New Colombian Goja Navs (Psocodea: 'Psocoptera: Epipsocidae) with peculiar genitalia, and the first Goja with brachypterous male, from Oaxaca, Mexico.

Eight species of Goja are here described and illustrated, seven of them are from Colombia, they present peculiar phallosomes and constitute assemblages of three and four related species each, they are: G. andina n. sp., G. garridoi n. sp., G. meremberg n. sp., G. risaraldensis n. sp., G. tenerife n. sp., G. vallecaucana n. sp. and G. vavilovi n. sp. The other species, G. sierrajuarez n. sp., collected in the Sierra Jurez of Oaxaca, Mexico, is the first species of Goja, in which the male is brachypterous. A key to the identification of the males here treated is included, as well as a cladistic analysis of the species of Goja known to date.

Global population genetic structure and demographic trajectories of the black soldier fly, Hermetia illucens.

BACKGROUND: The black soldier fly (Hermetia illucens) is the most promising insect candidate for nutrient-recycling through bioconversion of organic waste into biomass, thereby improving sustainability of protein supplies for animal feed and facilitating transition to a circular economy. Contrary to conventional livestock, genetic resources of farmed insects remain poorly characterised. We present the first comprehensive population genetic characterisation of H. illucens. Based on 15 novel microsatellite markers, we genotyped and analysed 2862 individuals from 150 wild and captive populations originating from 57 countries on seven subcontinents. RESULTS: We identified 16 well-distinguished genetic clusters indicating substantial global population structure. The data revealed genetic hotspots in central South America and successive northwards range expansions within the indigenous ranges of the Americas. Colonisations and naturalisations of largely unique genetic profiles occurred on all non-native continents, either preceded by demographically independent founder events from various single sources or involving admixture scenarios. A decisive primarily admixed Polynesian bridgehead population serially colonised the entire Australasian region and its secondarily admixed descendants successively mediated invasions into Africa and Europe. Conversely, captive populations from several continents traced back to a single North American origin and exhibit considerably reduced genetic diversity, although some farmed strains carry distinct genetic signatures. We highlight genetic footprints characteristic of progressing domestication due to increasing socio-economic importance of H. illucens, and ongoing introgression between domesticated strains globally traded for large-scale farming and wild populations in some regions. CONCLUSIONS: We document the dynamic population genetic history of a cosmopolitan dipteran of South American origin shaped by striking geographic patterns. These reflect both ancient dispersal routes, and stochastic and heterogeneous anthropogenic introductions during the last century leading to pronounced diversification of worldwide structure of H. illucens. Upon the recent advent of its agronomic commercialisation, however, current human-mediated translocations of the black soldier fly largely involve genetically highly uniform domesticated strains, which meanwhile threaten the genetic integrity of differentiated unique local resources through introgression. Our in-depth reconstruction of the contemporary and historical demographic trajectories of H. illucens emphasises benchmarking potential for applied future research on this emerging model of the prospering insect-livestock sector.

New species of Goja Navás (Psocodea: 'Psocoptera': Epipsocidae) from Colombia, Mexico and Peru.

Upon examination of epipsocid specimens recently collected in natural areas of Colombia, Mexico and Peru, 17 new species of Goja were found. These are here described and illustrated. An updated key for the identification of males of the described species of Goja is presented.

Identification keys to the Anopheles mosquitoes of South America (Diptera: Culicidae). IV. Adult females.

