Rearing techniques for hornets with emphasis on Vespa velutina (Hymenoptera: Vespidae): A review

Of the 34 vespid species recorded worldwide as invasive aliens, particular attention is currently being given to the yellow-legged hornet Vespa velutina that has invaded Europe, Japan, and Korea. The hornet is a voracious predator of bees and a serious threat to bee colonies and bee pollination. Control measures are needed, but their development has been challenging as biological and ecological studies are limited by the short field season and the cryptic nesting behavior of these hornets. A Vespa rearing process can generate the large numbers of workers, gynes, and males that are essential for studying chemical ecology and life history and for experimental testing of various hypotheses. We present a synthesis of suitable methods and techniques for year-round Vespa hornet rearing. Particular reference is given to Chinese know-how and experience with Vespa rearing for medicinal and culinary motivations. We also draw on observations with reared insects that have a similar life history as Vespa hornets, namely Bombus bumblebees and Vespula yellowjackets. Key challenges to optimizing Vespa hornet rearing are identified and discussed.     

Tracing the invasion characteristics of the yellow-legged hornet,Vespa velutina nigrithorax (Hymenoptera: Vespidae), in Korea using newly detected variable mitochondrial DNA sequences

The yellow-legged hornet, Vespa velutina nigrithorax (Hymenoptera: Vespidae), invaded South Korea in 2003 through Busan metropolitan city, which is located in the southeast region of the country. Previous studies aiming to trace the origin of V. velutina in Korea used a portion of mitochondrial (mt) COI and detected a single haplotype common to the site of origin. However, no subsequent study on invasive dynamics such as additional entry and/or another site of entry has been performed. In this study, segments of mt COI, CytB, and lrRNA were sequenced from 238 individuals collected in 11 Korean and two Japanese localities, but no variation in each gene was observed. Thus, we developed two intergenic spacer (IGS) sequences from the publicly available mt genome of V. velutina, which provided substantially increased variability (i.e., 19 haplotypes with 1.74% maximum sequence divergence in 1,129–1,146-bp-long concatenated sequences). Population genetic analyses using the concatenated sequences unexpectedly provided higher genetic diversity estimates in the northwest and southwest regions, both of which also harbor international cargo ports, than in the southeast region, in which Busan is located. Furthermore, this genetic result was roughly concordant with our questionnaire survey demonstrating that V. velutina was observed in apiaries located in the northwest and southwest regions up to 2012, when there was no reported prevalent distribution of the hornet beyond the southeast region. These results collectively suggest that the northwest and southwest regions of Korea are additional sites of V. velutina entry to the country, independent from the southeast region origin.     

The invasion by the Yellow-legged hornet: A systematic review

The Yellow-legged hornet (Vespa velutina nigrithorax), native to regions of Southeast Asia, was accidentally introduced in Europe, South Korea, and Japan, where is has often become invasive. Due to its potential negative impacts at ecologic, economic and social levels, this hornet was included in the “Union list” of the EU legislation for invasive alien species. This means that measures are urgently needed to prevent further introductions, as well as to early-detect and control spread to avoid new populations. In this study we aim to identify the main reported drivers of distribution, ecological preferences, impacts, and methods for preventing introduction, controlling, and managing this invasive species. The supporting information was obtained from a comprehensive literature search. Then, a literature review was performed to classify the records gathered and to extract the relevant information following an adapted Drivers-Pressures-State-Impacts-Responses framework. The achieved results show a growing interest of researchers on V. velutina nigrithorax through time due to its quick spread and impacts on new ecosystems. They also revealed that there is much information on the State of invasions, whereas more knowledge is needed regarding the Drivers of those invasions. Biological traits such as life history traits, morphology, and the sting venom properties are some of the most studied topics regarding V. velutina nigrithorax. In the future, research should focus on the topics that lack information, analyse other Response solutions that meet the intended measures by the EU legislation, and use new methodology to study the impacts caused by this invader under new perspectives.     

