이 연구는 부여 105AS-13.2.3-BQ-B1 교육부에서 동물의 국과 식물 건강 검사 및 검역, 농업위원회, 집행 위안 그랜트 103-2313-B-197-002-MY3에서에 의해 지원되었다 과학 기술 (MOST)의.

1. 준비 <ol><li> 50 %의 설탕 시럽 1 L합니다. 1 L의 DDH 2 O.에 1kg 자당 녹이고 </li><li> BLD에서 pyriproxyfen (PPN) 솔루션을 준비합니다. 10,000 ppm의 PPN 원액 1.1 L을 4 ° C에서 2 DDH O. 스토어 멸균 1 L에 PPN 100 ㎖ 용액을 희석. </li><li> 다음 실험을위한 BLD 0.1, 1, 10 및 100 ㎎ / ㎏ (PPM)의 최종 농도로 PPN 원액을 희석. </li><li> (식민지 레벨) PPN-시럽을합니다. 다음 실험에 대한 50 %의 설탕 시럽 10 및 100 ppm으로 최종 농도로 PPN 주식을 희석. </li><li> 꿀 꿀벌 사육. 참고 : 여기, 실험 위치는 국립 일란 대학 (NIU) 양봉장, 이순신 - 란시, 대만은; (GPS 좌표 : N24.747278, E121.746200). <ol><li> 꿀벌 (아피스 mellifera이 L.)는 식품 수량 매주 식민지 확인하고 (꿀 저장 영역이 비어) 필요한 경우 1리터 50 %의 설탕 시럽을 공급. 건강한 정의일반적 산란 퀸 각 콜로니의 꿀벌 빗 9 프레임으로서 콜로니. </li></ol></li><li> 기본 애벌레 다이어트 (BLD)의 100 mL로 준비합니다. (DDH 2 O) 멸균 증류수, 탈 이온수에 6 % D- 글루코오스, 프 룩토 오스 6 %, 1 % 효모 추출물을 용해시키고, 50 % 로얄 젤리 보충. 4 ° C에서 보관하지만 3 일 이상합니다. 실험 전에 35 ° C로 사전 따뜻하고 3 일 이내에 사용합니다. </li></ol> 생체 수유 방법 2. 노트 : 생체 내에서 공급 법은 헨리 등의 알로부터 변형되었다. (20) <ol><li> 꿀 꿀벌 유충 선택 및 라벨링. <ol><li> 4 프레임 부에 퀸을 제한하면서 4 프레임 5 개 프레임으로 건강한 콜로니 9 프레임 분할 퀸 excluder입니까. 산란을 위해 4 프레임 섹션에서 적어도 하나 개의 빈 프레임을 남겨주세요. </li><li> 여왕이 1 일 산란 후계란의 외관을 검사 프레임 1 일령 유충이 부화까지 72 시간 (4 일째)에 벌통 내부의 알을 유지한다. 손으로 테스트 하이브 밖으로 (24 시간 내에 부화) 1 일 된 노동자 유충을 포함하는 프레임 중 하나를 가지고 꿀벌 브러시 프레임에서 꿀벌 노동자를 제거합니다. </li><li> 투명 슬라이드 (크기 = 길이 X 너비 X 두께 29.7 mm X 21mm X 0.1 mm)를 가진 프레임을 덮고 압정 (도 1a)와 프레임의 가장자리에 상기 투명 슬라이드 손톱. </li><li> 각각의 치료를위한, 임의로 하나 50 일령 유충 (4 일째)을 선택하고, 상기 투명 슬라이드 영구 마커 펜을 사용하여 각 브루 셀 마크 (도 1a 및도 1b). 참고 : 다른 치료 사이의 혼동을 피하기 위해 영구 마커 펜을 사용하여뿐만 아니라 프레임과 투명 슬라이드의 각 치료의 정보를 작성합니다. 표시된 슬라이드를 제거하고 생체에 대한 유지 &lt;/ EM&gt; 급전 관찰. </li></ol></li><li> 생체 먹이. <ol><li> 의 흡기 양에 따라 각각 10 μL, 10 μL, 20 μL로 일 1, 2, 3에 피펫으로 각 표지 브루 셀 PPN-BLD (0.1, 1, 10, 100 PPM)의 상이한 농도를 추가 유충의 나이. 대조군에 BLD (NO PPN)의 동일한 금액을 추가합니다. 이에 따라, 각 분류 브루 셀 PPN-BLD의 총 투여 량은 4, 40, 400, 4000 페이지에 축적한다. 참고 : 표시된 투명 슬라이드하여 표시 무리 세포를 인식하고 공급 한 후 표시된 투명 슬라이드를 제거합니다. 일벌에 의해 무리 세포의 청소를 방지하기 위해 공급하는 신선한 BLD를 사용합니다. </li><li> 더 관찰의 원래 식민지로 PPN 처리 된 프레임을 돌려줍니다. 참고 : 각각의 치료는 네 개의 식민지에서 네 생물 반복 있습니다. </li></ol></li><li> 7 일에서 처리 유충의 관측. <ol><li> 일을 관찰하려면전자 표지 무리 세포의 사망률과 상한 율을 기록하기 위해 프레임에 표시된 투명 슬라이드를 반환 유충을 PPN은 처리. </li></ol></li><li> 하루에 13에서 처리 번데기와 우화 요금의 관측. <ol><li> 덮인 무리 세포의 꿀벌 왁스를 제거합니다. </li><li> 브루 셀에 소프트 팁 핀셋을 넣고 클램프 번데기를 아주 약간 부드럽게 번데기를 꺼내. </li><li> 각 웰 아래 기관 조직의 이중층으로 24 웰 조직 배양 플레이트에 번데기로 이동. 