Morphometric Approach Research Articles

Morphometric and taxonomic approach to describe Heterospio variabilis (Annelida, Longosomatidae), a new species with three size-dependent morphotypes, from the Gulf of California, Eastern Pacific.

The Longosomatidae, a poorly known polychaete family, includes only 23 recognized species; in this study, based on morphometric and taxonomic analyses, we describe a new species with three morphotypes: Heterospio variabilis from the Gulf of California, Mexico. The specimens examined exhibit large morphological variations but were clearly separated from close species due to a unique combination of morphological characters: chaetiger 9 as the first elongated chaetiger, four to eight branchial pairs; chaetae from chaetiger 10 forming rings in two rows, posterior row with thin and robust capillaries, anterior row with subuluncini, aristate spines, acicular spines and thick acicular spines. With the discriminant analysis, carried out on 11 morphometric characters, the presence of three morphological groups were recognized (Wilks' lambda= 0.093, p = 0.0001). However, the variables selected to discriminate the specimens (partial Wilks' lambda >0.57) were correlated to their size: number of branchiae, body width, prostomium width, rate length CH9/CH1-CH8, length CH1-CH8 and length CH9 (r>0.5). So, we concluded that they belong to a single species with three morphotypes: morpho A with eight branchial pairs, morpho B with 5-6-7 pairs and morpho C with 4 pairs. No correlations between the distribution of the distinct morphotypes along the eastern gulf shelf and the environmental conditions where they settle were detected.

448 Deformable Medial Modeling to Generate Novel Shape Features of the Placenta Using Automated versus Manual Segmentations

OBJECTIVES/GOALS: In this study, we implemented deformable medial modeling as a morphometric approach in first trimester placentas to characterize morphometric differences between fully automated and manual segmentations. METHODS/STUDY POPULATION: Twenty placentas from singleton pregnancies between 11-14 weeks’ gestation were manually and automatically segmented from 3D ultrasound volumes. Automated segmentations were produced by a trained convolutional neural network pipeline. Dice overlap scores and volumes were computed between manual and automated segmentations. Deformable medial modeling was applied to both manual and automated segmentations to produce the following metrics: maternal and chorionic surface areas (SA), thickness, circumference, and diameter along the generated medial surface. Placental non-planarity was also determined as the greatest medial surface height difference. A paired t-test and simple linear regression was performed between manual and automated segmentations for each shape metric. RESULTS/ANTICIPATED RESULTS: Mean placental volume measurements between manual and automated segmentations were similar, with a percent difference of 3.28% and a mean Dice overlap score of 0.85 ± 0.07. There were strong, statistically significant (p <0.01) linear correlations with chorionic and maternal SA, SA difference, thickness, circumference, medial surface diameter, and medial surface height difference. No significant differences were noted with chorionic SA, thickness, circumference, maximum medial surface diameter, or medial surface height difference. However, statistically significant differences (p <0.01-0.03) were noted in maternal SA, SA difference, and mean medial surface diameter. Despite these differences, mean percent difference for all morphometric parameters was less than or equal to 10%. DISCUSSION/SIGNIFICANCE: A deformable medial model evaluate unique global and regional shape placental features with highly correlated values between manual versus automated placental segmentations. However, clinical studies are needed to determine if minor differences would impact the clinical utility of these features as potential indicators of placental function.

Geometric morphometrics casts light on phylogenetic relevance of cephalopod beak morphological.

The feeding organ of cephalopod species, the beak, can be used to reveal important ecological information. In this study, geometric morphometric approaches were employed to investigate the phylogenetic relevance and classification effect of beak lateral profile shape. The two-dimensional beak morphologies of 1164 pairs of 24 species from 13 genera and five families were constructed, and their evolutionary relationships and taxonomic status were confirmed using geometric morphometrics and molecular biology approaches. We also assessed the phylogenetic signals of beak shape. The analysis results show shape variation in the beak mainly in the rostrum, hood, and lateral wall. The overall shape parameters (all PCs) of the upper and lower beak are more useful for species identification. The shapes of the upper and lower beak show a strong phylogenetic signal, and the phenogram based on the beak shape basically reflected the families' taxonomic positions. We also hypothesized that the shape variation in the beaks of cephalopods may be ascribed to genetic and environmental differences. In summary, beaks are a reliable material for the classification of cephalopod species. Geometric morphometric approaches are a powerful tool to reveal the identification, phylogenetic relevance and phenotypic diversity of beak shape in cephalopods.

