Identification Of Species Research Articles

Tick-Borne Pathogens in Mongolian Ticks: The High Prevalence of Rickettsia raoultii and Its Public Health Implications.

In Mongolia, a substantial segment of the population is engaged in pastoralism, leading to frequent interactions with livestock and heightened exposure to tick-infested environments. Consequently, ticks and the endemic tick-borne pathogens (TBPs) they harbor present significant health threats. In May 2021, we conducted a study to evaluate the prevalence of 9 TBPs in ticks collected from four regions in Mongolia: Uvurkhangai, Tuv, Bayan-Ulgii, and Khentii. The pathogens surveyed included Anaplasma, Rickettsia, Bartonella, Borrelia, Ehrlichia, Babesia, Toxoplasma, Theileria, and lumpy skin disease virus. Molecular analysis of 1,142 ticks revealed that, although the majority of TBPs were not detected, 89.1% of the ticks were positive for Rickettsia. Genetic characterization using the 16S rRNA and gltA genes identified the pathogen identified the pathogen Rickettsia raoultii. Species identification indicated a predominance of Dermacentor nuttalli (70.0%) and Dermacentor silvarum (30.0%). These findings highlight the extensive prevalence of R. raoultii in Mongolia and underscore the urgent need for heightened awareness and preventive measures due to the high level of interaction between livestock and humans. The genetic similarity of R. raoultii sequences to those found in neighboring countries suggests potential cross-border transmission, underscoring the importance of conducting similar research in surrounding regions. This study advances our understanding of the epidemiology of tick-borne diseases in Mongolia and can inform public health strategies aimed at mitigating the risks associated with these infections.

Contrasting pollution responses of native and non-native fish communities in anthropogenically disturbed estuaries unveiled by eDNA metabarcoding

Understanding the influence of environmental pollutants on the assembly mechanisms of estuarine fish communities is vital for addressing conservation challenges in these biodiverse ecosystems. Although significant research has explored the toxic impacts of pollutants such as petroleum, heavy metals, and eutrophication on individual species and populations, their effects on community assembly processes and the differential responses of native versus non-native fish at the meta-community level remain inadequately understood. This study utilized environmental DNA (eDNA) metabarcoding to analyze fish community diversity across 28 subtropical estuaries in China, assessing how these pollutants affect community composition and assembly mechanisms. Results indicated that eDNA was 2.54 times more effective than traditional methods in species identification, while also enabling the detection of a higher number of non-native fish species and more diverse functional guilds within estuarine ecosystems. A significant distance decay pattern (p < 0.05) was observed among native fish, whereas non-native species exhibited non-significant patterns. Neutral and null models showed that non-native species had significantly higher migration rates (0.005939 vs 0.001757) and a greater contribution of stochastic processes (82.38% vs 70.59%) compared to native species. Additionally, distance-based redundancy analysis (db-RDA), variance partitioning analysis (VPA), and correlation analyses revealed that native species were strongly constrained by environmental factors, particularly oil, Hg, Zn, Pb, Cr6+, and NH4+, while non-native species displayed notable resilience to these pollutants. These findings highlight the potential for non-native species to disproportionately influence community dynamics and assembly through unrestricted random dispersal amid environmental disturbances. This research clarifies the contrasting ecological responses of native and non-native fish communities to anthropogenic pressures in estuarine environments, offering essential insights into ecosystem resilience and informing biodiversity conservation strategies in rapidly changing coastal ecosystems.

