SummaryOriginally, a method to rear worker honey bee larvae in vitro was introduced into the field of bee biology to analyse honey bee physiology and caste development. Recently, it has become an increasingly important method in bee pathology and toxicology. The in vitro method of rearing larvae is complex and can be developed as an art by itself, especially if the aim is to obtain queens or worker bees which, for example, can be re-introduced into the colony as able members. However, a more pragmatic approach to in vitro rearing of larvae is also possible and justified if the aim is to focus on certain pathogens or compounds to be tested. It is up to the researcher(s) to decide on the appropriate experimental establishment and design. This paper will help with this decision and provide guidelines on how to adjust the method of in vitro rearing according to the specific needs of the scientific project. Métodos estándar para la cría in vitro de larvas de Apis mellifera ResumenOriginalmente, el método para la cría in vitro de larvas de obreras de abejas melíferas se introdujo en el campo de la biología de las abejas para analizar la fisiología y el desarrollo de las castas. Recientemente, se ha convertido en un método cada vez más importante para la patología y la toxicología de la abeja. El método de cría in vitro de larvas es complejo y constituye un arte en sí mismo, especialmente si el objetivo es obtener reinas o abejas obreras que, por ejemplo, puedan ser re-introducidas en la colonia como miembros activos. Sin embargo, un enfoque más pragmático de la cría de larvas in vitro también es posible y justificado si el objetivo es centrarse en ensayos con ciertos patógenos o compuestos. Corresponde al investigador (es) decidir sobre el adecuado establecimiento experimental y el diseño. Este artículo ayudará con esta decisión y proporcionará directrices sobre cómo ajustar el método de cría in vitro en función de las necesidades específicas del proyecto científico.

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BackgroundDouble sensitization (DS) to bee and vespid venom is frequently observed in the diagnosis of hymenoptera venom allergy, but clinically relevant DS is rare. Therefore it is sophisticated to choose the relevant venom for specific immunotherapy and overtreatment with both venoms may occur. We aimed to compare currently available routine diagnostic tests as well as experimental tests to identify the most accurate diagnostic tool.Methods117 patients with a history of a bee or vespid allergy were included in the study. Initially, IgE determination by the ImmunoCAP, by the Immulite, and by the ADVIA Centaur, as well as the intradermal test (IDT) and the basophil activation test (BAT) were performed. In 72 CAP double positive patients, individual IgE patterns were determined by western blot inhibition and component resolved diagnosis (CRD) with rApi m 1, nVes v 1, and nVes v 5.ResultsAmong 117 patients, DS was observed in 63.7% by the Immulite, in 61.5% by the CAP, in 47.9% by the IDT, in 20.5% by the ADVIA, and in 17.1% by the BAT. In CAP double positive patients, western blot inhibition revealed CCD-based DS in 50.8%, and the CRD showed 41.7% of patients with true DS. Generally, agreement between the tests was only fair and inconsistent results were common.ConclusionBAT, CRD, and ADVIA showed a low rate of DS. However, the rate of DS is higher than expected by personal history, indicating that the matter of clinical relevance is still not solved even by novel tests. Furthermore, the lack of agreement between these tests makes it difficult to distinguish between bee and vespid venom allergy. At present, no routinely employed test can be regarded as gold standard to find the clinically relevant sensitization.

Immunological responses after stings varied in bee and vespid venom-allergic patients. In patients under VIT, sIgE and sIgG remained completely stable after sting challenges. Monitoring VIT efficacy was only possible in vespid venom allergy, and the sIgG threshold for rVes v 5 had the highest sensitivity to confirm tolerance. The BAT inhibition test was the most reliable tool to confirm tolerance on an individual basis.