Phylogeny of elasmobranchs based on LSU and SSU ribosomal RNA genes

The dominant view of the phylogeny of living elasmobranchs, based on morphological characters, is that batoids (skates and rays) are derived sharks, joined with saw sharks, and angel sharks in the clade Hypnosqualea [S. Shirai, Squalean Phylogeny: A New Framework of 'Squaloid' Sharks and Related Taxa, Hokkaido University Press, Sapporo, 1992]. By contrast, a recent molecular-phylogenetic study based on mitochondrial genes for 12S and 16S rRNA and tRNA valine [C.J. Douady et al., Mol. Phylogenet. Evol., 26 (2003) 215-221] supported the older view that batoids and sharks are separate lineages. Here, we tested these two different views using combined, nuclear large-subunit and small-subunit rRNA gene sequences ( approximately 5.3kb) from 22 elasmobranchs, two chimeras, and two bony fishes. We used maximum likelihood, maximum parsimony, minimum evolution, and Bayesian inference for tree reconstruction, and found the large-subunit rRNA gene to contain far more signal than the small-subunit gene for resolving this mostly Mesozoic radiation. Our findings matched those of in separating batoids from sharks and in statistically rejecting Hypnosqualea. The angel shark (Squatina) was the sister group to squaliforms (dogfish sharks), and our findings are consistent with the idea that "orbitostylic" sharks form a monophyletic group (squaliforms+the hexanchiform Chlamydoselachus+Squatina+Pristiophorus). In the galeomorph sharks, however, lamniforms grouped with orectolobiforms, opposing the widely accepted 'lamniform+carcharhiniform' grouping. A tree based on the mitochondrial gene for cytochrome b also supported a separation of sharks and batoids, in contrast to Hypnosqualea. Among elasmobranchs, variation in the evolutionary rates of the nuclear rRNA genes was higher than that of cytochrome b genes, mainly due to the relatively rapid evolution of rRNA in some carcharhiniforms. In conclusion, several different molecular studies now refute the Hypnosqualea hypothesis of elasmobranch interrelationships.     

Higher elasmobranch phylogeny and biostratigraphy

Living sharks, skates and rays share several derived skeletal characters that are absent in most extinct elasmobranchs, suggesting a monophyletic group of 'higher' elasmobranchs. Within this group opinions vary as to phylogenetic relationships, although three broad groups are generally recognized. Arguments for and against monophyly of these group (batoids; squalomorphs; galeomorphs) are examined. Many of their contained taxa are also of questionable validity. Cladistic analysis of living galeomorphs reveals a sequence of characters supporting monophyly of the group as whole, but not of its more generalized contained taxa. The temporal distribution of fossil galeomorphs corroborates the hypothesis of relationship suggested by neontological data; i.e. there is considerable stratigraphic harmony with Recent phylogenetic data.     

Molecular phylogenetic convergence within Elasmobranchii revealed by cytochrome oxidase subunits

The phylogenetic relationships within the subclass of Elasmobranchii are under question within the academic community and their systematic classification based on morphological, or physiological characteristics has not yet been fully justified. Modern cladistic studies suggested that batoids are derived sharks, a taxonomic status known as the Hypnosqualean hypothesis. The main purpose of this study was to address this issue using a data set of aligned, directly sequenced, mitochondrial cytochrome oxidase subunits I and II. Maximum Likelihood, Maximum Parsimony, Minimum Evolution and Bayesian inference were implemented for tree reconstructions. The results provided evidence that supported the rejection of the above hypothesis, in accordance with other recent molecular phylogenetic studies. More specifically Rajiformes species were presented as separate lineages from sharks. Prionace species on the other hand was grouped within Carcharhinoformes, which was clustered as sister group to Lamniformes. COI and COII regions supported, monophylies of Squaliformes and paraphylies of Carchariniformes.     