A Geometric Morphometrics Approach to the Study of Natural Variations and Hybrid Detection in Populations of Alnus incana (L.) Moench and Alnus rohlenae Vít, Douda and Mandák.

Landmark-based geometric morphometrics (GM) was used to examine, for the first time, spontaneous hybridization between Alnus incana (L.) Moench and Alnus rohlenae Vít, Douda and Mandák, and to assess inter- and intrapopulation variability in leaf shape, leaf size and venation in natural populations in Serbia (Western Balkans). Two geographically distant (30 km) and two close (1.2 km) populations were selected to examine hybridization. The variability in leaf shapes was assessed by canonical variate analysis and linear discriminant analysis performed on the symmetric component of variation. Covariation between the symmetric component of shape variation and the number of pairs of secondary leaf veins was investigated with partial least squares analysis. Static allometry was examined for the first time in the genus Alnus Mill. A higher proportion of A. incana leaves was classified as A. rohlenae in geographically close populations, which is in accordance with the hypothesis about spontaneous hybridization. No single leaf of A. rohlenae was classified as A. incana, indicating that putative hybrids can only be found in grey alder populations. This study demonstrates that GM is a powerful tool for species delimitation and hybrid detection in the genus Alnus and it can be used for preliminary screening in hybrid zones.

A Morphometric and Multivariate Analysis Approach to Prioritization of Sub-Watershed: A case study on Muzaffarpur District of Bihar, India

Aim of the studyPrioritization of sub-watershed of Muzaffarpur District of Bihar (India) using Multivariate AnalysisMaterial and methodsPCA and WSA statistical tools apply to find out the important parameters to Prioritization of sub-watershedResults and conclusionsPCA, WSA integration results in more dynamic, effective, and efficient solutions. The Muzaffarpur District's decision-makers can use this useful knowledge to establish management methods that will lessen and prevent land degradation.

Flood hazard mapping using a GIS-based morphometric analysis approach in arid regions, a case study in the Red Sea Region, Egypt

Flash floods are a major threat to life and properties in arid regions. In recent decades, Egypt has experienced severe flash floods that have caused significant damage across the country, including the Red Sea region. The aim of this study is to map the flood hazards in flood-prone areas along the Red Sea region using a Geographic Information System (GIS)-based morphometric analysis approach. To evaluate the flood hazard degree, the adopted methodology considers various morphometric parameters such as basin area, slope, sinuosity index, shape factor, drainage intensity, circularity ratio, and curve number. GIS techniques were employed to delineate the watershed and the drainage network. The delineated watershed was used together with the digitized maps of soil and land use types to estimate the curve number and the morphometric parameters for each subbasin. The flood hazard degrees are calculated based on the considered morphometric parameters and distinguished based on a five-degree scale ranging from very low to very high. Results indicate that 47% of the study area has a very high flood hazard degree. Furthermore, morphometric analysis results align with the runoff results simulated by a hydrological model, where, for example, basins with a high to very high hazard degree exhibited high runoff. This suggests the influence of physical characteristics on the hydrological behavior of the watershed and further validates the morphometric analysis presented in this work. The results presented here can help policy planners and decision-makers develop appropriate measures to mitigate flash floods and achieve sustainable development in arid regions.

Considering Decoupled Phenotypic Diversification Between Ontogenetic Phases in Macroevolution: An Example Using Triggerfishes (Balistidae).

Across the Tree of Life, most studies of phenotypic disparity and diversification have been restricted to adult organisms. However, many lineages have distinct ontogenetic phases that differ from their adult forms in morphology and ecology. Focusing disproportionately on the evolution of adult forms unnecessarily hinders our understanding of the pressures shaping evolution over time. Non-adult disparity patterns are particularly important to consider for coastal ray-finned fishes, which can have juvenile phases with distinct phenotypes. These juvenile forms are often associated with sheltered nursery environments, with phenotypic shifts between adults and juvenile stages that are readily apparent in locomotor morphology. Whether this ontogenetic variation in locomotor morphology reflects a decoupling of diversification dynamics between life stages remains unknown. Here we investigate the evolutionary dynamics of locomotor morphology between adult and juvenile triggerfishes. We integrate a time-calibrated phylogenetic framework with geometric morphometric approaches and measurement data of fin aspect ratio and incidence, and reveal a mismatch between morphospace occupancy, the evolution of morphological disparity, and the tempo of trait evolution between life stages. Collectively, our results illuminate how the heterogeneity of morpho-functional adaptations can decouple the mode and tempo of morphological diversification between ontogenetic stages.

