Questa ricerca è stata sostenuta da Grant 105AS-13.2.3-BQ-B1 dal Bureau of animali e piante di ispezione sanitaria e la quarantena, il Consiglio per l&#39;Agricoltura, Executive Yuan e Grant 103-2313-B-197-002-MY3 dal Ministero della Scienza e della Tecnologia (MOST).

1. preparativi <ol><li> Fare 1 L di 50% sciroppo di zucchero. Sciogliere 1 kg saccarosio in 1 L DDH 2 O. </li><li> Preparare la soluzione piriprossifen (PPN) in BLD. Rendere 1,1 L di 10.000 ppm PPN soluzione madre e diluire 100 ml di soluzione PPN in 1 L sterilizzato DDH 2 O. Conservare a 4 ° C. </li><li> Diluire la soluzione madre PPN a concentrazioni finali di 0,1, 1, 10 e 100 mg / kg (ppm) nella BLD per il seguente esperimento. </li><li> Fare PPN-sciroppo (per il livello di colonia). Diluire il brodo PPN a concentrazioni finali di 10 e 100 ppm nel 50% sciroppo di zucchero per il seguente esperimento. </li><li> Il miele delle api allevamento. Nota: Qui, la posizione sperimentale è l&#39;apiario nazionale Ilan University (NIU), Yi-Lan City, Taiwan; (Coordinate GPS: N24.747278, E121.746200). <ol><li> Controllare miele ape (Apis mellifera L.) colonie settimanale per quantità di alimenti e mangimi con 1 litro di sciroppo di zucchero 50%, se necessario (l&#39;area di memorizzazione miele è vuoto). Definire un sanocolonia 9 fotogrammi di pettini api in ogni colonia con una regina che le uova normalmente. </li></ol></li><li> Preparare 100 ml di dieta larvale di base (BLD). Sciogliere 6% D-glucosio, fruttosio 6% e 1% di estratto di lievito in acqua deionizzata distillata sterilizzata (DDH 2 O) e integra con il 50% gelatina reale. Conservare a 4 ° C, ma non per più di 3 giorni. Nota: Pre-caldo a 35 ° C prima dell&#39;esperimento e consumare entro 3 giorni. </li></ol> 2. In Vivo metodo di alimentazione Nota: In vivo metodo di alimentazione è stato modificato da Hanley et al. 20 <ol><li> Il miele delle api selezione larve e l&#39;etichettatura. <ol><li> Inserire un escludi regina dividere 9 fotogrammi di una colonia sana in 4 frame e 5 fotogrammi limitando la regina alla sezione 4 telaio. Lasciare almeno un fotogramma vuoto nella sezione 4 telaio per la posa delle uova. </li><li> Dopo la regina deporre le uova per 1 giorno, Controllare i telai per la comparsa di uova e mantenere le uova all&#39;interno delle arnie per 72 ore (il 4 ° giorno) fino al 1 ° giorno di vita schiusa delle larve. Prendete una delle cornici contenenti 1-giorni di età le larve dei lavoratori (tratteggiata entro 24 ore) fuori dall&#39;alveare di prova a mano e togliere i lavoratori api dal telaio con una spazzola ape. </li><li> Coprire il telaio con un vetrino trasparente (Formato = lunghezza x larghezza x spessore = 29,7 millimetri x 21 mm x 0,1 mm) e inchiodare il vetrino trasparente sul bordo dei telai con puntine (Figura 1A). </li><li> Per ogni trattamento, selezionate casualmente 50 una larva di un giorno (il 4 ° giorno) e contrassegnare ciascuna cella covata utilizzando pennarelli permanenti sul vetrino trasparente (Figure 1A e 1B). Nota: scrivere l&#39;informazione di ogni trattamento sul telaio e scorrevole trasparente nonché utilizzando pennarelli permanenti per evitare la confusione tra diversi trattamenti. Rimuovere i vetrini segnalati e conservare per l&#39;in vivo &lt;/ Em&gt; alimentazione e osservazione. </li></ol></li><li> Nell&#39;alimentazione