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Objective: Speckled Dace Rhinichthys osculus is small cyprinoid fish that is widespread in western North America. In California and elsewhere it is currently treated as a single species with multiple subspecies, many undescribed. However, these subspecies may represent evolutionary lineages that are cryptic species because they cannot be distinguished using standard morphometric techniques. In this study, we attempt to determine evolutionary lineages within California populations of Speckled Dace using the population genetic and genomic information. Methods: We used restriction site-associated DNA sequencing to extract thousands of single-nucleotide polymorphisms across the genome to identify genetic differences among all the samples from 38 locations in the western USA, with a focus on California. We performed principal component analysis, admixture analysis, estimated pairwise values of the genetic differentiation index FST, and constructed molecular phylogenies to characterize population genetic and phylogenetic relationships among sampled Speckled Dace populations. Result: Our analyses detected three major lineages of Speckled Dace in California that align with geography: (1) Sacramento River, central California coast, Klamath River, and Warner Basin; (2) Death Valley and Lahontan Basin; and (3) Santa Ana River basin, in southern California. These lineages fit well with the geologic history of California, which has promoted long isolation of populations of Speckled Dace and other fishes. Conclusion: The presence of distinct evolutionary lineages indicates that Speckled Dace in California should be managed with distinct population segments to preserve within-species diversity. This study highlights the importance of genetic analyses for conservation and management of freshwater fishes.

Pangolins are scaly and toothless mammals which are distributed across Africa and Asia. Currently, the Malayan, Chinese and Philippine pangolins are designated as critically endangered species. Although few pangolin viruses have been described, their viromes have received more attention following the discovery that they harbour sarbecoviruses related to SARS-CoV-2. Using large-scale genome mining, we discovered novel lineages of papillomaviruses infecting the Malayan and Chinese pangolins. We were able to assemble three complete circular papillomavirus genomes with an intact coding capacity and five additional L1 genes encoding the major capsid protein. Phylogenetic analysis revealed that seven out of eight L1 sequences formed a monophyletic group which is the sister lineage to the Tupaia belangeri papillomavirus 1, isolated from Yunnan province in China. Additionally, a single L1 sequence assembled from a Chinese pangolin was placed in a clade closer to Alphapapillomavirus and Omegapapillomavirus . Examination of the SRA data from 95 re-sequenced genomes revealed that 49.3% of Malayan pangolins and 50% of Chinese pangolins were positive for papillomavirus reads. Our results indicate that pangolins in South-East Asia are the hosts of diverse and highly prevalent papillomaviruses, and highlight the value of in silico mining of host sequencing data for the discovery of novel viruses.

This work was supported by ONR grants N000141613017 to E.L.C. and N.A. and N00014-18-1-2808 to C.S.B.; funds from the NMNH Rebecca G. Mead and James G. Mead Marine Mammal Endowment, NSF (USA) grant no. DEB-1457735 to M.S.S., P.A.M. and J.G.; Brothers Hartmann Foundation grant no. AB28148 to M.T.O.; NMFS, BOEM, and USA Navy funding to D.Ch. under the Atlantic Marine Assessment Program for Protected Species. M.L.M. was funded under the Marie Skłodowska-Curie grant agreement no 801199; E.L.C. by a Rutherford Discovery Fellowship from the Royal Society of New Zealand Te Apārangi. Irish Whale and Dolphin Group Cetacean Stranding scheme is part-funded by the National Parks and Wildlife Service. The deep sea has been described as the last major ecological frontier, as much of its biodiversity is yet to be discovered and described. Beaked whales (ziphiids) are among the most visible inhabitants of the deep sea, due to their large size and worldwide distribution, and their taxonomic diversity and much about their natural history remain poorly understood. We combine genomic and morphometric analyses to reveal a new Southern Hemisphere ziphiid species, Ramari's beaked whale, Mesoplodon eueu, whose name is linked to the Indigenous peoples of the lands from which the species holotype and paratypes were recovered. Mitogenome and ddRAD-derived phylogenies demonstrate reciprocally monophyletic divergence between M. eueu and True's beaked whale (M. mirus) from the North Atlantic, with which it was previously subsumed. Morphometric analyses of skulls also distinguish the two species. A time-calibrated mitogenome phylogeny and analysis of two nuclear genomes indicate divergence began circa 2 million years ago (Ma), with geneflow ceasing 0.35–0.55 Ma. This is an example of how deep sea biodiversity can be unravelled through increasing international collaboration and genome sequencing of archival specimens. Our consultation and involvement with Indigenous peoples offers a model for broadening the cultural scope of the scientific naming process. Publisher PDF