Olfactory Attraction of the Hornet Vespa velutina to Honeybee Colony Odors and Pheromones

Since the beginning of the last century, the number of biological invasions has continuously increased worldwide. Due to their environmental and economical consequences, invasive species are now a major concern. Social wasps are particularly efficient invaders because of their distinctive biology and behavior. Among them, the yellow-legged hornet, Vespa velutina, is a keen hunter of domestic honeybees. Its recent introduction to Europe may induce important beekeeping, pollination, and biodiversity problems. Hornets use olfactory cues for the long-range detection of food sources, in this case the location of honeybee colonies, but the exact nature of these cues remains unknown. Here, we studied the orientation behavior of V. velutina workers towards a range of hive products and protein sources, as well as towards prominent chemical substances emitted by these food sources. In a multiple choice test performed under controlled laboratory conditions, we found that hornets are strongly attracted to the odor of some hive products, especially pollen and honey. When testing specific compounds, the honeybee aggregation pheromone, geraniol, proved highly attractive. Pheromones produced by honeybee larvae or by the queen were also of interest to hornet workers, albeit to a lesser extent. Our results indicate that V. velutina workers are selectively attracted towards olfactory cues from hives (stored food, brood, and queen), which may signal a high prey density. This study opens new perspectives for understanding hornets’ hunting behavior and paves the way for developing efficient trapping strategies against this invasive species.     

Review of the invasive yellow-legged hornet, <Emphasis Type="Italic">Vespa velutina nigrithorax</Emphasis> (Hymenoptera: Vespidae), in Japan and its possible chemical control

The distribution of the invasive yellow-legged hornet, Vespa velutina nigrithorax Buysson, is expanding in Europe and Asia. In Japan, this species was first detected on Tsushima Island, near Korea, in 2012. Because the rapid expansion of its distribution has raised public concern, the Japanese Ministry of the Environment added this hornet to the list of invasive alien species in 2015. In this paper, to contribute to the development of control techniques for this hornet, we present its phylogeny and life history. We then report its global expansion history and present status. Last, we review case studies of chemical control of vespine wasps and discuss effective techniques to control the hornet on Tsushima, where many Asian honeybee hives are kept. Successful case studies with Vespula wasps suggest the possibility of toxic baiting for the control of V. velutina nigrithorax.     

Competition between the native and the introduced hornets Vespa crabro and Vespa velutina: a comparison of potentially relevant life‐history traits

1. Invasive alien species are a major threat to biodiversity. In addition to predation and parasitism, native species might suffer from competition when invasive alien species occupy a similar ecological niche. 2. This study focused on the potential interspecific interaction between two hornets: the Asian yellow‐legged hornet, Vespa velutina, a high‐concern invasive alien species recently arrived in Europe; and the native European hornet, Vespa crabro. The two species share a similar ecological niche and V. velutina is rapidly expanding across Europe, which suggests that V. crabro might suffer from competition. 3. Under laboratory‐controlled conditions, two life‐history traits that might cause the two species to compete were investigated: (i) the ability of workers to find food sources and their flexibility in exploiting them (through individual food item choice tests and exploration assays); and (ii) the worker resistance to pathogens (through immune challenge tests). 4. The results show that trophic preference of both species highly overlaps, with a marked dietary preference for honeybees compared with other insect prey and non‐prey protein items. No differences were observed in the exploratory behaviour of both species. Finally, constitutive antibacterial activity was greater in workers of the native species than in workers of the invasive hornet. 5. This laboratory study provides a first assessment under controlled conditions of the factors affecting competition between workers of two hornet species and proposes a framework to assess, in wild contexts, the magnitude of the competition and the impact of the introduced V. velutina on the native V. crabro.     

