Diversity and distribution of orchid bees (Hymenoptera: Apidae, Euglossini) in Belize.

Background: Orchid bees are abundant and widespread in the Neotropics, where males are important pollinators of orchids they visit to collect fragrant chemicals later used to court females. Assemblages of orchid bees have been intensively surveyed in parts of Central America, but less so in Belize, where we studied them during the late-wet and early-dry seasons of 2015-2020. Methods: Using bottle-traps baited with chemicals known to attract a variety of orchid bee species, we conducted surveys at sites varying in latitude, historical annual precipitation, elevation, and the presence of nearby agricultural activities. Each sample during each survey period consisted of the same number of traps and the same set of chemical baits, their positions randomized along transects. Results: In 86 samples, we collected 24 species in four genera: Euglossa (16 species), Eulaema (3), Eufriesea (3), and Exaerete (2). During our most extensive sampling (December 2016-February 2017), species diversity was not correlated with latitude, precipitation, or elevation; species richness was correlated only with precipitation (positively). However, a canonical correspondence analysis indicated that species composition of assemblages varied across all three environmental gradients, with species like Eufriesea concava, Euglossa imperialis, and Euglossa viridissima most common in the drier north, and Euglossa ignita, Euglossa purpurea, and Eulaema meriana more so in the wetter southeast. Other species, such as Euglossa tridentata and Eulaema cingulata, were common throughout the area sampled. Mean species diversity was higher at sites with agricultural activities than at sites separated from agricultural areas. A Chao1 analysis suggests that other species should yet be found at our sites, a conclusion supported by records from adjacent countries, as well as the fact that we often added new species with repeated surveys of the same sites up through early 2020, and with the use of alternative baits. Additional species may be especially likely if sampling occurs outside of the months/seasons that we have sampled so far.

Review of Paraclius arcuatus species-group (Diptera, Dolichopodidae) with description of two new species.

The Paraclius arcuatus species-group is reviewed and includes the following seven species: P. angusticauda Van Duzee, 1933, P. angustipennis Van Duzee, 1929, P. arcuatus (Loew, 1861), P. brooksi sp. nov., P. elongatus Van Duzee, 1930, P. pumilio Loew, 1872, and P. xibun sp. nov. Paraclius angustipennis, P. arcuatus and P. pumilio are redescribed based on photographs of types and additional specimens. Paraclius femoratus Aldrich, 1901 is synonymized with P. arcuatus, syn. nov. and Paraclius magnicornis Van Duzee, 1927 is regarded as a junior synonym of P. pumilio, syn. nov. An identification key, distribution maps and high-resolution photographs of species are provided. In addition, the systematic position of Paraclius is discussed.

Constitutive and Induced Defenses in Long-lived Pines Do Not Trade Off but Are Influenced by Climate.

Plants resist herbivores and pathogens by using constitutive (baseline) and inducible (change in defense after an attack) defenses. Inducibility has long been predicted to trade off with constitutive defense, reflecting the economic use of resources. However, empirical evidence for such tradeoffs is variable, and we still lack understanding about when and where defense trade-offs occur. We tested for tradeoffs between constitutive and induced defenses in natural populations of three species of long-lived pines (Pinus balfouriana, P. flexilis, P. longaeva) that differ greatly in constitutive defense and resistance to mountain pine beetle (MPB, Dendroctonus ponderosae). We also assessed how climate influenced constitutive and inducible defenses. At seven high-elevation sites in the western U.S., we simulated MPB attack to induce defenses and measured concentrations of terpene-based phloem defenses on days 0, 15, and 30. Constitutive and induced defenses did not trade off among or within species. Simulated MPB attack induced large increases in defense concentrations in all species independent of constitutive levels. MPB and its symbiotic fungi typically kill trees and thus could be selective forces maintaining strong inducibility within and among species. The contrasting constitutive concentrations in these species could be driven by the adaptation for specializing in harsh, high-elevation environments (e.g., P. balfouriana and P. longaeva) or by competition (e.g., P. flexilis), though these hypotheses have not been empirically examined. Climate influenced defenses, with the greatest concentrations of constitutive and induced defenses occurring at the coldest and driest sites. The interactions between climate and defenses have implications for these species under climate change.

