earth Hydrothermal Chimneys in a Thermal Gradient Environment

Deep sea hydrothermal vents are self-organizing precipitates generated from geochemical disequilibria and have been proposed as a possible setting for the emergence of life. The growth of hydrothermal chimneys in a thermal gradient environment within an early Earth vent system was successfully simulated by using different hydrothermal simulants, such as sodium sulfide, which were injected into an early Earth ocean simulant containing dissolved ferrous iron. Moreover, an apparatus was developed to sufficiently cool the ocean simulant to near 0 &#176;C in a condenser vessel immersed in a cold water bath while injecting a sulfide solution at hot to room temperatures, effectively creating an artificial chimney structure in a temperature gradient environment over a period of a few hours. Such experiments with different chemistries and variable temperature&#160;gradients resulted in a variety of morphologies in the chimney structure. The use of ocean and hydrothermal fluid simulants at room temperature resulted in vertical chimneys, whereas the combination of a hot hydrothermal fluid and cold ocean simulant inhibited the formation of robust chimney structures. The customizable 3D printed condenser created for this study acts as a jacketed reaction vessel that can be easily modified and used by different researchers. It will allow the careful control of injection rate and chemical composition of vent and ocean simulants, which should help accurately simulate prebiotic reactions in chimney systems with thermal gradients similar to those of natural systems.

The goal of this protocol is to form simulated hydrothermal chimneys via chemical garden injection experiments and introduce a thermal gradient across the inorganic precipitate membrane, using a 3D printable condenser that can be reproduced for educational purposes.

열그라이드 환경에서 초기 지구 수열 굴뚝 시뮬레이션

Deep sea hydrothermal vents are self-organizing precipitates generated from geochemical disequilibria and have been proposed as a possible setting for the ...

Using Flexible Gold-Titanium Reaction Cells to Simulate Pressure-Dependent Microbial Activity in the Context of Subsurface Biomining

Laboratory studies investigating subsurface microbial processes, such as metal leaching in deep ore deposits (biomining), share common and challenging obstacles, including the special environmental conditions that need to be replicated, e.g., high pressure and in some cases acidic solutions. The former requires an experimental setup suitable for pressurization up to 100 bar, while the latter demands a fluid container with high chemical resistance against corrosion and unwanted chemical reactions with the container wall. To meet these conditions for an application in the field of in situ biomining, a special flexible gold-titanium reaction cell inside a rocking high-pressure reactor was used in this study. The described system allowed simulation of in situ biomining through sulfur-driven microbial iron reduction in an anoxic, pressure-controlled, highly chemically inert experimental environment. The flexible gold-titanium reaction cell can accommodate up to 100 mL of sample solution, which can be sampled at any given time point while the system maintains the desired pressure. Experiments can be performed on timescales ranging from hours to months. Assembling the high-pressure reactor system is fairly time consuming. Nevertheless, when complex and challenging (microbiological) processes occurring in the earth&#39;s deep subsurface in chemically aggressive fluids have to be investigated in the laboratory, the advantages of this system outweigh the disadvantages. The results found that even at high pressure the microbial consortium is active, but at significantly lower metabolic rates.

This protocol describes microbial experiments under elevated pressures to study in situ biomining processes. The experimental approach employs a rocking high-pressure reactor equipped with a gold-titanium reaction cell containing a microbial culture in an acidic, iron-rich medium.

유연한 금-티타늄 반응 세포를 사용하여 지하 생물 광의 맥락에서 압력 의존미생물 활성 시뮬레이션

Laboratory studies investigating subsurface microbial processes, such as metal leaching in deep ore deposits (biomining), share common and challenging ...

공학

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic performance of microalgae-based biofuel production. Models that estimate microalgal growth under different conditions can help to optimize PBR design and operation. To be effective, the growth parameters used in these models must be accurately determined. Algal growth experiments are often constrained by the dynamic nature of the culture environment, and control systems are needed to accurately determine the kinetic parameters. The first step in setting up a controlled batch experiment is live data acquisition and monitoring. This protocol outlines a process for the assembly and operation of a bench-scale photosynthetic bioreactor that can be used to conduct microalgal growth experiments. This protocol describes how to size and assemble a flat-plate, bench-scale PBR from acrylic. It also details how to configure a PBR with continuous pH, light, and temperature monitoring using a data acquisition and control unit, analog sensors, and open-source data acquisition software.

This paper describes the assembly process and operation of a bench-scale photosynthetic bioreactor that can be used, in conjunction with other methods, to estimate pertinent kinetic growth parameters. This system continuously monitors the pH, light, and temperature using sensors, a data acquisition and control unit, and open-source data acquisition software.

키네 성장 시험을위한 온도, 빛 및 pH 모니터링을 갖춘 벤치 규모의 조류 광합성 생물 반응기의 건설 및 설치

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic ...

