Status of the Federally Endangered Alabama Cavefish, Speoplatyrhinus Poulsoni (Amblyopsidae), in Key Cave and surrounding Caves, Alabama

Speoplatyrhinus poulsoni, the Alabama cavefish, is a federally endangered stygobitic fish that is restricted to Key Cave in northwestern Alabama. Due to the low abundance of this species and the continuous threats to the cave's recharge area, we surveyed Key Cave from 1992 to 1997. Alabama cavefish were present in five different pools and had numbers comparable to previous surveys performed in the 1970's and 1980's. Three different size classes were observed, indicating recruitment is occurring. These data suggest that the population is relatively stable, but future monitoring is recommended. The Alabama cavefish is an endangered species and is extremely difficult to capture, therefore any marking program to accurately estimate the population size is unlikely. A single specimen of Typhlichthys subterraneus was captured in Key Cave. This falsifies the previous assumption that these two species are allopatric. Because of this discovery, we resurveyed seven caves in the area around Key Cave for additional populations of S. poulsoni. These caves had either known populations of T. subterraneus or unidentified cavefish. No additional populations of S. poulsoni were discovered, but further investigation is warranted. The Key Cave National Wildlife Refuge was established in the high recharge area of Key Cave to protect this habitat, yet threats to the groundwater continue from encroaching urbanization.     

Quantitative Lipidomics and Spatial MS-Imaging Uncovered Neurological and Systemic Lipid Metabolic Pathways Underlying Troglomorphic Adaptations in Cave-Dwelling Fish

Sinocyclocheilus represents a rare, freshwater teleost genus endemic to China that comprises the river-dwelling surface fish and the cave-dwelling cavefish. Using a combinatorial approach of quantitative lipidomics and mass-spectrometry imaging (MSI), we demonstrated that neural compartmentalization of lipid distribution and lipid metabolism is associated with the evolution of troglomorphic traits in Sinocyclocheilus. Attenuated docosahexaenoic acid (DHA) biosynthesis via the Δ4 desaturase pathway led to reductions in DHA-phospholipids in cavefish cerebellum. Instead, cavefish accumulates arachidonic acid-phospholipids that may disfavor retinotectal arbor growth. Importantly, MSI of sulfatides coupled with immunostaining of myelin basic protein and transmission electron microscopy images of hindbrain axons revealed demyelination in cavefish raphe serotonergic neurons. Demyelination in cavefish parallels the loss of neuroplasticity governing social behavior such as aggressive dominance. Outside the brain, quantitative lipidomics and qRT-PCR revealed systemic reductions in membrane esterified DHAs in the liver, attributed to suppression of genes along the Sprecher pathway (elovl2, elovl5, and acox1). Development of fatty livers was observed in cavefish; likely mediated by an impeded mobilization of storage lipids, as evident in the diminished expressions of pnpla2, lipea, lipeb, dagla, and mgll; and suppressed β-oxidation of fatty acyls via both mitochondria and peroxisomes as reflected in the reduced expressions of cpt1ab, hadhaa, cpt2, decr1, and acox1. These neurological and systemic metabolic adaptations serve to reduce energy expenditure, forming the basis of recessive evolution that eliminates nonessential morphological and behavioral traits and giving cavefish a selective advantage to thrive in caves where proper resource allocation becomes a major determinant of survival.     

Length–weight relationships of two cavefish species,Sinocyclocheilus jii Zhang & Dai, 1992 and Sinocyclocheilus guilinensis Ji, 1985 (Cypriniformes: Cyprinidae)

Length–weight relationships of two cavefish species, Sinocylocheilus jii Zhang & Dai, (Acta Zootaxonomica Sinica, 17, 1992, 377) and Sinocyclocheilus guilinensis Ji, 1985, were calculated based on data from 186 specimens sampled by gill nets and fish traps from subterranean rivers of the Pearl River basin in China between 2008 and 2014. Results indicate that mean LWR parameters b for these two cavefish species are 2.88 and 2.61, respectively. A maximum size was recorded for S. jii.     