Tetracycline degradation for wastewater treatment based on ozone nanobubbles advanced oxidation processes (AOPs) – Focus on nanobubbles formation, degradation kinetics, mechanism and effects of water composition

Presence of pharmaceuticals, especially antibiotics, in industrial and domestic effluents causes serious damage to the environment. Classic wastewater treatment processes, in particular conventional biological treatment methods, are not sufficient to rapidly eliminate antibiotics. Typically, Advanced Oxidation Processes (AOPs) based on activation of hydrogen peroxide, ozone or persulfate for production of particular type of radical species or singlet oxygen are used. A one of cutting-edge technologies to increase effectiveness of AOPs based on ozone are nanobubbles based processes. Thus, this paper focuses on utilization of ozone in the form of nanobubbles for degradation of tetracycline (TC). The effects of several reaction parameters, such as antibiotic concentration, ozone intake, pH, presence of salts, were investigated. This study revealed that the presence of ozone nanobubbles had a substantial positive impact on the degradation of TC. This improvement may be attributed to the enhanced mass transfer and the production of reactive radicals that occur during the collapse of the nanobubbles. Identification of radical species revealed a significant contribution of hydroxyl radicals in the degradation of the antibiotic. AOP process based on O3 nanobubbles was most (•OH) and superoxide (O2–) effective with 100 % degradation efficiency of 100 mg/L TC within 20 min at 8 mg/L concentration of ozone. Based on identified by LC-MS intermediates a degradation mechanism has been described. Degradation of TC and intermediates transformations included methylation, hydroxylation, ring-opening steps as well as cleavage of C-N bonds. This research introduces a novel technique combining nanobubbles with advanced oxidation processes (AOPs), which is anticipated to provide enhanced efficiency and environmental sustainability.

Rapid and efficient removal of organophosphorus pollutant and recovery of valuable elements: A boosted strategy for eliminating organophosphorus from wastewater

Considering the significant hazards of organophosphorus compounds (OPs) and the potential crisis of phosphorus (P) resource shortage, there is a great necessity to develop economically feasible, highly effective, and sustainable strategies to remove OPs and recover P resources. In this study, low-cost microscale zero-valent iron (mZVI) was used to activate hydrogen peroxide for the rapid and efficient elimination of Tetrakis(hydroxymethyl)phosphonium sulfate (THPS) from the aquatic environment. Compared to the conventional Fenton reaction and commercial mZVI, mZVI/H2O2-based Fenton-like reaction exhibited superior removal performance for THPS. The removal mechanism of the mZVI/H2O2 system for THPS was thoroughly elucidated through the identification of reactive oxygen species, characterization analysis, and theoretical calculation. Furthermore, the valuable components of the degradation products were successfully recovered through thermally induced precipitation of the sample followed by high-temperature calcination. The mZVI/H2O2 system has demonstrated significant advantages in removing organic compounds from various types of actual wastewater and improving the biodegradability of the wastewater. This study presented an environmentally friendly and highly efficient strategy to eliminate OPs pollution and recover P resources. It also provided an easy-to-operate method for remediating actual industrial wastewater.

Elemental profile of Ipê species to support timber tracing in Amazon Rainforest

Accurate identification of geographical origin and species of wood is crucial for endorsing laws to combat illegal trade. Here, the multi-element profile of Ipê species of economic interest in Brazil was investigated by neutron activation analysis (NAA). Ba, Br, Ca, Ce, Co, Eu, Fe, K, La, Na, Rb, Sc, Sm, Sr and Zn were determined in the heartwood and sapwood tissues. The Random Forest algorithm identified Sr, Ba, Sm, La, Ce and Br as the most important elements for discriminating species. The analyses revealed significant differences between species and wood tissues, indicating that multi-element profile has great potential as a forensic tool. The establishment of comprehensive reference databases that cover large geographical areas and take into account the natural variability between species is fundamental to achieving this goal.

Novel visual detection technique of isothermal Ligation-dependent probe combined with hybridization chain reaction and hydrogel for the wild edible mushroom component

Multiplex ligation-dependent probe amplification (MLPA) is a high-throughput detection technology with superior specificity and sensitivity, which combines ligation-dependent probe hybridization and PCR amplification. However, the whole process requires repeated temperature change, and the detection results need high resolution agarose electrophoresis or capillary electrophoresis, which limits its application. In viewing of its advantages, an isothermal reaction process based on that combined Hybridization chain reaction (HCR) and hydrogel technology is developed, in order to achieve visual output of detection results without relying on any large instruments and equipment. The edible mushroom is taken as the detection target, and the LP probes were designed according to its reported endogenous reference gene. Through optimizing the reaction conditions, only the samples contained target component, the result showed irregular hydrogel. This visual detection system had good specificity, which detection limit was as low as 0.32 ng/μL. The development of this method provides a new strategy for species identification and adulteration detection of edible mushroom, even for more DNA target detection.