Nematode-induced demasculinization of Nearctic Dolichopodidae (Diptera) with five new synonyms.

Demasculinization, the reduction of male sexual characters due to parasitic nematode infection, is described in Nearctic Dolichopodidae. Five species are confirmed to be demasculinized (3 species of Dolichopus Latreille and 1 species each of Rhaphium Meigen and Tachytrechus Haliday) by nematodes in two families (apparent Mermithidae and Parasitylenchidae). The following nematode-infected males were inadvertently described as new species and are here proposed as synonyms: Dolichopus hurleyi Runyon, 2008 = D. dorycerus Loew, 1864, syn. nov.; Dolichopus frosti Runyon, 2008 = D. sincerus Melander, 1900, syn. nov.; Tachytrechus boharti Harmston, 1968 = T. sanus Osten Sacken, 1877, syn. nov.; Tachytrechus duplicatus Harmston, 1972 = T. sanus Osten Sacken, 1877, syn. nov.; Tachytrechus mchughi Harmston, 1972 = T. sanus Osten Sacken, 1877, syn. nov. A conspectus is presented of the scope of taxonomic problems caused by nematodes in Nearctic Dolichopus and Tachytrechus, which suggests that ten additional names in Dolichopus could be based on infected specimens. Guidelines are provided for recognizing nematode infection to avoid erroneous species descriptions.

A new species of Chrysotus Meigen (Diptera, Dolichopodidae) from soybean fields in South Dakota, USA.

A new long-legged fly species, Chrysotussoya sp. nov. (Diptera: Dolichopodidae), is described and illustrated from specimens collected in soybean fields near Brookings, South Dakota, USA. The abundance of this species in soybeans suggests it plays an important role as a beneficial predator.

Wild Bee Response to Application of the Douglas-fir Beetle Anti-Aggregation Pheromone, 3-Methylcyclohex-2-En-1-One.

Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (DFB), is the most damaging insect pest of Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, in western North America. Individual high-value trees and stands can be protected during DFB outbreaks using the beetle's anti-aggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH), which is available in several commercial formulations. However, other methylcyclohexanones similar in molecular structure to MCH have been shown to repel bees from agricultural areas. In forest systems, wild bees (Hymenoptera: Apiformes) provide essential pollination services to many forb and shrub species. Therefore, it is important to determine whether resident wild bee communities are affected in forests treated with MCH. To address this, the bee community was sampled within experimental sites located in Idaho and Montana, USA in 2016. At both sites, there was no significant difference in overall bee abundance, species richness, or diversity between MCH-treated plots and untreated control plots. Overall, these results indicate that treatment of Douglas-fir with MCH does not negatively impact the resident wild bee community.

Parasitylenchus myiophagus n. sp. (Nematoda: Parasitylenchidae), a tylenchid nematode parasite of long-legged flies (Diptera: Dolichopodidae).

A tylenchid nematode parasite of a male long-legged fly, Tachytrechus sanus Osten Sacken (Diptera: Dolichopodidae) from Montana, USA is described as Parasitylenchus myiophagus n. sp. (Nematoda: Parasitylenchidae). The new species is characterized by the presence of an extremely long first generation female and numerous short and wide second generation males and females produced in enclosed clusters in the hosts body cavity. Both generation female nematodes are ovoviviparous, with short stylets lacking knobs and simple tails lacking spikes, palps or mucrons. The second generation males have paired, separate spicules, short stylets, and a bursa but no visible gubernaculum. The fly host shows evidence of demasculinization, which is attributed to nematode parasitism. The gonads of the second generation adults are infected with a microsporidium (Microsporidia), which is a new host record for tylenchid nematodes. A tylenchid-infected Baltic amber dolichopodid shows that associations between these two organisms extend back at least to the Eocene.

