Comparative Phylogenetic Analysis Research Articles

The reduced ARF regulatory system in Giardia intestinalis pre-dates the transition to parasitism in the lineage Fornicata

Giardia intestinalis is an enteric pathogen with an extremely modified membrane trafficking system, lacking canonical compartments such as the Golgi, endosomes, and intermediate vesicle carriers. By comparison the fornicate relatives of Giardia possess greater endomembrane system complexity. In eukaryotes, the ADP ribosylation factor (ARF) GTPase regulatory system proteins, which consist of the small GTPase ARF1, and its guanine exchange nucleotide factors (GEFs) and GTPase activating proteins (GAPs), coordinate temporal and directional trafficking of cargo vesicles by recognizing and interacting with heterotetrameric coat complexes at pre-Golgi and post-Golgi interfaces. To understand the evolution of this regulatory system across the fornicate lineage, we have performed comparative genomic and phylogenetic analyses of the ARF GTPases, and their regulatory GAPs and GEFs in fornicate genomes and transcriptomes. Prior to our analysis of the fornicates, we first establish that the ARF GAP sub-family ArfGAP with dual PH domains (ADAP) is sparsely distributed but present in at least four eukaryotic supergroups and thus was likely present in the Last Eukaryotic Common Ancestor (LECA). Next, our collective comparative genomic and phylogenetic investigations into the ARF regulatory proteins in fornicates identify a duplication of ARF1 GTPase yielding two paralogues of ARF1F proteins, ancestral to all fornicates and present in all examined isolates of Giardia. However, the ARF GEF and ARF GAP complement is reduced compared with the LECA. This investigation shows that the system was significantly streamlined prior to the fornicate ancestor but was not further reduced concurrent with a transition into a parasitic lifestyle.

Ecological Correlates of 20-Year Population Trends of Wintering Waterbirds in Deep Bay, South China

Along the East Asian-Australasian flyway (EAAF), waterbirds are threatened by a wide range of human activities. Studies have shown that wintering populations of many species have declined in Australia and Japan; however, long term data along China’s coast are limited. In this study, we analyzed data collected from monthly bird surveys to quantify population trends of wintering waterbirds from 1998 to 2017 in the Deep Bay area, South China. Of the 42 species studied, 12 declined, while nine increased significantly. Phylogenetic comparative analysis revealed that population trends were negatively correlated to reliance on the Yellow Sea and body size. Further, waterbird species breeding in Southern Siberia declined more than those breeding in East Asia. These findings, coupled with a relatively high number of increasing species, support the continual preservation of wetlands in the Deep Bay area. This study provides another case study showing that data collected from wintering sites provide insights on the threats along migratory pathway and inform conservation actions. As such, we encourage population surveys in the EAAF to continue, particularly along the coast of China.

Fossil pollen from early Palaeogene sediments in western India provides phylogenetic insights into divergence history and pollen character evolution in the pantropical family Ebenaceae

AbstractMany plant families lack substantive fossil records, limiting our understanding of their origin and evolution. The abundance and preservation potential of pollen through geological time have helped to overcome such limitations and have provided reliable fossils for reconstructing biogeographical history and character evolution in many angiosperm families. Here, using scanning electron microscopy, we identified six Ebenaceae-type fossil pollen grains from early Palaeogene sediments of western India. Phenetic and phylogenetic analyses using pollen characters of fossil and extant taxa reavealed affinities of these fossils to three genera of Ebenaceae (Euclea, Royena and Diospyros). Furthermore, our divergence dating analysis using these fossils as priors suggested a Gondwanan origin for the family during the mid-Cretaceous [c. 107 Mya, 95% highest posterior density (HPD): 100–112 Mya] and supports the boreotropical and ‘out of India’ dispersal hypotheses as the most probable explanations for the present global distribution of the family. The study also supports the dispersal of the family into India, from Africa, through the Kohistan–Ladakh Arc during the Palaeocene. Finally, comparative phylogenetic analyses suggest significant synapomorphic and phylogenetic signals for a few selected pollen characters in Ebenaceae. Our findings have important implications for understanding the biogeography and evolution of the highly diverse and ecologically and economically important family Ebenaceae.

Sequence and Structural Characterization of Toll-Like Receptor 6 from Human and Related Species.