Complex Evolutionary and Genetic Patterns Characterize the Loss of Scleral Ossification in the Blind Cavefish Astyanax mexicanus

The sclera is the tough outer covering of the eye that provides structural support and helps maintain intraocular pressure. In some fishes, reptiles, and birds, the sclera is reinforced with an additional ring of hyaline cartilage or bone that forms from scleral ossicles. Currently, the evolutionary and genetic basis of scleral ossification is poorly understood, especially in teleost fishes. We assessed scleral ossification among several groups of the Mexican tetra (Astyanax mexicanus), which exhibit both an eyed and eyeless morph. Although eyed Astyanax surface fish have bony sclera similar to other teleosts, the ossicles of blind Astyanax cavefish generally do not form. We first sampled cavefish from multiple independent populations and used ancestral character state reconstructions to determine how many times scleral ossification has been lost. We then confirmed these results by assessing complementation of scleral ossification among the F₁ hybrid progeny of two cavefish populations. Finally, we quantified the number of scleral ossicles present among the F₂ hybrid progeny of a cross between surface fish and cavefish, and used this information to identify quantitative trait loci (QTL) responsible for this trait. Our results indicate that the loss of scleral ossification is common–but not ubiquitous–among Astyanax cavefish, and that this trait has been convergently lost at least three times. The presence of wild-type, ossified sclera among the F₁ hybrid progeny of a cross between different cavefish populations confirms the convergent evolution of this trait. However, a strongly skewed distribution of scleral ossicles found among surface fish x cavefish F₂ hybrids suggests that scleral ossification is a threshold trait with a complex genetic basis. Quantitative genetic mapping identified a single QTL for scleral ossification on Astyanax linkage group 1. We estimate that the threshold for this trait is likely determined by at least three genetic factors which may control the severity and onset of lens degeneration in cavefishes. We conclude that complex evolutionary and genetic patterns underlie the loss of scleral ossification in Astyanax cavefish.     

Two new species of Melanoleuca (Agaricales,Basidiomycota) from northeastern China,supported by morphological and molecular data

Two new species, Melanoleuca leucopoda and M. porphyropoda, are described based on collections made from Shenyang City, Liaoning Province, China. Melanoleuca leucopoda is mainly characterized by its whitish stipe with fibrils and oblong spores with elongated warts. Melanoleuca porphyropoda differs from all other Melanoleuca species in lacking cystidia and in having decurrent gills and a purplish stipe. The sequences of internal transcribed spacer regions (ITS1-5.8S-ITS2) of Melanoleuca species were analyzed and the results indicated that two new species clustered into two clades and differed from the other species of the genus. The combination of morphological and molecular data confirmed that the two fungi are new species. The morphological similarity of the new species to other species of Melanoleuca and the systematic position of the two species based on molecular data are also discussed.     

Albinism in phylogenetically and geographically distinct populations of Astyanax cavefish arises through the same loss-of-function Oca2 allele

The Mexican tetra, Astyanax mexicanus, comprises 29 populations of cave-adapted fish distributed across a vast karst region in northeastern Mexico. These populations have a complex evolutionary history, having descended from ‘old'' and ‘young'' ancestral surface-dwelling stocks that invaded the region ∼6.7 and ∼2.8 MYa, respectively. This study investigates a set of captive, pigmented Astyanax cavefish collected from the Micos cave locality in 1970, in which albinism appeared over the past two decades. We combined novel coloration analyses, coding sequence comparisons and mRNA expression level studies to investigate the origin of albinism in captive-bred Micos cavefish. We discovered that albino Micos cavefish harbor two copies of a loss-of-function ocular and cutaneous albinism type II (Oca2) allele previously identified in the geographically distant Pachón cave population. This result suggests that phylogenetically young Micos cavefish and phylogenetically old Pachón cave fish inherited this Oca2 allele from the ancestral surface-dwelling taxon. This likely resulted from the presence of the loss-of-function Oca2 haplotype in the ‘young'' ancestral surface-dwelling stock that colonized the Micos cave and also introgressed into the ancient Pachón cave population. The appearance of albinism in captive Micos cavefish, caused by the same loss-of-function allele present in Pachón cavefish, implies that geographically and phylogenetically distinct cave populations can evolve the same troglomorphic phenotype from standing genetic variation present in the ancestral taxon.     