Implementation of geometric morphometry in the study of shapes of Bronze Age sickles from the Volga-Ural region

This article aims at evaluating the potential of geometric morphometry by means of an example of analysis of shapes of the Bronze Age sickles from the Volga-Ural region, as compared with the traditional morphometric approach. For the study, cast bronze sickles with hooks, categorized by V.A. Dergachev and V.S. Bochkarev into the Ibrakaevo, Derbeden, Perelyub and Yavlenka types using the traditional morphometric approach, have been selected. The analysis was applied to only full drawings of the items, including reconstructed ones, while fragmented items were not considered. The sample constitutes 167 objects: 86 Ibrakaevo, 49 Derbeden, 24 Perelyub, and 8 of the Yavlenka type. Application of geometric morphometry tools shows that, within the sample, three main forms can be easily identified, with the exception of the sickles earlier attributed to the Yavlenka type, probably due to their small number. Prepara-tion of primary files for recording landmark coordinates and processing of files with recorded coordinates were carried out in the tpsUtil program. The analysis of characteristics of changes of the forms was carried out using the principal component method in the MorphoJ program. Summarizing the obtained results, we can say that the principal components method has been able to identify three main variations of the objects. Most clear are the differences between the Ibrakaevo and Derbeden types, which show virtually no overlap. An intermediate position between them is taken by the Perelyub type, which is also distinctively grouped in the graph being close to the Ibrakaev group. In terms of identifying individual types, the results of the study are rather consistent with the data obtained by the tradi-tional morphometry. At the same time, it is possible to trace the vectors of shape variability for all types of tools based on three main components. Overall, it can be concluded that the method of geometric morphometry demonstrates its efficiency for the analysis of shapes of metal sickles and in future it could be applied to wider sample groups.

Spatial Modelling of Urban Wind Characteristics: Review of Contributions to Sustainable Urban Development

Wind, a renewable resource with growing importance in the contemporary world, is considered a capable tool for addressing some of the problems linked with rapid urbanization, unsustainable development, and climate change. As such, understanding modelling approaches to wind characteristics in cities becomes crucial. While prior reviews delve into the advancements in reduced-scale models and computational fluid dynamics simulations, there is scant literature evaluating large-scale spatial modelling of urban wind environments. This paper aims to consolidate the understanding of spatial modelling approaches to wind characteristics in cities by conducting a systematic literature review with the PRISMA protocol to capture the contributions to sustainable urban development. The reviewed articles are categorized under two distinctive approaches: (a) studies adopting the wind morphometric approach, encompassing theoretical foundations, input factors, and computation methods and (b) studies adopting the urban climate mapping approach, centering on the amalgamation of wind with urban microclimate analysis. The findings suggest that wind morphometric methodologies hold considerable promise due to their straightforward calculations and interpretability. Nonetheless, issues related to data precision and accuracy challenge the validity of these models. This review also probes into the implications of these two distinctive approaches for urban planning and policymaking, advocating for more sustainable urban development.

Quantifying hominin morphological diversity at the end of the middle Pleistocene: Implications for the origin of Homo sapiens.

The Middle Pleistocene (MP) saw the emergence of new species of hominins: Homo sapiens in Africa, H. neanderthalensis, and possibly Denisovans in Eurasia, whose most recent common ancestor is thought to have lived in Africa around 600 ka ago. However, hominin remains from this period present a wide range of morphological variation making it difficult to securely determine their taxonomic attribution and their phylogenetic position within the Homo genus. This study proposes to reconsider the phenetic relationships between MP hominin fossils in order to clarify evolutionary trends and contacts between the populations they represent. We used a Geometric Morphometrics approach to quantify the morphological variation of the calvarium of controversial MP specimens from Africa and Eurasia by using a comparative sample that can be divided into 5 groups: H. ergaster, H. erectus, H. neanderthalensis, and H. sapiens, as well as individuals from current modern human populations. We performed a Generalized Procrustes Analysis, a Principal Component Analysis, and Multinomial Principal Component Logistic Regressions to determine the phenetic affinities of the controversial Middle Pleistocene specimens with the other groups. MP African and Eurasian specimens represent several populations, some of which show strong affinities with H. neanderthalensis in Europe or H. sapiens in Africa, others presenting multiple affinities. These MP populations might have contributed to the emergence of these two species in different proportions. This study proposes a new framework for the human evolutionary history during the MP.