Toll-like receptors (TLRs) play an important role in the innate immune response against various pathogens. They serve as expected targets of natural selection in those species which are adapted to habitats with contrasting pathogen burdens. Till date, sufficient literature about TLRs especially TLR6 is not available. The current study was therefore planned to show evolutionary patterns of human TLRs generally and TLR6 specifically along with their conservation and diversity. The study also deals with characteristic polymorphic patterns of TLR6 in humans which are involved in serious clinical consequences. The sequence analysis of TLR6 from different mammals revealed conserved regions in the protein sequence. With respect to TLR6 evolution, human showed a close evolutionary relationship with chimpanzee and orangutans, while monkeys were appeared in a separate clade showing a distant evolutionary relationship. Old World monkeys and New World monkeys made their separate clades but both have evolved from a common ancestor. The C-terminal of human TLRs (TLR1 to TLR10) exhibited more conservation as compared to other regions. The phylogram of human TLRs showed that TLR6 is closely related to TLR1 and both TLRs shared a common ancestor with TLR10. The domain analysis has revealed that TLR1 and TLR10 have least (i.e., 4) number of leucine-rich repeat (LRR) while TLR6 contains five LRRs. Three single nucleotide polymorphisms were found in TLR6 which were found to be associated with benign. Conclusively, the current comparative sequence analyses and phylogenetic analyses provided informative insights into the process of TLR evolution in mammals. Furthermore, the polymorphism analysis would serve as a useful marker in the early detection of susceptibility and resistance against cancers and other diseases in humans.

Trade‐offs in expressed major histocompatibility complex diversity seen on a macroevolutionary scale among songbirds

To survive organisms must defend themselves against pathogens. Classical Major Histocompatibility Complex (MHC) genes play a key role in pathogen defense by encoding molecules involved in pathogen recognition. MHC diversity influences the variety of pathogens individuals can recognize and respond to and has consequently been a popular genetic marker for disease resistance in ecology and evolution. However, MHC diversity is predominantly estimated using genomic DNA (gDNA) with little knowledge of expressed diversity. This limits our ability to interpret the adaptive significance of variation in MHC diversity, especially in species with very many MHC genes, such as songbirds. Here we address this issue using phylogenetic comparative analyses of the number of MHC class I alleles (MHC-I diversity) in gDNA and complementary DNA (cDNA), i.e. expressed alleles, across thirteen songbird species. We propose three theoretical relationships that could be expected between genomic and expressed MHC-I diversity on a macro-evolutionary scale and test which of these are best supported. In doing so, we show that significantly fewer MHC-I alleles than the number available are expressed, suggesting that optimal MHC-I diversity could be achieved by modulating gene expression. Understanding the relationship between genomic and expressed MHC-I diversity is essential for interpreting variation in MHC diversity in an evolutionary context. This article is protected by copyright. All rights reserved.

Evolutionary implications of dental anomalies in bats.

The gain or loss of anatomical features is an important mechanism of morphological evolution and ecological adaptation. Dental anomalies-the loss or gain of teeth-are widespread and a potential source of craniodental specialization among mammals, yet their macroevolutionary patterns have been rarely explored. We present the first phylogenetic comparative study of dental anomalies across the second largest mammal Order, Chiroptera (bats). We conducted an extensive literature review and surveyed a large sample of museum specimens to analyze the types and prevalence of dental anomalies across bats, and performed phylogenetic comparative analyses to investigate the role of phylogenetic history and dietary specialization on incidence of dental anomalies. We found dental anomalies have a significant phylogenetic signal, suggesting they are not simply the result of idiosyncratic mutations or random developmental disorders, but may have ancestral genetic origins or result from shared developmental pathways among closely related species. The incidence of dental anomalies was not associated with diet categories, suggesting no effect of craniodental specialization on dental anomalies across bats. Our results give insight into the macroevolutionary patterns of dental anomalies in bats, and provide a foundation for investigating new hypotheses underlying the evolution of dental variation and diversity in mammals.

Personality of hosts and their brood parasites.

Brood parasites such as the common cuckoo Cuculus canorus exploit the parental abilities of their hosts, hosts avoid brood parasitism and predation by showing specific behavior such as loss of feathers, emission of fear screams and contact calls, displaying wriggle behavior to avoid hosts or potential prey, pecking at hosts and prey, and expressing tonic immobility (showing behavior like feigning death or rapid escape from predators and brood parasites). These aspects of escape behavior are consistent for individuals but also among sites, seasons, and years. Escape behavior expressed in response to a broad range of cuckoo hosts and prey are consistently used against capture by humans, but also hosts and brood parasites and predators and their prey. An interspecific comparative phylogenetic analysis of escape behavior by hosts and their brood parasites and prey and their predators revealed evidence of consistent behavior when encountering potential parasites or predators. We hypothesize that personality axes such as those ranging from fearfulness to being bold, and from neophobic to curiosity response in brood parasites constitute important components of defense against brood parasitism that reduces the overall risk of parasitism.