Shark tales: a molecular species-level phylogeny of sharks (Selachimorpha, Chondrichthyes)

Sharks are a diverse and ecologically important group, including some of the ocean's largest predatory animals. Sharks are also commercially important, with many species suffering overexploitation and facing extinction. However, despite a long evolutionary history, commercial, and conservation importance, phylogenetic relationships within the sharks are poorly understood. To date, most studies have either focused on smaller clades within sharks, or sampled taxa sparsely across the group. A more detailed species-level phylogeny will offer further insights into shark taxonomy, provide a tool for comparative analyses, as well as facilitating phylogenetic estimates of conservation priorities. We used four mitochondrial and one nuclear gene to investigate the phylogenetic relationships of 229 species (all eight Orders and 31 families) of sharks, more than quadrupling the number of taxon sampled in any prior study. The resulting Bayesian phylogenetic hypothesis agrees with prior studies on the major relationships of the sharks phylogeny; however, on those relationships that have proven more controversial, it differs in several aspects from the most recent molecular studies. The phylogeny supports the division of sharks into two major groups, the Galeomorphii and Squalimorphii, rejecting the hypnosqualean hypothesis that places batoids within sharks. Within the squalimorphs the orders Hexanchiformes, Squatiniformes, Squaliformes, and Pristiophoriformes are broadly monophyletic, with minor exceptions apparently due to missing data. Similarly, within Galeomorphs, the orders Heterodontiformes, Lamniformes, Carcharhiniformes, and Orectolobiformes are broadly monophyletic, with a couple of species 'misplaced'. In contrast, many of the currently recognized shark families are not monophyletic according to our results. Our phylogeny offers some of the first clarification of the relationships among families of the order Squaliformes, a group that has thus far received relatively little phylogenetic attention. Our results suggest that the genus Echinorhinus is not a squaliform, but rather related to the saw sharks, a hypothesis that might be supported by both groups sharing 'spiny' snouts. In sum, our results offer the most detailed species-level phylogeny of sharks to date and a tool for comparative analyses.     

Morphology and phylogenetic significance of the pectoral articular region in elasmobranchs (Chondrichthyes)

The morphology of the articular region of the scapulocoracoid and the basal cartilages of the pectoral fin endoskeleton of elasmobranchs is reviewed in detail. Examination of this specific morphology in more than 140 species of elasmobranchs (of which 40 are reported here) revealed characters that may have a bearing on the higher‐level phylogeny of the group. Ten distinct characters of the scapular articular region of elasmobranchs are described, varying in terms of the number of distinct articular sites as well as their specific morphology (e.g. whether the articular surface is composed of condyles and/or facets). Previous interpretations of the articular region in morphological phylogenies are also reviewed, revealing much more morphological variation than formerly reported. These prior characters played an important role in supporting the Hypnosqualea, and may still be derived for this clade. The variation and distribution of the new characters discussed provide new insights for the evolution of the pectoral endoskeleton in chondrichthyans. They also highlight the continued importance of morphological characters for phylogenetic studies, and reinforce the necessity of in‐depth anatomical reviews of certain characters employed in previous higher‐level phylogenetic studies of elasmobranchs. © 2015 The Linnean Society of London     

Molecular phylogeny of elasmobranchs inferred from mitochondrial and nuclear markers

The elasmobranchs (sharks, rays and skates) being the extant survivors of one of the earliest offshoots of the vertebrate evolutionary tree are good model organisms to study the primitive vertebrate conditions. They play a significant role in maintaining the ecological balance and have high economic value. Due to over-exploitation and illegal fishing worldwide, the elasmobranch stocks are being decimated at an alarming rate. Appropriate management measures are necessary for restoring depleted elasmobranch stocks. One approach for restoring stocks is implementation of conservation measures and these measures can be formulated effectively by knowing the evolutionary relationship among the elasmobranchs. In this study, a total of 30 species were chosen for molecular phylogeny studies using mitochondrial cytochrome c oxidase subunit I, 12S ribosomal RNA gene and nuclear Internal Transcribed Spacer 2. Among different genes, the combined dataset of COI and 12S rRNA resulted in a well resolved tree topology with significant bootstrap/posterior probabilities values. The results supported the reciprocal monophyly of sharks and batoids. Within Galeomorphii, Heterodontiformes (bullhead sharks) formed as a sister group to Lamniformes (mackerel sharks): Orectolobiformes (carpet sharks) and to Carcharhiniformes (ground sharks). Within batoids, the Myliobatiformes formed a monophyly group while Pristiformes (sawfishes) and Rhinobatiformes (guitar fishes) formed a sister group to all other batoids.     

