Optimisation of an in vitro larvae test to test the effects of plant protection products on honeybee brood (Apis mellifera L.)

Lüken, Dorothee; Janke, Martina; Lienau, Friedrich-Wilhelm; Gerdel, Katharina; Ohe, Werner von der; Forster, Rolf

doi:10.1007/s00003-014-0865-0

Cited by 7 publications

(5 citation statements)

References 6 publications

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“…Otherwise, the data generated from those studies would not be considered valid . Previously published studies have noted higher control mortality rates than found in this study, thus demonstrating the reliability that the Schmehl et al in vitro rearing method offers …”

Section: Discussionsupporting

confidence: 79%

Acute toxicity of five pesticides to Apis mellifera larvae reared in vitro

Dai

Jack

Mortensen

et al. 2017

Pest Management Science

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Section: Discussionsupporting

confidence: 79%

Acute toxicity of five pesticides to Apis mellifera larvae reared in vitro

Dai

Jack

Mortensen

et al. 2017

Pest Management Science

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“…In our experience, this would translate to a poor rate of adult emergence given the already high rate of larval death which usually correlates with an equally high or higher rate of death in developing pupae. Other investigators experienced up to 60% mortality in controls prior to reaching adult emergence (Lüken et al, 2014). The reasons for low control survival and inconsistent results across different laboratories are unclear.…”

Section: Introductionmentioning

confidence: 99%

Protocol for the in vitro rearing of honey bee (Apis mellifera L.) workers

Schmehl

Tomé

Mortensen

et al. 2016

Journal of Apicultural Research

107

145

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The in vitro rearing of worker honey bees (Apis mellifera L.) has become an increasingly important method in honey bee research in general, and in pesticide risk assessment specifically. Authorities from the European Organization for Economic Co-operation and Development and the United States Environmental Protection Agency are requesting data on pesticide impacts on immature bee survivorship prior to registering new crop protection products. Those using the current in vitro rearing protocols have had variable success with immature bee survival and protocol repeatability. Here, we present an improved method for the in vitro rearing of worker honey bees from larvae to adult emergence. We have achieved consistently high survival (>95%) in our control and solvent-control rearing trials. Changes in the proportion of diet components, royal jelly source, maintenance of the developing bee, and rearing environment are the main contributors for our high rearing success and are discussed herein. Our in vitro rearing protocol can be implemented as the standard protocol to determine the impact of pesticides on immature bees because of the protocol's high control survivability, ease in end point determination, and high overall repeatability. Protocolo para la cría in vitro de obreras de Apis melliferaLa cría in vitro de las abejas obreras (Apis mellifera L.) se ha convertido en un método cada vez más importante en la investigació n de la abeja de la miel en general, y específicamente, en la evaluació n del riesgo de los plaguicidas. Las autoridades de la Organizació n Europea para la Cooperació n y el Desarrollo, y la Agencia de Protecció n Ambiental de los Estados Unidos están solicitando datos sobre los efectos de los plaguicidas en la supervivencia de abejas inmaduras antes de registrar nuevos productos para la protecció n de los cultivos. Aquellos que utilizan actualmente los protocolos de la cría in vitro han tenido un éxito variable en la supervivencia de abejas inmaduras y la repetitividad del protocolo. Aquí presentamos un método mejorado para la cría in vitro de las abejas obreras a partir de larvas para la eclosió n de los adultos. Hemos logrado consistentemente una alta supervivencia (>95%) en nuestro control y en los ensayos de cría con el disolvente del control. Los cambios en la proporció n de los componentes de la dieta, la fuente de jalea real, el mantenimiento de las abejas en desarrollo, y el ambiente de cría son los principales constituyentes para nuestro alto éxito de cría y son discutidos en este trabajo. Nuestro protocolo de cría in vitro puede ser implementado como el protocolo estándar para determinar el impacto de los plaguicidas en las abejas inmaduras debido a la alta capacidad de supervivencia del control, la facilidad en la determinació n del punto final, y la alta capacidad de repetitividad.

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“…In recent years, most labs use the method described by Aupinel et al (2005) and Aupinel et al (2007) . In a ring test that was carried out at seven laboratories, a high level of standardization of this method was attained ( Aupinel et al, 2009 ) and since then different groups have sought to further improve the protocol ( Crailsheim et al, 2013 ; Hendriksma, Härtel & Steffan-Dewenter, 2011a ; Lüken et al, 2014 ). However, despite the effort to standardize rearing protocols, surprisingly few studies exist that evaluate the ‘quality’ of bees obtained via this artificial rearing method.…”

Section: Introductionmentioning

confidence: 99%

Learning performance and brain structure of artificially-reared honey bees fed with different quantities of food

Steijven

Spaethe

Steffan‐Dewenter

et al. 2017

PeerJ

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BackgroundArtificial rearing of honey bee larvae is an established method which enables to fully standardize the rearing environment and to manipulate the supplied diet to the brood. However, there are no studies which compare learning performance or neuroanatomic differences of artificially-reared (in-lab) bees in comparison with their in-hive reared counterparts.MethodsHere we tested how different quantities of food during larval development affect body size, brain morphology and learning ability of adult honey bees. We used in-lab rearing to be able to manipulate the total quantity of food consumed during larval development. After hatching, a subset of the bees was taken for which we made 3D reconstructions of the brains using confocal laser-scanning microscopy. Learning ability and memory formation of the remaining bees was tested in a differential olfactory conditioning experiment. Finally, we evaluated how bees reared with different quantities of artificial diet compared to in-hive reared bees.ResultsThorax and head size of in-lab reared honey bees, when fed the standard diet of 160 µl or less, were slightly smaller than hive bees. The brain structure analyses showed that artificially reared bees had smaller mushroom body (MB) lateral calyces than their in-hive counterparts, independently of the quantity of food they received. However, they showed the same total brain size and the same associative learning ability as in-hive reared bees. In terms of mid-term memory, but not early long-term memory, they performed even better than the in-hive control.DiscussionWe have demonstrated that bees that are reared artificially (according to the Aupinel protocol) and kept in lab-conditions perform the same or even better than their in-hive sisters in an olfactory conditioning experiment even though their lateral calyces were consistently smaller at emergence. The applied combination of experimental manipulation during the larval phase plus subsequent behavioral and neuro-anatomic analyses is a powerful tool for basic and applied honey bee research.

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Optimisation of an in vitro larvae test to test the effects of plant protection products on honeybee brood (Apis mellifera L.)

Cited by 7 publications

References 6 publications

Acute toxicity of five pesticides to Apis mellifera larvae reared in vitro

Acute toxicity of five pesticides to Apis mellifera larvae reared in vitro

Protocol for the in vitro rearing of honey bee (Apis mellifera L.) workers

Learning performance and brain structure of artificially-reared honey bees fed with different quantities of food

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