Mitochondrial DNA Barcode Research Articles

Anomalous latitudinal gradients in parasitoid wasp diversity-Hotspots in regions with larger temperature range.

Knowledge of global patterns of genetic diversity is essential for biodiversity conservation as this parameter describes the ability of a species to respond to environmental changes. Ichneumonoids parasitoid wasps are among the few taxa showing an anomalous latitudinal diversity gradient. Using the largest georeferenced molecular dataset for this group, we used a macrogenetics approach to examine latitudinal patterns and predictors of intraspecific genetic diversity. We calculated the mean nucleotide diversity of mitochondrial DNA barcode sequences at three geographic levels: grid cells, latitudinal bands and climatic zones. Nucleotide diversity values were consistently higher at northern temperate latitudes, peaking at 50°. We found a positive but weak relationship between intraspecific diversity and the latitude, between intra- and interspecific diversity, and a positive effect of the temperature range. Examining the spatial relationship between different levels of biodiversity and its drivers is particularly relevant considering climate change and its impact on species distribution. Yet, in insects, it has been challenging to integrate ecological, evolutionary and geographical components when analysing the processes leading to species richness gradients.

Tracing the origin of the alien pest Cydia pomonella in Algeria through a worldwide comparison of the species’ DNA barcodes

Abstract DNA barcodes (i.e., gene sequences used as identifiers for species identification) constitute a very useful tool in invasive pest research. Using them, we studied, for the first time, the origin and population structure of the alien Cydia pomonella in Africa, a major apple pest. We sequenced a fragment of the mitochondrial gene Cytochrome c oxidase subunit I (COI) from C. pomonella caterpillars collected in northern Algeria and compared them with DNA barcodes from the rest of the world available at public repositories. The phylogeny built upon that COI database supports the European origin of the species: haplotypes at the base of the phylogeny were registered in this continent. In Algeria, the results suggest that this pest could have been introduced from Europe, with two unique African mutations probably favoured by C. pomonella multivoltinism and large population sizes. Population genetic analyses at three Algerian localities showed neither structure nor founder effects. However, to confirm the underlying process of the observed population patterns, it is necessary to perform analyses using genes with higher mutation rates. Compared to higher resolution markers, mitochondrial DNA barcodes are still a cost‐effective tool for taxonomic determination of unknown specimens and, sometimes, may also allow a preliminary tracing of its origin. This is crucial to detect and stop the spread of invasive pests (i.e., borders). We encourage further studies including functional genes to assess whether any mutations are promoting C. pomonella adaptation to the Northern Africa environment.

A Microcosm Analysis of Species-Specific Responses of Chironomidae on Heavy Metal Pollution in The Nyanza Gulf of Lake Victoria

The Chironomidae family, known as "non-biting midges" in their adult stage and "bloodworms" in their larval stage, consists of diverse dipteran insects inhabiting various global aquatic environments. Despite extensive global research, data on Chironomidae in the polluted Nyanza Gulf of Lake Victoria, Kenya, is scarce, and molecular identification methods have not been explored. This study aimed to quantify heavy metal concentrations in water, sediment, and insect samples and assess their impact on Chironomid species identified using mitochondrial DNA barcoding of the cytochrome oxidase subunit 1 (COI) gene. Analysis of Variance was used to determine if there were any statistically significant differences in heavy metal concentrations across different sample types or locations along the pollution gradient. Chironomids were collected from Nyanza Gulf, focusing on a pollution gradient. Results showed that concentrations of arsenic (As), lead (Pb), and cadmium (Cd) in insect, water, and sediment samples exceeded standard limits, while mercury (Hg) concentrations were within limits. Significant variations (p ≤ 0.05) in Pb levels were observed in water samples, and heavy metal concentrations in sediment samples varied significantly (p ≤ 0.05), with Pb showing the highest variation (p ≤ 0.0001). Insect samples exhibited significant differences (p ≤ 0.0001) in As and Hg contents. Genetic analysis identified two known species: Chironomus transvaalensis at the heavily polluted Kisumu station and Chironomus pseudothummi at the moderately polluted Kendu Bay and Homa Bay stations. Additionally, a unique Chironomus species was found on Ndere Island, a relatively clean site with restricted human activities. Sequence comparisons indicated proximity to global data but also highlighted the evolutionary significance and uniqueness of the identified species. This study demonstrated the potential use of genetic methods in determining Chironomid species diversity, community structure, and abundance in relation to heavy metal concentration. It suggests that heavy metal pollution may act as a selective pressure, driving the evolution of Chironomid species. The study recommends combining genetic approaches with other pollution sources for a comprehensive understanding of using this species in monitoring pollution

Marine catfishes (Ariidae-Siluriformes) from the Coastal Amazon: mitochondrial DNA barcode for a recent diversification group?