Factors affecting species richness of marine elasmobranchs

Many studies on elasmobranchs, sharks and batoids (rays, skates and guitarfishes), have focused on the factors responsible for biomass decline, but little attention has been paid to the factors that affect species richness. We used the software package ModestR to determine the geographical distribution of all valid marine elasmobranch species (512 species of sharks and 619 species of batoids), thereby making it possible to determine the species composition of the elasmobranch community in any area worldwide. The primary aim of this study was to identify the factors associated with the species richness of elasmobranchs. The data were analyzed using multiple regressions and Support Vector Machine (SVM) in cells of 1º× 1º with the analyzed abiotic variables being bathymetry, chlorophyll a, sea surface temperature, photosynthetically available radiation, pH, cloud cover, the concentrations of calcite, silicate, phosphate and nitrate, salinity, particulate organic carbon, diffuse attenuation and dissolved oxygen. The mean area of occupancy of the species was used as an indicator of niche occupancy. The model performed with SVM explained 97 and 99 % of the variance observed in the species richness of batoids and sharks, respectively. Mean area of occupancy, temperature and bathymetry were the variables with a higher contribution to the variance observed in both sharks and batoids. The negative residuals of the model performed with SVM indicated areas with lower than predicted species richness. These may be potential areas with undiscovered and/or unregistered species, or areas with decreased species richness due to the negative effect of anthropogenic factors, i.e. overfishing     

Review of Elasmobranch Behavioral Studies Using Ultrasonic Telemetry with Special Reference to the Lemon Shark, Negaprion Brevirostris, Around Bimini Islands, Bahamas

A review of past behavioral ultrasonic telemetry studies of sharks and rays is presented together with previously unpublished material on the behavior of the lemon shark, Negaprion brevirostris, around the Bimini Islands, Bahamas. The review, focusing on movement behaviors of 20 shark and three ray species, reveals that elasmobranchs exhibit a variety of temporal and spatial patterns in terms of rates-of-movement and vertical as well as horizontal migrations. The lack of an apparent pattern in a few species is probably attributable to the scarcity of tracking data. Movements are probably governed by several factors, some still not studied, but data show that food, water temperature, bottom type, and magnetic gradient play major roles in a shark's decision of where and when to swim. A few species exhibit differences in behavior between groups of sharks within the same geographical area. This interesting finding warrants further research to evaluate the causes of these apparent differences and whether these groups constitute different subpopulations of the same species. The lack of telemetry data on batoids and some orders of sharks must be addressed before we can gain a more comprehensive understanding of the behavior of elasmobranch fishes. Previously unpublished data from 47 smaller and 38 larger juvenile lemon sharks, collected over the decade 1988–1998, provide new results on movement patterns, habitat selection, activity rhythms, swimming speed, rate-of-movement, and homing behavior. From these results we conclude that the lemon shark is an active predator with a strong, apparently innate homing mechanism. This species shows ontogenetic differences in habitat selection and behavior, as well as differences in movements between groups of individuals within the same area. We suggest three hypotheses for future research on related topics that will help to understand the enigmatic behavior of sharks.     

The elasmobranchs of Malpelo Flora and Fauna Sanctuary,Colombia

Management and conservation actions in marine-protected areas require baselines for monitoring threatened marine fauna such as elasmobranchs. This article provides evidence of the occurrence of 34 species of elasmobranchs (21 sharks and 13 batoids) in the Malpelo Flora and Fauna Sanctuary, Colombia, including five new records of sharks and three of rays. From 1987 to 2021, new records were obtained by underwater visual census using SCUBA, manned submersibles and deep-ocean cameras to depths of up to 2211 m. Of the recorded species, 21 are considered as threatened taxa (64%) by the IUCN, making the Malpelo Flora and Fauna Sanctuary an essential conservation area for this highly threatened group of species.     

Determinants of tapeworm species richness in elasmobranch fishes: untangling environmental and phylogenetic influences