Detection of Theileria in cattle ticks (Rhipicephalus microplus) (Canestrini, 1888) in upper-northeastern Thailand

Theileria parasites were investigated from cattle ticks (Rhipicephalus microplus (Canestrini, 1888)) collected in 12 provinces in upper-northeastern Thailand based on the sequences of 18S rRNA and MPSP gene. Polymerase chain reactions (PCRs) and sequencing specific regions for the 18S rRNA gene revealed two species of Theileria pathogens; T. orientalis (n=42) and T. sinensis (n=31) with 94.50−100% identity. In the provinces of upper-northeastern Thailand, the nucleotide diversity of Theileria's 18S rRNA for T. sinensis and T. orientalis were 0% and 1.3%. respectively. The MPSP gene was used to categorize the T. orientalis genotypes. The sequences were compared with those available in the public database (GenBank) for species identification. Phylogenetic trees of Theileria were constructed from the MPSP gene sequences of our amplicons and those available in GenBank using maximum-likelihood and neighbor-joining analyses. The results revealed three identified genotypes: type 3, 5, and 7. Although the main carriers of T. orientalis are ticks in the genus Haemaphysalis, T. orientalis was the most frequently found in R. microplus in upper-northeastern Thailand. Theileria was frequent in Nong Khai, Mukdahan, and Loei, three Thai provinces that bordered the Lao PDR close to the Mekong River. Epidemiological surveys and control strategies in this region should be considered.

An expert system for species discrimination and grade identification of fish maw

Fish maw, derived from dried swim bladders of fish, is valued for its nutritional and medicinal properties, which has led to an increased market demand. However, price variability based on species and grades has results in unethical practices such as counterfeiting and mislabeling, highlighting the need for reliable quality authentication. To address this, an expert system using MATLAB software has been developed. This system employs nuclear magnetic resonance (NMR) technology and pattern recognition methods to identify fish maw species and classify their grade. The study analyzed ten species across three grades of fish maw, identifying 43 nutritional components from NMR spectra, including sugars, amino acids, fatty acids, organic acids, and vitamins. Univariate statistical analysis was integrated with multivariate statistical analyses, including principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and orthogonal partial least squares (OPLS-DA) using SIMCA software, to develop models for species and grades identification. A four-dimensional volcano map was constructed to highlight characteristic components of various fish maws, resulting in an NMR database for common fish maws. The expert system’s accuracy was validated with new samples, achieving 92.6% for species identification and 90.0% for grade classification. This study provides a valuable tool for the quality evaluation of fish maw and a scientific basis for market regulation.

Mosquito species identification accuracy of early deployed algorithms in IDX, a vector identification tool

Mosquito-borne diseases continue to pose a great threat to global public health systems due to increased insecticide resistance and climate change. Accurate vector identification is crucial for effective control, yet it presents significant challenges. IDX - an automated computer vision-based device capable of capturing mosquito images and outputting mosquito species ID has been deployed globally resulting in algorithms currently capable of identifying 53 mosquito species. In this study, we evaluate deployed performance of the IDX mosquito species identification algorithms using data from partners in the Southeastern United States (SE US) and Papua New Guinea (PNG) in 2023 and 2024 . This preliminary assessment indicates continued improvement of the IDX mosquito species identification algorithms over the study period for individual species as well as average regional accuracy with macro average recall improving from 55.3% [Confidence Interval (CI) 48.9, 61.7] to 80.2% [CI 77.3, 84.9] for SE US, and 84.1% [CI 75.1, 93.1] to 93.6% [CI 91.6, 95.6] for PNG using a CI of 90%. This study underscores the importance of algorithm refinement and dataset expansion covering more species and regions to enhance identification systems thereby reducing the workload for human experts, addressing taxonomic expertise gaps, and improving vector control efforts.

