Phylogenetic Information Research Articles

The Elevational Distribution Patterns of Plant Diversity and Phylogenetic Structure Vary Geographically Across Eight Subtropical Mountains.

Mountains have been recognized as biodiversity hotspots and possess strong elevational gradients. Whether these gradients exhibit similar characteristics in the multidimensional diversity patterns across different mountain ranges is a subject of inquiry. Exploring the elevational patterns of the diversity and phylogenetic information of plant species in the different subtropical mountains is necessary. Here, we compiled the elevational patterns of plant diversity occurring in the eight subtropical mountains of China and focused on the assessment of the patterns and determinants of the multi-dimensional diversity and phylogenetic structure in different mountains. We also detected the elevational patterns and their relationship between different groups. The results indicate two main patterns of multi-dimensional diversity: monotonic decrease and hump-shaped, along with the area effect on the species diversity. There is a strong positive link between the non-native and native species of species richness, and significant differences in phylogenetic structure's elevational distribution. We did not find the same rule in the mountains that the plant species in the lowlands indicate phylogenetic overdispersion, and the species in the higher elevation regions indicate phylogenetic clustering. We found that the plants' diversity peak is related to the mountains size, and this result showed that we should pay more attention to the conservation of plant communities in the higher elevation regions for the higher mountains. This study suggested that we should take different protective measures for the subtropical mountains: the lowland regions should be paid more attention in Mt. Lushan (LS), Mt. Guanshan (GS), Mt. Huanggang (HG), and Mt. Fanjing (FJ), and the middle-altitude regions should be of concern for other mountains. This study helps to better understand the elevational gradients of species diversity on different scales and provides supporting scientific basis for biodiversity conservation in the subtropical mountain regions.

Constructing phylogenetic trees for microbiome data analysis: A mini-review

As next-generation sequencing technologies advance rapidly and the cost of metagenomic sequencing continues to decrease, researchers now face an unprecedented volume of microbiome data. This surge has stimulated the development of scalable microbiome data analysis methods and necessitated the incorporation of phylogenetic information into microbiome analysis for improved accuracy. Tools for constructing phylogenetic trees from 16S rRNA sequencing data are well-established, as the highly conserved regions of the 16S gene are limited, simplifying the identification of marker genes. In contrast, metagenomic and whole genome shotgun (WGS) sequencing involve sequencing from random fragments of the entire gene, making identification of consistent marker genes challenging owing to the vast diversity of genomic regions, resulting in a scarcity of robust tools for constructing phylogenetic trees. Although bacterial sequence tree construction tools exist for upstream bioinformatics, many downstream researchers—those integrating these trees into statistical models or machine learning—are either unaware of these tools or find them difficult to use due to the steep learning curve of processing raw sequences. This is compounded by the fact that public datasets often lack phylogenetic trees, providing only abundance tables and taxonomic classifications. To address this, we present a comprehensive review of phylogenetic tree construction techniques for microbiome data (16S rRNA or whole-genome shotgun sequencing). We outline the strengths and limitations of current methods, offering expert insights and step-by-step guidance to make these tools more accessible and widely applicable in quantitative microbiome data analysis.

Dissecting AlphaFold2’s Capabilities with Limited Sequence Information

Abstract Protein structure prediction aims to infer a protein’s three dimensional structure from its amino acid sequence. Protein structure is pivotal for elucidating protein functions, interactions, and driving biotechnological innovation. The deep learning model AlphaFold2, has revolutionized this field by leveraging phylogenetic information from multiple sequence alignments to achieve remarkable accuracy in protein structure prediction. However, a key question remains: how well does AlphaFold2 understand protein structures? This study investigates AlphaFold2’s capabilities when relying primarily on high-quality template structures, without the additional information provided by multiple sequence alignments. By designing experiments that probe local and global structural understanding, we aimed to dissect its dependence on specific features and its ability to handle missing information. Our findings revealed AlphaFold2’s reliance on sterically valid Cβ for correctly interpreting structural templates. Additionally, we observed its remarkable ability to recover three dimensional structures from certain perturbations and the negligible impact of the previous structure in recycling. Collectively, these results support the hypothesis that AlphaFold2 has learned an accurate biophysical energy function. However, this function seems most effective for local interactions. Our work advances understanding of how deep learning models predict protein structures and provides guidance for researchers aiming to overcome limitations in these models.