Changes in the Summer Wild Bee Community Following a Bark Beetle Outbreak in a Douglas-fir Forest.

The status of wild bees has received increased interest following recent estimates of large-scale declines in their abundances across the United States. However, basic information is limited regarding the factors affecting wild bee communities in temperate coniferous forest ecosystems. To assess the early responses of bees to bark beetle disturbance, we sampled the bee community of a Douglas-fir, Pseudotsuga menziesii (Mirb.), forest in western Idaho, United States during a Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (Coleoptera: Curculionidae), outbreak beginning in summer 2016. We resampled the area in summer 2018 following reductions in forest canopy cover resulting from mortality of dominant and codominant Douglas-fir. Overall, results from rarefaction analyses indicated significant increases in bee diversity (Shannon's H) in 2018 compared to 2016. Results from ANOVA also showed significant increases in bee abundance and diversity in 2018 compared to 2016. Poisson regression analyses revealed percent tree mortality from Douglas-fir beetle was positively correlated with increases in total bee abundance and species richness, where community response variables displayed a cubic trend with percent tree mortality. Percent reduction in canopy cover from 2016 to 2018 was also correlated with bee species richness and diversity. These findings suggest that wild bee communities may benefit from changes in forest structure following bark beetle outbreaks.

The Dolichopodidae (Diptera) of Montserrat, West Indies.

The long-legged flies (Dolichopodidae) of the island of Montserrat in the Lesser Antilles have been surveyed and include 63 species in 27 genera. The following eleven new species are described and illustrated: Amblypsilopus marskeae sp. nov., Medetera iviei sp. nov., Medetera montserratensis sp. nov., Systenus ladonnae sp. nov., Thrypticus mediofuscus sp. nov., Chrysotus antillensis sp. nov., Chrysotus callichromoides sp. nov., Chrysotus interfrons sp. nov., Chrysotus montserratensis sp. nov., Diaphorus robinsoni sp. nov., and Sympycnus montserratensis sp. nov. Six species have only been found on Montserrat (ca. 10% endemicity). Keys are provided to the genera and species on Montserrat, their known distribution summarized, and additional new island records provided for many species. Asyndetus wirthi Robinson is synonymized with A. interruptus (Loew) and Achradocera apicalis (Aldrich) is removed from synonymy with A. barbata (Loew). Diaphorus flavipes Aldrich is transferred to Chrysotus as a new combination. A new replacement name, Chrysotus milvadu nom. nov., is provided for the Nearctic Chrysotus parvulus Van Duzee. Lectotypes are designated for Achradocera apicalis (Aldrich) and Asyndetus fratellus Aldrich. The fauna of Montserrat is summarized and compared with that of Dominica. Collecting methods are compared and threats to the dolichopodid fauna of Montserrat discussed.

A Biodegradable Formulation of MCH (3-Methylcyclohex-2-en-1-one) for Protecting Pseudotsuga menziesii from Dendroctonus pseudotsugae (Coleoptera: Curculionidae) Colonization.

Douglas-fir, Pseudotsuga menziesii (Mirb.) Franco, trees and stands can be protected from Douglas-fir beetle, Dendroctonus pseudotsugae Hopkins (DFB)-caused mortality by application of synthetic formulations of the beetle's antiaggregation pheromone, 3-methylcyclohex-2-en-1-one (MCH). A biodegradable formulation of MCH, SPLAT MCH, was developed and evaluated for protecting individual Douglas-fir trees and small stands from colonization and mortality by DFB. In an individual-tree experiment in Idaho, both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of treated trees colonized and killed by DFB compared to untreated controls. SPLAT MCH was as effective as MCH bubble capsules for protecting individual trees. Both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of trees colonized and killed by DFB within 0.04-ha circular plots surrounding each treated tree compared to untreated controls. In 0.41 ha stands in New Mexico, both MCH bubble capsules and SPLAT MCH significantly reduced the proportion of trees colonized and killed by DFB compared to untreated controls, again with no differences observed between MCH treatments. In a similar stand level trial in Idaho, neither MCH treatment significantly reduced the proportion of trees colonized by DFB, and only MCH bubble capsules significantly reduced levels of tree mortality compared to untreated controls, but no significant difference was observed between SPLAT MCH and MCH bubble capsules. Overall, the results indicate that SPLAT MCH is as effective as MCH bubble capsules for protecting individual trees and small stands of Douglas-fir from DFB-caused mortality.

