저자는 기꺼이 개발, 수정 및 저항과 너도밤 나무 규모의 곤충에 감염 될 수 있습니다 나무를 식별하기 위해 여기에 설명 된 프로토콜의 응용 프로그램을 지원 한 미국 산림청 산림 건강 보호 평가 모니터링 및 특별 기술 개발 프로그램의 자금 지원을 인정합니다. 우리는 또한 홀덴 수목원, 커틀 랜드에 감사하고, OH, 우리의 미국 너도밤 나무의 저항 검사 프로그램의 자신의 지속적인 협력과 지원을위한.

1 공장 물자 :. 성숙한 필드 나무, 화분에 심는 모종, 또는 화분 이식 <ol><li> 필드 테스트의 경우, 가능한 한 저항 테스트를위한 대규모 감염 또는 질병의 흔적을 보여 성숙한 건강한 미국 너도밤 나무를 선택합니다. 가시 나무 감수성 또한 제어 (도 1)로서 사용되는 식별 될 필요가있을 것이다. </li><li> 화분에 심은 모종 또는 이식을 테스트하기 위해, 수집하고 코흐 &amp; 캐리, 2004 년 캐리 등, 2013 년 한 바와 같이 이식 귀공자의 설명에 따라 beechnuts 발아. </li><li> 47g의 미량 영양소, 477g 완효성 비료 15N-3.9P-9.9K, 700g의 굵은 펄라이트 2.8 CU 당 75g 황산 알루미늄과 개정 토양 혼합 배양토에 모종이나 이식을 성장. 피트 가방. 성장시기에 나중에 필요할 경우, 200 ppm의 질소에 용해 17N-1.3P-14.1K 매주 식물을 비옥. </li><li> 성장시기 동안, 그늘 집에서 식물을 유지합니다. 공장 이전을 이동하는 가을에 외부 휴면 이동 허용온도 제어 저장 시설에 (~ 4 ° C) 4 월 중 11 월 ~. </li></ol> 너도밤 나무 규모 달걀 2. 컬렉션 <ol><li> BBD 득실 거리는 대에서 8 월 중순에 월 중순에서 일반적으로 풍부한 달걀의 존재를 확인하기 위해 손 돋보기 (쉽게 &quot;시키&quot;외관, 그림 1A에 의해 식별) 심하게 오염 된 나무를 검사합니다. </li><li> 부드럽게 플라스틱 밀봉 일갤런 수집 부대 (그림 1B)로, 성인 규모의 곤충, 계란과 다른 파편의 흰색 밀랍 덩어리를 브러시 페인트 브러시를 사용합니다. 적어도 12m 떨어져 세 가지 다른 나무의 최소에서 수집합니다. </li><li> 필요한 경우, 4 ℃에서 최대 두 주 동안 봉인 된 가방 컬렉션에 계란을 저장 테이프가 건조 계란을 방지하기 위해 가방의 내부에 진동 방지 발포 폴리에틸렌의 작은 조각 (2.5 cm의 사각형). </li><li> 성인 (0.60 mm)과 debri에서 규모 달걀 (0.15 × 0.25 mm)를 구분합니다의, 2 &#39;PVC 파이프의 짧은 조각 250 마이크론 나일론 메쉬의 정사각형 조각 (그림 2)을 지원하기 위해 커플 링을 사용하여 체를 구성. </li><li> 체에 성인, 왁스, 계란, 컬렉션 가방에서 파편의 혼합물을 비우고 가볍게 아래 유리 페트리 접시에 메쉬를 통과 계란을 장려하기 위해 작은 페인트 브러시를 사용합니다. 그들은 어려운 계란을 이동하고, 더 많은 정전기를 개최하기 때문에 플라스틱 페트리 접시는 피해야한다. 계란 전과 체질 후의도 3에 나타낸다. </li><li> 정제 한 알은 적어도 일주일 동안 39 ° C에서 배양 접시에 저장 될 수있다. 건조에서 계란을 방지하기 위해 뚜껑에 거품의 젖은 부분을 테이프와 파라 필름으로 밀봉. </li></ol> 3. 계란 생존 분석 실험 <ol><li> 달걀 생존력을 평가하기 위해, 60mm 유리 페트리 접시의 바닥의 주위 (도 4a 주위 바셀린의 얇은 링을 적용하는 10 CC 주사기를 사용 </stroNG&gt;). </li><li> , 링의 중심으로 약 100 계란 전송 페트리 접시에 뚜껑을 배치하고 파라 필름으로 밀봉. 계란 전에 분석을 시작으로 4 ° C에 보관 한 경우 밀봉 된 페트리 접시 2 주 또는 3 주 동안 실온에서 그대로 유지 할 수 있습니다. </li><li> 부화 약충은 석유 젤리에 갇혀 될 것입니다 쉽게 계산 될 수 있고, 빈 계란은 쉽게 자신의 색상과 광택 (그림 4B)에 의해 부화 계란에서 구별된다. 