Palaearctic species of the ichneumon-fly genus Campodorus Förster (Hymenoptera, Ichneumonidae): III. Species with long-haired ovipositor sheath, species with entirely rufous hind tibia, and species with white-banded tibiae

A complex of about 40 Palaearctic species of the genus Campodorus with long haired ovipositor sheath and with entirely rufous or white-banded hind tibia is considered and keyed. Twenty-one new species are described. The species with the pale rufous hind tibia have black hind coxa and trochanters, most of them also have rather long genae and scabrous mesopleura. The majority of these species are distributed in subarctic Siberia: C. yakutator sp. n., C. minutor sp. n., C. arctor sp. n., C. atripes sp. n., C. pequenitor sp. n., C. mordax sp. n., C. lucidator sp. n., C. subarctor sp. n., C. nikandrovskii sp. n., and C. genator sp. n. C. suomi sp. n. is described from northern Finland, C. labytnangi sp. n., from Kola Peninsula, the Polar Ural, and Transbaikalia, and only C. gallicator sp. n., from France (? Alps). The species with a white band on the hind tibia are mostly boreal; most of them have red coxae and trochanters (except for C. sakhalinator sp. n. and C. taigator sp. n.), rather short genae, and granulate mesopleura. In this group, eight new species are described: C. atrofemorator sp. n., C. barbator sp. n., C. sakhalinator sp. n. (all from the southern part of the Russian Far East), C. taigator sp. n. (southern Siberian), C. ciliator sp. n., C. melanopygus sp. n., C. marginator sp. n., and C. boreator sp. n. (European-Siberian).     

In vitro activity effects of combinations of cephalothin, dicloxacillin, imipenem, vancomycin and amikacin against methicillin-resistantStaphylococcus spp. strains

Background

CHROMagar Candida (CaC) is increasingly being reported as a medium used to differentiate Candida albicans from non-albicans Candida (NAC) species. Rapid identification of NAC can assist the clinician in selecting appropriate antifungal therapy. CaC is a differential chromogenic medium designed to identify C. albicans, C. krusei, and C. tropicalis based on colony color and morphology. Some reports have proposed that CaC can also reliably identify C. dubliniensis and C. glabrata.

Methods

We evaluated the usefulness of CaC in the identification of C. dubliniensis, C. famata, C. firmetaria, C. glabrata, C. guilliermondii, C. inconspicua, C. kefyr, C. lipolytica, C. lusitaniae, C. norvegensis, C. parapsilosis, and C. rugosa.

Results

Most NAC produced colonies that were shades of pink, lavender, or ivory. Several isolates of C. firmetaria and all C. inconspicua produced colonies difficult to differentiate from C. krusei. Most C. rugosa isolates produced unique colonies with morphology like C. krusei except in a light blue-green color. C. glabrata isolates produced small dark violet colonies that could be differentiated from the pink and lavender colors produced by other species. All seventeen isolates of C. dubliniensis produced green colonies similar to those produced by C. albicans.

Conclusion

C. glabrata and C. rugosa appear distinguishable from other species using CaC. Some NAC, including C. firmetaria and C. inconspicua, could be confused with C. krusei using this medium.     

Comparative mitochondrial genomics and phylogenetic relationships of the Crossoptilon species (Phasianidae,Galliformes)

Background

Phasianidae is a family of Galliformes containing 38 genera and approximately 138 species, which is grouped into two tribes based on their morphological features, the Pheasants and Partridges. Several studies have attempted to reconstruct the phylogenetic relationships of the Phasianidae, but many questions still remain unaddressed, such as the taxonomic status and phylogenetic relationships among Crossoptilon species. The mitochondrial genome (mitogenome) has been extensively used to infer avian genetic diversification with reasonable resolution. Here, we sequenced the entire mitogenomes of three Crossoptilon species (C. harmani, C. mantchuricum and C. crossoptilon) to investigate their evolutionary relationship among Crossoptilon species.

Results

The complete mitogenomes of C. harmani, C. mantchuricum and C. crossoptilon are 16682 bp, 16690 bp and 16680 bp in length, respectively, encoding a standard set of 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a putative control region. C. auritum and C. mantchuricum are more closely related genetically, whereas C. harmani is more closely related to C. crossoptilon. Crossoptilon has a closer relationship with Lophura, and the following phylogenetic relationship was reconstructed: ((Crossoptilon + Lophura) + (Phasianus + Chrysolophus)). The divergence time between the clades C. harmani-C. crossoptilon and C. mantchuricum-C. auritum is consistent with the uplift of the Tibetan Plateau during the Tertiary Pliocene. The Ka/Ks analysis showed that atp8 gene in the Crossoptilon likely experienced a strong selective pressure in adaptation to the plateau environment.