BACKGROUND: Morphological identification of adult females of described species of the genus Anopheles Meigen, 1818 in South America is problematic, but necessary due to their differing roles in the transmission of human malaria. The increase in the number of species complexes uncovered by molecular taxonomy challenges accurate identification using morphology. In addition, the majority of newly discovered species have not been formally described and in some cases the identities of the nominotypical species of species complexes have not been resolved. Here, we provide an up-to-date key to identify Neotropical Anopheles species using female external morphology and employing traditionally used and new characters. METHODS: Morphological characters of the females of South American species of the genus Anopheles were examined and employed to construct a species/group identification key. Photographs of key characters were obtained using a digital Canon Eos T3i, attached to a microscope. The program Helicon Focus was used to build single in-focus images by stacking multiple images of the same structure. RESULTS: A morphological identification key to the adult females of species of the genus Anopheles described in South America is presented. Definitions and illustrations of the key characters are provided to facilitate use of key. CONCLUSIONS: Identification of species of the genus Anopheles based on female morphology is challenging because some key characters can be variable and overlapping among species. In addition, the majority of key characters are linked to color and shape of scales, their distribution on the head, scutum, abdomen, maxillary palpi, labium and legs, and pattern of pale and dark scales on dorsal and ventral surfaces of the wing veins. Thus, it is understandable that a specimen needs to be in good condition to be accurately identified. Morphologically similar species, such as those of the Konderi, Oswaldoi, Nuneztovari, Benarrochi and Albitarsis Complexes, and the Triannulatus and Strodei Groups, among others, cannot be accurately identified using characters included in the key. Further investigation will be required to exploit morphological characteristics for identification of members of those complexes, with formal description of new species.

Identification keys to the Anopheles mosquitoes of South America (Diptera: Culicidae). II. Fourth-instar larvae.

BACKGROUND: Accurate species identification of South American anophelines using morphological characters of the fourth-instar larva is problematic, because of the lack of up-to-date identification keys. In addition, taxonomic studies, employing scanning electron microscopy of the eggs and DNA sequence data, have uncovered multiple complexes of morphologically similar species, and resulted in the resurrection of other species from synonymy, mainly in the subgenus Nyssorhynchus. Consequently, the identification keys urgently need to be updated to provide accurate morphological tools to identify fourth-instar larvae of all valid species and species complexes. METHODS: Morphological characters of the fourth-instar larvae of South American species of the genus Anopheles were examined and employed to elaborate a fully illustrated identification key. For species for which no specimens were available, illustrations were based on published literature records. RESULTS: A fully illustrated key to the fourth-instar larvae of South American species of the genus Anopheles (Diptera: Culicidae) is presented. Definitions of the morphological terms used in the key are provided and illustrated. CONCLUSIONS: Morphological identification of South American Anopheles species based on the fourth-instar larvae has been updated. Characters of the spiracular apparatus were determined useful for the identification of morphologically similar species, in the Strodei Group and some taxa in the Myzorhynchella Section. The single versus branched abdominal seta 6-IV used to differentiate Myzorhynchella species from other Nyssorhynchus species was shown to be variable in Myzorhynchella species. Also, the abdominal setae 1-IV,V of Anopheles atacamensis and Anopheles pictipennis were shown to be slightly serrate at the edges. Recognition of this character is important to avoid inaccurate identification of these species as members of the subgenus Anopheles.

Identification keys to the Anopheles mosquitoes of South America (Diptera: Culicidae). I. Introduction.

BACKGROUND: The worldwide genus Anopheles Meigen, 1918 is the only genus containing species evolved as vectors of human and simian malaria. Morbidity and mortality caused by Plasmodium Marchiafava & Celli, 1885 is tremendous, which has made these parasites and their vectors the objects of intense research aimed at mosquito identification, malaria control and elimination. DNA tools make the identification of Anopheles species both easier and more difficult. Easier in that putative species can nearly always be separated based on DNA data; more difficult in that attaching a scientific name to a species is often problematic because morphological characters are often difficult to interpret or even see; and DNA technology might not be available and affordable. Added to this are the many species that are either not yet recognized or are similar to, or identical with, named species. The first step in solving Anopheles identification problem is to attach a morphology-based formal or informal name to a specimen. These names are hypotheses to be tested with further morphological observations and/or DNA evidence. The overarching objective is to be able to communicate about a given species under study. In South America, morphological identification which is the first step in the above process is often difficult because of lack of taxonomic expertise and/or inadequate identification keys, written for local fauna, containing the most consequential species, or obviously, do not include species described subsequent to key publication. METHODS: Holotypes and paratypes and other specimens deposited in the Coleção Entomológica de Referência, Faculdade de Saúde Pública (FSP-USP), Museo de Entomología, Universidad del Valle (MUSENUV) and the US National Mosquito Collection, Smithsonian Institution (USNMC) were examined and employed to illustrate the identification keys for female, male and fourth-instar larvae of Anopheles. RESULTS: We presented, in four concurrent parts, introduction and three keys to aid the identification of South American Anopheles based on the morphology of the larvae, male genitalia and adult females, with the former two keys fully illustrated. CONCLUSIONS: Taxonomic information and identification keys for species of the genus Anopheles are updated. The need for further morphology-based studies and description of new species are reinforced.