Hidden under darkened wings: the identity of Doryctobracon fluminensis (Lima, 1938) (Hymenoptera: Braconidae) and a new species of the genus from Brazil

Doryctobracon fluminensis (Lima) is one of several species of braconid parasitoids of fruit flies occurring in Brazil. Nearly 60 years passed after the original description by Lima (1938) until the publication of new records of D. fluminensis in several Brazilian states. Species identifications in these records were generally based on the darkened (infumate) fore wings, which has become the main character used in identification keys for recognizing D. fluminensis. After examination of the holotype of D. fluminensis, and comparison with several Brazilian specimens, we concluded that infumate fore wing alone is insufficient for separating species in the genus, as other Doryctobracon species also possess this feature. Here, we describe a new species, Doryctobracon simulatus Marinho sp. nov., based on specimens previously identified as D. fluminensis; redescribe D. fluminensis (Lima); and redefine the limits of D. maculatus Marinho. The misidentification discovered in the only two voucher specimens available indicates that other records of D. fluminensis in Brazil may also be incorrect, although those identifications cannot be verified. Our results demonstrate the importance of preserving voucher specimens in collections for future studies. An updated key to the species of Doryctobracon attacking fruit-infesting tephritids in Brazil is presented.

The genus Toyoides Matsumura (Hemiptera: Auchenorrhyncha: Delphacidae): new combination and lectotype designations

In this study, we revised the planthopper genus Toyoides Matsumura (Hemiptera: Auchenorrhyncha: Delphacidae) to include three species. The genus Epunka Matsumura be synonymized with Toyoides based on habitus and genital characteristics. The lectotype specimens have been designated for both T. albipennis and T. bilineata comb. n. Morphological information on T. bilineata comb. n. is presented, with photographs of the habitus and male genitalia provided. We have also provided the keys for the identification of related genera and species.

Taxonomy and phylogeny of rodents parasitic fleas in southeastern China with description of a new subspecies of Ctenophthalmus breviprojiciens.

Accurate differentiation and identification of flea species are essential for both basic and applied research on fleas, as well as for the diagnosis of flea-borne diseases. However, distinguishing between flea species can be challenging, especially among those with minimal morphological differences. Therefore, some scholars have suggested the necessity of comprehensive revisions to the classification of fleas, incorporating morphological, molecular, and phylogenetic data. In this study, we focused on classifying the rodents' parasitic fleas in southeastern China and provided molecular and phylogenetic data. We also described a new subspecies Ctenophthalmus breviprojiciens fujiansis n. ssp. A total of 392 fleas were collected from 8 species of rodents in 10 counties. Morphologically, they belonged to 10 species, 9 genera and 5 families. Barcode identification based on COI gene and phylogenetic analysis based on five genetic markers (18S rDNA, 28S rDNA, EF-1a, COI, COII) revealed that the molecular and morphological identification of Xenopsylla cheopis, Aviostivalius klossi bispiniformis, Leptopsylla segnis, Monopsyllus anisus and Ctenocephalides felis felis were consistent. The taxonomic status of Neopsylla specialis minpiensis and Peromyscopsylla himalaica sinica as subspecies is questionable due to significant intraspecific genetic distance, and further morphological and molecular data are required to determine if they should be elevated to species level. The molecular identification of C. breviprojiciens n. ssp., N. dispar fukienensis, and Nosopsyllus nicanus could not be completed at this time due to a lack of sequences for related species in existing GenBank databases. Additionally, phylogenetic relationships of 31 species from 9 genera and 5 families of Siphonaptera were inferred based on five molecular markers (18S rDNA, 28S rDNA, EF-1a, COI and COII) using Maximum Likelihood analyses. The analyses revealed that various taxa of Siphonaptera are monophyletic at the subspecies, species, and genus levels. However, at the family level, Leptopsyllidae, Ceratophyllidae, Pulicidae, and Pygiopsyllidae are all monophyletic, while Ctenophthalmidae is paraphyletic. we support the view of some authors that revising the catchall group Ctenophthalmidae and elevating each of its constituent subfamilies to family status.

