The genus Spauligodon Skrjabin, Schikhobalova, Lagodovskaja, 1960 has a wide global distribution and comprises 52 species, of which 12 are reported in the Neotropical Region, parasitizing mostly reptiles. In Brazil, there are records of only one species, namely Spauligodon oxkutzcabiensis (Chitwood, 1938), which is an abundant parasite in lizards of the family Phyllodactylidae. The original description of S. oxkutzcabiensis was based on the analysis of a restricted set of morphological and morphometric characteristics, with limited taxonomic detail. This study provides a detailed redescription of S. oxkutzcabiensis, including the first observations using scanning electron microscopy (SEM), as well as the first genetic characterization of the species. During a parasitological survey carried out in May 2024, in the municipality of Mauriti, State of Ceará, Brazil, specimens of Spauliodon were found in the intestine of Phyllopezus periosus (Squamata: Phyllodactylidae). The nematodes were identified as S. oxkutzcabiensis based on the presence of well-developed lateral wings in males, three pairs of papillae in the posterior region, with the last pair not supported by the caudal wing, and the presence and number of spines on the tail of female specimens. The nematodes were characterized morphologically by light microscopy and SEM. The parasites were also subjected to molecular analysis, with the amplification and sequencing of partial fragments of the 18S and 28S rDNA genes. Based on the analysis, it was possible to provide a more comprehensive morphological description, such as the location of the nerve ring, males with a tripartite lateral wing in the region where it begins in the esophagus, and females with an elongated muscular ovijector, characteristics not observed before. This improves the specific diagnosis of S. oxkutzcabiensis (Chitwood, 1938), which was supplemented by molecular data, contributing to knowledge about species variability and the general taxonomy of members of the genus Spauligodon in the Neotropical region.

Morphotaxonomy of immature stages can resolve misidentification, proliferation of cryptic species, and timely and effective management of vector species of Culicoides Latreille (Diptera: Ceratopogonidae) at the onset of development. Among 84 valid Culicoides species reported from India, many are unknown to be immature, and some are partially described. Culicoides jacobsoni Macfie and Culicoides oxystoma Kieffer are two species considered to be proven vectors of the bluetongue virus. In the present investigation, Culicoides thurmanae Wirth and Hubert were first reported in India. The surface structures of eggs of C. jacobsoni and C. thurmanae are newly described and photomicrographed using Scanning Electron Microscopy (SEM). The gravid females were collected using a UV LED light trap and identified morphologically. The eggs were retrieved by dissecting their abdomens and processing them under SEM. Simultaneously, the laboratory-reared oviposited and hatched eggs of C. oxystoma are also described. The field-collected engorged females were transferred onto moistened cotton beds for oviposition. Embryonated eggs were then transferred into pre-prepared rearing plates consisting of cattle manure, yeast solution, and nutrient broth solution, allowing for successive egg hatching and rearing of larval instars. The larval instar differentiation and morphometric measurements of four larval instars were newly noted. This investigation also highlights pupal diagnoses across sexes.

The ability to delimit species and clades is essential to understand their ecology, evolutionary history, and asses their conservation need. Gymnorhamphichthys, known as Sand knifefishes, is a genus of South American weakly electric fishes. The clade has five valid species, most of which have broad geographic distributions with substantial phenotypic variation. These variations resulted in a convoluted taxonomic history, lack of clear diagnostic features and misidentification. Since older taxonomic methodologies failed to distinguish between possible pseudo cryptic species, we expect newer methodologies such as micro-computed tomography, 3D morphometrics and multi loci molecular sequences to allow us to elucidate patterns of diversity within this clade. Data was collected from specimens in museums across North America (ANSP, FMNH, UF, USNM), Brazil (INPA, MZUSP, MNRJ, DZUFMG, LBP, PUC-Minas) and Europe (MHNG). Non-destructive microtomography allowed for character search, osteological description, and species delimitation. Phylogenetic tree was created combining both molecular and morphological matrix following maximum likelihood and posterior Bayesian analysis. Preliminary results support the five valid species already described with external and internal characters, molecular data, and ecological traits. Two possible new species from Guyana and Trombetas region are being evaluated. Results from this research will clarify the alpha taxonomy of Gymnorhamphichthys, addressing a major gap in the field of neotropical freshwater fishes and permit future studies such as how intraspecific variation can be transformed into interspecific differences, evolutionary and biogeography trends of Neotropical freshwater fishes and morphological evolution.

