Name: use of chironomidae (diptera) surface-floating pupal exuviae as a rapid bioassessment protocol for water bodies
Uploaded: 2015-07-24T14:00:00.000+00:00
Duration: 8 min 27 s
Description: Watch this Scientific Journal Video about Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies at JoVE.com

구성 및이 문서를 게시하기위한 자금 조달은 미네소타 대학에서 곤충학의학과 Chironomidae 연구 그룹 (LC Ferrington, 주니어, PI)에 다수의 보조금과 계약을 통해 제공되었습니다. 감사합니다 나단 로버츠에 수치로 사용 현장 사진을 공유 비디오에서이 원고와 관련.

필드 컬렉션 용품 1. 준비 <ol><li> 연구 설계에 기초하여 수집하고, 각 샘플에 대한 하나의 샘플 병 (예를 들어, 60 ml)에 취득한다 SFPE 샘플들의 수를 결정한다. </li><li> 각 샘플 항아리 두 개의 날짜와 지역 레이블을 준비합니다. 내부에 하나 배치하고 병의 외부로의 다른 부착. 샘플을 수집, 국가, 주, 카운티, 도시, 물, 몸, GPS 좌표, 날짜, 사람 (들)의 이름 : 각 날짜와 지역 레이블이 다음과 같은 정보가 포함되어 있는지 확인합니다. </li><li> 다른 특정 재료 및 장비 (특정 재료 / 장비의 표 참조)를 수집합니다. </li></ol> 2. 필드 컬렉션 <ol><li> 한 손으로 애벌레 트레이 및 다른 체를 잡으십시오. SFPE 축적 물에 애벌레 트레이를 찍어 (예를 들면, 거품 축적, 그루터기, 긴급 식물, 다시 파편, 소용돌이, 은행 가장자리를 따라) (그림 2A)는 와트를 허용어, exuviae, 이물질은 애벌레 트레이를 입력하고 체를 통해이 물질을 부어합니다. lotic 시스템의 샘플링, 샘플 범위의 하류 끝에서 시작하고 (그림 2B)를 상류 작업하는 경우. lentic 시스템의 샘플링하면, 바람 부는쪽으로 해안선에 시작합니다. <ol><li> 반복 각 미리 정의 된 샘플 범위 내에서 10 분 동안 2.1 단계 (또는 달리 특정 샘플링 정권에 대해 정의) (스트림에서 수집 한 샘플에 대한 일반적으로 100~200m,하지만 수중 모니터링 사이트의 전체 지역에 따라 다름); 적절한 SFPE 축적 영역 사이 이동합니다. </li></ol></li><li> 샘플 사이트에서 물이 가득 물총 병을 사용하여 체의 한 영역에서 이물질을 집중 조심스럽게 집게의 도움과 물총 병에서 에탄올의 흐름과 함께 SFPE 샘플에 사전 표지 샘플 항아리를 전송합니다. 에탄올 샘플 항아리를 채우십시오. </li><li> 반복 2.1 모든 시료 2.2 단계를. </li></ol> 3. 샘플 따기 참고 :이 프로토콜의 나머지 300 SFPE의 표본에 관한 다른 표본 크기에 맞게 수정해야 할 수도 있습니다. 연구 별 목표와 자원을 충족 SFPE 방법을 조정하는 보우 차드와 Ferrington의 9 서브 샘플링과 샘플링 주파수 지침을 참조하십시오. <ol><li> 각 SFPE 샘플에 대한 1-DRAM 용 유리 병을 할당; 각각의 유리 병 내부에 배치하고 에탄올로 가득 찬 유리 병을 ¾ 채우기 위해 날짜와 장소 레이블을 준비합니다. </li><li> 해당 샘플 항아리 뚜껑을 제거하고 부착 번데기 exuviae 확인합니다. 조심스럽게 에탄올로 가득 물총 병을 사용하여 페트리 접시에 뚜껑 내용을 씻어. 찾아 페트리 접시에 라벨 오프 내용을 씻어 부드럽게 집게를 사용하여 샘​​플 항아리의 내부에서 라벨을 제거하고. 따로 라벨을 설정합니다. </li><li> 더 SFPE 샘플 항아리에 남아 있도록 에탄올로 세척, 애벌레 트레이에 샘플 항아리의 내용을 전송합니다. 번데기 exuviae, RESI의 일부를 이동페트리 접시에 트레이에서 때문에, 에탄올. 샘플이 에탄올에 포함되어 있는지 확인합니다. </li><li> 스테레오 현미경 페트리 접시를 놓습니다. 체계적으로 번데기 exuviae의 페트리 접시의 내용을 검사합니다. 유리 병에 핀셋과 장소를 사용하여 요리의 모든 번데기 exuviae을 선택합니다. 부러 표본을 선택하지 마십시오, 건조, 이상 식별 문제를 방지하기 위해 압축 된 (즉, 두흉부와 복부의 절반 이상이 없습니다). 