우리는 3D 프린팅 디자인에 그들의 도움에 대한 콜비 카펜터와 스펜서 마티아스 감사합니다. 엘렌 클링거에게 사진 인물 제작에 도움을 주신 것에 대해 감사드리며, 조나단 B. 코흐가 개정에 도움을 준 것에 대해 감사드립니다. 기금은 USDA-ARS 수분 곤충 생물학, 관리 및 체계적인 연구 단위에 의해 제공되었습니다.

1. 꽃가루 트랩 구조물 인쇄
<ol><li>꿀벌이 중첩하고 있는 둥지 상자에 적합한 STL 파일을 다운로드하십시오(예: 바이오베스트 또는 코퍼트 스타일 벌집, https://www.ars.usda.gov/pacific-west-area/logan-ut/pollinating-insect-biology-management-systematics-research/docs/pollen-traps/). 파일은 일반인에게 무료로 제공되며 최종 사용자가 다운로드하고 수정할 수 있습니다.</li>
	<li>프린터 프로그램에서 STL 파일을 엽니다. 프린터 제조업체 의 지시에 따라 4개의 트랩 구성 요소를 빌드합니다. 참고: 트랩 본체가 인쇄할 수 있도록 약 3시간, 캐치 분지가 인쇄할 수 있도록 2시간, 필터및 트랩 클로저 인서트가 인쇄할 수 있는 경우 각각 30분이 인쇄됩니다. 트랩 바디 사이즈는 6cm x 3.8 cm x 7cm입니다.</li>
</ol>2. 꽃가루 트랩 조립
<ol><li>트랩 본체로 인쇄된 지지 구조를 제거하고 트랩 본체의 체 구조를 포함한 유역을 잡는다(도1).</li>
	<li>3/16in. (0.476cm) 드릴 비트를 사용하여 꽃가루 필터의 상승 된 가장자리를 가로지르는 플라스틱 가닥을 지우면 꿀벌이 필터 구멍을 통과하는 것을 방해 할 수 있습니다. 면도날과 사포를 자르는 상자를 사용하여 꽃가루 필터의 평평한 면에 있는 충돌이나 상승 된 가장자리를 고르게하십시오.</li>
	<li>트랩 바디의 한쪽을 통해 플라스틱 필터를 부드럽게 밀어 트랩 본체에 꽃가루 필터를 놓습니다. 트랩의 왼쪽에 커진 필터 콘의 통과를 수용할 수 있는 더 큰 개구부를 가지므로 필터는 한 방향으로만 맞습니다.<ol>
<li>측면 슬릿이 너무 작아서 꽃가루 필터가 매끄럽게 미끄러지기에는 너무 작으면 트랩 본체의 슬릿에서 충분한 플라스틱을 면도기 또는 한 번에 플라스틱의 작은 부분을 제거할 수 있는 다른 도구로 긁어냅니다. 꽃가루 필터와 트랩 바디 사이에 2mm 간격이 없는 곳에 꽃가루 필터가 단단히 고정되어 있는지 확인합니다.</li>
	</ol>
</li>
	<li>트랩 본체 하단의 상승 된 가장자리를 캐치 분지 의 상단의 홈으로 밀어 트랩 본체에 캐치 분지를 부착합니다. 캐치 분지는 트랩 본체의 체 영역 바로 아래에 위치해야한다(그림 1A\u2012E).</li>
	<li>플라스틱을 절단하거나 샌드링하여 트랩 바디에서 캐치 분지를 매끄럽게 배치하고 제거할 수 있도록 적절한 수정을 하십시오. 캐치 분지의 배치는 꽃가루 필터를 제자리에 고정시키고 캐치 분지가 제거 될 때까지 제거 할 수 없으며 캐치 분지가 있는 동안 꽃가루 필터를 삽입 할 수 없습니다. 참고: 여러 개의 두드러기가 서로 가까이 배치되면 각 하이브에 트랩 바디의 고유한 색상 조합을 제공하면 다양한 방향을 가진 두드러기를 배치하는 것 외에도 유역및 꽃가루 필터 구조를 사용하여 돌아오는 작업자가 둥지를 찾는 데 도움이 됩니다.</li>
</ol>3. 범블 꿀벌 식민지 준비
<ol><li>식민지를 배포하기 전에 꽃가루 먹이 24\u201248 h를 중지합니다. 이로 인해 작업자는 저장된 꽃가루를 사용하고 꽃가루를 찾기 위해 둥지를 떠나도록 자극합니다.</li>
	<li>트랩을 설치하는 동안 꿀벌이 탈출하지 못하도록 필터 슬롯에 트랩 클로저 삽입을 삽입하여 설치를 위해 벌집 트랩 본체를 준비합니다.</li>
