Het evalueren van het effect van Environmental Chemicals over Honey Bee ontwikkeling van het individu naar Colony Level
Summary
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.
Abstract
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.
Introduction
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.
Protocol
Representative Results
Discussion
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…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
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).
Materials
| Honey bee box | SAN-YI Honey Factory | W1266 | Honeybees rearing |
| Queen excluder (between frames) | SAN-YI Honey Factory | I1575 | Queen limitation |
| Queen excluder (on top ) | SAN-YI Honey Factory | I1566 | Queen limitation on top |
| Bee brush | SAN-YI Honey Factory, Taiwan | W1414 | clean the bees on frame gently |
| Bee feeder | SAN-YI Honey Factory, Taiwan | P0219 | feed sugar syrup to colony |
| Transparent slide | Wan-Shih-Chei, Taiwan (http://www.mbsc.com.tw/a01goods.asp?s_id=40) | 1139 | Mark the larval area on the frames (Material: Polyethylene Terephthalate, PET) (Size= Length*Width*thick= 29.7mm*21mm*0.1mm) |
| 24 well tissu culture plate | Guangzhou Jet Bio-Filtration Co., Ltd | TCP011024 | Rearing pupae from extraction |
| Autoclave | Tomin medical equipmenco., LTD. | TM-321 | Make sterilized distilled deionized water (ddH2O) |
| P20 pipetman | Gilson | F123600 | Add PPN into bee larval food pool |
| Incubator | Yihder Co., Ltd. | LE-550RD | Rearing pupae from extraction |
| Kimwipes | COW LUNG INSTRUMENT CO., LTD | KCS34155 | Rearing pupae from extraction |
| Royal jelly | National Ilan University (NIU) | NIU | Make basic larval diet (BLD) |
| D-(+)-Glucose | Sigma | G8270 | Make basic larval diet (BLD) |
| D-(-)-Fructose | Sigma | F0127 | Make basic larval diet (BLD) |
| Yeast extract | CONDA, pronadisa | 1702 | Make basic larval diet (BLD) |
| Sucrose | Taiwan sugar coporation | E01071010 | Make sugar syrup for bee food |
| Pyriproxyfen (11%) | LIH-NUNG CHEMICAL CO.. LTD. | Registration No. 1937 | Insect growth regulator (IGR) used in the experiment |
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