A preliminary phylogeny of Malachiinae (Coleoptera, Malachiidae): From multiple genes to whole genomics

AbstractMalachiinae (Coleoptera: Malachiidae), commonly known as true soft‐winged flower beetles, are widely distributed across the globe except New Zealand. It is the largest subfamily of Cleroidea (over 3000 species) and is currently divided into seven tribes, among which the relationships are hard to assess due to their bizarre morphological modifications in males. Although a few molecular phylogenetics were attempted, they were all constructed on basis of four or five short nucleotide fragments, which may not contain enough phylogenetic information, thereby resulting in unclear relationships. To investigate the phylogenetic relationships among the tribes within Malachiinae, low‐coverage whole genomes of 26 species from the superfamily Cleroidea were sequenced and analysed, including 22 species of Malachiinae. The mitochondrial genomes (mitogenomes), nuclear genes (18S, 28S rRNA) and universal single‐copy orthologs (USCOs) were obtained as molecular markers from low‐coverage sequencing data. A total of 24 complete or nearly complete mitochondrial genomes of Malachiidae species were annotated and analysed. Further combined the newly sequenced data and those available in the public database, four different data matrices were analysed by different methods to reconstruct the phylogeny of Malachiinae. The results indicated that these phylogenetic analyses produced consistent topologies, and the phylogenetic relationships within Malachiinae were recovered as (Illopini + (Troglopini + (Colotini + Apalochrini))) + (Ebaeini + (Attalini + Malachiini)).

Phylogenetic Insights Into Canidae Trait Variation Across Continents

ABSTRACTAimUnderstanding the spatial structuring of ecological communities involves considering the interplay between evolutionary history and environmental factors. This study investigates how the phylogenetic structure of Canidae influences the geographical distribution and trait patterns of lineages globally, and how these patterns relate to Bergmann's and Rapoport's rules.LocationAmericas, Africa, Eurasia.Time Period12 million years ago—present.Major Taxa StudiedCanidae.MethodsUsing distribution data and phylogenetic information for 37 Canidae species, we analysed key ecological, functional and evolutionary variables. We applied phylogenetic fuzzy‐weighting via principal coordinates of phylogenetic structure (PCPS) and variance partitioning analysis (VPA) to assess the contributions of phylogenetic structure and environmental factors to trait variation among species.ResultsOur results revealed distinct global patterns in body size, body weight, range size, habitat use and evolutionary distinctiveness among lineages. We also identified the shared contributions of phylogenetic structure and temperature to trait variation using variance partitioning analysis. The PCPS axes highlighted the influence of phylogenetic relationships on Canidae assemblages, particularly in South America.Main ConclusionsImportantly, the study challenges the applicability of Bergmann's and Rapoport's rules across continents. The unique diversification history of Canidae in South America and Africa and their diverse environmental conditions likely contribute to the observed trait patterns that make both continents so distinguished when compared to North America and Eurasia. Our findings underscore the need to incorporate phylogenetic information in models assessing trait variation across geographic scales for unbiased estimates.

Ecological and taxonomic dissimilarity in species and higher taxa of reptiles in western Mexico.

Reptiles are one of the most diverse groups of vertebrates in the world that are distributed in almost all ecosystems. Many of these studies have focused on exploring their diversity patterns across different environments; and recent studies on reptile alpha and beta diversity have incorporated a multifaceted approach into their analysis to have more comprehensive evaluations. This study presents an assessment of the taxonomic diversity of reptile patterns using methods that incorporate the assessment of higher taxa. Likewise, the taxonomic dissimilarities between reptile communities in the physiographic regions of the state of Jalisco were analyzed. Evaluations for the groups of snakes and lizards independently are presented. We use the taxonomic distinctiveness index that assesses the complexity of the taxonomic structure of communities through hierarchical classification above the species level to measure the relationships between taxa. The dissimilarity of the taxonomic structure in each community was also analyzed. Beta diversity partitions were performed to identify the contribution of turnover and the differences in richness. We determined that alpha diversity of species and higher taxa maintain different patterns, indicating that Jalisco presents regions with overrepresentation of reptile families and genera, as well as regions with an opposite trend. The representation of higher taxa is higher in the lizard group, although in terms of species richness snakes are the most prominent group. The turnover is the most important component at species and higher taxa, with similar values for lizards and snakes. The findings presented show that incorporating phylogenetic information about species through taxonomic relationships provides complementary information that species diversity per se, especially at the level of alpha diversity.