Among nudibranch molluscs, the family Tritoniidae gathers taxa with an uncertain phylogenetic position, such as some species of the genus Tritonia Cuvier, 1798. Currently, 37 valid species belong to this genus and only three of them are found in the Southern Ocean, namely T. challengeriana Bergh, 1884, T. dantarti Ballesteros & Avila, 2006, and T. vorax (Odhner, 1926). In this study, we shed light on the long-term discussed systematics and taxonomy of Antarctic Tritonia species using morpho-anatomical and molecular techniques. Samples from the Weddell Sea and Bouvet Island were dissected and prepared for scanning electron microscopy. The three molecular markers COI, 16S, and H3 were sequenced and analysed through maximum-likelihood and Bayesian methods. The phylogenetic analyses and species delimitation tests clearly distinguished two species, T. challengeriana widely spread in the Southern Ocean and T. dantarti endemic to Bouvet Island. Colouration seems to be an unreliable character to differentiate among species since molecular data revealed both species can either have orange or white colour morphotypes. This variability could be explained by pigment sequestration from the soft coral species they feed on. Morphological analyses reveal differences between Antarctic and Magellanic specimens of T. challengeriana. However, the relationship between T. challengeriana specimens from these two regions remains still unclear due to the lack of molecular data. Therefore, the validity of the T. antarctica Martens & Pfeffer, 1886, exclusively found in Antarctic waters requires further systematic work.

Specific status and species boundaries of several freshwater prawns in the Macrobrachium pilimanus species group remain ambiguous, despite the taxonomic re-description of type materials and additional specimens collected to expand the boundaries of some species. In this study, the “pilimanus” species group of Macrobrachium sensu Johnson (1958) was studied using specimens collected from montane streams of Thailand. Molecular phylogenetic analyses based on sequences of three molecular markers (COI, 16S and 18S rRNA) were performed. The phylogenetic results agreed with morphological identifications, and indicated the presence of at least nine putative taxa. Of these, six morphospecies were recognised as M. malayanum, M. forcipatum, M. dienbienphuense, M. hirsutimanus, M. eriocheirum, and M. sirindhorn. Furthermore, three morphologically and genetically distinct linages were detected, and are described herein as M. naiyanetri Siriwut sp. nov., M. palmopilosum Siriwut sp. nov. and M. puberimanus Siriwut sp. nov. The taxonomic comparison indicated wide morphological variation in several species and suggested additional diagnostic characters that are suitable for use in species diagnoses, such as the shape and orientation of fingers, the rostrum form, and the presence or absence of velvet pubescence hairs and tuberculated spinulation on each telopodite of the second pereiopods. The “pilimanus” species group was portrayed as non-monophyletic in both ML and BI analyses. The genetic structure of different geographical populations in Thailand was detected in some widespread species. The species delimitation based on the four delimitation methods (BIN, ABGD, PTP and GMYC) suggested high genetic diversity of the “pilimanus” species group and placed the candidate members much higher than in previous designations based on traditional morphology. This finding suggests that further investigation of morphological and genetic diversity of Southeast Asian freshwater prawns in the genus Macrobrachium is still required to provide a comprehensive species list to guide efforts in conservation and resource management.

By their past and present diversity, rodents are among the richest components of Caribbean land mammals. Many of these became extinct recently. Causes of their extirpation, their phylogenetic affinities, the timing of their arrival in the West Indies and their biogeographic history are all ongoing debated issues. Here, we report the discovery of dental remains from Lower Oligocene deposits (ca29.5 Ma) of Puerto Rico. Their morphology attests to the presence of two distinct species of chinchilloid caviomorphs, closely related to dinomyids in a phylogenetic analysis, and thus of undisputable South American origin. These fossils represent the earliest Caribbean rodents known thus far. They could extend back to 30 Ma the lineages of some recently extinct Caribbean giant rodents (ElasmodontomysandAmblyrhiza), which are also retrieved here as chinchilloids. This new find has substantial biogeographic implications because it demonstrates an early dispersal of land mammals from South America to the West Indies, perhaps via the emergence of the Aves Ridge that occurredca35–33 Ma (GAARlandia hypothesis). Considering both this new palaeontological evidence and recent molecular divergence estimates, the natural colonization of the West Indies by rodents probably occurred through multiple and time-staggered dispersal events (chinchilloids, then echimyid octodontoids (spiny rats/hutias), caviids and lastly oryzomyin muroids (rice rats)).