빈 계란 플러스 잔여 전체 달걀의 합 님프의 수를 나누어 생존을 계산한다. 좋은 가능성은 75 % ~ 90 % 부화 계란의 범위에 있어야합니다. </li></ol> 필드에 큰 성숙한 너도밤 나무 나무 4. 스케일 저항 검사 <ol><li> 정량 시험을 부드럽게 발포 폴리에틸렌의 predampened, 오픈 셀 (10) X 15 X 1.3 cm의 사각형의 중심을 가로 질러을 뿌려 해부 현미경을 사용하여 작은 주걱을 사용하여 500 계란을 계산합니다. 에거품을 저해 가능한 한 많은 물을 짜낸 후 젖은. 정성 시험, 500 계란 아웃 카운트 할 수 있고, 작은 유리 바이알 및 약 500 계란의 추가 배치를 측정 할 수있다 그려진 &quot;채우기&quot;라인에 배치. </li><li> 껍질에 직면 계란면 테스트 트리에 폼 패드를 놓습니다. 로프, 끈, 감기, 또는 플라스틱 코팅 하드웨어 와이어 장소에 패드를 누릅니다. 플라스틱 또는 금속 기반의 재료는 오히려 더 쉽게 야생 동물에 의해 소기 천연 섬유 소재보다 사용되어야한다. </li><li> 라인 맨과 아세테이트 기반의 실리콘 접착제와 수증기 투과성 방수 하우스 랩의 규격에 맞게 자르는 23 X 30cm 조각의 측면과는 거품 테스트 패드에 놓습니다. 방수 씰을 만들기 위해 나무에 집 랩의 가장자리를 누릅니다. 동안 접착제 세트 (그림 5)을 제자리에 고정하는, 나무와 집 랩의 주위에 나일론 꼬기 또는 플라스틱 코팅 하드웨어 와이어를 놓습니다. </li><li> 두 가지의 최소 배치바람직하게는 점토의 양측에 각각의 테스트 트리에 테스트 패드. 각 사이트에서 컨트롤로 (분명 자연 규모의 침략에) 적어도 2 감염 나무에 시험 패드를 배치합니다. 이전 제어 나무의 테스트 패드를 배치하는, 회사는 브러시를 곤두 사용하는 모든 자연적으로 발생하는 규모의 곤충이나 알을 제거합니다. 주 : 계란 거품 패드는 또한 침입 레벨이 낮은 영역에서 특히 유용 할 수있는 사육 치수 계란의 목적 나무에 배치 될 수있다. </li></ol> 화분에 심은 묘목 또는 이식의 5. 스케일 저항 검사 <ol><li> 테스트를 위해, 그들에 배치 된 적어도 2 개의 별도의 테스트 패드를 가질 정도로 키가 1cm (5cm 토양 라인 위에 캘리퍼스)의 최소 직경 화분에 심은 나무를 선택합니다. 필요한 경우 테스트 패드를위한 공간을 만들기 위해 작은 쪽 가지 치기. </li><li> 150 계란을 계산하고 규격에 맞게 자르는 통해 그들을 흩어 해부 현미경을 사용하여 2.5 X 7.6 X 1.3 cm 오픈 셀 폴리에틸렌 파이프를 적신렌 거품 패드. </li><li> 플라스틱 코팅 된 와이어를 사용하여, 나무 껍질에 계란 측과 모종에 폼 패드를 붙입니다. 다만 거품 위의 모종의 주위에 집 랩의 작은 사각형을 감싸고 (그림 6A 참조) 아세테이트 기반의 실리콘으로 밀봉. </li><li> 컨트롤로 취약 모종의 가족이나 이식 알려진 포함한다. </li></ol> 6. 데이터 수집 참고 : 약 52-57주 테스트 나무에 폼 패드와​​ 계란을 배치 한 후, 데이터를 수집 할 수 있습니다. 그것은이가 성인이 재현 할 수있는 능력을 판별 할 수 있도록 알을 낳기 시작했다 때까지 수행되지 않습니다하는 것이 중요합니다. 일부 저항 나무에 성인 적은 수의 확립 볼 드문되지 않습니다하지만 재생이 발생하지와. <ol><li> 조심스럽게 폼 패드를 제거하고 손으로 렌즈 나 돋보기 (10X)를 사용하여 껍질에 설립 된 성인 규모 곤충의 수를 계산합니다. </li><li> 폼 패드가 제거 될 때, U 아니다거품 나무의 탈옥하는 일부 성인과 대부분의 계란 클러스터 ncommon (그림 7 참조). 해부 현미경을 사용하여 계란 클러스터와 폼 패드에 부착 남아있다 성인을 계산합니다. </li></ol>