Highly Specific and Rapid Multiplex Identification of Candida Species Using Digital Microfluidics Integrated with a Semi-Nested Genoarray.

Candida species are the most common cause of fungal infections around the world, associated with superficial and even deep-seated infections. In clinical practice, there is great significance in identifying different Candida species because of their respective characteristics. However, current technologies have difficulty in onsite species identification due to long turnover time, high cost of reagents and instruments, or limited detection performance. We developed a semi-nested recombinase polymerase amplification (RPA) genoarray as well as an integrated system for highly specific identification of four Candida species with a simple design of primers, high detection sensitivity, fast turnover time, and good cost-effectiveness. The system constructed to perform the assay consists of a rapid sample processing module for nucleic acid release from fungal samples in 15 min and a digital microfluidic platform for precise and efficient detection reactions in 35 min. Therefore, our system could automatically identify specific Candida species, with a reagent consumption of only 2.5 μL of the RPA reaction mixture per target and no cross-reaction. Its detection sensitivity for four Candida species achieved 101-102 CFU/mL, which was 10-fold better than conventional RPA and even comparable to a common polymerase chain reaction. Evaluated by using cultured samples and 24 clinical samples, our system shows great applicability to onsite multiplex nucleic acid analysis.

Identification, Pathogenicity, and Fungicide Sensitivity of Colletotrichum Species Associated with Anthracnose on Italian Ryegrass in Southwestern China.

Italian ryegrass (Lolium multiflorum L.) is widely cultivated as an important forage grass worldwide because of its high nutritional value and good palatability. Anthracnose caused by Colletotrichum species was a common and new emerging disease of Italian ryegrass. In this study, 88 Colletotrichum isolates were collected from diseased leaves of Italian ryegrass planting regions in Sichuan, Chongqing, and Guizhou provinces of southwestern China between 2019 and 2022. By the pure culture technique, 15 representative single-spore isolates were obtained for further study. Multilocus phylogenetic analysis coupled with morphological features showed that these isolates were finally identified as six new record species: C. cereale of the C. graminicola species complex, C. fioriniae and C. nymphaeae of the C. acutatum species complex, C. boninense and C. citricola of the C. boninense species complex, and C. nageiae. Pathogenicity tests indicated that all species could induce anthracnose symptoms; of these, C. cereale was more invasive than other species, followed by C. fioriniae, C. nageiae, C. citricola, and C. boninense; C. nymphaeae was the weakest pathogenic species to Italian ryegrass plants (P ≤ 0.05). Fungicide sensitivity assays showed that iprodione, propineb, and oxime·tebuconazole had strong inhibitory effects on the mycelial growth of six Colletotrichum species; in addition, azoxystrobin and fludioxonil also significantly inhibited the mycelial growth of C. nymphaeae and C. fioriniae, respectively. These results provide the basis for the diagnosis and detection in the field, pathogen identification, and management of anthracnose on Italian ryegrass.