16S rRNA gene amplicon sequencing is a state of the art technology to analyze bacterial communities via microbiome profiling. Choosing an appropriate DNA extraction protocol is crucial for characterizing the microbial community and can be challenging, especially when preliminary knowledge about the sample matrix is scarce. The aim of the present study was to evaluate seven commercial DNA extraction kits suitable for 16S rRNA gene amplicon sequencing of the bacterial community of the chicken cecum, taking into account different criteria such as high technical reproducibility, high bacterial diversity and easy handling. The DNA extraction kits differed strongly with respect to extractable DNA quantity, DNA quality, technical reproducibility and bacterial diversity determined after 16S rRNA gene amplicon sequencing and subsequent bioinformatic and biostatistical data processing. While some of the DNA extraction protocols under-represented specific bacterial community members, the removal of PCR inhibitors supported technical reproducibility and subsequently enhanced the recovered bacterial diversity from the chicken cecum community. In conclusion, the removal of PCR inhibitors from the sample matrix seemed to be one of the main drivers for a consistent representation of the bacterial community even of low abundant taxa in chicken cecum samples., An accurate description of the broiler caecal microbiota is a challenging tasks starting from a suitable DNA extraction to get insights into this complex habitat that affects animal health.

Papillomaviruses (PVs) are ancient viruses infecting vertebrates, from fishes to mammals. Although the genomes of PVs are small and show conserved synteny, PVs display large genotypic diversity and ample variation in the phenotypic presentation of the infection. Most PV genomes contain two small early genes E6 and E7 . In a bunch of closely related human papillomaviruses (HPVs), the E6 and E7 proteins provide the viruses with oncogenic potential. The recent discoveries of PVs without E6 and E7 in different fish species place a new root on the PV tree, and suggest that ancestral PVs consisted of the minimal PV backbone E1-E2-L2-L1 . Bayesian phylogenetic analyses date the most recent common ancestor of the PV backbone to 424 million years ago (Ma). Common ancestry tests on extant E6 and E7 genes indicate that they share a common ancestor dating back to at least 184 Ma. In AlphaPVs infecting Old World monkeys and apes, the appearance of the E5 oncogene 53–58 Ma concurred with (i) a significant increase in substitution rate, (ii) a basal radiation and (iii) key gain of functions in E6 and E7. This series of events was instrumental to construct the extant phenotype of oncogenic HPVs. Our results assemble the current knowledge on PV diversity and present an ancient evolutionary timeline punctuated by evolutionary innovations in the history of this successful viral family. This article is part of the theme issue ‘Silent cancer agents: multi-disciplinary modelling of human DNA oncoviruses’.

International audience; Past global change may have forced animal-dispersed plants with megafaunal fruits to adapt or go extinct, but these processes have remained unexplored at broad spatio-temporal scales. Here, we combine phylogenetic, distributional and fruit size data for more than 2500 palm (Arecaceae) species in a time-slice diversification analysis to quantify how extinction and adaptation have changed over deep time. Our results indicate that extinction rates of palms with megafaunal fruits have increased in the New World since the onset of the Quaternary (2.6 million years ago). In contrast, Old World palms show a Quaternary increase in transition rates towards evolving small fruits from megafaunal fruits. We suggest that Quaternary climate oscillations and concurrent habitat fragmentation and defaunation of megafaunal frugivores in the New World have reduced seed dispersal distances and geographical ranges of palms with megafaunal fruits, resulting in their extinction. The increasing adaptation to smaller fruits in the Old World could reflect selection for seed dispersal by ocean-crossing frugivores (e.g. medium-sized birds and bats) to colonize Indo-Pacific islands against a background of Quaternary sea-level fluctuations. Our macro-evolutionary results suggest that megafaunal fruits are increasingly being lost from tropical ecosystems, either due to extinctions or by adapting to smaller fruit sizes.

