Name: evaluating the effect of environmental chemicals on honey bee development from the individual to colony level
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This research was supported by Grant 105AS-13.2.3-BQ-B1 from the Bureau of Animal and Plant Health Inspection and Quarantine, the Council of Agriculture, Executive Yuan and Grant 103-2313-B-197-002-MY3 from the Ministry of Science and Technology (MOST).

1. Voorbereidingen <ol><li> Maak 1 liter 50% suikerstroop. Los 1 kg sucrose in 1 L DDH 2 O. </li><li> Bereid pyriproxyfen (PPN) oplossing in BLD. Voeg 1,1 l 10.000 ppm PPN voorraadoplossing en verdun 100 ml PPN-oplossing in 1 liter gesteriliseerde DDH 2 O. Bewaar bij 4 ° C. </li><li> Verdun de PPN voorraadoplossing tot eindconcentraties van 0,1, 1, 10 en 100 mg / kg (ppm) in de BLD voor het volgende experiment. </li><li> Maak PPN-siroop (voor de kolonie niveau). Verdun het PPN stock eindconcentr...

<ol>
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The queen-limited egg-laying method and queen-exchange method are critical steps for setting up honey bee groups for field testing within this protocol. The queen-limited egg-laying method permits synchronization of the life cycle of honey bees. Consequently, researchers can select 1 day-old larvae of the same age for treatment with different doses of pesticide. For the queen-exchange method, the queen was exchanged between part A (4 frames) and B (5 frames) to obtain different developmental stages of honey bee for field...

De aanwezigheid van pesticiden in het milieu is een van de meest ernstige problemen die invloed heeft op de levensduur van een honingbij 1, 2, 3. Verschillende studies hebben de gemeenschappelijke aanwezigheid van residuen van bestrijdingsmiddelen in bijenkolonies en bijenproducten aangetoond. In Taiwan, het gemiddelde gebruik van pesticiden was 11-12 kg / ha per jaar (2005-2013). De hoeveelheid gebruikte pesticiden in Taiwan is hoger dan die van de EU-landen en de Latijns-Amerikaanse landen 4, 5. Met andere woorden, de bijenteelt milieu ernstige pesticide stress, vooral in Taiwan en mogelijk ook in andere landen. De honingbij Apis mellifera is één van de belangrijkste bestuivers in landbouwsystemen 6 en het produceert ook waardevolle producten zoals honing. Echter, honingbijen zijn expoed aan verschillende pesticiden en deze bestrijdingsmiddelen kunnen terug naar bijenkorven na het foerageren op de bloemen die zijn bespoten met bestrijdingsmiddelen bij het verzamelen van nectar en stuifmeel 7, 8 worden gebracht. Ze kunnen ook worden blootgesteld aan pesticiden door de bijenhouders als doel plagen in de bijenkast 9, 10, 11 te regelen. Omdat honingbij larven worden gevoed door verpleegkundige bijen voor hun ontwikkeling, larven, drones en zelfs de koningin kan worden blootgesteld aan deze pesticide verontreinigde nectar en stuifmeel 12. De toxiciteit van verschillende bestrijdingsmiddelen honingbijen moet worden aangepakt 13. Vele pogingen zijn gedaan om de problemen van milieu-residuen van bestrijdingsmiddelen te evalueren. Yang et al. testte de invloed van de neurotoxische insecticide imidacloprid op de ontwikkeling van de honingbij larven in debijenkorf en gemeld dat een sub-dodelijke dosis imidacloprid resulteerde in olfactorische associatief gedrag van de volwassen bijen 14. Ook Urlacher et al. onderzochten de sub-letale effecten van een organofosfaat pesticide, chloorpyrifos, op leerprestaties een honingbij werknemer onder laboratoriumomstandigheden 15. In onze vorige studie, evalueerden we de impact van een insect groei regulator (IGR), pyriproxyfen (PPN), op larvale honingbijen 16. In deze paper presenteren we methoden voor de evaluatie van de chemische effecten op de ontwikkeling van de honingbijen. Honingbij voedermethoden werden beschreven en aangebracht op ofwel individuele bijen of een kolonie. In eerste instantie hebben we getest verschillende concentraties van pesticiden verontreinigde basic larvale dieet (BLD) op larven in de koloniën om de impact van het pesticide op de individuele honingbijen in vivo te evalueren. We gingen vervolgens over tot de natuurlijke condit simulerenionen van het pesticide met pesticide verontreinigde siroop in bijenkorven. Bij deze werkwijze wordt PPN, die wijd tegen plaaginsecten 17 en is schadelijk voor de ontwikkeling van honingbij larven en poppen 16, 18, 19, een indicator om het negatieve effect van het pesticide op het gebied vertegenwoordigen.