주 : 종 식별은 종종 어떤 경우에는, 속 레벨 식별 부분 표본으로 가능하다하더라도, 전체 시료가 존재하는 것을 필요로한다. <ol><li> 소용돌이 요리와 초기에 감지되지 않았을 수 있습니다 접시의 측면에 붙어있을 수있는 어떤 포함한 추가 번데기 exuviae뿐만 아니라, 어떤 작은 반투명 표본 검사. 두 개의 연속 스캔이 추가 번데기 exuviae을 공개하지 않을 때까지 반복합니다. </li></ol></li><li> 단계를 반복 3.33.4 전부 또는 300 번데기 exuviae가 포착 될 때까지. 300 번데기 exuviae이 포착되었을 때, 애벌레 트레이에 페트리 접시에서 잔류 물을 반환하고 에탄올로 페트리 접시를 씻어. 그런 다음, 빈 샘플 병에 애벌레 트레이에서 잔류 물을 전송 날짜와 지역 레이블을 추가하고, 항아리에 뚜껑을 넣어. 보관 또는 프로젝트 별 프로토콜에 따라 잔류 폐기하십시오. </li></ol> 정렬 4. 샘플 <ol><li> 모든 단지 표본을 커버하기에 충분한 에탄올로 가득 페트리 접시에 표시된 유리 병에서 번데기 exuviae 고른 따르십시오. </li><li> 스테레오 현미경, 분리 된 별개의 형태 학적 그룹 (즉, morphotaxa)으로 표본과는 별도로 표시 튜브가 에탄올로 일 3/4 완전 충전에 각 morphotaxon 놓습니다. <ol><li> chironomid의 morphotaxa 별도의 외부 형태 학적 특성을 활용합니다. 예를 들어, 두흉부에서, 존재, 크기, 형상의 차이를 이용하고,두부 결핵, 정면 사마귀, 정면의 세타, 흉부 뿔의 착색. 복부에서 morphotaxa 분리 (그림 4A)의 항문 로브뿐만 아니라, 복부 세그먼트의 등뼈, hookrows, 샤 그린, 세타, 그리고 박차를 사용합니다. Ferrington를 참조, 등. 5, Sæther 26 PINDER 추가 설명 및 형태 학적 특성의 수치에 대한 라이스 (27). </li><li> 표본은 건조하기 시작하면 추가로 에탄올을 사용합니다. </li></ol></li></ol> 설치 5. 슬라이드 <ol><li> 95 % 에탄올로 각각 morphotaxon 위해 멀티 웰 플레이트의 한 웰을 채운다. <ol><li> 여러 표현을 배치 (예를 들어, 전체의 25 %) 각 morphotaxon의 슬라이드 플레이트의 각 웰에 장착합니다. 표본이 충분히 탈수 적어도 10 분 동안 잘 앉을 수 있습니다. </li></ol></li><li> 해당 사이트, 수집 및 식별 많은 informati 레이블 슬라이드(그림 3)에. </li><li> 스테레오 현미경에 놓고 슬라이드. 참고 : 단계로 녹화 슬라이드 템플릿 일관된 배치하는 데 유용합니다. </li><li> 슬라이드에 Euparal 한 방울을 배치; 그것을 커버 슬립의 크기에 근사하도록 Euparal 확산. Euparal 작업시 적절한 환기를 사용합니다. 참고 : Euparal 작업시 적절한 환기를 사용합니다. </li><li> 집게를 사용 Euparal에 첫 번째 morphotaxon의 대표를 포함시킵니다. 참고 : 실험실 Euparal에서 그것을 포함하기 전에 잎사귀에 부드럽게 표본을 살짝 집게를 사용하여, 표본에서 초과 에탄올을 무효로합니다. </li><li> 뾰족한 집게 및 / 또는 해부 프로브 (그림 4A)를 사용하여 복부에서 두흉부를 분리합니다. <ol><li> ecdysial 봉합 (그림 4B)을 따라 두흉부를 분할하고 봉합 가장자리가 반대편 (그림 4C)에 있도록 두흉부를 엽니 다. </li><li> C의 방향ephalothorax이되도록 복부 측면 (그림 4C)를 직면하고있다. </li><li> 복부 지느러미 측면을 배치; 즉시 두흉부 (그림 4C) 아래에 배치합니다. </li></ol></li><li> 시편에 커버 슬립을 놓습니다. 하나의 에지가 천천히 아래 슬라이드를 감동과 함께, 각도로 커버 슬립을 잡고 공기 방울 형성​​을 줄이기 위해 커버 슬립을 놓습니다. 가볍게 커버 슬립에 눌러 시편을 평평하게합니다. </li><li> 반복 모든 탈수 표본 5.7을 통해 5.3 단계를 반복합니다. </li></ol> 6. 속 식별 <ol><li> 복합 현미경을 사용하여 슬라이드에 장착 된 표본의 속을 결정합니다. 등 Wiederholm 28 Ferrington에 키와 진단을 사용하여 속에 표본을 확인합니다. 5. 필요한 경우 등, Ferrington를 사용 가족 - 레벨 ID를 확인한다. (5). 참고 : Wiederholm 28 Ferrington 이후 다수의 일반적인 설명 및 개정이 계속있다,등. (5)는, 따라서, 이러한 키 및 진단은 불완전 일차 문헌으로 보충 될 필요가있다. </li></ol>

