Viruses encoding a replication-associated protein (Rep) within a covalently closed, single-stranded (ss)DNA genome are among the smallest viruses known to infect eukaryotic organisms, including economically valuable agricultural crops and livestock. Although circular Rep-encoding ssDNA (CRESS DNA) viruses are a widespread group for which our knowledge is rapidly expanding, biased sampling toward vertebrates and land plants has limited our understanding of their diversity and evolution. Here, we screened terrestrial arthropods for CRESS DNA viruses and report the identification of 44 viral genomes and replicons associated with specimens representing all three major terrestrial arthropod lineages, namely Euchelicerata (spiders), Hexapoda (insects), and Myriapoda (millipedes). We identified virus genomes belonging to three established CRESS DNA viral families (Circoviridae, Genomoviridae, and Smacoviridae); however, over half of the arthropod-associated viral genomes are only distantly related to currently classified CRESS DNA viral sequences. Although members of viral and satellite families known to infect plants (Geminiviridae, Nanoviridae, Alphasatellitidae) were not identified in this study, these plant-infecting CRESS DNA viruses and replicons are transmitted by hemipterans. Therefore, members from six out of the seven established CRESS DNA viral families circulate among arthropods. Furthermore, a phylogenetic analysis of Reps, including endogenous viral sequences, reported to date from a wide array of organisms revealed that most of the known CRESS DNA viral diversity circulates among invertebrates. Our results highlight the vast and unexplored diversity of CRESS DNA viruses among invertebrates and parallel findings from RNA viral discovery efforts in undersampled taxa.

Correct identification of forensically important insects, such as flies in the family Calliphoridae, is a crucial step for them to be used as evidence in legal investigations. Traditional identification based on morphology has been effective, but has some limitations when it comes to identifying immature stages of certain species. DNA-barcoding, using COI, has demonstrated potential for rapid and accurate identification of Calliphoridae, however, this gene does not reliably distinguish among some recently diverged species, raising questions about its use for delimitation of species of forensic importance. To facilitate DNA based identification of Calliphoridae in the Caribbean we developed a vouchered reference collection from across the region, and a DNA sequence database, and further added the nuclear ITS2 as a second marker to increase accuracy of identification through barcoding. We morphologically identified freshly collected specimens, did phylogenetic analyses and employed several species delimitation methods for a total of 468 individuals representing 19 described species. Our results show that combination of COI + ITS2 genes yields more accurate identification and diagnoses, and better agreement with morphological data, than the mitochondrial barcodes alone. All of our results from independent and concatenated trees and most of the species delimitation methods yield considerably higher diversity estimates than the distance based approach and morphology. Molecular data support at least 24 distinct clades within Calliphoridae in this study, recovering substantial geographic variation for Lucilia eximia, Lucilia retroversa, Lucilia rica and Chloroprocta idioidea, probably indicating several cryptic species. In sum, our study demonstrates the importance of employing a second nuclear marker for barcoding analyses and species delimitation of calliphorids, and the power of molecular data in combination with a complete reference database to enable identification of taxonomically and geographically diverse insects of forensic importance.

