Consistent seasonal flexibility of the gut and its regions across wild populations of a winter-quiescent fish.

Seasonality in north-temperate environments imposes drastic temperature and resource variations that shape the seasonal ecophysiology of resident organisms. A better understanding of an organism's capacity to flexibly respond to this drastic seasonal variation may reveal important mechanisms for tolerating or responding to environmental variation introduced by global change. In fishes, the digestive system is both the interface between resource and energy acquisition and one of the most expensive organ systems to maintain. However, little evidence describing the capacity for seasonal flexibility in the digestive tract of wild northern fishes exists. Here, we investigated phenotypic flexibility in the size of the gastrointestinal (GI) tract across three northern populations of a winter-dormant warm-water fish, pumpkinseed sunfish (Lepomis gibbosus). In all populations, pumpkinseed exhibited pronounced structural flexibility in the GI tract, aligned with winter and the timing of reproduction. The dry mass of the GI increased by 1.3- to nearly 2.5-fold in the early spring. The pyloric caeca demonstrated the greatest capacity for flexibility, increasing by up to 3.7-fold prior to reproduction. In all populations, minimum dry GI mass was consistently achieved during winter and mid-summer. This capacity for gut flexibility may represent a novel mechanism for facilitating rapid adaptive responses (e.g. metabolic plasticity) to future environmental change.

Mitochondrial Enzyme Activities and Body Condition of Naturally Infected Sunfish (Lepomis gibbosus).

AbstractParasites can affect host behavior, cognition, locomotion, body condition, and many other physiological traits. Changes to host aerobic metabolism may be responsible for these parasite-induced performance alterations. Whole-organism metabolic rate is underpinned by cellular energy metabolism driven most prominently by mitochondria. However, few studies have explored how mitochondrial enzymatic activity relates to body condition and parasite infection, despite it being a putative site for metabolic disruptions related to health status. We studied correlations among natural parasite infection, host body condition, and activity of key mitochondrial enzymes in target organs from wild-caught pumpkinseed sunfish (Lepomis gibbosus) to better understand the cellular responses of fish hosts to endoparasite infection. Enzymatic activities in the gills, spleen, and brain of infected fish were not significantly related to parasite infection or host body condition. However, the activity of cytochrome c oxidase, an enzyme involved in oxidative phosphorylation, in fish hearts was higher in individuals with a lower body condition. Activities of citrate synthase, electron transport system (complexes I and III), and carnitine palmitoyltransferase were also significantly different among organ types. These results provide preliminary information regarding the likely mitochondrial pathways affecting host body condition, the maintenance energetic requirements of different organs, and the organs' specific dependency on particular mitochondrial pathways. These results help pave the way for future studies on the effects of parasite infection on mitochondrial metabolism.

The cichlid pharyngeal jaw novelty enhances evolutionary integration in the feeding apparatus.

The modified pharyngeal jaw system of cichlid fishes is widely viewed as a key innovation that substantially facilitated the evolutionary exuberance of this iconic evolutionary radiation. We conduct comparative phylogenetic analyses of integration, disparity, and rate of evolution among feeding-related, skeletal structures in Neotropical cichlids and North American centrarchids, which lack the specialized pharyngeal jaw. Contrasting evolutionary patterns in these two continental radiations, we test a classic decoupling hypothesis. Specifically, we ask whether the modified pharyngeal jaw in cichlids resulted in enhanced evolutionary independence of the oral and pharyngeal jaws, leading to increased diversity of trophic structures. Contrary to this prediction, we find significantly stronger evolutionary integration between the oral and pharyngeal jaws in cichlids compared to centrarchids, although the two groups do not differ in patterns of integration within each jaw system. Further, though we find no significant differences in disparity, centrarchids show faster rates of morphological evolution. Our results suggest that the modified pharyngeal jaw resulted in less evolutionary independence and slower rates of evolution within the feeding system. Thus, we raise the possibility that the cichlid novelty enhances feeding performance, but does not prompt increased morphological diversification within the feeding apparatus, as has long been thought.

A reference genome for Bluegill (Centrarchidae: Lepomis macrochirus).