<ol>
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저자는 더 경쟁 재정적 이익이 없다는 것을 선언합니다.

이 분석의 성공을 위해 필요한 중요한 단계는 계란의 생존 능력 테스트 및 테스트를 각 사이트에서 민감한 컨트롤의 사용, 화분에 심는 재료를 수행 있습니다. 이는 트리 또는 모종 당 하나 이상의 테스트 패드를 사용하는 것도 중요하다. 우리는 필드와 화분에 심은 식물에 더 많은 제어 테스트에서 모두 오류의 잦은 소스 패드 실패 할 수 있다는 것을 발견했다. 예를 들어, 필드에, 심지어 물이 줄...

너도밤 나무 껍질 질병 (BBD)은 1890 년대 후반 노바 스코샤의 캐나다 지방의 침략 너도밤 규모의 곤충, 크립토 fagisuga의 도입 이후 북미에서 미국 밤나무에 해로운 영향을 미쳤습니다. 너도밤 나무 규모 곤충 곰팡이 (Neonectria의 ditissima 또는 Neonectria의 faginata)의 Neonectria 종 중 하나에 감염 입구의 통로를 제공하는 작은 균열을 만드는 나무 껍질로 그 먹이 탐침을 삽입 할 때이 곤충 질병 단지는 시작됩니다. 곰팡이 균사의 성장에 따라, 조직의 큰 영역은 결국 완전히 나무 가드 링, 죽을 수있다. 질병의 피해는 강풍 4 스냅에이 발생하기 쉬운 만들기, 나무를 약화. 질병의 첫 전파 사망률 수준은 50 % 5만큼 높은 것으로보고되었다. cankers는 나무 제품으로 나무의 가치를 감소시키는 형태로 남아있는 나무는 종종 심각하게 변형된다.0; 이러한 나무는, 설립 스탠드 (6)의 경제적, 생태 학적 가치를 감소에서 다른 더 바람직한 종을 방지 &quot;너도밤 나무 덤불&quot;의 형성에 이르게 루트 발아에 대한 성향이있다. 너도밤 나무 껍질 질병은 미국 너도밤 나무의 멸종으로 이어질 가능성이 아니지만, 그것은 야생 동물 7,8 음식과 서식지의 감소로 이어지는 구성과 건강을 서 변경합니다. 몇 년 동안 BBD의 영향을 스탠드에서, 질병의 증상 무료 남아 나무가보고되었습니다. 인공 접종 시험은이 나무가 규모의 곤충 2에 저항하는 것으로 나타났습니다. 규모 침입하지 않고, Neonectria 감염에 대한 약간의 기회가 곰팡이의 영향을 최소화 할 수있다. 이전 규모 감염의 부재에 Neonectria 감염으로 인해 미국의 너도밤 나무에서 대규모 사망은 결코 없다보고, 그래서 한 BBD에 대한 저항의 너도밤 나무 규모 곤충 결과에 대한 저항. BBD의 관리에 대한 최근의 연구는 식별, 전파, 번식, 그리고 너도밤 나무 규모의 곤충에 대한 저항과 미국의 너도밤 나무의 보존에 초점을 맞추고있다. 유전자 연구는 규모의 곤충에 대한 저항은 유전과 저항 나무의 신중한 선택과 번식이 하나의 세대 9 크게 향상 될 수있는 것으로 나타났습니다. 이 발견은 복원 재배 (10, 11)에 유 전적으로 다양한 BBD 저항 씨의 소스를 제공하기 위해 저항하는 미국 밤나무의 지역 씨 과수원을 설정하는 미국의 주 및 국가 산림 관리자의 노력을 자극했다. 연구는 또한 감염 나무와 저항 나무의 보존을 제거하여 스탠드 유전학의 조림 조작 스탠드 개선 9,12 발생할 수 있음을 지적했다. &lt;/ P&gt; 트리 개선 활동을 통해 또는 조림 처방전을 수행을 통해 BBD의 관리를위한 선택 너도밤 나무 스케일 방지 및 감염 나무를 구별 할 수있는 능력이 필요합니다. 여기에 제시된 방법은 먼저 인위적으로 너도밤 나무 규모의 계란 1 모종을 접종 데이브 휴스턴, 도입 방법에서 적응되었습니다. 이 방법은 저항 또는 유전자 연구에 저항하고 취약 화분 모종 또는 이식 ramets 구별하는 관련 양적 형질 유전자좌 (QTL)를 식별 할 수있는 검사 도구로 사용할 수 있습니다. 대안으로, 시드 필드 과수원 개발 또는 보존에 대해 저항하는 트리를 식별하는 필드 성숙한 나무의 스크리닝에 사용될 수있다. 감염되기 쉬운 나무 식별과 질병에 미치는 영향을 최소화하기 위해 제거 할 수 있습니다.