	<li>꿀벌이 비행하는 것을 방지하기 위해 빨간 불빛 아래에서 작업하여 골판지 외부 상자에서 플라스틱 둥지 상자를 들어 올립니다.</li>
	<li>둥지 앞면에 플라스틱 둥지 입구를 찾습니다. 둥지 공급 업체에 따라 입구의 두 가지 스타일이 있습니다 : Koppert 스타일의 상자 (단계 3.5) 및 Biobest 스타일의 상자 (단계 3.6).</li>
	<li>Koppert 스타일의 하이브 입구의 경우, 두 입구 구멍이 열릴 때까지 입구 탭에 당겨 둥지 입구에 꽃가루 트랩을 장착합니다.<ol>
<li>꽃가루 트랩의 두 튜브를 입구 구멍에 삽입하여 꽃가루 트랩의 체가 바닥에 있는지 확인합니다. 플라스틱 입구 탭을 부드럽게 밀어 꽃가루 트랩을 제자리에 고정시하십시오.</li>
	</ol>
</li>
	<li>Biobest 스타일의 하이브 입구에 꽃가루 트랩을 설치하려면 플랫 헤드 드라이버를 사용하여 둥지 상자에서 플라스틱 입구 장치를 부드럽게 캐고 있습니다. 꽃가루 트랩이 둥지에 단단히 맞을 때까지 둥지 입구 구멍에 꽃가루 트랩을 삽입합니다(도1E).<ol>
<li>필요한 경우 트랩이 둥지 상자에 닿는 테이프 또는 빠른 건조 접착제를 사용하여 트랩을 둥지에 고정합니다.</li>
	</ol>
</li>
	<li>플라스틱 둥지 상자를 골판지 상자에 반환합니다. 골판지는 꽃가루 트랩을 수용하기 위해 잘라야 할 수도 있습니다.</li>
</ol>4. 둥지 배치
<ol><li>둥지 상자를 스터디 영역에 배치합니다. 이 수집 꽃가루의 품질과 수량에 부정적인 영향을 미칠 수 있기 때문에 바람에 대한 강수량과 앵커리지로부터 보호하기 위해 커버를 제공합니다. 과열을 줄이기 위해 적절한 태양 덮개를 온실에 배치 된 두드러기를 제공합니다.</li>
	<li>트랩 바디에서 트랩 클로저 인서트를 제거하여 꿀벌이 자유롭게 사료를 사료할 수 있도록 하여 사료가 주변 지역과 둥지의 위치를 지향할 수 있도록 합니다. 방향 비행 시간은 정상적인 조건에서 24시간 이내에 완료되어야 합니다.</li>
</ol>5. 꽃가루 컬렉션
<ol><li>트랩을 참여시키기 위해 꽃가루 필터를 필터 슬롯에 밀어 고정되도록 합니다.</li>
	<li>캐치 분지를 전면에서 트랩 본체에 슬라이딩하여 완전히 닫을 때까지 설치합니다. 캐치 분지가 지나치게 느슨하거나 트랩 본체에서 떨어지면 고무 밴드를 사용하여 트랩 본체에 고정하십시오.</li>
	<li>꽃가루 필터 구멍이 꿀벌을 수용 할 수있을만큼 큰 지 확인하기 위해 첫 번째 배포에서 꽃가루 트랩에 들어가 고 나가는 꿀벌을 관찰하십시오.<ol>
<li>작업자가 꽃가루 필터를 통과할 수 없는 경우 필터를 제거한 다음 3/16(0.476cm)보다 큰 드릴 비트를 사용하여 구멍 크기를 늘립니다. 너무 큰 구멍이 꽃가루를 수집하지 않기 때문에, 매번 구멍 직경 1/32 (0.079cm)를 증가순차적으로 수행한다.</li>
		<li>꿀벌이 필터를 통과 할 수 있게되면, 다시 들어갈 때 꽃가루가 제거되는지 확인하기 위해 입구를 계속 관찰하십시오. 참고 : 꿀벌은 꽃가루 필터 구멍, 특히 처음 몇 번 통과하지만 이동하는 데 어려움이 있어야합니다. 너무 쉽게 통과하면 꽃가루가 코르비쿨라에서 빠지지 않을 수 있습니다.</li>
	</ol>
</li>
	<li>지정된 꽃가루 수집 기간 이후에 트랩 본체에서 캐치 분지를 밀어 제거합니다.</li>
	<li>실험 설계에 따라 꽃가루 하중을 처리합니다.</li>
	<li>꽃가루 필터를 제거하여 작업자가 다음 꽃가루 수집 기간까지 자유롭게 사료를 사료할 수 있도록 합니다. 트랩 본체는 실험 기간 동안 벌집에 부착된 상태로 남아 있을 수 있다. 참고: 꽃가루 트랩은 연구원이 식민지에서 꽃가루를 수집하고자하는 한 종사 할 수 있습니다. 그러나, 일주일에 24 시간 이상 꽃가루 트랩의 배포는 하이브및 지체 식민지 개발에 무리의 굶주림을 초래할 수 있습니다.</li>
</ol>

