Search Results (109)

The Asian larch bark beetle, I. subelongatus Motschulsky, is a severe pest of various Larix species in its natural range. This study reports the degree of similarity among cuticular hydrocarbon (CHC) profiles of six populations of I. subelongatus in northeastern China. Thirty individual or isomeric mixtures of hydrocarbons were identified by solid-phase microextraction (SPME) coupled with gas chromatography–mass spectrometry (GC–MS). The hydrocarbon components consist of straight-chain alkanes, alkenes, and methyl-branched hydrocarbons with carbon chain lengths ranging from 24 to 31. Among these, four CHCs (n-C₂₅, 9-C_27:1, n-C₂₇, and 3-meC₂₇) can serve as chemotaxonomic markers to identify I. subelongatus. No qualitative differences were detected between males and females in any of the six populations, but significant quantitative differences were observed in some CHCs. Phylogenetic analysis based on CHC profiles showed only minor differences compared to analysis based on partial mtDNA COI sequences regarding bark beetle species affinity. These results establish a rapid chemotaxonomic method and provide a basis for further investigations into the functions of CHCs in I. subelongatus. Full article

Background/Objectives: Fish are vital in the Mediterranean diet, offering protein, nutrients, and ω-3 fatty acids. Greek consumers favor wild-caught, high-value fish like the dusky grouper (Epinephelus marginatus) classified as “vulnerable” and the white grouper (Epinephelus aeneus) classified as “near threatened” species, according to the IUCN Red List. Due to their premium prices and complex supply chains, these species are susceptible to fraud, especially through mislabeling. This practice not only deceives consumers but also poses health risks and encourages illegal fishing. DNA-based methods have shown effectiveness in accurately identifying species, even in processed samples. The aim of this study is to apply high-resolution melting analysis (HRM) as a rapid, effective method for monitoring the appropriate labeling of the two Epinephelus species in the Greek market. Methods: In this study, fresh fish from Greek catches as well as cooked, frozen, and filleted samples collected from the Greek market were identified using DNA barcoding. HRM analysis based on single nucleotide polymorphisms (SNPs) was used to differentiate between locally sourced E. marginatus and E. aeneus from their imported counterparts or from other species available in the Greek market that could be used in substitution incidents. Results: Using HRM analysis, cases of species mislabeling were identified and were also confirmed using sequencing. Conclusions: HRM analysis proved to be an accurate and cost-effective method for rapidly processing a large number of samples; therefore, it could serve as a valuable tool in extensive market controls as well as for bio-diversity conservation monitoring. Full article

The Danube Clouded Yellow (Colias myrmidone) has experienced one of the most dramatic declines among European butterflies. To estimate genetic diversity in the last population in Poland that has survived in the Knyszyn Forest (KF), we analyzed mitochondrial (COI) and nuclear (EF-1α) polymorphisms in individuals sampled in 2014 and 2022. The results were compared with genetic data obtained in 2014 from a recently extirpated nearby population (Czerwony Bór, CB). Because mtDNA polymorphisms in insects can be modulated by endosymbionts, the samples were screened for Wolbachia. The polymorphism of EF-1α indicated that diversity was gradually decreasing. The KF experienced rapid demographic processes, manifested by a significant change in allele frequency. The small differentiation in nuclear markers between the KF and CB in 2014 suggests that the regional population used to be genetically uniform. Four COI haplotypes that were identified in this study probably belong to two different haplogroups. Wolbachia was detected only in individuals with one specific haplotype, and the prevalence was female-biased, suggesting the induction of two reproductive manipulations. The most common COI haplotype found in Poland was the same as that reported from other parts of Europe, not only for C. myrmidone but also C. caucasica. These results allow us to question the distinctiveness of each taxa. Full article

Honey is a widely consumed food product frequently subjected to adulteration, with the mislabeling of its botanical or geographical origin being one of the most common practices. Determining the entomological origin of honey is particularly challenging but of high relevance for ensuring its authenticity, especially for products with protected designation of origin (PDO) labels. This study presents a novel DNA metabarcoding approach targeting a highly informative 406 bp fragment of the cytochrome c oxidase I (COI) gene to differentiate among the three major mitochondrial lineages (A, M, and C) of honey bees (Apis mellifera L.) native to Europe. The target region was selected based on the calculated fixation index (F_ST), which is frequently used in Population Genetics as a measure of differentiation between populations. The approach was validated with 11 honey samples of known entomological origin and applied to 44 commercial honeys from 13 countries. The approach demonstrated high sensitivity, accurately identifying the entomological origin of honey, including samples produced by honey bees of varying ancestries, which could not be resolved by previous methods based on real-time PCR coupled with high-resolution melting (PCR-HRM) analysis. The results demonstrate the effectiveness of COI metabarcoding in verifying honey authenticity and highlight the predominance of C-lineage honey bees in the production of commercial honeys from northwestern Europe. This finding suggests a limited presence of the native M-lineage ancestry, underscoring the need for conservation efforts. Full article