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M., Stur, E., Ekrem, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+and+Morphological+Methods+Reveal+Cryptic+Diversity+and+Three+New+Species+of+Nearctic+Micropsectra+(Diptera:+Chironomidae).">Molecular and Morphological Methods Reveal Cryptic Diversity and Three New Species of Nearctic Micropsectra (Diptera: Chironomidae).</a> Freshwater Science. 32, 892-921 (2013).</li><li>Ekrem, T., Willassen, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploring+Tanytarsini+Relationships+(Diptera:+Chironomidae)+using+Mitochondrial+COII+Gene+Sequences.">Exploring Tanytarsini Relationships (Diptera: Chironomidae) using Mitochondrial COII Gene Sequences.</a> Insect Systematics &amp; Evolution. 35, 263-276 (2004).</li><li>Ekrem, T., Willassen, E., Stur, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Comprehensive+DNA+Sequence+Library+is+Essential+for+Identification+with+DNA+Barcodes.">A Comprehensive DNA Sequence Library is Essential for Identification with DNA Barcodes.</a> Molecular Phylogenetics and Evolution. 43, 530-542 (2007).</li></ol>

저자는 그들이 더 경쟁 재정적 이익이 없다는 것을 선언합니다.

성공적인 SFPE 샘플 수집, 슬라이드 장착, 따기 분류 및 식별을위한 가장 중요한 단계는 다음과 같습니다 (1) 필드 컬렉션 (그림 2A) 동안 연구 지역 내에서 높은 SFPE 축적의 영역을 위치; (2)을 천천히 시료 채취시 모든 SFPE의 검출을위한 배양 접시의 내용을 주사; (3) (그림 4A)를 장착 슬라이드 동안 복부에서 두흉부를 해부하는 데 필요한 손재주를 개발; (4) 제대로 속에 식별 ?...