Two different nematode species were recovered from pomegranate decaying fruit in two localities in Southern Italy: the mycetophagus nematode Sheraphelenchus sucus and a bacterial feeder nematode belonging to the Panagrolaimidae (Rhabditida) family. Morphometrics of the Italian population of S. sucus closely resemble that of the type population, whereas some differences were found when compared with another population from Iran. Molecular characterization of the Italian S. sucus using the 18S rRNA gene, D2–D3 expansion domains of the 28S rDNA, the ITS region, and the partial mitochondrial COI were carried out. Sequences of the 18S rRNA gene, the D2–D3 domains, and the ITS were analyzed using several methods for inferring phylogeny to reconstruct the relationships among Sheraphelenchus and Bursaphelenchus species. The bacterial feeder Panagrellus sp. was characterized at the molecular level only. The D2–D3 expansion domains and ITS sequences of this Italian panagrolaimid were determined. The D2–D3 sequences of the Italian panagrolaimid showed 99% similarity with the corresponding sequence of Panagrellus sp. associated with Rhynchophorus ferrugineus. This is the first report on the tritrophic association of S. sucus and Rhabditida that uses both insects and pomegranate fruit as hosts.

The taxonomy and the systematics of the genus Makatinus are discussed by means of the characterization of its morphological pattern and the first molecular (D2–D3 expansion segments of 28S rDNA) analysis of a representative of this taxon, Makatinus crassiformis from Costa Rica. The presence of two or more pairs of male ad-cloacal genital papillae is the most characteristic autapomorphy of the genus, but the status of its species on this concern differ among them. Both morphological and molecular data support a relationship with Aporcelaimellus, which, however, might not be as close as usually assumed. An emended diagnosis of the genus, a key to species identification, and a compendium of their morphometrics are provided. Makatinus siddiqii is regarded as species inquirenda, Makatinus simus is retained under Eudorylaimus, and Makatinus tritici becomes a junior synonym of Aporcelaimellus tritici.

BackgroundThe corallum is crucial in building coral reefs and in diagnosing systematic relationships in the order Scleractinia. However, molecular phylogenetic analyses revealed a paraphyly in a majority of traditional families and genera among Scleractinia showing that other biological attributes of the coral, such as polyp morphology and reproductive traits, are underutilized. Among scleractinian genera, theEuphyllia, with nine nominal species in the Indo-Pacific region, is one of the groups that await phylogenetic resolution. Multiple genetic markers were used to construct the phylogeny of sixEuphylliaspecies, namelyE. ancora, E. divisa, E. glabrescens, E. paraancora, E. paradivisa,andE. yaeyamaensis.The phylogeny guided the inferences on the contributions of the colony structure, polyp morphology, and life history traits to the systematics of the largest genus in Euphyllidae (clade V) and, by extension, to the rest of clade V.ResultsAnalyses of cytochrome oxidase 1 (cox1), cytochrome b (cytb), and β-tubulin genes of 36 colonies representingEuphylliaand a confamilial species,Galaxea fascicularis,reveal two distinct groups in theEuphylliathat originated from different ancestors.Euphyllia glabrescensformed a separate group.Euphyllia ancora, E. divisa, E. paraancora, E. paradivisa,andE. yaeyamaensisclustered together and diverged from the same ancestor asG. fascicularis.The 3′-end of thecox1gene ofEuphylliawas able to distinguish morphospecies.DiscussionSpecies ofEuphylliawere traditionally classified into two subgenera,EuphylliaandFimbriaphyllia,which represented a dichotomy on colony structure. The paraphyletic groups retained the original members of the subgenera providing a strong basis for recognizingFimbriaphylliaas a genus. However, colony structure was found to be a convergent trait betweenEuphylliaandFimbriaphyllia,while polyp shape and length, sexuality, and reproductive mode defined the dichotomy better. Species in a genus are distinguished by combining polyp morphology and colony form. The cluster ofE. glabrescensof theEuphylliagroup is a hermaphroditic brooder with long, tubular tentacles with knob-like tips, and a phaceloid colony structure. TheFimbriaphylliagroup, withF. paraancora, F. paradivisa, F. ancora, F. divisa,andF. yaeyamaensis,are gonochoric broadcast spawners with short polyps, mixed types of tentacle shapes, and a phaceloid or flabello-meandroid skeleton. Soft-tissue morphology ofG. fascicularisandCtenella chagiuswere found to be consistent with the dichotomy.ConclusionsThe paraphyly of the original members of the previous subgenera justify recognizingFimbriaphylliaas a genus. The integrated approach demonstrates that combining polyp features, reproductive traits, and skeletal morphology is of high systematic value not just toEuphylliaandFimbriaphylliabut also to clade V; thus, laying the groundwork for resolving the phylogeny of clade V.

