Suitability of drone olfactory sensitivity as a selection trait for Varroa-resistance in honeybees

The most effective strategy against brood diseases, such as those stemming from infestation by the mite Varroa destructor, is the early detection and removal of sick brood. Recent findings suggest that genes associated with worker bee olfactory perception play a central role in Varroa-sensitive hygiene (VSH). In this study, the odour sensitivity of Apis mellifera drones was examined through proboscis extension response (PER) conditioning. Individuals sensitive/insensitive to the two Varroa-parasitised-brood odours (extract-low and extract-high) were used for breeding. Twenty-one queens from a VSH-selected line (SelQ) and nineteen queens from a nonselected line (ConQ) were single-drone-inseminated with sperm from drones that showed either sensitivity (SenD+) or insensitivity (SenD−) to the two extracts. Individual VSH behaviour in a total of 5072 offspring of these combinations (SelQ × SenD+, SelQ× SenD−, ConQ × SenD+, ConQ × SenD−) was subsequently observed in a specially designed observation unit with infrared light. The results from the video observation were also separately examined, considering the genetic origin (VSH-selected or nonselected line) of the participating queens and drones. While the drone PER conditioning results were not significantly reflected in the VSH results of the respective offspring, the genetic origin of the participating queens/drones was crucial for VSH manifestation.

qPCR assays with dual-labeled probes for genotyping honey bee variants associated with varroa resistance

Background The varroa mite is one of the main causes of honey bee mortality. An important mechanism by which honey bees increase their resistance against this mite is the expression of suppressed mite reproduction. This trait describes the physiological inability of mites to produce viable offspring and was found associated with eight genomic variants in previous research. Results This paper presents the development and validation of high-throughput qPCR assays with dual-labeled probes for discriminating these eight single-nucleotide variants. Amplicon sequences used for assay validation revealed additional variants in the primer/probe binding sites in four out of the eight assays. As for two of these the additional variants interfered with the genotyping outcome supplementary primers and/or probes were developed. Inclusion of these primers and probes in the assay mixes allowed for the correct genotyping of all eight variants of interest within our bee population. Conclusion These outcomes underline the importance of checking for interfering variants in designing qPCR assays. Ultimately, the availability of this assay allows genotyping for the suppressed mite reproduction trait and paves the way for marker assisted selection in breeding programs.

qPCR Assays with Dual-Labeled Probes for Genotyping Honey bee Variants Associated with Varroa Resistance

Background: The varroa mite is one of the main causes of honey bee mortality. An important mechanism by which honey bees increase their resistance against this mite is the expression of suppressed mite reproduction. This trait describes the physiological inability of mites to produce viable offspring and was found associated with eight genomic variants in previous research. Results: This paper presents the development and validation of high-throughput qPCR assays with dual-labeled probes for discriminating these eight single-nucleotide variants. Within the sequenced samples, additional variants were detected in the primer/probe binding sites in four out of the eight variants of interest. As for two of these the additional variants interfered with the genotyping outcome supplementary primers and/or probes were developed. Inclusion of these primers and probes in the assay mixes allowed for the correct genotyping of all eight variants of interest.Conclusion: These outcomes underline the importance of checking for interfering variants in designing qPCR assays. Ultimately, the availability of this assay allows genotyping for the suppressed mite reproduction trait and paves the way for marker assisted selection in breeding programs.

Evaluation of Traits for the Selection of Apis Mellifera for Resistance against Varroa Destructor

Infestation with Varroa destructor is a serious cause of bee colony (Apis mellifera) losses on a global level. However, the presence of untreated survivor populations in many different regions supports the idea that selection for resistance can be successful. As colony survival is difficult or impossible to measure, differences in mite infestation levels and tests for specific behavioral traits are used for selective breeding for Varroa resistance. In this paper we looked into different definitions of mite infestation and linked these with brood hygiene (pin test), brood recapping and suppressed mite reproduction. We based our analyses on datasets of Apis mellifera carnica from three countries: Austria (147 records), Croatia (135) and Germany (207). We concluded that bee infestation in summer, adjusted for the level of natural mite fall in spring, is a suitable trait in the breeding objective, and also suggested including brood infestation rate and the increase rate of bee infestation in summer. Repeatability for bee infestation rate was about 0.55, for cells opened in pin test about 0.33, for recapping 0.35 and for suppressed mite reproduction (SMR) virtually zero. Although in most cases we observed correlations with the expected sign between infestation parameters and behavioral traits, the values were generally low (<0.2) and often not significantly different from zero.

Descriptive Analysis of the Varroa Non-Reproduction Trait in Honey Bee Colonies and Association with Other Traits Related to Varroa Resistance

In the current context of worldwide honey bee colony losses, among which the varroa mite plays a major role, the hope to improve honey bee health lies in part in the breeding of varroa resistant colonies. To do so, methods used to evaluate varroa resistance need better understanding. Repeatability and correlations between traits such as mite non-reproduction (MNR), varroa sensitive hygiene (VSH), and hygienic behavior are poorly known, due to practical limitations and to their underlying complexity. We investigate (i) the variability, (ii) the repeatability of the MNR score, and (iii) its correlation with other resistance traits. To reduce the inherent variability of MNR scores, we propose to apply an empirical Bayes correction. In the short-term (ten days), MNR had a modest repeatability of 0.4, whereas in the long-term (a month), it had a low repeatability of 0.2, similar to other resistance traits. Within our dataset, there was no correlation between MNR and VSH. Although MNR is amongst the most popular varroa resistance estimates in field studies, its underlying complex mechanism is not fully understood. Its lack of correlation with better described resistance traits and low repeatability suggest that MNR needs to be interpreted cautiously, especially when used for selection.

Descriptive Analysis of the Varroa Non-reproduction Trait in Honey Bee Colonies and Association with Other Traits Related to Varroa Resistance

In the current context of worldwide honey bee colony losses, among which the varroa mite plays a major role, hope to improve honey bee health lies in part in the breeding of varroa resistant colonies. To do so, methods used to evaluate varroa resistance need better understanding. Repeatability and correlations between traits such as Mite Non-Reproduction (MNR), Varroa Sensitive Hygiene (VSH) and hygienic behaviour are poorly known, due to practical limitations and to their underlying complexity. We investigate (i) the variability, (ii) repeatability of the MNR score and (iii) its correlation with other resistance traits. To reduce the inherent variability of MNR scores, we propose to apply an Empirical Bayes correction. On the short-term (ten days) MNR had a modest repeatability of 0.4 whereas on the long- term (a month) it had a low repeatability of 0.2, similar to other resistance traits. Within our dataset there was no correlation between MNR and VSH. Although MNR is amongst the most popular varroa resistance estimates in field studies, its underlying complex mechanism is not fully understood. Its lack of correlation with better described resistance traits and low repeatability suggest that MNR need to be interpreted cautiously, especially when used for selection.