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			<td style="width:627px;">Mesh opening size: 250 microns square, % of open area: 34, thread diameter: 180 micron, width: 24&#8221;, Length: 12&#8221;, Package quantity: 1</td>
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a technique to screen american beech for resistance to the beech scale insect (cryptococcus fagisuga lind.)

크게 약화 및 변형 생존자 나무 뒤에 남겨두고 너도밤 나무 껍질 질병 초기 ​​사망률이 높은 수준에서 (BBD) 결과. 질병은 곰팡이의 Neonectria 종 중 하나에 의해 감염에 대한 진입 점을 만들어 침략 너도밤 규모의 곤충, 크립토 fagisuga의 활동을 공급함으로써 시작된다. 규모 침입하지 않고, 곰팡이 감염에 대한 약간의 기회가있다. 인위적으로 크게 BBD 영향을 스탠드에 건강한 나무를 만연하는 규모의 계란을 사용하면이 나무는 질병의 복잡한 1의 규모 곤충 부분에 저항 이었다는 것을 보여 주었다. 여기에서 우리는 우리가 현장에서 작은 화분에 심은 모종 이식의 스케일 방지 나무에 대한 화면으로 사용할 수있는 휴스턴에 의한 인공 감염의 기술을 기반으로 개발 한 프로토콜을 제시한다. 스케일 방지 나무의 식별은 나무를 통해 BBD의 관리의 중요한 구성 요소입니다개선 프로그램 및 조림 조작.

그림 6은 감수성의 다른 정도를 나타내는 저항 모종 (C)와 두 개의 취약 모종 (D, E)를 보여줍니다. 인공 감염 테스트를 설정 한 후 감염 성숙한 나무가 57주 모습의 예는 그림 (b)에 표시됩니다. 발포체가 얻어 트리로부터 박리되면 개각충 드물지가 아니며,도 7에 도시 된 바와 같이 남아 그들의 달걀 클러스터는 트리 폼 패드 모두로부터...

Watch this Scientific Journal Video about A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind. at JoVE.com

A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

너도밤 나무 껍질 질병은 껍질에 곰팡이 진입 점을 만들 너도밤 규모 곤충의 활동을 공급함으로써 시작된다. 스케일 곤충에 강한 나무는 질병에 저항하는. 여기에서 우리는 우리가 너도밤 나무 규모의 저항에 대한 개별 너도밤 나무를 화면에 개발 한 프로토콜을 제시한다.

비치 스케일 곤충에 대한 저항에 대한 미국 밤나무를 (화면에 기술<em&gt; 크립토 fagisuga</em&gt; 린드.)

Beech bark disease (BBD) results in high levels of initial mortality, leaving behind survivor trees that are greatly weakened and deformed.&#160;The ...

a-technique-to-screen-american-beech-for-resistance-to-beech-scale

Research

JoVE Journal

Environment

A Technique to Screen American Beech for Resistance to the Beech Scale Insect (Cryptococcus fagisuga Lind.)

9.5K 조회수.  US Forest Service.  너도밤 나무 껍질 질병은 껍질에 곰팡이 진입 점을 만들 너도밤 규모 곤충의 활동을 공급함으로써 시작된다. 스케일 곤충에 강한 나무는 질병에 저항하는. 여기에서 우리는 우리가 너도밤 나무 규모의 저항에 대한 개별 너도밤 나무를 화면에 개발 한 프로토콜을 제시한다. 