환경 상태를 평가하는 생물체를 사용하여 생물학적 모니터링 프로그램은 종종 수질을 평가 또는 생태계를 복원 프로그램의 성공을 모니터링하는 데 사용된다. 저서 성 대형 무척추 동물 군집을 사용하여 빠른 생물학적 평가 프로토콜 (RBP)은 1989 1 일부터 국가 수자원 기관 사이에 인기가있다. 예를 들면 딥-NET, Surber 샘플러, 헤스 샘플러 2와 RBPs에 대한 저서 성 대형 무척추 동물을 샘플링의 전통적인 방법, 시간 - 일 수있다 고가의 소비, 단지 특정 microhabitat 3에서 군집을 측정 할 수있다. 특정 수역에 대한 생물학적 정보를 생성하는 효율적인 대안 RBP는 Chironomidae 표면 부동 번데기 exuviae (SFPE) 3의 컬렉션을 포함한다. Chironomidae : 일반적으로 비 무는 작은 벌레로 알려진 (곤충강 파리목)는, 일반적으로 성인으로 부상하기 전에 수생 환경에서 발생 holometabolous 파리 있습니다<html> 60, 물 표면에. chironomid 제품군은 전 세계적으로 기술 된 약 5,000 종, 종 풍부하다; 그러나, 많은 20,000 종 4 존재하는 것으로 추정된다. Chironomids 때문에 높은 다양성과 변수 오염 허용 레벨 5의 많은 수중 생태계에 물과 서식지의 질을 문서화에 유용하다. 또한, 그들은 일반적으로 사회 5, 6에서 50 % 종 이상을 차지 자주 수생 시스템에서 가장 풍부하고 광범위한 저서 성 대형 무척추 동물이다. 지상파 성인의 출현에 따라, 번데기 exuviae (번데기 피부 캐스트) 물 표면에 떠 남아있다 (그림 1). 번데기 exuviae 바람 또는 물 전류의 작용을 통해 함께 은행이나 장애물 뒤에 축적 쉽고 빠르게 이전 24-48 시간 동안 7 등장 chironomid 종의 광범위한 샘플을 제공하기 위해 수집 될 수있다. 다른 사람이 5 매우 민감한 반면 ntent &quot;&gt; 상대 풍부하고 수집 된 SFPE의 분류 학적 조성물은 어떤 종은 허용 매우 오염되어 있음을 고려하여, 수질을 반영 SFPE 방법을 포함하여 기존의 애벌레 chironomid 샘플링 기술에 비해 많은 장점이 있습니다. (1) 최소 (2) 시료는 생물 수집에 초점을 맞출 것이 아니라 살아 있지 않은 피부, 그래서 지역 사회 역학의 궤도는 영향을받지 않는다; 속으로 (3) 식별 및있는 경우, 서식지 교란 샘플링 영역에서 발생 종종 종은, 상대적으로 쉽게 해당 키 및 설명 (3) 주어진 (4) 수집, 처리 및 샘플을 식별하는 기존의 샘플링 방법 3,8,9에 비해 효율적이고 경제적이다 (5) 축적 exuviae 유래 한 분류군을 나타냅니다 microhabitats (10)의 넓은 범위는 (6)에있어서 스트림 및 하천 하구, 락 비롯한 거의 모든 수중 환경에 적용 할 수있다ES, 연못, 바위 웅덩이, 습지, 그들은 모두 미성숙 단계를 완료하고 성공적으로 성인 (11)으로 부상 한 개인을 대표하기 때문에과 (7) SFPE 어쩌면 생태계 건강의 더 민감한 지표. SFPE 방법은 chironomid 지역 사회에 대한 정보를 수집하기위한 새로운 접근 방식이 아니다. SFPE의 사용은 먼저 1900 년대 초 Thienemann (12)에 의해 제안되었다. 연구의 다양한 (예 : 7,16-19) 분류 학적 조사 (예를 들어, 13 ~ 15), 생물 다양성 및 생태 학적 연구에 SFPE를 사용하고, 한 생물학적 평가 (예를 들어, 20 ~ 22). 또한, 일부 연구들은 표본 설계, 샘플 크기 및 종 또는 장군 (예, 8,9,23)의 다양한 검출 레벨을 달성하기 위해 필요한 샘플 이벤트 수의 다양한 측면을 다루었 다. 이러한 연구들은 종 또는 속의 비교적 높은 비율이 적당한 effor 검출 할 수 있음을 나타내t 또는 비용은 시료 처리와 연관된. 예를 들어, 앤더슨과 Ferrington 8은 100 카운트 표본을 기준으로 1/3 번째 적은 시간이 샘플 그물을 찍어 비교 SFPE 샘플을 선택하는 데 필요한 것을 결정했다. 또 다른 연구는 3-4 SFPE 샘플 분류 모든 DIP-NET 샘플 및 종 풍부가 3 증가로 SFPE 샘플 검출 종에서 딥-NET 샘플보다 더 효율적이라고 식별 할 수 있다고 결정했다. SFPE 샘플은 97.8 % (3) 효율적인 동안 예를 들어, 종 사이트에서 15 ~ 16 종의 값을 풍요 로움 평균 딥-NET 효율은 45.7 %이었다. 중요한 SFPE 방법은 유럽 연합 (EU) (24)에 규격화 된 생태 평가 북아메리카 25 (chironomid의 번데기의 exuviae 기법 (CPET)라고도 함), 그러나 본 방법은 상세하게 설명되지 않았다. SFPE 방법 중 하나는 응용 프로그램 등 Ferrington에 의해 설명되었다. <s&gt; 3을; 그러나, 연구의 주요 초점은 SFPE 방법의 효율성, 효과 및 경제를 평가하는 것이다. 이 연구의 목적은 샘플 수집, 처리 실험, 슬라이드 마운팅 및 속 식별 포함한 상세 SFPE 방법의 모든 단계를 설명하기위한 것이다. 대상은 자신의 연구로 기존의 수질 감시 활동을 확대에 관심있는 대학원생, 연구자 및 전문가를 포함한다.