Heritability in the Rhesus Macaque (Macaca mulatta) Vertebral Column.

The vertebral column plays a central role in primate locomotion and positional behavior. Understanding its evolution, therefore, has the potential to clarify evolutionary processes that have occurred in the primate lineage as well as the specific behaviors of extinct primates. However, to understand primate vertebral anatomy, it is important to determine how much of this anatomy is heritable and how much develops as a response to environmental factors during life. We estimated heritability for vertebral counts as well as typical cervical, thoracic, and lumbar elements from 210 individuals from the pedigreed Cayo Santiago Macaca mulatta skeletal collection. We found moderate heritability of vertebral counts (h2 = 0.216-0.326), but with strong heritability of the type of variation (e.g., a tendency to meristic or homeotic change) in the vertebral count (h2 = 0.599), suggesting a possible explanation for high variability in vertebral numbers among the hominoids in particular. The moderate heritability of vertebral count also suggests that vertebral count is an unsuitable metric for estimating the ancestral state for some taxa. We found strong heritability in the morphology of cervical and upper lumbar zygapophyseal facets (h2 = 0.548-0.550) and the thoracic spinous processes (h2 = 0.609-0.761), including high heritability of the spinous process angle in the upper thoracic and upper lumbar elements (h2 = 0.649-0.752). We suggest these are related to maintaining stability in the cervical and lumbar regions, and reducing motion in the thoracic region, respectively. We propose that spinous processes may contain greater phylogenetic information, whereas transverse processes may contain greater information of function 'in life'. We also found important size effects, suggesting that size is the most heritable component of overall form and largely responsible for intertrait differences. This suggests that it is inappropriate to indiscriminately remove size effects from morphological comparisons.

Hyperostosis in Fishes: An Update With New Species Records.

Literature reports of hyperostosis are often misleading and have been confused with osteomas, a pathological condition. Hyperostotic bones are known to occur only in bony fishes of the class Actinopterygii, within at least 16 orders, 35 families, 89 genera, and 153 species. They are present almost exclusively in marine fishes and exceptionally in a few extinct freshwater species known from hypersaline environments and one extant cichlid. Hyperostosis is best represented in the family Carangidae where it is known to occur in 53 of approximately 181 valid species. We also provide a synthetic report on what we know and what misconceptions exist regarding hyperostosis. Patterns of hyperostosis are often species-specific but provide no useful phylogenetic information. In species known to develop hyperostosis, it is usually not apparent (non-histologically) in juveniles and typically only becomes fully developed in the largest individuals. The timing of hyperostosis on-set in different bones is often sequential rather than simultaneous across different bones. Most marine Neoteleostei have acellular skeletons but histological observations have shown that in species exhibiting hyperostosis, areas of active remodeling are composed primarily of cellular bone characterized by a rich vascular network and bone-resorbing osteoclasts.

DeepPhylo: Phylogeny-Aware Microbial Embeddings Enhanced Predictive Accuracy in Human Microbiome Data Analysis.

Microbial data analysis poses significant challenges due to its high dimensionality, sparsity, and compositionality. Recent advances have shown that integrating abundance and phylogenetic information is an effective strategy for uncovering robust patterns and enhancing the predictive performance in microbiome studies. However, existing methods primarily focus on the hierarchical structure of phylogenetic trees, overlooking the evolutionary distances embedded within them. This study introduces DeepPhylo, a novel method that employs phylogeny-aware amplicon embeddings to effectively integrate abundance and phylogenetic information. DeepPhylo improves both the unsupervised discriminatory power and supervised predictive accuracy of microbiome data analysis. Compared to the existing methods, DeepPhylo demonstrates superiority in informing biologically relevant insights across five real-world microbiome use cases, including clustering of skin microbiomes, prediction of host chronological age and gender, diagnosis of inflammatory bowel disease (IBD) across 15 studies, and multilabel disease classification.