비디오: A Technique to Screen American Beech for Resistance to the Beech Scale Insect Cryptococcus fagisuga Lind.

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American lobsters. Recent work in exploited crustacean populations (crabs and lobsters) suggests that there are circumstances where mature females are unable to achieve their full reproductive potential due to sperm limitation. To examine this possibility in different regions of the American lobster fishery, a reliable and noninvasive method was developed for sampling large numbers of female lobsters at sea. This method involves inserting a blunt-tipped needle into the female&#39;s seminal receptacle to determine the presence or absence of a sperm plug and to withdraw a sample that can be examined for the presence of sperm. A series of control studies were conducted at the dock and in the laboratory to test the reliability of this technique. These efforts entailed sampling 294 female lobsters to confirm that the presence of a sperm plug was a reliable indicator of sperm within the receptacle and thus, mating. This paper details the methodology and the results obtained from a subset of the total females sampled. Of the 230 female lobsters sampled from George&#39;s Bank and Cape Ann, MA (size range = 71-145 mm in carapace length), 90.3% were positive for sperm. Potential explanations for the absence of sperm in some females include: immaturity (lack of physiological maturity), breakdown of the sperm plug after being used to fertilize a clutch of eggs, and lack of mating activity. The surveys indicate that this technique for examining the mating success of female lobsters is a reliable proxy that can be used in the field to document reproductive activity in natural populations.

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters Homarus americanus

Fishery-induced changes to exploited crustacean fisheries, such as the American lobster fishery, could potentially influence their reproductive dynamics, leading to a reduction in mating success. This study&#39;s goal was to develop a noninvasive method for ascertaining the mating success of female lobsters that may be physiologically or functionally mature.

에 대한 비 침습적 방법<em&gt; 현장에서</em&gt; 여성 미국의 바닷가에 연결 성공 (의 결정<em&gt; Homarus의 americanus</em&gt;)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American ...

연구

환경과학

Video Tracking Protocol to Screen Deterrent Chemistries for Honey Bees

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey bee colony numbers have been declining in the United States and many European countries since 1947. A number of factors play a role in this decline, including the unintentional exposure of honey bees to pesticides. The development of new methods and regulations are warranted to reduce pesticide exposures to these pollinators. One approach is the use of repellent chemistries that deter honey bees from a recently pesticide-treated crop. Here, we describe a protocol to discern the deterrence of honey bees exposed to select repellent chemistries. Honey bee foragers are collected and starved overnight in an incubator 15 h prior to testing. Individual honey bees are placed into Petri dishes that have either a sugar-agarose cube (control treatment) or sugar-agarose-compound cube (repellent treatment) placed into the middle of the dish. The Petri dish serves as the arena that is placed under a camera in a light box to record the honey bee locomotor activities using video tracking software. A total of 8 control and 8 repellent treatments were analyzed for a 10 min period with each treatment was duplicated with new honey bees. Here, we demonstrate that honey bees are deterred from the sugar-agarose cubes with a compound treatment whereas honey bees are attracted to the sugar-agarose cubes without an added compound.

The loss of honey bee colonies presents a challenge to crop pollination services. Current pollinator protection practices warrant an alternative approach to minimize the contact of honey bees to harmful pesticides using repellent chemistries. Here, we provide detailed methods for a visual tracking protocol to screen deterrents for bees.

꿀벌을위한 억제 성 화학 물질을 차단하는 비디오 추적 프로토콜

The European honey bee, Apis mellifera L., is an economically and agriculturally important pollinator that generates billions of dollars annually. Honey ...

Lab-Scale Model to Evaluate Odor and Gas Concentrations Emitted by Deep Bedded Pack Manure