<table><tbody><tr><td>Ethanol</td><td>Fisher Scientific</td><td>S25309B&amp;nbsp;</td><td>70-95%</td></tr><tr><td>Plastic wash bottles</td><td>Fisher Scientific</td><td>0340923B</td><td></td></tr><tr><td>Sample jar</td><td>Fisher Scientific</td><td>0333510B</td><td>Glass or plastic, 60-mL recommended</td></tr><tr><td>Testing sieve</td><td>Advantech</td><td>120SS12F</td><td>125-micron mesh size</td></tr><tr><td>Larval tray</td><td>BioQuip</td><td>5524</td><td>White</td></tr><tr><td>Stereo microscope</td><td></td><td></td><td></td></tr><tr><td>Glass shell vials</td><td>Fisher Scientific</td><td>0333926B</td><td>1-dram size</td></tr><tr><td>Plastic dropper</td><td>Thermo Scientific</td><td>1371110</td><td>30 to 35 drops/mL</td></tr><tr><td>Fine forceps</td><td>BioQuip</td><td>4524</td><td>#5</td></tr><tr><td>Petri dish</td><td>Carolina</td><td>741158</td><td>Glass or plastic</td></tr><tr><td>Multi-well plate</td><td>Thermo Scientific</td><td>144530</td><td>Glass or plastic</td></tr><tr><td>Glass microslides</td><td>Thermo Scientific</td><td>3010002</td><td>3 x 1 in.</td></tr><tr><td>Glass cover slips</td><td>Thermo Scientific</td><td>12-519-21G</td><td>Circular or square</td></tr><tr><td>Euparal mounting medium&amp;nbsp;</td><td>BioQuip</td><td>6372B</td><td></td></tr><tr><td>Pigma pen</td><td>BioQuip</td><td>1154F</td><td>Black</td></tr><tr><td>Probe</td><td>BioQuip</td><td>4751</td><td></td></tr><tr><td>Kimwipes</td><td>Kimberly-Clark Professional&amp;trade;</td><td>34120</td><td></td></tr></tbody></table>

use of chironomidae (diptera) surface-floating pupal exuviae as a rapid bioassessment protocol for water bodies

저서 대형 무척추 동물 군집을 사용하여 신속한 생물학적 평가 프로토콜이 성공적 수질에 인간의 영향을 평가하기 위해 사용되어왔다. 불행하게도, 그러한 딥 그물 같은 전통적인 저서 유생 샘플링 방법은, 시간 소모적이고 고가 일 수있다. 다른 프로토콜은 Chironomidae 표면 부동 번데기의 exuviae (SFPE)의 컬렉션을 포함한다. Chironomidae은 미성숙 단​​계 일반적으로 수생 서식지에서 발생 파리 (파리목)의 종 풍부한 제품군입니다. 성인 chironomids는, 물에서 나오는 물 표면에 떠있는, 자신의 번데기 스킨, 또는 exuviae를 떠나. Exuviae 종종 제방을 따라하거나이 chironomid 다양성과 풍요 로움을 평가하기 위해 수집 할 수있는 바람이나 물 전류의 작용에 의해 장애물 뒤에 축적된다. 어떤 종은 다른 사람보다 오염에 더 관대하기 때문에 Chironomids는 중요한 생물학적 지표로 사용할 수 있습니다. 따라서, 수집 된 SFPE의 상대 풍부 종 구성 반영수질의 변화. 여기서, 필드의 수집, 처리 실험, 슬라이드 마운팅 및 chironomid SFPE의 식별과 관련된 방법이 상세히 설명된다. SFPE 방법의 장점은 샘플링 영역에서 최소한의 방해, 효율적이고 경제적 인 시료 채취 및 실험실 처리, 식별의 용이성, 적용 거의 모든 수중 환경 및 생태계 스트레스를 잠재적으로 더 민감한 측정을 포함한다. 제한들이 성인 남성과 연관되지 않은 경우에 종 exuviae 번데기를 식별 유생 microhabitat 사용과 무능력을 판별 할 수없는 점 등이있다.

도 1은 chironomid 기간을 나타낸다 미성숙 단​​계 (계란, 애벌레, 번데기는) 일반적으로 발생하는, 또는 밀접하게 수생 환경과 관련된. 애벌레 삶의 단계가 완료되면, 유충은 튜브 같은 피난처를 구성하고 주변의 기판에 비단 분비물과 붙어서 번데기가 발생합니다. 현상 성인 성숙되면 번데기 자체를 해제하고 성인 번데기 exuviae으로부터 나올 수 물의 표면에 아무것도. exuviae 공기로 ?...

Watch this Scientific Journal Video about Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies at JoVE.com

Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

Rapid bioassessment protocols using benthic macroinvertebrates are often used to monitor and assess water quality. An efficient protocol involves collections of Chironomidae surface-floating pupal exuviae (SFPE). Here, techniques for field collection, laboratory processing, slide mounting, and identification of Chironomidae SFPE are described.

Chironomidae (파리목)의 사용 표면 부동 물 기관에 대한 신속한 생물학적 평가 프로토콜로 번데기 Exuviae을

Rapid bioassessment protocols using benthic macroinvertebrate assemblages have been successfully used to assess human impacts on water quality. ...