North American sunfishes (Family Centrarchidae) are among the most popular sportfish throughout the United States and Canada. Despite the popularity of sunfishes, their ecological importance, and their extensive stocking and aquacultural history, few molecular studies have examined the evolutionary relationships and species boundaries among members of this group, many of which are known to hybridize. Here, we describe a chromosome-scale genome assembly representing Bluegill (Lepomis macrochirus), one of the most widespread centrarchid species. By combining long-read, Oxford Nanopore sequencing data with short-insert, whole-genome and HiC sequence reads, we produced an assembly (Lm_LA_1.1) having a total length of 889 Mb including 1,841 scaffolds and having a scaffold N50 of 36 Mb, L50 of 12, N90 of 29 Mb, and L90 of 22. We detected 99% (eukaryota_odb10) and 98% (actinopterygii_odb10) universal single-copy orthologs (BUSCOs), and ab initio gene prediction performed using this new assembly identified a set of 17,233 genes that were supported by external (OrthoDB v10) data. This new assembly provides an important addition to the growing set of assemblies already available for spiny-rayed fishes (Acanthomorpha), and it will serve as a resource for future studies that focus on the complex evolutionary history of centrarchids.

New records of Spotted Bass, Micropterus punctulatus, within the Mississippi River Basin, Illinois.

Spotted Bass Micropterus punctulatus, like many sport fishes, have experienced range expansion through intentional introductions (i.e., legal stocking and illegal transfers) and migration across the United States. In Illinois, USA, native populations of Spotted Bass occur along the eastern and southern border of the state. We report new records of Spotted Bass in their non-native range of the Illinois Waterway and the Illinois portion of the Upper Mississippi River in addition to collections in their native range in the Illinois sections of the Ohio and Wabash rivers to better understand their current distribution. Continuous, collaborative efforts to track the distribution and expansion of non-native fishes are important for maintaining and establishing native and non-native fisheries management objectives and education, as non-native fishes can influence native species population distribution and dynamics.

Population genetic structure, parasite infection and somatic condition of pumpkinseed Lepomis gibbosus (Actinopterygii: Centrarchidae) in the Oder river basin.

In Poland, distribution of non-native pumpkinseed Lepomis gibbosus (Centrarchidae) is strictly limited to the Oder river basin, where it was introduced in the early 20th century. Recently, several populations have been found in waterbodies adjacent to the Oder, particularly in its lower reaches. In this study, we compare the genetic relatedness of populations in the Oder basin with other European populations using nuclear (microsatellite) and mitochondrial (partial cytochrome c oxidase subunit I; cox1) markers. Microsatellite analysis indicated that four populations in the lower Oder form a separate cluster, while one in the middle Oder clustered with Danubian populations, from where probably having been introduced. Microsatellite data suggested that the lower Oder populations differ from other non-native European populations, making it impossible to estimate the source of introduction. Nevertheless, analysis of cox1 indicated that Oder pumpkinseeds belong to the same haplotype as the vast majority of European populations. Parasitological examination confirmed the presence of two North American species, the monogenean Onchocleidus dispar and trematode Posthodiplostomum centrarchi, in the lower Oder, both previously unknown in the region. Fifteen other parasite species were acquired, including glochidia of invasive Sinanodonta woodiana. In the middle Oder, parasite infection was more limited. Fish from the Gryfino Canal, considered one of the most invasive populations in Europe, showed the highest parasite abundance and diversity, and the highest somatic condition and growth rate due to warm water released from the Dolna Odra power plant. Our results highlight significant differences in somatic condition and parasite infection in long-established non-native pumpkinseed populations in the same river system, reflecting mainly environmental conditions.

Two new species of Myxobolus (Myxozoa: Myxobolidae) infecting the gill and scales of the smallmouth bass, Micropterus dolomieu (Centrarchiformes: Centrarchidae) in the French Broad River Basin, North Carolina.

Two new species of Myxobolus Bütschli, 1882 (Bivalvulida: Myxobolidae) are described from the gill and scales of smallmouth bass (Micropterus dolomieu Lacepède, 1802 [Centrarchiformes: Centrarchidae]) from the Watauga River, French Broad River Basin, North Carolina, United States. Myxobolus intralamina n. sp. infects the lumen of the lamellar arterioles and Myxobolus infrabractea n. sp. infects the inner surface of the scale. They differ from all congeners by a combination of myxospore dimensions, polar tubule coil count, and the presence or absence of an iodinophilic vacuole in the sporoplasm and an intercapsular process. A phylogenetic analysis of the small subunit ribosomal DNA (SSU rDNA) recovered M. intralamina n. sp. sister to Myxobolus lepomis and Myxobolus branchiarum and M. infrabractea n. sp. sister to Myxobolus micropterii in a clade composed of five Myxobolus spp. infecting centrarchids and Henneguya spp. (Myxobolidae) infecting percids. Histological sections of infected gill revealed intra-lamellar plasmodia of M. intralamina n. sp. within the lumen of the lamellar arterioles and plasmodia of M. infrabractea n. sp. developing beneath the scales. These new species comprise the first species of Myxobolus reported from a black bass (Micropterus Lacepède, 1802) in the Southeast United States.

