Evaluering af effekten af Environmental Chemicals på Honey Bee Udvikling fra den enkelte til Colony Level
Summary
Heri præsenterer vi en fremgangsmåde til foder pesticid forurenede fødevarer til både en individuel honningbi og en bikube koloni. Proceduren evaluerer pesticidet virkning på individuelle honningbier ved in vivo tilførsel af basisk larvediæt og også på den naturlige tilstand bikube koloni.
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
Tilstedeværelsen af pesticider i miljøet er en af de mest alvorlige problemer, som påvirker livet for en honningbi 1, 2, 3. Flere undersøgelser har påvist den fælles tilstedeværelse af pesticidrester i honning bikolonier og biprodukter. I Taiwan, den gennemsnitlige anvendelse af pesticider var 11-12 kg / ha hvert år (fra 2005 til 2013). Mængden af pesticider, der anvendes i Taiwan er højere end i EU-landene, og de latinamerikanske lande 4, 5. Med andre ord, er fra stadet miljø lider alvorlig pesticid stress, især i Taiwan og muligvis i andre lande.
Honningbien Apis mellifera er en af de store bestøvere i landbrugssystemer 6 og det producerer også værdifulde produkter såsom honning. Men honningbier er udstillingered til forskellige pesticider og disse pesticider kan bringes tilbage til bistader efter fouragere på blomster, der er blevet sprøjtet med pesticider, når de indsamler nektar og pollen 7, 8. De kan også blive udsat for pesticider ved biavlerne selv har til formål at kontrollere problemer med skadedyr inde bistader 9, 10, 11. Fordi honningbi larver fodret med sygeplejerske bier for deres udvikling, larver, droner og endda dronningen kan udsættes for disse pesticider-kontamineret nektar og pollen 12. Toksiciteten af forskellige pesticider til honningbier skal løses 13.
Der er gjort mange forsøg på at vurdere spørgsmålene om miljømæssige pesticidrester. Yang et al. testet påvirkningen af det neurotoksiske insektmidlet imidacloprid på udviklingen af honningbi larver ibikube og rapporterede, at en sub-letal dosis af imidacloprid resulterede i olfaktoriske associative opførsel af de voksne bier 14. Også, Urlacher et al. undersøgte de subletale virkninger af et organophosphat pesticider, chlorpyrifos, på en honningbi arbejdstagers læring ydeevne under laboratorieforhold 15. I vores tidligere undersøgelse, vi evaluerede effekten af et insekt vækst regulator (IGR), pyriproxyfen (PPN), på larver honningbier 16.
I dette papir, vi præsenterer metoder til evaluering af de kemiske virkninger på udviklingen af honningbier. Honey bee fodringsmetoder blev beskrevet og anvendt til enten individuelle honningbier eller til en koloni. Først testede vi forskellige koncentrationer af pesticid-forurenet basisk larvediæt (BLD) på larver i kolonierne at vurdere virkningen af pesticidet på individuelle honningbier in vivo. Vi fortsatte derefter med at simulere den naturlige conditioner af pesticidet ved anvendelse pesticid-forurenet sirup i bistader. Ved denne fremgangsmåde vil PPN, som er meget udbredt mod skadedyr 17 og er skadeligt for udviklingen af honningbi-larver og pupper 16, 18, 19, være en indikator til at repræsentere den negative virkning af pesticidet i marken.
Protocol
Representative Results
Discussion
Dronningen-begrænset æglægning metode og dronning-udveksling metode er kritiske trin til opsætning af honningbi grupper til feltforsøg inden for denne protokol. Dronningen-begrænset æglægning metode tillader synkronisering af livscyklus honningbier. Derfor kan forskerne vælge 1 dag-gamle larver af samme alder for behandling med forskellige doser af pesticider. For queen-udveksling metode blev dronningen udveksles mellem del A (4 rammer) og B (5 billeder) for at opnå forskellige udviklingsstadier i honningbi fo…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
Denne forskning blev støttet af Grant 105AS-13.2.3-BQ-B1 fra Bureau of Animal og Plantesundhed Inspection and Quarantine, Rådet for landbrug, Executive Yuan og Grant 103-2313-B-197-002-MY3 fra ministeriet for Videnskab og Teknologi (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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