Ariidae species play a significant role as fishing resources in the Amazon region. However, the family's systematic classification is notably challenging, particularly regarding species delimitation within certain genera. This difficulty arises from pronounced morphological similarities among species, posing obstacles to accurate species recognition. Following morphological identification, mitochondrial markers (COI and Cytb) were employed to assess the diversity of Ariidae species in the Amazon. Our sampling efforts yielded 12 species, representing 92% of the coastal Amazon region's diversity. Morphological identification findings were largely corroborated by molecular data, particularly for species within the Sciades and Bagre genera. Nonetheless, despite morphological support, Cathorops agassizii and Cathorops spixii displayed minimal genetic divergence (0.010). Similarly, Notarius quadriscutis and Notarius phrygiatus formed a single clade with no genetic divergence, indicating mitochondrial introgression. For the majority of taxa examined, both COI and Cytb demonstrated efficacy as DNA barcodes, with Cytb exhibiting greater polymorphism and resolution. Consequently, the molecular tools utilized proved highly effective for species discrimination and identification.

Larval Taxonomy of the Caddisfly Cernotina truncona Ross, 1947 (Trichoptera: Polycentropodidae)

The genus Cernotina Ross, 1938 is represented in the southeastern United States by three nominal species: Cernotina calcea Ross, 1938, Cernotina spicata Ross, 1938, and Cernotina truncona Ross, 1947. Of all Cernotina species, only the larva of C. spicata has been described to date. The goal of this paper is to describe, illustrate, and diagnose the larva of C. truncona using ecologically associated specimens. In addition, we used publicly available mitochondrial DNA barcoding data to evaluate the genetic relationships of these species. The larvae of Cernotina truncona can be distinguished from those of C. spicata by differences in setal placement and number on the meso- and metanota, mandibular morphology, head width, and distal setation of the tarsi. The ultrastructure of the anal claw is figured, highlighting the novel finding of small spines on the concave margin of a larva of Cernotina. With this new description, just the second described larva from this genus, only C. calcea remains unknown in the southeastern United States. The information provided herein enables the in-depth study of the ecology and life history of this diminutive caddisfly.

Cryptic Taxa Revealed through Combined Analysis of Chromosomes and DNA Barcodes: The Polyommatus ripartii Species Complex in Armenia and NW Iran.

The detection of cryptic species in complexes that have undergone recent speciation is often difficult, since many standard nuclear markers have not yet accumulated differences between closely related taxa, and differences in mitochondrial markers can be leveled out due to mitochondrial introgressions. In these cases, the use of derived chromosomal characters such as non-ancestral chromosomal numbers and/or unusual karyotype features may be a solution to the species delimitation problem. However, non-ancestral but similar karyotypes may arise secondarily as a result of homoplastic evolution, and their interpretation as homologies may lead to incorrect taxonomic conclusions. In our study, we show that the combined use of mitochondrial DNA barcodes and karyotypes helps to solve this problem and identifies cryptic species in situations where each of these markers does not work individually. Using this approach, we show that the fauna of Armenia and adjacent Iran includes the following cryptic taxa of the Polyommatus ripartii species complex (haploid chromosome number, n in parentheses): P. ripartii paralcestis (n = 90), P. ripartii kalashiani, subsp. nov (n close to 90), P. emmeli, sp. nov. (n = 77-79), P. keleybaricus, sp. nov. (n = 86), P. demavendi belovi (n = 73-75), P. demavendi antonius, subsp. nov. (n = 71-73), P. admetus anatoliensis (n = 79) and P. eriwanensis (n = 29-34). Polyommatus admetus yeranyani is synonymized with P. admetus anatoliensis.

