Providing clear and detailed morphological descriptions of endemic species in limited areas enables new knowledge of their biology and ecology to be obtained through citizen science. This information can be further used for their protection. Our study presents the first morphological description of the larvae of all three instars of Heterotemna tenuicornis (Brullé, 1836), an endemic species of the Canary Islands that, together with H. britoi García & Pérez, 1996 and H. figurata (Brullé, 1839), belongs to the peculiar genus Heterotemna Wollaston, 1864. Furthermore, we present the first sequences of two mitochondrial genes (COI, 16S) obtained from larval specimens, and cross reference them with sequences from an adult specimen. Phylogenetic analysis of molecular data placed the genus Heterotemna within the genus Silpha Linnaeus, 1758, suggesting paraphyly of Silpha. In our study, we underline the importance of using a combination of morphological description and molecular data, that can be used for barcoding developmental stages which could not otherwise be definitely associated.

La infección por Leucocytozoon no influye en la supervivencia de los polluelos de mochuelo boreal Aegolius funereus. Se ha observado que la infección por Leucocytozoon afecta la ecología y fisiología reproductiva de las aves hospedadoras, pero su influencia en la supervivencia de los polluelos aún no está completamente determinada. Se investigó el efecto de la intensidad de la infección por Leucocytozoon, determinada mediante análisis de muestras de PCR por triplicado, sobre la supervivencia de 256 polluelos de mochuelo boreal (Aegolius funereus) durante un estudio de ocho años. Contrariamente a nuestras expectativas, la probabilidad de supervivencia de los polluelos de mochuelo boreal no se vio influenciada por la intensidad de la infección por Leucocytozoon. La edad de los polluelos y la intensidad de la infección por Leucocytozoon en los padres machos y hembras tampoco afectaron la supervivencia de los polluelos. En cambio, la abundancia de alimento y el orden de eclosión fueron los principales factores que influyeron en la supervivencia de los polluelos. Además, se observó una intensidad de infección por Leucocytozoon significativamente mayor en los padres machos en comparación con las hembras y los polluelos. Se sugiere que una clara división de los roles parentales puede llevar a que las hembras y los polluelos que permanecen dentro de las cajas nido (cavidades) experimenten una menor exposición a vectores potenciales que transmitan parásitos sanguíneos en comparación con los individuos adultos masculinos. Nuestro estudio muestra que la enfermedad de Leucocytozoon puede no ser letal para los polluelos de mochuelo boreal, ya que exhiben una intensidad de infección por debajo del promedio en comparación con sus padres machos.

The worldwide distributed subfamily of rove beetles Silphinae contains two well‐established tribes, based on both morphological and molecular data. The relationships within the tribe Nicrophorini have been mostly resolved; however, the tribe Silphini still lacks a robust phylogeny. Thus, here we resolved the phylogeny of the tribe based on 42 species of the 114 known species, using five molecular markers. Heterotemna tenuicornis Brullé clustered as sister to Silpha tristis Illiger, making the subgenus Silpha Linnaeus paraphyletic. Consequently, Heterotemna Wollaston is considered a junior subjective synonym of Silpha Linnaeus; requiring the following combinations: Silpha ( Silpha ) britoi (García & Pérez), comb. nov. , Silpha ( Silpha ) figurata Brullé, comb. rest., and Silpha ( Silpha ) tenuicornis Brullé, comb. rest. Our estimate of the phylogeny agrees with current generic limits except it revealed that the genus Aclypea Reitter arose from within the genus Silpha , thus making the latter paraphyletic. Some ambiguity remains regarding the confidence of this finding; therefore, we refrain from synonymizing Aclypea until further study. Furthermore, it includes biogeographical information for each genus, which estimates the history of distributions of the Silphini across the Australian, Neotropical, and Oriental regions.

The study aimed to analyze the entomological material collected during 13 autopsies performed on the unidentified cadavers revealed at different stages of decay in the Upper Silesia Region (Poland) over 2016–2022. During the preparation of human tissues for genetic identification, we revealed larvae, puparia, and adult insects in previously undescribed locations: costal cartilage, femur nutrient canals (foramen nutrients), and tooth cavities. The taxonomical assessment was done using morphological examination or DNA barcoding, where necessary. Based on our observations, we conclude that the apical constriction, foramen, and cavities may serve as migration paths inside teeth, and the femur nutrient canals to the bone marrow. The study also revealed that Necrobia ruficollis (Fabricius, 1775) and Pyralidae Latreille, 1802 (Phycitinae) moths can form pupal chambers inside the costal cartilage, indicating that these insects can complete their life cycle inside this cache. We believe that the newly reported locations of carrion insects in human remains may be relevant to forensic entomology, as they provide new opportunities to collect insect evidence.