A New Dawn for Protist Biogeography

ABSTRACTAimBiogeographers have believed for a long time that the geographical distributions of protists are only determined by environmental conditions, because dispersal is not limited. During the past two decades, the field has come a long way to show that historical and spatial factors also significantly contribute to shaping protist distributions, calling for a reappraisal of our understanding of protist biogeography.MethodsWe review the current state‐of‐the‐art on the field of protist biogeography, highlighting several outstanding questions and opportunities. Our review brings together insights from different disciplines, ranging from morphology‐based research to environmental, population and speciation genomics.ResultsProtist communities harbour cosmopolitan and geographically restricted species and are shaped by both local environmental conditions and historical processes, yet the relative contributions of these patterns and processes likely differs depending on the geographic scale, protist lineage and the habitat that is being investigated. The field is ready to move beyond the decades‐long ubiquity versus (moderate) endemicity discourse and to instead ask why and where specific protist species and clades are more prone to widespread or restricted distributions. With the advent of next‐generation sequencing technologies, from whole‐genome sequencing to environmental and ancient DNA surveys, it is now possible to integrate insights from multiple lines of evidence and investigate protist communities, species and populations at an unprecedented scale and detail.OutlookTo further advance the field, the protist community needs to focus on understudied habitats and protist lineages, study the impact of protist traits on biogeographical patterns, perform targeted field and experimental work to disentangle the processes that underlie protist biogeographies and expand and develop databases with sequence, trait, distributional and phylogenetic information of protists. Given that a good understanding of species boundaries is central to unravelling protist biogeography, it remains crucial to invest in polyphasic taxonomic research.

Genotyping and phylogeographic dynamics of coxsackievirus A16

Coxsackievirus A16 (CV-A16) is one of the major pathogens of Hand, Foot and Mouth disease. Here, we analyzed 287 full-length genome sequences of CV-A16 found worldwide from 1994 to 2019 to see the genomic evolution characteristics. Full-length genome-based phylogenetic tree divided the viruses into five different genotypes, G-a to G-e. The CV-A16 strains circulating in China dominate G-a and G-c, but can also be found in other genotypes including G-b and G-e. Phylogeographic analysis showed a high diversity of CV-A16 distribution. In addition, recombination was shown to drive the genomic evolution of CV-A16 during past decades. However, the structural proteins still remain relative conserved while there is extensive genomic recombination. This study updates the phylogenetic and phylogeographic information of CV-A16 and provides insights into the genetic characteristics of CV-A16.

Microbacterium rhizophilus sp. nov., an indole acetic acid-producing actinobacterium isolated from rhizosphere soil.

A novel gram-stain-positive, short rod, aerobic, non-motile and non-spore-forming actinobacterial strain, designated GXG1230T was isolated from the rhizosphere soil of a coastal mangrove forest in Beihai city, Guangxi Zhuang Autonomous Region, PR China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain GXG1230T was affiliated with the genus Microbacterium. Additionally, it demonstrated a high degree of similarity to Microbacterium paludicola US15T (97.9%) and Microbacterium marinilacus YM11-607T (97.3%). Chemotaxonomic characteristics showed that the whole-cell sugars were glucose, xylose, rhamnose and galactose. Menaquinones MK-11 and MK-12 were detected as respiratory quinones. Lysine was found in the peptidoglycan hydrolysate and the polar lipids were diphosphatidylglycerol, one phospholipid and two unidentified glycolipid. The major fatty acids were anteiso-C15:0, iso-C16:0 and anteiso-C17:0. The strain GXG1230T exhibited a genomic DNA G + C content of 71.7%. Furthermore, the average nucleotide identity values of GXG1230T with the reference strains were 75.4% and 81.9%, respectively, while the digital DNA-DNA hybridization values were 20.1% and 25.0%. Based on physiological, chemotaxonomic and phylogenetic information, strain GXG1230T is considered to represent a novel species of the genus Microbacterium, for which the name Microbacterium rhizophilus sp.nov is proposed, with GXG1230T (= MCCC 1K09302T = KCTC 59252T) as the type strain.

Rational selection of TbpB variants elucidates a bivalent vaccine formulation with broad spectrum coverage against Neisseria gonorrhoeae.

Neisseria gonorrhoeae is the causative agent of gonorrhea, an on-going public health problem due in part to the lack of success with efforts to develop an efficacious vaccine to prevent this sexually transmitted infection. An attractive candidate vaccine antigen because of its essential function and surface exposure, the gonococcal transferrin binding protein B (TbpB) exhibits high levels of antigenic variability which poses a significant obstacle in evoking a broadly protective vaccine composition. Here, we utilize phylogenetic information to rationally select TbpB variants for inclusion into a potential gonococcal vaccine and identify two TbpB variants that when formulated together elicit a highly cross-reactive antibody response in both rabbits and mice against a diverse panel of TbpB variants and clinically relevant gonococcal strains. Further, this formulation performed well in experimental proxies of real-world usage, including eliciting bactericidal activity against 8 diverse gonococcal strains and decreasing the median duration of colonization after vaginal infection in female mice by two heterologous strains of N. gonorrhoeae . Together, these data support the use of a combination of TbpB variants for a broadly protective gonococcal vaccine.