To see or not to see: evolution of eye degeneration in mexican blind cavefish

The evolutionary mechanisms responsible for the loss of eyesin cave animals are still unresolved. Hypotheses invoking naturalselection or neutral mutation have been advanced to explaineye regression. Here we describe comparative molecular and developmentalstudies in the teleost Astyanax mexicanus that shed new lighton this problem. A. mexicanus is a single species consistingof a sighted surface-dwelling form (surface fish) and many blindcave-dwelling forms (cavefish) from different caves. We firstreview the evolutionary relationships of Astyanax cavefish populationsand conclude that eye degeneration may have evolved multipletimes. We then compare the mechanisms of eye degeneration indifferent cavefish populations. We describe the results of experimentsshowing that programmed cell death of the lens plays a key rolein controlling eye degeneration in these cavefish populations.We also show that Pax6 gene expression and fate determinationin the optic primordia are modified similarly in different cavefishpopulations, probably due to hyperactive midline signaling.We discuss the contributions of the comparative developmentalapproach toward resolving the evolutionary mechanisms of eyedegeneration. A new hypothesis is presented in which both naturalselection and neutral mutation are proposed to have roles incavefish eye degeneration.     

A Potential Benefit of Albinism in Astyanax Cavefish: Downregulation of the oca2 Gene Increases Tyrosine and Catecholamine Levels as an Alternative to Melanin Synthesis

Albinism, the loss of melanin pigmentation, has evolved in a diverse variety of cave animals but the responsible evolutionary mechanisms are unknown. In Astyanax mexicanus, which has a pigmented surface dwelling form (surface fish) and several albino cave-dwelling forms (cavefish), albinism is caused by loss of function mutations in the oca2 gene, which operates during the first step of the melanin synthesis pathway. In addition to albinism, cavefish have evolved differences in behavior, including feeding and sleep, which are under the control of the catecholamine system. The catecholamine and melanin synthesis pathways diverge after beginning with the same substrate, L-tyrosine. Here we describe a novel relationship between the catecholamine and melanin synthesis pathways in Astyanax. Our results show significant increases in L-tyrosine, dopamine, and norepinephrine in pre-feeding larvae and adult brains of Pachón cavefish relative to surface fish. In addition, norepinephrine is elevated in cavefish adult kidneys, which contain the teleost homologs of catecholamine synthesizing adrenal cells. We further show that the oca2 gene is expressed during surface fish development but is downregulated in cavefish embryos. A key finding is that knockdown of oca2 expression in surface fish embryos delays the development of pigmented melanophores and simultaneously increases L-tyrosine and dopamine. We conclude that a potential evolutionary benefit of albinism in Astyanax cavefish may be to provide surplus L-tyrosine as a precursor for the elevated catecholamine synthesis pathway, which could be important for adaptation to the challenging cave environment.     

Population Biology and Growth of Ozark Cavefish in Logan Cave National Wildlife Refuge, Arkansas

Ozark cavefish, Amblyopsis rosae, is a threatened species endemic to the Springfield Plateau of the Ozark Highlands in Arkansas, Missouri, and Oklahoma. One of the largest known Ozark cavefish populations, located in Logan Cave, Arkansas, was surveyed 25 times over a two-year period between 1993 and 1995. During the study, 147 Ozark cavefish > 30 mm (TL) were marked with visual implant tags and 140 Ozark cavefish were available for recapture; 68 were recaptured 189 times and the rest (72) were never recaptured. Individual Ozark cavefish persisted in Logan Cave for a relatively short time. Only 14% of 80 fish tagged during a previous study in 1992 were recaptured during this study, and half of all recaptured fish disappeared within three months. However, if a fish persisted for at least seven months in the cave, its probability of being recaptured over an additional year was high. Maximum persistence of a tagged fish was 28 months, suggesting these fish have a maximum life-span of 4–5 years. Growth averaged 0.6 mm per month, with maximum recorded growth of 6 mm per month and a maximum size of 65 mm TL. Smaller fish grew faster than larger fish but growth rates were sporadic, with several mid-sized fish (45–49 mm) showing little growth (0–3 mm year^-1) while some fish > 50 mm grew up to 12 mm year^-1. Most fish gained in length during April–October, the same period a maternity colony of gray bats occupied the cave. Gross Ozark cavefish movement over the study period ranged up to 1002 m, with a mean movement of 1.2 m day^-1; movement was positively correlated with Ozark cavefish total length. Death seemed the most likely explanation for loss of tagged Ozark cavefish, including fish that emigrated out of the cave. Little up-stream movement was recorded between reaches and did not account for loss of tagged fish. Reproduction within the cave and immigration from the aquifer accounted for persistence of Ozark cavefish in Logan Cave.     