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T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+nest+of+Bombus+huntii+Greene+(Hymenoptera:+Apidae).">A nest of Bombus huntii Greene (Hymenoptera: Apidae).</a> Entomological News. 70, 179-182 (1959).</li><li>Husband, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observation+on+colony+of+bumblebee+species+(Bombus+spp).">Observation on colony of bumblebee species (Bombus spp).</a> Great Lakes Entomologist. 10, 83-85 (1977).</li><li>Buttermore, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Observations+of+successful+Bombus+terrestris++(L),+(Hymenoptera:+Apidae)+colonies+in+Southern+Tasmania.">Observations of successful Bombus terrestris (L), (Hymenoptera: Apidae) colonies in Southern Tasmania.</a> Australian Journal of Entomology. 36, 251-254 (1997).</li><li>Goulson, D., Peat, J., Stout, J. C., Tucker, J., Darvill, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Can+alloethism+in+workers+of+the+bumblebee,+Bombus+terrestris,+be+explained+in+terms+of+foraging+efficiency.">Can alloethism in workers of the bumblebee, Bombus terrestris, be explained in terms of foraging efficiency.</a> Animal Behaviour. 64 (1), 123-130 (2002).</li><li>Couvillon, M. J., Jandt, J. M., Duong, N. H. I., Dornhaus, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ontogeny+of+worker+body+size+distribution+in+bumble+bee+(Bombus+impatiens)+colonies.">Ontogeny of worker body size distribution in bumble bee (Bombus impatiens) colonies.</a> Ecological Entomology. 35 (4), 424-435 (2010).</li><li>Russell, A. L., Morrison, S. J., Moschonas, E. H., Papaj, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+pollen+and+nectar+foraging+specialization+by+bumblebees+over+multiple+timescales+using+RFID.">Patterns of pollen and nectar foraging specialization by bumblebees over multiple timescales using RFID.</a> Scientific Reports. 7 (1), 1-13 (2017).</li><li>Hagbery, J., Nieh, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+lifetime+pollen+and+nectar+foraging+preferences+in+bumble+bees.">Individual lifetime pollen and nectar foraging preferences in bumble bees.</a> Naturwissenschaften. 99 (10), 821-832 (2012).</li><li>Baur, A., Strange, J. P., Koch, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Foraging+economics+of+the+Hunt+bumble+bee,+a+viable+pollinator+for+commercial+agriculture.">Foraging economics of the Hunt bumble bee, a viable pollinator for commercial agriculture.</a> Environmental Entomology. 48 (4), 799-806 (2019).</li><li>Winter, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Importation+of+non-native+bumble+bees+into+North+America:+potential+consequences+of+using+Bombus+terrestris+and+other+non-native+bumble+bees+for+greenhouse+crop+pollination+in+Canada,+Mexico,+and+the+United+States.">Importation of non-native bumble bees into North America: potential consequences of using Bombus terrestris and other non-native bumble bees for greenhouse crop pollination in Canada, Mexico, and the United States.</a> San Francisco. 33, (2006).</li><li>Ruz, L., Herrera, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preliminary+observations+on+foraging+activities+of+Bombus+dahlbomii+and+Bombus+terrestris+(Hym:+Apidae)+on+native+and+non-native+vegetation+in+Chile.">Preliminary observations on foraging activities of Bombus dahlbomii and Bombus terrestris (Hym: Apidae) on native and non-native vegetation in Chile.</a> Acta Horticulturae. 561, 165-169 (2001).</li></ol>

저자는 공개 할 것이 없습니다.

꿀벌 식민지 입구에서 꽃가루의 컬렉션은 생태 및 농업 연구의 다양한 수 있습니다. 꿀벌이 꽃가루를 수집하는 꽃원천을 식별하는 것은 식민지의 전반적인 식단19에기여하는 식물의 다양성에 대한 귀중한 정보와 통찰력을 제공합니다. 꽃가루 소스를 식별하는 것은 농업 생산과 야생 땅에서 생태계 서비스의 연구 모두에 영향을 미칩니다12,<sup class="xref"...

