The use of honey bees (Apis mellifera L.) to monitor airborne particulate matter and assess health effects on pollinators.

The honey bee Apis mellifera has long been recognized as an ideal bioindicator for environmental pollution. These insects are exposed to pollutants during their foraging activities, making them effective samplers of environmental contaminants, including heavy metals, pesticides, radionuclides, and volatile organic compounds. Recently, it has been demonstrated that honey bees can be a valuable tool for monitoring and studying airborne PM pollution, a complex mixture of particles suspended in the air, known to have detrimental effects on human health. Airborne particles attached to the bees can be characterised for their morphology, size, and chemical composition using a scanning electron microscopy coupled with X-ray spectroscopy, thus providing key information on the emission sources of the particles, their environmental fate, and the potential to elicit inflammatory injury, oxidative damage, and other health effects in living organisms. Here, we present a comprehensive summary of the studies involving the use of honey bees to monitor airborne PM, including the limits of this approach and possible perspectives. The use of honey bees as a model organism for ecotoxicological studies involving pollutant PM is also presented and discussed, further highlighting the role of the bees as a cornerstone of human, animal, and environmental health, according to the principles of the "One Health" approach.

Origin of non-exhaust PM in cities by individual analysis of particles collected by honey bees (Apis mellifera).

Urban areas present multiple challenges to scientists interested in unraveling the source, transport, and fate of airborne particulate matter (PM). Airborne PM consists of a heterogeneous mixture of particles with different sizes, morphologies, and chemical compositions. However, standard air quality stations only detect the mass concentration of PM mixtures with aerodynamic diameters ≤10 µm (PM10) and/or ≤ 2.5 µm (PM2.5). During honey bee foraging flights, airborne PM up to 10 µm in size attaches to their bodies, making them suitable for collecting spatiotemporal data on airborne PM. The individual particulate chemistry of this PM can be assessed using scanning electron microscopy coupled with energy-dispersive X-ray spectroscopy on a sub-micrometer scale, allowing accurate identification and classification of the particles. Herein, we analyzed the PM fractions of 10-2.5 µm, 2.5-1 µm, and below 1 µm in average geometric diameter collected by bees from hives located in the city of Milan, Italy. Bees showed contamination by natural dust, originating from soil erosion and rock outcropping in the foraging area, and particles with recurrent heavy metal content, most likely attributed to vehicular braking systems and possibly tires (non-exhaust PM). Notably, approximately 80% of non-exhaust PM was ≤1 µm in size. This study provides a possible alternative strategy to apportion the finer fraction of PM in urban areas and determine citizens' exposure. Our findings may also prompt decision-makers to issue policy addressal for non-exhaust pollution, especially for the ongoing restructuring of European regulations on mobility and the shift toward electric vehicles whose contribution to PM pollution is debated.

The Honey Bee Apis mellifera: An Insect at the Interface between Human and Ecosystem Health.

The concept of ecosystem services is widely understood as the services and benefits thatecosystems provide to humans, and they have been categorised into provisioning, regulating, supporting, and cultural services. This article aims to provide an updated overview of the benefits that the honey bee Apis mellifera provides to humans as well as ecosystems. We revised the role of honey bees as pollinators in natural ecosystems to preserve and restore the local biodiversity of wild plants; in agro-ecosystems, this species is widely used to enhance crop yield and quality, meeting the increasing food demand. Beekeeping activity provides humans not only with high-quality food but also with substances used as raw materials and in pharmaceuticals, and in polluted areas, bees convey valuable information on the environmental presence of pollutants and their impact on human and ecosystem health. Finally, the role of the honey bee in symbolic tradition, mysticism, and the cultural values of the bee habitats are also presented. Overall, we suggest that the symbolic value of the honey bee is the most important role played by this insect species, as it may help revitalise and strengthen the intimate and reciprocal relationship between humans and the natural world, avoiding the inaccuracy of considering the ecosystems as mere providers of services to humans.

Disentangling multiple PM emission sources in the Po Valley (Italy) using honey bees.

Particulate matter (PM) is a complex mixture of airborne chemical compounds commonly classified by their aerodynamic diameter. Although PM toxicity strongly depends on the morphology, chemical composition, and dimensions of particles, exposure limits set by environmental organisations only refer to the mean mass concentration of PM sampled daily or annually by monitoring stations. In this study, we used honey bees as sensors of airborne PM10 and PM2.5 in a highly polluted area of the Po Valley, northern Italy. Honey bees are an efficient sampler of airborne PM because, during flight and foraging activities, their pubescence promotes the accumulation of electrical charge on the body surface owing to air resistance, thus enhancing airborne PM attraction. Particles attached to the body of bees are readily accessible for physico-chemical characterisation using a scanning electron microscope coupled with X-ray spectroscopy (SEM/EDX). Our results demonstrate that residents in the study area are intermittently but chronically exposed to a well-defined spectrum of metal-bearing particles and mineral phases known to induce specific health outcomes. The morphology, size, and chemical composition of PM10 and PM2.5 detected on bees in the monitoring area were indicative of traffic, agricultural operations, and high-temperature combustion processes. The contribution of the A1 Milano-Bologna highway, local wheat and alfalfa cultivation, and the Parma incineration plant were clearly distinguishable. Our data also demonstrated that PM exposure levels may vary sharply throughout the year based on recurrent local activities.