Challenges of molecular barcode-based identification of earthworm specimens for biodiversity assessment

Specimen identification at the species level is a critical challenge for understanding community structure and conserving biodiversity. The use of mitochondrial DNA barcodes, in addition to morphology, has proven to be a useful tool for earthworm identification, but it has also raised difficulties. Thus, approaches to delineating molecular operational taxonomic units (MOTUs) from cytochrome c oxidase subunit 1 (COI) or 16S rDNA sequence data often reveal more mitochondrial lineages than morphospecies, raising the question of whether these MOTUs should be used as taxonomic entities in community structure studies. Here, we used a newly acquired dataset of 576 COI barcodes of earthworms from Metropolitan France that were clustered in 36 MOTUS, corresponding to 21 morphospecies. We also incorporated data gathered from the literature to investigate this question. In order to match our MOTUs with already-described mitochondrial lineages, we downloaded reference sequences from the GenBank and BOLD platforms. In light of the difficulties encountered in recovering these sequences, we recommend that any new mitochondrial lineage described in a study be named consistently with previous works. Next, we analyzed the biological, ecological, and molecular data available in the literature on the different mitochondrial lineages that matched our MOTUs in order to determine if there was a consensus for species delimitation. Although the study specimens mainly belong to the Lumbricidae, which is one of the most studied families of earthworms, the data are often missing to determine if the MOTUs correspond to different species. Pending revision of the taxonomy, MOTUs for which mitochondrial divergence has been confirmed by morphological differences should be considered distinct taxonomic entities in community structure studies. In the absence of morphological differences and pending more data, we propose to distinguish for these analyses, within sexually reproducing morphospecies, the MOTUs for which mitochondrial divergence has been confirmed by multilocus nuclear data, while in the case of reproduction by parthenogenesis, ecological differences between MOTUs are necessary to consider them as different taxonomic entities.

Even one tardigrade is enough. Molecular methods revealed presence of pantropical species <em>Echiniscus lineatus</em> (Heterotardigrada, Echiniscidae) in the fauna of Seychelles

A brief report on the tardigrade species Echiniscus lineatus discovered in a moss cushion collected in Seychelles is given. The species represents a new pantropical element in the tardigrade fauna of Seychelles. With this record the number of echiniscid tardigrade species in Seychelles has reached five. Morphological identification of the species is supported with the analysis of obtained sequences of nuclear and mitochondrial DNA barcode genes (ITS-1, ITS-2 and COI). Analysis of the genetic data using the method of TCS haplotype networks revealed separated position of the Seychellian population of E. lineatus and supported the hypothesis of independent colonisation of Seychelles.

Investigating Calico Crayfish (Faxonius immunis Hagen, 1870) as a possible “sleeper” invasive species in northern Wisconsin, United States

“Sleeper” invaders are non-native populations that experience time-lags post-establishment before subsequent spread or negative impacts, challenging managers to differentiate harmless non-native species from invasive species. In lakes of northern Wisconsin, United States, Rusty Crayfish (Faxonius rusticus Girard, 1852) has dominated as an invasive species for decades, but this species has recently experienced population declines. Following these F. rusticus declines, we rediscovered in 2020 a population of non-native Calico Crayfish (Faxonius immunis Hagen, 1870) that had not been documented since the 1970s. Declining F. rusticus populations may create opportunities for F. immunis spread to other lakes and impacts as a sleeper invader. We conducted additional sampling in summer 2021 that suggests F. immunis remains isolated in only one lake within this watershed. We used mitochondrial DNA barcoding to confirm these crayfish were F. immunis and had not been misidentified as a congener. Next, we investigated whether biotic interactions with F. rusticus may have prevented F. immunis spread over the past several decades. We measured agonistic behaviors using F. immunis and F. rusticus pairs in the laboratory, and then modeled differences in aggression between species while controlling for size and reproductive form. We found that F. rusticus were consistently dominant over F. immunis, suggesting that competition with an established hyper-abundant invasive species may have restricted past spread by F. immunis. Managers and policy makers should consider whether precautionary actions against F. immunis are warranted while the population of this species remains small and localized, especially in the context of F. rusticus declines.

Advancing One Health in Urban Seafood Markets: A Genetic and Social Analysis of Dried Sea Cucumber in Three New York City Chinatowns

This study employs a multidisciplinary methodology across natural and social sciences to examine relationships between biodiversity loss at sea and urban consumption with a focus on sea cucumber and dried seafood markets in New York City (NYC). The study identified 34 dried seafood retailers across three NYC Chinatown boroughs: Manhattan, Brooklyn, and Queens. Samples of sea cucumber were collected with Chinese-language labels indicating the commodity was from South America, a region of conservation concern. Comparison samples were taken from sea cucumbers labeled from Mexico and Japan. A mitochondrial DNA barcoding method was used to examine the taxonomic origin of 103 samples. Sequence data were successfully obtained from 74 of the samples, 8 of which were classified as brown sea cucumber (Isostichopus fuscus), an endangered species for which harvest is banned in several locations. Semi-structured interviews with dried seafood retailers and consumers (n = 64), moreover, revealed associations between consuming sea cucumber and enhancing human health and limited knowledge of product origins. Collectively, the findings reveal socio-ecological dynamics wherein endangered species on the market coupled with geographic market labeling practices and varying degrees of retailer and consumer knowledge negatively bear on marine biodiversity. Furthermore, given that brown sea cucumbers are abundant on the market, there is a need for developing genetic markers that can trace geographic origin to determine if species were legally harvested. These results indicate that more robust market labeling, training, genetic research, and public outreach are required to advance One Health in urban seafood markets.