A geometric morphometric approach to distinguish ferret from polecat and its application to an archaeological specimen from Mechelen (Belgium)

The inability to differentiate skeletal remains belonging to the ferret from those of its wild ancestor, the European polecat, presents a particular challenge for zooarchaeologists which currently hinders a better understanding of ferret domestication history. Using a geometric morphometric approach on the mandible, this study provides a new method to distinguish the two forms. Despite a small sample size and some overlap in the dataset, this method allowed the identification of a (post)medieval specimen from Mechelen (Belgium) as a wild polecat. Results demonstrate that ferrets can largely be distinguished from polecats based on mandibular morphology.

Size at Sexual Maturity of Deep-Sea Unexploited Caribbean Metanephrops binghami (Boone, 1927) and Overexploited Mediterranean Nephrops norvegicus (Linnaeus, 1758) Using Morphometric and Gonadal Staging Approaches

The deep-sea Caribbean lobster (Metanephrops binghami) and the Norway lobster (Nephrops norvegicus) are Nephropidae species of high commercial interest. Although the first one still remains unexploited, the second is overexploited in the Mediterranean Sea. For effective fisheries management, size at sexual maturity is an essential indicator to protect immature individuals from exploitation. The estimation of this indicator can, however, be biased due to the difficulty of differentiating juveniles from adults by their size structure due to the natural process of molting. This study aims to estimate the size at sexual maturity of M. binghami and N. norvegicus females by comparing the effectiveness of the morphometric method versus the macroscopic evaluation of gonad maturity. Samples of M. binghami were collected from the Colombian Caribbean Sea in August and December 2009, March and May 2010, and August 2020 to May 2021. Samples of N. norvegicus were collected from the northwestern Mediterranean Sea from 2019 to 2022. Similar sizes at sexual maturity were found for M. binghami between the morphometric approach (ranging from 28.6 to 33.9 mm cephalothorax length, CL) and the gonadal staging approach (31.4 mm CL). Conversely, for N. norvegicus, the morphometric approach yielded higher measurements (between 27.2 and 30.4 mm CL) than the gonadal approach (26.0 mm CL). This discrepancy might stem from the intense fishing overexploitation conditions of N. norvergicus, leading to a physiological adaptation that enables earlier gonadal maturation at faster rates than morphometric adaptation. Further research is required to elucidate these discrepancies and the effect of overexploitation on physiological (i.e., mature gonads) and functional maturity (i.e., capacity to brood eggs at a larger size).

Insular evolution from an archaeological perspective: a case study of Orkney house mouse

Abstract Archaeological material adds a temporal dimension to evolutionary studies that is valuable for elucidating long-term population stability and evolutionary shifts for species closely associated with humans. Here, a two-dimensional geometric morphometrics approach on first upper molars was applied to modern and archaeological samples to assess the evolution of house mice in Orkney, an archipelago north of Scotland. Modern populations included localities in Orkney, north Scotland, and France. Two archaeological sites in Orkney represented the Norse period: Birsay Beachview (Mainland) and Tuquoy (Westray) (10th–14th/15th centuries AD). The archaeological specimens were larger than modern specimens from similar settings, suggesting processes leading to a recent decline in the size of Orkney house mice. Molar morphology and associated non-metric traits distinct to the Orkney lineage were already established in the Norse period, as indicated by morphological similarity of the Birsay samples to modern Orkney and north Scotland populations. Stability of human settlement is likely to influence morphological evolution in house mice. The Birsay site, located in Birsay village, which has been inhabited since Norse times, might represent the ancestral house mouse population in Orkney. Tuquoy, a settlement abandoned by the end of the medieval period, provided samples different from modern house mouse populations in Westray and neighbouring isles.

Morphometric analysis for determination of larval instars in Dermestes frischii Kugelann and Dermestes undulatus Brahm (Coleoptera: Dermestidae).

Dermestes frischii Kugelann, 1792 and Dermestes undulatus Brahm, 1790 are the most abundant species worldwide at outdoor or indoor crime scenes during the dry and skeletal stages of decomposition. The attribution of larval age in these beetles is problematic due to the variable number of instars, which is influenced by environmental factors. In this study, a morphometric approach was used to look for potential morphological features as evidence of larval stages. Breeding and monitoring were performed for both species in an incubator with a preset temperature of 28°C ± 0.5 without a photoperiod. Morphometric measurements were made on 10 larvae per instar for each species using length, width, and thickness parameters. Linear discriminant analysis was then used to generate decision boundaries that clearly separated larval stages. The cross-validation procedure demonstrated that the morphometric approach successfully discriminated adjacent larval stages in both species with high values of sensitivity and specificity. This less-invasive approach could improve the ability to estimate minPMI in forensic studies of Dermestidae beetles. Future studies may extend this approach to other species and establish good practices for collecting and storing specimens for morphometric analysis.

