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The objective of this review was to update the current knowledge on major malaria vectors in Sri Lanka and their bio-ecology and insecticide resistance status. Relevant data were collected through a comprehensive literature search performed using databases such as PubMed, NIH, Google Scholar and Web of Science. Sri Lanka had been endemic to malaria for centuries. However, due to a coordinated public health effort last indigenous malaria case was reported in 2012 and the island nation was declared free of malaria in 2016. Although 25 anopheline mosquitoes have been reported so far on the island, only Anopheles culicifacies and An. subpictus have been established as primary and secondary vectors of malaria respectively. Both vector species exist as a species complex, and the sibling species of each complex differ in their bio-ecology and susceptibility to malaria parasites and insecticides. The article provides a comprehensive and updated account of the bio-ecology and insecticide resistance of malaria vectors and highlights the challenges ahead of retaining a malaria-free status.

Background: The biological characteristics of mosquito vectors vary, impacting their response to control measures. Thus, having up-to-date information on vector bionomics is essential to maintain the effectiveness of existing control strategies and tools, particularly as India aims for malaria elimination by 2030., Objective: This study aims to assess the proportions of vector species resting indoors and outdoors, determine their preference for host biting/feeding, identify transmission sites, and evaluate the susceptibility of vectors to insecticides used in public health programs., Methods: Mosquito collections were conducted in 13 districts across 8 Indian states from 2017 to 2020 using various methods to estimate their densities. Following morphological identification in the field, sibling species of Anopheles mosquitoes were identified molecularly using polymerase chain reaction (PCR)-specific alleles. Plasmodium falciparum and Plasmodium vivax infections in the vectors were detected using enzyme-linked immunosorbent assay (ELISA) and PCR assays. In addition, we assessed the insecticide susceptibility status of primary malaria vectors following the World Health Organization (WHO) protocol., Results: Anopheles culicifacies, a primary malaria vector, was collected (with a man-hour density ranging from 3.1 to 15.9) from all states of India except those in the northeastern region. Anopheles fluviatilis, another primary vector, was collected from the states of Madhya Pradesh, Maharashtra, Karnataka, and Odisha. In Haryana and Karnataka, An. culicifacies sibling species A predominated, whereas species C and E were predominant in Madhya Pradesh and Maharashtra. An. culicifacies displayed mainly endophilic behavior across all states, except in Madhya Pradesh, where the proportion of semigravid and gravid mosquitoes was nearly half of that of unfed mosquitoes. The human blood index of An. culicifacies ranged from 0.001 to 0.220 across all study sites. The sporozoite rate of An. culicifacies ranged from 0.06 to 4.24, except in Madhya Pradesh, where none of the vector mosquitoes were found to be infected with the Plasmodium parasite. In the study area, An. culicifacies exhibited resistance to DDT (dichlorodiphenyltrichloroethane; with <39% mortality). Moreover, it showed resistance to malathion (with mortality rates ranging from 49% to 78%) in all districts except Angul in Odisha and Palwal in Haryana. In addition, resistance to deltamethrin was observed in districts of Maharashtra, Gujarat, Haryana, and Karnataka., Conclusions: Our study offers vital insights into the prevalence, resting behavior, and sibling species composition of malaria vectors in India. It is evident from our findings that resistance development in An. culicifacies, the primary vector, to synthetic pyrethroids is on the rise in the country. Furthermore, the results of our study suggest a potential change in the resting behavior of An. culicifacies in Madhya Pradesh, although further studies are required to confirm this shift definitively. These findings are essential for the development of effective vector control strategies in India, aligning with the goal of malaria elimination by 2030., (©Manju Rahi, AK Mishra, Gyan Chand, RK Baharia, RK Hazara, SP Singh, Siraj Khan, U Sreehari, Divya Kamaraju, Gaurav Kumar, Sanjeev Kumar Gupta, Amit Sharma, K Raghavendra, K Gunasekaran, Om P Singh, Sarala K Subbarao. Originally published in JMIR Public Health and Surveillance (https://publichealth.jmir.org), 17.06.2024.)

The black-throated trogon, Trogon rufus , is a widespread, polytypic species-complex with a convoluted taxonomic history. Here, we integrated morphological, vocal and genetic datasets, including spectral data and digital quantification of barred plumage, to assess and redefine its species limits according to the foremost species concepts. We suggest the recognition of four named and one new species. Trogon tenellus and T. cupreicauda are divergent across Central and South America without geographic overlap or intermediates. Trogon chrysochloros in the Atlantic Forests of Brazil is phenotypically, genetically and ecologically distinct. In Amazonia, Trogon rufus consists of three phenotypically distinct subspecies intergrading with each other in a ring-like formation around central Amazonian rivers. Trogon rufus rufus in the Guiana Shield, Trogon rufus amazonicus in south-eastern Amazonia and Trogon rufus sulphureus in western Amazonia, with contact across the Lower Amazon and Madeira rivers, likely due to secondary contact between incompletely diverged lineages. The unique combination of song, morphology and mtDNA features of an unnamed, isolated population in the Atlantic Forest of north-eastern Brazil resulted in its description as a new species, known only from the type locality and considered here as Critically Endangered, requiring urgent conservation actions. [ABSTRACT FROM AUTHOR]

It is shown that under the old concept of Platyja cyanocraspis Hampson, 1922 at least six species are intermingled, including two sympatric sibling ones from New Guinea. Four such species are new to science and are herein described, namely Platyja subtracta sp. n. (New Guinea), P. togutila sp. n. (Halmahera), P. vityaz sp. n. (Bismarck Archipelago) and P. yaleyambae sp. n. (Louisiades), while a fifth one from Buru and Seram is upgraded from subspecific status as P. lecerfi A.E. Prout, 1922 stat. n. A singleton from New Britain (Bismarck Archipelago) closely albeit not entirely matching P. vityaz is also discussed. [ABSTRACT FROM AUTHOR]

Species delimitation is particularly challenging in Mediterranean nudibranchs because morphological distinction between closely related species is subtle and several cryptic species have been discovered. In this study, we apply an integrative approach to investigate taxonomic distinction of Flabellina cavolini and F. gaditana and assess their genetic and morphologic distinction. We combined a comprehensive phylogeographic survey from across their distribution range and compared genetic variation at mitochondrial and nuclear markers with the variation in colour pattern of cerata that is used to discriminate these two species. Phylogenetic, phylogeographic and species delimitation analyses consistently identify two evolutionary and taxonomic units that do not match with the morphospecies, demonstrating that Flabellina cavolini and F. gaditana are indeed two cryptic, closely related, species and that colour pattern of cerata is not a valid diagnostic character. This study emphasizes how the comparison of range‐wide patterns of phylogenetic and morphological variation, with the inclusion of samples from the type localities, is essential for species delimitation of Mediterranean nudibranchs. Moreover, this study demonstrates the occurrence of Flabellina gaditana in the Mediterranean Sea, allowing to extend the known distribution range of the species, that was previously restricted to the eastern Atlantic Ocean, as well as increasing our inventory of Mediterranean nudibranch diversity. [ABSTRACT FROM AUTHOR]

The flour beetle Tribolium freemani is a sibling species of the model organism and important pest Tribolium castaneum. The two species are so closely related that they can produce hybrid progeny, but the genetic basis of their differences has not been revealed. In this work, we sequenced the T. freemani genome by applying PacBio HiFi technology. Using the well-assembled T. castaneum genome as a reference, we assembled 262 Mb of the T. freemani genomic sequence and anchored it in 10 linkage groups corresponding to nine autosomes and sex chromosome X. The assembly showed 99.8% completeness of conserved insect genes, indicating a high-quality reference genome. Comparison with the T. castaneum assembly revealed that the main differences in genomic sequence between the two sibling species come from repetitive DNA, including interspersed and tandem repeats. In this work, we also provided the complete assembled mitochondrial genome of T. freemani. Although the genome assembly needs to be ameliorated in tandemly repeated regions, the first version of the T. freemani reference genome and the complete mitogenome presented here represent useful resources for comparative evolutionary studies of related species and for further basic and applied research on different biological aspects of economically important pests.