Parasite species richness is a fundamental characteristic of host species and varies substantially among host communities. Hypotheses aiming to explain observed patterns of richness are numerous, and none is universal. In this study, we use tapeworm parasites of elasmobranch fishes to examine the phylogenetic and environmental influences on the variation in species richness for this specific system. Tapeworms are the most diverse group of helminths to infect elasmobranchs. Elasmobranchs are cosmopolitan in distribution and their tapeworm parasites are remarkably host specific; therefore, making this an ideal system in which to examine global patterns in species diversity. Here, we 1) quantify the tapeworm richness in elasmobranch fishes, 2) identify the host features correlated with tapeworm richness, and 3) determine whether tapeworm richness follows a latitudinal gradient. The individual and combined effects of host size, factors associated with water temperatures (influenced by latitude and depth), host habitat, and type of elasmobranch (shark or batoid) on measures of species diversity were assessed using general linear models. These analyses included tapeworm host records for 317 different elasmobranch species (124 species were included in our analyses) and were conducted with and without taking into account phylogenetic relationships between host species. Since sharks and batoids differ substantially in body form, analyses were repeated for each host subset. On average, batoids harboured significantly more tapeworm species than shark hosts. Tapeworm richness in sharks was influenced by median depth, whereas no predictor variable included in our models could adequately account for interspecific variation in tapeworm richness in batoid hosts. The taxonomic diversity of tapeworm assemblages of sharks and batoids was influenced by median depth and median latitude, respectively. When the influence of host phylogeny is accounted for, larger hosts harbour a greater tapeworm richness, whereas hosts exploiting wider latitudinal ranges harbour more taxonomically distinct tapeworm assemblages. Species richness and taxonomic diversity of tapeworm assemblages in elasmobranch fishes are influenced by different evolutionary pressures, including host phylogenetic relationships, space constraints and geographical area. Our results suggest that ca 3600 tapeworm species have yet to be described from elasmobranch fishes.     

Pectoral fin morphology of batoid fishes (Chondrichthyes: Batoidea): Explaining phylogenetic variation with geometric morphometrics

The diverse cartilaginous fish lineage, Batoidea (rays, skates, and allies), sister taxon to sharks, comprises a huge range of morphological diversity which to date remains unquantified and unexplained in terms of evolution or locomotor style. A recent molecular phylogeny has enabled us to confidently assess broadscale aspects of morphology across Batoidea. Geometric morphometrics quantifies the major aspects of shape variation, focusing on the enlarged pectoral fins which characterize batoids, to explore relationships between ancestry, locomotion and habitat. A database of 253 specimens, encompassing 60 of the 72 batoid genera, reveals that the majority of morphological variation across Batoidea is attributable to fin aspect‐ratio and the chordwise location of fin apexes. Both aspect‐ratio and apex location exhibit significant phylogenetic signal. Standardized independent linear contrast analysis reveals that fin aspect‐ratio can predict locomotor style. This study provides the first evidence that low aspect‐ratio fins are correlated with undulatory‐style locomotion in batoids, whereas high aspect‐ratio fins are correlated with oscillatory locomotion. We also show that it is phylogeny that determines locomotor style. In addition, body‐ and caudal fin‐locomotors are shown to exhibit low aspect‐ratio fins, whereas a pelagic lifestyle correlates with high aspect‐ratio fins. These results emphasize the importance of phylogeny in determining batoid pectoral fin shape, however, interactions with other constraints, most notably locomotor style, are also highlighted as significant. J. Morphol. 275:1173–1186, 2014. © 2014 Wiley Periodicals, Inc.     

Monophyly, phylogeny and systematic position of the †Synechodontiformes (Chondrichthyes, Neoselachii)

Klug, S. (2009). Monophyly, phylogeny and systematic position of the †Synechodontiformes (Chondrichthyes, Neoselachii). — Zoologica Scripta, 39 , 37–49.
Identifying the monophyly and systematic position of extinct sharks is one of the major challenges in reconstructing the phylogeny and evolutionary history of sharks in general. Although great progress has been accomplished in the last few decades with regard to resolving the interrelationships of living sharks, a comprehensive phylogeny identifying the systematic position of problematic or exclusively fossil taxa is still lacking. Fossil taxa traditionally assigned to synechodontiform sharks are very diverse with a fossil record extending back into the Palaeozoic but with uncertain inter- and intrarelationships. Here, phylogenetic analyses using robust cladistic principles are presented for the first time to evaluate the monophyly of this group, their intrarelationships and their systematic position within Neoselachii. According to the results of this study, taxa assigned to this group form a monophyletic clade, the †Synechodontiformes. This group is sister to all living sharks and displays a suite of neoselachian characters. Consequently, the concept of neoselachian systematics needs to be enlarged to include this completely extinct group, which is considered to represent stem-group neoselachians. The origin of modern sharks can be traced back into the Late Permian (250 Mya) based on the fossil record of †Synechodontiformes. The systematic position of batoids remains contradictory, which relates to the use of different data (molecular vs. morphological) in phylogentic analyses.     