범블 꿀벌(봄버스 스PP.) 세계의 온대, 고산, 북극 지역에서 발견되는 큰 강력한곤충이다 1. 그들은 지역 사회를 심고 그들이 방문하는 농업 작물에 대한 중요한 수분 서비스를 제공하는 것이 중요합니다2. 최근 여러 종의 풍요로움과 분포가 감소하면서 수분공급자가 대중의 인식3의최전선에 서게 되면서 그 중요성이 제기되고 있다. 연구원은 꿀벌 사료4에다양하고 풍부한 꽃 자원의 부족을 포함하여 인구 감소에 기여할 가능성이 있는 몇몇 스트레스를 확인했습니다. 어떤 식물 종 범블 꿀벌 사료에서 식별하면 연구원과 토지 관리자는 꿀벌이 자원 가용성, 경쟁 및 인위적장애의변화에 어떻게 반응할 수 있는지 이해할 수 있습니다5,6.
꿀벌의 꽃가루 를 먹는 선호도를 조사하는 연구는 종종 꽃에서 먹는 개별 꿀벌을 잡은 다음 추가 처리 및식별을위해 표본에서 관상 꽃가루 하중을 제거한 연구원에 의해 수행됩니다7,8,9,10. 이 방법은 꿀벌 종의 종 또는 조립이 영역7의자원을 활용하는 방법에 대한 통찰력을 제공하지만, 벌집 사이의 선호도에 대한 집중적이고 잠재적 인 차이는 사료 꿀벌(11)의기원의 식민지를 식별하기 위해 추가 분자 분석없이 분별 할 수 없다.
역학을 포기하는 일부 연구에 대 한, 개별 식민지에서 연구를 수행 하는 것이 좋습니다.; 그러나 야생 범블 꿀벌 둥지는 일반적으로 지하 또는 지상층에 위치하여12를찾기가 어렵습니다. 상업적으로 생산된 범블 벌두집은 연구원에게 더 큰 접근성과 더 나은 실험 적 제어를 제공하고 작업자로부터 꽃가루제거는 여전히 주로 포집포로 돌아가서 혈관 꽃가루 하중13,14를수동으로 제거하여 수행된다. 꿀벌의 코르비쿨라로부터 수작업으로 꽃가루를 제거하는 것은 꽃가루 사료를 반환하는 비율이 낮을 수 있는 벌집 입구에서 특히 시간당 낮은 꽃가루 수율로 시간이 집중적입니다. 또한 꿀벌에서 꽃가루를 수동으로 제거하면 방해받은 작업자의 찌르기가 발생할 수 있습니다.
꽃가루 트랩은 수십 년 동안 꿀벌에서 꽃가루를 실험제거하는 데 사용되어 왔다15; 그러나 꿀벌에서 꽃가루를 제거하는 수동 적인 방법은 개발되지 않았습니다. 꽃가루를 반납하지 못하도록 하는 메커니즘을 개발하는 데 주요 장애물은 꿀벌서식지(16)에존재하는 작업자 크기의 큰 변화이다. 꿀벌 꽃가루 트랩은 꿀벌 노동자 크기가 별로 변하지 않기 때문에 크게 효과적입니다. 또한,이 함정은 설치 후 사소한 조작을 필요로하고 꿀벌을 희생 할 필요가 없습니다17. 이것은 그들이 하이브로 돌아갈 때 노동자의 뒷다리에서 꽃가루를 제거하는 스크린 이나 플라스틱 표면을 사용하여 달성된다. 이 트랩은 꽃가루 하중의 일부만 제거하여 사료를 반환하고 꽃가루 컬렉션에서 다양한 효율성을 초래하는 다양한 디자인의 설계를 제거합니다. 꽃가루가 꿀벌 다리에서 제거되면 화면을 통해 꿀벌이 접근 할 수없는 수집 분지로 떨어지므로 연구원은 벌집에 약간의 방해만으로 제거 할 수 있습니다.
본 연구의 목적은 꿀벌 벌집에서 꽃가루를 수집하는 데 사용되는 기술을 조정하고 3D 인쇄 구조를 사용하여 꿀벌 둥지에 적용하고 봄부스 헌티의식민지에 트랩 디자인을 테스트하는 것입니다. 설계 과정은 트랩이 생산 저렴해야한다는 가정을 따라, 꿀벌 종의 다양한 적응, 꿀벌에 최소한의 해또는 방해를 야기, 꽃가루 제거의 속도는 꽃가루의 손 수집을 초과해야한다는 가정. 3차원 인쇄 기술은 다재다능하고 쉽게 접근할 수 있으며, 연구자가 특정 목적을 위해 객체를 복제하고 수정할 수 있는 비용 효율적인도구입니다(18). 여기에 제시된 기술은 사용자에게 꽃가루 트랩을 구축하고 시판되는 범블 꿀벌 식민지에 부착하도록 지시합니다. 트랩은 야생 식민지와 함께 사용하도록 설계되지 않았습니다. 이 함정은 둥지 상자로 돌아갈 때 꿀벌을 운반하는 꽃가루의 뒷다리에서 관두 꽃가루 하중을 수동적으로 제거합니다.