Morphologically similar species, that is cryptic species, may be similar or quasi-similar owing to the deceleration of morphological evolution and stasis. While the factors underlying the deceleration of morphological evolution or stasis in cryptic species remain unknown, decades of research in the field of paleontology on punctuated equilibrium have originated clear hypotheses. Species are expected to remain morphologically identical in scenarios of shared genetic variation, such as hybridization and incomplete lineage sorting, or in scenarios where bottlenecks reduce genetic variation and constrain the evolution of morphology. Here, focusing on three morphologically similar Stygocapitella species, we employ a whole-genome amplification method (WGA) coupled with double-digestion restriction-site associated DNA sequencing (ddRAD) to reconstruct the evolutionary history of the species complex. We explore population structure, use population-level statistics to determine the degree of connectivity between populations and species, and determine the most likely demographic scenarios which generally reject for recent hybridization. We find that the combination of WGA and ddRAD allowed us to obtain genomic-level data from microscopic eukaryotes (-1 millimetre) opening up opportunities for those working with population genomics and phylogenomics in such taxa. The three species share genetic variance, likely from incomplete lineage sorting and ancient admixture. We speculate that the degree of shared variation might underlie morphological similarity in the Atlantic species complex. [ABSTRACT FROM AUTHOR]

Two Norwegian uristid amphipods, obligate associates of sea anemones, have for a long time been confused sub nomine Onisimus normani Sars, 1890. In reality this species only occurs in south Norway, while the north-Norwegian material belongs to O. turgidus (Sars, 1879), described from the Barents Sea and for a long time forgotten. This paper fully illustrates both species, gives a key, and provides data on their distribution and ecology.

For a wide range of insect species, the microbiota has potential roles in determining host developmental programme, immunity and reproductive biology. The tea geometrid moths Ectropis obliqua and E. grisescens are two closely related species that mainly feed on tea leaves. Although they can mate, infertile hybrids are produced. Therefore, these species provide a pair of model species for studying the molecular mechanisms of microbiotal involvement in host reproductive biology. In this study, we first identified and compared the compositions of microbiota between these sibling species, revealing higher microbiotal diversity for E. grisescens. The microbiota of E. obliqua mainly comprised the phyla Firmicutes, Proteobacteria and Cyanobacteria, whereas that of E. grisescens was dominated by Proteobacteria, Actinobacteria and Firmicutes. At the genus level, the dominant microbiota of E. grisescens included Wolbachia, Enterobacter and Pseudomonas and that of E. obliqua included Melissococcus, Staphylococcus and Enterobacter. Furthermore, we verified the rate of Wolbachia to infect 80 samples from eight different geographical populations, and the results supported that only E. grisescens harboured Wolbachia. Taken together, our findings indicate significantly different microbiotal compositions for E. obliqua and E. grisescens, with Wolbachia possibly being a curial factor influencing the reproductive isolation of these species. This study provides new insight into the mechanisms by which endosymbiotic bacteria, particularly Wolbachia, interact with sibling species. [ABSTRACT FROM AUTHOR]

We tested intraspecific as well as interspecific sexual isolation in two sibling species of Drosophila: D. ananassae and D. pallidosa by the multiple-choice technique. The results indicate significant deviations from random mating in both intraspecific as well as interspecific crosses, which demonstrates the existence of strong sexual isolation. Differences in mating propensity in different strains of both the species may be hypothesized as the primary cause behind intraspecific sexual isolation. However, this result was not expected because D. pallidosa is an endemic species that exhibits a greater degree of intraspecific isolation as compared to its cosmopolitan sibling species D. ananassae. Further, both the species show a considerable degree of sexual isolation. We discuss the evolutionary status of both the sibling species in light of these results. [ABSTRACT FROM AUTHOR]

Forficula auricularia (the European earwig) is possibly a complex of cryptic species. To test this hypothesis, we performed: (1) a phylogeographic study based on fragments of the mitochondrial COI and the nuclear ITS2 markers on a wide geographic sampling, (2) morphometric analyses of lineages present in Spain and (3) niche overlap analyses. We recovered five reciprocally monophyletic ancient phylogroups with unique historical patterns of distribution, climatic niches and diversification. External morphology was conserved and not correlated with speciation events, except in one case. Phylogenetic placement of the morphologically distinct taxon renders F. auricularia paraphyletic. Based on the congruence of the phylogenetic units defined by mtDNA and nuclear sequence data, we conclude that phylogroups have their own historical and future evolutionary trajectory and represent independent taxonomic units. Forficula auricularia is a complex of at least four species: the morphologically diagnosable Forficula aeolica González-Miguéns & García-París sp. nov. and the cryptic taxa: Forficula mediterranea González-Miguéns & García-París sp. nov. Forficula dentata Fabricius, 1775 , stat. nov. and Forficula auricularia Linnaeus, 1758 s.s. We also provide new synonymy for F. dentata. [ABSTRACT FROM AUTHOR]

The subspecies of the biological species concept with incomplete reproductive isolation versus the incipient sibling species of the phylogenetic species concept with permeable reproductive barrier are still applied side by side in the everyday practice of taxonomy. Both terms refer to the same organisms diverged mostly in allopatry with various stages of repro-ductive isolation. Question remained: how human ranks these entities organised by nature? The reliable ranking of living hierarchies is retarded and even obscured by the suppressed state of taxonomy. Disappointing scenario: the science of biodiversity is stuck in century old macromorphologies without innovation of fine phenomics and without exploring its high-tech and high-throughput potential. The empirical science of taxonomy is “modernised” by the neutral DNA marker industry diverting the epistemological focus from empirical to virtual. Virtuality of noumenon is used to camouflage the phenomenon of the adverse environmental processes, the wasteful byproducts of the profit oriented liberalized economy. The sensual reality of species and the accelerated species extinction is effectively masked by the virtual sciences of the abstract: numbers, data, statistics, algorithms, equations, models and ideas. To understand the birth of a young incipient species we have briefly reviewed the postmodern development of the unified phylogenetic species concept. (1) The reality of species and higher phylogenetic taxa. (2) The biological and phylogenetic species. (3) How to delineate phylogenetic species? (4) The infinite versus finite division of phylogenetic species. (5) The construct of the unified species concept. (6) Taking subspecies and race out of science. Without recognition of incipient siblings of the phylogenetic species the biodiversity remains under-estimated and the pharisaic anti-science ranking of humans remains with us. The discovery of speciation trait that is the sexual adaptive structures in reproductive barrier building, which are detectable by fine phenomics, gives perspective to find the finite division, the dynamic initial split in the continuous process of diversification. The speciation traits produced by integrative organisation, as opposed to competitive selection, help to unify the operational criteria of the biological species concept that is the speciation by reproductive isolation with the general concept of phylogenetic species that is the causal process of the separately evolving metapopulation lineages. The subspecies and racial ranking is untenable anymore, we suggest taking subspecies and race out of science: the finite division of the initial split detected by speciation traits is the birth of the phylogenetic incipient sibling species. There is no “subspecies”and “races”, as there is no “subindividual” in the biological organisation. In the present caddisfly taxonomy the subspecies remained as a valid status in the Potamophylax cingulatus caddisfly species group. With a clear distinction between the neutral and adaptive traits in the P. cingulatus species group and applying the subtle and stable shape divergences in phallic fine structures we have proposed to change the taxonomic status of subspecies to incipient sibling phylogenetic species rank: Potamophylax alpinus stat. nov., P. depilis stat nov., P. ibericus stat. nov., P. inermis stat. nov., reinstated the species status of P. cingulatus stat. restit. and we have described three new species: P. fesus Oláh, P. portugalicus Oláh et Szczesny, and P. transalpinus Oláh & Coppa, spp. nov.