Conclusions

C. auritum with C. mantchuricum and C. harmani with C. crossoptilon form two pairs of sister groups. The genetic distance between C. harmani and C. crossoptilon is far less than the interspecific distance and is close to the intraspecific distance of Crossoptilon, indicating that C. harmani is much more closely related to C. crossoptilon. Our mito-phylogenomic analysis supports the monophyly of Crossoptilon and its closer relationship with Lophura. The uplift of Tibetan Plateau is suggested to impact the divergence between C. harmani-C. crossoptilon clade and C. mantchuricum-C. auritum clade during the Tertiary Pliocene. Atp8 gene in the Crossoptilon species might have experienced a strong selective pressure for adaptation to the plateau environment.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1234-9) contains supplementary material, which is available to authorized users.     

Multi Locus Sequence Typing of Chlamydia Reveals an Association between Chlamydia psittaci Genotypes and Host Species

Chlamydia comprises a group of obligate intracellular bacterial parasites responsible for a variety of diseases in humans and animals, including several zoonoses. Chlamydia trachomatis causes diseases such as trachoma, urogenital infection and lymphogranuloma venereum with severe morbidity. Chlamydia pneumoniae is a common cause of community-acquired respiratory tract infections. Chlamydia psittaci, causing zoonotic pneumonia in humans, is usually hosted by birds, while Chlamydia abortus, causing abortion and fetal death in mammals, including humans, is mainly hosted by goats and sheep. We used multi-locus sequence typing to asses the population structure of Chlamydia. In total, 132 Chlamydia isolates were analyzed, including 60 C. trachomatis, 18 C. pneumoniae, 16 C. abortus, 34 C. psittaci and one of each of C. pecorum, C. caviae, C. muridarum and C. felis. Cluster analyses utilizing the Neighbour-Joining algorithm with the maximum composite likelihood model of concatenated sequences of 7 housekeeping fragments showed that C. psittaci 84/2334 isolated from a parrot grouped together with the C. abortus isolates from goats and sheep. Cluster analyses of the individual alleles showed that in all instances C. psittaci 84/2334 formed one group with C. abortus. Moving 84/2334 from the C. psittaci group to the C. abortus group resulted in a significant increase in the number of fixed differences and elimination of the number of shared mutations between C. psittaci and C. abortus. C. psittaci M56 from a muskrat branched separately from the main group of C. psittaci isolates. C. psittaci genotypes appeared to be associated with host species. The phylogentic tree of C. psittaci did not follow that of its host bird species, suggesting host species jumps. In conclusion, we report for the first time an association between C. psittaci genotypes with host species.     

Extreme diversification driven by parallel events of massive loss of heterozygosity in the hybrid lineage of Candida albicans

Candida albicans is the most commonly reported species causing candidiasis. The taxonomic classification of C. albicans and related lineages is controversial, with Candida africana (syn. C. albicans var. africana) and Candida stellatoidea (syn. C. albicans var. stellatoidea) being considered different species or C. albicans varieties depending on the authors. Moreover, recent genomic analyses have suggested a shared hybrid origin of C. albicans and C. africana, but the potential parental lineages remain unidentified. Although the genomes of C. albicans and C. africana have been extensively studied, the genome of C. stellatoidea has not been sequenced so far. In order to get a better understanding of the evolution of the C. albicans clade, and to assess whether C. stellatoidea could represent one of the unknown C. albicans parental lineages, we sequenced C. stellatoidea type strain (CBS 1905). This genome was compared to that of C. albicans and of the closely related lineage C. africana. Our results show that, similarly to C. africana, C. stellatoidea descends from the same hybrid ancestor as other C. albicans strains and that it has undergone a parallel massive loss of heterozygosity.     