<table><tbody><tr><td>MakerBot Replicator+</td><td>MakerBot</td><td>Model PABH65</td><td></td></tr><tr><td>MakerBot Tough Material</td><td>PLA Filament</td><td></td><td>various colors</td></tr><tr><td>Nest Box</td><td>Biobest</td><td></td><td>Not sold publicly without bee purchase</td></tr></tbody></table>

a 3d printed pollen trap for bumble bee (bombus) hive entrances

꽃가루에 꿀벌 사료가 있는 식물 원을 확인하려면 분석을 위해 관상 꽃가루 하중을 제거하기 위해 개인을 수집해야합니다. 이것은 전통적으로 둥지 입구 또는 꽃에 사료를 그물, 얼음에 꿀벌을 냉각, 다음 집게 또는 브러시로 코르비쿨라에서 꽃가루 부하를 제거에 의해 수행되었습니다. 이 방법은 시간과 노동 집약적이며 정상적인 위조 동작을 변경할 수 있으며 작업을 수행하는 작업자의 쏘는 사고가 발생할 수 있습니다. 꿀벌 벌집에 사용되는 꽃가루 트랩은 둥지 입구의 화면을 통과할 때 작업자의 다리에서 코비알 꽃가루하를 분리하여 꽃가루를 수집합니다. 트랩은 최소한의 노동력으로 포어 꿀벌을 반환에서 꽃가루의 대량을 제거 할 수 있습니다, 아직 현재까지 범블 꿀벌 식민지와 함께 사용할 수있는 이러한 함정을 사용할 수 없습니다. 꿀벌 식민지 내의 작업자는 크기가 다를 수 있으므로 이 메커니즘을 상업적으로 키서 꿀벌 벌집에 적응하기 어려운 입구의 크기 선택이 될 수 있습니다. 3D 프린팅 설계 프로그램을 사용하여, 우리는 성공적으로 반품 꿀벌 사료의 다리에서 관상 꽃가루 부하를 제거하는 꽃가루 트랩을 만들었습니다. 이 방법은 연구원이 식민지로 돌아오는 범블 꿀벌 사료에서 꽃가루를 수집하는 데 필요한 시간을 크게 줄입니다. 우리는 설계, 꽃가루 제거 효율 테스트의 결과를 제시하고, 다양한 범블 꿀벌 종 또는 둥지 상자 디자인에 트랩을 적응하기 위해 조사관을위한 수정 영역을 제안한다.

8가지 꽃가루 필터 설계는 꿀벌 작업자를 반환하는 코비큘러 꽃가루 하중을 제거하는 데 효능과 효율성을 결정하기 위해 테스트되었습니다. 모든 설계는 돌아오는 포어에서 적어도 하나의 코비큘러 꽃가루 부하를 제거하는 데 성공했습니다. 그러나, 일부는 부드러름을 떠나거나 입력에서 노동자를 느리게하거나 꽃가루 부하를 제거하지 못하는 것으로 나타났다(표 1). 다양한 필터가 ...

Watch this Scientific Journal Video about A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances at JoVE.com

A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances

우리는 벌집으로 돌아오는 범블 꿀벌(봄푸스)근로자로부터 꽃가루를 수집하는 비 치명적이고 자동화 된 메커니즘을 제시합니다. 장치 생산, 준비, 설치 및 사용에 대한 지침이 포함되어 있습니다. 3D 프인쇄 객체를 사용하여 설계를 수정하는 것은 시기적이고 효율적이며 테스트를 위한 빠른 처리를 허용했습니다.

범블 비(봄푸스)하이브 입구를위한 3D 인쇄 꽃가루 트랩

To verify the plant sources from which bumble bees forage for pollen, individuals must be collected to remove their corbicular pollen loads for analysis.  ...

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A 3D Printed Pollen Trap for Bumble Bee (Bombus) Hive Entrances

5.1K 조회수.  Utah State University. 우리는 벌집으로 돌아오는 범블 꿀벌(봄푸스)근로자로부터 꽃가루를 수집하는 비 치명적이고 자동화 된 메커니즘을 제시합니다. 장치 생산, 준비, 설치 및 사용에 대한 지침이 포함되어 있습니다. 3D 프인쇄 객체를 사용하여 설계를 수정하는 것은 시기적이고 효율적이며 테스트를 위한 빠른 처리를 허용했습니다.

비디오: A 3D Printed Pollen Trap for Bumble Bee Bombus Hive Entrances

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the survival of a honeybee colony depends on the exploitation of food sources, forager bees learn and memorize variable flower sites as well as their profitability. Forager bees can be easily trained in natural settings where they forage at a feeding site and learn the related signals such as odor or color. Appetitive associative learning can also be studied under controlled conditions in the laboratory by conditioning the proboscis extension response (PER) of individually harnessed honeybees3,4. This learning paradigm enables the study of the neuronal and molecular mechanisms that underlie learning and memory formation in a simple and highly reliable way5-12. A behavioral pharmacology approach is used to study molecular mechanisms. Drugs are injected systemically to interfere with the function of specific molecules during or after learning and memory formation13-16.
Here we demonstrate how to train harnessed honeybees in PER conditioning and how to apply drugs systemically by injection into the bee flight muscle.