생체공학

Establishment of Microbial Eukaryotic Enrichment Cultures from a Chemically Stratified Antarctic Lake and Assessment of Carbon Fixation Potential

Lake Bonney is one of numerous permanently ice-covered lakes located in the McMurdo Dry Valleys, Antarctica. The perennial ice cover maintains a chemically stratified water column and unlike other inland bodies of water, largely prevents external input of carbon and nutrients from streams. Biota are exposed to numerous environmental stresses, including year-round severe nutrient deficiency, low temperatures, extreme shade, hypersalinity, and 24-hour darkness during the winter 1. These extreme environmental conditions limit the biota in Lake Bonney almost exclusively to microorganisms 2.
Single-celled microbial eukaryotes (called "protists") are important players in global biogeochemical cycling 3 and play important ecological roles in the cycling of carbon in the dry valley lakes, occupying both primary and tertiary roles in the aquatic food web. In the dry valley aquatic food web, protists that fix inorganic carbon (autotrophy) are the major producers of organic carbon for organotrophic organisms 4, 2. Phagotrophic or heterotrophic protists capable of ingesting bacteria and smaller protists act as the top predators in the food web 5. Last, an unknown proportion of the protist population is capable of combined mixotrophic metabolism 6, 7. Mixotrophy in protists involves the ability to combine photosynthetic capability with phagotrophic ingestion of prey microorganisms. This form of mixotrophy differs from mixotrophic metabolism in bacterial species, which generally involves uptake dissolved carbon molecules. There are currently very few protist isolates from permanently ice-capped polar lakes, and studies of protist diversity and ecology in this extreme environment have been limited 8, 4, 9, 10, 5. A better understanding of protist metabolic versatility in the simple dry valley lake food web will aid in the development of models for the role of protists in the global carbon cycle.
We employed an enrichment culture approach to isolate potentially phototrophic and mixotrophic protists from Lake Bonney. Sampling depths in the water column were chosen based on the location of primary production maxima and protist phylogenetic diversity 4, 11, as well as variability in major abiotic factors affecting protist trophic modes: shallow sampling depths are limited for major nutrients, while deeper sampling depths are limited by light availability. In addition, lake water samples were supplemented with multiple types of growth media to promote the growth of a variety of phototrophic organisms.
RubisCO catalyzes the rate limiting step in the Calvin Benson Bassham (CBB) cycle, the major pathway by which autotrophic organisms fix inorganic carbon and provide organic carbon for higher trophic levels in aquatic and terrestrial food webs 12. In this study, we applied a radioisotope assay modified for filtered samples 13 to monitor maximum carboxylase activity as a proxy for carbon fixation potential and metabolic versatility in the Lake Bonney enrichment cultures.

Microbial eukaryotes are both a source of photosynthetically-derived carbon and top predatory species in permanently ice-covered Antarctic lakes. This report describes an enrichment culture approach to isolate metabolically versatile microbial eukaryotes from the Antarctic lake, Lake Bonney, and assesses inorganic carbon fixation potential using a radioisotope assay for Ribulose-1,5-bisphophate carboxylase oxygenase (RubisCO) activity.

탄소 고정 가능성 화학 층상 남극 호수 및 평가에서 미생물 진핵 농축 문화의 확립

Lake Bonney is one of numerous permanently ice-covered lakes located in the McMurdo Dry Valleys, Antarctica. The perennial ice cover maintains a ...

생물학

Operation of Laboratory Photobioreactors with Online Growth Measurements and Customizable Light Regimes

The laboratory study of microalgae can be experimentally challenging. In addition to the cultivation requirements of non-photosynthetic microorganisms, phototrophs also require illumination. Routinely, researchers seek to provide custom light supplies, i.e., vary the light intensity and time over which it is delivered. Such flexibility is difficult with standard benchtop lights. Usually, cultivation studies also require growth comparisons between experimental treatments. Frequently, growth is assessed over an extended duration, e.g., multiple times a day over a week-long trial. Manual measurements can be time-consuming and lack data resolution. Therefore, photobioreactors (PBRs) with automatic growth monitoring and customizable light supply are useful for replicated experiments with multiple treatments. The current work presents the design, construction, and operation of laboratory PBRs. The materials are easily sourced and relatively inexpensive. The design could be duplicated with moderate skill. Each structure has a footprint of ~40 cm2 and hosts three 1 L glass bottles for triplicate replication. Bottles rest upon platforms containing magnetic stirrers and are arranged vertically within a 1 m high and 15 cm diameter polyvinyl chloride (PVC) pipe. The pipe interior is lined with light-emitting diodes (LEDs). These LEDs produce continuous light intensities from 0&#8211;2400 &#181;mol photons m-2 s-1 of photosynthetically active radiation (PAR). Users design a custom lighting program. The light intensity can be adjusted each second or held constant for longer durations. Oxygen produced from photosynthesis exits each bottle via a one-way volumetric gas sensor. Software is used to record gas sensor data. The amount of oxygen produced can be correlated to biomass growth. If biomass samples are required, a syringe can be used to extract culture. The method is suited for microalgae grown with bicarbonate as the carbon source. These PBRs are valuable to a laboratory that requires replicated experiments, light regime flexibility, and continuous high-resolution growth data.

This publication describes the design of laboratory photobioreactors (PBRs) with customizable light regimes. The growth of cyanobacteria or microalgae, using bicarbonate as their carbon source, is monitored continuously by measuring volumetric oxygen production. These PBRs facilitate rapid, replicated laboratory growth comparisons with little user intervention during experiments.

온라인 성장 측정 및 사용자 정의 가능한 조명 체계를 갖춘 실험실 광생물 반응기 운영

The laboratory study of microalgae can be experimentally challenging. In addition to the cultivation requirements of non-photosynthetic microorganisms, ...