Study on Cortinarius subgenus Telamonia section Hydrocybe in Europe, with especial emphasis on Mediterranean taxa

In this paper we have attempted to clarify the taxonomy and nomenclature of thirteen taxa of the genus Cortinarius subgenus Telamonia (sections Hydrocybe, Fraternii) well represented in the southwestern Mediterranean area of Europe (C. atrocoeruleus, C. bombycinus, C casimiri, C. contrarius, C. decipiens, C. fraternus, C. gallurae, C. hoffmannii, C. petroselineus, C. sertipes, C. subturibulosus, C. urdaibaiensis and C. vernus). To this end we have performed a combined study of morphological and molecular data (rDNA ITS sequences). The morphological analysis was carried out on 114 collections and the molecular analysis involved 31 of the 114 collections, including 11 type collections (types for C. casimiri and C. fraternus were not available). In addition, a study of spores under field emission scanning electron microscopy (FESEM) was conducted. The results of the combined analysis allowed us to asign the studied material to five species (C. casimiri s.l., C. decipiens s.l., C. gallurae, C. subturibulosus s.l. and C. vernus s.l.). Thus, all collections from more continental areas, which were originally identified as six different taxa (C. atrocoeruleus, C. contrarius, C. decipiens, C. fraternus, C. sertipes, C. flexipes fo. sertipes) corresponded to C. decipiens sensu lato, a widely distributed, genetically and morphologically variable species. Cortinarius casimiri is also found in such habitats, but it is confirmed as distinct taxon. Collections from Mediterranean sclerophyllous communities correspond to C. gallurae, C. vernus sensu lato and C. subturibulosus sensu lato. Due to close phylogenetic relationships we propose the new combinations C. casimiri var. hoffmannii (=C. decipiens var. hoffmannii non C. hoffmannii) and C. subturibulosus var. bombycinus (=C. bombycinus), and the new variety C. vernus var. nevadavernus (=C. vernus H. Lindstr. & Melot sensu auct.).     

Systematics of Crepidostomum species from the Russian Far East and northern Japan,with description of a new species and validation of the genus Stephanophiala

Current article touched upon the issue of the complicated taxonomic status of some species from the genus Crepidostomum collected from the freshwater fish in the rivers of Primorsky region, Sakhalin, and Hokkaido Islands. Primary morphological analyses showed affiliation of the worms to the species C. farionis (Müller, 1784) Lühe, 1909; C. metoecus Braun, 1900b; C. chaenogobii Yamaguti and Matsumura, 1942; C. nemachilus Krotov, 1959. We described the new species Crepidostomum achmerovi sp. nov. that is a sibling species of C. nemachilus. Molecular-genetic investigation have shown that C. nemachilus and C. achmerovi sp. nov. are closely related to C. metoecus in both 28S rDNA and cox1 mtDNA markers. Crepidostomum nemachilus forms a separate branch within the C. metoecus clade on the 28S BI tree with strong statistical support and separate clade in relation to C. metoecus clade on the cox1 BI tree. Values of p-distances between Crepidostomum species were at intergeneric level. Crepidostomum metoecus species complex including five species (C. metoecus, C. nemachilus, C. oschmarini, C. brinkmanni, and C. achmerovi sp. nov.) was reconsidered as independent genus Crepidostomum sensu stricto. Minimum Spanning Network showed that C. nemachilus, C. metoecus and C. achmerovi sp. nov. were separated by large number of mutational events and represent independent phyletic lines. An amended diagnosis is provided for the subfamily Crepidostomatinae, the genera Crepidostomum s. str. and Stephanophiala Nicoll, 1909, along with keys to species of both genera.     

Ecological survey of two Calomyscidae species; Goodwin's brush-tailed mouse and Hotson's brush-tailed mouse (Rodentia) in the eastern parts of Iran

Calomyscus elburzensis and C. hotsoni are two species of the rodents family Calomyscidae which are distributed mainly in Iran. Herein, we evaluated the habitat and ecological differences of these two brush-tailed mice in order to test the credibility of a hypothesis stating that species from habitats with different climates and vegetation show greater intraspecific differentiation than those from areas with more similar climates and vegetation. This study was carried out in four rocky regions in Iran between 2013 and 2015. Totally 52 brush-tailed mice were captured from Kopet-Dag, Khaje-Morad, Ark, and Shadan and Olang during the field studies. Maximum parsimony analysis inferred from mitochondrial DNA sequences (Cytb) was used for species identification, and also comparison of mean Kimura 2-paramater distances was performed. According to the molecular studies, specimens from the first two regions were assigned to C. elburzensis and samples from Ark, and Shadan and Olang belonged to C. hotsoni. The mean distances within all examined Iranian samples of both C. elburzensis and C. hotsoni were 2.3% and 0.9%, respectively. Based on our field studies, C. elburzensis were captured either from “cold mountainous” climate zone with Juniperus excelsa as main vegetation cover or from “Mediterranean” in which Pistacia atlantica is predominant vegetation. C. hotsoni were found in “hot dry desert” and “cold semi-desert” regions both characterized by Ephedra sp. and Avena sativa vegetation cover. Dog rose seeds were the main food of C. elburzensis in Kopet-Dag, whereas brush-tailed mice in the other three regions fed mainly on Mount Atlas pistache fruits. C. elburzensis inhabited concealed rock crevices, but C. hotsoni were found living in burrows which were digged in the soft soils. Individuals of C. hotsoni showed more calm behavior as compared with C. elburzensis. Meriones persicus and Cricetulus migratorius (Rodentia) as well as Ochotona rufescence (Lagomorpha) were the most abundant small mammals encountered with C. elburzensis, while M. persicus was the most successful rodent with sympatric colonies with C. hotsoni. In conclusion, in accordance with the greater genetic distances observed in C. elburzensis populations as compared with C. hotsoni, more significant differences in the habitat structure were also found for the first species in different parts of its distribution range.     