Lance nematodes (Hoplolaimus spp.) feed on the roots of various plants, including key horticultural products. An investigation of the genetic diversity and structure of six Hoplolaimus species, utilizing the cytochrome c oxidase I (COI) of the mtDNA gene, was based on 174 sequences available on the NCBI. Based on the COI of mtDNA, the haplotype analysis revealed 44 haplotypes. Nucleotide diversity was low among all species of Hoplolaimus, except for H. magnistylus (π = 0.04915) and H. stephanus (π = 0.06746). In contrast, haplotype diversity (Hd) was high, especially for H. stephanus (Hd = 0.89) and H. pararobustus (Hd = 0.90). Phylogenetic analysis grouped the various populations into eight clades, and the result showed that H. magnistylus was placed in three different clades, which showed high variability in haplotype supported by the haplotype network. Neutrality tests and mismatch distribution based on the mtDNA supported the hypothesis of a constant population with no expansion in Hoplolaimus, except for H. concaudajuvencus (Tajima (D) = −0.84971) and H. columbus (Tajima (D) = −0.87674). In conclusion, genetic analysis showed a neutral evolution amongst the Hoplolaimus species. The result of the present study provides a better insight into the Hoplolaimus species toward species delimitation and managing this plant-parasitic nematode in various crops. Full article

The trans-Palearctic moth Phyllonorycter populifoliella (Lepidoptera: Gracillariidae) is a major pest of the North American Populus balsamifera and its hybrids widely planted as ornamentals in North Asia (i.e., the Asian part of Russia). We DNA barcoded Ph. populifoliella from distant geographical populations in Russia and analyzed them together with the data from eight European countries and India to estimate intraspecific variability and the haplotype richness in the Palearctic, and specifically in North Asia. Furthermore, using next-generation sequencing (NGS, Sequel platform, PacBio), we investigated larval and pupal remnants found in an old herbarium from the Nearctic, where P. balsamifera occurs naturally, to verify if any events of the moth introduction to this biogeographic zone happened in the past. Relatively high intraspecific variability in the COI gene of mtDNA, reaching 3.73%, was recorded in Ph. populifoliella. Overall, 30 COI haplotypes were defined in 83 specimens from the Palearctic, with a noticeable richness in North Asia (21 haplotypes). Using NGS, the remnants of 14 Phyllonorycter specimens dissected from up to 174-year-old herbaria from the Palearctic and Nearctic were sequenced, and four moth species were identified. Among them, there were three Palearctic species, Ph. populifoliella, Ph. pastorella (Zeller), and Ph. apparella (Herrich-Schäffer), and one Nearctic, Ph. nipigon (Freeman). No evidence of Ph. populifoliella introduction to North America was documented based on the examination of the herbarium dated 1850–1974. Three specimens of Ph. populifoliella identified from herbaria from Austria and Poland (dated 1879–1931) represented one haplotype (H7) known from the recent time. Overall, our study clarifies the modern range, provides insights into phylogeography, and defines the haplotype richness of the native leafminer outbreaking on the alien host. Furthermore, it underlines the use of old herbaria to explore the historical distribution of endophagous insect species. Full article

Due to the complexity of taxonomic classification based on the classical morphological characters of copepods, phylogenies have been ambiguous. In this study, we investigate the phylogeny of copepods, including four species from three orders, in the saltern of Sfax using the small subunit of nuclear ribosomal RNA genes (18S). In the studied area, copepods seemed to be a polyphyletic group, and the genetic structure of these crustaceans is complex and problematic. We have also used two mitochondrial markers, the cytochrome c oxidase subunit I (mtCOI) gene and the cytochrome b (Cytb) sequence data, in order to investigate the genetic diversity and differentiation in a total of 96 individuals from two sets of Paracartia grani, sampled from two ponds with different salinities (42 PSU and 61 PSU). All of the results presented here suggest a low genetic diversity among P. grani species and a weak genetic structure between the sets. The nucleotide and haplotype diversity of P. grani were extremely low, indicating the homogeneity of the two sets, which could be combined into one set living in different ranges of salinity. This small genetic diversity is possibly due to the confined natural distribution range and strong selective pressure in a saltern environment. These data also suggest that gene flow is the main factor shaping the genetic structure of the studied sets. Full article