The Eurasian lynx (Lynx lynx) is listed in CITES Appendix II and is protected under the Bern Convention and the EU Habitats Directive, yet it remains a frequent target of wildlife crime, highlighting the urgent need for reliable identification methods. This study focuses on determination and DNA quantification of the Lynx spp. using quantitative real-time PCR (qPCR). The Llynx Qplex quantification multiplex system effectively distinguishes Lynx spp. from other Feliformia species by targeting mitochondrial and nuclear markers. Additionally, we present the results of the developmental validation of the Llyn STRplex system for individual identification and databasing using six STR loci. This study followed ISFG recommendations for non-human DNA testing and developmental validation guidelines. Both systems demonstrate high sensitivity (5 pg genomic DNA for Llynx Qplex and 30 pg of mtDNA for Llyn STRplex) and high specificity to Lynx spp., confirmed by testing against 16 related Feliformia species. Robustness was evaluated, showing sensitivity to temperature variation, and both repeatability and reproducibility were successfully tested across replicates and conditions. Given that forensic casework often involves degraded and limited biological material, molecular tools must be both sensitive and specific to ensure accurate results. Developing precise and efficient tools is essential for supporting investigations of wildlife crime involving the Eurasian lynx, as well as efforts aimed at conserving the species.

Background/Objectives The leopard (Panthera pardus), an apex predator listed in CITES Appendix I and classified as Vulnerable by the IUCN, is undergoing severe population declines driven by habitat loss, human–wildlife conflict, and illegal trade. Rapid and reliable species and individual identification is critical for conservation and forensic applications, particularly when analyzing highly processed or degraded seized wildlife products, where morphological identification is often impossible. We aimed to develop and validate a robust multiplex quantitative real-time PCR (qPCR) assay combined with a short tandem repeat (STR) system for the species-specific detection and individual identification of P. pardus. Methods The qPCR assay (Ppar Qplex) was designed to target a mitochondrial Cytochrome b (Cyt b) fragment for species confirmation, a nuclear marker (PLP) for general Feliformia detection and quantification, and an artificial internal positive control (IPC) to monitor PCR inhibition. The assay’s performance was validated for robustness, specificity, sensitivity, repeatability, and reproducibility, utilizing DNA extracted from 30 P. pardus individuals (hair and feces) and tested against 18 related Feliformia species and two outgroups. Individual identification was achieved using a set of 18 STR loci and a sex determination system adapted from previously published Panthera panels. Results Validation demonstrated high specificity for the Ppar Qplex: mitochondrial amplification occurred exclusively in P. pardus samples. The nuclear marker consistently amplified across all 18 tested Feliformia species but not the outgroups. The assay showed high analytical sensitivity, successfully detecting DNA at concentrations as low as 1 pg/µL, with consistent results confirmed across different sample types, replicates, and independent users. Furthermore, the STR multiplex successfully generated 30 unique individual profiles using the 18 polymorphic loci and the sex determination system. Conclusions The combined qPCR assay and STR system provide a fast, sensitive, and highly specific molecular framework for rapid leopard detection, quantification, and individual identification from a wide range of sample types. These tools strengthen forensic capacity to combat wildlife crime and provide critical data to support evidence-based conservation management of P. pardus. P. pardus, an apex predator listed in CITES Appendix I and classified as Vulnerable by the IUCN, is undergoing severe population declines driven by habitat loss, human–wildlife conflict, and illegal trade. Rapid and reliable identification of seized specimens is therefore critical for conservation and forensic applications, mainly when products are highly processed. We developed and validated a multiplex quantitative real-time PCR (qPCR) assay targeting the mitochondrial gene Cytochrome b (Cyt b) for species-specific detection. The assay was tested on verified leopard individuals and validated across 18 Feliformia and two outgroup species (Homo sapiens, Canis lupus familiaris). Analytical performance was assessed through robustness, specificity, sensitivity, repeatability, and reproducibility. Mitochondrial amplification occurred exclusively in leopard samples, while nuclear markers amplified consistently across Feliformia but not in outgroup species. The assay’s limit of DNA detection is 1 pg/µL and produces consistent results across replicates, tested types of samples (hair, feces), and independent users, with internal controls confirming the absence of inhibition. In addition, we present the results of successful individual identification using the set of 18 STR loci and the sex determination system. The developed qPCR and STR systems provide a fast, sensitive, and specific solution for leopard detection and quantification, reinforcing forensic efforts against wildlife crime and supporting conservation of P. pardus.