Multiomic profiling of human clonal hematopoiesis reveals genotype and cell-specific inflammatory pathway activation

AbstractClonal hematopoiesis (CH) is an age-associated phenomenon that increases the risk of hematologic malignancy and cardiovascular disease. CH is thought to enhance disease risk through inflammation in the peripheral blood.1 Here, we profile peripheral blood gene expression in 66 968 single cells from a cohort of 17 patients with CH and 7 controls. Using a novel mitochondrial DNA barcoding approach, we were able to identify and separately compare mutant Tet methylcytosine dioxygenase 2 (TET2) and DNA methyltransferase 3A (DNMT3A) cells with nonmutant counterparts. We discovered the vast majority of mutated cells were in the myeloid compartment. Additionally, patients harboring DNMT3A and TET2 CH mutations possessed a proinflammatory profile in CD14+ monocytes through previously unrecognized pathways such as galectin and macrophage inhibitory factor. We also found that T cells from patients with CH, although mostly unmutated, had decreased expression of GTPase of the immunity associated protein genes, which are critical to T-cell development, suggesting that CH impairs T-cell function.

Phylogenetic insights into Central European Chorthippus and Pseudochorthippus (Orthoptera: Acrididae) species using ddRADseq data

The evolution of several orthopteran groups, especially within the grasshopper family Acrididae, remains poorly understood. This is particularly true for the subfamily Gomphocerinae, which comprises cryptic sympatric and syntopic species. Previous mitochondrial studies have highlighted major discrepancies between taxonomic and phylogenetic hypotheses, thereby emphasizing the necessity of genome-wide approaches.In this study, we employ double-digest restriction site-associated DNA sequencing (ddRADseq) to reconstruct the evolution of Central European Chorthippus and Pseudochorthippus species, especially C.smardai, P.tatrae and the C.biguttulus group.Our phylogenomic analyses recovered deep discordance with mitochondrial DNA barcoding, emphasizing its unreliability in Gomphocerinae grasshoppers. Specifically, our data robustly distinguished the C.biguttulus group and confirmed the distinctiveness of C.eisentrauti, also shedding light on its presence in the Berchtesgaden Alps. Moreover, our results support the reclassification of C.smardai to the genus Pseudochorthippus and of P.tatrae to the genus Chorthippus.Our study demonstrates the efficiency of high-throughput genomic methods such as RADseq without prior optimization to elucidate the complex evolution of grasshopper radiations with direct taxonomic implications. While RADseq has predominantly been utilized for population genomics and within-genus phylogenomics, its application extends to resolve relationships between deeply-diverged clades representative of distinct genera.

A DNA‐based evaluation of North American river otter diet with respect to fishes: Implications for the conservation of aquatic predators and their prey

Abstract The North American river otter Lontra canadensis was once widespread in North America but became extinct in the State of Indiana owing to habitat losses and unregulated trapping. However, following reintroduction, the population has expanded markedly since 1995, representing a notable conservation success story. The recent population growth has led to conflict with recreational angling groups who perceive otters as competitors. To investigate the extent of this competition, a DNA‐based study was conducted to determine whether material from the stomachs of trapper‐harvested otters could be used to identify fish prey species they had eaten. Representative specimens from 33 wild fish species were used as positive controls for DNA assays of otter prey. Mitochondrial DNA barcoding sequences generated from each fish species were used to develop a piscine barcoding assay to identify otter prey species. Fourteen otter carcasses were dissected, and multiple independent DNA extractions and mitochondrial sequencing reactions were performed on the prey bolus from each stomach. The sequences were analysed bioinformatically for taxonomic assignment to prey species. DNA evidence of nearly two dozen fish species, including species of recreational (e.g. bass) or conservation interest (darters) was found in the stomach material. This approach could help to determine the presence of prey species in otter diets, including those fish of conservation concern. That, in turn, would help to determine the best management options for conserving both otters and fish.