GPS telemetry facilitates the identification of forage plant species for endangered Carnaby’s cockatoos (Zanda latirostris) at inland breeding sites and post-breeding dispersal locations in Western Australia

Carnaby’s cockatoos (Zanda latirostris) are an endangered species that has experienced major loss of habitat over the past century. Proponents seeking land clearing approval that may impact Carnaby’s cockatoos need to provide detailed habitat assessment. However, the current forage species list is outdated and generally restricted to the Swan Coastal Plain rather than the Carnaby’s full distribution range. This study provides an updated forage list, including the Swan Coastal Plain and much of the Carnaby’s breeding and non-breeding areas outside this region. Carnaby’s cockatoos were captured and satellite tagged, at five breeding sites in the wheatbelt and Great Southern areas in Western Australia (between 2017 and 2022). Spatial data collected from the tags facilitated the identification of forage plants used by Carnaby’s cockatoos. A total of 44 ‘new’ native plant species were identified as Carnaby’s cockatoo forage species, including five genera that have not previously been recorded. The updated forage list will inform proponents and regulators on the potential use of habitat patches by Carnaby’s cockatoos, aiding the referral process and enabling the protection and conservation of important and diminishing habitat resources.

Enhanced Plant Leaf Classification over a Large Number of Classes Using Machine Learning

In botany and agriculture, classifying leaves is a crucial process that yields vital information for studies on biodiversity, ecological studies, and the identification of plant species. The Cope Leaf Dataset offers a comprehensive collection of leaf images from various plant species, enabling the development and evaluation of advanced classification algorithms. This study presents a robust methodology for classifying leaf images within the Cope Leaf Dataset by enhancing the feature extraction and selection process. Cope Leaf Dataset has 99 classes and 64 features with 1584 records. Features are extracted based on the margin, texture, and shape of the leaves. It is challenging to classify a large number of labels because of class imbalance, feature complexity, overfitting, and label noise. Our approach combines advanced feature selection techniques with robust preprocessing methods, including normalization, imputation, and noise reduction. By systematically integrating these techniques, we aim to reduce dimensionality, eliminate irrelevant or redundant features, and improve data quality. Increasing accuracy in classification, especially when dealing with large datasets and many classes, involves a combination of data preprocessing, model selection, regularization techniques, and fine-tuning. The results indicate that the Multilayer Perception algorithm gives 89.48%, the Naïve Bayes Classifier gives 89.63%, Convolutional Neural Networks has 88.72%, and the Hoeffding Tree algorithm gives 89.92% accuracy for the classification of 99 label plant leaf classification problems.

Sexual Dimorphism in Wing Shape and Its Impact on Conspecific Identification of Neotropical Fannia Species (Diptera: Fanniidae)

Neotropical species of the genus Fannia remain poorly known despite their potential collection as biological evidence in criminal investigations. This is partly due to taxonomic difficulties and a lack of specialists. Identifying neotropical species of Fannia relies mainly on the classical morphological characters of adult males, as females show high similarity between species. Here, landmark-based geometric morphometrics of the wing could constitute an additional tool for associating adult females with conspecifics in this genus. In this study, we used a reference dataset of males belonging to ten putative species of Fannia from Colombia and molecular data to test this hypothesis. We found a strong wing shape sexual dimorphism, resulting in an almost perfect sex assignment based solely on this trait. However, the differences in wing shape between sexes were greater than those between species, making conspecific identification difficult. Our data show that wing shape could only feasibly be used for identifying adult males and females of F. lamosca, and males of F. dorsomaculata and F. pseudoconstricta. Low discrimination scores among remaining species may be partly explained by high intraspecific variation, slight wing shape differences among closely related species, or sampling bias. Although this study provides the first wing size and shape comparison among neotropical Fannia species, more samples and species are needed to validate these findings and identify the potential factors influencing this trait. Furthermore, the wing shape sexual dimorphism across Fannia species suggests different life-history strategies between sexes and possible genetic canalization mechanisms.

An antimicrobial drug recommender system using MALDI-TOF MS and dual-branch neural networks.