Both olfactory and visual cues promote the hornet Vespa velutina to locate its honeybee prey Apis cerana

Predators use olfactory, visual and sometimes acoustic cues from the preys to assess food information. However, it is not known if the aggressive hornets (Vespa spp.) use olfactory, visual, or both types of information to find and recognize prey. In the present study, we trained hornet workers (Vespa velutina) to a feeding area. Once the hornets began consistently foraging at this feeding area, we determined whether they located prey (bees, Apis cerana) via olfactory or visual cues. We did this by testing whether hornets were attracted to a dummy bait (bee dummy bait or non-bee dummy bait) treated with extracts of honeybee cuticular hydrocarbons. We then tested whether hornets could distinguish between bee dummy bait and cotton ball dummy bait, both treated with bee odors. Hornets preferred the dummy treated with bee odors, and bee dummies (with bee images) were more attractive to the hornet than the cotton ball dummies with only bee odors. These results clearly indicate that a combination of olfactory and visual cues helps the hornet to locate its prey.     

Production of Early Diploid Males by European Colonies of the Invasive Hornet Vespa velutina nigrithorax

The invasive yellow-legged hornet Vespa velutina nigrithorax was accidentally introduced in Europe in the early 2000s. As is the case in colonies of other wasp and hornet species, V. velutina colonies are known to produce sexuals (males and new queens) at the end of the summer. We show that early-stage colonies in French populations frequently produce males well before the usual reproductive period. The vast majority of the males produced are diploid, which is consistent with the loss of genetic diversity previously reported in introduced populations in France. Since males do not participate in colony activities, the production of early diploid males at the expense of workers is expected to hamper colony growth and, ultimately, decrease the expansion of the species in its invasive range in Europe.     

Behavioral syndrome in a native and an invasive hymenoptera species

Recent studies have focused on the role of behavior in biological invasions. Individuals may differ consistently in time for several behavioral traits (personality) which covary (behavioral syndrome) resulting in different behavioral types, some of them favoring invasion. Social hymenopterans have a strong potential to be invaders and their success depends primarily on the foundresses’ ability to found viable colonies. They are expected to be active, explorative and bold for optimally establishing their nest. In Europe, 2 hornet species coexist: the native Vespa crabro and the invasive Vespa velutina. These 2 species may compete for nesting sites and we suggest that the initial success of V. velutina has been favored by its behavior in outperforming V. crabro for the traits involved in nest initiation. Here, we (i) defined the personality of V. crabro and V. velutina, (ii) tested for the existence of behavioral syndrome in these species, and (iii) compared their performances using an open‐field test. Our results show that V. crabro foundresses behave consistently but not V. velutina; this lack of consistency being mainly due to reduced variance among individuals. This result questions the possibility of detecting consistent behavioral differences in species having recently undergone a strong bottleneck. Both species exhibit the same correlations between activity, boldness and exploration and V. velutina clearly outperforms V. crabro for all traits. Our results suggest that activity, boldness, and exploration are implicated in both hornet nest initiation and invasion process which contributed to explain why social hymenopterans are so successful at colonization.     

Invasion by Solidago species has limited impacts on soil seed bank communities

Increasing attention in invasion biology is being paid to measuring and understanding the impacts of invasive species. For plant invasions, however, the impact of invasion on soil seed bank communities has been under-studied. At six sites in southern Germany, we investigated whether areas invaded by Solidago gigantea and Solidago canadensis experienced a reduction in seed bank species richness, size and diversity, and a change in species composition compared to adjacent uninvaded areas. We found no overall effect of invasion on seed bank size, or on species richness and diversity. Seed bank size significantly decreased from 0–5 cm to 5–10 cm depth in both invaded and uninvaded areas. A significant amount of variation in species composition was explained by invasion, but it was only one-tenth of that explained solely by site effects. Our study suggests that invasion by Solidago species may not have the same impacts on the soil seed banks of native species as other invasive perennial forbs that have so far been studied.     