A generalized approach for sperm cryopreservation in the genus Pomoxis: Sperm cryopreservation and fertilization efficiency of black-stripe black crappie, Pomoxis nigromaculatus.

Approaches for white crappie, Pomoxis annularis sperm cryopreservation have led to interest in applying similar methods to black-stripe black crappie, Pomoxis nigromaculatus. Their rarity in wild populations makes them a preferred phenotype for hatchery use. Sperm cryopreservation procedures were compared between black-stripe black crappie and white crappie for sperm motility and egg fertilization rate. There was no difference in black-stripe black crappie sperm motility after thawing between 5% dimethyl sulfoxide (DMSO, 45% motility) and 10% methanol (50% motility). However, fertilization rates were higher (p < .001) for sperm cryoprotected with 5% DMSO (38 ± 8%) than 10% methanol (22 ± 7%). Hatchery use requires sperm-to-egg ratios and fertilizing potential of single doses (i.e., 0.5 ml straw). Using black-stripe black crappie sperm (2.5 × 108 sperm/ml; 5% DMSO), the highest fertilization (27%) was found using single straws with 785 eggs (0.25 ml); total sperm:egg ratio: 159,000:1; motile sperm:egg ratio: 71,700:1. Therefore, sperm of two Pomoxis species could be cryopreserved using 350 mOsmol/kg Hanks' balanced salt solution as an extender, 5% DMSO as a cryoprotectant, cooling at 40°C/min, and thawing for 8 s at 40°C to maintain sperm motility and fertility. Basic protocols can be generalized within a genus if variables such as sperm concentration, process timing, and sample volumes are controlled.

Morphology, phylogenetics and pathology of "red sore disease" (coinfection by Epistylis cf. wuhanensis and Aeromonas hydrophila) on sportfishes from reservoirs in the South-Eastern United States.

The aetiological agents of red sore disease (RSD) reportedly comprise a taxonomically ambiguous stalked ciliate (a species of Epistylis) and Aeromonas hydrophila. The taxonomic identity of each pathogen remains provisional: using supra-specific morphological features for the ciliate and culture-based methods that cannot delineate bacterial strain. On 7 and 9 November 2017 and 28 May 2020, biologists and anglers reported a local epizootic (Hiwassee and Chattahoochee river basins; Georgia) wherein some moribund fish presented RSD-like lesions. The ciliates were assigned to Epistylis by morphology. The ciliate is regarded as Epistylis cf wuhanensis, as nucleotide sequences from its small subunit ribosomal DNA were identical to those of Epistylis wuhanensis. The bacterium was identified as Aeromonas hydrophila by phenotypic markers and nucleotide sequences from the DNA gyrase subunit B; our sequences comprised 3 strains and phylogenetically were recovered sister to strains of Eurasian origin. Histological sections of lesions revealed effacement or partial deterioration of the epithelium covering scales, scale loss, haemorrhaging, necrosis, oedema, and extensive inflammatory infiltrate in the dermis. This is the first nucleotide sequence information for the symbionts implicated in RSD.

The Proteocephalus Species-Aggregate in Freshwater Centrarchid and Percid Fishes of the Nearctic Region (North America).

