Redescription of Raillietiella mottae (Raillietiellidae: Pentastomida) a generalist parasite of insectivorous lizards: phenotypic plasticity revealed by molecular data.

Pentastomida are lung parasites of a variety of hosts, mainly reptiles. Raillietiella is the genus with the largest number of species, whose taxonomic identification is still largely based on morphological data alone. In this classification, differences divide the genus into morphotypic groups. The Raillietiella parasites of insectivorous lizards, for example, are divided into "Group I" with species with hooks with sharp tips and "Group II" with posterior hooks with blunt tips. In the present work, we used molecular analysis to infer the morphological variation in hooks and size of specimens of Raillietiella mottae, parasites of several species of insectivorous lizards. Our results demonstrate that morphological differences comprise a phenotypic plasticity of a single species, whose variation is associated with host species, with females being larger and with hooks with blunt tips in nocturnal geckos and smaller ones with sharp tips in diurnal insectivorous lizards of different species, sizes, and habitats.

First report of root-knot nematode, Meloidogyne javaniva, infecting Stachys byzantina on São Paulo, Brazil.

Stachys byzantina belongs to the Labiatae and is known by the names "peixinho-da-horta" (Brazil) and "lamb's ear" (USA). Its importance is associated with its medicinal properties (Bahadori et al. 2020) and nutritional aspects (Milião et al. 2022). Root-knot nematodes cause severe damage to plants and suppress production. In January 2021, plants of S. byzantina in the municipality of Jaboticabal (21°14'38.7"S, 48°17'10.6"W) showed symptoms of reduced growth, yellowed leaves and the presence of galls in the roots. Initially, samples of roots from a S. byzantina were analyzed at the Nematology Laboratory (LabNema/UNESP), Jaboticabal, Brazil, estimating 20,000 eggs and juveniles of Meloidogyne sp. in 10 g of roots. To confirm the host ability of the species, a pathogenicity test was performed using Koch's postulate. For this purpose, the test was conducted in a greenhouse where 3,000 eggs and second-stage juveniles (J2) were inoculated onto three plants (n=3) of S. byzantina. After 90 days, the inoculated plants showed the same symptoms as those observed in the field. No symptom or nematode was detected in the uninoculated plant (control). Nematodes were extracted from the roots of inoculated plants and quantified. The perineal pattern of females (n=10) (Netscher and Taylor, 1974) and the labial region of males (n=10) (Eisenback and Hirschmann, 1981) were analyzed and compared with the morphological characteristics of the original description of the species (Chitwood, 1949). For analysis based on esterase isozyme phenotype, the α-method of Esbenshade and Triantaphyllou (1990) was used, and females (n=7) were examined. To confirm identification, whole genomic DNA from an adult female (n=1) was extracted using the Qiagen DNeasy® Blood & Tissue Kit and this sample was used for both genetic sequencing and the sequence-characterized amplified region techniques (SCAR). PCR amplifications were performed for the 18s rRNA gene using primers 988F and 1912R from Holterman et al (2006). Our sequence was deposited in GenBank (NCBI) under the identifier OP422209. Finally, species-specific SCAR primers (Fjav/Rjav, Me-F/Me-R, and Finc-F/Finc-R) designed by Zijlstra (2000) were used to identify Meloidogyne spp. Koch's postulate analysis yielded the following results: (n=1) 9,280 eggs and J2 (Reproduction factor, RF = 33.09); (n=2) 111,720 eggs and J2 (RF = 37.24); (n=3) 59,700 eggs and J2 (RF = 19.9) (RF mean = 30.08). The following characteristics were observed in the perineal region of females: Low and rounded trapezoidal dorsal arch with two distinct lateral lines clearly separating the dorsal and ventral arch regions, similar to the morphological features of the species description by Chitwood (1949). Males had a convex labial plate with a non-raised labial disk joining the submedial labia, a non-rugged labial region, the basal tubercles were usually wider than high, and a rounded tail tip (Eisenback and Hirschmann 1981). The α-esterase enzyme profile showed the J3 phenotype typical of M. javanica (Rm [×100] = 46.0, 54.5, and 58.9). The 18s rRNA sequences grouped Meloidogyne sp. with species such as M. enterolobii, M. incognita, and M. javanica. A DNA fragment of about 700 bp was amplified with Mj (Fjav/Rjav) primers, but not with Me (Me-F/Me-R) and Mi (Finc-F/Finc-R) primers, which confirmed the identification of M. javanica. Accurate identification and characterization of the occurrence of new hosts of M. javanica will allow us to determine the range and geographic distribution of the species. This is the first report on the occurrence of M. javanica on S. byzantina in Brazil. This report is important so that management strategies can be applied to prevent the spread of the pest to other areas.