An investigation of the co-evolutionary relationships between onchobothriid tapeworms and their elasmobranch hosts

There is general consensus that the living elasmobranchs comprise a monophyletic taxon. There is evidence that, among tetraphyllidean tapeworms, the approximately 201 hooked species (Onchobothriidae) may also comprise a monophyletic group. Determinations of host specificity are contingent upon correct specific identifications. Since 1960, over 200 new elasmobranch species and over 100 new onchobothriid species have been described. Some confidence can be placed in host and parasite identifications of recent studies, but specific identifications provided in older literature in many cases are suspect. There is some consensus among published works on the phylogenetic relationships among elasmobranchs. Phylogenetic relationships among onchobothriids remain largely unresolved. Elasmobranchs have been poorly sampled for onchobothriids; records exist for approximately 20% of the 911 species and approximately 44% of the 170 elasmobranch genera. Onchobothriids are remarkably host specific, exhibiting essentially oioxenous specificity for their definitive hosts. Multiple onchobothriid species commonly parasitise the same host species; in some cases these are congeners, in other cases these are members of two different onchobothriid genera. There is substantial incongruence between available host and parasite phylogenies. For example, Acanthobothrium is by far the most ubiquitous onchobothriid genus, parasitising almost all orders of elasmobranchs known to host onchobothriids, yet, there is no evidence of major clades of Acanthobothrium corresponding to postulated major subgroupings of elasmobranchs (e.g. Galea and Squalea or sharks and rays). Potamotrygonocestus appears to be among the most basal onchobothriid groups, yet it parasitises one of the most derived elasmobranch groups (the freshwater stingray genus Potamotrygon). It appears that congeners parasitising the same host species are not necessarily each other's closest relatives. At this point the preliminary and limited available data suggest that, at least in this system, strict host specificity is not necessarily indicative of strict co-evolution. This study was extremely limited by the lack of available robust phylogenies for onchobothriids and elasmobranchs.     

The neural control of feeding in elasmobranchs: A review and working model

A working model of the neural control of feeding in elasmobranchs is presented and summarized in graphic form. The model is based on a review of studies in sharks and batoids augmented by suggestions and comparisons from research in mammals and teleosts. The focal point of the model is a proposed Hypothalamic Feeding Area (HFA) that encompasses the medial periventricular zone in the inferior lobe and a small area immediately dorsal to it. Electrical stimulation in the HFA has evoked feeding in nurse sharks and neuropeptides and neurotransmitters known to influence feeding in mammals and teleosts have been localized immunocytochemically in the region in several elasmobranchs. The HFA of elasmobranchs appears to be analogous to and possibly homologous with ??hypothalamic feeding centers?? in bony fishes and tetrapods. Such ??centers?? are thought to integrate external and internal stimuli and control feeding in relation to available energy stores. The HFA??s strong olfactory connections in elasmobranchs are consistent with smell-induced feeding activities. In elasmobranchs, the HFA has reciprocal connections with the central pallium of the telencephalon, a region that processes visual, acoustic, mechanoreceptive and electroreceptive lateral line and possibly somatosensory information. These pathways may provide multisensory control in feeding. HFA connections with the cerebellum, brainstem and spinal cord most likely mediate hypothalamic co-ordination of the sensorimotor components of elasmobranch feeding. The review and model help to identify areas for suggested research.     

How can the feeding habits of the sand tiger shark influence the success of conservation programs?

The feeding habits of the sand tiger shark Carcharias taurus , one of the most threatened sharks of the world, are poorly known. Sand tiger sharks are critically endangered in the South-west Atlantic. Since 2007, the law requires that all individuals caught in recreational fisheries off Argentina must be released. Using data from a north Patagonian recreational fishery ( n =164 stomachs with contents), we analyzed the diet of sand tiger sharks in relation with size, sex, maturity stage and season; assessed prey consumption patterns and hooking location; and estimated diet overlap with fishery landings. Sand tiger sharks consumed mainly teleosts (55.4% of the total prey number, N ) and elasmobranchs (41.84% N ), and ate more benthic elasmobranchs (batoids and angel sharks) as they become larger. Sharks swallowed prey mostly in one piece (93.7%) and were hooked mainly in internal organs (87.4%, n =175), causing occlusion and perforation of the esophagus and stomach, and lacerations to the pericardium, heart and liver. Sand tiger sharks fed on the most heavily landed species, overlapping almost completely (>90%) with fishery landings. Conservation plans should take into account that releasing hooked sharks could be insufficient to minimize fishing mortality and that competition for food with fisheries is likely to occur.     