Genetic identification and gut content of Gerong fish Osteochilus waandersii from Larai River

Gerong fish, or Osteochilus waandersii, is a popular freshwater fish among the Jakun tribe in Kampung Peta, Johor National Park of EndauRompin, particularly for its use in traditional medicine to alleviate dizziness by consuming its raw gut. Despite its cultural significance, the genetic and feeding behavior of Gerong fish remains unclear. This study aimed to amplify the mitochondrial DNA barcode region of Gerong fish for genetic identification and to identify its gut content. The DNA barcodes were produced using polymerase chain reaction and single-pass DNA sequencing of partial COI gene, and the gut content was observed using stereo microscope. Several five partial COI gene sequences were successfully obtained after DNA sequencing. In addition, the results of BLAST analysis with percent identities of 96.24% to 97.20% along with the phylogenetic inference of maximum likelihood analyses suggested the Gerong fish as Osteochilus waandersii or the Waanders's hard-lipped barb. The sequences were deposited in GenBank (accession numbers MN223396-MN223400). Microscopic observation of the fish guts revealed undigested worms and digested aquatic mosses, known as lumut batuk. These findings mark a significant step forward in understanding the genetics and feeding behavior of the Gerong fish.

Molecular characterization of mitochondrial COI gene sequences in Micraspis allardi from Pakistan.

The Coccinellidae is a highly diversified family of order Coleoptera. Coccinellid ladybirds are well known for their role as biological control agent against varied range of agricultural pests. The samples of coccinellid ladybird collected from Pakistan were identified and characterized as Micraspis allardi (Mulsant, 1866). This is one of the least-studied ladybird species with limited work on its ecological distribution as a biological control agent. The genus Micraspis has vast genetic diversity with a possible presence of unknown number of cryptic species. Sequence information of some species of the genus Micraspis are present in NCBI database. However, least molecular data or sequences describing M. allardi could be available from database. Therefore, morphological and molecular characterization was imperative for this species. Here, the samples collected from sugarcane field of Faisalabad District of Pakistan and were identified by using morphological and molecular protocols. For molecular identification, two different regions of mitochondrial cytochrome c oxidase I (COI) gene (COI-5' and COI- 3') were used as molecular markers for the identification of the species. Morphological appearance, DNA sequence similarity searches and phylogenetic analysis collectively indicated it as M. allardi. To the best of our knowledge, this is the first report providing molecular evidence of M. allardi using mitochondrial DNA barcode region (658bp) as well as mtCOI-3' sequences (817bp). The study will help in understanding population genetics through diversity analysis, ecological role, and phenotypic structures associated with the geographic range of this species.

MtDNA data reveal disparate population structures and High Arctic colonization patterns in three intertidal invertebrates with contrasting life history traits

IntroductionPost-glacial climate variation is known to have influenced the distribution of marine species in the North Atlantic. In particular, the Atlantic side of the Arctic has experienced strong fluctuations in both atmospheric and sea surface temperature, as well as seasonal ice coverage since the last glacial maximum (LGM). Here, we aim to unveil the phylogeography and historical demography of three rocky intertidal marine invertebrates showing a trans-Atlantic distribution and presently inhabiting the Arctic: Gammarus oceanicus, Littorina saxatilis and Semibalanus balanoides.MethodsWe used a large amount of mitochondrial DNA barcode data, both newly-obtained and stored in public databases. We performed phylogeographic and demographic analyses on 1119 G. oceanicus, 205 L. saxatilis, and 884 S. balanoides sequences.ResultsOur results show that all three of these boreal species have expanded their effective population sizes in the high Arctic Svalbard Archipelago since the LGM. Analyses investigating the origin of all these populations point to the eastern Atlantic.DiscussionBased on our results we conclude that the expansion of these boreal species to the Arctic possibly happened during an earlier warm cycle of the Holocene era, and is probably not the result of the recent ‘Atlantification’ of the Arctic. We also discuss the effects of dispersal potential on population structure as an important aspect of comparative biogeographical studies.