vivo. <ol><li> Aggiungere diverse concentrazioni di PPN-BLD (0,1, 1, 10 e 100 ppm) a ciascuna cella covata marcato pipettando al giorno 1, 2 e 3 con 10 microlitri, 10 microlitri e 20 uL, rispettivamente, secondo la quantità di aspirazione del età larve. Aggiungere la stessa quantità di BLD (senza PPN) ad un gruppo di controllo. In tal modo, la dose totale di PPN-BLD in ogni cella covata etichettati accumula 4, 40, 400, e 4000 pg. NOTA: Riconoscere le celle di covata etichettati dai vetrini trasparenti segnalati e rimuovere i vetrini trasparenti selezionati dopo l&#39;alimentazione. Utilizzare fresco BLD per l&#39;alimentazione per prevenire la pulizia delle celle di covata dalle api operaie. </li><li> Riportare i telai PPN-trattati nelle colonie originali per ulteriori osservazioni. NOTA: Ogni trattamento ha quattro ripete biologici in quattro colonie. </li></ol></li><li> L&#39;osservazione delle larve trattate al giorno 7. <ol><li> Per osservare °larve e PPN-trattati, restituire le diapositive trasparenti etichettati ai telai per registrare la mortalità e tassi di tappatura nelle celle di covata etichettati. </li></ol></li><li> L&#39;osservazione dei tassi di pupe e Eclosion trattati a giorno 13. <ol><li> Rimuovere la cera d&#39;api di celle di covata innevate. </li><li> Mettere le pinzette morbida capovolti nella cella di covata e serrare le pupe leggermente poi prendere pupe con delicatezza. </li><li> Trasferire il pupe in piastre da 24 pozzetti di coltura tissutale con un doppio strato di tessuto di laboratorio sotto ogni pozzetto. Registrare il danno e la mortalità durante il trasferimento di pupa. </li><li> Mantenere le piastre da 24 pozzetti di coltura tissutale in un incubatore a 34 ° C e 70% di umidità relativa fino emergere (circa 8 giorni). </li><li> Osservare e registrare le pupe e api mellifere emerse. </li></ol></li><li> statistica <ol><li> Calcolare i dati registrati e presente come media ± SD </li><li> Analizzare i dati utilizzando l&#39;analisi della varianza (ANOVA) da SAS e utilizzare il test minima differenza significativa (LSD) per analizzare le differenze tra due mezzi diversi trattamenti. Definire statisticamente significativo come P -value &lt;0.05. Lettere diverse nella stessa colonna della tabella mostrato un effetto significativo dall&#39;analisi statistica. </li></ol></li></ol> 3. Tossicità dei PPN a Colony Livello nella Beehive <ol><li> Impostare i gruppi di api. <ol><li> Inserire un escludi regina verticalmente dividere 9 fotogrammi di una colonia sana in 4 fotogrammi (parte A) e 5 fotogrammi (parte B), limitando la regina alla sezione 4 telaio. Lasciare almeno un fotogramma vuoto nella sezione 4-struttura per la deposizione delle uova. Nota: In altre escludi-regina del sulla parte superiore della parte della regina per evitare che la regina di muoversi tra le parti. </li><li> Dopo la regina deporre le uova per 1 giorno, controllare i telai per la comparsa di uova. Prendere i fotogrammi appropriati contenenti uova fuori dalla parte A a mano e togliere i lavoratori apidal telaio con una spazzola ape. </li><li> Coprire il telaio con scivolo trasparente e inchiodare il vetrino trasparente al bordo del telaio con puntine. </li><li> Selezionare casualmente 100 celle di covata contenenti uova e contrassegnare ciascuna cella covata