Introduction: Accurate and rapid taxonomic classification of microbial genomes is fundamental for microbiology, epidemiology, and biodiversity monitoring. Existing bioinformatic pipelines often require large computational resources and multiple manual steps. Objective: We developed GenoScanner, a shell-based, high-speed, and hypothesis-driven pipeline to classify microbial genomes within minutes, integrating genomic analysis, phylogenetic inference, and molecular speciation testing under the Phylophenetic Species Concept. Methods: The pipeline compares the investigated genome against a curated genomic database (nomenclaturally valid genomes) to identify the closest phylogenetic neighbors. Subsequent Average Nucleotide Identity (ANI) estimation and robust phylogenetic reconstruction was carried out with JolyTree; Poisson Tree Processes Models (bPTP default or mPTP optional) were used to provide the basis for Bayesian species delimitation testing. This software relies on well-established bioinformatic tools such as NCBI Entrez Direct, BLAST+, and Biopython, among others. The rationale of GenoScanner explicitly evaluates three hypotheses to achieve better taxonomic resolution: 1. Genomic Coherence—via mutational distances and nucleotide identity; 2. Phylogenetic Monophyly—through rapid full-genome tree estimation; 3. Molecular Speciation—under probabilistic PTP models. Results & Advantages: The proposed framework enables rapid genome classification within minutes, significantly accelerating taxonomic workflows. By integrating genome-based phenetic data like ANI and Mash distance, phylogenomic analysis, and speciation-level evidence, it enhances the resolution and reliability of species assignments. Species delimitation is performed with robust accuracy using the PTP models, and full reproducibility is ensured through a standardized genomic databases. The approach is particularly effective for classifying bacteria, fungi, algae, and protists that are well represented in genomic repositories. These features collectively support diverse applications in molecular systematics, epidemiological monitoring, and biodiversity assessment. Conclusion: GenoScanner provides an efficient, reproducible, and conceptually robust framework for genome-based microbial taxonomy. By combining speed, accuracy, and formal species delimitation testing, it serves as a practical tool for both research and surveillance applications. Availability: https://github.com/ayixon/genomescanner.

The genus Ganoderma, belonging to the family Ganodermataceae, comprises a group of basidiomycete fungi with wide geographic distribution and scientific interest. Despite its ecological relevance and biotechnological potential, its taxonomy remains complex due to high morphological variability among species, influenced by environmental and nutritional factors that directly affect phenotypic characteristics. This study aimed to perform morphological and molecular characterization of eight fungal isolates of Ganoderma to support taxonomic identification and provide insights into their phylogenetic relationships. The eight isolates evaluated were cultured in PDA medium, morphologically characterized, and subjected to microscopic observation of hyphal structures. Mycelial growth was evaluated in different media (MEA, PDA, and SDA) and temperatures (18°C, 28°C, and 38°C) to understand the isolates physiological behavior. Subsequently, genomic DNA extraction was performed, according to the protocol by Vicente et al. (2008) and sequencing of the ITS region of ribosomal DNA, used as a marker for molecular fungi identification. Microscopically, all strains presented the trimitic hyphal system, formed by generative, skeletal, and ligative hyphae. These structures, together with colony macromorphology, confirmed generic identity. However, species-level identification was only possible through molecular analysis, which revealed four strains of G. lucidum, one of G. sessile, one of G. lingzhi, and one with ambiguous identity between G. resinaceum and G. sessile. Physiological analysis showed that medium composition directly influenced development, with some combinations promoting mycelial expansion. The temperature of 28°C favored mycelial growth in most isolates, especially G. lucidum and G. lingzhi, indicating species-specific physiological profiles. These variations reveal the specific physiological profile of each species. These data show that the traditional approach, based on morphology, is limited in delimiting Ganoderma species. The integration of microscopic characteristics, and molecular data, is essential to ensure accurate identification.