Chrysodeixis includens and Rachiplusia nu are two species belonging to the Plusiinae subfamily within the Noctuidae family. Due to their morphological similarity, the identification of their larvae is difficult and time-consuming. A rapid and accurate identification of these two species is essential for their management as these species exhibit differential susceptibilities to insecticides and crops engineered to express Bacillus thuringiensis (Bt) proteins, and a molecular tool can easily provide this differentiation. Currently, molecular analysis can identify these species through genetic sequencing, an expensive and time-consuming process. In our study, after sequencing part of the mtDNA cytochrome c oxidase I (COI) gene and based on the differences found in the gene of each species, a set of species-specific primers was developed: one reverse primer common to both species and two forward primers, specific to each species, amplifying fragments of 199 base pairs (bp) for C. includens and 299 bp for R. nu. Our results indicate that the primers were specific for these species, enabling the identification of individuals directly through agarose gel. The new methodology proved to be accurate, rapid, and reliable for the correct identification of these two species of loopers. Full article

The present study describes two new species of monopisthocotylans parasitizing the gills of Cyphocharax modestus (Characiformes: Curimatidae) from two localities of the Upper Paraná River basin, state of São Paulo, Brazil. Urocleidoides saghirus n. sp. and Urocleidoides taquariensis n. sp. differ from other congeners mainly in the morphology of the accessory piece: comprising two subunits (one scythe-shaped subunit, and one adjacent piece with a narrow base broadening towards the upper end, which is axe-shaped) in Urocleidoides saghirus n. sp., and a straight rod with a bifurcated proximal portion, a distal portion ending in a point, and two curved handles arising from the medial portion and uniting close to the distal portion in Urocleidoides taquariensis n. sp. The phylogenetic analyses revealed Urocleidoides to be non-monophyletic, indicating the need for a taxonomic review. This study provides the first molecular sequences for Urocleidoides spp. parasitizing Curimatidae, including the newly described species. These findings revealed distinct clades and suggested possible host specificity, underscoring the importance of additional research to better understand the phylogenetic relationships within the genus. Full article

The fauna of Lepidoptera in the Mediterranean is still inadequately documented. As a result, even remotely complete DNA barcode libraries (mt. COI (cytochrome c oxidase 1) gene) are lacking in most areas. This proposed gap is being analyzed for the first time for the fauna of North Cyprus. In the initial phase, 248 morphospecies from 29 families (exclusive Heterocera) were sampled, sequenced and compared with existing DNA reference sequences in the global BOLD database (Barcode of Life Data Systems) via BINs (Barcode Index Numbers). A total of 194 species could be unequivocally assigned to a Linnaean taxon. Additionally, six species previously unidentified in BOLD, as well as fourteen species without reference barcodes, were identified at the species level. Twenty-four of these species were new records for Cyprus. In addition, 25 taxa with new BINs could not be assigned to a valid species due to potential cryptic diversity or the lack of relevant revisions. Furthermore, a few species could not be identified due to barcode sharing and/or potential misidentifications in BOLD. Overall, approximately 20% of the samples could not be identified using the existing DNA barcode libraries, a significant deficit for European standards, which should be addressed as a priority issue in future studies. Full article

A partial mitochondrial DNA Cytochrome Oxidase subunit I (mtCOI) gene haplotype variant of the coconut rhinoceros beetle (CRB) Oryctes rhinoceros, classed as ‘CRB-G (clade I)’, has been the focus of much research since 2007, with reports of invasions into new Pacific Island locations (e.g., Guam, Hawaii, Solomons Islands). For numerous invasive species, inference of invasion biology via whole genome is superior to assessments via the partial mtCOI gene. Here, we explore CRB draft mitochondrial genomes (mitogenomes) from historical and recent collections, with assessment focused on individuals associated within the CRB-G (clade I) classification. We found that all Guam CRB individuals possessed the same mitogenome across all 13 protein-coding genes and differed from individuals collected elsewhere, including ‘non-Guam’ individuals designated as CRB-G (clade I) by partial mtCOI assessment. Two alternative ATP6 and COIII partial gene primer sets were developed to enable distinction between CRB individuals from Guam that classed within the CRB-G (clade I) haplotype grouping and CRB-G (Clade I) individuals collected elsewhere. Phylogenetic analyses based on concatenated ATP6–COIII genes showed that only Guam CRB-G (clade I) individuals clustered together, and therefore Guam was not the source of the CRB that invaded the other locations in the Pacific assessed in this study. The use of the mtCOI and/or mtCOIII genes for initial molecular diagnosis of CRB remained crucial, and assessment of more native CRB populations will further advance our ability to identify the provenance of CRB invasions being reported within the Pacific and elsewhere. Full article