번데기 전송 중에 손상과 사망률을 기록합니다. </li><li> (약 팔일) 출현 될 때까지 34 ° C, 70 % 상대 습도 인큐베이터에서 24 웰 조직 배양 플레이트 유지. </li><li> 관찰과 번데기와 등장 꿀벌을 기록합니다. </li></ol></li><li> 통계 <ol><li> ± SD 평균값으로 기록 된 데이터와 현재 계산 </li><li> SA에 의해 변동 (ANOVA)을 이용하여 분석 데이터를 분석S 및 다른 처리의 두 방법의 차이를 분석하기 위해 최하위 차 (LSD)를 사용하여 시험. P - 값 &lt;0.05로 통계적으로 유의 한 정의합니다. 테이블의 동일한 열에서 서로 다른 글자는 통계적 분석에 의해 현저한 효과를 나타냈다. </li></ol></li></ol> 벌집에서 식민지 수준에서 PPN 3. 독성 <ol><li> 꿀벌 그룹을 설정합니다. <ol><li> 프레임 4 (파트 A) 및도 5의 프레임 (파트 B) 4 프레임 부에 여왕으로 제한하면서 건강한 콜로니 9 개 프레임을 분할하는 수직 퀸 excluder입니까. 산란을위한 4 프레임 부분에 적어도 하나 개의 빈 프레임을 남겨주세요. 참고 : 다른 여왕 부품 사이에 이동 여왕을 방지하기 위해 여왕 부분의 상단에있는을 excluder 넣습니다. </li><li> 여왕이 1 일 산란 한 후, 달걀 모양의 프레임을 선택합니다. 손으로 A 부분에서 계란을 포함하는 적절한 프레임을 가지고 꿀벌 근로자를 제거꿀벌 브러시로 프레임입니다. </li><li> 투명 슬라이드 프레임을 덮고 압정와 프레임의 가장자리에 상기 투명 슬라이드 손톱. </li><li> 임의로 함유 달걀 100 개 브루 셀을 선택하여 상기 투명 슬라이드 영구 마커를 사용하여 각 브루 셀을 표시한다. 그룹 1. 다른 치료 사이의 혼동을 피하기 위해 영구 마커를 사용하여 프레임과 투명 슬라이드의 각 치료의 정보를 기록 이러한 100 개 무리 세포를 할당합니다. </li><li> 3 일 A 부분에 표시된 프레임을 반환 한 후 알을 낳기 위해 파트 B에 여왕을 전송합니다. </li><li> 일일 후, 파트 B의 프레임을 검사 한 적절한 프레임을 선택하고, 단계 3.1.4에 기술 된 바와 같이 계란 함유 100 브루 셀 라벨. 그룹이 이러한 100 개 무리 세포를 할당합니다. </li><li> 3 일 파트 B로 표시된 프레임을 반환 한 후 알을 낳기에이 부분에 여왕을 전송합니다. </li><li> 6 번 파트 A와 파트 B의 여왕 교환을 반복하고 수치 그룹을 할당,각각 (도 2). 9 개 그룹의 총이 있어야합니다. </li></ol></li><li> 일 13 PPN 설탕 시럽 T reat 꿀벌 콜로니 (도 2). <ol><li> 1 L 플라스틱 꿀벌 급전 박스 (10) 또는 100 PPM (W X L x 높이 = 20 cm X 30cm X 3.5 cm)를 함유하는 50 % PPN 설탕 시럽을 추가 실험 콜로니의 프레임의 상부에 박스를 넣어. 참고 : 무리 세포가 봉인 된 것처럼 그룹 1 13 일째에 PPN을받지 않습니다. </li><li> 단계 2.2에 기재된 바와 같이 1 L 50 % 설탕 시럽 (NO PPN)와 대조군 사료. </li></ol></li><li> 5 일 (그림 2)에서 각 그룹에 대해 100 개 라벨 무리 세포의 계란 부화 속도로 일일 된 유충 기록을 계산합니다. 꿀 꿀벌 계란은 일반적으로 0일 유충으로 부화 3 일이 걸릴; 따라서 확인하고 100 계란 함유 하루 1에 무리 세포를 라벨 및 부화 속도의 비율을 얻기 위해 하루 5에서 각 그룹에 대해 1 일 된 유충의 수를 계산합니다. </li><li> 기음날 (11) (도 2)에서 각 그룹 (100 개) 표지 브루 셀의 애벌레 캐핑 레이트 캡핑 브루 셀 기록 ount. 6 ~ 7 일 된 애벌레 무리 세포는 애벌레 pupating 꿀 꿀벌 노동자 꿀벌 왁스로 제한됩니다. </li><li> 번데기 성숙을 관찰하고 17 일 (도 2)에서 각 그룹 (100 개) 표지 브루 셀의 우화 율을 기록한다. <ol><li> 덮인 무리 세포의 꿀벌 왁스를 제거하고 부드럽게 부드러운 밀고 족집게와 번데기를 꺼내. 각 웰 아래 기관 조직의 이중층으로 24 웰 조직 배양 플레이트에 번데기로 이동. </li><li> 출현 될 때까지 34 ° C, 70 % RH에서 항온 24 웰 조직 배양 판을 유지한다. </li><li> 관찰과 번데기를 기록하고 그룹 9까지 각 그룹 (49 실험 일)를 위해 꿀벌을 등장. 참고 : 각 치료는 네 생물 반복 있습니다. </li></ol></li><li> 통계 <ol><li> 기록 데이터를 계산하고, 평균으로서 본± SD. </li><li> 치료의 쌍 사이의 유의 한 차이를 분석하여 (예를 들어 0 PPM / 10 PPM 10 PPM / 100 ppm을 0 PPM / 100 PPM) 학생의 양측의 t -test를 사용하여 각 그룹이다. P - 값 &lt;0.05처럼 통계적으로 유의 한 정의합니다. </li></ol></li></ol>