Understanding the factors that affect hybridization is an important issue in the study of species evolution. In this work, we analyse the genetic structure of two lizard species, Salvator merianae and Salvator rufescens , at a microscale within a climatic niche analysis framework, to reveal the main factors that contribute to the stability of their hybrid zone. We assess the effect of climate in hybridization by quantifying and decomposing the niche overlap of both species. Using a mitochondrial and a nuclear marker, we find that hybridization is frequent and is not restricted to the sympatric region. The gene flow is mainly from S. rufescens to S. merianae , with introgression into the range of S. merianae. Also, S. merianae would have long been present in the area, while S. rufescens appears to be a recent colonizer. The climate contributes to the population structure of S. merianae , but not to that of S. rufescens. The niches occupied by S. rufescens in the hybrid zone and the non-hybrid zone are similar, while the niches of S. merianae are different. Our results do not fit previous models of hybrid zone stability, suggesting the need to develop new models that consider the evolutionary factors that can differentially affect parental species and hybrids. [ABSTRACT FROM AUTHOR]

In a recent integrative taxonomy study, we verified that the previously accepted concept of Lewinskya affinis (≡ Orthotrichum affine) actually comprises a complex of sibling lineages encompassing both known, accepted species (L. affinis s.str., L. praemorsa and L. tortidontia), recovered synonyms (L. fastigiata and L. leptocarpa), and four species yet unpublished. In the present work, we present detailed descriptions of the previously identified species and the new species, L. scissa from the Canary Islands, and the North American L. arida, L. pacifica and L. pseudoaffinis. In addition, we provide a key to the species in the complex, and discuss the morphological distinction of the species according to geographical areas. All included species are illustrated. [ABSTRACT FROM AUTHOR]

Studies assessing the predictability of evolution typically focus on short-term adaptation within populations or the repeatability of change among lineages. A missing consideration in speciation research is to determinewhether natural selection predictably transforms standing genetic variation within populations into differences between species. Here, we test whether and how host-related selection on diapause timing associates with genome-wide differentiation during ecological speciation by comparing ancestral hawthorn and newly formed apple-infesting host races of Rhagoletis pomonella to their sibling species Rhagoletis mendax that attacks blueberries. The associations of 57 857 single nucleotide polymorphisms in a diapause genome-wide-association study (GWAS) on the hawthorn race strongly predicted the direction and magnitude of genomic divergence among the three fly populations at a field site in Fennville, MI, USA. The apple race and R. mendax show parallel changes in the frequencies of putative inversions on three chromosomes associated with the earlier fruiting times of apples and blueberries compared to hawthorns. A diapause GWAS on R. mendax revealed compensatory changes throughout the genome accounting for the earlier eclosion of blueberry, but not apple flies. Thus, a degree of predictability, although not complete, exists in the genomics of diapause across the ecological speciation continuum in Rhagoletis. The generality of this result is placed in the context of other similar systems. This article is part of the theme issue 'Towards the completion of speciation: the evolution of reproductive isolation beyond the first barriers'. [ABSTRACT FROM AUTHOR]

We analysed the COI gene of mitochondrial DNA extracted from larvae of Rabdophaga (Diptera: Cecidomyiidae) that induce rosette galls on Salix in the Holarctic Region. Genetic data show that they belong to species groups of Rabdophaga rosaria and Rabdophaga strobiloides. A maximum likelihood tree indicates that R. rosaria and R. strobiloides populations are divided into clades 1 and 2, respectively. Clade 1 is divided into subclades 1 and 2, and the latter is further divided into subclades 2A and 2B. Subclade 1 consists of populations on several Salix species of section Cinerella in subgenus Vetrix in Georgia and the UK. Subclade 2A contains populations on Salix alba (section Salix , subgenus Salix) in The Netherlands and the UK. Subclade 2B consists of populations on section Helix in Poland, Phylicifoliae in Alaska and Salix species in the Eastern Palaearctic Region. The genetic differences between populations of subclades 1, 2A and 2B range from 1.06 to 3.46%. We propose a possible diversification scenario of R. rosaria into sibling species through the expansion of host plant ranges. Clade 2 consists of R. strobiloides populations on two Alaskan Salix species of the sections Hastatae and Sitchenses in subgenus Vetrix. [ABSTRACT FROM AUTHOR]

Multimammate mice of the genus Mastomys are widespread in sub-Saharan Africa and occur in a wide range of open habitats. Representatives of this genus are the most common African rodents, the main vertebrate agricultural pests and vectors of human pathogens. In Ethiopia, the biogeographically most complex eastern African country, several species have been reported, but their distribution has never been described because of their cryptic morphology. Here we present genetically identified species from 377 Ethiopian Mastomys specimens and analyse their distributional patterns. The genus, represented by four species, inhabits most of the country, with the exception of the highest mountains and dry areas, such as the Afar triangle and the Somali region. For the first time we document M. kollmannspergeri from a single locality in the northernmost part of Ethiopia. Three previously recorded species are more widespread – M. erythroleucus was found at 32 localities, M. natalensis at 13 localities and the Ethiopian endemic species M. awashensis at 18 localities. Phylogenetic analysis of mitochondrial cytochrome b gene sequences indicates that only one of the six phylogroups of M. natalensis and one of the four phylogroups of M. erythroleucus are represented in Ethiopia. Haplotype network analysis indicates two subclades of Ethiopian M. erythroleucus separated by the Ethiopian Rift Valley. Using presence records, we constructed distribution models for the species and analysed the level of overlap. The predicted distribution shows most overlap between M. awashensis and M. natalensis, which is in agreement with empirical data as both species were found in sympatry at four localities. A medium level of overlap was predicted between M. natalensis and M. erythroleucus and both species were found co-existing at two localities. This study not only presents the first detailed distribution of cryptic Mastomys species, but also clearly identifies multimammate mice as model taxa for future evolutionary studies (e.g. the evolution of coexistence or host-parasite interactions) and indicates the regions suitable for such studies. [ABSTRACT FROM AUTHOR]