The description of a new species of the Neotropical land crab genus Gecarcinus Leach, 1814 (Crustacea,Decapoda, Brachyura,Gecarcinidae)

In this contribution a new species of the land crab genus Gecarcinus Leach, 1814, from the Neotropical Pacific coast of South America is described and illustrated. In addition to its unique body color, Gecarcinus nobilii sp. n. is distinguished from congeners by a distinctly wider carapace front and differences in the shape of the infraorbital margin. The new species is not isolated from Gecarcinus populations from the Pacific coast of Central America by an insurmountable geographic barrier. Considering the closure of the Panamanian Isthmus as a calibration point for morphological divergence between the trans-isthmian mainland populations of Gecarcinus, the virtual lack of morphological differentiation (other than color) between them and the distinctness of G. nobilii sp. n. suggests that G. nobilii sp. n. evolved from a common ancestor before the Isthmus closed.     

Metapochonia hahajimaensis (Clavicipitaceae,Hypocreales), a new species from soil in Hahajima Island,Tokyo, Japan

A new species, Metapochonia hahajimaensis is proposed. Three isolates were obtained from soil samples collected from Hahajima, Ogasawara Islands, Tokyo, Japan. Using a combination of morphological characteristics and multigene (ITS, SSU, LSU, TEF) phylogenics, the isolates were identified as a Metapochonia species which were previously undescribed. The new species is distinguished from the closely-related M. bulbillosa and M. goniodes or other Metapochonia taxa by conidial size and colonial characteristics.     

Brachylaima succini sp. nov. (Trematoda: Brachylaimidae) from Succinea lauta,an amber snail in Hokkaido,Japan

Through a continuous survey of trematodes in land snails of Hokkaido, the northernmost island of Japan, we have discovered four species of the genus Brachylaima (Trematode: Brachylaimidae). Among them, Brachylaima ezohelicis, Brachylaima asakawai, and Brachylaima lignieuhadrae have already been described. Each of the three species is a strict specialist in selecting a particular species of land snail as the first intermediate host. In this report, we propose the fourth species, Brachylaima succini sp. nov., based on ecological, morphological, and phylogenetic considerations. Sporocysts and metacercariae of the new species were found exclusively from Succinea lauta, which is known as an amber snail indigenous to Hokkaido. Phylogenetic trees of nuclear 28S rDNA and mitochondrial cytochrome c oxidase subunit 1 (cox1) demonstrated it to be distinct from the other sympatric species. Although metacercariae of the new species possessed unique morphological characters, adult worms experimentally raised from the metacercariae were similar to those of B. ezohelicis and B. lignieuhadrae. Natural definitive hosts of the new species are unknown, but the existence of common cox1 haplotypes from far-distant localities suggests a possibility that birds are involved as the definitive hosts. Findings of amber snails coinfected with both sporocysts of the new species and Leucochloridium perturbatum also support the involvement of birds.     