Identification key to the Anopheles mosquitoes of South America (Diptera: Culicidae). III. Male genitalia.

BACKGROUND: Accurate identification of the species of Anopheles Meigen, 1818 requires careful examination of all life stages. However, morphological characters, especially those of the females and fourth-instar larvae, show some degree of polymorphism and overlap among members of species complexes, and sometimes even within progenies. Characters of the male genitalia are structural and allow accurate identification of the majority of species, excluding only those in the Albitarsis Complex. In this key, based on the morphology of the male genitalia, traditionally used important characters are exploited together with additional characters that allow robust identification of male Anopheles mosquitoes in South America. METHODS: Morphological characters of the male genitalia of South American species of the genus Anopheles were examined and employed to construct a comprehensive, illustrated identification key. For those species for which specimens were not available, illustrations were based on published illustrations. Photographs of key characters of the genitalia were obtained using a digital Canon Eos T3i attached to a light Diaplan Leitz microscope. The program Helicon Focus was used to build single in-focus images by stacking multiple images of the same structure. RESULTS: An illustrated key to South American species of Anopheles based on the morphology of the male genitalia is presented, together with a glossary of morphological terms. The male genitalia of type-specimens of previously poorly documented species were also examined and included in the key, e.g. Anopheles (Anopheles) tibiamaculatus (Neiva, 1906) which has a unique quadrangular-shaped aedeagus with an apical opening. CONCLUSIONS: Male genitalia of South American species of Anopheles possess robust characters that can be exploited for accurate species identification. Distortion that can occur during the dissection and mounting process can obstruct accurate identification; this is most evident with inadvertent damage or destruction of unique features and interferes with correctly assigning shapes of the features of the ventral claspette. In some species, the shape, and anatomical details of the aedeagus also need to be examined for species identification. For members of the Myzorhynchella Series, both ventral and dorsal claspettes possess multiple characteristics that are herein used as reliable characters for species identification.

Saving DNA from museum specimens: The success of DNA mini-barcodes in haplotype reconstruction in the genus Anastrepha (Diptera: Tephritidae).

The fragmentation of DNA in historical specimens is very common, so obtaining sequences that allow molecular identification and the study of diversity is quite challenging. In this study, we used preserved and fresh specimens of the fruit fly genus Anastrepha, a genus of economic impact of fruit crops of the Neotropic. From these specimens, we evaluated: (1) the success PCR amplification rates of mini-barcodes fragments of the cytochrome c oxidase subunit I (COI) gene, and (2) the usefulness of mini-barcodes in the reconstruction of haplotypes for the identification of species and the diversity analysis. We used 93 specimens from 12 species, which had been preserved in 70% ethanol for more than 20 years. Internal primers were designed in the COI region and primers available in the literature were also evaluated. We obtained amplifications for 62.36% of the samples processed, and reconstructed haplotypes between 171 bp and 632 bp. Variable amplification rates between combinations of primers and between species were obtained, and molecular identification of some museum specimens was achieved. It was also possible to compare the haplotypes obtained in four species from which both fresh and museum samples were available. Our results also show the importance of the adjustment of the primers for the amplification, allowing to amplify fragments of up to 400 bp. The use available resources in biological collections is key to increasing knowledge of species of interest, and by means of the amplification of mini-barcodes, short sequences can be obtained that allow the molecular identification of specimens and the reconstruction of haplotypes with multiple purposes.