Astyanax is a species-rich, non-monophyletic genus composed of several supraspecific taxa that are poorly delimited. The Astyanax fasciatus complex is one of these taxa and shows high taxonomic complexity. To elucidate the evolutionary history of the A. fasciatus complex from southern South America, we conducted cytogenetic, molecular and morphological analyses in specimens from the Uruguay River basin. Cytogenetic characters demonstrated two closely related operational taxonomic units: Astyanax sp. 1 (8 m +22 sm +10 st +6 a), Astyanax sp. 2 (8 m +24 sm +10 st +4 a) and natural hybrids (8 m +23 sm +8 st +5 a). 5S ribosomal DNA sites were found in two pairs of m chromosomes and one pair of a chromosomes in Astyanax sp. 1, two pairs of a chromosomes and one pair of m chromosomes in Astyanax sp. 2, and three m chromosomes and three a chromosomes in hybrids. As 51 sites were found in three chromosomes in Astyanax sp. 1 and in five chromosomes in Astyanax sp. 2 and hybrids. Mitochondrial sequence analyses did not separate the two units and hybrids. Morphological analyses revealed differences between Astyanax sp. 2 and hybrids. This secondary contact with gene flow between lineages that diverged long ago might slow or reverse the differentiation/speciation process. These results help us to understand the evolutionary history of this highly complex clade of Astyanax in southern South America. [ABSTRACT FROM AUTHOR]

Background: Anopheles fluviatilis is a species-complex comprising of four cryptic species provisionally designated as species S, T, U and V. Earlier, a 28S-rDNA based allele-specific polymerase chain reaction (ASPCR) assay was developed for the differentiation of the then known three members of the An. fluviatilis complex, i.e., species S, T, and U. This assay was modified in consequence of the discovery of a new cryptic member, species V, in the Fluviatilis Complex to include identification of new species. Methods: In the modified procedure, the ASPCR assay was performed first, followed by restriction digestion of PCR product with an enzyme BamH I, which cleaves specifically PCR amplicon of species V and the resultant PCR–RFLP products can differentiate all the four cryptic members of the complex. Morphologically identified An. fluviatilis samples were subjected to sibling species identification by modified PCR-based assay and standard cytotaxonomy. The result of PCR-based assay was validated through cytotaxonomy as well as DNA sequencing of some representative samples. Results: The modified PCR-based assay differentiates all four sibling species. The result of modified PCR-based assay tested on field samples was in agreement with results of cytotaxonomy as well as DNA sequencing of representative samples. Conclusions: The modified PCR-based assay unambiguously differentiates all four known members of the An. fluviatilis species complex. This assay will be useful in studies related to bionomics of members of the Fluviatilis Complex in their role in malaria transmission. [ABSTRACT FROM AUTHOR]

Using two sibling species, Drosophila jambulina and D. punjabiensis, interspecific crossabilty were examined in natural as well as laboratory conditions. Collection record showed that hybrids are found only in specific hybrid zone having higher temperature variation suggesting an increased fitness in hybrid zone. In laboratory condition, plastic effects on thermo-tolerance traits (heat knockdown and cold recovery) were investigated by growing parental species [D. jambulina and D. punjabiensis at four growth temperatures (17 to 28 °C)] and their hybrids at six growth temperatures (15 to 31 °C). Hybrids showed higher plasticity for melanization (~ 58%) and thermo-tolerance traits (CV = 39.14 for heat resistance and CV = 43.15 for cold resistance) at different growth temperatures. Acclimation effects were also significant in hybrids (F = 47.89 for heat resistance and F = 56.87 for cold resistance) suggesting an adaptive potential. Results of this study imply that hybridization increases genetic variation which in turn leads to adaptation to new environments. The genetic data obtained in this study would be useful not only for testing basic predictions about evolution but also for understanding the evolutionary significance of hybrids. [ABSTRACT FROM AUTHOR]

Morphological stasis or the absence of morphological change is a well‐known phenomenon in the paleontological record, yet it is poorly integrated with neontological evidence. Recent evidence suggests that cryptic species complexes may remain morphologically identical due to morphological stasis. Here, we describe a case of long‐term stasis in the Stygocapitella cryptic species complex (Parergodrilidae, Orbiniida, Annelida). Using phylogenetic methods and morphological data, we find that rates of morphological evolution in Stygocapitella are significantly slower than in closely related taxa (Nerillidae, Orbiniidae). Assessment of quantitative and qualitative morphology revealed the presence of four morphotypes with only subtle differences, whereas molecular data supports 10 reproductively isolated clades. Notably, estimates for the time of Stygocapitella species divergence range from ∼275 million years to ∼18 million years, including one case of two morphologically similar species that have diverged about 140 million years ago. These findings provide evidence for morphological deceleration and long‐term morphological stasis in Stygocapitella, and that speciation is not necessarily accompanied by morphological changes. The deceleration of morphological divergence in Stygocapitella can be potentially linked to niche conservatism and tracking, coupled with the fluctuating dynamics of the interstitial environment, or genetic constraints due to progenetic evolution. Finally, we conclude that failing to integrate speciation without morphological evolution in paleontology may bias estimates of rates of speciation and morphological evolution. [ABSTRACT FROM AUTHOR]

A review of original and published data on invasions of sibling species of Microtus from the group "arvalis" (East European vole, M. rossiaemeridionalis; common vole, M. arvalis; and Altai vole, M. obscurus) is given. Two stages of invasions differing in duration are established. These stages defined the formation of the modern ranges of these species and had an influence on evolutionary processes. The first stage is connected with agricultural development of Eurasia from the early Neolithic to the broad plowing of lands in the second half of the 20th century. Northern expansion of the range following the deforestation for plowing uses and southern expansion as a result of irrigation of arable lands are peculiar to all three sibling species. The second stage is due to expansion of the transport network and growth of traffic in the 20th century. The latter is characteristic of M. rossiaemeridionalis. The natural and historical reasons for the formation a contact zone between M. arvalis and M. obscurus are reconstructed. Original maps of the modern distribution of sibling Microtus species of the "arvalis" group, as well as natural and historical factors that influenced the formation of their ranges, are provided. [ABSTRACT FROM AUTHOR]

Craniometric features (measurements, indexes) in several species of the genus Sylvaemus (S. sylvaticus, S. flavicollis, and S. uralensis from Eastern Europe and some adjacent territories, as well as S. ponticus and S. witherbyi from the Caucasus) have been studied using multivariate statistical analysis. For the first time, craniometric characters of the genus Sylvaemus are considered in relation to the feeding ecology of species. A total of 614 skulls were studied. Based on original material, discriminant keys have been developed using skull measurements. The keys appear to be suitable for the diagnostics of the studied Sylvaemus species, including sibling ones. The largest skulls are shown to be characteristic of S.flavicollis, and the smallest ones, of S. uralensis. Cluster analysis of the absolute measurements of the skull cumulative selection has revealed the greatest isolation of S. flavicollis and the similarity of coexisting but phylogenetically remote S. ponticus and S. witherbyi. Factor analysis showed that, within the genus Sylvaemus, the general sizes are less essential than the characters related to food smashing (the length of the lower jaw and rostrum, the width of the occipital part of the skull), and possibly also to the thickness of the incisors. Comparative analyses of the relative skull measurements (the indexes related to the food preferences of the animals, granivory or folivory) allow us to conclude that the food adaptations to granivory in the structure of the skull and teeth increase in the series S. uralensis–S. sylvaticus–S. witherbyi–S. flavicollis–S. ponticus. The cranial evolution in species within the genus Sylvaemus is suggested to have been directed to increased adaptations in the mechanisms in seed peeling and processing. [ABSTRACT FROM AUTHOR]