A lab-scaled simulated bedded pack model was developed to study air quality and nutrient composition of deep-bedded packs used in cattle mono-slope facilities. This protocol has been used to effectively evaluate many different bedding materials, environmental variables (temperature, humidity), and potential mitigation treatments that can improve air quality in commercial deep-bedded mono-slope facilities. The model is dynamic and allows researchers to easily collect many chemical and physical measurements from the bedded pack. Weekly measurements, collected over the course of six to seven weeks, allows sufficient time to see changes in air quality measurements over time as the bedded pack matures. The data collected from the simulated bedded packs is within the range of concentrations previously measured in commercial deep-bedded mono-slope facilities. Past studies have demonstrated that 8 - 10 experimental units per treatment are sufficient to detect statistical differences among the simulated bedded packs. The bedded packs are easy to maintain, requiring less than 10 minutes of labor per bedded packs per week to add urine, feces, and bedding. Sample collection using the gas sampling system requires 20 - 30 minutes per bedded pack, depending on the measurements that are being collected. The use of lab-scaled bedded packs allows the researcher to control variables such as temperature, humidity, and bedding source that are difficult or impossible to control in a research or commercial facility. While not a perfect simulation of &#34;real-world&#34; conditions, the simulated bedded packs serve as a good model for researchers to use to examine treatment differences among bedded packs. Several lab-scale studies can be conducted to eliminate possible treatments before trying them in a research or commercial-sized facility.

A protocol has been developed to measure gases, odors, and nutrient composition in lab-scaled bedded manure packs, which can be used to study ways to improve air quality in commercial cattle facilities using deep-bedded manure packs.

냄새 및 가스 농도 깊은 방출 실험실 스케일 모델 침대 팩 비료

A lab-scaled simulated bedded pack model was developed to study air quality and nutrient composition of deep-bedded packs used in cattle mono-slope ...

Double-stranded RNA Oral Delivery Methods to Induce RNA Interference in Phloem and Plant-sap-feeding Hemipteran Insects

Phloem and plant sap feeding insects invade the integrity of crops and fruits to retrieve nutrients, in the process damaging food crops. Hemipteran insects account for a number of economically substantial pests of plants that cause damage to crops by feeding on phloem sap. The brown marmorated stink bug (BMSB), Halyomorpha halys (Heteroptera: Pentatomidae) and the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Liviidae) are hemipteran insect pests introduced in North America, where they are an invasive agricultural pest of high-value specialty, row, and staple crops and citrus fruits, as well as a nuisance pest when they aggregate indoors. Insecticide resistance in many species has led to the development of alternate methods of pest management strategies. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) is a gene silencing mechanism for functional genomic studies that has potential applications as a tool for the management of insect pests. Exogenously synthesized dsRNA or small interfering RNA (siRNA) can trigger highly efficient gene silencing through the degradation of endogenous RNA, which is homologous to that presented. Effective and environmental use of RNAi as molecular biopesticides for biocontrol of hemipteran insects requires the in vivo delivery of dsRNAs through feeding. Here we demonstrate methods for delivery of dsRNA to insects: loading of dsRNA into green beans by immersion, and absorbing of gene-specific dsRNA with oral delivery through ingestion. We have also outlined non-transgenic plant delivery approaches using foliar sprays, root drench, trunk injections as well as clay granules, all of which may be essential for sustained release of dsRNA. Efficient delivery by orally ingested dsRNA was confirmed as an effective dosage to induce a significant decrease in expression of targeted genes, such as juvenile hormone acid O-methyltransferase (JHAMT) and vitellogenin (Vg). These innovative methods represent strategies for delivery of dsRNA to use in crop protection and overcome environmental challenges for pest management.

This article demonstrates novel techniques developed for oral delivery of double-stranded RNA (dsRNA) through the vascular tissues of plants for RNA interference (RNAi) in phloem sap feeding insects.

체 관 부 식물 수액 공급 Hemipteran 곤충에 RNA 간섭을 유도 하 이중 가닥 RNA 구두 전달 방법

Phloem and plant sap feeding insects invade the integrity of crops and fruits to retrieve nutrients, in the process damaging food crops. Hemipteran ...

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly relevant as a model for the studies of osmoregulatory mechanisms because of its extreme tolerance to low salinity. It is also widely used as a model of settling biology, in particular in relation to antifouling research. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae for studies. A protocol for the all-year-round culturing of B. improvisus has been developed and a detailed description of all steps in the production line is outlined (i.e., the establishment of adult cultures on panels, the collection and rearing of barnacle larvae, and the administration of feed for adults and larvae). The description also provides guidance on troubleshooting and discusses critical parameters (e.g., the removal of contamination, the production of high-quality feed, the manpower needed, and the importance of high-quality seawater). Each batch from the culturing system maximally yields roughly 12,000 nauplii and can deliver four batches in a week, so up to almost 50,000 larvae per week can be produced. The method used to culture B. improvisus is, probably, to a large extent also applicable to other marine invertebrates with free-swimminglarvae. Protocols are presented for the dissection of various tissues from adults as well as the production of high-quality RNA for studies on gene expression. It is also described how cultured adults and reared cyprids can be utilized in a wide array of experimental designs for examining gene expression in relation to external factors. The use of cultured barnacles in gene expression is illustrated with studies of possible osmoregulatory roles of Na+/K+ ATPase and aquaporins.