New species of Euplocania Enderlein (Psocodea: 'Psocoptera': Psocomorpha), in three species groups, from Colombia and Ecuador.

Eight species of Euplocania from Colombia, in species groups Bonaverensis, Enderleini and Patinoi, are here described and illustrated. They raise to 40 the number of species described in the genus, 27 of which are found in Colombia. Of the Colombian species, one is shared with Ecuador and one is shared with Brazil and Peru. A key to identify the males of the species here described is included.

New species and records of Cladiopsocus Roesler (Psocodea: 'Psocoptera': Psocomorpha: Cladiopsocidae) from Colombia.

Thirteen species of Cladiopsocus Roesler, from the Colombian departments of Caquetá, Chocó, Magdalena, Nariño, Norte de Santander and Putumayo are here described and illustrated. New records of eight described species of Cladiopsocus, from the departments of Caquetá, Cauca, Quindío and Valle del Cauca are included. The number of species in the genus is increased to 41, of which 27 are Colombian. An updated identification key to the males of the genus is included, as well as a map of species distribution.

Five new species of the genus Euplocania Enderlein (Psocodea, 'Psocoptera', Psocomorpha, Ptiloneuridae) from Colombia.

Five new species of Euplocania from Colombia belonging in four species groups are here described and illustrated. They increase to 22 the number of species described in the genus, thirteen of which are endemic to Colombia, with one species shared with Brazil and Peru. Three new species groups of Euplocania are here diagnosed. A key to the identification of males of Colombian Euplocania is included.

Genetic diversity and population structure of Anastrepha striata (Diptera: Tephritidae) in three natural regions of southwestern Colombia using mitochondrial sequences.

Anastrepha striata is widely distributed across the Americas and is a pest of economically important crops, especially crops of the Myrtaceae family. Insect population structures can be influenced by the presence of physical barriers or characteristics associated with habitat differences. This study evaluated the effect of the Western Andes on the population structure of A. striata. Individuals were collected from Psidium guajava fruits from three natural regions of southwestern Colombia (Pacific Coast, mountainous region and the inter-Andean valley of the Cauca River). Based on a 1318 bp concatenated of the genes Cytochrome Oxidase subunit I (COI) and NADH dehydrogenase subunit 6 (ND6), 14 haplotypes with few changes among them (between 1 and 3) were found. There was only one dominant haplotype in all three regions. No genetic structure associated with the three eco-geographical regions of the study was found. Moreover, the Western Andes are not an effective barrier for the genetic isolation of the populations from the Pacific Coast compared with the inter-Andean valley populations. This genetic homogeneity could be partially due to anthropogenic intervention, which acts as a dispersal agent of infested fruits. Another hypothesis to explain the lack of structure would be the relatively recent arrival of A. striata to the region, as indicated by an analysis of the demographic history, which reveals a process of population expansion. This study represents the first attempt to understand the population genetics of A. striata in Colombia and could contribute to the integral management of this pest.

New species of Cladiopsocus Roesler (Psocodea: 'Psocoptera': Psocomorpha: Cladiopsocidae) from the Colombian Amazonia.

Ten species of Cladiopsocus Roesler were identified among recent collections from the Colombian departments of Amazonas and Putumayo. Eight new species were found, five from Putumayo and three from Amazonas, that are here described and illustrated. C. ramulosus (Enderlein) and C. domesticus (New) were found in the Putumayo department. The number of species in the genus is increased to 28, with two from Angola, and 26 from the Neotropics. An identification key to the males of the neotropical species is included.

New genera of Lachesillidae (Psocodea: 'Psocoptera': Eolachesillinae: Graphocaeciliini) from Valle del Cauca, Colombia.