Molecular characterization and morphology of Cochlodinium strangulatum,the type species of Cochlodinium,and Margalefidinium gen. nov. for C. polykrikoides and allied species (Gymnodiniales,Dinophyceae)

Photosynthetic species of the dinoflagellate genus Cochlodinium such as C. polykrikoides, one of the most harmful bloom-forming dinoflagellates, have been extensively investigated. Little is known about the heterotrophic forms of Cochlodinium, such as its type species, Cochlodinium strangulatum. This is an uncommon, large (∼200 μm long), solitary, and phagotrophic species, with numerous refractile bodies, a central nucleus enclosed in a distinct perinuclear capsule, and a cell surface with fine longitudinal striae and a circular apical groove. The morphology of C. polykrikoides and allied species is different from the generic type. It is a bloom-forming species with single, two or four-celled chains, small cell size (25–40 μm long) with elongated chloroplasts arranged longitudinally and in parallel, anterior nucleus, eye-spot in the anterior dorsal side, and a cell surface smooth with U-shaped apical groove. Phylogenetic analysis based on LSU rDNA sequences revealed that C. strangulatum and C. polykrikoides/C. fulvescens formed two distally related, independent lineages. Based on morphological and phylogenetic analyses, the diagnosis of Cochlodinium is emended and C. miniatum is proposed as synonym of C. strangulatum. The new genus Margalefidinium gen. nov., and new combinations for C. catenatum, C. citron, C. flavum, C. fulvescens and C. polykrikoides are proposed.     

The influence of pre- and post-zygotic barriers on interspecific Corymbia hybridization

Background and Aims

Corymbia species from different sections hybridize readily, with some of increasing economic importance to plantation forestry. This study explores the locations of reproductive barriers between interspecific Corymbia hybrids and investigates the reproductive success of a wide taxonomic range of C. torelliana hybrid crosses.

Methods

Pollen, pistil and embryo development were investigated for four C. torelliana crosses (×C. torelliana, ×C. citriodora subsp. citriodora, ×C. tessellaris and ×C. intermedia) using fluorescent and standard microscopy to identify the locations of interspecific reproductive isolating barriers. Corymbia torelliana was also crossed with 16 taxa, representing six of the seven Corymbia sections, both Corymbia subgenera and one species each from the related genera, Angophora and Eucalyptus. All crosses were assessed for capsule and seed yields.

Key Results

Interspecific C. torelliana hybridization was controlled by pre-zygotic reproductive isolating barriers inhibiting pollen adhesion to the stigma, pollen germination, pollen tube growth in the style and pollen tube penetration of the micropyle. Corymbia torelliana (subgenus Blakella, sect. Torellianae) was successfully hybridized with Corymbia species from subgenus Blakella, particularly C. citriodora subsp. citriodora, C. citriodora subsp. variegata, C. henryi (sect. Maculatae) and C. tessellaris (sect. Abbreviatae), and subgenus Corymbia, particularly C. clarksoniana and C. erythrophloia (sect. Septentrionales). Attempted intergeneric hybrids between C. torelliana and either Angophora floribunda or Eucalyptus pellita were unsuccessful.

Conclusions

Corymbia hybrids were formed between species from different sections and subgenera, but not with species from the related genera Angophora or Eucalyptus. Reproductive isolation between the interspecific Corymbia hybrid crosses was controlled by early- and late-acting pre-zygotic isolating barriers, with reproductive success generally decreasing with increasing taxonomic distance between parent species. These findings support the monophyly of Corymbia and the close relationships of infrageneric clades. The hybridizing propensity of Corymbia species provides opportunities for breeding but suggests risks of environmental gene flow.     