Evaluation of partial 12S rRNA, 16S rRNA, COI and Cytb gene sequence datasets for potential single DNA barcode for hylids (Anura: Hylidae).

We evaluated the extent of intraspecific and interspecific genetic distances and the effectiveness of predefined threshold values using the main genes for estimates of biodiversity and specimens' identification in anurans. Partial sequences of the mitochondrial genes for small (12S) and large (16S) ribosomal subunits, cytochrome c oxidase subunit I (COI) and Cytochrome b (Cytb) of the family Hylidae were downloaded from GenBank and curated for length, coverage, and potential contaminations. We performed analyses for all sequences of each gene and the same species present in these datasets by distance and tree (monophyly)-based evaluations. We also evaluated the ability to identify specimens using these datasets applying "nearest neighbor" (NN), "best close match" (BCM) and "BOLD ID" tests. Genetic distance thresholds were generated by the function 'threshVal' and "localMinima" from SPIDER package and traditional threshold values (1%, 3%, 6% and 10%) were also evaluated. Coding genes, especially COI, had a better identification capacity than non-coding genes on barcoding gap and monophyly analysis and NN, BCM, BOLD ID tests. Considering the multiple factors involved in global DNA barcoding evaluations, we present a critical assessment of the use of these genes for biodiversity estimation and specimens' identification in anurans (e.g. hylids).

Argia koroivarum sp. nov. (Odonata: Coenagrionidae) from Minas Gerais state, Southeastern Brazil.

Argia koroivarum sp. nov. (BRAZIL, Minas Gerais state, São Roque de Minas, Parque Nacional Serra da Canastra, 9.iv.2019, (-20.2323, -46.6084, 1306m asl), D.S. Vilela, R. Guillermo, R. Koroiva leg., Laboratory of Ecological Studies on Ethology and Evolution (LESTES), Uberaba, Minas Gerais State, Brazil) is described, illustrated, and diagnosed based on specimens collected in Minas Gerais state, Brazil. The new species can be diagnosed from its congeners by the morphology of male ligula, dorsal branch of paraproct, and of female mesostigmal lobes.

A new tiny toad species of Amazophrynella (Anura: Bufonidae) from east of the Guiana Shield in Amazonia, Brazil.

The combination of different approaches has successfully delimited new species within many Neotropical species complexes traditionally classified as a single nominal organism. Recent studies have shown that the Amazonian endemic genus Amazophrynella, currently composed of 12 small-sized species, could harbor several additional species. Based on morphology and molecular data, we describe a new species of Amazophrynella from east of the Guiana Shield, in Pará state, Brazil. The new species is characterized by having one of the biggest size of the genus (SVL of males 16.0-17.8 mm and females 22.9-24.4 mm), presence of a large palmar tubercle (occupying 2/4 of the palmar surface), 5.6-8.1% uncorrected p-distance from its sister clade (including A. teko, A. sp.1, and A. manaos) for the 16S mitochondrial gene, and 8.8% for the COI. The new species described here represents a newly discovered lineage. Of the 12 Amazophrynella species currently recognized, two were describe in the last century (A. bokermanni and A. minuta) and the remaining species were recently discovered and described (in the last six years), which underscores the degree to which species richness of Amazophrynella is underestimated.

DNA barcoding for identification of anuran species in the central region of South America.

The use of COI barcodes for specimen identification and species discovery has been a useful molecular approach for the study of Anura. Here, we establish a comprehensive amphibian barcode reference database in a central area of South America, in particular for specimens collected in Mato Grosso do Sul state (Brazil), and to evaluate the applicability of the COI gene for species-level identification. Both distance- and tree-based methods were applied for assessing species boundaries and the accuracy of specimen identification was evaluated. A total of 204 mitochondrial COI barcode sequences were evaluated from 22 genera and 59 species (19 newly barcoded species). Our results indicate that morphological and molecular identifications converge for most species, however, some species may present cryptic species due to high intraspecific variation, and there is a high efficiency of specimen identification. Thus, we show that COI sequencing can be used to identify anuran species present in this region.

The female of Epipleoneura spatulata Rácenis, 1960 (Odonata: Protoneurinae).