Vehicle-derived ultrafine particulate contaminating bees and bee products.

Despite adverse health effects, ultrafine particulate matter (UFP), i.e., PM less than 0.1 µm in diameter, is an emerging pollutant not subject to regulation. UFP may cause both lung inflammation and cardiopulmonary disease and may enter the brain directly via the olfactory bulb, affecting the nervous system. In highly urbanized environments, diesel and gasoline vehicles are among the major sources of UFP including combustion-generated solid particle pollutant and metal-based particles. Metal-based UFP are of much concern, as they may promote inflammation and DNA damage via oxidative stress with generation of free radicals and reactive oxygen species (ROS). We used the honeybee as an alternative sampling system of UFP in an area of the Po Valley (Northern Italy), which is subject to intense traffic. Worker bees are widely recognised as efficient samplers of air pollutants, including airborne PM. During flight and foraging activity, pubescence of the bees promotes the accumulation of electrical charge on the body's surface, enhancing attraction to air pollutants. Bees living near the main Italian highway, the Autostrada A1, displayed a contamination of nanosized Fe-oxides/hydroxides and baryte. Sources of Fe-bearing and baryte ultrafine particles are primarily the vehicles speeding on the motorway. Pollen collected by forager bees and honey produced by the bee colony displayed contamination by nanosized Fe-oxides/hydroxides and baryte. Such a contamination exposes pollinators and humans to UFP ingestion, endangering the safety of food produced at traffic-influenced sites. Given the global spread of traffic, our findings suggest that exposure and environmental impact of ultrafine Fe-oxides/hydroxides and baryte are potentially ubiquitous, although usually overlooked in environmental policy discussions.

Particulate matter collection by honey bees (Apis mellifera, L.) near to a cement factory in Italy.

Industrial activities play a key role in the economic well-being of a country but they usually involve processes with a more or less profound environmental impact, including emission of pollutants. Among them, much attention has been given to airborne particulate matter (PM) whose exposure is ubiquitous and linked with several adverse health effects mainly due to its size and chemical composition. Therefore, there is a strong need to exploit monitoring systems for airborne PM able to provide accurate information on the potential health hazards and the specific emission sources for the implementation of adequate control strategies. The honey bee (Apis mellifera, L.) is widely used as an indicator of environmental pollution: this social hymenopteran strongly interacts with vegetables, air, soil, and water surrounding the hive and, as a consequence, pollutants from these sources are translated to the insect and to the hive products. During the wide-ranging foraging activity, the forager bee is known to collect samples of the main airborne PM pollutants emitted from different sources and therefore it can be used as an efficient PM sampler. In the present research, PM contaminating forager bees living nearby a cement factory and several kilometers away from it has been analysed and characterised morphologically, dimensionally and chemically through SEM/EDX. This provided detailed information on the role of both the cement manufacturing activities and the vehicular traffic as sources of airborne PM. This may indeed help the implementation of appropriate preventive and corrective actions that would effectively minimize the environmental spread of pollutant PM not only in areas close to the plant, but also in more distant areas.

Whole mitochondrial genomes unveil the impact of domestication on goat matrilineal variability.

BACKGROUND: The current extensive use of the domestic goat (Capra hircus) is the result of its medium size and high adaptability as multiple breeds. The extent to which its genetic variability was influenced by early domestication practices is largely unknown. A common standard by which to analyze maternally-inherited variability of livestock species is through complete sequencing of the entire mitogenome (mitochondrial DNA, mtDNA). RESULTS: We present the first extensive survey of goat mitogenomic variability based on 84 complete sequences selected from an initial collection of 758 samples that represent 60 different breeds of C. hircus, as well as its wild sister species, bezoar (Capra aegagrus) from Iran. Our phylogenetic analyses dated the most recent common ancestor of C. hircus to ~460,000 years (ka) ago and identified five distinctive domestic haplogroups (A, B1, C1a, D1 and G). More than 90 % of goats examined were in haplogroup A. These domestic lineages are predominantly nested within C. aegagrus branches, diverged concomitantly at the interface between the Epipaleolithic and early Neolithic periods, and underwent a dramatic expansion starting from ~12-10 ka ago. CONCLUSIONS: Domestic goat mitogenomes descended from a small number of founding haplotypes that underwent domestication after surviving the last glacial maximum in the Near Eastern refuges. All modern haplotypes A probably descended from a single (or at most a few closely related) female C. aegagrus. Zooarchaelogical data indicate that domestication first occurred in Southeastern Anatolia. Goats accompanying the first Neolithic migration waves into the Mediterranean were already characterized by two ancestral A and C variants. The ancient separation of the C branch (~130 ka ago) suggests a genetically distinct population that could have been involved in a second event of domestication. The novel diagnostic mutational motifs defined here, which distinguish wild and domestic haplogroups, could be used to understand phylogenetic relationships among modern breeds and ancient remains and to evaluate whether selection differentially affected mitochondrial genome variants during the development of economically important breeds.