Genetic diversity of genus Vespa including an invaded species of V. velutina (Hymenoptera: Vespidae) in Korea inferred from DNA barcoding data

With about 5000 known species, the Vespidae is a large family belongs to order Hymenoptera. The genus Vespa with 22 species is one of the four genera of the subfamily Vespinae. In Korea, 10 species and subspecies are recognized. Because of their social behavior, their treat to human health and their impact in apiculture, the reliable and sometimes automated identification of these insects to species level are important. To test the efficacy of DNA barcoding method for identification of species of the genus Vespa in Korea, 30 samples of eight Korean species of genus Vespa were collected and mitochondrial DNAs of 658 bp fragment cytochrome oxidase subunit 1 (CO1) region were sequenced. A Bayesian Inference based on COI gene of the Korean Vespa species was constructed. The phylogenetic tree shoed that identification of all specimens is possible based on COI gene and we found strong relation between the sequences of the collected species from different localities in South Korea which clustered together with 100% support with sequences of the same species in GenBank. The results demonstrate that DNA barcoding is a useful technique for rapid and accurate species recognition in Korean Vespa species. The DNA barcode part of COI for V. binghami is provided for the first time that can help for identification of this species through DNA barcoding. Also, the genetic diversity among Korean Vespa velutina was zero suggests that the invasion might have occurred in a single event with small number of founders.     

Does salt stress increase the ability of the exotic legume Acacia longifolia to compete with native legumes in sand dune ecosystems?

Sand dune ecosystems are one of the areas most affected by the introduction of invasive species which represents a threat for biodiversity conservation. Their invasion patterns and spread may depend on their salinity tolerance, besides other factors. To test this hypothesis, we investigated the effects of salt stress on seed germination and on the activity of antioxidant enzymes (catalase, CAT; ascorbate peroxidase, APX; peroxidase, POX; and glutathione reductase, GR) in two legume species, an invasive, Acacia longifolia (Andrews.) Willd., and a native, Ulex europaeus (L.), very common in the sand dunes of the coast of Portugal. Salt stress was induced by adding NaCl at different concentrations, 0, 50, 100 and 200 mM, for 15 days. Results showed that the highest germination percentages were obtained in distilled water (control) and that, with increasing salt concentration, seed germination was delayed and decreased in both species. Inhibition of germination was higher in the native species, only 3% of seeds germinated at 100 mM and no seeds germinated at 200 mM NaCl. In the invasive species, the reduction was higher at 200 mM NaCl (16%). Considering the coefficient of germination velocity, a decrease in both species with increasing NaCl concentration was observed. The CAT and GR activities decreased in A. longifolia with increasing salinity. In turn, APX activity significantly increased as NaCl concentration increased while the POX activities declined at the highest NaCl concentration. On the other hand, at 50 mM NaCl lower activity of CAT and APX and higher GR and POX were found in U. europaeus. In both species, protein content increased as NaCl concentration increased. In addition, it seems that APX activities play an essential role in the scavenging reactive oxygen species (ROS). These results suggest that the seeds of the invasive legume A. longifolia are more tolerant to salinity than the native legume U. europaeus, and seem better equipped to handle the physiological stress of high salinity, which may contribute to its invasive ability in sand dunes.     

First detection of <Emphasis Type="Italic">Vespa velutina nigrithorax</Emphasis> (Hymenoptera: Vespidae) in the Balearic Islands (Western Mediterranean): a challenging study case

The yellow-legged hornet, Vespa velutina nigrithorax is an Invasive Alien Species introduced into Europe in 2004, and is currently present in France, Italy, Spain, Portugal, Germany, and Belgium. It was also recently introduced in the United Kingdom and Switzerland. Here we present the first detection of V. velutina in the westernmost Mediterranean islands (Majorca, Balearic archipelago). In October 2015, a specimen of hornet, which was later identified as V. velutina, was sent to the laboratory of Zoology at the University of the Balearic Islands by a beekeeper from the locality of Sóller (northwest of Majorca). After the identification, and in coordination with the local authorities, an intensive survey was implemented in the area of first detection using sticky traps baited with protein and nest detection by directly observing the flight route of adult hornets from two or three feeding points. The first nest was detected on 30th October 2015 and was entirely removed. The next year, nine other nests were located from August to November 2016 in the same area and again, all of them were removed entirely. It is the first report on an island of destroying nests as a means of controlling the spread, a scenario very different to mainland Europe. All nests were detected in evergreen tree species (pines, holm oaks, and common cypress) in the “Serra de Tramuntana”, a mountain range located in the north-west of the island of Majorca (3667 km²), which is situated 176 km off the mainland. Considering the presumably early stage of the invasion of V. velutina in Majorca, the size of the island, and the confinement of the detection area, we consider that this species could be eradicated if resources and a specific program for eradication in Majorca are implemented in coordination with all involved partners.     