The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression

The barnacle Balanus (Amphibalanus) improvisus is a model for studying osmoregulation and antifouling. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae. Here, a protocol for the all-year-round culturing of B. improvisus is described, including the production of larvae. The use of cultured barnacles in gene expression studies is illustrated.

해양 모델-따개 비 Balanus improvisus 경작 및 유전자 발현

Barnacles are marine crustaceans with a sessile adult and free-swimming, planktonic larvae. The barnacle Balanus (Amphibalanus) improvisus is particularly ...

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and toxicological studies. Using automated digital tracking platforms, recent efforts in neuropharmacology are leveraging larval fish locomotor behaviors to identify potential therapeutic targets for novel small molecules. Similar to these efforts, research in the environmental sciences and comparative pharmacology and toxicology is examining various behaviors of fish models as diagnostic tools in tiered evaluation of contaminants and real-time monitoring of surface waters for contaminant threats. Whereas the zebrafish is a popular larval fish model in the biomedical sciences, the fathead minnow is a common larval fish model in ecotoxicology. Unfortunately, fathead minnow larvae have received considerably less attention in behavioral studies. Here, we develop and demonstrate a behavioral profile protocol using caffeine as a model neurostimulant. Though photomotor responses of fathead minnows were occasionally affected by caffeine, zebrafish&#160;were markedly more sensitive for photomotor and locomotor endpoints, which responded at environmentally relevant levels. Future studies are needed to understand comparative behavioral sensitivity differences among fish with age and time of day, and to determine whether similar behavioral effects would occur in nature and be indicative of adverse outcomes at the individual or population levels of biological organization.

Here, we present a protocol to examine larval zebrafish and fathead minnow locomotor activities and photomotor responses (PMR) using an automated tracking software. When incorporated in common toxicology bioassays, analyses of these behaviors provide a diagnostic tool to examine chemical bioactivity. This protocol is described using caffeine, a model neurostimulant.

애벌레에서 행동 응답 프로필을 조사 하기 위한 실험 프로토콜: 신경 흥분 제 카페인에 응용

Fish models and behaviors are increasingly used in the biomedical sciences; however, fish have long been the subject of ecological, physiological and ...

A Precise and Autonomous System for the Detection of Insect Emergence Patterns

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of emerging insects they can detect. In order to obtain precise measurement of insect emergence, it is necessary for systems to be semi-automated and able to measure large numbers of emerging insects. We addressed these issues by designing and building a system that is automated and can measure emergence of up to 1200 insects. We modified the existing &#34;falling-ball&#34; system using Arduino microcontrollers to automate data collection and expand the sample size through multiple data channels. Multiple data channels enable the user to not only increase their sample size, but also allows for multiple treatments to be run simultaneously in a single experiment. Furthermore, we created an R script to automatically visualize the data as a bubble plot, while also calculating the median day and time of emergence. The current system was designed using 3D printing so the user can modify the system to be adjusted for different species of insects. The goal of this protocol is to investigate important questions in chronobiology and stress physiology, using this precise and automated system to measure insect emergence patterns.

Measurement of insect emergence patterns requires precision. Existing systems are only semi-automated and sample size is limited. We addressed these issues by designing a system using microcontrollers to precisely measure the time of emergence of large numbers of emerging insects.