The non-gradual nature of adaptive radiation

Adaptive radiation is a major source of biodiversity. Still, many aspects of this evolutionary process remain poorly understood. Our recent integrative examination of the cichlid adaptive radiation in African Lake Tanganyika provides new insights into the process of explosive diversification. The in-depth phylogenetic comparative analysis of nearly all species occurring in that lake permitted us to trace patterns of eco-morphological evolution throughout the phylogenetic history of the radiation and revealed that it occurred in a non-gradual manner, in the form of time-shifted bursts of accelerated evolution. The temporal pattern of divergence along different axes of morphological evolution provides empirical support for a scenario that potentially unifies two popular models of adaptive radiation, the “early burst model” and the “stages model”.

Characterization and Full Genome Sequence of Novel KPP-5 Lytic Phage against Klebsiella pneumoniae Responsible for Recalcitrant Infection.

Klebsiella pneumoniae is a hazardous opportunistic pathogen that is involved in many serious human diseases and is considered to be an important foodborne pathogen found in many food types. Multidrug resistance (MDR) K. pneumoniae strains have recently spread and increased, making bacteriophage therapy an effective alternative to multiple drug-resistant pathogens. As a consequence, this research was conducted to describe the genome and basic biological characteristics of a novel phage capable of lysing MDR K. pneumoniae isolated from food samples in Egypt. The host range revealed that KPP-5 phage had potent lytic activity and was able to infect all selected MDR K. pneumoniae strains from different sources. Electron microscopy images showed that KPP-5 lytic phage was a podovirus morphology. The one-step growth curve exhibited that KPP-5 phage had a relatively short latent period of 25 min, and the burst size was about 236 PFU/infected cells. In addition, KPP-5 phage showed high stability at different temperatures and pH levels. KPP-5 phage has a linear dsDNA genome with a length of 38,245 bp with a GC content of 50.8% and 40 predicted open reading frames (ORFs). Comparative genomics and phylogenetic analyses showed that KPP-5 is most closely associated with the Teetrevirus genus in the Autographviridae family. No tRNA genes have been identified in the KPP-5 phage genome. In addition, phage-borne virulence genes or drug resistance genes were not present, suggesting that KPP-5 could be used safely as a phage biocontrol agent.

Chloroplast genomes of two Mediterranean Bupleurum species and the phylogenetic relationship inferred from combined analysis with East Asian species.

The chloroplast genomes of Mediterranean Bupleurum species are reported for the first time. Phylogenetic analysis supports the species as a basal clade of Bupleurum with divergence time at 35.40Ma. Bupleurum is one of the most species-rich genus with high medicinal value in Apiaceae. Although infrageneric classifications of Bupleurum have been the subject of numerous studies, it still remains controversial. Chloroplast genome information will prove essential in advancing our understanding on phylogenetic study. Here we report cp genomes of two woody Bupleurum species (Bupleurum gibraltaricum and B. fruticosum) endemic to Mediterranean. The complete cp genomes of the two species were 157,303 and 157,391bp in size, respectively. They encoded 114 unique genes including 30 tRNA genes, 4 rRNA genes and 80 protein coding genes. Genome structure, distributions of SDRs and SSRs, gene content exhibited similarities among Bupleurum species. High variable hotspots were detected in eight intergenic spacers and four genes. Most of genes were under purifying selection with two exceptions: atpF and clpP. The phylogenetic analysis based on 80 coding genes revealed that the genus was divided into 2 distinct clades corresponding to the 2 subgenera (subg. Penninervia, subg. Bupleurum) with divergence time at the end of collision of India with Eurasia. Most species diversified mainly during the later period of uplift of Qinghai-Tibetan Plateau. The cp genomes of the two Bupleurum species can be significant complementary to insights into the cp genome characteristics of this genus. The comparative chloroplast genomes and phylogenetic analysis advances our understanding of the evolution of cp genomes and phylogeny in Bupleurum.

Genomic Characterization and Distribution Pattern of a Novel Marine OM43 Phage.