Geometry of posterior larval spiracles to identify medically and forensically important calliphorids in Thailand

Fly identification is the primary step of analysis in forensic entomology. Although morphology and molecular techniques are considered satisfactory methods, some constraints may arise from a financial or even human point of view. Over the past decade, the geometric morphometric approach has been increasingly advocated for the classification and identification of arthropods. This study explored the method for species identification of 800 third-instar larvae of eight blow fly species of medical and forensic importance: Chrysomya chani Kurahashi, Chrysomya megacephala (Fabricius), Chrysomya (Ceylonomyia) nigripes Aubertin, Chrysomya pinguis (Walker), Chrysomya (Achoetandrus) rufifacies (Macquart), Hemipyrellia ligurriens (Wiedemann), Lucilia cuprina (Wiedemann), and Lucilia porphyrina (Walker). Based on the posterior spiracles geometry, the cross-validation revealed a relatively high percentage of correct classification in most species, ranking from 86% to 100%. The results of this study confirmed that the geometric morphometric (GM) analysis of posterior spiracles might be utilized as a larva identification tool. Therefore, this GM method represents one way of overcoming difficulties with the identification of blow fly larvae and can support further studies of these flies.

Ctenocephalides orientis and Ctenocephalides felis in Thailand: Head geometry by species, sex and geography.

Fleas in the genus Ctenocephalides serve as biological vectors or intermediate hosts of microorganisms such as bacteria, rickettsia, protozoa and helminths. Ctenocephalides felis has a worldwide distribution, while C. orientis has long been considered as a subspecies of C. felis in Asia. To help the morphological recognition of these two species and further explore their differences, we used the geometric morphometric approach applied to the head. Both sexes were examined. Five anatomical landmarks of the head were used, and to capture the curvature of the front head, 10 semilandmarks were added. There was a consistent difference in species classification accuracy when considering landmarks only versus their combination with semilandmarks, suggesting the importance of the curve of the head as a taxonomic signal. Using or not the labels in the reclassification analyses, the head shape allowed by itself almost perfect recognition of the two species, in both sexes, even after adjustment for prior probabilities. The same approach disclosed a high level of sexual size and shape dimorphism in both species. The contribution of size variation to the discrimination by shape was much more important between sexes (from 27% to 45%) than between species (from 0.7% to 7.1%). Nevertheless, in our data, size never could represent a way to reliably recognise the sex of an individual, even less its species. Geographical variation in head shape could only be explored for the C. orientis sample. No significant correlation of morphometric variation with geography could be detected, which would be consistent with gene flow between Thai provinces. The geometric morphometric approach of the flea head, when it incorporates head curves, is a promising tool for rapid, economical, and accurate species and sex identification. It is, therefore, a useful tool for future epidemiological and demographic studies.

Computational Fractal-Based Analysis of Brain Tumor Microvascular Networks.

Brain parenchyma microvasculature is set in disarray in the presence of tumors, and malignant brain tumors are among the most vascularized neoplasms in humans. As microvessels can be easily identified in histologic specimens, quantification of microvascularity can be used alone or in combination with other histological features to increase the understanding of the dynamic behavior, diagnosis, and prognosis of brain tumors. Different brain tumors, and even subtypes of the same tumor, show specific microvascular patterns, as a kind of "microvascular fingerprint," which is particular to each histotype. Reliable morphometric parameters are required for the qualitative and quantitative characterization of the neoplastic angioarchitecture, although the lack of standardization of a technique able to quantify the microvascular patterns in an objective way has limited the "morphometric approach" in neuro-oncology.In this chapter, we focus on the importance of computational-based morphometrics, for the objective description of tumoral microvascular fingerprinting. By also introducing the concept of "angio-space," which is the tumoral space occupied by the microvessels, we here present fractal analysis as the most reliable computational tool able to offer objective parameters for the description of the microvascular networks.The spectrum of different angioarchitectural configurations can be quantified by means of Euclidean and fractal-based parameters in a multiparametric analysis, aimed to offer surrogate biomarkers of cancer. Such parameters are here described from the methodological point of view (i.e., feature extraction) as well as from the clinical perspective (i.e., relation to underlying physiology), in order to offer new computational parameters to the clinicians with the final goal of improving diagnostic and prognostic power of patients affected by brain tumors.