The Neotropical family Mesembrinellidae is revised. A total of 53 valid, extant species are included in the family, including 15 described as new and 38 redescribed based on study of type and non-type material and of the literature. A total of 18 primary types were examined. An additional ca. 2300 specimens, belonging to 47 species, were studied in detail, including dissection and photographic documentation of terminalia, with many females illustrated for the first time. Keys to subfamilies, genera, species-groups and species are provided. Type specimens of six species housed in South American institutions could not be obtained for study, i.e., M. bequaerti Séguy, 1925 and the five recently described species M. andina (Wolff et al., 2014), M. carvalhoi (Wolff et al., 2013b), M. cordillera (Wolff & Ramos-Pastrana in Wolff et al., 2017), M. obscura (Wolff in Wolff et al., 2017) and Laneella patriciae (Wolff, 2013). We accept the synonymy, proposed by previous authors, of Eumesembrinella Townsend, 1931 with Mesembrinella Giglio-Tos, 1893. In addition, we synonymize the genera Albuquerquea Mello, 1967, Giovanella Bonatto in Bonatto & Marinoni, 2005, Henriquella Bonatto in Bonatto & Marinoni, 2005, Huascaromusca Townsend, 1918 and Thompsoniella Guimarães, 1977 with Mesembrinella Giglio-Tos, 1893, synn. nov., retaining three valid genera in the family: Laneella Mello, 1967, Mesembrinella and Souzalopesiella Guimarães, 1977. Laneella nigripes Guimarães, 1977 and Mesembrinella bellardiana Aldrich, 1922 are fixed as the type species of the genera Laneella Mello, 1967 and Mesembrinella Giglio-Tos, 1893, respectively, under Article 70.3 of the ICZN Code. We separate Mesembrinella into the following species-groups: M. latifrons (Mello, 1967), M. spicata Aldrich, 1925, M. bolivar (Bonatto in Bonatto & Marinoni, 2005), M. aeneiventris (Wiedemann, 1830), M. bicolor (Fabricius, 1805), and M. anomala (Guimarães, 1977). The following 15 new species are described: Laneella fusconitida Whitworth, sp. nov. from Costa Rica, Ecuador and Venezuela, Laneella fuscosquamata Whitworth, sp. nov. from Guatemala and Mexico, Laneella purpurea Whitworth, sp. nov. from Costa Rica, Mesembrinella bullata Whitworth, sp. nov. from Bolivia, Mesembrinella chantryi Whitworth, sp. nov. from French Guiana and Brazil, Mesembrinella epandrioaurantia Whitworth, sp. nov. from Venezuela, Mesembrinella guaramacalensis Whitworth, sp. nov. from Venezuela, Mesembrinella longicercus Whitworth, sp. nov. from Bolivia, Mesembrinella mexicana Whitworth, sp. nov. from Mexico, Mesembrinella nigrocoerulea Whitworth, sp. nov. from Costa Rica, Ecuador and Venezuela, Mesembrinella serrata Whitworth, sp. nov. from Peru, Mesembrinella velasquezae Whitworth, sp. nov. from Venezuela, Mesembrinella violacea Whitworth, sp. nov. from Costa Rica, Mesembrinella woodorum Whitworth, sp. nov. from Ecuador, and Mesembrinella zurquiensis Whitworth, sp. nov. from Costa Rica. Mesembrinella abaca Hall, 1948 is proposed as a junior synonym of Mesembrinella socors (Walker, 1861), syn. nov. Lectotypes are designated for Dexia randa Walker, 1849 (now Mesembrinella) and Mesembrinella pictipennis Aldrich, 1922. We analyze the most extensive DNA-barcode dataset for Mesembrinellidae to date, encompassing the three genera considered valid and including 188 sequences (178 new) from 35 species, with data for 23 species provided for the first time. The topology of the resulting Neighbor-Joining tree is mostly congruent with morphology; however, some species show considerable genetic variation that is not reflected by morphology. Finally, we include a corrigendum to the recent Zootaxa paper on Nearctic Calliphora Robineau-Desvoidy (Diptera: Calliphoridae) by Tantawi et al.

Cochliomyia Townsend includes several abundant and one of the most broadly distributed, blow flies in the Americas, and is of significant economic and forensic importance. For decades, Cochliomyia hominivorax (Coquerel) and Cochliomyia macellaria (Fabricius) have received attention as livestock parasites and primary indicator species in forensic entomology. However, Cochliomyia minima Shannon and Cochliomyia aldrichi Del Ponte have only been subject to basic taxonomy and faunistic studies. Here we present the first complete phylogeny of Cochliomyia including numerous specimens per species, collected from 13 localities in the Caribbean. Four genes, the mitochondrial COI and the nuclear EF-1α, 28S rRNA, and ITS2, were analyzed. While we found some differences among gene trees, a concatenated gene matrix recovered a robustly supported monophyletic Cochliomyia with Compsomyiops Townsend as its sister group and recovered the monophyly of Cochliomyia hominivorax, Cochliomyia macellaria and Cochliomyia minima. Our results support a close relationship between Cochliomyia minima and Cochliomyia aldrichi. However, we found Cochliomyia aldrichi containing Cochliomyia minima, indicating recent speciation, or issues with the taxonomy of the group. We provide basic information on habitat preference, distribution and feeding habits of Cochliomyia minima and Cochliomyia aldrichi that will be useful for future forensic studies in the Caribbean.

The blow fly Cochliomyia minima Shannon is endemic to the Caribbean, and it has great potential for forensic applications because of its abundance and broad distribution in the region. However, its larval stages are unknown. Here, I update previously published identification keys by describing for the first time the morphology of C. minima larvae. The larvae of C. minima are found to be very similar to those of Cochliomyia macellaria F., but the former can be easily identified by the oral sclerite completely pigmented, visible as a spike between mouth hooks. The description of C. minima larvae in this study will be useful to forensic scientists in the Caribbean region.