Brachionus calyciflorus is arguably the most studied freshwater monogonont rotifer. Although it has been recognised as a cryptic species complex for more than a decade, a formal (re‐)description of the four species known so far (B. calyciflorus, Brachionus dorcas, Brachionus elevatus, and Brachionus fernandoi) has only recently been made. Information on the ecology of these species is very scant and fragmented. The aim of this study was to test for ecological divergence between these four species, specifically their life history strategy and population demography.We conducted a life history experiment using 12–16 genotypes per species. For each species, genotypes were extracted from at least three different natural populations. In addition, we performed population‐level culture experiments with the aim to compare population growth rates and demographic structure of experimental populations among species. Finally, we searched the literature for life history studies with molecular data allowing retrospective species identification.We found pronounced differences in life history traits between B. fernandoi and the other three species. B. fernandoi had higher egg and juvenile development times and a lower egg production rate and mictic ratio. We detected no significant life history differences among B. calyciflorus, B. elevatus, and B. dorcas.Population growth rates of B. fernandoi and B. calyciflorus were higher than those of B. elevatus and B. dorcas. Life history divergence resulted in marked differences in the demographic structure of populations. Populations of B. fernandoi contained larger fractions of pre‐reproductive females and lower fractions of adult females with sexual eggs than populations of B. calyciflorus, B. elevatus, and B. dorcas. Mortality was found to be highest in B. elevatus and lowest in B. calyciflorus populations.Our results show that a reverse taxonomy approach is powerful in revealing sources of variation in ecologically relevant traits of cryptic species, such as life history and demographic structure. Explicit consideration of this variation is crucial for future studies of their dynamics in natural communities. [ABSTRACT FROM AUTHOR]

The water shrew species Neomys fodiens and Neomys milleri are morphologically similar and, based on phenotype, it is difficult to distinguish them. Mandibles of both species were studied by means of classic as well as geometric morphometrics. We compared a priori identified mandibles of water shrews captured in two areas where the species ranges partially or entirely overlap, i.e., in Germany and in the south-eastern part of the Balkan Peninsula. Discrimination of the two species by linear measurements was successful in Germany but not in the Balkans. Discriminant analysis revealed clear differences in the shape of the mandible between the species in both regions. Comparison of allopatric and sympatric populations of both species showed that species are smaller and more similar in shape where they occur in sympatry. Our results suggest that geometric morphometrics can be successfully used for species determination, especially in regions where determination by size is not possible. [ABSTRACT FROM AUTHOR]

Early development of the polychaetes Capitella sp. TF from Tampa, Florida and Capitella sp. TV from Tamiahua Lagoon, Veracruz was studied under laboratory conditions. Our observations indicate that hermaphrodites occur in Capitella sp. TF, but not in Capitella sp. TV. Length and volume of larvae and early juveniles were measured daily. Both species appear to have lecithotrophic development. Sizes and duration of the developmental stages varied widely, as in most known species of Capitella. Characteristic features of Capitella sp. TF include very small hermaphrodites with a distinct dorsal pigmentation pattern and characteristically shaped brood tubes with a centrally thickened region. Embryos forced to abandon the brood did not survive more than 3 days, and handling the brood produced alterations in timing. Ciliated metatrochophores from Capitella sp. TV survived inside the brood without the female for at least 14 days and some larvae hatched and died during settling. Some of these larvae were abnormal, with two rounded protuberances in the two ends. Mortality was high. Fungi and protozoans appeared in the dishes and probably partially induced the production of thicker mucus in the brood tubes, delay in metamorphosis, juvenile mortality, worm length and failure in coupling. Size and duration of the developmental stages were distinctly different from the other putative lecithotrophic species of the complex. These differences and other characteristics of each species discard the possibility that they belong to a previously described species. Our results are applicable to future ecological, morphological and molecular studies of each species, habitat and phylogenetics. [ABSTRACT FROM AUTHOR]

Genus Littorina subgenus Neritrema (Mollusca, Caenogastropoda) includes the “obtusata” group of closely related species (Littorina obtusata and L. fabalis). The anatomy of the adult reproductive system (pallial oviduct) is the only reliable feature used for species identification in females of these species. Reproductive system anatomy and reproduction-associated proteins often diverge between sibling species. Despite being of high evolutionary interest, the molecular basis of this divergence remains poorly understood. We performed proteotranscriptomic comparison of oviducts of L. obtusata and L. fabalis by RNA-seq on Illumina HiSeq 2500 and two-dimensional protein electrophoresis (2D DIGE) with MS/MS identification of the species-specific proteins. The interspecies differences in the oviduct were associated with (1) metabolic proteins reflecting overall physiological differences between L. obtusata and L. fabalis, (2) receptor proteins, and (3) transcripts related to transposable elements (TEs). Various receptors identified may recognize a wide variety of ligands from pathogen-associated molecular patterns to specific carbohydrates on the sperm surface. Therefore, these may participate in immune defense as well as in sperm storage and regulation. Species-specificity of multiple TE sequences (coding for reverse transcriptase and ribonuclease H) may indicate the important role of these genomic elements in the Littorina species divergence, which has not been reported previously.

Abstract Background The Aedes (Stegomyia) albopictus subgroup includes 11 cryptic species of which Ae. albopictus is the most widely distributed. Its global expansion associated with a documented vector competence for several emerging arboviruses raise obvious concerns in the recently colonized regions. While several studies have provided important insights regarding medical importance of Ae. albopicus, the investigations of the other sibling species are scarce. In Asia, indigenous populations within the Ae. albopictus subgroup can be found in sympatry. In the present study, we aimed to describe and compare molecular, morphological and bacterial symbionts composition among sympatric individuals from the Ae. albopictus subgroup inhabiting a Vietnamese protected area. Results Based on morphological structure of the cibarial armarture, we identified a cryptic species in the forest park at Bù Gia Mập in the south-eastern region of Vietnam. Analysis of nuclear (ITS1-5.8S-ITS2) and mitochondrial (cox1, nad5) markers confirmed the divergence between the cryptic species and Ae. albopictus. Analysis of midgut bacterial microbiota revealed a strong similarity among the two species with a notable difference; contrary to Ae. albopictus, the cryptic species did not harbour any Wolbachia infection. Conclusions These results could reflect either a recent invasion of Wolbachia in Ae. albopictus or alternatively a loss of this symbiont in the cryptic species. We argue that neglected species of the Ae. albopictus subgroup are of main importance in order to estimate variation of host-symbionts interactions across evolution.

Correctly numbered plates are provided for the original publication.

The polychaete Capitella sp. A, collected in a NW Mediterranean fish farm (Les Cases d’Alcanar, Tarragona, Spain), was cultured for the first time under experimental conditions with different organically enriched sediments to study the differences in development and growth. The species proved to be dioecious and had lecithotrophic development. Sizes of individuals and duration of the developmental stages varied widely, as in most known species of Capitella. In organically enriched sediments, the juveniles were seen one day after hatching and immature females (i.e. with yellow ovaries) after 52 days. Females may reach maturity (i.e. show white intra-coelomic oocytes) at about 64 days old, and the species had a life span of 167 days. According to its development, Capitella sp. A differs from all known lecithotrophic species of the genus. The results also proved that organically enrichment enhanced growth and survival, whereas lowering food can cause morphological alterations such as reduced size in male genital spines.