How <Emphasis Type="Italic">Apis mellifera</Emphasis> Behaves with its Invasive Hornet Predator <Emphasis Type="Italic">Vespa velutina</Emphasis>?

Invasive species are now recognized as a major cause of native biodiversity loss worldwide. In the current deleterious context for pollinators, the invasive yellow-legged hornet, Vespa velutina, represents an additional threat to the domestic honeybee, Apis mellifera, in Europe. Therefore, understanding the impact of this predator on honeybee colonies is of major importance. In the present study, we tried to assess the impact of V. velutina on the honeybee foraging and defence behaviour based on the video monitoring of two hives. Balling behaviour is reported here for the first time under natural conditions in A. mellifera against V. velutina in Europe. Although these results are preliminary and should be carefully considered, we found that the number of hornets impacted honeybee foraging and defence behaviours. More defensive behaviours were notified in the hive, which survives slightly longer. This may suggest that selecting for more defensive colonies may provide an interesting perspective.     

Vespa velutina nest distribution at a local scale: An 8‐year survey of the invasive honeybee predator

Vespa velutina is an invasive species that was observed for the first time in France and Europe in 2004, which rapidly threatened domestic honeybees with active predation. For the first time in this study, we analyzed the distribution of V. velutina nests at a local scale to understand the pattern of nesting, and in 8 years of monitoring, a total of 528 nests were surveyed. With the exception of 2 years, the nests of V. velutina were randomly distributed within the monitored area, which suggested that intraspecific competition for nesting sites and/or food was unlikely. When the data for all years were combined, an aggregated distribution of nests at the seafront in the vicinity of the oyster farm and sportfishing activities was observed. This distribution suggested that seafood, attractive to vespids, might provide a valuable alternative food source that favored colony development. We also tested the effect of spring queen trapping of V. velutina on the distribution of nests in 2014, which was the year with the greatest number of colonies within the area; the position of the traps did not influence the distribution of the nests. For the first time, this study showed that (i) intraspecific competition was unlikely to be a mechanism for population regulation, (ii) some areas were more likely favored for hornet nests, and (iii) spring queen wasp trapping was inefficient as a method to limit the distribution of the V. velutina population.     

Variable-temperature 13C solid-state NMR study of the molecular structure of honeybee wax and silk

To elucidate the native-state crystal structure of beeswax from the Japanese bee, Apis cerana japonica, we determined the relationship between temperature and the ¹³C solid-state nuclear magnetic resonance (NMR) chemical shift of methylene carbon of beeswax, with comparison to n-alkanes and polyethylene in the orthorhombic, monoclinic, or triclinic crystal form. Variable-temperature ¹³C solid-state NMR observations of n-alkanes and polyethylene revealed that the chemical shifts of methylene carbon in the orthorhombic crystal form increased linearly with increasing temperature, that of the triclinic form decreased, and that of the monoclinic form was unaltered. These relations were compared with results of variable-temperature ¹³C solid-state NMR observation of beeswax. Results clarified that the two crystal forms comprising the beeswax in the native state are orthorhombic and monoclinic. The variable-temperature ¹³C solid-state NMR observations were also applied to interpret the differential scanning calorimetry (DSC) curve of beeswax. They were used to clarify the structural changes of beeswax for widely various temperatures. For beeswax secreted by the Japanese bee, the transition from the orthorhombic form to the rotator phase occurred at 36 °C, that is from the crystalline to the intermediate state at 45 °C. Moreover, the variable-temperature ¹³C solid-state NMR spectrum of honeybee silk in the native state was observed. Results demonstrated that the secondary structures of honeybee silk proteins in the native state comprised coexisting α-helix and β-sheet conformations and that the amount of α-helices was greater. The α-helix content of honeybee silk was compared with that of hornet silk produced by Vespa larvae.     