In the present paper, species of the Proteocephalus-aggregate de Chambrier, Zehnder, Vaucher, and Mariaux, 2004 (Cestoda: Proteocephalidae) reported from centrarchid and percid fishes in North America are reviewed, and their taxonomic status is critically assessed based on a study of type specimens and new material from Canada and the United States. The following 3 species, supposedly strictly specific to their fish definitive hosts, are recognized as valid: (1) Proteocephalus fluviatilis Bangham, 1925 (new synonyms Proteocephalus osburni Bangham, 1925 and Proteocephalus microcephalus Haderlie, 1953 ; Proteocephalus 'robustus' nomen nudum) from the smallmouth and largemouth bass, Micropterus dolomieu (Lacépède) (type host) and Micropterus salmoides (Lacépède) (both Centrarchidae); (2) Proteocephalus luciopercae Wardle, 1932 (new synonym Proteocephalus stizostethi Hunter and Bangham, 1933 ) from the walleye, Sander vitreus (Mitchill) (type host), and sauger, Sander canadensis (Griffith et Smith) (Percidae); and (3) Proteocephalus pearsei La Rue, 1919 , a parasite of the yellow perch, Perca flavescens Mitchill (Percidae). All species are illustrated based on new, properly heat-fixed material. Scanning electron micrographs of the scoleces of percid tapeworms P. luciopercae and P. pearsei, as well as the bass tapeworms P. fluviatilis and Proteocephalus ambloplitis ( Leidy, 1887 ), the latter of which does not belong to this Proteocephalus-aggregate, are provided for the first time together with a simple key to species identification of proteocephalids from centrarchiform and perciform teleost fishes.

Bluegill Picornavirus isolated from a mortality event involving Bluegill (Lepomis macrochirus) in the upper Mississippi River.

A mortality event involving an estimated 1,000 adult bluegills (Lepomis macrochirus) was observed in an ice-covered backwater lake of the upper Mississippi River near Alma, Wisconsin, in December of 2017. Macroscopic signs of disease included abdominal distension due to fluid accumulation within the internal organs as well as external and internal haemorrhaging. Histological evaluation revealed chronic peritonitis with peritoneal adhesions in all fish examined. Kidney, spleen and ascites fluid samples were collected from diseased bluegills and examined for the presence of pathogens. Bluegill picornavirus (BGPV) was isolated using tissue cell culture methods utilizing a recently developed, uncharacterized bluegill fry cell line (BF-4), and the presence of this virus was confirmed through molecular identification. The current geographic range, known susceptible hosts as well as historical epizootics associated with BPGV is discussed. The ability of BGPV to cause significant mortality in wild fish further emphasizes the importance of monitoring both wild and hatchery populations for this pathogen.

Non-native parasitic copepod Neoergasilus japonicus (Harada, 1930) utilizes non-native fish host Lepomis gibbosus (L.) in the floodplain of the River Dyje (Danube basin).

The parasitic copepod Neoergasilus japonicus (Harada, 1930) (Ergasilidae), native to east Asia, is widely distributed in Asia, Europe, and North and Central America. Recently, this species appeared in lentic water bodies of the River Dyje floodplain (Danube basin, Czech Republic). It was first recorded in 2015 and in 2 years it reached a 100% prevalence in recently expanding non-native fish host, Lepomis gibbosus (Linnaeus, 1758) (Centrarchidae, native to North America) at two borrow pits. Abundance of N. japonicus increased with fish length, with maximum intensity of infection reaching 99 parasites per fish. The parasite was most frequently found attached to the dorsal and anal fins of fish, while preference for the dorsal fin was more evident with lower infection intensities. Utilization of expanding fish hosts in water bodies that are regularly interconnected via natural or managed flooding may support the rapid dispersal of this non-native parasite.

Changing environmental gradients over forty years alter ecomorphological variation in Guadalupe Bass Micropterus treculii throughout a river basin.

Understanding the degree of intraspecific variation within and among populations is a key aspect of predicting the capacity of a species to respond to anthropogenic disturbances. However, intraspecific variation is usually assessed at either limited temporal, but broad spatial scales or vice versa, which can make assessing changes in response to long-term disturbances challenging. We evaluated the relationship between the longitudinal gradient of changing flow regimes and land use/land cover patterns since 1980 and morphological variation of Guadalupe Bass Micropterus treculii throughout the Colorado River Basin of central Texas. The Colorado River Basin in Texas has experienced major alterations to the hydrologic regime due to changing land- and water-use patterns. Historical collections of Guadalupe Bass prior to rapid human-induced change present the unique opportunity to study the response of populations to varying environmental conditions through space and time. Morphological differentiation of Guadalupe Bass associated with temporal changes in flow regimes and land use/land cover patterns suggests that they are exhibiting intraspecific trait variability, with contemporary individuals showing increased body depth, in response to environmental alteration through time (specifically related to an increase in herbaceous land cover, maximum flows, and the number of low pulses and high pulses). Additionally, individuals from tributaries with increased hydrologic alteration associated with urbanization or agricultural withdrawals tended to have a greater distance between the anal and caudal fin. These results reveal trait variation that may help to buffer populations under conditions of increased urbanization and sprawl, human population growth, and climate risk, all of which impose novel selective pressures, especially on endemic species like Guadalupe Bass. Our results contribute an understanding of the adaptability and capacity of an endemic population to respond to expected future changes based on demographic or climatic projection.