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees Apis mellifera

We demonstrate how to implement a behavioral pharmacology method in an appetitive olfactory conditioning paradigm in honeybees (Apis mellifera) by systemic application of drugs. This method allows investigation of the mechanisms underlying learning and memory formation in a simple and reliable way.

꿀벌의 코 확장 응답 클래식 컨디셔닝에 Behavioural 약리학 ( 아피스 mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the ...

연구

신경과학

A Push-pull Protocol to Reduce Colonization of Bird Nest Boxes by Honey Bees

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. These bees select cavities that are suitable nest sites for birds, resulting in competition. The difficulty of removing bees and their defensive behavior makes a prevention protocol necessary. Here, we describe a push-pull integrated pest management protocol to deter bees from inhabiting bird boxes by applying a bird safe insecticide, permethrin, to repel bees from nest boxes, while simultaneously attracting them to pheromone-baited swarm traps. Shown here is an example experiment using Barn Owl nest boxes. This protocol successfully reduced colonization of Barn Owl nest boxes by Africanized honey bees. This protocol is flexible, allowing adjustments to accommodate a wide range of bird species and habitats. This protocol could benefit conservation efforts where AHB are located.

Preventing colonization of bird nest boxes by invasive Africanized honey bees is important for conservation efforts of nest-site limited birds. We provide an integrated pest management approach to &#34;push&#34; bees away from nest boxes with a repellant insecticide, permethrin, and &#34;pull&#34; them toward pheromone baited swarm traps.

푸시 풀 프로토콜 꿀 꿀벌에 의해 식민 새의 둥지 상자를 감소시키는

Introduction of the invasive Africanized honey bee (AHB) into the Neotropics is a serious problem for many cavity nesting birds, specifically parrots. ...

환경과학

Empirical, Metagenomic, and Computational Techniques Illuminate the Mechanisms by which Fungicides Compromise Bee Health

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered &#34;bee-safe,&#34; there is mounting evidence that fungicide residues in pollen are associated with bee declines (for both honey and bumble bee species). While the mechanisms remain relatively unknown, researchers have speculated that bee-microbe symbioses are involved. Microbes play a pivotal role in the preservation and/or processing of pollen, which serves as nutrition for larval bees. By altering the microbial community, it is likely that fungicides disrupt these microbe-mediated services, and thereby compromise bee health. This manuscript describes the protocols used to investigate the indirect mechanism(s) by which fungicides may be causing colony decline. Cage experiments exposing bees to fungicide-treated flowers have already provided the first evidence that fungicides cause profound colony losses in a native bumble bee (Bombus impatiens). Using field-relevant doses of fungicides, a series of experiments have been developed to provide a finer description of microbial community dynamics of fungicide-exposed pollen. Shifts in the structural composition of fungal and bacterial assemblages within the pollen microbiome are investigated by next-generation sequencing and metagenomic analysis. Experiments developed herein have been designed to provide a mechanistic understanding of how fungicides affect the microbiome of pollen-provisions. Ultimately, these findings should shed light on the indirect pathway through which fungicides may be causing colony declines.

Microbial consortia within bumble bee hives enrich and preserve pollen for bee larvae. Using next generation sequencing, along with laboratory and field-based experiments, this manuscript describes protocols used to test the hypothesis that fungicide residues alter the pollen microbiome, and colony demographics, ultimately leading to colony loss.

실험, Metagenomic, 및 계산 기법 밝히는 어떤 균 타협 꿀벌 건강 메커니즘

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered ...

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 ...

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings (GCMR) in the Insect Antennal Lobe

All organisms inhabit a world full of sensory stimuli that determine their behavioral and physiological response to their environment. Olfaction is especially important in insects, which use their olfactory systems to respond to, and discriminate amongst, complex odor stimuli. These odors elicit behaviors that mediate processes such as reproduction and habitat selection1-3. Additionally, chemical sensing by insects mediates behaviors that are highly significant for agriculture and human health, including pollination4-6, herbivory of food crops7, and transmission of disease8,9. Identification of olfactory signals and their role in insect behavior is thus important for understanding both ecological processes and human food resources and well-being.
	To date, the identification of volatiles that drive insect behavior has been difficult and often tedious. Current techniques include gas chromatography-coupled electroantennogram recording (GC-EAG), and gas chromatography-coupled single sensillum recordings (GC-SSR)10-12. These techniques proved to be vital in the identification of bioactive compounds. We have developed a method that uses gas chromatography coupled to multi-channel electrophysiological recordings (termed 'GCMR') from neurons in the antennal lobe (AL; the insect's primary olfactory center)13,14. This state-of-the-art technique allows us to probe how odor information is represented in the insect brain. Moreover, because neural responses to odors at this level of olfactory processing are highly sensitive owing to the degree of convergence of the antenna's receptor neurons into AL neurons, AL recordings will allow the detection of active constituents of natural odors efficiently and with high sensitivity. Here we describe GCMR and give an example of its use.
	Several general steps are involved in the detection of bioactive volatiles and insect response. Volatiles first need to be collected from sources of interest (in this example we use flowers from the genus Mimulus (Phyrmaceae)) and characterized as needed using standard GC-MS techniques14-16. Insects are prepared for study using minimal dissection, after which a recording electrode is inserted into the antennal lobe and multi-channel neural recording begins. Post-processing of the neural data then reveals which particular odorants cause significant neural responses by the insect nervous system.
	Although the example we present here is specific to pollination studies, GCMR can be expanded to a wide range of study organisms and volatile sources. For instance, this method can be used in the identification of odorants attracting or repelling vector insects and crop pests. Moreover, GCMR can also be used to identify attractants for beneficial insects, such as pollinators. The technique may be expanded to non-insect subjects as well.

