Survival of honey bee (Apis mellifera) pupae after trypan blue staining of wounds caused by Varroa destructor mites or artificial perforation

Abstract

-The recently described technique of vital staining with trypan blue to visualise pupal wounds of honey bees, originating from punctures made by Varroa destructor mites, was applied to artificial perforations performed with a fine needle. The stained pupae were subsequently reared in vitro until eclosion of the adult bees. Their mortality was recorded daily. The survival of the treated pupae was only moderately affected by the staining procedure. No obvious toxic effects caused by the compound trypan blue were observed. The process of wound healing was normal. Pupae with previously trypan blue stained integumental wounds can therefore be used for long lasting experiments. This is of particular interest for future studies on the assumed vector function of Varroa mites in the transfer of bacterial and viral pathogens in honey bee diseases.Apis mellifera / Varroa destructor / pupal wounds / vital staining / trypan blue / survival

Immunogene and viral transcript dynamics during parasitic Varroa destructor mite infection of developing honey bee (Apis mellifera) pupae

“…Although puncturing the developing honey bee with a small glass capillary has been reported to inflict wounds similar to mite feeding sites (Herrmann et al, 2005), our data show profound, long-lasting perturbations of the expression of the investigated genes. The patterns of ∆C t values of immune genes relative to each other (supplementary material Fig.…”

“…Negative control larvae were left undisturbed. Larvae in a second control group received a small physical wound between the second thoracic segment and the first abdominal segment with a single-use capillary needle pulled to 50 μm diameter (Herrmann et al, 2005). Individuals in the three mite treatment groups received one, two, or three to five (3+) phoretic V. destructor mites.…”

Brood Affects Hygienic Behavior in the Honey Bee (Hymenoptera: Apidae)

“…Mites that could not clutch the paintbrush bristles were considered to be unhealthy, and were not used. Wounds that mimic Varroa mite feeding were inflicted with a 50-μm-diameter capillary needles on the dorsal side of the brood between the first abdominal segment and the second thoracic segment according to existing protocols (Herrmann et al 2005, Dade 2009. This wound was nonlethal, distinct from the pin-killed brood bioassay sometimes used for selection of hygienic behavior (Newton and Ostasiewski 1986).…”

Using an in vitro system for maintaining Varroa destructor mites on Apis mellifera pupae as hosts: studies of mite longevity and feeding behavior

“…Varroa females have serrated mouthparts that may be adapted for cutting through the cuticle of the honeybee host (Griffiths 1987). Using their mouthparts, mites create relatively large feeding holes in a host's integument, and these wounds have delayed healing (Donze and Guerin 1994;Herrmann et al 2005;Engels 2003, 2004). Varroa mites maintained on a honey bee pupa may feed from a single hole or multiple feeding holes Engels 2004, 2005).…”