Developing Novel Microsatellite Markers for Kaempferia parviflora by Microsatellite Capture Sequencing (MiCAPs)

Kaempferia parviflora, a medicinal plant widely used in Southeast Asia, has been validated clinically for its diverse pharmaceutical applications. Despite extensive research in pharmacology, there is a notable lack of cytogenetic and genomic research, primarily due to limited genetic information. Simple Sequence Repeat (SSR) is considered a robust class of molecular markers frequently used in biodiversity studies. In this study, we adopted Microsatellite Capture Sequencing (MiCAPs) to obtain SSR sequences for marker development. We identified 13,644 SSRs and developed and validated ten sets of SSR markers through capillary electrophoresis. The ten primer sets generated 27 alleles, with an average Polymorphism Information Content (PIC) of 0.36. Principle Coordinate Analysis (PCoA) distinguished two types of K. parviflora, consistent with classification by leaf margin color (red and green). A neighbor-joining dendrogram of seven Zingiberaceae species was constructed with the SSR-containing sequences. The 2-c value of K. parviflora is first reported here as 3.16 ± 0.03; the genome size is estimated at 3090.48 Mbp. The newly developed molecular markers are crucial for variety identification and the conservation of wild resources. Additionally, the cytogenetic and phylogenetic information provides valuable insights into the genetic diversity and evolutionary relationships.

StackbarExtended: a user-friendly stacked bar-plot representation incorporating phylogenetic information and microbiota differential abundance analysis

Background Microbial communities are mainly composed of bacteria, archaea, viruses and fungi, and are present in the gut, mouth, nose, skin, lungs, vagina, and bladder, among other places. In recent years, research has highlighted the critical role that these highly complex communities play in health and disease. Advances in sequencing technology have resulted in the development of high-dimensional data, which are challenging to effectively analyze and visualize. In this context, traditional stacked bar-plot visualizations, while widely used, fall short of conveying the fundamental phylogenic relationships between community members and are thus difficult to interpret. Methods StackbarExtended is implemented in native R, required version (≥ 4.0), and is platform independent, with its source code available on GitHub and archived on Zenodo. Results StackbarExtended allows for the plotting of relative abundance at user-defined taxonomic levels while displaying phylogenetic information using color gradients. Additionally, StackbarExtended integrates differential abundance statistics directly into the visualization process and performs clustering of low-abundance taxa. Conclusions StackbarExtended offers researchers a user-friendly tool for rapid visualization, presentation, and analysis of the microbiota composition.

Phylogeny structures species' interactions in experimental ecological communities.

Species' traits and interactions are products of evolutionary history. Despite the long-standing hypothesis that closely related species possess similar traits, and thus experience stronger competition, measuring the effect of evolutionary history on the ecology of natural communities remains challenging. We propose a novel framework to test whether phylogeny influences patterns of coexistence and abundance of species assemblages. In our approach, phylogenetic trees are used to parameterize species' interactions, which in turn determine the abundance of species in a given assemblage. We use likelihoods to score models parameterized with a given phylogeny, and contrast them with models built using random trees, allowing us to test whether phylogenetic information helps to predict species' abundances. Our statistical framework reveals that interactions are indeed structured by phylogeny in a large set of experimental plant communities. Our results confirm that evolutionary history can help predict, and potentially manage or conserve, the structure and function of complex ecological communities.

A permutable MLP-like architecture for disease prediction from gut metagenomic data

Metagenomic data plays a crucial role in analyzing the relationship between microbes and diseases. However, the limited number of samples, high dimensionality, and sparsity of metagenomic data pose significant challenges for the application of deep learning in data classification and prediction. Previous studies have shown that utilizing the phylogenetic tree structure to transform metagenomic abundance data into a 2D matrix input for convolutional neural networks (CNNs) improves classification performance. Inspired by the success of a Permutable MLP-like architecture in visual recognition, we propose Metagenomic Permutator (MetaP), which applied the Permutable MLP-like network structure to capture the phylogenetic information of microbes within the 2D matrix formed by phylogenetic tree. Our experiments demonstrate that our model achieved competitive performance compared to other deep neural networks and traditional machine learning, and has good prospects for multi-classification and large sample sizes. Furthermore, we utilize the SHAP (SHapley Additive exPlanations) method to interpret our model predictions, identifying the microbial features that are associated with diseases.