Bacteriophages have a significant impact on the structure and function of marine microbial communities. Phages of some major bacterial lineages have recently been shown to dominate the marine viral communities. However, phages that infect many important bacterial clades still remained unexplored. Members of the marine OM43 clade are methylotrophs that play important roles in C1 metabolism. OM43 phages (phages that infect the OM43 bacteria) represent an understudied viral group with only one known isolate. In this study, we describe the genomic characterization and biogeography of an OM43 phage that infects the strain HTCC2181, designated MEP301. MEP301 has a genome size of 34,774 bp. We found that MEP301 is genetically distinct from other known phage isolates and only displays significant sequence similarity with some metagenomic viral genomes (MVGs). A total of 12 MEP301-type MVGs were identified from metagenomic datasets. Comparative genomic and phylogenetic analyses revealed that MEP301-type phages can be separated into two subgroups (subgroup I and subgroup II). We also performed a metagenomic recruitment analysis to determine the relative abundance of reads mapped to these MEP301-type phages, which suggested that subgroup I MEP301-type phages are present predominantly in the cold upper waters with lower salinity. Notably, subgroup II phages have an inverse different distribution pattern, implying that they may infect hosts from a distinct OM43 subcluster. Our study has expanded the knowledge about the genomic diversity of marine OM43 phages and identified a new phage group that is widespread in the ocean.

What drives species’ distributions along elevational gradients? Macroecological and ‐evolutionary insights from Brassicaceae of the central Alps

AbstractAimGeographic distribution limits of organisms are often affected by climate, but little is known of how the impacts of climate evolve within sets of related taxa. Here we identified the climate variables most closely associated with low‐elevation limits, optimal elevations, and high‐elevation limits of plant species’ distributions and compared evolutionary lability of niche values predicting the three aspects of distribution best.LocationCentral Alps.Time periodCurrent.Major taxa studiedThe plant family Brassicaceae.MethodsWe modelled the occurrence of 110 brassicaceous species in the central European Alps and used response curves of predicted occurrence on climatic variables to reveal those variables most strongly associated with elevational distribution. We produced a phylogeny of the species, applied phylogenetic comparative analysis and tested whether niche values predicting the low and high limits and the optimum of elevational distribution were similar among related taxa.ResultsUpper limits were closely associated with the length of the vegetation season for the majority of species, while summer or spring temperatures were strongly allied with both the occurrence optimum and the lower limit. Furthermore, niche values predicting the upper limit and the optimum of elevational distribution were less conserved in contrast to niche values predicting the lower limit of distribution.Main conclusionsThese results highlight constraints on adaptation at the warm end of the climate niche and may explain observed range retractions at warm range edges due to ongoing climate change.

Evolutionary Sample Size and Consilience in Phylogenetic Comparative Analysis.

Phylogenetic comparative methods (PCMs) are commonly used to study evolution and adaptation. However, frequently used PCMs for discrete traits mishandle single evolutionary transitions. They erroneously detect correlated evolution in these situations. For example, hair and mammary glands cannot be said to have evolved in a correlated fashion because each evolved only once in mammals, but a commonly used model (Pagel's Discrete) statistically supports correlated (dependent) evolution. Using simulations, we find that rate parameter estimation, which is central for model selection, is poor in these scenarios due to small effective (evolutionary) sample sizes of independent character state change. Pagel's Discrete model also tends to favor dependent evolution in these scenarios, in part, because it forces evolution through state combinations unobserved in the tip data. This model prohibits simultaneous dual transitions along branches. Models with underlying continuous data distributions (e.g., Threshold and GLMM) are less prone to favor correlated evolution but are still susceptible when evolutionary sample sizes are small. We provide three general recommendations for researchers who encounter these common situations: i) create study designs that evaluate a priori hypotheses and maximize evolutionary sample sizes; ii) assess the suitability of evolutionary models-for discrete traits, we introduce the phylogenetic imbalance ratio; and iii) evaluate evolutionary hypotheses with a consilience of evidence from disparate fields, like biogeography and developmental biology. Consilience plays a central role in hypothesis testing within the historical sciences where experiments are difficult or impossible to conduct, such as many hypotheses about correlated evolution. These recommendations are useful for investigations that employ any type of PCM. [Class imbalance; consilience; correlated evolution; evolutionary sample size; phylogenetic comparative methods.].