Solving taxonomic species complexes of Stevia (Eupatorieae, Asteraceae) in southern central Andes: a morphometric and statistical approach

The South American central Andes have a great richness of Asteraceae, where the genus Stevia (Eupatorieae, Piqueriinae) is also especially diverse. The taxonomy of Stevia species in the southern part of the central Andes pose a challenge because of the unclear delimitation among the species and overlap of morphological characters. Two species complexes were identified for the region comprising north Argentina and southern Bolivia. In this study, our objectives were to employ morphometric analyses to assess the delimitation of taxa within the Stevia alpina Griseb. and Stevia chamaedrys Griseb. complexes. We aimed to identify informative morphological characters and gain insight into the identity of species in each group. To accomplish this, we utilised cluster analysis, principal-component analysis, non-metric multidimensional scaling and univariate analyses to evaluate 37 morphological traits. In total, 200 specimens were examined, covering the entire geographic distribution of each species complex. We found support for the recognition of nine species: Stevia alpina, S. breviaristata Hook. & Arn., S. chamaedrys, S. mandonii Sch.Bip., S. mercedensis Hieron., S. minor Griseb., S. potrerensis Hieron., S. procumbens Hieron. and S. vaga Griseb. A new combination and status is proposed for S. tarijensis Hieron., incorporating it as a variety of S. mandonii, and S. centinelae Cabrera and S. crassicephala Cabrera are merged as synonyms under the name S. alpina. Two neotypes are designated for the names S. mercedensis and S. potrerensis.

Morphometric analysis of spiny lobster (Panulirus penicillatus) and bamboo lobster (Panulirus versicolor) in Sabang Waters

This research aims to analyze morphometrics of (P. penicillatus) and (P. versicolor) in waters of Sabang. A total of 50 samples from each species of P. penicillatus and P. versicolor were used for morphometric analysis using Traditional Morphometric and Truss Morphometric approaches. Morphological analysis indicated that both lobster species are categorized into two distinct groups. Traditional morphometric character testing using ANOVA showed that species differences significantly influence the variation in morphological characters (P<0.05) in both lobsters. Further Duncan post hoc tests revealed that four characters (HL), (CHW), and (CHL)) differed significantly between the species. P. penicillatus shared three characters similar to P. versicolor, namely (HL), (CHW), and (CHL). This similarity is based on the shape of HL, CHW, and CHL in both lobsters. In the Truss Morphometric character testing with ANOVA, it was found that species differences affect the characters significantly (P<0.05). The Duncan post hoc test revealed that five characters ([A1, A3], [B1, B3], [C5, C6], D4, [D5, D6]) differed significantly between species. DFA resulted in 1 function, with an eigenvalue of 27.595, indicating that it explains 100% of the total variance. The morphological characters that significantly contribute to the differentiation between species are A1, B1, C5, A5, and D5.

A geometric morphometric approach to identify uncomplete snake vertebrae from raptor bird feeding remains

The Hungarian meadow viper (Vipera ursinii rakosiensis) is an endangered subspecies of Vipera ursinii, which faces high predation pressure, partially due to avian species. To create a systematic method for estimating the measure of predation pressure, we developed a geometric morphometric approach to identify both undamaged and damaged vertebrae of snake species found in Hungarian meadow viper habitats from raptor feeding remains. We used linear discriminant analysis with a reference material of vertebrae from identified snake species as training data. We also tested its efficiency by predicting the identification results of different simulation levels based on vertebra completeness. We practiced this method on vertebrae of unknown species of snakes obtained from nests and pellets of short-toed snake eagles (Circaetus gallicus, n = 9), common buzzards (Buteo buteo, n = 14) and Montagu's harriers (Circus pygargus, n = 3). The identification approach showed high accuracy, even in the case of missing landmarks to some extent. We identified vertebrae remnants of Natrix natrix (n = 172, 83.9%), Coronella austriaca (n = 10, 4.9%) and V. u. rakosiensis (n = 23, 11.2%). Both, the reptile specialist C. gallicus and the generalist B. buteo proved to be preying on V. u. rakosiensis, while samples of C. pygargus did not contain any snake remains despite of previous observations of V. ursinii predations. Our approach is applicable for other studies and taxa as well, therefore can be a practical tool for classification of incomplete vertebrae, which is otherwise hardly identifiable. Furthermore, it could be applied to help estimate predation pressure on endangered snake species.