Honey Bees (Apis mellifera, L.) as Active Samplers of Airborne Particulate Matter.

Honey bees (Apis mellifera L.) are bioindicators of environmental pollution levels. During their wide-ranging foraging activity, these hymenopterans are exposed to pollutants, thus becoming a useful tool to trace the environmental contaminants as heavy metals, pesticides, radionuclides and volatile organic compounds. In the present work we demonstrate that bees can also be used as active samplers of airborne particulate matter. Worker bees were collected from hives located in a polluted postmining area in South West Sardinia (Italy) that is also exposed to dust emissions from industrial plants. The area is included in an official list of sites of national interest for environmental remediation, and has been characterized for the effects of pollutants on the health of the resident population. The head, wings, hind legs and alimentary canal of the bees were investigated with Scanning Electron Microscopy coupled with X-ray spectroscopy (SEM-EDX). The analyses pointed to specific morphological and chemical features of the particulate, and resulted into the identification of three categories of particles: industry-, postmining-, and soil-derived. With the exception of the gut, all the analyzed body districts displayed inorganic particles, mostly concentrated in specific areas of the body (i.e. along the costal margin of the fore wings, the medial plane of the head, and the inner surface of the hind legs). The role of both past mining activities and the industrial activity close to the study area as sources of the particulate matter is also discussed. We conclude that honey bees are able to collect samples of the main airborne particles emitted from different sources, therefore could be an ideal tool for monitoring such a kind of pollutants.

Genetic diversity in widespread species is not congruent with species richness in alpine plant communities.

The Convention on Biological Diversity (CBD) aims at the conservation of all three levels of biodiversity, that is, ecosystems, species and genes. Genetic diversity represents evolutionary potential and is important for ecosystem functioning. Unfortunately, genetic diversity in natural populations is hardly considered in conservation strategies because it is difficult to measure and has been hypothesised to co-vary with species richness. This means that species richness is taken as a surrogate of genetic diversity in conservation planning, though their relationship has not been properly evaluated. We tested whether the genetic and species levels of biodiversity co-vary, using a large-scale and multi-species approach. We chose the high-mountain flora of the Alps and the Carpathians as study systems and demonstrate that species richness and genetic diversity are not correlated. Species richness thus cannot act as a surrogate for genetic diversity. Our results have important consequences for implementing the CBD when designing conservation strategies.

Reticulate evolution in stick insects: the case of Clonopsis (Insecta Phasmida).

BACKGROUND: Phasmids show noteworthy abilities to overcome species-specific reproductive isolation mechanisms, including hybridization, polyploidy, parthenogenesis, hybridogenesis and androgenesis. From an evolutionary standpoint, such tangled reproductive interactions lead to the complex phyletic relationships known as "reticulate evolution". Moroccan stick insects of the genus Clonopsis include one bisexual (C. felicitatis) and two closely related parthenogenetic forms (C. gallica, C. soumiae), which represent a polyploid series in chromosome number, but with apparent diploid karyotypes. Moreover, two Clonopsis strains of ameiotic males have been described, C. androgenes-35 and C. androgenes-53. As a consequence, Clonopsis stick insects may have experienced complex micro-evolutionary events, which we try to disentangle in this study. RESULTS: Mitochondrial cox2 analysis supports a recent divergence of Clonopsis, while AFLPs evidence genetic differentiation not linked to karyotypes, so that parthenogenetic C. gallica and C. soumiae appear to be a mix of strains of polyphyletic origin rather than single parthenogenetic species. Moreover, an admixed hybrid origin seems to be confirmed for C. androgenes. CONCLUSION: On the whole, Clonopsis is an intriguing case of reticulate evolution. Actually, complex cladogenetic events should be taken into account to explain the observed genetic structure, including diploidization of polyploid karyotypes, possibly coupled with hybridization and androgenesis. We also proposed a "working hypothesis" to account for the observed data, which deserves further studies, but fits the observed data very well.