Host specialization and molecular evidence support a distinct species of smut fungus, Anthracoidea hallerianae (Anthracoideaceae), on Carex halleriana (Cyperaceae)

The species of Anthracoidea on Carex are host-specific smut fungi restricted to sedges belonging to the same or closely related sections. They are characterized by sori that form black, hard bodies around aborted nuts of their hosts. In Carex sect. Hallerianae, only one species, C. halleriana, is known as a host of Anthracoidea. The taxonomic status of this smut fungus was problematic due to a lack of molecular data. It has been reported under different names, mainly as “A. caricis” or “A. irregularis”. A comparative morphological study and molecular phylogenetic analysis, using LSU (large subunit) nuclear rDNA sequences, supported the recognition of a distinct species, A. hallerianae. The new species is described and illustrated based on material from Central Europe, the Iberian Peninsula, the Balkan Peninsula, the Mediterranean area and Transcaucasia.Citation: Denchev T. T., Denchev C. M., Koopman J., Begerow D. & Kemler M. 2021: Host specialization and molecular evidence support a distinct species of smut fungus, Anthracoidea hallerianae (Anthracoideaceae), on Carex halleriana (Cyperaceae). – Willdenowia 51: 57–67.Version of record first published online on 9 March 2021 ahead of inclusion in April 2021 issue.

Is degree of sociality associated with reproductive senescence? A comparative analysis across birds and mammals.

Our understanding on how widespread reproductive senescence is in the wild and how the onset and rate of reproductive senescence vary among species in relation to life histories and lifestyles is currently limited. More specifically, whether the species-specific degree of sociality is linked to the occurrence, onset and rate of reproductive senescence remains unknown. Here, we investigate these questions using phylogenetic comparative analyses across 36 bird and 101 mammal species encompassing a wide array of life histories, lifestyles and social traits. We found that female reproductive senescence: (i) is widespread and occurs with similar frequency (about two-thirds) in birds and mammals; (ii) occurs later in life and is slower in birds than in similar-sized mammals; (iii) occurs later in life and is slower with an increasingly slower pace of life in both vertebrate classes; and (iv) is only weakly associated, if any, with the degree of sociality in both classes after accounting for the effect of body size and pace of life. However, when removing the effect of species differences in pace of life, a higher degree of sociality was associated with later and weaker reproductive senescence in females, which suggests that the degree of sociality is either indirectly related to reproductive senescence via the pace of life or simply a direct outcome of the pace of life. This article is part of the theme issue 'Ageing and sociality: why, when and how does sociality change ageing patterns?'

Selection for functional performance in the evolution of cuticle hardening mechanisms in insects.

Calcified tissues have repeatedly evolved in many animal lineages and show a tremendous diversity of forms and functions. The cuticle of many insects is enriched with elements other than Calcium, a strategy of hardening that is taxonomically widespread but apparently poorly variable among clades. Here, we investigate the evolutionary potential of the enrichment with metals in insect cuticle at different biological levels. We combined experimental evidence of Zinc content variation in the mandibles of a target species (Chorthippus cazurroi [Bolívar]) with phylogenetic comparative analyses among grasshopper species. We found that mandibular Zinc content was repeatable among related individuals and was associated with an indicator of fitness, so there was potential for adaptive variation. Among species, Zinc enrichment evolved as a consequence of environmental and dietary influences on the physical function of the jaw (cutting and chewing), suggesting a role of natural selection in environmental fit. However, there were also important within and transgenerational environmental sources of similarity among individuals. These environmental influences, along with the tight relationship with biomechanics, may limit the potential for diversification of this hardening mechanism. This work provides novel insights into the diversification of biological structures and the link between evolutionary capacity and intra- and interspecific variation.

Comparative bioacoustics: a roadmap for quantifying and comparing animal sounds across diverse taxa.