<ol>
	<li>Ruffi nengo, S. R., 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=Integrated+pest+management+to+control+Varroa+destructor+and+its+implications+to+Apis+mellifera+colonies.">Integrated pest management to control Varroa destructor and its implications to Apis mellifera colonies.</a> Zootecnia Trop. 32 (2), 149-168 (2014).</li><li>Mullin, C. A., 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=High+levels+of+miticides+and+agrochemicals+in+North+American+apiaries,+implications+for+honey+bee+health.">High levels of miticides and agrochemicals in North American apiaries, implications for honey bee health.</a> PLoS One. 5, e9754 (2010).</li><li>Lu, C. A., Chang, C. H., Tao, L., Chen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distributions+of+neonicotinoid+insecticides+in+the+Commonwealth+of+Massachusetts,+a+temporal+and+spatial+variation+analysis+for+pollen+and+honey+samples.">Distributions of neonicotinoid insecticides in the Commonwealth of Massachusetts, a temporal and spatial variation analysis for pollen and honey samples.</a> Environ. Chem. 13, 4-11 (2016).</li><li>Tsai, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+coupling+the+pesticide+use+reduction+with+environmental+policy+for+agricultural+sustainability+in+Taiwan.">Analysis of coupling the pesticide use reduction with environmental policy for agricultural sustainability in Taiwan.</a> Environ. &amp; Pollut. 2, 59-65 (2013).</li><li>Weng, Z. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pesticide+market+status+and+development+trend+(in+Chinese).">Pesticide market status and development trend (in Chinese).</a> PRIDE. , (2016).</li><li>Kevan, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pollinators+as+bioindicators+of+the+state+of+the+environment,+species,+activity+and+diversity.">Pollinators as bioindicators of the state of the environment, species, activity and diversity.</a> Agric. Ecosys. Environ. 74 (1-3), 373-393 (1999).</li><li>Kevan, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forest+application+of+the+insecticide+fenitrothion+and+its+effect+on+wild+bee+pollinators+(Hymenoptera:+Apoidea)+of+lowbush+blueberries+(Vaccinium+SPP.)+in+Southern+New+Brunswick,+Canada.">Forest application of the insecticide fenitrothion and its effect on wild bee pollinators (Hymenoptera: Apoidea) of lowbush blueberries (Vaccinium SPP.) in Southern New Brunswick, Canada.</a> Biol. Conserv. 7, 301-309 (1975).</li><li>Crane, E., Walker, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The impact of pest management on bees and pollination. , (1983).</li><li>Haouar, M., Decormis, L., Rey, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluvalinate+applied+to+flowering+apple+trees-contamination+of+honey-gathering+bees+and+hive+products.">Fluvalinate applied to flowering apple trees-contamination of honey-gathering bees and hive products.</a> Agronomie. 10 (2), 133-137 (1990).</li><li>Chauzat, M. P., 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=A+survey+of+pesticide+residues+in+pollen+loads+collected+by+honey+bees+in+France.">A survey of pesticide residues in pollen loads collected by honey bees in France.</a> J. Econ. Entomol. 99 (2), 253-262 (2006).</li><li>Bonzini, S., Tremolada, P., Bernardinelli, I., Colombo, M., Vighi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Predicting+pesticide+fate+in+the+hive+(part+1),+experimentally+determined+&#964;-fluvalinate+residues+in+bees,+honey+and+wax.">Predicting pesticide fate in the hive (part 1), experimentally determined &#964;-fluvalinate residues in bees, honey and wax.</a> Apidologie. 42 (3), 378 (2011).</li><li>Sanchez-Bayo, F., Goka, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pesticide+residues+and+bees-a+risk+assessment.">Pesticide residues and bees-a risk assessment.</a> PLoS One. 9 (4), e94482 (2014).</li><li>Johnson, R. M., Ellis, M. D., Mullin, C. A., Frazier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pesticides+and+honey+bee+toxicity-USA.">Pesticides and honey bee toxicity-USA.</a> Apidologie. 41, 312-331 (2010).</li><li>Yang, E. C., Chang, H. C., Wu, W. Y., Chen, Y. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+olfactory+associative+behavior+of+honeybee+workers+due+to+contamination+of+imidacloprid+in+the+larval+stage.">Impaired olfactory associative behavior of honeybee workers due to contamination of imidacloprid in the larval stage.</a> PLoS One. 7, e49472 (2012).