Volatiles of High-Elevation Five-Needle Pines: Chemical Signatures through Ratios and Insight into Insect and Pathogen Resistance.

High-elevation five-needle pine trees are a group of Pinus species in the subgenus Strobus that occur at the edges of plant growth near the alpine tree line. These species are ecologically very important and are also threatened by climate-driven insect outbreaks and an exotic pathogen. Volatile organic compounds (VOCs) play central roles in the environmental adaptation of plants and in their defense against insects and pathogens. For example, the VOCs emitted by some high-elevation five-needle pine species attract female, tree-killing mountain pine beetles (MPB, Dendroctonus ponderosae) in the pioneering phase whereas VOCs from other species strongly repel this foremost herbivore, but the mechanism is unknown. We collected and compared headspace VOCs from foliage of eight species of high-elevation five-needle pines in Europe and North America. Overall, VOCs differed quantitatively among species with few qualitative differences. Despite species emitting essentially the same compounds, Random Forest analysis correctly classified 117 of the 126 trees sampled by using VOCs and identified the most important compounds for species classification and for separating species resistant from those susceptible to MPB or white pine blister rust (Cronartium ribicola). These VOC 'fingerprints' resulted largely from species emitting distinctive ratios of compounds, rather than through presence of species-specific compounds. Importantly, these Pinus species vary greatly in resistance to the main herbivore (MPB) and pathogen (white pine blister rust) causing tree mortality. Thus, these findings provide insights and should guide research into understanding resistance and in developing tools to manage these important trees. For instance, studies into the functions of five-needle pine VOCs in defense against abiotic or biotic stressors should focus on blend ratios rather than on individual compounds.

Two new species of Hurleyella Runyon amp; Robinson (Diptera: Dolichopodidae), with the first record from the Neotropics.

Two new species of the long-legged fly genus Hurleyella Runyon Robinson, 2010 are described and illustrated: Hurleyella belizensis sp. nov. from Belize and Hurleyella salina sp. nov. from alkali areas of the Northern Rockies of the USA (Idaho, Montana, Wyoming). The discovery of these new species greatly extends the known distribution of Hurleyella northward in the Nearctic and southward into the Neotropics. Notes, photos of habitats, a distribution map, and a key to the four known species of Hurleyella are provided.

Comprehensive inventory of true flies (Diptera) at a tropical site.

Estimations of tropical insect diversity generally suffer from lack of known groups or faunas against which extrapolations can be made, and have seriously underestimated the diversity of some taxa. Here we report the intensive inventory of a four-hectare tropical cloud forest in Costa Rica for one year, which yielded 4332 species of Diptera, providing the first verifiable basis for diversity of a major group of insects at a single site in the tropics. In total 73 families were present, all of which were studied to the species level, providing potentially complete coverage of all families of the order likely to be present at the site. Even so, extrapolations based on our data indicate that with further sampling, the actual total for the site could be closer to 8000 species. Efforts to completely sample a site, although resource-intensive and time-consuming, are needed to better ground estimations of world biodiversity based on limited sampling.

Remarkable fly (Diptera) diversity in a patch of Costa Rican cloud forest: Why inventory is a vital science.

