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Table 1.

Primer sequences used for real-time PCR of selected Varroa mite genes.

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Table 2.

Comparison of phoretic Varroa mite survival and feeding on media based on drone larva hemolymph or worker larva hemolymph with addition of ascorbic acid.

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Table 2 Expand

Table 3.

The comparison of various dyes included in the feeding medium to visually identify mites that had ingested media.

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Fig 1.

The visual detection of dyes in the Varroa mite after feeding.

Of the four dyes that were ingested, (A) the Royal blue was the easiest to visually detect in the mite alimentary tract. The (B) Indigo, (C) Neutral Red and (D) Red Bengal were more difficult to differentiate from normal colors of the mites. Arrows point to regions within the mites where ingested dyes could be observed.

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Fig 2.

The effect of frozen storage on the efficacy of drone larva hemolymph to support feeding and survival of phoretic Varroa mites.

Percentage of mites that fed and survived with hemolymph stored up to 6 weeks at -20°C was comparable to fresh hemolymph (F = 4.53, p = 0.0188, n = 4–6). Letter b designates statistical difference from a. Abbreviations: 3w = three weeks; 6w = six weeks; 9w = nine weeks.

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Fig 2 Expand

Fig 3.

Comparison of transcript levels for the constitutively expressed actin and GAPDH genes in phoretic mites and those surviving the feeding protocol.

Black bars are phoretic mites and gray bars represent mites completing feeding protocol. T bars represent standard error of the mean. Abbreviations: VdActin = actin and VdGAPDH = GAPDH.

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Fig 4.

Relative transcript levels of nine selected genes in phoretic mites and in mites that had completed the feeding protocol.

All transcript levels were normalized to the geometric mean of actin and GAPDH transcript levels and presented as fold differences as reported previously [59]. Black bars are phoretic mites and gray bars represent mites completing feeding protocol. T bars represent standard error of the mean. Abbreviations: LLTP = large lipid transport protein, Vg1 = vitellogenin 1, Vg2 = vitellogenin 2, For = foraging, Mvl = malvolio, Spo = spook, Dib = disembodied, Shd = shade and Sub = subolesin.

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Fig 5.

Elevation of selected transcripts by feeding on Tebufenozide.

Phoretic Varroa mites were fed on media containing varying concentrations of Tebufenozide in the feeding protocol. RT-qPCR was conducted on the total RNA extracts from the surviving mites. Transcript levels were determined for the (A) large lipid transport protein (LLTP), (B) vitellogenin 1 (Vg1), (C) vitellogenin 2 (Vg2), (D) disembodied (Dib), (E) shade (Shd) and (F) spook (Spo) genes and normalized with the geometric mean for actin and GAPDH as reported previously [59]. T bars represent standard error of the mean. Letter “b” in panel A and E designate significant statistical difference from treatments with “a”. No statistical designations were added to the other panels because there were no statistical differences between treatments.

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