Leaf flavonoid chemistry and taxonomy of Coreopsis sect. Coreopsis

Seventeen flavonoid glycosides were isolated from the leaves of the nine species of Coreopsis sect. Coreopsis. The compounds include two flavones, four flavonols, a 6-hydroxyflavone, a 6-hydroxyflavonol, two 6-methoxyflavones, a 6-methoxyflavonol and three chalcone-aurone pairs. Chemistry distinguishes all species except C. auriculata and C. intermedia. The mean flavonoid similarity for pair-wise comparison of the four species of annuals (C. basalis, C. nuecensis, C. nuecensoides and C. wrightii) is 0.63 whereas it is 0.66 for the five perennial species (C. auriculata, C. grandiflora, C. intermedia, C. lanceolata, C. pubescens) and 0.54 for comparison of annuals with perennials. This indicates that on the average there is slightly higher flavonoid similarity among species of annuals and among species of perennials than there is between annuals and perennials. Lack of the chalcone-aurone pair coreopsin-sulfurein in perennials is the only consistent chemical difference between annuals and perennials. Leaf flavonoid chemistry supports the hypothesis that C. nuecensis and C. nuecensoides are closely related species but flavonoid data are not concordant with the hypothesis of close relationship between C. basalis and C. wrightii, intermedia, Coreopsis pubescens and C. lanceolata are considered closely related and flavonoid chemistry is concordant with this hypothesis. Coreopsis lanceolata is also viewed as closely allied to C. auriculata, but flavonoids suggest the latter species is more similar to C. intermedia. Coreopsis grandiflora is the only perennial species lacking flavonols and has the fewest number of compounds of any species in the section; chemical data support the view that C. grandiflora is quite distinct from other perennial species of sect. Coreopsis.     

Genetic diversity, origin, and geographic distribution tendencies of biotypes of polyploid spined loaches (Cypriniformes, Cobitidae, Cobitis) in Ukraine

Sixteen of 28 possible biotypes of C. elongatoides-taenia s.lato (C. taenia, C. tanaitica, C. species-1) have been found in Ukrainian waters. Presumably, the biotypes of C. aff. melanoleuca-tanaitica has also been identified. The share of polyploids among Cobitis on average was 65%, where males made up less than 1% with the same ratio among tri- and tetra-polyploids. The absence of amphydiploidy and presence of individuals with recombinant genotype and abnormal electrophoretic specters, whose frequency sometimes is considerable, are attracting increasing interest. There are two centers of polyploid biotypes creation: southern (the Lower Danube) and northern (the Upper Danube, Oder, and Rhine). The C. taenia and C. species-1 type, which is typical for the northwestern basins of Ukraine, participates in the formation of polyploids in the Rhine alongside with C. elongatoides and C. tanaitica. It is determined that biotypes (C. 2(3) elongatoidestanaitica, C. elongatoides-2(3) tanaitica) in comparison with biotypes whose genome includes the chromosome set of C. taenia are limited in distribution towards the East. This is due to the hybridization of females from these biotypes with males of C. taenia afflicted by genetic instability and reduced viability of posterity.     

Does heterospecific seminal fluid reduce fecundity in interspecific copulation between seed beetles?

Reproductive interference through mating between related species can cause fitness reduction and affect population dynamics of the interacting species. In experimental matings between two seed beetles, Callosobruchus chinensis and Callosobruchus maculatus, C. maculatus females, but not C. chinensis females, suffer from significant loss of fecundity when conspecific mating is followed by heterospecific mating. We hypothesized that male traits associated with sexual conflict, which are often harmful to females, pleiotropically affect fitness of heterospecific females through interspecific mating. We examined the effect of ejaculate of C. chinensis males on C. maculatus females as the cause of the fecundity loss in C. maculatus females due to interspecific copulation. We found that frequent interspecific copulation occurred between C. maculatus females and C. chinensis males, but not between C. chinensis females and C. maculatus males, resulting in frequent interspecific ejaculate transfer from C. chinensis males to C. maculatus females. However, injection of the extract from C. chinensis male reproductive organs into C. maculatus females did not significantly affect C. maculatus fecundity compared with saline injection, indicating that the effect of the heterospecific ejaculate transfer on fecundity is negligible. We suggest that other harmful male traits such as genital spines of C. chinensis males are mainly responsible for the fecundity reduction in C. maculatus females that have experienced interspecific mating.     