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Universidad de Cartagena UDEC

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JoVE Journal

Environment

Use of Chironomidae (Diptera) Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

11.6K 조회수.  University of Minnesota. Rapid bioassessment protocols using benthic macroinvertebrates are often used to monitor and assess water quality. An efficient protocol involves collections of Chironomidae surface-floating pupal exuviae (SFPE). Here, techniques for field collection, laboratory processing, slide mounting, and identification of Chironomidae SFPE are described.

비디오: Use of Chironomidae Diptera Surface-Floating Pupal Exuviae as a Rapid Bioassessment Protocol for Water Bodies

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP, 개선 INEX 방법에 대한<em&gt; 현장에서</em&gt; 미립자의 측정 및 용존 화합물은 액티브 서스펜션 피더에 의해 처리

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

연구

환경과학

Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may be treated with simple systems designed to promote sorption of contaminants to vegetation and soils and promote infiltration. Two example systems are described: a bioswale treatment system for urban stormwater treatment, and a vegetated drainage ditch for treating agriculture irrigation runoff. Both have similar attributes that reduce contaminant loading in runoff: vegetation that results in sorption of the contaminants to the soil and plant surfaces, and water infiltration. These systems may also include the integration of granulated activated carbon as a polishing step to remove residual contaminants. Implementation of these systems in agriculture and urban watersheds requires system monitoring to verify treatment efficacy. This includes chemical monitoring for specific contaminants responsible for toxicity. The current paper emphasizes monitoring of current use pesticides since these are responsible for surface water toxicity to aquatic invertebrates.

This article summarizes the design attributes and the effectiveness of treatment systems that treat urban stormwater and agriculture irrigation runoff to remove pesticides and other contaminants associated with aquatic toxicity.

농업과 도시 유출수의 지표수 독성과 관련된 오염 물질 제거를위한 식물 처리 시스템

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may ...

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical data that may be used in predictive modelling to forecast evolutionary responses of natural ecosystems to future environmental changes. However, excluding a few exceptional cases, long-term studies are scarce because of logistic difficulties associated with accessing temporal samples. Temporal dynamics are frequently studied in the laboratory or in controlled mesocosm experiments with exceptional studies that reconstruct the evolution of natural populations in the wild.
Here, a standard operating procedure (SOP) is provided to revive or resurrect dormant Daphnia magna, a widespread zooplankton keystone species in aquatic ecosystems, to dramatically advance the state-of-the-art longitudinal data collection in natural systems. The field of Resurrection Ecology was defined in 1999 by Kerfoot and co-workers, even though the first attempts at hatching diapausing zooplankton eggs date back to the late 1980s. Since Kerfoot&#39;s seminal paper, the methodology of resurrecting zooplankton species has been increasingly frequently applied, though propagated among laboratories only via direct knowledge transfer. Here, an SOP is described that provides a step-by-step protocol on the practice of resurrecting dormant Daphnia magna eggs.
Two key studies are provided in which the fitness response of resurrected Daphnia magna populations to warming is measured, capitalizing on the ability to study historical and modern populations in the same settings. Finally, the application of next generation sequencing technologies to revived or still dormant stages is discussed. These technologies provide unprecedented power in dissecting the processes and mechanisms of evolution if applied to populations that have experienced changes in selection pressure over time.

Resurrection of Dormant Daphnia magna: Protocol and Applications

Long-term studies are essential to understanding the process of evolution and the mechanisms of adaptation. Generally, these studies require commitments beyond the life-time of researchers. Here, a powerful method is described that dramatically advances state-of-the-art data collection to generate longitudinal data in natural systems.

부활의 휴면 벼룩 마그나: 프로토콜 및 응용 프로그램

Long-term studies enable the identification of eco-evolutionary processes that occur over extended time periods. In addition, they provide key empirical ...

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol for performing in situ juvenile exposure tests within oligotrophic river catchments over one-month and three-month periods. Two methods (in both modifications) are presented to evaluate the juvenile growth and survival rate. The methods and modifications differ in value for the locality bioindication and each has its benefits as well as limitations. The sandy cage method works with a large set of individuals, but only some of the individuals are measured and the results are evaluated in bulk. In the mesh cage method, the individuals are kept and measured separately, but a low individual number is evaluated. The open water exposure modification is relatively easy to apply; it shows the juvenile growth potential of sites and can also be effective for water toxicity testing. The within-bed exposure modification needs a high workload but is closer to the conditions of a natural juvenile environment and it is better for reporting the real suitability of localities. On the other hand, more replications are needed in this modification due to its high-hyporheic environment variability.