Thermal springs represent extreme ecological niches characterized by elevated temperatures and mineral-rich waters, shaping highly specialized microbial communities. Cyanobacteria, as primary producers in these habitats, exhibit remarkable adaptations, yet their diversity and phylogenetic relationships in Serbian geothermal systems remain poorly documented. In this study, we applied a molecular systematics approach based on 16S rRNA gene sequencing to characterize cyanobacterial assemblages from five thermomineral springs in Serbia. Field sampling was conducted during October and November, and all strains were subsequently isolated and cultivated in BG11 medium for several months to ensure stability and purity prior to molecular analysis. The molecular analyses also facilitated the confirmation of morphological identification. Leptolyngbya sp. (WUC607) was detected in Niška Banja (38–39 °C), while Leptolyngbya sp. (CALU1715) was found in Lopatnica (≈29 °C) and Šarbanovac (30 °C). Wilmottia sp. (CCNU0051) was identified in Brestovačka Banja (35 °C). In Vranjska Banja (surface 93 °C), both Nodosilinea sp. (PGN35) and Anabaena variabilis ("Vasse River Type 3") were found. This study provides molecular characterization of cyanobacterial communities from Serbian thermal springs, enriching a knowledge in thermophilic microorganisms and potentially discovery of new species. The findings emphasize the value of molecular systematics for better understanding evolutionary adaptations to thermomineral environments. Moreover, these thermophilic lineages represent potential candidates for further research on thermal tolerance mechanisms and potential biotechnological applications, such as bioactive compound production.

This study aimed to investigate the diversity of nematodes exhibiting various feeding strategies—including fungivorous, bacterivorous, predatory, and insect-associated forms—particularly those linked with bark beetles and Dutch elm disease. Between 2021 and 2023, eighty samples of dried elm bark and associated beetles were collected from Golestan Forest Park, Iran. Nematodes were extracted, fixed, and mounted on permanent microscope slides for morphological examination. Identification was based on morphological and morphometric analyses using standard keys, original species descriptions, and, where needed, molecular data. Fourteen species from ten genera belonging to the superfamilies Aphelenchoidea and Sphaerularioidea were identified. Among them, one population of Laimaphelenchus sp. exhibited a unique combination of morphological characters not found in any previously described species and is therefore proposed as a new species. Key diagnostic features include a body length of 336–446 μm, stylet length of 7–8.5 μm, three lateral lines, an elongated rectangular spermatheca containing sperm, absence of a vulval flap, and a posterior uterine sac measuring 3–4 times the vulval body width. The semi-cylindrical tail terminates in a flat disc with a stalk-like extension bearing four branched tubercles, each ending in a saucer-like structure with fringed, finger-like appendages. Males are present and possess spicules with a short, straight condylus and a blunt-tipped conical rostrum. The combination of these features places the species near L. patulus, L. australis, L. phloesini, and L. vescus, but distinct morphological differences support its recognition as a novel taxon within the genus Laimaphelenchus.