utilizzando pennarello indelebile sul vetrino trasparente. Assegnare queste celle di covata 100 come gruppo 1. scrivere le informazioni di ciascun trattamento sul telaio e scorrevole trasparente con pennarello indelebile per evitare la confusione tra diversi trattamenti. </li><li> Riportare la cornice etichettati per parte A per 3 giorni e poi trasferire la regina parte B per deporre le uova. </li><li> Dopo 1 giorno, controllare i fotogrammi di parte B, scegliere una cornice adeguata ed etichetta 100 celle di covata contenenti uova come descritto al punto 3.1.4. Assegnare queste celle di covata 100 come gruppo 2. </li><li> Riportare la cornice etichettati per parte B per 3 giorni e poi trasferire la regina alla parte A per deporre le uova. </li><li> Ripetere lo scambio regina tra le parti A e B 6 volte di più e assegnare gruppi numericamente,rispettivamente (Figura 2). Ci dovrebbe essere totale di 9 gruppi. </li></ol></li><li> T miele reat colonia di api con PPN sciroppo di zucchero al giorno 13 (Figura 2). <ol><li> Aggiungere 1 L 50% PPN sciroppo di zucchero contenenti 10 o 100 ppm in una cassa di alimentazione ape plastica (L x L x H = 20 cm x 30 cm x 3,5 cm) e mettere la scatola in cima delle cornici nelle colonie sperimentali. Nota: Gruppo 1 non riceve il PPN a 13 giorni, come le celle di covata sono stati sigillati. </li><li> Alimentare il gruppo di controllo con 1 L 50% di sciroppo di zucchero (senza PPN) come descritto al punto 2.2. </li></ol></li><li> Conta 1 giorno di età larve e record come il tasso di schiusa dell&#39;uovo di 100 cellule marcate covata per ciascun gruppo a giorno 5 (Figura 2). Miele api uova di solito prendere 3 giorni per schiudersi in 0 giorni larve; quindi, controllare e etichettare 100 uova-contenenti celle di covata al giorno 1 e contare il numero di 1 giorno di età larve per ogni gruppo di giorno 5 per ottenere la percentuale di schiusa tasso. </li><li> CMONTAGGIO DEL celle di covata ricoperti e registrare come il tasso massimo larvale di 100 cellule marcate covata per ciascun gruppo a giorno 11 (Figura 2). 6 a 7 celle di covata larvale di un giorno saranno ricoperti con cera d&#39;api da parte dei lavoratori ape per le larve pupating. </li><li> Osservare pupe maturazione e registrare il tasso sfarfallamento di 100 cellule marcate covata per ciascun gruppo a giorno 17 (Figura 2). <ol><li> Rimuovere la cera d&#39;api di celle di covata capped e prendere pupe con morbidi pinzetta a punta delicatamente. Trasferire il pupe in un piastre di coltura tissutale a 24 pozzetti con un doppio strato di tessuto di laboratorio sotto ogni pozzetto. </li><li> Mantenere le piastre da 24 pozzetti di coltura tissutale in un incubatore a 34 ° C e 70% di umidità relativa fino emersione. </li><li> Osservare e registrare le pupe ed emerse le api per ogni gruppo fino Gruppo 9 (49 giorni sperimentali). Nota: Ogni trattamento ha quattro ripete biologici. </li></ol></li><li> statistica <ol><li> Calcolare i dati registrati e presenti come mediaSD ±. </li><li> Analizzare le differenze significative tra coppie di trattamenti (per esempio 0 ppm / 10 ppm, 10 ppm / 100 ppm e 0 ppm / 100 ppm) in ogni gruppo utilizzando t-test a due code di Student. Definire come statisticamente significativo se P -value &lt;0.05. </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>