A study of phylogenetic relationships in closely related groups of epidemically important species is an important task of parasitology and evolutionary theory. One of these species groups is the maculipennis complex, with its members widespread in Eurasia, North Africa and North America. We studied phylogenetic schemes of the Palaearctic part of this Complex, which are based on crossability of species, polytene chromosomes, isoenzymes, cuticular hydrocarbons and ITS2 of rRNA genes. The lack of high compliance between them prompted us to create a consensus scheme of phylogenetic relationships in the Palaearctic part of this Complex. We constructed phylogenetic schemes on the sequences of a COI gene fragment, the D2 variable region of 28S rDNA and ITS2 of rRNA genes using various algorithms (Neighborjoining, Minimum Evolution, Maximum parsimony and Maximum likelihood). Two consensus schemes of the Palaearctic branch of the maculipennis complex were constructed, validated and discussed. The first scheme was based mostly on molecular genetics data. The second scheme relies on polytene chromosome, geographical spread and ecological data. Phylogenetic relationships of An. beklemishevi are contradictory. This species is similar to other Palaearctic species by molecular markers but it is far from them by fixed chromosome inversions. It was shown that this species is distant from Neoarctic species of the An. quadrimaculatus branch. The proposed schemes show two possible scenarios of the maculipennis complex evolution. Differences between them are explained by facts and associated speculations as well as by assumptions made under different phylogenetic reconstruction methods. A hypothesis about a significant role of the disparity between the astronomical and biological times in the interpretation of evolutionary processes was proposed. To address the phylogeny problem of the Palaearctic branch of the maculipennis complex at a more detailed level, it is required to increase our knowledge about the underlying species.

Abstract Among six dominant mosquito vector species involved in malaria transmission in India, Anopheles minimus is a major species in northeast India and held responsible for focal disease outbreaks characterized by high-rise of Plasmodium falciparum infections and attributable death cases. It has been now genetically characterized that among the three-member species of the Minimus Complex spread in Asia, An. minimus (former species A) is prevalent in India including northeastern states and east-central state of Odisha. It is recorded in all seasons and accounts for perennial transmission evidenced by records of sporozoite infections. This species is highly anthropophilic, and largely endophilic and endophagic, recorded breeding throughout the year in slow flowing seepage water streams. The populations of An. minimus in India are reported to be highly diverse indicating population expansion with obvious implications for judicious application of vector control interventions. Given the rapid ecological changes due to deforestation, population migration and expansion and developmental activities, there is scope for further research on the existence of potential additional sibling species within the An. minimus complex and bionomics studies on a large geographical scale for species sanitation. For control of vector populations, DDT continues to be applied on account of retaining susceptibility status even after decades of residual spraying. Anopheles minimus is a highly adaptive species and requires continuous and sustained efforts for its effective control to check transmission and spread of drug-resistant malaria. Anopheles minimus populations are reportedly diminishing in northeastern India whereas it has staged comeback in east-central State of Odisha after decades of disappearance with its eco-biological characteristics intact. It is the high time to siege the opportunity for strengthening interventions against this species for its population diminution to sub-optimal levels for reducing transmission in achieving malaria elimination by target date of 2030.

A study was undertaken in the villages of Korea and Bastar district (Chhattisgarh) during the years 2012–2015 to investigate the bionomics of malaria vectors and the prevalence of their sibling species complexes. Entomological surveys carried out every month included indoor resting collections, pyrethrum spray catches, light trap catches, and insecticide susceptibility status of Anopheles culicifacies using World Health Organization (WHO) methods. Anopheles culicifacies and Anopheles fluviatilis species were assayed by polymerase chain reaction (PCR) for the detection of malaria parasite, and sibling species were identified using PCR and DNA sequencing. A total of 13,186 samples of Anopheles comprising 15 species from Bastar and 16 from Korea were collected. An. Culicifacies was recorded as the most dominant species and also the only active vector at both sites. This species was found to be resistant to dichlorodiphenyltrichloroethane (DDT) and Malathion, showing signs of emerging resistance against pyrethroids. Among the sibling species of An. culicifacies, the group BCE was found in maximum numbers, while sibling species T of the An. fluviatilis was recorded to be dominant among its complex. The study provides a comprehensive view of the vector bionomics in the highly malarious regions of India that may have importance in developing vector control strategies.

The mosquito Anopheles (Cellia) subpictus sensu lato (s.l.) is a major secondary vector of malaria in Sri Lanka. The sibling species composition in this species complex in Sri Lanka remains debatable. Compensatory base changes (CBCs) in the secondary structures of internal transcribed spacer 2 (ITS2) are reliable sources to predict sexual incompatibility among closely related species. The objective of the present study was to investigate the An. subpictus s.l. populations in Sri Lanka using the CBC analysis. Mosquito DNA was amplified and sequenced for the ITS2 region. The sequences were annotated using ITS2 Database. ITS2 secondary structures were constructed and analyzed for CBCs using various bioinformatics tools. The ITS2 regions consisted of two different lengths, 575 bp and 480 bp. The two CBCs and three hemi CBCs identified in the present study suggest that there may be at least two sexually incompatible sibling species. In conclusion, it is likely that there may be only two reproductively isolated sibling species in the An. subpictus species complex in Sri Lanka. However, due to high divergence of ITS2 in these species, it is reasonable to assume that they may be undergoing a speciation event to separate as a distinct species.

The genus Matuxia Carbayo et al., 2013 currently comprises two species with distribution restricted to southeastern Brazil. In the present study, based on an integrative approach, we examine the genetic diversity within the genus and describe a new species, Matuxia tymbyra Rossi and Leal-Zanchet, sp. nov., representing a southern lineage of the genus. We employed one mitochondrial (cytochrome c oxidase subunit I, COI) and other nuclear (elongation factor 1a, EF-1a) markers to investigate the phylogenetic relationships within the genus. Maximum-parsimony analysis in a segment of the COI gene of 676 nucleotides showed 79 (11.68%) nucleotide positions exhibiting autapomorphic characters that support the occurrence of three independent evolutionary molecular operational taxonomic units in the genus. Similar evaluation for the dataset of the EF-1a gene showed a much smaller number of autapomorphies. Bayesian inference, maximum-likelihood and delimitation approaches, based on evolutionary models, showed that M. tymbyra is the sister species of the type-species of the genus, Matuxia tuxaua. They should be considered sibling species, only distinguishable based on details regarding eye arrangement and prostatic vesicle. The existence of 15 and 23 molecular autapomorphies, as revealed by maximum-parsimony analysis in a segment of the COI gene for M. tymbyra and M. tuxaua, respectively, allowed us to propose a molecular diagnosis for the new species, which is essential in cases of sibling species. The new species seems to be endemic from areas of Araucaria Forest in southern Brazil; the record augments the known distribution of the genus to the south. [ABSTRACT FROM AUTHOR]