</li><li>Urlacher, E., 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=Measurements+of+chlorpyrifos+levels+in+forager+bees+and+comparison+with+levels+that+disrupt+honey+bee+odor-mediated+learning+under+laboratory+conditions.">Measurements of chlorpyrifos levels in forager bees and comparison with levels that disrupt honey bee odor-mediated learning under laboratory conditions.</a> J. Chem. Ecol. 42 (2), 127-138 (2016).</li><li>Chen, Y. W., Wu, P. S., Yang, E. C., Nai, Y. S., Huang, Z. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+pyriproxyfen+on+the+development+of+honey+bee+(Apis+mellifera+L.)+colony+in+field.">The impact of pyriproxyfen on the development of honey bee (Apis mellifera L.) colony in field.</a> J. Asia Pac. Entomol. 19 (3), 589-594 (2016).</li><li>Dennehy, T. J., DrGain, B. A., Harpold, V. S., Brink, S. A., Nichols, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Whitefly Resistance to Insecticides in Arizona: 2002 and 2003 Results. , 1926-1938 (2004).</li><li>Yang, E. C., Wu, P. S., Chang, H. C., Chen, Y. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+sub-lethal+dosages+of+insecticides+on+honey+bee+behavior+and+physiology.">Effect of sub-lethal dosages of insecticides on honey bee behavior and physiology.</a> , (2010).</li><li>Fourrier, J., 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=Larval+exposure+to+the+juvenile+hormone+analog+pyriproxyfen+disrupts+acceptance+of+and+social+behavior+performance+in+adult+honey+bees.">Larval exposure to the juvenile hormone analog pyriproxyfen disrupts acceptance of and social behavior performance in adult honey bees.</a> PLoS One. 10, e0132985 (2015).</li><li>Hanley, A. V., Huang, Z. Y., Pett, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+dietary+transgenic+Bt+corn+pollen+on+larvae+of+Apis+mellifera+and+Galleria+mellonella.">Effects of dietary transgenic Bt corn pollen on larvae of Apis mellifera and Galleria mellonella.</a> J. Apicult.Res. 42 (4), 77-81 (2003).</li><li>Kaftanoglu, O., Linksvayer, T. A., Page, 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=Rearing+honey+bees,+apis+mellifera,+in+vitro+1,+effects+of+sugar+concentrations+on+survival+and+development.">Rearing honey bees, apis mellifera, in vitro 1, effects of sugar concentrations on survival and development.</a> J. Insect Sci. 11 (96), 1-10 (2011).</li><li>Vandenberg, J. D., Shimanuki, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technique+for+rearing+worker+honey+bees+in+the+laboratory.">Technique for rearing worker honey bees in the laboratory.</a> J. Apicult. Res. 26 (2), 90-97 (1987).</li><li>Peng, Y. S. C., Mussen, E., Fong, A., Montague, M. A., Tyler, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+chlortetracycline+on+honey+bee+worker+larvae+reared+in+vitro.">Effects of chlortetracycline on honey bee worker larvae reared in vitro.</a> J. Invertebr.Pathol. 60 (2), 127-133 (1992).</li><li>Bitondi, M. M., Mora, I. M., Simoes, Z. L., Figueiredo, V. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Apis+mellifera+pupal+melanization+program+is+affected+by+treatment+with+a+juvenile+hormone+analogue.">The Apis mellifera pupal melanization program is affected by treatment with a juvenile hormone analogue.</a> J. Insect Physiol. 44 (5-6), 499-507 (1998).</li><li>Zufelato, M. S., Bitondi, M. M., Simoes, Z. L., Hartfelder, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+juvenile+hormone+analog+pyriproxyfen+affects+ecdysteroid-dependent+cuticle+melanization+and+shifts+the+pupal+ecdysteroid+peak+in+the+honey+bee+(Apis+mellifera).">The juvenile hormone analog pyriproxyfen affects ecdysteroid-dependent cuticle melanization and shifts the pupal ecdysteroid peak in the honey bee (Apis mellifera).</a> Arthropod Struct. Dev. 29 (2), 111-119 (2000).</li><li>Santos, A. E., Bitondi, M. M., Simoes, Z. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hormone-dependent+protein+patterns+in+integument+and+cuticular+pigmentation+in+Apis+mellifera+during+pharate+adult+development.">Hormone-dependent protein patterns in integument and cuticular pigmentation in Apis mellifera during pharate adult development.</a> J. Insect Physiol. 47 (11), 1275-1282 (2001).</li><li>Brouwers, E. V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose/Fructose+ratio+in+the+food+of+honeybee+larvae+during+caste+differentiation.">Glucose/Fructose ratio in the food of honeybee larvae during caste differentiation.</a> J. Apicult.Res. 23 (2), 94-101 (1984).</li><li>Howe, S. R., Dimick, P. S., Benton, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Composition+of+freshly+harvested+and+commercial+royal+jelly.">Composition of freshly harvested and commercial royal jelly.</a> J. Apicult. Res. 24 (1), 52-61 (1985).</li></ol>