Study of all flies (Diptera) collected for one year from a four-hectare (150 x 266 meter) patch of cloud forest at 1,600 meters above sea level at Zurquí de Moravia, San José Province, Costa Rica (hereafter referred to as Zurquí), revealed an astounding 4,332 species. This amounts to more than half the number of named species of flies for all of Central America. Specimens were collected with two Malaise traps running continuously and with a wide array of supplementary collecting methods for three days of each month. All morphospecies from all 73 families recorded were fully curated by technicians before submission to an international team of 59 taxonomic experts for identification.        Overall, a Malaise trap on the forest edge captured 1,988 species or 51% of all collected dipteran taxa (other than of Phoridae, subsampled only from this and one other Malaise trap). A Malaise trap in the forest sampled 906 species. Of other sampling methods, the combination of four other Malaise traps and an intercept trap, aerial/hand collecting, 10 emergence traps, and four CDC light traps added the greatest number of species to our inventory. This complement of sampling methods was an effective combination for retrieving substantial numbers of species of Diptera. Comparison of select sampling methods (considering 3,487 species of non-phorid Diptera) provided further details regarding how many species were sampled by various methods.        Comparison of species numbers from each of two permanent Malaise traps from Zurquí with those of single Malaise traps at each of Tapantí and Las Alturas, 40 and 180 km distant from Zurquí respectively, suggested significant species turnover. Comparison of the greater number of species collected in all traps from Zurquí did not markedly change the degree of similarity between the three sites, although the actual number of species shared did increase.        Comparisons of the total number of named and unnamed species of Diptera from four hectares at Zurquí is equivalent to 51% of all flies named from Central America, greater than all the named fly fauna of Colombia, equivalent to 14% of named Neotropical species and equal to about 2.7% of all named Diptera worldwide. Clearly the number of species of Diptera in tropical regions has been severely underestimated and the actual number may surpass the number of species of Coleoptera.        Various published extrapolations from limited data to estimate total numbers of species of larger taxonomic categories (e.g., Hexapoda, Arthropoda, Eukaryota, etc.) are highly questionable, and certainly will remain uncertain until we have more exhaustive surveys of all and diverse taxa (like Diptera) from multiple tropical sites.        Morphological characterization of species in inventories provides identifications placed in the context of taxonomy, phylogeny, form, and ecology. DNA barcoding species is a valuable tool to estimate species numbers but used alone fails to provide a broader context for the species identified.

Palpi aplenty: new species in the Chrysotus longipalpus species group (Diptera: Dolichopodidae).

Four new Nearctic species belonging to the Chrysotus longipalpus species group are described: Chrysotus keyensis sp. nov. (Florida), Chrysotus mccreadiei sp. nov. (Alabama), Chrysotus mystax sp. nov. (Alabama), and Chrysotus plumarista sp. nov. (Alabama). This brings the number of known species in this group to twelve. A key to species of males of the C. longipalpus species group and new distribution records are provided.

Drought and increased CO2 alter floral visual and olfactory traits with context-dependent effects on pollinator visitation.

Climate change can alter species interactions essential for maintaining biodiversity and ecosystem function, such as pollination. Understanding the interactive effects of multiple abiotic conditions on floral traits and pollinator visitation are important to anticipate the implications of climate change on pollinator services. Floral visual and olfactory traits were measured from individuals of four forb species subjected to drought or normal water availability, and elevated or ambient concentrations of CO2 in a factorial design. Pollinator visitation rates and community composition were observed in single-species and multi-species forb assemblages. Drought decreased floral visual traits and pollinator visitation rates but increased volatile organic compound (VOC) emissions, whereas elevated CO2 positively affected floral visual traits, VOC emissions and pollinator visitation rates. There was little evidence of interactive effects of drought and CO2 on floral traits and pollinator visitation. Interestingly, the effects of climate treatments on pollinator visitation depended on whether plants were in single- or multi-species assemblages. Components of climate change altered floral traits and pollinator visitation, but effects were modulated by plant community context. Investigating the response of floral traits, including VOCs, and context-dependency of pollinator attraction provides additional insights and may aid in understanding the overall effects of climate change on plant-pollinator interactions.