Animals produce a wide array of sounds with highly variable acoustic structures. It is possible to understand the causes and consequences of this variation across taxa with phylogenetic comparative analyses. Acoustic and evolutionary analyses are rapidly increasing in sophistication such that choosing appropriate acoustic and evolutionary approaches is increasingly difficult. However, the correct choice of analysis can have profound effects on output and evolutionary inferences. Here, we identify and address some of the challenges for this growing field by providing a roadmap for quantifying and comparing sound in a phylogenetic context for researchers with a broad range of scientific backgrounds. Sound, as a continuous, multidimensional trait can be particularly challenging to measure because it can be hard to identify variables that can be compared across taxa and it is also no small feat to process and analyse the resulting high-dimensional acoustic data using approaches that are appropriate for subsequent evolutionary analysis. Additionally, terminological inconsistencies and the role of learning in the development of acoustic traits need to be considered. Phylogenetic comparative analyses also have their own sets of caveats to consider. We provide a set of recommendations for delimiting acoustic signals into discrete, comparable acoustic units. We also present a three-stage workflow for extracting relevant acoustic data, including options for multivariate analyses and dimensionality reduction that is compatible with phylogenetic comparative analysis. We then summarize available phylogenetic comparative approaches and how they have been used in comparative bioacoustics, and address the limitations of comparative analyses with behavioural data. Lastly, we recommend how to apply these methods to acoustic data across a range of study systems. In this way, we provide an integrated framework to aid in quantitative analysis of cross-taxa variation in animal sounds for comparative phylogenetic analysis. In addition, we advocate the standardization of acoustic terminology across disciplines and taxa, adoption of automated methods for acoustic feature extraction, and establishment of strong data archival practices for acoustic recordings and data analyses. Combining such practices with our proposed workflow will greatly advance the reproducibility, biological interpretation, and longevity of comparative bioacoustic studies.

Genome Sequence and Characterization of Acinetobacter Phage DMU1.

Background: There has been a recent resurgence of research on the characterization of Acinetobacter phage for therapeutic use due to the morbidity and mortality associated with treatment failures in cases of multidrug-resistant Acinetobacter baumannii infections. Materials and Methods: A bacteriophage isolated from activated sludge that targets A. baumannii ATCC19606 was characterized by electron microscopy, genome sequencing, comparative genomics, and a host range analysis. Results: The morphology of Acinetobacter phage DMU1 resembles phages in Siphoviridae. Comparative genomic and phylogenetic analyses reveal that DMU1 is a siphophage and is most closely related to Acinetobacter phage SH-Ab 15497. Out of the strains tested, DMU1 was found to only infect A. baumannii strains ATCC19606 and ATCC17978. Conclusion: Acinetobacter phage DMU1 belongs to the Siphoviridae family and is most closely related to Acinetobacter phage SH-Ab 15497. Small-scale host-range analysis of DMU1 indicates a host range that is likely limited to specific A. baumannii strains.

A novel vieuvirus from multidrug-resistant Acinetobacter baumannii.

Bacteriophages are considered the most abundant biological entities on earth, and they are able to modulate the populations of their bacterial hosts. Although the potential of bacteriophages has been accepted as an alternative strategy to combat multidrug-resistant pathogenic bacteria, there still exists a considerable knowledge gap regarding their genetic diversity, which hinders their use as antimicrobial agents. In this study, we undertook a genomic and phylogenetic characterization of the phage Ab11510-phi, which was isolated from a multidrug-resistant Acinetobacter baumannii strain (Ab11510). We found that Ab11510-phi has a narrow host range and belongs to a small group of transposable phages of the genus Vieuvirus that have only been reported to infect Acinetobacter bacteria. Finally, we showed that Ab11510-phi (as well as other vieuvirus phages) has a high level of mosaicism. On a broader level, we demonstrate that comparative genomics and phylogenetic analysis are necessary tools for the proper characterization of phage diversity.

Biodiversity of New Lytic Bacteriophages Infecting Shigella spp. in Freshwater Environment.

Bacteriophages, viruses that infect and replicate within prokaryotic cells are the most abundant life forms in the environment, yet the vast majority of them have not been properly reported or even discovered. Almost all reported bacteriophages infecting the Enterobacteriaceae family, with Escherichia coli being the major subject of studies, have been isolated from wastewater, sewage, and effluent resources. In the present study, we focused on the distribution and biodiversity of Shigella phages in an aquatic ecosystem. While no Shigella bacteria was recovered from the Yangtze River, three lytic phages were isolated from this ecosystem and were subjected to biological, morphological, and genomic characteristics. Comparative genomics and phylogenetic analyses demonstrated that vB _SflM_004 isolate belongs to Myoviridae family, Felixounavirus genus of Ounavirinae subfamily, vB_SdyM_006 was classified under the same family, however, it is suggested to be in a new genus under Tevenvirinae subfamily with some other related bacteriophages. vB_SsoS_008 phage belongs to the Siphoviridae family, Tunavirus genus, Tunavirinae subfamily. The phages did not harbor any genes involved in the lysogenic cycles and showed a high temperature and pH stability. The biodiversity of the isolated phages highly suggests that continued isolation on non-model members of Enterobacteriaceae family is necessary to fully understand bacteriophage diversity in aquatic environments.