In the nematode family Criconematidae, a taxonomy primarily based on cuticle characters has created classifications that are notoriously volatile. Molecular characters may lead to their stabilization. A phylogenetic tree of Criconematoidea was constructed using 166 new near full-length 18S rDNA sequences and 58 sequences from GenBank. Bayesian and maximum likelihood (ML) analyses produced trees with similar topologies. Major features include a strongly supported clade that includes Criconematidae and Hemicycliophoridae, excluding Paratylenchidae and Tylenchulidae. Another well-supported clade groups Criconema, Ogma, Crossonema, and Hemicriconemoides plus Xenocriconemella, combining nematodes with cuticular scales with those without scales at any life stage. Mesocriconema, Discocriconemella limitanea, Hemicaloosia, and Lobocriconema are recognized as monophyletic groups, but Criconemoides is paraphyletic. Both trees support an unexpected sister relationship between Bakernema and Hemicycliophora. The 18S rDNA dataset was insufficient for distinguishing genus boundaries between Criconema, Ogma, and Crossonema. The relationships depicted by the 18S rDNA phylogeny suggest that key morphological characters used in the classification of Criconematidae are not homologous.

Pratylenchus haiduongensis sp. n. is described as associated with carrot (Daucus carota subsp. sativus (Hoffm.) Schübl. & G. Martens) in Hai Duong Province, Vietnam. P. haiduongensis sp. n. is characterized by the lip region with three annuli and slightly separated from the body. Stylet knobs are rounded (never indented anteriorly). The lateral field includes four incisures, bearing areolation at the pharynx region and tail region and occasionally appears in the vulval region. Sometimes the appearances of oblique broken striaes divide the lateral field into five or six incisures. The ovary is distinct with one row of oocytes. Spermatheca is oval in shape with round central cavity, without sperm or reduced in some specimens. The postvuval uterine sac is long surpassing the vulva body diameter by 2 to 2.5 times (PUS = 31 to 65 µm). High vulva position with V = 66 to 75%. The tail shape can be subhemispherical with a smooth, slightly indented, broadly smooth, or cleft terminus observed in some specimens. The matrix code of P. haiduongensis sp. n. is: A2, B1, C4, D(1,3), E1, F(5,6), G(1,2), H(1,4); I(1,2,3,4), J1, K(1,2) according to Castillo and Vovlas (2007). The LSU–D2D3 segment and the ITS–rDNA region of this species were amplified and sequenced. The morphological characters and molecular phylogenetic analyses confirmed that this is a new species of the genus Pratylenchus in Vietnam.

Anguillonema amolensis n. sp. is described and illustrated based on its morphological, morphometric, and molecular characters. The new species is characterized by its 575 to 820 μm long and wide body (body width at vulva = 30 to 59 μm), irregularly ventrally curved after fixation, five to six lines in lateral fields, 6.0 to 7.5 μm long stylet with small rounded knobs, pharynx lacking a median bulb, pharyngo-intestinal junction anterior to nerve ring and excretory pore, females with monodelphic-prodelphic reproductive system, 15 to 19 μm long conical tail with broad rounded tip, and males absent. The new species is compared with two known species of the genus, Anguillonema poligraphi and A. crenati. Molecular phylogenetic studies of the new species using partial sequences of small subunit (SSU) rDNA revealed that it forms a clade with an unidentified nematode species and two species of the genus Howardula. In phylogenetic analyses using partial sequences of the 28S rDNA (D2-D3 segment), the new species formed a monophyletic group with species belonging to two genera Howardula and Parasitylenchus.