Correlates of elemental-isotopic composition of stream fishes: the importance of land-use, species identity and body size.

The isotopic (δ13 C and δ15 N) and stoichiometric (C:N:P) compositions of four fish species (Family Centrarchidae: Lepomis auritus, Lepomis cyanellus; Family Cyprinidae: Nocomis leptocephalus, Semotilus atromaculatus) were examined across four North Carolina Piedmont streams arrayed along an urbanization gradient. Both isotopic and stoichiometric composition of fishes appeared to track changes occurring in basal resource availability. Values of δ13 C of basal resources and consumers were more enriched at the most urbanized streams. Similarly, basal resources and consumers were δ15 N-enriched at more urbanized streams. Basal resource stoichiometry varied across streams, with periphyton being the most variable. Primary consumers stoichiometry also differed across streams. Intraspecific variation in fish stoichiometry correlated with the degree of urbanization, as the two cyprinids had higher N content and L. cyanellus had higher P content in more urbanized streams, probably due to enrichment of basal resources. Intrinsic factors, specifically species identity and body size also affected stoichiometric variation. Phosphorus (P) content increased significantly with body size in centrarchids, but not in cyprinids. These results suggest that although species identity and body size are important predictors of elemental stoichiometry, the complex nature of altered urban streams may yield imbalances in the elemental composition of consumers via their food resources.

Life history constraints explain negative relationship between fish productivity and dissolved organic carbon in lakes.

Resource availability constrains the life history strategies available to organisms and may thereby limit population growth rates and productivity. We used this conceptual framework to explore the mechanisms driving recently reported negative relationships between fish productivity and dissolved organic carbon (DOC) concentrations in lakes. We studied populations of bluegill (Lepomis macrochirus) in a set of lakes with DOC concentrations ranging from 3 to 24 mg/L; previous work has demonstrated that primary and secondary productivity of food webs is negatively related to DOC concentration across this gradient. For each population, we quantified individual growth rate, age at maturity, age-specific fecundity, maximum age, length-weight and length-egg size relationships, and other life history characteristics. We observed a strong negative relationship between maximum size and DOC concentration; for instance, fish reached masses of 150 to 260 g in low-DOC lakes but <120 g in high-DOC lakes. Relationships between fecundity and length, and between egg size and length, were constant across the DOC gradient. Because fish in high-DOC lakes reached smaller sizes but had similar fecundity and egg size at a given size, their total lifetime fecundity was as much as two orders of magnitude lower than fish in low-DOC lakes. High DOC concentrations appeared to constrain the range of bluegill life history strategies available; populations in high-DOC lakes always had low initial growth rates and high ages at maturity, whereas populations in low-DOC showed higher variability in these traits. This was also the case for the intrinsic rates of natural increase of these populations, which were always low at the high end of the DOC gradient. The potentially lower capacity for fish populations in high-DOC lakes to recover from exploitation has clear implications for the sustainable management of recreational fisheries in the face of considerable spatial heterogeneity and ongoing temporal change in lake DOC concentrations.

Biodiversity data mining from Argus-eyed citizens: the first illegal introduction record of Lepomis macrochirus macrochirus Rafinesque, 1819 in Japan based on Twitter information.

An apparent illegal introduction of Lepomis macrochirus macrochirus from Yokohama City, Kanagawa Prefecture, Japan, is reported based on a juvenile specimen and a photograph of two adults collected on 14 June 2015 and deposited in the Kangawa Prefectural Museum of Natural History. The specimens and photographs were initially reported on the internet-based social networking site, Twitter. Two specimens of Carassius auratus, including an aquarium form, were also reported at the same locality and date, suggesting that the illegal introductions originated from an aquarium release. Our report demonstrates an example of web data mining in the discipline of Citizen Science.

New species of Proterometra (Digenea: Azygiidae) and its life cycle in the Chickasawhay River, Mississippi, USA, with supplemental observations of Proterometra autraini.