Comparative osteology of the skull of cowbirds (Icteridae: Molothrus).

Detailed osteological descriptions of the craniomandibular complex of passerine birds are lacking for most species, limiting our understanding of their diversity and evolution. Cowbirds (genus Molothrus) are a small but widespread group of New World nine-primaried songbirds, well-known for their unique brooding parasitic behavior. However, detailed osteological data for cowbirds and other Icteridae are currently scarce and several features of their skulls remain undescribed or poorly known. To address this issue, a detailed comparative osteology of cowbird skulls is presented here for the first time based on data from x-ray microcomputed tomography, dry skeletal data, and multivariate analyses of linear morphometric data. Cowbird skulls offer some functional insights, with many finch-like features probably related to a seed-rich diet that distinguishes them from most other icterids. In addition, features previously overlooked in earlier studies might provide valuable phylogenetic information at different levels of passerine phylogeny (Passerida, Emberizoidea, Icteridae, and Agelaiinae), including some of the otic region and nasal septum. Comparisons among cowbirds show that there is substantial cranial variation within the genus, with M. oryzivorus being the most divergent cowbird species. Within the genus, distantly related species share similar overall skull morphology and proportions, but detailed osteological data allow species identification even in cases of strong convergence. Further efforts are warranted to furnish baseline data for future studies of this iconic group of Neotropical birds and to fully integrate it into phylogenetic comparative frameworks.

The future of plant diversity within a Mediterranean endemism centre: Modelling the synergistic effects of climate and land-use change in Peloponnese, Greece

Climate- and land-use change stand as primary threats to terrestrial biodiversity. Yet, their synergistic impacts on species distributions remain poorly understood. To address this knowledge gap, we conducted the first-ever comprehensive species distribution analysis on an entire regional endemism centre within an eastern Mediterranean country, incorporating dynamic land-use/land-cover change data together with climate change scenarios. Specifically, we apply species distribution modelling and spatial data analysis techniques to compare the individual and synergistic effects of these environmental drivers on the endemic vascular flora of Peloponnese, focusing on potential range contractions, altitudinal shifts, and habitat fragmentation levels. Moreover, we identify fine-scale present and potential future endemism hotspots within our study area, incorporating taxonomic and phylogenetic information. Overall, we aim to enhance our current understanding of endemism patterns and contribute to the development of future-proof conservation strategies for safeguarding Greece's endangered endemic flora. The integration of land-use change projections with climate change yielded less severe impacts compared to the effects anticipated when considering climatic variables alone. Most taxa are expected to undergo significant range declines and nearly half might experience increased habitat fragmentation, due to the synergistic effects of climate- and land-use change. We identified endemism hotspots, which are concentrated in or along the main Peloponnesian mountain massifs. However, our predictions indicate that areas presently recognized as endemism hotspots will undergo a concerning area decline, across all future scenarios considered in this study. Our findings highlight the importance of including dynamic land-use variables alongside climatic predictors when projecting species distributions under global change. Moreover, we showed that endemism hotspots are not static and considering their potential geographic shifts is paramount to delineate effective forward-looking conservation strategies.

Molecular based identification and phylogenetic relationship of the leech Hirudinaria manillensis from India by using mitochondrial cytochrome c oxidase subunit I gene.

A molecular approach for the identification of unknown species by the using mitochondrial cox1 gene is an effective and reliable as compared with morphological-based identification. Hirudinaria manillensis referred to as Asian Buffalo Leech, is found in South Asia and traditionally used as medicine owing to its medicinal properties. The study aimed to isolate and identify the leech species using cox1 gene sequencing and their phylogenetic relationships. The nucleotide sequences of cytochrome c oxidase subunit I (cox1) mitochondrial genes were analyzed for species identification and the phylogenetic relationship of crucial therapeutic leech Hirudinaria manillensis. The isolated DNA from the leech sample was amplified with cox1 gene-specific primers. BLAST results with the H. manillensis sequence showed 89.24% homology with H. manillensis and phylogenetic tree analysis revealed the genetic relationship with other GenBank submitted sequences. The present study concluded that the cox1 gene could be an effective way to identify the leech H. manillensis and provided sufficient phylogenetic information to distinguish H. manillensis indicating a significant mtDNA-based approach to species identification.