Here we describe the hitherto unknown female of Epipleoneura spatulata Rácenis, 1960 and present a table of characters (all illustrated) to distinguish it from other known congeneric females. We suggest that the main diagnostic character of the female of E. spatulata is the shape of posterior margin of prothorax, which is flat medially with smoothly curved erect lateral projections.

Revision of the genus Minagrion Santos, 1965 (Odonata: Coenagrionidae).

Endemic to Brazil, the genus Minagrion Santos, 1965 is revised for the first time. The genus contains five species, and here we establish diagnostic characters, keys and drawings for both sexes. Furthermore, we propose the synonymy of M. franciscoi Machado Bedê, 2015 new syn. with M. caldense Santos, 1965.

Heteragrion denisye sp. nov. (Odonata: Zygoptera: Heteragrionidae), a notable species from Serra da Canastra, Minas Gerais, Brazil.

Heteragrion denisye sp. nov. (Zygoptera: Heteragrionidae) is described and diagnosed on specimens collected near a Vereda (i.e. palm swamp) area of the National Park of Serra da Canastra, Minas Gerais, Brazil (-20.2323, -46.6085, 1305 m, 25 x 2018, Vilela, Koroiva, Nobrega Lera leg.). This species is unique within the genus and it is easily distinguished from congeners due to its blue coloration pattern and cerci morphology, which is robust and presents a reduced apical portion, longer on most Heteragrion species.

A further study on Franciscobasis Machado & Bedê, 2016 (Odonata: Coenagrionidae), a newly described genus from Minas Gerais, Brazil.

The genus Franciscobasis Machado & Bedê, 2016 is endemic to the Serra da Canastra National Park in Minas Gerais state, Brazil. Two species of Franciscobasis were described simultaneously with the genus description: F. franciscoi and F. sonia, the latter described only from females. Through morphological and molecular analysis, we investigated if F. sonia may represent the young female of F. franciscoi. Resulting data did not present adequate differences between females to characterize them as different species. Therefore, we suggest that F. sonia is a junior synonym of F. franciscoi, and the female of F. franciscoi goes through a complex ontogenetic color change.

Estimating the barcoding gap in a global dataset of cox1 sequences for Odonata: close, but no cigar.

We evaluated the extent of intraspecific and interspecific genetic distances for two highly diverse infraorders of Odonata: Anisoptera and Zygoptera. All cytochrome c oxidase subunit I sequences (cox1), the region chosen for zoological DNA barcoding, present in GenBank for each infraorder were downloaded and curated. For Anisoptera, the final dataset consisted of 2,961 individual cox1 sequences for 536 species and the equivalent numbers for Zygoptera were 2,477 sequences for 497 species. More than 7 million individual genetic comparisons were made and the results indicated that there is a tendency towards a barcoding gap, but that the size of the gap may not be sufficient to robustly infer identities for some taxa. DNA barcoding may be of less use for some odonate taxa, perhaps pertaining to misidentifications in global databases. However, at local scales or with more confined taxonomical sampling, this tool may yet be beneficial in identifying these charismatic organisms.

DNA Barcodes for Forensically Important Fly Species in Brazil.

Here, we analyze 248 DNA barcode sequences of 35 fly species of forensic importance in Brazil. DNA barcoding can be effectively used for specimen identification of these species, allowing the unambiguous identification of 31 species, an overall success rate of 88%. Our results show a high rate of success for molecular identification using DNA barcoding sequences and open new perspectives for immature species identification, a subject on which limited forensic investigations exist in Tropical regions. We also address the implications of building a robust forensic DNA barcode database. A geographic bias is recognized for the COI dataset available for forensically important fly species in Brazil, with concentration of sequences from specimens collected mainly in sites located in the Cerrado, Mata Atlântica, and Pampa biomes.

Amazonian forest termites: a species checklist from the State of Acre, Brazil / Térmitas da Floresta Amazônica: uma lista de espécies do Estado do Acre, Brasil

Abstract The Brazilian state of Acre is located in the southwestern Amazon and it is characterized by a humid tropical forest vegetation that covers plains and mountains. Up to this point, the composition of termite species in the state is not known. The aim of this study was to provide a checklist of termite species or recognizable taxonomic units for the state of Acre. Sampling was conducted through field expeditions at the Serra do Divisor National Park, Chandless State Park, Humaitá Forest Reserve, and Chico Mendes Environmental Park using a standardized rapid termite inventory protocol in the first two areas and active searching collections in the others, without a specific protocol. This study also included occurrence records published in the scientific literature. A total of 128 species and morphospecies of termites were found in Acre, distributed across 59 genera and four families. The most frequently occurring species in Acre was Heterotermes tenuis (Hagen, 1858). The study also identified six new species records for Brazil. The predominant feeding groups were soil-feeders and wood-feeders, as expected from data obtained from surveys in humid tropical forests. Despite the significant number of new records for Acre (112), it is concluded that a larger sampling effort is still required, as many areas of the state have not yet been studied for termites.