Heterakis spumosa, a parasitic worm infecting rodents, is globally prevalent in black rats, brown rats, and house mice. It is hypothesized to originate from Asia due to its widespread presence in Southeast Asia in various Murinae. Previous molecular studies focused on European, African, and Japanese specimens, but none included samples from the putative native range. Rodents were collected between 2008 and 2015 across various localities in Southeast Asia and Europe, identified by morphology or genetic barcoding. Viscera were examined or preserved for later inspection. DNA was extracted from H. spumosa. PCR amplification targeting the mtCOI gene and ITS1 region was conducted in this study using newly designed primers (based on Heterakis reference sequences). PCR amplicons were subsequently sequenced and analyzed. In this study, the phylogenetic analysis using ITS1 sequences revealed that Heterakis samples from Thai and Laotian rodents belong to the species H. spumosa, exhibiting low genetic variation compared to samples from other regions. Genetic distance calculations using mtCOI sequences confirmed the marked distinction of H. spumosa from other Heterakis species. Our phylogenetic analyses using partial mtCOI and ITS1 sequences have significantly enhanced our comprehension of the genetic diversity and evolutionary history of the nematode H. spumosa. Full article

The taxonomic status of many species of the family Cyclocoelidae is still unclear. Two species of cyclocoelids, Harrahium obscurum and Morishitium dollfusi, were collected from the air sacs of birds (Turdus merula and Tringa ochropus) inhabiting the Middle Volga region (European Russia). Here, we provide the first detailed morphological description of these cyclocoelids and combine it with the first molecular phylogenetic analysis of Cyclocoelidae from birds in Russia based on partial sequences of their 28s rDNA and coI mtDNA genes. Specimens of both flatworm species from different host individuals differ slightly in body shape and size, which probably reflects host-induced intraspecific variability. For the first time, we have shown that a stable morphological character such as the length of the vitelline fields in the studied digeneans is variable at the species level and cannot be used in their morphological diagnosis. Full article

The mitochondrial cytochrome c oxidase subunit I (COI) genes of six endangered goose breeds (Xupu, Yangjiang, Yan, Wuzong, Baizi, and Lingxian) were sequenced and compared to assess the genetic diversity of endangered goose breeds. By constructing phylogenetic trees and evolutionary maps of genetic relationships, the affinities and degrees of genetic variations among the six different breeds were revealed. A total of 92 polymorphic sites were detected in the 741 bp sequence of the mtDNA COI gene after shear correction, and the GC content of the processed sequence (51.11%) was higher than that of the AT content (48.89%). The polymorphic loci within the populations of five of the six breeds (Xupu, Yangjiang, Yan, Baizi, and Lingxian) were more than 10, the haplotype diversity > 0.5, and the nucleotide diversity (Pi) > 0.005, with the Baizi geese being the exception. A total of 35 haplotypes were detected based on nucleotide variation among sequences, and the goose breed haplotypes showed a central star-shaped dispersion; the FST values were −0.03781 to 0.02645, The greatest genetic differentiation (FST = 0.02645) was observed in Yan and Wuzong breeds. The most frequent genetic exchange (Nm > 15.00) was between the Wuzong and Yangjiang geese. An analysis of molecular variance showed that the population genetic variation mainly came from within the population; the base mismatch differential distribution analysis of the goose breeds and the Tajima’s D and Fu’s Fs neutral detection of the historical occurrence dynamics of their populations were negative (p > 0.10). The distribution curve of the base mismatches showed a multimodal peak, which indicated that the population tended to be stabilised. These results provide important genetic information for the conservation and management of endangered goose breeds and a scientific basis for the development of effective conservation strategies. Full article

The two cryptic grasshopper species of the genus Stenobothrus, S. eurasius and S. hyalosuperficies, demonstrate different acoustic behavior despite a strong similarity in morphology. A hybridization between these species is possible in the contact zone; however, there are little molecular data about the relationships of these species. The analysis of the mtDNA control region (CR) reveals that haplotypes of S. hyalosuperficies have more in common with the more distant Stenobothrus species than with the closely related S. eurasius. In the contact zone, S. eurasius has mt-haplotypes shared with S. hyalosuperficies, which might indicate an introgression of mtDNA from S. hyalosuperficies to the S. eurasius gene pool. We also analyze the structure and evolutionary rate of the mtDNA CR for the Stenobothrus genus and estimate the time of divergence of the species within the genus. The phylogenetic tree of the tribe Stenobothrini reconstructed with either the CR or COI gave the same four groups. The phylogenetic tree of the Stenobothrus genus has a star-like topology with each mtDNA haplotype found in any analyzed species, except S. eurasius, which forms a separate branch. The maximum degree of incomplete lineage sorting can demonstrate either ancestral polymorphism or introgression. Full article