Xiphinema japonicum n. sp., isolated in Ningbo, China, from the rhizosphere of Podocarpus macrophyllus (Thunb.) imported from Japan is described. The new species belongs to Xiphinema non-americanum group 7 and is characterized by medium body length (3.0–3.7 mm), total stylet length 190–201 μm, vulva located anteriorly (V = 30.5%–35.3%), two equally developed female genital branches without uterine differentiation (no Z or pseudo-Z organ and/or spines in the uteri), short tail, convex-conoid with subdigitate peg in terminus, and absence of males. The species has four juvenile developmental stages (J1 was not found). The polytomous identification codes of the new species are (codes in parentheses are exceptions) A4-B4-C4-D5(4)-E2(3)-F3(4)-G2(3)-H2-I3-J4-K?-L1. Morphologically, the new species is mainly characterized by combination of the codes C4 and E2(3), making the species unique and different from other species in the genus. It is most similar to the North American species Xiphinema bakeri, herein considered as its cryptic species by the nature of high morphological similarity, but with significant differences in DNA sequences in nearly full length 18S, ITS1, 28S D2/D3, and cytochrome c oxidase subunit 1 sequences. The second morphologically close species is Xiphinema setariae from which the new species could be separated based on morphological and molecular characters. Morphological comparisons with phylogenetically related species are also discussed.

Nothotylenchus phoenixae n. sp. is described and illustrated from soil samples of palm trees in Kermanshah Province, western Iran. The new species is characterized by a body length of 784 (663 to 925) mm in females and 677 to 715 mm in males; a delicate stylet 6 (5 to 7) mm long and six lines in the lateral field; median bulb of pharynx fusiform, nonmuscular, and nonvalvate; isthmus elongate, slender ending to a pyriform basal pharyngeal bulb not overlapping intestine; postvulval uterine sac well developed, 15 (14 to 17) mm long, female tail elongate-conoid with pointed terminus; and male with adanal bursa and spicules 21 to 22 mm long (n = 2). The new species comes close in morphology and morphometrics to five known species of the genus, namely N. affinis, N. hexaglyphus, N. persicus, N. taylori, and N. uniformis. Molecular analyses of the partial 18S, D2/D3 expansion segments of the partial 28S and internal transcribed spacer (ITS) revealed this as a new species. The sequences of the partial 18S and 28S D2/D3 regions confirmed the close phylogenetic relationship between N. phoenixae n. sp. and other anguinids, but Nothotylenchus is clearly separated from Ditylenchus species and should be considered as a valid genus.

Fourteen populations of Meloidogyne graminicola were collected from different agroecological regions of India. Morphological and morphometrical comparisons were made for various nematode life stages. Three populations (Hisar, New Delhi, and Samastipur) were different from typical M. graminicola on the basis of the length of eggs; J2 length, a-value, hyaline tail portion; male length, distance up to excretory pore, spicule and gubernaculum lengths; female length and width, stylet length, distance up to excretory pore, EPST (distance of excretory pore from anterior end / stylet length [females]) ratio, and vulval length. Morphological and morphometrical comparison with closely related species M. graminis, M. oryzae, M. salasi, M. triticoryzae, and M. lini clustered these populations into two groups: Anand, Bhubaneswar, Hyderabad, Jammu, Jorhat, Kalyani, Kanpur, Ludhiana, Mandya, Palampur, Vellayani grouped with M. graminicola, M. triticoryzae and M. salasi; whereas, Hisar, New Delhi, Samastipur grouped with M. oryzae and M. graminis. Molecular phylogenetic analysis using internal transcribed spacer (ITS) suggested that in spite of morphological differences, these populations belonged to M. graminicola.

Our recent study evaluated the performance of parsimony and probabilistic models of phylogenetic inference based on categorical data [1]. We found that a Bayesian implementation of a probabilistic Markov model produced more accurate results than either of the competing parsimony approaches (the main method currently employed), and the maximum-likelihood implementation of the same model. This occurs principally because the results of Bayesian analyses are less resolved (less precise) as a measure of topological uncertainty is intrinsically recovered in this MCMC-based approach and can be used to construct a majority-rule consensus tree that reflects this. Of the three main methods, maximum likelihood performed theworst of all as a single exclusively bifurcating tree is estimated in this framework which does not integrate an intrinsic measure of support.