저자는 더 경쟁 재정적 이해 관계가 없음을 선언합니다.

여왕 제한 알을 낳는 방법과 퀸 교환 방법은이 프로토콜 내에서 필드 테스트를 위해 꿀벌 그룹을 설정하기위한 중요한 단계이다. 여왕 제한 알을 낳는 방법은 꿀벌의 수명주기의 동기화를 가능하게한다. 그 결과, 연구진은 살충제의 서로 다른 용량으로 치료를 위해 같은 나이 1 일 된 유충을 선택할 수 있습니다. 퀸 교환 방법, 여왕이 부 A (4 개 프레임) 및 살충제 살충제 잔류 물의 영향을 평가하?...

환경에서 농약의 존재는 가장 심각한 문제 중 하나 인 영향을 미치는 꿀 꿀벌 1, 2, 3의 생명. 몇몇 연구는 꿀벌 식민지 꿀벌 제품에서 살충제 잔류의 일반적인 존재를 증명하고있다. 대만, 농약의 평균 응용 프로그램 11-12kg / (2005 년부터 2013 년) 매년 하였다. 대만에서 사용되는 농약의 양이 5 EU 국가보다 높은, 그리고 라틴 아메리카 국가 4. 즉, apicultural 환경은 특히 대만과 가능성이 다른 국가에서 심각한 농약 스트레스를 겪고있다. 꿀벌 API를 mellifera이 농업 시스템 (6)의 주요 매개자 중 하나이며 또한 꿀 가치있는 제품을 생산하고 있습니다. 그러나 꿀벌은 박람회는다양한 농약이 농약에 대한 ED 꿀과 꽃가루 7, 8을 수집 할 때 농약을 살포 한 꽃에서 먹이 찾아 돌아다 후 벌통에 다시 가져올 수 있습니다. 또한 자체가 두드러기 9, 10, 11 내 해충 문제를 제어하기 위해 목표로 양봉에 의해 농약에 노출 될 수 있습니다. 꿀벌 유충이 자신의 개발, 유충, 무인 항공기에 대한 간호사 꿀벌에 의해 공급되고 있기 때문에, 심지어 여왕은이 농약으로 오염 된 감로 및 꽃가루 (12)에 노출 될 수 있습니다. 꿀벌에 대한 다양한 농약의 독성 (13)를 해결해야합니다. 많은 노력은 환경 잔류 농약의 문제를 평가하기 위해 이루어졌다. 양 외. 에서 꿀벌 유충의 개발에 신경 독성 살충제 이미다클로 프리드의 영향을 테스트이 벌집과 이미다클로 프리드의 서브 치사량은 성인 꿀벌 후각 (14)의 결합 동작의 결과를보고했다. 또한 Urlacher 등. 실험실 조건 (15)에서 꿀 꿀벌 노동자의 학습 성능에 유기 인계 살충제, 클로르 피리 포스의 하위 치명적인 영향을 조사했다. 이전 연구에서 우리는 애벌레 꿀벌 16 (PPN) pyriproxyfen 곤충 생장 조절제의 영향 (IGR)을 평가. 본 논문에서는 꿀벌의 발전에 화학적 영향을 평가하기위한 방법을 제시한다. 꿀벌 공급 방법을 설명하고 하나의 개별 꿀벌 또는 콜로니에 적용 하였다. 첫째, 우리는 생체 내에서 개별 꿀벌에 대한 농약의 영향을 평가하기 위해 식민지에서 유충에 농약에 오염 된 기본 애벌레 다이어트 (BLD)의 서로 다른 농도를 시험했다. 우리는 자연 CONDIT을 시뮬레이션 진행벌통 내에 농약 오염 시럽을 사용하여 이온 살충제. 이 방법에서, 널리 해충 (17)에 대해 사용되며, 꿀벌 유충 및 번데기 (16), (18, 19)의 발전에 유해하다 PPN은 현장에서 농약의 부정적인 효과를 나타내는 지표가 될 것이다.

<table><tbody><tr><td>Honey bee box</td><td>SAN-YI Honey Factory</td><td>W1266</td><td>Honeybees rearing</td></tr><tr><td>Queen excluder (between frames)</td><td>SAN-YI Honey Factory</td><td>I1575</td><td>Queen limitation&amp;nbsp;</td></tr><tr><td>Queen excluder (on top)</td><td>SAN-YI Honey Factory</td><td>I1566</td><td>Queen limitation on top&amp;nbsp;</td></tr><tr><td>Bee brush</td><td>SAN-YI Honey Factory, Taiwan</td><td>W1414</td><td>clean the bees on frame gently</td></tr><tr><td>Bee feeder</td><td>SAN-YI Honey Factory, Taiwan</td><td>P0219</td><td>feed sugar syrup to colony</td></tr><tr><td>Transparent slide</td><td>Wan-Shih-Chei, Taiwan (http://www.mbsc.com.tw/a01goods.asp?s_id=40)</td><td>1139</td><td>Mark the larval area on the frames (Material: Polyethylene Terephthalate, PET) (Size = Length*Width*thick = 29.7 mm * 21 mm * 0.1 mm)</td></tr><tr><td>24 well tissu culture plate</td><td>Guangzhou Jet Bio-Filtration Co., Ltd</td><td>TCP011024</td><td>Rearing pupae from extraction</td></tr><tr><td>Autoclave</td><td>Tomin medical equipmenco., LTD.</td><td>TM-321</td><td>Make sterilized distilled deionized water (ddH&lt;sub&gt;2&lt;/sub&gt;O)</td></tr><tr><td>P20 pipetman</td><td>Gilson</td><td>F123600</td><td>Add PPN into bee larval food pool</td></tr><tr><td>Incubator&amp;nbsp;</td><td>Yihder Co., Ltd.</td><td>LE-550RD</td><td>Rearing pupae from extraction</td></tr><tr><td>Kimwipes</td><td>COW LUNG INSTRUMENT CO., LTD</td><td>KCS34155</td><td>Rearing pupae from extraction</td></tr><tr><td>Royal jelly</td><td>National Ilan University (NIU)</td><td>NIU</td><td>Make basic larval diet (BLD)</td></tr><tr><td>D-(+)-Glucose</td><td>Sigma</td><td>G8270</td><td>Make basic larval diet (BLD)</td></tr><tr><td>D-(-)-Fructose</td><td>Sigma</td><td>F0127</td><td>Make basic larval diet (BLD)</td></tr><tr><td>Yeast extract</td><td>CONDA, pronadisa</td><td>1702</td><td>Make basic larval diet (BLD)</td></tr><tr><td>Sucrose</td><td>Taiwan sugar coporation</td><td>E01071010</td><td>Make sugar syrup for bee food</td></tr><tr><td>Pyriproxyfen (11%)</td><td>LIH-NUNG CHEMICAL CO.. LTD.</td><td>Registration No. 1937</td><td>Insect growth regulator (IGR) used in the experiment</td></tr></tbody></table>

evaluating the effect of environmental chemicals on honey bee development from the individual to colony level