Global change effects on plant-insect interactions: the role of phytochemistry.

Natural and managed ecosystems are undergoing rapid environmental change due to a growing human population and associated increases in industrial and agricultural activity. Global environmental change directly and indirectly impacts insect herbivores and pollinators. In this review, we highlight recent research examining how environmental change factors affect plant chemistry and, in turn, ecological interactions among plants, herbivores, and pollinators. Recent studies reveal the complex nature of understanding global change effects on plant secondary metabolites and plant-insect interactions. Nonetheless, these studies indicate that phytochemistry mediates insect responses to environmental change. Future research on the chemical ecology of plant-insect interactions will provide critical insight into the ecological effects of climate change and other anthropogenic disturbances. We recommend greater attention to investigations examining interactive effects of multiple environmental change factors in addition to chemically mediated plant-pollinator interactions, given limited research in these areas.

The smell of environmental change: Using floral scent to explain shifts in pollinator attraction.

As diverse environmental changes continue to influence the structure and function of plant-pollinator interactions across spatial and temporal scales, we will need to enlist numerous approaches to understand these changes. Quantitative examination of floral volatile organic compounds (VOCs) is one approach that is gaining popularity, and recent work suggests that floral VOCs hold substantial promise for better understanding and predicting the effects of environmental change on plant-pollinator interactions. Until recently, few ecologists were employing chemical approaches to investigate mechanisms by which components of environmental change may disrupt these essential mutualisms. In an attempt to make these approaches more accessible, we summarize the main field, laboratory, and statistical methods involved in capturing, quantifying, and analyzing floral VOCs in the context of changing environments. We also highlight some outstanding questions that we consider to be highly relevant to making progress in this field.

Signals of speciation: volatile organic compounds resolve closely related sagebrush taxa, suggesting their importance in evolution.

Volatile organic compounds (VOCs) play important roles in the environmental adaptation and fitness of plants. Comparison of the qualitative and quantitative differences in VOCs among closely related taxa and assessing the effects of environment on their emissions are important steps to deducing VOC function and evolutionary importance. Headspace VOCs from five taxa of sagebrush (Artemisia, subgenus Tridentatae) growing in two common gardens were collected and analyzed using GC-MS. Of the 74 total VOCs emitted, only 15 were needed to segregate sagebrush taxa using Random Forest analysis with a low error of 4%. All but one of these 15 VOCs showed qualitative differences among taxa. Ordination of results showed strong clustering that reflects taxonomic classification. Random Forest identified five VOCs that classify based on environment (2% error), which do not overlap with the 15 VOCs that segregated taxa. We show that VOCs can discriminate closely related species and subspecies of Artemisia, which are difficult to define using molecular markers or morphology. Thus, it appears that changes in VOCs either lead the way or follow closely behind speciation in this group. Future research should explore the functions of VOCs, which could provide further insights into the evolution of sagebrushes.

Drought and leaf herbivory influence floral volatiles and pollinator attraction.

The effects of climate change on species interactions are poorly understood. Investigating the mechanisms by which species interactions may shift under altered environmental conditions will help form a more predictive understanding of such shifts. In particular, components of climate change have the potential to strongly influence floral volatile organic compounds (VOCs) and, in turn, plant-pollinator interactions. In this study, we experimentally manipulated drought and herbivory for four forb species to determine effects of these treatments and their interactions on (1) visual plant traits traditionally associated with pollinator attraction, (2) floral VOCs, and (3) the visitation rates and community composition of pollinators. For all forbs tested, experimental drought universally reduced flower size and floral display, but there were species-specific effects of drought on volatile emissions per flower, the composition of compounds produced, and subsequent pollinator visitation rates. Moreover, the community of pollinating visitors was influenced by drought across forb species (i.e. some pollinator species were deterred by drought while others were attracted). Together, these results indicate that VOCs may provide more nuanced information to potential floral visitors and may be relatively more important than visual traits for pollinator attraction, particularly under shifting environmental conditions.