Histatin 5 Resistance of Candida glabrata Can Be Reversed by Insertion of Candida albicans Polyamine Transporter-Encoding Genes DUR3 and DUR31

Candida albicans and Candida glabrata are predominant fungi associated with oral candidiasis. Histatin 5 (Hst 5) is a small cationic human salivary peptide with high fungicidal activity against C. albicans, however many strains of C. glabrata are resistant. Since Hst 5 requires fungal binding to cell wall components prior to intracellular translocation, reduced Hst 5 binding to C. glabrata may be the reason for its insensitivity. C. glabrata has higher surface levels of β-1,3-glucans as compared with C. albicans; however these differences did not account for reduced Hst 5 uptake and killing in C. glabrata. Similarly, the biofilm matrix of C. glabrata contained significantly higher levels of β-1,3-glucans compared with C. albicans, but it did not reduce the percentage of Hst 5 positive fungal cells in the biofilm. Hst 5 enters C. albicans cell through polyamine transporters Dur3p and Dur31p that are uncharacterized in C. glabrata. C. glabrata strains expressing CaDur3 and CaDur31 had two-fold higher killing and uptake of Hst 5. Thus, neither C. glabrata cell surface or biofilm matrix β-1,3-glucan levels affected Hst 5 toxicity; rather the crucial rate limiting step is reduced uptake that can be overcome by expression of C. albicans Dur proteins in C. glabrata.     

Variation studies on seed storage proteins and phylogenetics of the genus Cucumis

Seed storage proteins were analyzed for variation in polypeptide patterns and proportion of four protein fractions in Cucumis melo and for interrelationships of different taxa of the genus Cucumis. SDS-PAGE of total protein extracts of 11 lines of C. melo representing different geographic regions showed considerable variation in their polypeptides in the range of molecular weights 50–55, 35–39, 20–26 and 16–19 kDa. As compared to one subunit pair reported earlier, two-dimensional gel electrophoresis revealed subunit pairs of molecular weights 53, 52, 50, 42, 39 and 23 kDa, each consisting of a large and a small subunit like those of legumin-like 11S proteins of leguminous seeds. The salt-soluble globulins represented the major protein fraction (44.8–48.1%), followed by glutelins and albumins with prolamins being the lowest in the seed. Based on the protein profiles of seed protein extracts of 28 Cucumis taxa, similarity matrix indices and UPGMA dendrogram showed that eight taxa of C. melo were clustered in two subclusters of four taxa each. Five taxa of C. sativus along with C. zeyheri belonged to another cluster, sister to the third cluster of remaining taxa. In this third cluster, C. anguria var. longaculeatus showed 100% similarity with C. myriocarpus and C. myriocarpus ssp. leptodermis as compared to only 64% similarity with C. anguria; C. myriocarpus ssp. myriocarpus occurred along with C. prophetarum in the same group rather than with C. myriocarpus. The accession PI-299570 representing an undescribed taxon of Cucumis exhibited 100% similarity with C. pustulatus, and C. sagittatus was the most distant from all Cucumis taxa.     

Cytogenetic and genetic data support Crossodactylus aeneus Müller, 1924 as a new junior synonym of C. gaudichaudii Duméril and Bibron, 1841 (Amphibia, Anura)

The nominal anuran species Crossodactylus gaudichaudii Duméril and Bibron, 1841 and Crossodactylus aeneus Müller, 1924 are indistinguishable based on adult and larval morphology, being subject of taxonomic doubts. Here, we describe the karyotypes of C. gaudichaudii and C. aeneus, using classical and molecular cytogenetic markers. In addition, we used sequences of the H1 mitochondrial DNA to infer their phylogenetic relationships by Maximum Likelihood (ML) and Maximum Parsimony (MP) approaches and species delimitation test (by bPTP approach). The karyotypic data do not differentiate C. gaudichaudii and C. aeneus in any of the chromosome markers assessed. In both phylogenetic analyses, C. gaudichaudii and C. aeneus were recovered into a strongly supported clade. The species delimitation analysis recovered the specimens assigned to C. gaudichaudii and C. aeneus as a single taxonomic unit. Taken the cytogenetic and genetic results together with previous studies of internal and external morphology of tadpoles and biacoustic pattern, C. gaudichaudii and C. aeneus could not be differentiated, which supports the hypothesis that they correspond to the same taxonomic unit, with C. aeneus being a junior synonym of C. gaudichaudii.