Delimiting phylogeographic diversity in the genomic era: application to an Iberian endemic frog

Abstract The rich genetic and phenotypic diversity of species complexes is best recognized through formal taxonomic naming, but one must first assess the evolutionary history of phylogeographic lineages to identify and delimit candidate taxa. Using genomic markers, mitochondrial DNA barcoding, and morphometric analyses, we examined lineage diversity and distribution in the Iberian endemic frog Rana parvipalmata. We confirmed two deep phylogeographic lineages, one relatively homogenous genetically, found in Asturias and adjacent areas (T2), and one more fragmented and locally genetically impoverished, restricted to Galicia (T1). Analyses of their hybrid zone suggested a shallow transition characterized by far-ranging admixture, which was modelled by a wide geographic cline (~60 km for the genome average) and no obvious barrier loci (i.e. loci showing disproportionally restricted introgression). The relatively young T1 and T2 have thus remained reproductively compatible, which argues against their distinction as biological species, and we accordingly describe T2 as a new subspecies, Rana parvipalmata asturiensis ssp. nov. Remarkably, we highlight striking discordances between mitochondrial and nuclear distributions across their hybrid zones, as well as between their genetic and morphological differentiation. Our study illustrates how genomic-based phylogeographic frameworks can help decipher the high genetic and phenotypic variation of species complexes and substantiate the taxonomic assessment of candidate lineages.

Phylogenetic Structure Revealed through Combining DNA Barcodes with Multi-Gene Data for Agrodiaetus Blue Butterflies (Lepidoptera, Lycaenidae).

The need for multi-gene analysis in evolutionary and taxonomic studies is generally accepted. However, the sequencing of multiple genes is not always possible. For various reasons, short mitochondrial DNA barcodes are the only source of molecular information for some species in many genera, although multi-locus data are available for other species of the same genera. In particular, such situation exists in the species-rich butterfly subgenus Polyommatus (Agrodiaetus). Here, we analyzed the partitioning of this subgenus into species groups by using three data sets. The first data set was represented by short mitochondrial DNA barcodes for all analyzed samples. The second and third data sets were represented by a combination of short mitochondrial DNA barcodes for part of the taxa with longer mitochondrial sequences COI + tRNA-Leu + COII (data set 2) and with longer mitochondrial COI + tRNA-Leu + COII and nuclear 5.8S rDNA + ITS2 + 28S rDNA sequences (data set 3) for the remaining species. We showed that the DNA barcoding approach (data set 1) failed to reveal the phylogenetic structure, resulting in numerous polytomies in the tree obtained. Combined analysis of the mitochondrial and nuclear sequences (data sets 2 and 3) revealed the species groups and the position within these species groups, even for the taxa for which only short DNA barcodes were available.

Performance of DNA barcodes for informing the subspecies controversy in North American populations of Callophrys gryneus (Hübner, [1819]) (Lepidoptera: Lycaenidae)

Abstract Analysis of genetic divergence in published mitochondrial DNA barcodes from ‘juniper’ and ‘cedar’ hairstreaks, Callophrys gryneus (Hübner) and Callophrys muiri (Hy. Edwards), together with a comparison of phenotypic differences in populations from North America, have been used to address the long-standing controversy regarding subspecies and species assignments in this confusing group of hairstreaks. The most recent view holds that C. gryneus is a single polytypic species comprising 11 nominal subspecies and that C. muiri from California is a valid species. An alternative view subdivides C. gryneus into three nominal species, C. gryneus (s.s.), Callophrys siva and Callophrys nelsoni, each with several named subspecies, and retains C. muiri as a full species. Haplotype networks, phylogenetic analysis and analysis of molecular variance of barcodes provided evidence supporting the subspecies scenario. However, barcodes from closely related congeners of C. gryneus (s.l.) revealed pairwise genetic divergences that would also support elevating three subspecies (C. g. castalis, C. g. siva and C. g. nelsoni) to full species. The barcode data also raise the question of whether C. muiri warrants full species status. Larval host plant preference as a possible driver of genetic differentiation in western populations of this group is also discussed briefly.

Uncovering the diversity of rodents and shrews in the Chebera Churchura National Park, Ethiopia

Abstract The recently established Chebera Churchura National Park represents a mosaic of savanna and montane evergreen forest. A report on small mammals (2013) showed their relatively high diversity, but species identification was based only on external morphology. We conducted a new field survey and genetic analyses to shed light on diversity and biogeographical affinities of small mammals in this under-explored national park. Based on results of mitochondrial DNA barcoding, 14 species of rodents and two species of shrews, including Crocidura turba (new to the Ethiopian fauna), were identified. Moreover, a new taxon of gerbils – presumably representing a new species – of the genus Gerbilliscus was discovered. We show that forest and savanna species often inhabit the same localities, which can be explained by the mosaic nature of biotopes. These results document the uniqueness of Chebera Churchura NP, especially in comparison with other Ethiopian national parks, and argue for its protection and further development.