Bioindication Testing of Stream Environment Suitability for Young Freshwater Pearl Mussels Using In Situ Exposure Methods

In situ bioindications enable determination of the suitability of an environment for endangered mussel species. We describe two methods based on the juvenile exposure of freshwater pearl mussels in cages to oligotrophic river habitats. Both methods are implemented in variants for open water and hyporheic water environments.

Bioindication 영 민물 진주 홍합에 현장 노출 방법을 사용 하 여에 대 한 스트림 환경 적합성의 테스트

Knowledge of habitat suitability for freshwater mussels is an important step in the conservation of this endangered species group. We describe a protocol ...

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less attention has been paid on how environmental factors affect these insects when they pupate within the soil. The tea looper, Ectropis grisescens Warren (Lepidoptera: Geometridae), is a severe pest of tea plants and has caused huge economic losses in South China. The protocols described here aim to investigate, through multiple-choice bioassays, whether mature last-instar E. grisescens larvae can discriminate soil variables such as the substrate type and moisture content, and determine, through no-choice bioassays, the impact of the substrate type and moisture content on pupation behaviors and the emergence success of E. grisescens. The results would enhance the understanding of the pupation ecology of E. grisescens and may bring insights into soil-management tactics for suppressing E. grisescens populations. In addition, these bioassays can be modified to study the influences of various factors on the pupation behaviors and survivorship of soil-pupating pests.

Choice and No-Choice Bioassays to Study the Pupation Preference and Emergence Success of Ectropis grisescens

Here, we present a protocol to investigate the pupation preference of mature larvae of Ectropis grisescens in response to soil factors (e.g., substrate type and moisture content) using choice bioassays. We also present a protocol of no-choice bioassays to determine the factors that affect the pupation behaviors and survivorship of E. grisescens.

선택 및 아니오 선택 생물 검정 Pupation 환경과 Ectropis grisescens 의 출현 성공 공부

Many insects live above the ground as larvae and adults and as pupate below the ground. Compared to the above-ground stages of their life cycles, less ...

A Rapid and Efficient Method to Dissect Pupal Wings of Drosophila Suitable for Immunodetections or PCR Assays

Wing development in Drosophila melanogaster is an ideal model for studying morphogenesis at tissue level. These appendages develop from a group of cells named wing imaginal discs formed during embryonic development. In the larval stages the imaginal discs grow, increasing its number of cells and forming monolayered epithelial&#160;structures. Inside the pupal case, the imaginal discs bud out and fold into bilayers along a line that becomes the future margin of the wing. During this process, the longitudinal primodia veins originate vein cells on the prospective dorsal and ventral surfaces of the wing. During the pupal stage the stripes of vein cells of each surface communicate in order to generate tight tubes; at the same time, the cross-veins begin their formation.
With the help of appropriate molecular markers, it is possible to identify the major elements composing the wing during its development. For this reason, the ability to accurately detect transcripts or proteins in this structure is critical for studying their abundance and localization related to the development process of the wing.
The procedure described here focuses on manipulating pupal wings, providing detailed instructions on how to dissect the wing during the pupal stage. The dissection of pupal tissue is more difficult to perform than their counterparts in third instar larvae. This is why this approach was developed, to obtain rapid and efficient high quality samples. Details of how to immunostain and mount these wing samples, to allow the visualization of proteins or cell components, are provided in the protocol. With little expertise it is possible to collect 8-10 high quality pupal wings in a short amount of time.

A Rapid and Efficient Method to Dissect Pupal Wings of Drosophila Suitable for Immunodetections or PCR Assays

The ability to accurately detect transcripts or proteins in Drosophila tissues is critical for studying their abundance and localization related to the development process. Here is the description of a straightforward procedure to dissect pupal wings. These wings can be used as samples in several techniques (immunohistochemistry, PCR assay, etc.).

번데기 날개 초파리 Immunodetections 또는 PCR 분석 실험에 적합의 해 부를 신속 하 고 효율적인 방법

Wing development in Drosophila melanogaster is an ideal model for studying morphogenesis at tissue level. These appendages develop from a group of cells ...

발생학

Rapid Testing of Resistance of Timber to Biodegradation by Marine Wood-Boring Crustaceans

Wood-boring invertebrates rapidly destroy marine timbers and wooden coastal infrastructure, causing billions of dollars of damage around the globe every year. As treatments of wood with broad spectrum biocides, such as creosote and chromated copper arsenate (CCA), are now restricted in marine use by legislation, naturally durable timber species and novel preservation methods of wood are required. These methods undergo testing in order to meet regulatory standards, such as the European standard for testing wood preservatives against marine borers, EN 275. Initial investigation of durable timbers species or wood preservative treatments can be achieved quickly and inexpensively through laboratory testing, which offers many advantages over marine field trials that are typically costly, long-term endeavours. Many species of Limnoria (gribble) are marine wood-boring crustaceans. Limnoria are ideal for use in laboratory testing of biodegradation of wood by marine wood-borers, due to the practicality of rearing them in aquaria and the ease of measuring their feeding rates on wood. Herein, we outline a standardizable laboratory test for assessing wood biodegradation using gribble.