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be brought back to the beehive after the bees have foraged on flowers that have been sprayed with pesticides. Pesticide contaminated food can be exchanged between workers which then feed larvae and therefore can potentially affect the development of honey bees. Thus, residual pesticides in the environment can become a chronic damaging factor to honey bee populations and gradually lead to colony collapse. In the presented protocol, honey bee feeding methods are described and applied to either an individual honey bee or to a colony. Here, the insect growth regulator (IGR) pyriproxyfen (PPN), which is widely used to control pest insects and is harmful to the development of honey bee larvae and pupae, is used as the pesticide. The presenting procedure can be applied to other potentially harmful chemicals or honeybee pathogens for further studies.

꿀벌 필드 시험, 여왕이 산란을위한 4 프레임 부에 한정되었다. 이 단계는 한 프레임에 무리 밀도를 증가시키고 이후의 관찰을 용이하게 할 수있다. 각각의 치료는 표시되었고, 꿀벌 &#39;개발은 분명 투명 슬라이드를 통해 관찰되었다. 벌집 꿀벌 유충에 PPN-BLD의 생체 먹이 정확하게 콜로니에 꿀벌의 개발 PPN의 영향을 평가하기 위해 수행 하였다. 생체 공급 법?...

Watch this Scientific Journal Video about Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level at JoVE.com

Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

여기에서 우리는 제시 방법은 개별 꿀벌과 벌집 콜로니 모두 농약 오염 된 식품을 공급한다. 절차는 기본 유생 다이어트 생체 공급함으로써, 또한 벌집 콜로니의 자연 상태에서 개별 꿀벌의 살충제 효과를 평가한다.

식민지 수준에 개인에서 꿀 꿀벌 개발에 대한 환경 화학 물질의 효과를 평가

The presence of pesticides in the beekeeping environment is one of the most serious problems that impacts the life of a honey bee. Pesticides can be ...

evaluating-effect-environmental-chemicals-on-honey-bee-development

Universidad de Cartagena UDEC

Research

JoVE Journal

Environment

9.0K 조회수.  National Ilan University.  여기에서 우리는 제시 방법은 개별 꿀벌과 벌집 콜로니 모두 농약 오염 된 식품을 공급한다. 절차는 기본 유생 다이어트 생체 공급함으로써, 또한 벌집 콜로니의 자연 상태에서 개별 꿀벌의 살충제 효과를 평가한다. 

비디오: Evaluating the Effect of Environmental Chemicals on Honey Bee Development from the Individual to Colony Level

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA libraries are needed for optimal results from next-generation sequencing. Environmental samples such as water may have low quality and quantities of DNA as well as contaminants that co-precipitate with DNA. The mechanical and enzymatic processes involved in extraction and library preparation may further damage the DNA. Gel size selection enables purification and recovery of DNA fragments of a defined size for sequencing applications. Nevertheless, this task is one of the most time-consuming steps in the DNA library preparation workflow. The protocol described here enables complete automation of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments. 
In this study, we describe a high-throughput approach to prepare high quality DNA libraries from freshwater samples that can be applied also to other environmental samples. We used an indirect approach to concentrate bacterial cells from environmental freshwater samples; DNA was extracted using a commercially available DNA extraction kit, and DNA libraries were prepared using a commercial transposon-based protocol. DNA fragments of 500 to 800 bp were gel size selected using Ranger Technology, an automated electrophoresis workstation. Sequencing of the size-selected DNA libraries demonstrated significant improvements to read length and quality of the sequencing reads.

This manuscript describes an automated gel size selection approach for purifying DNA fragments for next-generation sequencing. The Ranger Technology provides complete automation of the entire process of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments allowing for high-throughput and high quality next-generation sequencing libraries.

자동 젤 사이즈 선택은 환경 물 샘플에서 준비 차세대 시퀀싱 라이브러리의 질을 향상시키기 위해

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA ...

연구

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 ...

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide regulation studies. The risk of exposure to fungicide residues is likely to be especially high if the spray occurs on, or near host plants while the bees are collecting pollen to provision their nests. For species of Osmia that consume pollen from a select group of plants (oligolecty), the inability to use pollen from non-host plants can increase their risk factor for fungicide-related toxicity. This manuscript describes protocols used to successfully rear oligolectic mason bees, Osmia ribifloris sensu lato, from egg to prepupal stage within cell culture plates under standardized laboratory conditions. The in vitro-reared bees are subsequently used to investigate the effects of fungicide exposure and pollen source on bee fitness. Based on a 2 &#215; 2 fully crossed factorial design, the experiment examines the main and interactive effects of fungicide exposure and pollen source on larval fitness, quantified by prepupal biomass, larval developmental time, and survivorship. A major advantage of this technique is that using in vitro-reared bees reduces natural background variability and allows the simultaneous manipulation of multiple experimental parameters. The described protocol presents a versatile tool for hypotheses testing involving the suite of factors affecting bee health. For conservation efforts to be met with significant, lasting success, such insights into the complex interplay of physiological and environmental factors driving bee declines will prove to be critical.