Resumen O estado brasileiro do Acre está localizado no sudoeste da Amazônia e é caracterizado por uma vegetação de floresta tropical úmida que cobre planícies e montanhas. Até então, a composição de espécies de térmitas no estado não é conhecida. O objetivo desse estudo foi construir um checklist de espécies ou unidades taxonômicas reconhecíveis de térmitas para o estado do Acre. A amostragem foi conduzida através de expedições de campo no Parque Nacional da Serra do Divisor, no Parque Estadual Chandless, na Reserva Florestal Humaitá, e no Parque Ambiental Chico Mendes utilizando o protocolo rápido de diversidade de térmitas nas duas primeiras áreas e coletas avulsas nas demais, sem um protocolo específico. Este estudo também incluiu registros de ocorrência publicados na literature científica. Um total de 128 espécies e morfoespécies de térmitas foram encontradas no Acre, distribuídas em 59 gêneros e quatro famílias. A espécie de ocorrência mais frequente no Acre foi Heterotermes tenuis (Hagen, 1858). O estudo também identificou seis novos registros de espécies para o Brasil. Os grupos alimentares predominantes foram os humívoros e xilófagos, como esperado a partir de dados obtidos de pesquisas em florestas tropicais úmidas. Apesar do número significativo de novos registros para o Acre (112), conclui-se que ainda é necessário um esforço amostral maior, uma vez que muitas áreas do estado ainda não foram estudadas para térmitas.

DNA barcoding of odonates from the Upper Plata basin: Database creation and genetic diversity estimation.

We present a DNA barcoding study of Neotropical odonates from the Upper Plata basin, Brazil. A total of 38 species were collected in a transition region of "Cerrado" and Atlantic Forest, both regarded as biological hotspots, and 130 cytochrome c oxidase subunit I (COI) barcodes were generated for the collected specimens. The distinct gap between intraspecific (0-2%) and interspecific variation (15% and above) in COI, and resulting separation of Barcode Index Numbers (BIN), allowed for successful identification of specimens in 94% of cases. The 6% fail rate was due to a shared BIN between two separate nominal species. DNA barcoding, based on COI, thus seems to be a reliable and efficient tool for identifying Neotropical odonate specimens down to the species level. These results underscore the utility of DNA barcoding to aid specimen identification in diverse biological hotspots, areas that require urgent action regarding taxonomic surveys and biodiversity conservation.

Checklist and contribution to the knowledge of the odonatofauna of Paraíba state, Brazil / Checklist e contribuição para o conhecimento da odonatofauna do Estado da Paraíba, Brasil

Abstract: We present the first listing of odonatan species (Insecta: Odonata) that occur in the state of Paraíba, Brazil. There are 49 species and 29 genera registered, making Paraíba the third in number of species among the Brazilian states of northeastern region. The families with the largest number of species were Libellulidae, with 31 species and 15 genera, followed by Coenagrionidae with 11 species and 7 genera. Interior regions of the state are under-sampled, which should still lead to an underestimated number of species. In addition, we present taxonomic notes of two species collected during our expeditions: males of Progomphus dorsopallidus Byers, 1934 and females of Macrothemis griseofrons Calvert, 1909. Here, we detail important characteristics and present figures to aid their morphological identifications.

Resumo: Apresentamos a primeira lista de espécies de libélulas (Insecta: Odonata) que ocorrem no estado da Paraíba, Brasil. Há 49 espécies e 29 gêneros registrados, tornando a Paraíba a terceira em número de espécies entre os estados brasileiros da região Nordeste. As famílias com maior número de espécies foram Libellulidae, com 31 espécies e 15 gêneros, seguido por Coenagrionidae com 11 espécies e 7 gêneros. As regiões do interior do estado estão sub amostradas, o que deve levar a um número subestimado de espécies. Além disso, apresentamos notas taxonômicas de duas espécies coletadas durante nossas expedições: machos de Progomphus dorsopallidus Byers, 1934 e fêmeas de Macrothemis griseofrons Calvert, 1909. Aqui, detalhamos características importantes e apresentamos figuras para auxiliar em suas identificações morfológicas.