Molecular biology has provided a rich dataset to develop hypotheses of nervous system evolution. The startling patterning similarities between distantly related animals during the development of their central nervous system (CNS) have resulted in the hypothesis that a CNS with a single centralized medullary cord and a partitioned brain is homologous across bilaterians. However, the ability to precisely reconstruct ancestral neural architectures from molecular genetic information requires that these gene networks specifically map with particular neural anatomies. A growing body of literature representing the development of a wider range of metazoan neural architectures demonstrates that patterning gene network complexity is maintained in animals with more modest levels of neural complexity. Furthermore, a robust phylogenetic framework that provides the basis for testing the congruence of these homology hypotheses has been lacking since the advent of the field of ‘evo-devo’. Recent progress in molecular phylogenetics is refining the necessary framework to test previous homology statements that span large evolutionary distances. In this review, we describe recent advances in animal phylogeny and exemplify for two neural characters—the partitioned brain of arthropods and the ventral centralized nerve cords of annelids—a test for congruence using this framework. The sequential sister taxa at the base of Ecdysozoa and Spiralia comprise small, interstitial groups. This topology is not consistent with the hypothesis of homology of tripartitioned brain of arthropods and vertebrates as well as the ventral arthropod and rope-like ladder nervous system of annelids. There can be exquisite conservation of gene regulatory networks between distantly related groups with contrasting levels of nervous system centralization and complexity. Consequently, the utility of molecular characters to reconstruct ancestral neural organization in deep time is limited.

Knowledge of absolute species divergence times is not only fascinating to evolutionary biologists in establishing the age of a species group, but also critically important to addressing a variety of biological questions. Absolute times allow us to place speciation events (such as the diversification of the mammals relative to the demise of the dinosaurs) in the correct geological and environmental contexts and to gain a better understanding of speciation and dispersal mechanisms [1,2]. They also allow us to characterize species richness and species diversification rates over geological periods. Estimated molecular evolutionary rates can also be correlated with life-history traits and are important for interpretation of the fast-accumulating genomic sequence data. Molecular clock methods are also used widely in establishing the evolutionary history of viruses, including those related to human diseases.

In a survey of ecotypes for longidorids, primarily from the rhizosphere hardwood trees growing in sandy soil along stream banks, 828 soil samples were collected from 37 Arkansas counties in 1999-2001. Eight populations of Longidorus breviannulatus were identified from the Arkansas survey samples. A total of 19 populations from California, Illinois, Iowa, Kansas, Michigan, Nebraska, New Jersey, New York, and Wisconsin were identified from the collection of the second author. A few males were found in New York and Nebraska populations and are described herein. Seven populations of L. fragilis were identified in the Arkansas survey samples, and one population was found from Indiana. Four juvenile stages of L. fragilis are present, and data are given for them herein.

Longidorus crassus is a common species and widely distributed in Arkansas. It was also identified for the first time in samples from Alabama, Iowa, Kansas, Nebraska, South Carolina, Wisconsin, and Canada. It is a parthenogenetic species, but a few males were found and were described herein for the first time. Four developmental juvenile stages were identified. A high degree of intraspecies variation was observed among different populations of this species. Twenty-three populations of L. crassus found in Arkansas were studied for their variability using standard measurements, mean comparison, and coefficient of variation. Most of the Arkansas populations have a smaller body than the paralectotypes. Populations Long-63 and Long-88 are close to the paralectotype population. Two populations, Long-10 and Long-80, are different from each other and all other populations. The majority of morphometric characters of this species do not have a normal distribution pattern as they have a high degree of variability within and between populations. The means of many morphometric characters strikingly differ between populations. Hierarchical cluster analysis based on female morphometric character means including body length, distance from vulva opening to anterior end, head width, odontostyle length, esophagus length, body width, tail length, and anal body width were used to examine the morphometric relationships and create dendrograms for 23 Arkansas populations and the lectotype population.

Hierarchical cluster analysis based on female morphometric character means including body length, distance from vulva opening to anterior end, head width, odontostyle length, esophagus length, body width, tail length, and tail width were used to examine the morphometric relationships and create dendrograms for (i) 62 populations belonging to 9 Longidorus species from Arkansas, (ii) 137 published Longidorus species, and (iii) 137 published Longidorus species plus 86 populations of 16 Longidorus species from Arkansas and various other locations by using JMP 4.02 software (SAS Institute, Cary, NC). Cluster analysis dendograms visually illustrated the grouping and morphometric relationships of the species and populations. It provided a computerized statistical approach to assist by helping to identify and distinguish species, by indicating morphometric relationships among species, and by assisting with new species diagnosis. The preliminary species identification can be accomplished by running cluster analysis for unknown species together with the data matrix of known published Longidorus species.