Sex and stripping: The key to the intimate relationship between Wolbachia and host?

Wolbachia pipientis is known to infect only arthropods and nematodes (mainly filarial worms). A unique feature shared by the two Phyla is the ability to replace the exoskeleton, a process known as ecdysis. This shared characteristic is thought to reflect a common ancestry. Arthropod moulting is induced by the steroid hormone 20-hydroxyecdysone (20E) and a role for ecdysteroids in nematode ecdysis has also been suggested. Removing Wolbachia from filarial worms impairs the host's development. From analyses of the genome of Wolbachia harbored by the filarial nematode Brugia malayi and that of its host, the bacterium may provide a source of heme, an essential component of cytochrome P450's that are necessary for steroid hormone biosynthetic pathways.In arthropods, Wolbachia is a reproductive manipulator, inducing various phenotypic effects that may be due to differences in host physiology, in particular, endocrine-related processes governing development and reproduction. Insect steroids have well-defined roles in the coordination of multiple developmental processes, and in adults they control important aspects of reproduction, including ovarian development, oogenesis, sexual behavior, and in some taxa vitellogenin biosynthesis.According to some authors ecdysteroids may also act as sex hormones. In insects sex differentiation is generally thought to be a strictly genetic process, in which each cell decides its own sexual fate based on its sex chromosome constitution, but, surprisingly, recent data demonstrate that in Drosophila sex determination is not cell-autonomous, as it happens in mammals. Thus the presence of signals coordinating the development of a gender-specific phenotype cannot be excluded.This could explain why Wolbachia interferes with insect reproduction; and also could explain why Wolbachia interferes with insect development.Thus, is "sex (=reproduction) and stripping (=ecdysis)" the key to the intimate relationship between Wolbachia and its host?

History or ecology? Substrate type as a major driver of spatial genetic structure in Alpine plants.

Climatic history and ecology are considered the most important factors moulding the spatial pattern of genetic diversity. With the advent of molecular markers, species' historical fates have been widely explored. However, it has remained speculative what role ecological factors have played in shaping spatial genetic structures within species. With an unprecedented, dense large-scale sampling and genome-screening, we tested how ecological factors have influenced the spatial genetic structures in Alpine plants. Here, we show that species growing on similar substrate types, largely determined by the nature of bedrock, displayed highly congruent spatial genetic structures. As the heterogeneous and disjunctive distribution of bedrock types in the Alps, decisive for refugial survival during the ice ages, is temporally stable, concerted post-glacial migration routes emerged. Our multispecies study demonstrates the relevance of particular ecological factors in shaping genetic patterns, which should be considered when modelling species projective distributions under climate change scenarios.

The mystery of Etruscan origins: novel clues from Bos taurus mitochondrial DNA.

The Etruscan culture developed in Central Italy (Etruria) in the first millennium BC and for centuries dominated part of the Italian Peninsula, including Rome. The history of the Etruscans is at the roots of Mediterranean culture and civilization, but their origin is still debated: local or Eastern provenance? To shed light on this mystery, bovine and human mitochondrial DNAs (mtDNAs) have been investigated, based on the well-recognized strict legacy which links human and livestock populations. In the region corresponding to ancient Etruria (Tuscany, Central Italy), several Bos taurus breeds have been reared since historical times. These breeds have a strikingly high level of mtDNA variation, which is found neither in the rest of Italy nor in Europe. The Tuscan bovines are genetically closer to Near Eastern than to European gene pools and this Eastern genetic signature is paralleled in modern human populations from Tuscany, which are genetically close to Anatolian and Middle Eastern ones. The evidence collected corroborates the hypothesis of a common past migration: both humans and cattle reached Etruria from the Eastern Mediterranean area by sea. Hence, the Eastern origin of Etruscans, first claimed by the classic historians Herodotus and Thucydides, receives strong independent support. As the Latin philosopher Seneca wrote: Asia Etruscos sibi vindicat (Asia claims the Etruscans back).

Ultrastructural and molecular identification of a new Rickettsia endosymbiont in the springtail Onychiurus sinensis (Hexapoda, Collembola).

In this paper we provide microscopic and molecular evidence for the presence of an endosymbiontic bacterium in male and female gonads of the soil arthropod Onychiurus sinensis. The sequence of the gene encoding for the 16S rRNA shows that the bacterium is a member of the genus Rickettsia, and some anomalies presumably associated with the presence of these microorganisms have been detected. Although the Rickettsia found in O. sinensis has the smallest genetic divergence with Rickettsia bellii, the phylogenetic analysis fails to find support for a sister-group relationship between these two species, rather suggesting that most Rickettsia species/strains isolated in various arthropods have rapidly evolved and diversified in what appears to be a sudden burst of evolution.