The Myotis myotis/M. blythii species complex, spread across the Western Palearctic, is a problematic group for which the taxonomy of the species is not yet satisfactorily resolved. The Iberian Peninsula played a key role in its evolutionary history as a Pleistocene refuge and as the starting point for the eastward expansion of M. myotis in the early Holocene, while M. blythii reached the Iberian Peninsula only during the middle Holocene. The study of Iberian populations and particularly of the Iberian fossil record is of high interest in this regard. However, there are few data available on the biometry of the skulls and teeth of Iberian populations (which differ somewhat in size from those of other regions of Europe and Asia) or tools for the identification of fragmentary cranial remains. Much of the Quaternary Iberian record of large Myotis remains unassigned. Here, we contribute to the task of determining fragmentary cranial remains by providing new cranial and dental biometric data from extant Iberian populations, predictive models for isolated upper molar identification, and a set of indices that allow quantitative evaluation of the differences in anatomical traits (in skull and molars) between the two species. [ABSTRACT FROM AUTHOR]

Aquatic species are facing at higher risk of extinction similar to that of any other living components of diversified ecosystem in present scenario. So that, the conservation of aquatic biodiversity is much more important to know about the accurate information regarding species composition and their biological community interactions. Generally, traditional survey methods depend on physical identification and characterization of species but it has some sorts of challenging chances due to the phenotypic plasticity, sibling species, different stages of life cycle and its invasiveness. To overcome such barriers one of the significant and promising tool likewise environmental DNA (eDNA), which way the collection of genetic materials from bulk environment (i.e. soil, water, sediment etc.) circuitously from organisms has been used to monitor and analyzed the biodiversity status, invasive species along with the species of conservation category. Recently, the real application of eDNA analysis based outcomes uphold the actual emerging know how practices in support of the population and community ecology, conservation biology as well as in the superior field of taxonomical research. Such scientific appraisal will be useful in understanding the brief history of aquatic eDNA and obviously its methodological considerations, gentle sources, collection and analysis process, physical form, its persistence and proper transport in aquatic ecosystem. Moreover, the fruitful drives for summarization the discoveries of eDNA application and method over traditional technique, its recent challenges and examine the current and future frontiers along with the appropriate practices of aquatic eDNA relevancy in aquatic ecosystem. [ABSTRACT FROM AUTHOR]

Chrysoperla comanche (Banks) and its sibling species Chrysoperla rufilabris (Burmeister) are voracious predators of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), an invasive pest that vectors a bacterium responsible for the lethal and incurable citrus greening disease. The Comanche lacewing naturally occurs in Southern California, whereas C. rufilabris is currently one of only two commercially available green lacewing in the United States. These sister species can be separated by larval morphological traits, by differences in courtship songs, and possibly by three nuclear genes wingless, PepCK, ATPase , yet they are not distinguishable based on the mitochondrial barcode gene (COI). Releasing in a new area a biological control agent capable of hybridizing with a resident species may pose risks that range from local displacement to irreversible loss of genetic identity. Therefore, we performed no-choice laboratory crosses to assess pre- and postzygotic isolation. We show that fertile and viable hybrid progeny could be readily obtained in interspecific crosses and backcrosses and, although there is a trend toward lower hybrid fitness, postzygotic isolation is overall weak and might not prevent loss of genetic identity under natural conditions. It remains to be determined if differences in courtship songs will prevent hybridization in the wild, as shown for other green lacewings. We also report a low prevalence of Rickettsia infection in both species. [ABSTRACT FROM AUTHOR]

Chrysoperla duellii sp. nov. is discovered, associated with coniferous vegetation, in the Kyrgyz Republic in central Asia, co-occurring with two other cryptic species of the Chrysoperla carnea-group. It is distinguished by its courtship duetting song, which exhibits a uniquely complex multi-volleyed temporal and tonal structure but is nevertheless noticeably convergent on the species-specific song of another conifer-associated member of the carnea-group, C. downesi (Smith), from North America. The song of C. duellii also shows some superficial similarity to those of European/north African C. mediterranea (Hölzel) and southern Californian C. calocedrii Henry et al., both of which, like C. duellii, prefer conifers and exhibit dark-green adult body pigmentation. The new species is herein formally described as Chrysoperla duellii Henry, Taylor and Johnson and is shown to be diagnosable by song and morphology from all other known members of the Chrysoperla carnea-group. Results of restriction site-associated DNA sequencing (RADseq) of C. duellii and five other carnea-group species strongly reject the hypothesis that similar songs and dark-green body colour among the four conifer-associated species are due to common ancestry, instead supporting independent, parallel acquisition of such features. [ABSTRACT FROM AUTHOR]

Population genetics is a convenient tool to study the population biology of non‐model and hard to sample species. This is particularly true for parasites and vectors. Heterozygote deficits and/or linkage disequilibrium often occur in such studies and detecting the origin of those (Wahlund effect, reproductive system or amplification problems) is uneasy. We used new tools (correlation between the number of times a locus is found in significant linkage disequilibrium and its genetic diversity, correlations between Wright's FIS and FST, FIS and number of missing data, FIT and allele size and standard errors comparisons) for the first time on a real data set of tsetse flies, a vector of dangerous diseases to humans and domestic animals in sub‐Saharan Africa. With these new tools, and cleaning data from null allele, temporal heterogeneity and short allele dominance effects, we unveiled the coexistence of two highly divergent cryptic clades in the same sites. These results are in line with other studies suggesting that the biodiversity of many taxa still largely remain undescribed, in particular pathogenic agents and their vectors. Our results also advocate that including individuals from different cohorts tends to bias subdivision measures and that keeping loci with short allele dominance and/or too frequent missing data seriously jeopardize parameter's estimations. Finally, separated analyses of the two clades suggest very small tsetse densities and relatively large dispersal. [ABSTRACT FROM AUTHOR]

Anisakid nematodes have a worldwide distribution and are associated with fishes, birds, and marine mammals from freshwater, brackish, and marine systems. The aims of this work are to report for the first time Contracaecum mirounga parasitizing the Magellanic penguin Spheniscus magellanicus, to report another Contracaecum species in the same host species, and to discuss the validity of Contracaeceum spheniscus. Several dead chicks, juveniles, and adults of S. magellanicus were collected along the Argentinean coast from 2002 to 2009. Nematodes were removed from digestive tracts and studied using both light and scanning electron microscopy. Nematode prevalences were 2.38% for C. mirounga and 12.5% for Contracaecum sp. Contracaecum mirounga was found in 1 penguin from Península Valdés, Chubut. This species is known as a specific parasite of marine mammals such as Pinnipedia, thereby suggesting that this nematode is not as specific as believed. Another species of Contracaecum sp. was found parasitizing 1 penguin from the Río de la Plata coast. It possessed an unusual interlabial morphology and arrangement of male caudal papillae. Despite the low prevalence, the distinct morphological features are convincing and support the presence of a new Contracaecum species. However, a formal description is not presented because sufficient male specimens are lacking. Finally, C. spheniscus is considered a junior synonym of Contracaecum pelagicum. Future molecular studies might be helpful to determine the real diversity of Contracaecum species parasitizing S. magellanicus considering the number of sibling species recognized among the anisakids. [ABSTRACT FROM AUTHOR]