곤충 출현 패턴의 검출을 위한 정확 하 고 자치 시스템

Existing systems to measure insect emergence patterns have limitations; they are only partially automated and are limited in the maximum number of ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, we described a protocol for multiple analyses of the biological compositions in environmental PM. Five experiments are presented: (1) PM number monitoring by using a laser particle counter; (2) PM collection by using a cyclonic aerosol sampler; (3) PM collection by using a high-volume air sampler with filters; (4) culturable microbes collection by the Andersen six-stage sampler; and (5) detection of biological composition of environmental PM by bacterial 16SrDNA and fungal ITS region sequencing. We selected hazy days and a livestock farm as two typical examples of the application in this protocol. In this study, these two sampling methods, cyclonic aerosol sampler and filter sampler, showed different sampling efficiency. The cyclonic aerosol sampler performed much better in terms of collecting bacteria, while these two methods showed the same efficiency in collecting fungi. Filter samplers can work under low temperature conditions while cyclonic aerosol samplers have a sampling limitation for temperature. A solid impacting sampler, such as an Andersen six-stage sampler, can be used to sample bioaerosols directly into the culture medium, which increases the survival rate of culturable microorganisms. However, this method mainly relies on culture while more than 99% of microbes cannot be cultured. DNA extracted from the culturable bacteria collected by the Andersen six-stage sampler and samples collected by cyclonic aerosol sampler and filter sampler were detected by bacterial 16S rDNA and fungal ITS region sequencing.All the methods above may have wide application in many fields of study, such as environmental monitoring and airborne pathogen detection. From these results, we can conclude that these methods can be used under different conditions and may help other researchers further explore the health impacts of environmental bioaerosols.

Understanding the biological composition of environmental particulate matter is important for the study of its significant impacts on human health and disease spread. Here, we used three types of bioaerosol sampling methods and a biological analysis of airborne microbes to better explore airborne microbial communities under different environmental conditions.

구성 및 다른 환경 조건에서 Bioaerosols의 유통 분석

Variable microorganisms in particulate matter (PM) under different environmental conditions may have significant impacts on human health. In this study, ...

Detached Leaf Assays to Simplify Gene Expression Studies in Potato During Infestation by Chewing Insect Manduca sexta

The multitrophic nature of gene expression studies of insect herbivory demands large numbers of biological replicates, creating the need for simpler, more streamlined herbivory protocols. Perturbations of chewing insects are usually studied in whole plant systems. While this whole organism strategy is popular, it is not necessary if similar observations can be replicated in a single detached leaf. The assumption is that basic elements required for signal transduction are present within the leaf itself. In the case of early events in signal transduction, cells need only to receive the signal from the perturbation and transmit that signal to neighboring cells which are assayed for gene expression.
The proposed method simply changes the timing of the detachment. In whole plant experiments, larvae are confined to a single leaf which is eventually detached from the plant and assayed for gene expression. If the order of excision is reversed, from last in whole plant studies, to first in the detached study, the feeding experiment is simplified.
Solanum tuberosum var. Kennebec is propagated by nodal transfer in a simple tissue culture medium and transferred to soil for further growth if desired. Leaves are excised from the parent plant and relocated to Petri dishes where the feeding assay is conducted with the larval stages of M. sexta. Damaged leaf tissue is assayed for the expression of relatively early events in signal transduction. Gene expression analysis identified infestation-specific Cys2-His2 (C2H2) transcription factors, confirming the success of using detached leaves in early response studies. The method is easier to perform than whole plant infestations and uses less space.

The presented method creates natural herbivore damaged plant tissue through the application of Manduca sexta larvae to detached leaves of potato. The plant tissue is assayed for expression of six transcription factor homologs involved in early responses to insect herbivory.

분리 된 잎 어설아는 곤충 만두카 sexta를 츄잉에 의해 감염 하는 동안 감자에 유전자 발현 연구를 단순화 하기 위해

The multitrophic nature of gene expression studies of insect herbivory demands large numbers of biological replicates, creating the need for simpler, more ...

Implantation of a New Micro Acoustic Tag in Juvenile Pacific Lamprey and American Eel

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology has been identified as a way to obtain more detailed information on movement and behavior across a broader spatial scale than is possible with other known technology. The purpose of this method is to provide a detailed step by step instruction on tag implantation for both lampreys and eel. For laboratory studies using actively migrating juvenile Pacific Lamprey (120-160 mm), we determined that the presence of the tag did not alter the swimming ability between tagged and untagged Individuals or have any significant tag loss (&lt;3%). Similar results were determined during laboratory testing of Yellow phase American Eels (113-175 mm). No mortality occurred during a 38-day holding period and there was minimal tag loss (3.8%). The presence of the tag did not have any significant effect on the swimming ability or survival of tagged eels compared to untagged controls and there was minimal tag loss.

This article illustrates the procedure for implanting a micro acoustic tag in juvenile Pacific Lamprey and American Eel. Our application is based on laboratory trials, which indicate that acoustic tags can be successfully implanted with no apparent effects on swimming ability or survival and minimal tag loss.

청소년 태평양 심고와 미국 장 어에 새로운 마이크로 음향 태그의 주입

Juvenile Pacific Lamprey and American eels were used for laboratory evaluations to determine potential effects from tag implantation. Telemetry technology ...