Abstract Rapid, accurate and cost-effective identification of Apis mellifera subspecies is needed for subspecies of regulatory concern. We designed and validated subspecies markers based on single nucleotide polymorphisms (SNP) on mitochondrial cytochrome b ( Cytb ) and NADH dehydrogenase 4 ( ND4 ) genes. We used a combination of established and novel real-time qPCRs in a stepwise series, with increasing discrimination power, to (1) differentiate honey bees of African (A-lineage) ancestry from those of other lineages ( Cytb SNP #1), (2) identify African-derived honey bees (AHBs) ( Cytb SNP #2), and (3) detect A. m. capensis exclusively in the bee’s indigenous region of South Africa ( ND4 SNP). We also developed a restriction fragment length polymorphism assay targeting a SNP on the NADH dehydrogenase 2 ( ND2 RFLP) gene to detect the specific mitochondrial A-lineage clade. These assays allow for reliable time- and cost-effective results that provide increased accuracy on subspecies assignation.

Pests and pathogens are a continuous threat to the health of Western honey bee Apis mellifera L. Monitoring honey bee colonies for arthropod pests, disease-causing bacteria and fungi, and early detection of new invasions is essential to maintain the pollination services provided by honey bees. We investigated the feasibility of using eDNA metabarcoding to detect pests and pathogens in bee hives and across their foraging environment. We sampled 13 sources for eDNA within and outside hives from our test apiary to determine where the most informative eDNA could be obtained, with most sources sampled thrice. This resulted in 61 samples, 20 negative controls, and three positive controls. Furthermore, we compared two eDNA collection techniques-wiping surfaces with moistened forensic swabs and using a spray/wash technique that aggregated surface eDNA into a container, before collecting the eDNA on a filter. We used DNA metabarcoding with universal primer sets to target arthropod, bacterial, and fungal communities. Our results showed that most sources yielded sufficient eDNA and that results of the swab and spray/wash methods were similar when they could be applied to the same surface. We detected DNA from honey bee bacterial symbionts, mycotoxin-producing fungi and Brachymyrmex sp. rover ants. Common pests and pathogens of concern to honey bees (i.e., [small hive beetle (Aethina tumida), Varroa destructor, and Melissococcus plutonius]) were detected. This matched our visual observations of clinical signs of these pests and pathogens in the hives we tested. DNA from some species was source specific, which has implications for using eDNA as a monitoring tool. Collectively, our data show that eDNA metabarcoding can accurately detect DNA from arthropods and microbes honey bees contact and can be used as a comprehensive molecular predictor tool for colony health surveys.

Low temperatures are rarely experienced in isolation. The impacts of low temperatures on insects can be exacerbated or alleviated by the addition of other environmental factors, including, for example, desiccation, hypoxia, or infection. One way in which environmental factors can interact is through cross-talk where different factors enact common signaling pathways. In this review, I highlight the breadth of abiotic and biotic factors that can interact with low temperature tolerance in both natural and artificial environments; and discuss some of the candidate pathways that are possibly responsible for cross-talk between several factors. Specifically, I discuss three interesting candidates: the neurohormone octopamine, circadian clock gene vrille, and microbes. Finally, I discuss applications of cross-talk studies, and provide recommendations for researchers.

BackgroundThe sterile insect technique (SIT) is emerging as a tool to supplement traditional pesticide-based control of Aedes aegypti, a prominent mosquito vector of microbes that has increased the global burden of human morbidity and mortality over the past 50 years. SIT relies on rearing, sterilizing and releasing large numbers of male mosquitoes that will mate with fertile wild females, thus reducing production of offspring from the target population. In this study, we investigated the effects of ionizing radiation (gamma) on male and female survival, longevity, mating behavior, and sterility of Ae. aegypti in a dose-response design. This work is a first step towards developing an operational SIT field suppression program against Ae. aegypti in St. Augustine, Florida, USA. ResultsExposing late-stage pupae to 50 Gy of radiation yielded 99% male sterility while maintaining similar survival of pupae to adult emergence, adult longevity and male mating competitiveness compared to unirradiated males. Females were completely sterilized at 30 Gy, and when females were dosed with 50 Gy, they had a lower incidence of blood-feeding than unirradiated females. ConclusionOur work suggests that an ionizing radiation dose of 50 Gy should be used for future development of operational SIT in our program area because at this dose males are 99% sterile while maintaining mating competitiveness against unirradiated males. Furthermore, females that might be accidentally released with sterile males as a result of errors in sex sorting also are sterile and less likely to blood-feed than unirradiated females at our 50 Gy dose. (c) 2022 Society of Chemical Industry.

Incorporating physiology into models of population dynamics will improve our understanding of how and why invasions succeed and cause ecological impacts, whereas others fail or remain innocuous. Targeting both organismal physiologists and invasion scientists, we detail how physiological processes affect every invasion stage, for both plants and animals, and how physiological data can be better used for studying the spatial dynamics and ecological effects of invasive species. We suggest six steps to quantify the physiological functions related to demography of nonnative species: justifying physiological traits of interest, determining ecologically appropriate time frames, identifying relevant abiotic variables, designing experimental treatments that capture covariation between abiotic variables, measuring physiological responses to these abiotic variables, and fitting statistical models to the data. We also provide brief guidance on approaches to modeling invasions. Finally, we emphasize the benefits of integrating research between communities of physiologists and invasion scientists.