Community structure of ground beetles (Coleoptera: Carabidae) along an altitudinal gradient on Mt. Sobaeksan,Korea

We conducted this study to determine the diversity patterns, community structures, and seasonality of ground beetle assemblages along an altitudinal gradient (437–1420 m) on Mt. Sobaeksan, Korea. Ground beetles were collected by pitfall traps installed along an altitudinal gradient (437, 757, 1100, and 1420 m). A total of 32 species belonging to seven subfamilies were identified from 3259 collected ground beetles. The diversity pattern of the ground beetle assemblage according to altitude was neither monotonic decreased nor hump-shaped. However, subfamily assemblages and wing form diversity patterns differed according to altitude and may be correlated with altitude or some other environmental variables. The dominant species were Synuchus cycloderus (29.4%) and Eucarabus sternbergi sobaeksanensis (15.4%) and their seasonal activities according to altitude were similar. According to non-metric multidimensional scaling, ground beetles and altitudes could be divided into two distinct groups: a low altitude group (437–757 m) and a high altitude group (1100–1420 m). Some species were particularly abundant at high altitudes, such as Aulonocarabus koreanus kwonileeique and Poecilus nitidicollis, while others were highly abundant at low altitudes, such as Pterostichus audax, Pterostichus ishikawai, and Synuchus species.     

Options for the biological and physical control of Vespa velutina nigrithorax (Hym.: Vespidae) in Europe: A review

Recently, the environmental economist J.M. Salles (Salles, 2016 ) declared that “the Asian hornet was likely to be the most threatening invader insect in France.” Thirteen years after the accidental introduction in France, the Asian hornet (Vespa velutina nigrithorax) has invaded most west European countries. Little has been done to date to limit its progression and its economic, ecological and social impact. Although it is illusory to try to eradicate this species, it is known that a targeted control would limit its threatening trend. Current V. velutina control in France is mainly based on (i) high scale volunteer trapping by citizens and (ii) volunteer spotting of the nest. Evaluation of trapping strategies developed so far to control V. velutina expansion has highlighted their failure and has demonstrated the need to optimize nest detection techniques and to investigate on new control strategies. This review describes most of the means aimed to control predation and expansion of V. velutina, whether they have been scientifically assessed or only tested on field with decent success. Published prospective control methods and biological control techniques are also presented.     

Life cycle of Anthracophora rusticola (Coleoptera: Cetoniidae) from the commensal nests of Chinese Sparrowhawks (Accipiter soloensis)

Commensal breeding habits of the Anthracophora rusticola (Coleoptera: Cetoniidae) in stick nests of carnivorous, piscivorous, and omnivorous birds were recently reported, but the detailed life history of the scarab beetle has not been thoroughly elucidated. To understand the breeding cycle of the declining A. rusticola related to that of nesting Chinese sparrowhawks (Accipiter soloensis) in Korea, we examined six sparrowhawk nests from 2005 to 2008 and reared larvae and cocoons in a cage in 2008. Each sparrowhawk nest hosted 11.3 ± 3.0 larvae and cocoons, implying that sparrowhawk nests might be the most important nesting resource of the beetles in Korea. The beetles may lay their eggs between late May and early July after nest building by sparrowhawks. Larvae were observed from July to August in the sparrowhawk nests, and adult beetles emerged in September after a mean pupal period of 25.5 days. Our results on the coinciding breeding of two commensal taxa over three months from May to September may provide key information regarding the life history of the commensal scarabid beetle and potential causes of the concurrent decline of both taxa in Korea.