Two new related Colombian genera of Lachesillidae, including three new species, from the Department of Valle del Cauca, are here described and illustrated. They belong in the tribe Graphocaeciliini (Eolachesillinae). The new genera differ from the other genera in the tribe lacking ocelli, by characters of the male clunium, phallosome, endophallus, paraprocts, and female subgenital plate, gonapophyses and IX sternum. The types are deposited in the Entomological Museum of the Universidad del Valle (MUSENUV), in Santiago de Cali, Colombia.

A new species of Lixophaga Townsend (Diptera: Tachinidae) from Colombia, a parasitoid of Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae).

A new species of Lixophaga Townsend (Diptera: Tachinidae) from Colombia, Lixophaga puscolulo Carrejo & Woodley, sp. nov., is described and illustrated. It is a parasitoid of the tomato fruit borer, Neoleucinodes elegantalis (Guenée) (Lepidoptera: Crambidae), an insect pest of Solanum quitoense Lam., in Colombia. Aspects of its biology are briefly discussed.

Confirmation of Anopheles (Anopheles) calderoni Wilkerson, 1991 (Diptera: Culicidae) in Colombia and Ecuador through molecular and morphological correlation with topotypic material

The morphologically similar taxa Anopheles calderoni, Anopheles punctimacula, Anopheles malefactor and Anopheles guarao are commonly misidentified. Isofamilies collected in Valle de Cauca, Colombia, showed morphological characters most similar to An. calderoni, a species which has never previously been reported in Colombia. Although discontinuity of the postsubcostal pale spots on the costa (C) and first radial (R1) wing veins is purportedly diagnostic for An. calderoni, the degree of overlap of the distal postsubcostal spot on C and R1 were variable in Colombian specimens (0.003-0.024). In addition, in 98.2 percent of larvae, seta 1-X was located off the saddle and seta 3-C had 4-7 branches in 86.7 percent of specimens examined. Correlation of DNA sequences of the second internal transcribed spacer and mtDNA cytochrome c oxidase subunit I gene (COI) barcodes (658 bp of the COI gene) generated from Colombian progeny material and wild-caught mosquitoes from Ecuador with those from the Peruvian type series of An. calderoni confirmed new country records. DNA barcodes generated for the closely related taxa, An. malefactor and An. punctimacula are also presented for the first time. Examination of museum specimens at the University of the Valle, Colombia, revealed the presence of An. calderoni in inland localities across Colombia and at elevations up to 1113 m.

Confirmation of Anopheles (Anopheles) calderoni Wilkerson, 1991 (Diptera: Culicidae) in Colombia and Ecuador through molecular and morphological correlation with topotypic material.

The morphologically similar taxa Anopheles calderoni, Anopheles punctimacula, Anopheles malefactor and Anopheles guarao are commonly misidentified. Isofamilies collected in Valle de Cauca, Colombia, showed morphological characters most similar to An. calderoni, a species which has never previously been reported in Colombia. Although discontinuity of the postsubcostal pale spots on the costa (C) and first radial (R1) wing veins is purportedly diagnostic for An. calderoni, the degree of overlap of the distal postsubcostal spot on C and R1 were variable in Colombian specimens (0.003-0.024). In addition, in 98.2% of larvae, seta 1-X was located off the saddle and seta 3-C had 4-7 branches in 86.7% of specimens examined. Correlation of DNA sequences of the second internal transcribed spacer and mtDNA cytochrome c oxidase subunit I gene (COI) barcodes (658 bp of the COI gene) generated from Colombian progeny material and wild-caught mosquitoes from Ecuador with those from the Peruvian type series of An. calderoni confirmed new country records. DNA barcodes generated for the closely related taxa, An. malefactor and An. punctimacula are also presented for the first time. Examination of museum specimens at the University of the Valle, Colombia, revealed the presence of An. calderoni in inland localities across Colombia and at elevations up to 1113 m.