This protocol presents a method for assessing the feeding rate of the wood-boring crustacean, Limnoria, by measuring faecal pellet production. This method is designed for use in non-specialist labs and has potential for incorporation into standard testing protocols, to evaluate enhanced wood durability under marine conditions.

해양 목재-보링 갑각류에 의한 생분해에 대한 목재저항의 신속한 테스트

Wood-boring invertebrates rapidly destroy marine timbers and wooden coastal infrastructure, causing billions of dollars of damage around the globe every ...

Automatic Image Processing to Determine the Community Size Structure of Riverine Macroinvertebrates

Body size is an important functional trait that can be used as a bioindicator to assess the impacts of perturbations in natural communities. Community size structure responds to biotic and abiotic gradients, including anthropogenic perturbations across taxa and ecosystems. However, the manual measurement of small-bodied organisms such as benthic macroinvertebrates (e.g., &gt;500 &#181;m to a few centimeters long) is time-consuming. To expedite the estimation of community size structure, here, we developed a protocol to semi-automatically measure the individual body size of preserved river macroinvertebrates, which are one of the most commonly used bioindicators for assessing the ecological status of freshwater ecosystems. This protocol is adapted from an existing methodology developed to scan marine mesozooplankton with a scanning system designed for water samples. The protocol consists of three main steps: (1) scanning subsamples (fine and coarse sample size fractions) of river macroinvertebrates and processing the digitized images to individualize each detected object in each image; (2) creating, evaluating, and validating a learning set through artificial intelligence to semi-automatically separate the individual images of macroinvertebrates from detritus and artifacts in the scanned samples; and (3) depicting the size structure of the macroinvertebrate communities. In addition to the protocol, this work includes the calibration results and enumerates several challenges and recommendations to adapt the procedure to macroinvertebrate samples and to consider for further improvements. Overall, the results support the use of the presented scanning system for the automatic body size measurement of river macroinvertebrates and suggest that the depiction of their size spectrum is a valuable tool for the rapid bioassessment of freshwater ecosystems.

The article is based on the creation of an adapted protocol to scan, detect, sort, and identify digitized objects corresponding to benthic river macroinvertebrates using a semi-automatic imaging procedure. This procedure allows the acquisition of the individual size distributions and size metrics of a macroinvertebrate community in about 1 h.

강변 거대 무척추 동물의 군집 크기 구조를 결정하기위한 자동 이미지 처리

Body size is an important functional trait that can be used as a bioindicator to assess the impacts of perturbations in natural communities. Community ...

Hellgrammite as a Non-Conventional Biomonitor for Surface Water and Environmental Health Assessment

The larvae of dobsonflies from the genus Corydalus, commonly known as Hellgrammites, are characterized by their notable size, extensive range of occurrence, and extended period of immaturity, which can last up to one year. Hellgrammites are clearly known to exhibit sensitivity to pollution and habitat structure impacts. Given these unique features, using Corydalus texanus larvae is highly suitable as reliable biomonitoring agents to assess the ecological integrity of aquatic ecosystems. This protocol aims to provide the necessary tools for C. texanus assessment and demonstrate their efficacy through a case study. Research findings have practical implications, indicating that C. texanus larvae exhibit early-warning responses to mining pollution, bioaccumulating high amounts of heavy metals such as Zn, Fe, and Al. The presence or absence of C. texanus populations may serve as a helpful indicator for identifying potential issues related to ecosystem health. The unconventional approach has shown early warnings of pollution in mining-impacted sites, highlighting the need for timely action to protect the environment. Given their unique traits, the use of C. texanus larvae is highly suggested as a reliable non-conventional bioindicator.

This protocol describes the use of biomarkers for the early detection of deleterious impacts in aquatic ecosystems. The biomarkers are closely related to sentinel traits, and their changes aid in detecting early-warning damages.

The larvae of dobsonflies from the genus Corydalus, commonly known as Hellgrammites, are characterized by their notable size, extensive range of ...

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump&#39;s bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

This protocol describes the use of sump swabs and sludge analysis of zebrafish systems, which leads to increased detection compared to the sole use of sentinels to detect pathogens such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa. A system to monitor P. tomentosa eggs in quarantine is also proposed.

Chironomidae (파리목)의 사용 표면 부동 물 기관에 대한 신속한 생물학적 평가 프로토콜로 번데기 Exuviae을

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VacuSIP, 개선 INEX 방법에 대한