Stonefishes (Family: Synanceiidae) are venomous, benthic marine fishes with cryptic morphology and unresolved taxonomy. Despite their ecological and medical significance, they are poorly documented in Indian waters, particularly along the Kerala coast. This study employed an integrative approach, combining morphological and mitochondrial DNA analyses, to clarify species boundaries. Specimens were collected as bycatch from trawl landings at Munambam, Kalamukku, and Sakthikulangara (January–April 2025). Forty-seven morphometric measurements and five meristic counts were recorded per specimen. Mitochondrial DNA was extracted from muscle tissue, and the cytochrome c oxidase subunit I (COI) gene was amplified via PCR. Sequences were aligned, and phylogenetic relationships were inferred using the Maximum Likelihood method. Eight species were identified—Apistus carinatus, Inimicus didactylus, I. sinensis, Choridactylus multibarbus, Minous monodactylus, M. inermis, M. trachycephalus, and M. dempsterae. Morphological identifications were confirmed through DNA barcoding, with COI-based phylogeny revealing eight distinct clades and interspecific genetic divergence of 4.1–24.6% (mean: 19%). Distinct length–weight relationships were established for M. inermis and M. dempsterae. This first integrative taxonomic account of Synanceiidae from the Kerala coast demonstrates the value of combining classical and molecular tools for delineating cryptic marine species and provides a foundation for accurate biodiversity assessment, ecological monitoring, and conservation planning.

Ciliates are ecologically important microeukaryotes that transfer energy, remineralise nutrients, and regulate microbial communities. Their sensitivity to environmental changes makes them reliable bioindicators of the state of marine ecosystems. Although their taxonomy and ecology are well documented, conventional monitoring methods often overlook cryptic and rare species. Integrative approaches combining morphological and molecular techniques are therefore essential to capture the full spectrum of ciliate diversity. In this study, a dual taxonomic approach was used to assess the composition of the ciliate community at a coastal site in the central Adriatic Sea (BioPlan project). Water samples were collected using a Niskin bottle and a 53 µm plankton net. Bulk DNA was extracted by high-throughput sequencing of the nuclear 18S rRNA gene (V4 and V9 regions), in parallel with morphometric identification under a light microscope. The comparative analysis revealed a much better taxonomic resolution of tintinnids by microscopy, suggesting that morphometric identification remains a crucial tool for the detection of ecologically important species in the marine environment. Tintinnid genera such as Rhabdonella, Xystonella, Dadayiella, Favella, and Epiplocylis were confirmed exclusively by microscopic observation. Conversely, metabarcoding proved to be more effective in detecting non-tintinnid ciliates, such as Spirotontonia, Pseudotontonia, Bergeriella ovata (V4), Metaurostylopsis, and Mesodinium rubrum (V9), which were not present in morphological analyses. These discrepancies between approaches likely reflect the limitations of the reference databases, low genetic divergence in the target regions, and potential DNA degradation, particularly in net samples. Our results highlight the limitations of metabarcoding when applied independently and support the use of integrated morpho-molecular protocols in protist biodiversity studies. They also emphasise the importance of expanding curated sequence databases. Understanding the fine taxonomic structure and ecological dynamics of ciliate communities is critical for monitoring ecosystem health and predicting ecological responses to environmental change in marine ecosystems.

The extensive morphological variation and broad distribution of Dendrobium species often hinder accurate identification, leading to potential misapplication and supply challenges in medicinal use. This study aimed to investigate the correlation between phytochemical composition and DNA barcoding markers based on ITS and matK to improve identification in twenty-one medicinal Dendrobium species. Phytochemical data compiled from databases were analyzed through hierarchical clustering to construct a chemical dendrogram. Seven chemical clusters were identified, characterized by key structural compounds such as bisbibenzyls, fluorenones, flavonols, phenanthrenes, and 9,10-dihydrophenanthrenes. Each chemical cluster revealed characteristic profiles of major and minor compounds among Dendrobium species. The ITS-based phylogenetic tree proved more reliable than matK for distinguishing Dendrobium sections. Correlation analysis between the phytochemical dendrogram and the ITS-based phylogenetic tree using the Fowlkes–Mallows Index (FMI) yielded a value of 0.5536, reflecting a noteworthy correlation that highlights the underlying relationship between chemical and genetic clustering. A strong correlation was observed when a phylogenetic clade contained a single chemical cluster. This pattern was evident in clades E and F, which were associated with chemical cluster 7. Cluster 7 comprised eight species, representing the largest group among all clusters. This integrative approach provides a framework for chemical forecasting and supports the authentication and sustainable utilization of Dendrobium resources in traditional medicine.