A real-time qPCR assay was designed to detect African-derived subspecies of western honey bees (Apis mellifera L.). The probes targeted the same region of the mitochondrial cytochrome b gene used in previous restriction enzyme assays. Using samples from 16 A. mellifera subspecies representing four lineages, we evaluated the efficacy of this assay to identify African- (or A-lineage) and non-A-lineage honey bees. The qPCR assay successfully differentiated A-lineage honey bees, including A. m. scutellata and A. m. capensis, from M-, C-, and O-lineage A. mellifera subspecies. In conclusion, the assay we developed is useful for rapid detection of A-lineage honey bees in areas where they are introduced, although it cannot be used to identify the specific A. mellifera subspecies within the A-lineage.

The complete mitochondrial genome of Apis mellifera simensis was 16,523 bp long. The 13 protein-coding genes, two rRNAs, and 22 tRNAs resembled other Apis mitogenomes. The location of this Apis subspecies in our phylogenetic tree supported the hypothesis that this subspecies is distinct, and is most closely related to A. m. scutellata and A. m. monticola.

Aedes aegypti is a prominent disease vector that is difficult to control through traditional integrated vector management due to its cryptic peridomestic immature-stage habitat and adult resting behavior, increasing resistance to pesticide formulations approved by the US Environmental Protection Agency, escalating deregistration of approved pesticides, and slow development of new effective chemical control measures. One novel method to control Ae. aegypti is the sterile insect technique (SIT) that leverages the mass release of irradiated (sterilized) males to overwhelm mate choice of natural populations of females. However, one potential liability of SIT is sex sorting errors prior to irradiation, resulting in accidental release of females. Our goal in this study was to test the extent to which irradiation affects female life-history parameters to assess the potential impacts of releasing irradiated females accidentally sorted with males. In this study, we determined that a radiation dose ≥30 Gy-a dose sufficient to sterilize males while preserving their mating competitiveness-may substantially impact longevity, bloodfeeding, oviposition, and egg hatch rate of female Ae. aegypti after being irradiated as pupae. These findings could reduce public concern for accidental release of females alongside irradiated males in an operational Ae. aegypti SIT control program.

The mitochondrial genome of a worker Apis mellifera jemenitica was 16,623 bp. It consisted of 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs and a control region. Phylogenetic analyses suggest a close relationship between A. m. jemenitica, A. m. lamarckii and A. m. syriaca.

BACKGROUND Phytosanitary irradiation is a sustainable alternative to chemical fumigants for disinfesting fresh commodities from insect pests. However, irradiating insects in modified atmospheres with very low oxygen (<1 kPa O2 ) has repeatedly been shown to increase radioprotective response. Thus, there is a concern that modified atmosphere packaging could reduce the efficacy of phytosanitary irradiation. One hurdle slowing the widespread application of phytosanitary irradiation is a lack of knowledge about how moderate levels of hypoxia relevant to modified atmosphere packaging of most fresh commodities (3-10 kPa O2 ) may affect phytosanitary irradiation treatments. Therefore, we hypothesize that critical PO2 (Pcrit ), the level of oxygen at which an insect's metabolism becomes impaired, can be used as a diagnostic biomarker to predict the induction of a radioprotective response. RESULTS Using the cabbage looper Trichoplusia ni (Hübner), we show that there is a substantial increase in radiation resistance when larvae are irradiated in atmospheres more hypoxic than their Pcrit (3.3 kPa O2 ). These data are consistent with our hypothesis that Pcrit could be used as a diagnostic biomarker for what levels of hypoxia may induce radioprotective effects that could impact phytosanitary irradiation treatments. CONCLUSION We propose that the relationship between Pcrit and radioprotective effects could allow us to build a framework for predicting the effects of low-oxygen atmospheres on the efficacy of phytosanitary irradiation. However, more widespread studies across pest species are still needed to test the generality of this idea. This article is protected by copyright. All rights reserved.

The mitochondrial genome of Apis mellifera ruttneri consisted of 13 protein-coding genes, two rRNAs, 22 tRNAs, an AT-rich control region, and was 16,577 bp long. The phylogenetic analyses suggested that A. m. ruttneri was closely related to two North African subspecies: A. m. sahariensis and A. m. intermissa.

Apis mellifera anatoliaca had a mitochondrial genome that was 16,256 bp long, with 13 protein-coding genes, 22 tRNA genes, two rRNA genes, and an AT-rich control region. The phylogenetic tree showed that A. m. anatoliaca was closely related to other subspecies found in Turkey, A. m. caucasica and A. m. meda.

The complete mitochondrial genome of the West African honey bee Apis mellifera adansonii consisted of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region. It was 16,466 bp and consisted of 84.7% AT nucleotides. This subspecies had a similar mitogenome to those of other southern African honey bees, namely A. m. scutellata, A. m. capensis, and A. m. monticola.

Abstract The complete mitochondrial genome of the endemic Malagasy honey bee Apis mellifera unicolor is 16,373 bp and comprises 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a control region. The mitochondrial genome closely resembles mitogenomes of other published Apis mellifera subspecies, and the phylogenetic analysis suggests that A. m. unicolor is distinct from other African (A) lineage honey bees but is most closely related to the honey bees from southern African: A. m. scutellata and A. m. capensis.