<table border="0" cellpadding="0" cellspacing="0" width="887">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:193px;">Honey bee box</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">W1266</td>
			<td style="width:297px;">Honeybees rearing</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Queen excluder (between frames)</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">I1575</td>
			<td style="width:297px;">Queen limitation&#160;</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Queen excluder (on top )</td>
			<td style="width:243px;">SAN-YI Honey Factory</td>
			<td style="width:155px;">I1566</td>
			<td style="width:297px;">Queen limitation on top&#160;</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Bee brush</td>
			<td style="width:243px;">SAN-YI Honey Factory, Taiwan</td>
			<td>W1414</td>
			<td style="width:297px;">clean the bees on frame gently</td>
		</tr>
		<tr height="18">
			<td height="18" style="height:18px;width:193px;">Bee feeder</td>
			<td style="width:243px;">SAN-YI Honey Factory, Taiwan</td>
			<td>P0219</td>
			<td style="width:297px;">feed sugar syrup to colony</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;">Transparent slide</td>
			<td style="width:243px;">Wan-Shih-Chei, Taiwan (http://www.mbsc.com.tw/a01goods.asp?s_id=40)</td>
			<td>1139</td>
			<td style="width:297px;">Mark the larval area on the frames (Material: Polyethylene Terephthalate, PET) (Size= Length*Width*thick= 29.7mm*21mm*0.1mm)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">24 well tissu culture plate</td>
			<td style="width:243px;">Guangzhou Jet Bio-Filtration Co., Ltd</td>
			<td style="width:155px;">TCP011024</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Autoclave</td>
			<td style="width:243px;">Tomin medical equipmenco., LTD.</td>
			<td style="width:155px;">TM-321</td>
			<td style="width:297px;">Make sterilized distilled deionized water (ddH2O)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">P20 pipetman</td>
			<td style="width:243px;">Gilson</td>
			<td style="width:155px;">F123600</td>
			<td style="width:297px;">Add PPN into bee larval food pool</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:193px;">Incubator&#160;</td>
			<td style="width:243px;">Yihder Co., Ltd.</td>
			<td style="width:155px;">LE-550RD</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:193px;">Kimwipes</td>
			<td style="width:243px;">COW LUNG INSTRUMENT CO., LTD</td>
			<td style="width:155px;">KCS34155</td>
			<td style="width:297px;">Rearing pupae from extraction</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:193px;">Royal jelly</td>
			<td style="width:243px;">National Ilan University (NIU)</td>
			<td style="width:155px;">NIU</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">D-(+)-Glucose</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:155px;">G8270</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">D-(-)-Fructose</td>
			<td style="width:243px;">Sigma</td>
			<td style="width:155px;">F0127</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Yeast extract</td>
			<td style="width:243px;">CONDA, pronadisa</td>
			<td style="width:155px;">1702</td>
			<td style="width:297px;">Make basic larval diet (BLD)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:193px;">Sucrose</td>
			<td style="width:243px;">Taiwan sugar coporation</td>
			<td style="width:155px;">E01071010</td>
			<td style="width:297px;">Make sugar syrup for bee food</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:193px;">Pyriproxyfen (11%)</td>
			<td style="width:243px;">LIH-NUNG CHEMICAL CO.. LTD.</td>
			<td style="width:155px;">Registration No. 1937</td>
			<td style="width:297px;">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.

For the honey bee field test, a queen was limited to the 4-frame section for laying eggs. This step could increase the brood density in one frame and facilitate subsequent observations. Each treatment was labelled, and the honey bees&#39; development was clearly observed through a transparent slide. In vivo feeding of PPN-BLD to honey bee larvae in the beehive was performed to precisely evaluate the influence of PPN on the development of honey bees in the colony. Using the in...

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

Herein we present a method to feed pesticide contaminated food to both an individual honey bee and a beehive colony. The procedure evaluates the pesticide effect on individual honey bees by in vivo feeding of basic larval diet and also on the natural condition of beehive colony.

Het evalueren van het effect van Environmental Chemicals over Honey Bee ontwikkeling van het individu naar 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 ...

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

Geautomatiseerde Gel Size Selectie om de kwaliteit van next-generation sequencing bibliotheken Bereid uit Environmental watermonsters Verbeteren

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

Exploring the Effects of Atmospheric Forcings on Evaporation: Experimental Integration of the Atmospheric Boundary Layer and Shallow Subsurface

Het verkennen van de gevolgen van atmosferische forcering bij verdamping: Experimentele Integratie van de atmosferische grenslaag en ondiepe ondergrond

Evaporation is directly influenced by the interactions between the atmosphere, land surface and soil subsurface. This work aims to experimentally study ...

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Verwijdering van Trace Elements van Cupric Oxide Nanodeeltjes van Uranium<em&gt; In Situ</em&gt; Herstel Bleed Water en het effect op levensvatbaarheid van de cellen

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

Een flow-through Exposure System voor het evalueren van sedimenten in suspensie Effecten op Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Extraction of Organochlorine Pesticides from Plastic Pellets and Plastic Type Analysis

Extractie van organische chloride pesticiden uit plastic pellets en plastic type analyse

Plastic resin pellets, categorized as microplastics (&#8804;5 mm in diameter), are small granules that can be unintentionally released to the environment ...

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Preparation of DMMTAV and DMDTAV Using DMAV for Environmental Applications: Synthesis, Purification, and Confirmation

Voorbereiding van DMMTAV DMDTAV met DMAV voor milieutoepassingen: synthese, zuivering en bevestiging

Dimethylated thioarsenicals such as dimethylmonothioarsinic acid (DMMTAV) and dimethyldithioarsinic acid (DMDTAV), which are produced by the metabolic ...

The Effect of the Application of Thyme Essential Oil on Microbial Load During Meat Drying

Het Effect van de toepassing van essentiële tijmolie op microbiële laden tijdens het drogen van vlees

Meat is a high protein meal that is used in the preparation of jerky, a popular food snack, where preservation and safety are important. To assure food ...

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

Protocol voor Acute en chronische ecotoxiciteit testen van de Turquoise Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity ...

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee (Apis mellifera) Workers

Measuring Hypopharyngeal Gland Acinus Size in Honey Bee Apis mellifera Workers

Meten van de Hypopharyngeal de grootte van de Acinus van de klier in de honingbij (Apis mellifera) werknemers

The nurse hypopharyngeal glands produce the protein fraction of the worker and royal jelly that is fed to developing larvae and queens. These paired ...

Composition and Distribution Analysis of Bioaerosols Under Different Environmental Conditions

Samenstelling en analyse van de verdeling van Bioaerosols onder verschillende omgevingsomstandigheden

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