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Quantitative real‐time polymerase chain reaction (qPCR) is a method widely used to determine changes and differences in gene expression. As target gene expression is most often quantified relative to the expression of reference genes, the validation of suitable reference genes is of critical importance. In practice, however, such validation might not be thoroughly conducted if the same species and the same tissue or body parts are used for qPCR experiments. Here we show, that qPCR reference genes published for workers of European honey bee (Apis mellifera) subspecies fail to be stably expressed in workers of the African subspecies Apis mellifera scutellata. This is the case even when the sampled workers are in the same life stage, the same organ was dissected and the same reagents were used. Thus, reference genes need to be thoroughly re‐tested before they can be used as suitable references even when the only thing that changes is the subspecies used. [ABSTRACT FROM AUTHOR]

Background: Tsetse-transmitted African trypanosomiasis is a debilitating and fatal disease of humans and livestock if left untreated. While knowledge of the spatial distribution patterns of tsetse is essential for the development of risk-based vector control strategies, existing distribution maps in Zambia are more than 40 years old and were based on coarse spatial resolution data. The recently developed vehicle-mounted sticky trap (VST) provides an alternative sampling device to aid in updating existing distribution maps but has not been applied outside an experimental setting and is limited to motorable tracks. Therefore, the objective of the present study was to demonstrate the effectiveness of utilizing the VST for area-wide surveys of Glossina morsitans and to use the occurrence records to predict its spatial distribution in Zambia under current environmental conditions using Maxent. Methodology/Principal findings: Two-sided all-blue VST baited with butanone and 1-octen-3-ol was used to survey 692 and 1020 km of transect routes in G. m. centralis Machado and G. m. morsitans Westwood previously published distribution in Zambia. Maxent species distribution technique was used to predict the potential distribution of the two subspecies using current climatic and environmental data which was then compared to the historical distribution. A total of 15,602 tsetse were captured with G. m. morsitans (58%) being the most abundant. G. m. centralis and G. pallidipes Austin represented 39 and 2% of the catch respectively, and G. brevipalpis Newstead was also detected. The predicted potential distribution for G. m. centralis was 80,863 km2 while that of G. m. morsitans was 70,490 km2 representing a 47 and 29% reduction compared to their historical distributions, respectively. Conclusion/Significance: The VST is effective for sampling G. morsitans outside experimental settings and is recommended for use as an additional tsetse survey tool. The spatial distribution of G. morsitans in Zambia has reduced by 101,051 km2 due to temperature and land cover changes. Author summary: Tsetse habitat is increasingly fragmented owing to anthropogenic pressures and climate shifts which leads to spatial variability of African trypanosomiasis burden. Mapping tsetse occurrence is therefore a useful predictor of disease risk and provides the basis of evidence-based vector control strategies. In this study, we demonstrate the suitability of using the recently developed vehicle-mounted sticky trap for the rapid generation of occurrence records that can be used in ecological niche models to predict the distribution of G. morsitans. Since resources for vector surveillance and control are generally scarce in endemic settings, this alternative survey approach may be more efficient in the identification of priority areas for vector intervention. [ABSTRACT FROM AUTHOR]

Bee lice (Braulidae) are small parasitic flies, which are adapted to live on their bee host. As such, the wingless Braula coeca is a parasite of the common honey bee Apis mellifera and it is well adapted to attach to its hairy surface. The attachment system of B. coeca provides a secure grip on the fine setae of the bee. This is crucial for the parasite survival, as detachment from the host is fatal for the bee louse. The feet morphology of B. coeca is well adapted to the challenging bee surface, notably by strongly broadened claws, which are split into a high number of comb‐like teeth, perfectly matching the diameter of the bee hairs. Based on microscopy observations, both the morphology and material composition of the tarsi of B. coeca are characterized in detail. Using high‐speed video analysis, we combine the morphology data on the attachment system with a behavioural context. Furthermore, we directly measured the attachment forces generated by the bee lice in contact with the host. In particular, the claws are involved in attachment to the host, as the interstices between the teeth‐like spines allow for the collection of several hairs and generate strong friction, when the hairs slip to the narrow gap between the spines. The overall morphology of the tarsus produces strong attachment, with average safety factors (force per body weight) around 1130, and stabilizes the tarsal chain with lateral stoppers against overflexion, but also allows for the fast detachment by the tarsal chain torsion. [ABSTRACT FROM AUTHOR]

The African citrus triozid (ACT), Trioza erytreae, is an important pest of citrus. Both nymphs and adults damage the plant by feeding on the sap causing young shoots to die. Trioza erytreae also vectors Candidatus Liberibacter africanus, the bacteria that cause citrus greening disease. Since certain non‐host plants are known to repel insect pests, it is important to investigate how such plants can be exploited to manage T. erytreae. Here, we screened effects of odours of three non‐host plants namely guava (Psidium guajava), garlic (Allium sativum) and lemongrass (Cymbopogon citratus) against T. erytreae's location of a common host plant, rough lemon (Citrus jambhiri) and showed that repellence varied interspecifically with the plants. Using cage assays, we found that guava and garlic decreased the attraction of females but not males of T. erytreae to rough lemon volatiles. Chemical analysis by coupled gas chromatography/mass spectrometry (GC/MS) showed that volatiles of three of the plants were dominated by terpenoids; guava (69% comprised of limonene, 34%, (E)‐β‐ocimene, 29% and (Z)‐β‐Ocimene, 6%), lemongrass (56% comprised of geranial, 26%, neral, 19% and myrcene, 11%) and rough lemon (74% comprised of limonene, 53%, sabinene, 11% and (E)‐β‐ocimene, 10%). On the other hand, the volatile profile of garlic was dominated by benzenoids and saturated compounds (85% comprised of benzaldehyde, 12%, benzyl alcohol, 17%, nonanal, 31%, decanal, 13% and hexadecane, 12%). Our results suggest that non‐host plant volatile composition and richness in specific compounds may contribute to influencing T. erytreae response to its host, with garlic and guava as potential non‐host plants that can be exploited in the management of the pest. [ABSTRACT FROM AUTHOR]