Phylogenetic relationships among microgastrine braconid wasp genera based on data from the 16S, COI and 28S genes and morphology

Abstract Phylogenetic relationships among the genera of the large braconid wasp subfamily Microgastrinae were explored using DNA sequence data from the mitochondrial large ribosomal subunit (16S), nuclear large ribosomal subunit (28S) and mitochondrial cytochrome oxidase (COI) genes, along with morphological characters, both new and from previous studies. The taxonomic history of this group of wasps is reviewed, along with a critique of previous phylogenetic studies on the group. Molecular data were sampled from forty-six species representing twenty-six genera of microgastrines, plus three species representing the close outgroup taxa Cardiochilinae and Miracinae. Some 2300 base pairs of aligned sequence were obtained per taxon from the three genes. In addition, fifty-three morphological characters were coded for all known genera, including two undescribed genera, except Semionis Nixon (known from only a single male type specimen). Relationships among several groups of genera are clarified and challenge some major assumptions made in earlier classifications. In particular, it is clear that dependence on one or a few major morphological character systems oversimplifies relationships, and can lead to misleading results. Despite the large amount of data analysed, basal divergences within the subfamily remain poorly resolved and essentially unsupported in any rigorous statistical sense.     

Molecular tests of phylogenetic taxonomies: a general procedure and example using four subfamilies of the lizard family Iguanidae

A general procedure is described for examining when results of molecular phylogenetic analyses warrant formal revision of taxonomies constructed using morphological characters. We illustrate this procedure with tests of monophyly for four subfamilies in the lizard family Iguanidae using 1561 aligned base positions (838 phylogenetically informative) of mitochondrial DNA sequences, representing coding regions for eight tRNAs, ND2, and portions of ND1 and COI. Ten new sequences ranging in length from 1732 to 1751 bases are compared with 12 previously reported sequences and 67 morphological characters (54 phylogenetically informative) from the literature. New morphological character states are provided for Sator. Phylogenies derived from the molecular and combined data are in agreement but both conflict with phylogenetic inferences from the morphological data alone. Strong support is found for the monophyly of the subfamilies Crotaphytinae and Phrynosomatinae. Monophyly of the Iguaninae is weakly supported in each analysis. All analyses suggest that the Tropidurinae is not monophyletic but the hypothesis of monophyly cannot be rejected. A phylogenetic taxonomy is proposed in which the Tropidurinae* is maintained as a metataxon (denoted with an asterisk), for which monophyly has not been demonstrated. Within the Phrynosomatinae, the close relationship of Sator and Sceloporus is questioned and an alternative hypothesis in which Sator is the sister taxon to a clade comprising Petrosaurus, Sceloporus, and Urosaurus is presented. Statistical tests of monophyly provide a powerful way to evaluate support for taxonomic groupings. Use of the metataxon prevents premature taxonomic rearrangements where support is lacking.     

Proopiomelanocortin (POMC) and testing the phylogenetic position of turtles (Testudines)

Morphological and molecular studies have inferred multiple hypotheses for the phylogenetic relationships of Testudines. The hypothesis that Testudines are the only extant anapsid amniotes and the sister taxon of diapsid amniotes is corroborated by morphological studies, while the hypothesis that Testudines are diapsid amniotes is corroborated by more recent molecular and morphological studies. In this study, the placement of Testudines is tested using the full length cDNA sequence of the polypeptide hormone precursor proopiomelanocortin (POMC). Because only extant taxa have been used, the hypotheses being tested are limited to the following (1) Testudines as the sister taxon of Archosauria, (2) Testudines included in Archosauria and the sister taxon of Crocodilia, (3) Testudines as the sister taxon of Lepidosauria, (4) Testudines as the sister taxon of Sauria, and (5) Testudines as the sister taxon of a monophyletic Mammalia–Sauria clade. Neither Maximum likelihood, Bayesian, or maximum parsimony analyses are able to falsify the hypothesis of (Archosauria (Lepidosauria, Testudines)) and as such is the preferred inference from the POMC data.