Gli autori dichiarano di non avere interessi finanziari concorrenti.

Il metodo di deposizione delle uova queen-limitata e il metodo queen-scambio sono passaggi critici per la creazione di gruppi di api per le prove sul campo all&#39;interno di questo protocollo. Il metodo regina limitata ovodeposizione permette la sincronizzazione del ciclo di vita delle api. Di conseguenza, i ricercatori possono selezionare 1 giorno-vecchio larve della stessa età per il trattamento con diverse dosi di pesticidi. Per il metodo francese, lo scambio, la regina è stato scambiato tra la parte A (4 fotogram...

La presenza di pesticidi nell&#39;ambiente è uno dei problemi più gravi che l&#39;impatto della vita di un&#39;ape 1, 2, 3. Diversi studi hanno dimostrato la presenza di residui di pesticidi comuni in colonie di api mellifere e prodotti delle api. A Taiwan, l&#39;applicazione di pesticidi media era 11-12 kg / ha ogni anno (2005-2013). La quantità di pesticidi utilizzati a Taiwan è superiore a quella dei paesi dell&#39;UE, ei paesi dell&#39;America Latina 4, 5. In altre parole, l&#39;ambiente all&#39;apicoltura sta soffrendo gravi sollecitazioni di pesticidi, soprattutto a Taiwan e forse anche in altri paesi. Il miele ape mellifera è uno dei principali impollinatori nei sistemi agricoli 6 e produce anche prodotti pregiati come miele. Tuttavia, le api sono manifestazioniEd a vari pesticidi e questi pesticidi può essere riportato al alveari dopo foraggiamento sui fiori che sono stati spruzzati con pesticidi quando si raccolgono nettare e polline 7, 8. Essi inoltre possono essere esposti ai pesticidi dagli apicoltori stessi che mirano a controllare i problemi dei parassiti all&#39;interno delle arnie 9, 10, 11. Perché il miele delle api larve sono alimentati da api nutrici per il loro sviluppo, le larve, droni e perfino la regina può essere esposto a questi nettari pesticidi contaminati e polline 12. La tossicità dei vari pesticidi per le api deve essere affrontato 13. Molti sforzi sono stati fatti per valutare le emissioni di residui di pesticidi ambientali. Yang et al. testato l&#39;influenza del imidacloprid insetticidi neurotossici sullo sviluppo di miele delle api larve nelalveare e ha riferito che una dose sub-letali di imidacloprid ha provocato un comportamento associativo olfattiva delle api adulte 14. Inoltre, Urlacher et al. ha esaminato gli effetti sub-letali di una pesticidi organofosfati, clorpirifos, sulle prestazioni di apprendimento un miele di ape operaia in condizioni di laboratorio 15. Nel nostro studio precedente, abbiamo valutato l&#39;impatto di un regolatore di crescita degli insetti (IGR), piriprossifen (PPN), su api da miele larvali 16. In questo documento, presentiamo metodi per valutare gli effetti chimici sullo sviluppo delle api. Miele metodi di alimentazione ape sono stati descritti ed applicati sia singole api mellifere o ad una colonia. In un primo momento, abbiamo provato diverse concentrazioni di pesticidi contaminato dieta larvale di base (BLD) di larve nelle colonie per valutare l&#39;impatto del pesticida sulle singole api mellifere in vivo. Si è quindi proceduto a simulare la Condit naturaliioni del pesticida utilizzando lo sciroppo di pesticidi contaminati all&#39;interno di alveari. In questo metodo, PPN, che è ampiamente usato contro insetti parassiti 17 ed è dannoso per lo sviluppo di miele api larve e pupe 16, 18, 19, sarà un indicatore per rappresentare l&#39;effetto negativo del pesticida nel campo.

<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.

Per la prova sul campo miele ape, una regina era limitato alla sezione 4-struttura per la deposizione delle uova. Questa fase potrebbe aumentare la densità covata in un frame e facilitare osservazioni successive. Ogni trattamento è stato etichettato, e lo sviluppo delle api stato chiaramente osservato attraverso un vetrino trasparente. In vivo alimentazione di PPN-BLD al miele api larve dell&#39;alveare è stata eseguita per valutare con precisione l&#39;influenza della PPN su...

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

Qui vi presentiamo un metodo per alimentare antiparassitario alimenti contaminati sia un individuo ape e una colonia alveare. La procedura valuta l&#39;effetto dei pesticidi sulle singole api mellifere in vivo l&#39;alimentazione della dieta larvale di base e anche la condizione naturale della colonia dell&#39;alveare.

Valutare l&#39;effetto delle sostanze chimiche ambientali su Honey Bee sviluppo dall&#39;individuo alla colonia Livello

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 Views.  National Ilan University.  Qui vi presentiamo un metodo per alimentare antiparassitario alimenti contaminati sia un individuo ape e una colonia alveare. La procedura valuta l'effetto dei pesticidi sulle singole api mellifere in vivo l'alimentazione della dieta larvale di base e anche la condizione naturale della colonia dell'alveare. 

Video: 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.

Automated Gel Selection Size per migliorare la qualità delle Biblioteche di sequenziamento di nuova generazione Preparato da campioni di acqua ambientale

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

Ricerca

Ambiente

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.

Empirica, metagenomica e tecniche computazionali illuminano i meccanismi attraverso i quali salute delle API di fungicidi compromesso

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.

In Vitro Allevamento di api solitarie: uno strumento per valutare i fattori di rischio larvale

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.

RNAi mediato doppia Gene atterramento e gustative Percezione misura di api mellifere (<em&gt; Apis mellifera</em&gt;)

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

Neuroscienze

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.

Valutare l'effetto dei pesticidi sulle larve delle api solitarie

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

Biochimica

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.

Preparazione di singole colonie-coorte e trattamento ormonale di lavoratore api per analizzare Fisiologia associati con il ruolo e / o sistema endocrino

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: Monitoraggio in tempo reale dei movimenti di Antenna dell'insetto e proboscide estensione Reflex utilizzando una preparazione comune e Hardware convenzionali

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.

Monitoraggio effetti Colonia-livello di esposizione dell'antiparassitario subletali sulle API di miele

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

Biologia

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.

Valutazione del rischio agrochimico per le regine delle api da miele accoppiate

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.

Nozioni di base sull'istologia e rilevamento della morte cellulare nel tessuto delle api

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

Valutare l'effetto delle sostanze chimiche ambientali su Honey Bee sviluppo dall'individuo alla colonia Livello

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