FAVIS: Fast and versatile protocol for non-destructive metabarcoding of bulk insect samples.

Insects are diverse and sustain essential ecosystem functions, yet remain understudied. Recent reports about declines in insect abundance and diversity have highlighted a pressing need for comprehensive large-scale monitoring. Metabarcoding (high-throughput bulk sequencing of marker gene amplicons) offers a cost-effective and relatively fast method for characterizing insect community samples. However, the methodology applied varies greatly among studies, thus complicating the design of large-scale and repeatable monitoring schemes. Here we describe a non-destructive metabarcoding protocol that is optimized for high-throughput processing of Malaise trap samples and other bulk insect samples. The protocol details the process from obtaining bulk samples up to submitting libraries for sequencing. It is divided into four sections: 1) Laboratory workspace preparation; 2) Sample processing-decanting ethanol, measuring the wet-weight biomass and the concentration of the preservative ethanol, performing non-destructive lysis and preserving the insect material for future work; 3) DNA extraction and purification; and 4) Library preparation and sequencing. The protocol relies on readily available reagents and materials. For steps that require expensive infrastructure, such as the DNA purification robots, we suggest alternative low-cost solutions. The use of this protocol yields a comprehensive assessment of the number of species present in a given sample, their relative read abundances and the overall insect biomass. To date, we have successfully applied the protocol to more than 7000 Malaise trap samples obtained from Sweden and Madagascar. We demonstrate the data yield from the protocol using a small subset of these samples.

Temperature accounts for the biodiversity of a hyperdiverse group of insects in urban Los Angeles.

The urban heat island effect is a worldwide phenomenon that has been linked to species distributions and abundances in cities. However, effects of urban heat on biotic communities are nearly impossible to disentangle from effects of land cover in most cases because hotter urban sites also have less vegetation and more impervious surfaces than cooler sites within cities. We sampled phorid flies, one of the largest, most biologically diverse families of true flies (Insecta: Diptera: Phoridae), at 30 sites distributed within the central Los Angeles Basin, where we found that temperature and the density of urban land cover are decoupled. Abundance, richness, and community composition of phorids inside urban Los Angeles were most parsimoniously accounted for by mean air temperature in the week preceding sampling. Sites with intermediate mean temperatures had more phorid fly individuals and higher richness. Communities were more even at urban sites with lower minimum temperatures and sites located further away from natural areas, suggesting that communities separated from natural source populations may be more homogenized. Species composition was best explained by minimum temperature. Inasmuch as warmer areas within cities can predict future effects of climate change, phorid fly communities are likely to shift nonlinearly under future climates in more natural areas. Exhaustive surveys of biotic communities within cities, such as the one we describe here, can provide baselines for determining the effects of urban and global climate warming as they intensify.

Mystery mushroom malingerers: Megaselia marquezi Hartop et al. 2015 (Diptera: Phoridae).

A mysterious female phorid fly, known for many years to be associated with fungal sporophores ("mushrooms") is identified as Megaselia marquezi Hartop et al. 2015. Male and female flies were collected emerging from the fungus Psathyrella candolleana (Fr.) Maire, and females were observed swarming over the sporophores.

Baby Killers: Documentation and Evolution of Scuttle Fly (Diptera: Phoridae) Parasitism of Ant (Hymenoptera: Formicidae) Brood.

BACKGROUND: Numerous well-documented associations occur among species of scuttle flies (Diptera: Phoridae) and ants (Hymenoptera: Formicidae), but examples of brood parasitism are rare and the mechanisms of parasitism often remain unsubstantiated. NEW INFORMATION: We present two video-documented examples of ant brood (larvae and pupae) parasitism by scuttle flies. In footage from Estação Biológica de Boracéia in Brazil, adult females of Ceratoconus setipennis Borgmeier can be seen attacking workers of Linepithema humile (Mayr) species group while they are carrying brood, and ovipositing directly onto brood in the nest. In another remarkable example, footage from the Soltis Center, near Peñas Blancas in Costa Rica, shows adult females of an unidentified species of the Apocephalus grandipalpus Borgmeier group mounting Pheidole Westwood brood upside-down and ovipositing while the brood are being transported by workers. Analysis of evolutionary relationships (in preparation) among Apocephalus Coquillett species shows that this is a newly derived behavior within the genus, as the A. grandipalpus group arises within a group of adult ant parasitoids. In contrast, relationships of Ceratoconus Borgmeier have not been studied, and the lifestyles of the other species in the genus are largely unknown.

Flies from L.A., The Sequel: A further twelve new species of Megaselia (Diptera: Phoridae) from the BioSCAN Project in Los Angeles (California, USA).

BACKGROUND: Presented are continued results from the BioSCAN Project, an urban biodiversity study sampling primarily from private backyards in Los Angeles, California (USA). Presented are continued results from the BioSCAN Project, an urban biodiversity study sampling primarily from private backyards in Los Angeles, California (USA). NEW INFORMATION: Twelve new species of Megaselia (Diptera: Phoridae) are described: M. baileyae, M. friedrichae, M. gonzalezorum, M. joanneae, M. losangelensis, M. phyllissunae, M. pongsaiae, M. shatesae, M. stoakesi, M. studentorum, M. voluntariorum, M. wongae.

Opportunity in our Ignorance: Urban Biodiversity Study Reveals 30 New Species and One New Nearctic Record for Megaselia (Diptera: Phoridae) in Los Angeles (California, USA).

An urban biodiversity study sampling primarily from private backyards in Los Angeles, California (USA), reveals the presence of fifty-six species of Megaselia within the first few months of sampling. Thirty of these are described as new to science: M. armstrongorum, M. bradyi, M. brejchaorum, M. carthayensis, M. ciancii, M. creasoni, M. defibaughorum, M. donahuei, M. francoae, M. fujiokai, M. hardingorum, M. heini, M. hentschkeae, M. hoffmanorum, M. hoggorum, M. hoguei, M. isaacmajorum, M. kelleri, M. lombardorum, M. marquezi, M. mikejohnsoni, M. oxboroughae, M. pisanoi, M. renwickorum, M. rodriguezorum, M. sacatelensis, M. seaverorum, M. sidneyae, M. steptoeae, and M. wiegmanae. M. largifrontalis is newly reported from the Nearctic Region. The implications these findings have for future taxonomic work in Megaselia, particularly in urban areas, are discussed.

The tip of the iceberg: a distinctive new spotted-wing Megaselia species (Diptera: Phoridae) from a tropical cloud forest survey and a new, streamlined method for Megaselia descriptions.

A new Megaselia species, M.shadeae, with a large, central, pigmented and bubble-like wing spot and a greatly enlarged radial wing vein fork, is described from Zurquí de Moravia, Costa Rica. As part of the Zurquí All Diptera Biodiversity Inventory (ZADBI) project, it represents the first of an incredible number of new phorid species to be described from this one Costa Rican cloud forest site. A new, streamlined method of description for species of this enormous genus of phorid flies is presented.