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings GCMR in the Insect Antennal Lobe

Olfactory cues mediate many different behaviors in insects, and are often complex mixtures comprised of tens to hundreds of volatile compounds. Using gas chromatography with multi-channel recording in the insect antennal lobe, we describe a method for the identification of bioactive compounds.

곤충 Antennal 엽에 가스 크로마토 그래피 - 멀티 유닛 레코딩 (GCMR)를 사용하여 강한 휘발성 물질의 식별

All  organisms inhabit a world full of sensory stimuli that determine their  behavioral and physiological response to their environment. Olfaction is ...

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees (Apis mellifera L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker bees spend the first half of their life inside the dark hive, where they are nursing the larvae or building the regular hexagonal combs for food (e.g. pollen or nectar) and brood3. The antennae are extraordinary multisensory feelers and play a pivotal role in various tactile mediated tasks4, including hive building5 and pattern recognition6. Later in life, each single bee leaves the hive to forage for food. Then a bee has to learn to discriminate profitable food sources, memorize their location, and communicate it to its nest mates7. Bees use different floral signals like colors or odors7, 8, but also tactile cues from the petal surface9 to form multisensory memories of the food source. Under laboratory conditions, bees can be trained in an appetitive learning paradigm to discriminate tactile object features, such as edges or grooves with their antennae10, 11, 12, 13. This learning paradigm is closely related to the classical olfactory conditioning of the proboscis extension response (PER) in harnessed bees14. The advantage of the tactile learning paradigm in the laboratory is the possibility of combining behavioral experiments on learning with various physiological measurements, including the analysis of the antennal movement pattern.

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

In this protocol we show how to condition harnessed honey bees to tactile stimuli and introduce a 2D motion capture technique for analyzing the kinematics of fine-scale antennal sampling pattern.

촉각 시설과 꿀벌에 Antennal 샘플링 전략의 움직임 분석 (<em&gt; API를 mellifera</em&gt; L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker ...

A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). There are several behavioral mechanisms through which an insect learns salient associations and relates them to these events. It is important to understand this behavioral plasticity for programs aimed toward assisting insects that are beneficial for agriculture. This understanding can also be used for discovering solutions to biomedical and agricultural problems created by insects that act as disease vectors and pests. The Proboscis Extension Response (PER) conditioning protocol was developed for honey bees (Apis mellifera) over 50 years ago to study how they perceive and learn about floral odors, which signal the nectar and pollen resources a colony needs for survival. The PER procedure provides a robust and easy-to-employ framework for studying several different ecologically relevant mechanisms of behavioral plasticity. It is easily adaptable for use with several other insect species and other behavioral reflexes. These protocols can be readily employed in conjunction with various means for monitoring neural activity in the CNS via electrophysiology or bioimaging, or for manipulating targeted neuromodulatory pathways. It is a robust assay for rapidly detecting sub-lethal effects on behavior caused by environmental stressors, toxins or pesticides.
We show how the PER protocol is straightforward to implement using two procedures. One is suitable as a laboratory exercise for students or for quick assays of the effect of an experimental treatment. The other provides more thorough control of variables, which is important for studies of behavioral conditioning. We show how several measures for the behavioral response ranging from binary yes/no to more continuous variable like latency and duration of proboscis extension can be used to test hypotheses. And, we discuss some pitfalls that researchers commonly encounter when they use the procedure for the first time.

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

코 확장 응답 곤충의 행동 소성을 조사하기위한 프로토콜 : 기본, 바이오 메디컬에 응용 프로그램, 및 농업 연구

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). ...