Checklist of dragonflies and damselflies (Insecta: Odonata) of the Amazonas state, Brazil / Checklist das libélulas (Insecta: Odonata) do estado do Amazonas, Brasil

Abstract: Here we provide a checklist of the odonates from Amazonas state, Brazil. We registered 324 species and 101 genera, making Amazonas the Brazilian state with the most Odonata species recorded. The families with the highest number of species were Coenagrionidae with 32 genera and 101 species, followed by Libellulidae with 28 genera and 100 species and Gomphidae with 12 genera and 45 species. Some regions of Amazonas state remain poorly explored, such as the southern area, and large municipalities, such as São Gabriel da Cachoeira. This work underlines the importance of the biological diversity from Amazonas state and the Amazonian Biome for Odonata species richness in Brazil and shows that many areas in the world's largest tropical forest have not yet been sampled.

Resumo: Aqui nós apresentamos a lista de espécies de libélulas presentes no estado do Amazonas, Brasil. Nós registramos 324 espécies e 101 gêneros, tornando o estado com o maior número de espécies de libélulas registradas no país. As famílias com maior número de espécies foram Coenagrionidae, com 32 gêneros e 101 espécies, seguido por Libellulidae, com 28 gêneros e 100 espécies e Gomphidae com 12 gêneros e 45 espécies. Regiões do estado do Amazonas permanecem pouco exploradas, como a parte sul e os municípios com grande extensão territorial, como São Gabriel da Cachoeira. Este trabalho reconhece a importância da diversidade biológica do estado do Amazonas e do bioma Amazônico para a riqueza de espécies de Odonata no Brasil e mostra que ainda existem muitas áreas não amostradas na maior floresta tropical do mundo.

Dragonflies and damselflies from the West of Minas Gerais, Brazil: checklist and new records / Libélulas do Oeste de Minas Gerais, Brasil: lista de espécies e novos registros

Abstract: The knowledge about the richness and distribution of Brazilian dragonflies is still being unveiled. Over the years, inventories, reviews, and descriptions have been made. These contributions, apart from the taxonomic value, also provide valuable data on the occurrence of species and their distributions, which are rarely accompanied by notes about natural history and behavior. Keeping this legacy in mind, we collected dragonflies between 2011 and 2019 in Minas Gerais state, which resulted in the registration of 90 species, 41 genera and 11 families. Our results also increase distribution data, an important tool for conservation actions, and provide additional information about habitat and biology of species.

Resumo: O conhecimento sobre a riqueza e distribuição das libélulas do Brasil ainda está sendo desvelado. Ao longo dos anos, inventários, revisões e descrições foram feitas. Essas contribuições, além do valor taxonômico, também fornecem um conjunto de dados precioso sobre a ocorrência das espécies e suas distribuições, estas que raramente são acompanhadas de anotações sobre história natural e comportamento. Mantendo este legado em mente, nós realizamos coletas de libélulas entre 2011 e 2019 no estado de Minas Gerais, o que resultou no registro de 90 espécies, 41 gêneros e 11 famílias. Nossos resultados aumentam os dados de distribuição, uma ferramenta importante para ações de conservação, e fornecem informações sobre o habitat e a biologia das espécies.

Odonates from Bodoquena Plateau: checklist and information about endangered species / Libélulas da Serra da Bodoquena: lista de espécies e informações sobre espécies ameaçadas

Abstract Here we provide an updated checklist of the odonates from Bodoquena Plateau, Mato Grosso do Sul state, Brazil. We registered 111 species from the region. The families with the highest number of species were Libellulidae (50 species), Coenagrionidae (43 species) and Gomphidae (12 species). 35 species are registered in the IUCN Red List species, four being Data Deficient, 29 of Least Concern and two species being in the threatened category. Phyllogomphoides suspectus Belle, 1994 (Odonata: Gomphidae) was registered for the first time in the state.

Resumo Nós apresentamos um inventário atualizado das espécies de libélulas ocorrentes na Serra da Bodoquena, Estado de Mato Grosso do Sul, Brasil. Nós registramos 111 espécies para a região. As famílias com o maior número de espécies foram Libellulidae (50 espécies), Coenagrionidae (43 espécies) e Gomphidae (12 espécies). 35 espécies são registradas na Lista Vermelha de espécies da IUCN e duas estão em categorias de ameaçadas. Phyllogomphoides suspectus Belle, 1994 (Odonata: Gomphidae) foi registrado pela primeira vez no estado.