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors

Fungicide sprays on flowering plants may expose solitary bees to high concentrations of pollen-borne fungicide residues. Using laboratory-based experiments involving in vitro-reared bee larvae, this study investigates the interactive effects of consuming fungicide-treated pollen derived from host and non-host plants.

생체 외에서 고독한 꿀벌의 양육: 애벌레 위험 요소를 평가 하기 위한 도구

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide ...

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)

This video demonstrates novel techniques of RNA interference (RNAi) which downregulate two genes simultaneously in honey bees using double-stranded RNA (dsRNA) injections. It also presents a protocol of proboscis extension response (PER) assay for measuring gustatory perception.	
	RNAi-mediated gene knockdown is an effective technique downregulating target gene expression. This technique is usually used for single gene manipulation, but it has limitations to detect interactions and joint effects between genes. In the first part of this video, we present two strategies to simultaneously knock down two genes (called double gene knockdown). We show both strategies are able to effectively suppress two genes, vitellogenin (vg) and ultraspiracle (usp), which are in a regulatory feedback loop. This double gene knockdown approach can be used to dissect interrelationships between genes and can be readily applied in different insect species.	
	The second part of this video is a demonstration of proboscis extension response (PER) assay in honey bees after the treatment of double gene knockdown. The PER assay is a standard test for measuring gustatory perception in honey bees, which is a key predictor for how fast a honey bee's behavioral maturation is. Greater gustatory perception of nest bees indicates increased behavioral development which is often associated with an earlier age at onset of foraging and foraging specialization in pollen. In addition, PER assay can be applied to identify metabolic states of satiation or hunger in honey bees. Finally, PER assay combined with pairing different odor stimuli for conditioning the bees is also widely used for learning and memory studies in honey bees.

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees Apis mellifera

In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.

꿀벌의 RNAi를 매개로 두 유전자 넉다운과 미각 지각 측정 (<em&gt; 아피스 mellifera</em&gt;)

This video demonstrates novel techniques of RNA  interference (RNAi) which downregulate two genes simultaneously in honey bees  using double-stranded RNA ...

신경과학

Evaluating the Effect of Pesticides on the Larvae of the Solitary Bees

Current ecological risk assessments of pesticides on pollinators have primarily considered only laboratory conditions. For the larvae of solitary bees, ingestion of provisions contaminated with pesticides may increase the mortality rate of the larvae, decrease the collection rate and the population of adult solitary bees in the next year from a demographic perspective. But there are limited studies on the effects of pesticides on the larvae of solitary bees. Therefore, understanding how pesticides influence the larvae of solitary bees should be considered an integral part of pesticide ecological risk assessment. This study presents a method to expose the larvae of solitary bee, Osmia excavata, to lethal or sublethal doses of pesticide, tracking larval weight gain, developmental duration, eclosion ability, and food consumption efficiency conversion of ingested food. To demonstrate the effectiveness of this method, the larvae of O. excavata were fed with provisions containing acute lethal and sublethal doses of chlorpyrifos. Then, the above indexes of the treated larvae were investigated. This technique helps to predict and mitigate the risk of pesticides to pollinators.

The present protocol explains a method to feed pesticide-contaminated provisions to the larvae of the solitary bees, Osmia excavata. The procedure examines the ecotoxicity of the pesticide to the larvae of the solitary bees.

살충제가 독방 꿀벌의 유충에 미치는 영향 평가

Current ecological risk assessments of pesticides on pollinators have primarily considered only laboratory conditions. For the larvae of solitary bees, ...

생화학

Preparation of Single-cohort Colonies and Hormone Treatment of Worker Honeybees to Analyze Physiology Associated with Role and/or Endocrine System

Honeybee workers are engaged in various tasks related to maintaining colony activity. The tasks of the workers change according to their age (age-related division of labor). Young workers are engaged in nursing the brood (nurse bees), while older workers are engaged in foraging for nectar and pollen (foragers). The physiology of the workers changes in association with this role shift. For example, the main function of the hypopharyngeal glands (HPGs) changes from the secretion of major royal jelly proteins (MRJPs) to the secretion of carbohydrate-metabolizing enzymes. Because worker tasks change as the workers age in typical colonies, it is difficult to discriminate the physiological changes that occur with aging from those that occur with the role shift. To study the physiological changes in worker tissues, including the HPGs, in association with the role shift, it would be useful to manipulate the honeybee colony population by preparing single-cohort colonies in which workers of almost the same age perform different tasks. Here we describe a detailed protocol for preparing single-cohort colonies for this analysis. Six to eight days after single-cohort colony preparation, precocious foragers that perform foraging tasks earlier than usual appear in the colony. Representative results indicated role-associated changes in HPG gene expression, suggesting role-associated HPG function. In addition to manipulating the colony population, analysis of the endocrine system is important for investigating role-associated physiology. Here, we also describe a detailed protocol for treating workers with 20-hydroxyecdysone (20E), an active form of ecdysone, and methoprene, a juvenile hormone analogue. The survival rate of treated bees was sufficient to examine gene expression in the HPGs. Gene expression changes were observed in response to 20E- and/or methoprene-treatment, suggesting that hormone treatments induce physiological changes of the HPGs. The protocol for hormone treatment described here is appropriate for examining hormonal effects on worker physiology.