Timely and effective use of antimicrobial drugs can improve patient outcomes, as well as help safeguard against resistance development. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is currently routinely used in clinical diagnostics for rapid species identification. Mining additional data from said spectra in the form of antimicrobial resistance (AMR) profiles is, therefore, highly promising. Such AMR profiles could serve as a drop-in solution for drastically improving treatment efficiency, effectiveness, and costs. This study endeavors to develop the first machine learning models capable of predicting AMR profiles for the whole repertoire of species and drugs encountered in clinical microbiology. The resulting models can be interpreted as drug recommender systems for infectious diseases. We find that our dual-branch method delivers considerably higher performance compared to previous approaches. In addition, experiments show that the models can be efficiently fine-tuned to data from other clinical laboratories. MALDI-TOF-based AMR recommender systems can, hence, greatly extend the value of MALDI-TOF MS for clinical diagnostics. All code supporting this study is distributed on PyPI and is packaged at https://github.com/gdewael/maldi-nn.

VeTAPRH: A Taxonomic Assignment Protocol for Vertebrates Applied to eDNA Metabarcoding Data, Including Molecular, Taxonomic and Ecological Criteria

Exploring biodiversity requires meticulous species identification within specific environments. Environmental DNA (eDNA) refers to DNA extracted from environmental samples (Taberlet et al. 2018). Using metabarcoding with eDNA enables detailed taxonomic inventories (Valentini et al. 2009, Haderlé et al. 2024). After sample collection, DNA is extracted, the targeted barcode is amplified, and obtained amplicons are sequenced, producing millions of sequences. Bioinformatics processing is then a particularly important step in the analysis used to clean, structure sequences and complete taxonomic assignments. However, computerized protocols face limitations, including taxonomic misidentifications due to intraspecific variation, such as in the Delphininae (Alfonsi et al. 2013), or incomplete reference databases (e.g. Hleap et al. 2021, Meglécz 2023). Relying on custom databases may help but can miss unexpected taxa range shifts (Gold et al. 2021, Jung et al. 2016). Including ecological data (Coissac et al. 2012, Deiner et al. 2017, Blackman et al. 2023) or integrating multiple databases (Bourret et al. 2023) enhances accuracy, and expert validation remains essential (Clarke et al. 2017, Porter and Hajibabaei 2018). Our study aims to develop a taxonomic assignment protocol for vertebrate Molecular Operational Taxonomic Units (MOTUs)/Amplicon Sequence Variants (ASVs), called VeTAPRH (Vertebrates Taxonomic Assignment Protocol), to verify automated assignments from metabarcoding pipelines. This protocol generates successive species lists, first built on the basis of molecularly similar taxa, then successively refined on the basis of taxonomic data and plausibility of local distribution, compiled from specialized databases. By cross-referencing these criteria, we establish a list of hypothetical taxa for each MOTU, including those absent from reference databases. Initially presented as a flowchart, this protocol will lay the groundwork for a digital tool to support taxonomists.

Decomposition dynamics and Calliphoridae succession in forest and urban environments of Constantine (Northeast of Algeria): implications for forensic entomology

ABSTRACT Forensic entomology serves a vital role in criminal investigations, contributing to the determination of time of colonization (TOC), minimum post-mortem interval (minPMI), and other forensic evidence related to the decomposition process. This study aimed to identify Calliphoridae species attracted to carcasses in two distinct locations in Algeria and to understand their role in forensic investigations. Two sites in Constantine were selected: a forest in Jebel Ouahch and an urban area in Chaabat Erssas. Two dog carcasses, an adult and a puppy, were monitored for decomposition stages. The puppy carcass reached the dry stage faster than the adult carcass, with decomposition rates varying between sites. A total of 594 Calliphoridae flies were collected, including Calliphora vicina, Calliphora vomitoria, Calliphora subalpina, Lucilia sericata, Lucilia illustris, Lucilia silvarum, and Chrysomya albiceps. These species were present in both locations, with Ch. albiceps being most prevalent, especially on the puppy carcass. Initial blowfly colonization occurred within four hours, with L. sericata and Ch. albiceps particularly abundant. This study underscores the importance of accurate species identification and environmental reconstruction in improving the precision of forensic estimates. Continued research on species phenology and the impacts of climate change will further enhance the applicability of forensic entomology in Algeria.