Radio Frequency Identification and Motion-sensitive Video Efficiently Automate Recording of Unrewarded Choice Behavior by Bumblebees

We present two methods for observing bumblebee choice behavior in an enclosed testing space. The first method consists of Radio Frequency Identification (RFID) readers built into artificial flowers that display various visual cues, and RFID tags (i.e., passive transponders) glued to the thorax of bumblebee workers. The novelty in our implementation is that RFID readers are built directly into artificial flowers that are capable of displaying several distinct visual properties such as color, pattern type, spatial frequency (i.e., &#8220;busyness&#8221; of the pattern), and symmetry (spatial frequency and symmetry were not manipulated in this experiment). Additionally, these visual displays in conjunction with the automated systems are capable of recording unrewarded and untrained choice behavior. The second method consists of recording choice behavior at artificial flowers using motion-sensitive high-definition camcorders. Bumblebees have number tags glued to their thoraces for unique identification. The advantage in this implementation over RFID is that in addition to observing landing behavior, alternate measures of preference such as hovering and antennation may also be observed. Both automation methods increase experimental control, and internal validity by allowing larger scale studies that take into account individual differences. External validity is also improved because bees can freely enter and exit the testing environment without constraints such as the availability of a research assistant on-site. Compared to human observation in real time, the automated methods are more cost-effective and possibly less error-prone.

This video describes Radio-Frequency Identification (RFID) and motion-sensitive video recording methods to monitor choice behavior by bumblebees.

무선 주파수 식별하고 효율적으로 모션에 민감한 비디오 땅벌에 의해 무보수 선택 행동의 기록을 자동화

We present two methods for observing bumblebee choice behavior in an enclosed testing space. The first method consists of Radio Frequency Identification ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from flowering plants. Only a few studies have described the gustatory responses of bees toward toxins in food, and these experiments have mainly used the proboscis extension response on restrained honey bees. Here, a new behavioral assay is presented for measuring the feeding responses of freely-moving, individual worker bumble bees to nutrients and toxins. This assay measures the amount of solution ingested by each bumble bee and identifies how tastants in food influence the microstructure of the feeding behavior.
The solutions are presented in a microcapillary tube to individual bumble bees that have been previously starved for 2-4 hr. The behavior is captured on digital video. The fine structure of the feeding behavior is analyzed by continuously scoring the position of the proboscis (mouthparts) from video recordings using event logging software. The position of the proboscis is defined by three different behavioral categories: (1) proboscis is extended and in contact with the solution, (2) proboscis is extended but not in contact with the solution and (3) proboscis is stowed under the head. Furthermore the speed of the proboscis retracting away from the solution is also estimated.
In the present assay the volume of solution consumed, the number of feeding bouts, the duration of the feeding bouts and the speed of the proboscis retraction after the first contact is used to evaluate the phagostimulatory or the deterrent activity of the compounds tested.
This new taste assay will allow researchers to measure how compounds found in nectar influence the feeding behavior of bees and will also be useful to pollination biologists, toxicologists and neuroethologists studying the bumble bee&#39;s taste system.

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A novel behavioral assay is described for investigating the short term gustatory responses of the mouthparts of freely-moving bumble bees (Bombus terrestris) toward nutrients and toxins in solution.

소설 행동 분석은 (엉망으로 꿀벌을 자유롭게 이동 개인의 미각 응답을 조사하기<em&gt; 뒤의 terrestris</em&gt;)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...

Collection and Identification of Pollen from Honey Bee Colonies

Researchers often collect and analyze corbicular pollen from honey bees to identify the plant sources on which they forage for pollen or to estimate pesticide exposure of bees via pollen. Described herein is an effective pollen-trapping method for collecting corbicular pollen from honey bees returning to their hives. This collection method results in large quantities of corbicular pollen that can be used for research purposes. Honey bees collect pollen from many plant species, but typically visit one species during each collection trip. Therefore, each corbicular pollen pellet predominantly represents one plant species, and each pollen pellet can be described by color. This allows the sorting of samples of corbicular pollen by color to segregate plant sources. Researchers can further classify corbicular pollen by analyzing the morphology of acetolyzed pollen grains for taxonomic identification. These methods are commonly used in studies related to pollinators such as pollination efficiency, pollinator foraging dynamics, diet quality, and diversity. Detailed methodologies are presented for collecting corbicular pollen using pollen traps, sorting pollen by color, and acetolyzing pollen grains. Also presented are results pertaining to the frequency of pellet colors and taxa of corbicular pollen collected from honey bees in five different cropping systems.

We describe methods for collecting corbicular pollen from honey bees as well as protocols for color sorting, acetolysis, and microscope slide preparation of pollen for taxonomic identification. In addition, we present pellet color and taxonomic diversity of corbicular pollen collected from five cropping systems using pollen traps.

꿀벌 식민지에서 꽃가루의 수집 및 식별

Researchers often collect and analyze corbicular pollen from honey bees to identify the plant sources on which they forage for pollen or to estimate ...

범블 비(봄푸스)하이브 입구를위한 3D 인쇄 꽃가루 트랩

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