Here we describe our detailed protocol for the preparation of single-cohort honeybee colonies&#160;&#8211; a useful tool for analyzing the role-associated worker physiology. We also describe detailed protocols for treating workers with juvenile hormone and ecdysone to evaluate the involvement of these hormones in the regulation of worker behavior and/or physiology.

단일 코호트 식민지와 노동자 꿀벌의 호르몬 치료의 준비 역할 및 / 또는 내분비 시스템과 생리학 관련 분석하기

Honeybee workers are engaged in various tasks related to maintaining colony activity. The tasks of the workers change according to their age (age-related ...

SwarmSight: Real-time Tracking of Insect Antenna Movements and Proboscis Extension Reflex Using a Common Preparation and Conventional Hardware

Many scientifically and agriculturally important insects use antennae to detect the presence of volatile chemical compounds and extend their proboscis during feeding. The ability to rapidly obtain high-resolution measurements of natural antenna and proboscis movements and assess how they change in response to chemical, developmental, and genetic manipulations can aid the understanding of insect behavior. By extending our previous work on assessing aggregate insect swarm or animal group movements from natural and laboratory videos using the video analysis software SwarmSight, we developed a novel, free, and open-source software module, SwarmSight Appendage Tracking (SwarmSight.org) for frame-by-frame tracking of insect antenna and proboscis positions from conventional web camera videos using conventional computers. The software processes frames about 120 times faster than humans, performs at better than human accuracy, and, using 30 frames per second (fps) videos, can capture antennal dynamics up to 15 Hz. The software was used to track the antennal response of honey bees to two odors and found significant mean antennal retractions away from the odor source about 1 s after odor presentation. We observed antenna position density heat map cluster formation and cluster and mean angle dependence on odor concentration.

This protocol describes steps for using the novel software, SwarmSight, for frame-by-frame tracking of insect antenna and proboscis positions from conventional web camera videos using conventional computers. The free, open-source software processes frames about 120 times faster than humans and performs at better than human accuracy.

SwarmSight: 곤충 안테나와 일반적인 준비와 기존 하드웨어를 사용 하 여 코 확장 반사의 실시간 추적

Many scientifically and agriculturally important insects use antennae to detect the presence of volatile chemical compounds and extend their proboscis ...

Monitoring Colony-level Effects of Sublethal Pesticide Exposure on Honey Bees

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment methods. Here we describe methods to measure the quantities of adult bees, brood, and food resources by weighing hives and hive parts, by photographing frames, and by installing hives on scales and with internal sensors. Data from these periodic evaluations are then combined with running average and daily detrended data on hive weight and internal hive temperature. The resulting datasets have been used to detect colony-level effects of imidacloprid applied in a sugar syrup as low as 5 parts per billion. The methods are objective, require little training, and provide permanent records in the form of sensor output and photographs.

Sublethal doses of pesticides may affect colonies in ways that are difficult to detect using only periodic or visual methods. Methods to measure total adult bee mass, brood, and food resources by weighing hives and hive parts, photographing frames, and installing sensors, are provided. Data analysis is also addressed.

꿀 꿀벌에 Sublethal 농약 노출의 식민지 수준 효과 모니터링

The effects of sublethal pesticide exposure to honey bee colonies may be significant but difficult to detect in the field using standard visual assessment ...

생물학

Assessing Agrochemical Risk to Mated Honey Bee Queens

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not accurately capture the effects of agrochemical exposure on honey bee queens. As the sole producer of fertilized eggs inside a honeybee colony, the queen is arguably the most important single member of a functioning colony unit. Therefore, understanding how agrochemicals affect queen health and productivity should be considered a critical aspect of pesticide risk assessment. Here, an adapted method is presented to expose honey bee queens and worker queen attendants to agrochemical stressors administered through a worker diet, followed by tracking egg production in the laboratory and assessing first instar eclosion using a specialized cage, referred to as a Queen Monitoring Cage. To illustrate the method&#39;s intended use, results of an experiment in which worker queen attendants were fed diet containing sublethal doses of imidacloprid and effects on queens were monitored are described.

This protocol was developed to enhance the understanding of how agrochemicals affect honey bee (Apis mellifera) reproduction by establishing methods to expose honey bee queens and their worker caretakers to agrochemicals in a controlled, laboratory setting and carefully monitoring their relevant responses.

꿀벌 여왕을 짝짓기하는 농약적 위험 평가

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not ...

Histology Basics and Cell Death Detection in Honeybee Tissue

Honeybees (Apis mellifera L.) inside the hive (nurse workers and other hive bees) and outside the hive (foragers) are exposed to climate and weather changes, various pesticides, pathogens, and malnutrition, mainly entering through the mouth and primarily affecting the digestive tracts of adult bees. To understand and prevent the effects of such external and internal stressors on honeybees, one useful research method is the immunohistochemical method. A basic protocol is described to prepare the midgut (ventriculus) and hypopharyngeal glands (HPGs) of adult bees for histological analysis. A detailed methodology is described to assess the level of cell damage and distinguish necrosis from programmed cell death (apoptosis) as a natural process of tissue regeneration. The results of adult honeybee treatment with oxalic acid and pesticides (insecticide and acaricide) and the determination of cell death in the ventriculus and HPGs are presented. The pros and cons of the methodology are also discussed.

Immunohistochemical methods are useful in honeybee research to detect and assess the level of apoptosis and necrosis in the midgut and hypopharyngeal glands of adult bees.

꿀벌 조직의 조직학 기초 및 세포 사멸 검출

Honeybees (Apis mellifera L.) inside the hive (nurse workers and other hive bees) and outside the hive (foragers) are exposed to climate and weather ...