We investigated the main environmental variables determining the occurrence of the Eurasian Treecreeper Certhia familiaris and Short-toed Treecreeper C. brachydactyla within the mountain range of the Pyrenees. Presence/absence data were obtained, using the response to the playback of each species' typical song; they were collected during the springs of 2004-2012 over 487 sites, within 33 areas of mature forest distributed over the mountain range. The analysis, based upon generalised linear models, showed that altitude, forest type, location within the mountain range and the presence of the sibling species influenced the occurrence of both treecreepers. Probability of occurrence of the Eurasian Treecreeper increased with altitude except in mountain pine forest, whereas at a given altitude it was lower in Scots pines than in other forest types. In the Short-toed Treecreeper, whose probability of occurrence was enhanced in oak forest, the altitude had a negative influence except in mountain pine forest. In the Eurasian Treecreeper, all other things being equal, probability of occurrence was higher in the forested areas experiencing an oceanic climate than those experiencing a Mediterranean-continental climate. The opposite was found in the Short-toed Treecreeper, whose probability of occurrence on the Pyrenees' southern slope increased from the western to the eastern end of the mountain range. Finally, the presence of the sibling species decreased the probability of occurrence of both treecreepers at low and medium altitude, suggesting a restricted form of interspecific competition. [ABSTRACT FROM AUTHOR]

A morphological, molecular, and behavioural characterization of Rhionaeschna galapagoensis is presented, based on a series of specimens and observations from San Cristóbal Island, Galápagos, including both adults and larvae. Several of the characters proposed earlier to distinguish between the adults of this species and its closest relative, R. elsia, are found to be variable, but the presence of a black band over the fronto–clypeal suture is confirmed as a good diagnostic character. The ultimate instar larvae of R. galapagoensis is described for the first time, and diagnosed from its closest relatives by a combination of characters, including the acute angle between the prothoracic apophyses, absence of lateral spines on abdominal segment 6, and length of cerci relative to paraprocts. Molecular analysis confirmed that R. galapagoensis and R. elsia are sister species, and showed that their genetic distance is the closest among the analyzed species, which is to be expected given the young age of the Galápagos Islands. The larvae of R. galapagoensis were very common and widespread in the mountain streams and a pond in the southwest of San Cristóbal. Swarms of tens of individuals formed at sunrise in the coastal vegetation, together with adults of Tramea cf. cophysa, feeding on small flying insects. Males showed patrolling behaviour on small sections of the streams and at a pond. Only one copulation was observed, lasting 10 minutes. Females oviposited alone on floating vegetation in running and standing waters. Our observations corroborate that R. galapagoensis and R. elsia are two parapatric species, morphologically and genetically close. In San Cristóbal, R. galapagoensis had large populations, apparently not threatened.

Abstract Background Anopheles gambiae s.s. and An. coluzzii are two of the most important malaria vector species in sub-Saharan Africa. These recently-diverged sibling species do not exhibit intrinsic post-zygotic barriers to reproduction and are thought to be separated by strong assortative mating combined with selection against hybrids. At present, little is known about the ecological conditions that determine hybridization and introgression between these cryptic taxa. Methods Swarm segregation and assortative mating were studied in Western Burkina Faso in the villages of Vallée du Kou (VK7) and Soumousso which differed in terms of which sibling species was much rarer than the other, and in Bana where both occurred in similar proportions. Swarms and pairs in copula were collected and genotyped, the proportion of intra and interspecific mating determined, and interspecific sperm transfer checked genetically. Females were collected through larval and adult indoor collections and genotyped or sexed-and-genotyped via a novel multiplex PCR. Results A total of 3,687 males and 220 females were collected and genotyped from 109 swarms. Only 3 swarms were composed of males from both species, and these were from the village of VK7 where An. gambiae s.s. was comparatively rare. Mixed-species pairs captured in copula were only detected in that area and made for 3.62 % and 100 % of mating pairs involving An. coluzzii and An. gambiae s.s. individuals, respectively. The high An. gambiae s.s. cross-mating rate was mirrored by high rates of hybridizations estimated from female larvae and adults indoor collections. This contrasted with Soumousso, where despite being much less common than An. gambiae s.s., An. coluzzii males did not form mixed swarms, females were not found in interspecific swarms or copula and hybridization rates were low in both sibling species. Conclusions These data suggest that ecological conditions leading to rare An. gambiae s.s. in populations dominated by An. coluzzii may promote a breakdown of spatial swarm segregation and assortative mating between the two species. The lower overall hybridization rates observed at the larval and adult indoor stages compared to cross-mating rates support the idea that post-mating selection processes acting against hybrids may occur mostly prior to and/or at the early larval stages.

A study of the relationships between 21 southern European, Moroccan and Turkish populations of the Merodon avidus species complex was carried out. Based on a parallel study of type material from several museums, documented diagnostic morphological characters, season of adult activity and geographical distribution, we justify the use of the following names for three closely related taxa in this complex: M. avidus (Rossi, 1790), M. moenium (Wiedemann, 1822), and M. ibericus Vujić nom. n. (new name for M. bicolor Gil Collado, 1930, preoccupied by M. bicolor Walker, 1852). Neotypes of Merodon avidus, M. spinipes (Fabricius, 1794) and M. quadrilineatus Lioy, 1864 are designated. Lectotype of Merodon moenium (Wiedemann in Meigen, 1822) is designated here. A cluster analysis of DNA barcoding sequences clearly separated M. ibericus, but not M. avidus and M. moenium, even though the lack of shared haplotypes, analysis of molecular variance (AMOVA), pairwise Φst values together with allozyme and ecological niche analyses revealed statistically significant percentage of variation among all three species in the Merodon avidus complex. Analysis of 5 diagnostic enzyme loci revealed the presence of genetic differentiation among the M. avidus/moenium complex populations investigated (Fst = 0.654) and species-specific alleles were found at the AAT locus. The presence of two separate related taxa within the M. avidus/moenium complex was further supported by an UPGMA tree based on Nei's (1978) genetic distances. The value of Nei's measure of genetic identity (I = 0.520) between two large (meta) populations of M. avidus and M. moenium suggest that these taxa are sibling species. Populations from Djerdap (Serbia) confirmed the presence of temporal divergence between these species at a locality where they occur sympatrically, while spring and autumn populations from Umag (Croatia) provide an example of morphological plasticity within the species M. avidus. Ecological niche analysis contributed to the species delimitation. Review of the available genetic and ecological data confirmed our hypothesis that the M. avidus species complex, in addition to M. ibericus Vujić nom. n. from the Iberian Peninsula, consists of two sibling species in the rest of Europe and indicated their recent speciation.

Paramecium (Ciliophora) is an ideal model organism to study the biogeography of protists. However, many regions of the world, such as Central America, are still neglected in understanding Paramecium diversity. We combined morphological and molecular approaches to identify paramecia isolated from more than 130 samples collected from different waterbodies in several states of Mexico. We found representatives of six Paramecium morphospecies, including the rare species Paramecium jenningsi, and Paramecium putrinum, which is the first report of this species in tropical regions. We also retrieved five species of the Paramecium aurelia complex, and describe one new member of the complex, Paramecium quindecaurelia n. sp., which appears to be a sister species of Paramecium biaurelia. We discuss criteria currently applied for differentiating between sibling species in Paramecium. Additionally, we detected diverse bacterial symbionts in some of the collected ciliates.