We describe Proterometra ariasae n. sp. based upon cercariae shed from a freshwater snail, Pleurocera sp., and adults infecting the buccal cavity of longear sunfish, Lepomis megalotis, captured from the Chickasawhay River, Mississippi, USA. We also provide supplemental observations of cercarial and adult paratypes of Proterometra autraini from the Au Train River, Michigan, USA. Sequence data for the ribosomal internal transcribed spacer 2 (ITS2) from adults and cercariae of the new species were identical. Adults of the new species differ from congeners by having (i) a markedly large body, (ii) a proportionally large oral sucker, (iii) ovoid testes, (iv) a strongly muscular and laterally expanded pars prostatica, (v) a uterus that is extensively convoluted between the ovary and ventral sucker (vi) and a vitellarium as long as the caeca and extending posteriad beyond the anterior margin of the testes. Cercariae of the new species differ from those of its congeners by having (i) a tail stem that is shorter than 10mm and that lacks a medial constriction, (ii) obcordate furcae that are wider than long, (iii) mamillae distributed throughout the anterior tail stem only, and (v) a proportionally small distome that has relatively few uterine eggs and remains withdrawn in the anterior tail stem region in actively swimming cercariae. This is the first report of Proterometra from Mississippi, the second description to employ morphology and sequence data to elucidate a life cycle for Proterometra, and the third species of Proterometra from an intermediate host not assigned to Elimia.

Developing major histocompatibility markers in a species of concern: the Sacramento perch Archoplites interruptus.

Primers targeting two non-neutral major histocompatibility complex (mhc) II ß genes were developed and assayed across several disjoint Sacramento perch Archoplites interruptus sampling locations. Variability at the two mhc loci among sampling stocks strongly correlated to previous estimates with neutral markers, suggesting that the effect of genetic drift was not limited to neutrally evolving regions of the genome. The novel mhc primers will help develop admixture schemes in A. interruptus captive breeding programmes and will increase the success of future reintroductions of this species of concern.

Interspecific relationships and the evolution of sexual dimorphism in pygmy sunfishes (Centrarchidae: Elassoma).

The genus Elassoma represents a small but unique component of the aquatic biodiversity hotspot in southeastern North America. We present the first phylogeny of the seven described species, corroborated by sequence data from mitochondrial and nuclear protein coding genes. This analysis reveals a Coastal Plain clade sister to the geographically isolated, and federally protected, Elassoma alabamae. The Coastal Plain clade contains the widespread E. zonatum, which is sister to a clade primarily restricted to lowland Neogene subprovinces. We analyzed morphometric data in a phylogenetic context to illustrate the evolution of sexual shape dimorphism within the genus. Sixteen univariate and three multivariate traits were tested for significant sexual dimorphism for each species, and relative transformation rates were inferred from the time tree. A simple index of interspecific sexual dimorphism revealed greater disparity among sympatric species comparisons than among allopatric comparisons. Results implicate geology as a primary factor influencing ecological diversification, and sexual selection as a mechanism reinforcing reproductive isolation in areas of secondary contact. We discuss putative roles of geological history and sexual selection in the generation and maintenance of the aquatic biodiversity gradient in southeastern North America.

Ancestral character estimation under the threshold model from quantitative genetics.

Evolutionary biology is a study of life's history on Earth. In researching this history, biologists are often interested in attempting to reconstruct phenotypes for the long extinct ancestors of living species. Various methods have been developed to do this on a phylogeny from the data for extant taxa. In the present article, I introduce a new approach for ancestral character estimation for discretely valued traits. This approach is based on the threshold model from evolutionary quantitative genetics. Under the threshold model, the value exhibited by an individual or species for a discrete character is determined by an underlying, unobserved continuous trait called "liability." In this new method for ancestral state reconstruction, I use Bayesian Markov chain Monte Carlo (MCMC) to sample the liabilities of ancestral and tip species, and the relative positions of two or more thresholds, from their joint posterior probability distribution. Using data simulated under the model, I find that the method has very good performance in ancestral character estimation. Use of the threshold model for ancestral state reconstruction relies on a priori specification of the order of the discrete character states along the liability axis. I test the use of a Bayesian MCMC information theoretic criterion based approach to choose among different hypothesized orderings for the discrete character. Finally, I apply the method to the evolution of feeding mode in centrarchid fishes.