Morphological and phylogenetic studies of Cytospora (Valsaceae,Diaporthales) isolates from Chinese scholar tree,with description of a new species

Cytospora species are the most serious and widespread pathogens associated with canker disease on multiple plants. In this study, three species, i.e., Cytospora sophoricola, C. chrysosperma, and C. sophorae, which were isolated from Sophora in China, are described and illustrated based on their morphological characteristics and phylogenetic analyses. Cytospora sophoricola was distinguished clearly by its larger disc, multiple ostioles, cystic and multiple locules, and specific cultural characteristics, i.e., protruding fruiting bodies. Maximum parsimony and maximum likelihood analysis showed that it did not cluster with any known species of Cytospora, so it is described as a new species. Cytospora sophorae is a previously reported species from Sophora, which is redescribed based on new isolates and additional observations. Another species was identified as C. chrysosperma, which is reported for the first time on Sophora, so Papilionaceae is shown to be a new host family for C. chrysosperma. The morphological affinities of these species with related taxa are discussed, while the phylogenetic relationships of these species with other fungus in the genus Cytospora were elucidated based on their internal transcribed spacer (ITS) rRNA region sequences.     

A New Species of Muscicapa Flycatcher from Sulawesi,Indonesia

The Indonesian island of Sulawesi, a globally important hotspot of avian endemism, has been relatively poorly studied ornithologically, to the extent that several new bird species from the region have been described to science only recently, and others have been observed and photographed, but never before collected or named to science. One of these is a new species of Muscicapa flycatcher that has been observed on several occasions since 1997. We collected two specimens in Central Sulawesi in 2012, and based on a combination of morphological, vocal and genetic characters, we describe the new species herein, more than 15 years after the first observations. The new species is superficially similar to the highly migratory, boreal-breeding Gray-streaked Flycatcher Muscicapa griseisticta, which winters in Sulawesi; however, the new species differs strongly from M. griseisticta in several morphological characters, song, and mtDNA. Based on mtDNA, the new species is only distantly related to M. griseisticta, instead being a member of the M. dauurica clade. The new species is evidently widely distributed in lowland and submontane forest throughout Sulawesi. This wide distribution coupled with the species'' apparent tolerance of disturbed habitats suggests it is not currently threatened with extinction.     

Two new species of gorgonian octocorals from the Tropical Eastern Pacific Biogeographic Region (Cnidaria,Anthozoa, Gorgoniidae)

The gorgoniid Eugorgia is exclusively an eastern Pacific genus. It has a wide geographic and bathymetric range of distribution, found from California to Perú and extends down to 65 m deep. Two new species are herein described. The morphological characters were analyzed and illustrated by light and scanning electron microscopy. Eugorgia beebei sp. n. can be distinguished by its white, ascending, sparse colony growth. Eugorgia mutabilis sp. n. can be distinguished by its white colony that changes color after collection, and the conspicuous sharp-crested disc sclerites. From a morphological point of view the new species are related to the daniana-group, the rubens-group and the siedenburgae-group of Eugorgia; their affiliations, and the proposal of a new group are discussed. These new species increases the number of species in the genus to 15, and contribute to the knowledge of the eastern Pacific octocoral biodiversity.     

Deep phylogenetic divergence between Scolytoplatypus and Remansus,a new genus of Scolytoplatypodini from Madagascar (Coleoptera,Curculionidae, Scolytinae)

Scolytoplatypodini Blandford is a monotypic tribe of ambrosia beetles found in Asia, Madagascar and Africa. Only three species are currently known from Madagascar and four additional species are here described as new to science. Phylogenetic analyses of morphological and molecular data revealed that four of the seven endemic species are deeply separated from all other species by genetic and distinct morphological characters and therefore placed in a new genus Remansus Jordal. The split between this ancient lineage and Scolytoplatypus Schaufuss was estimated to approximate Palaeocene age (63 Ma), extending the minimum age of ambrosia feeding for this tribe to the beginning of the Palaeocene‒Eocene thermal maximum (PETM). In addition to the ancient origin of Remansus in Madagascar during the Palaeocene, a second origin occurred in Scolytoplatypus no more than 13 Ma. A geographical origin of the latter in South-Eastern Africa was unequivocally inferred from the phylogenies.     

Description of a new species of Parasesarma (Crustacea; Decapoda; Brachyura; Sesarmidae) from the Persian Gulf, based on morphological and genetic characteristics

The genera Parasesarma and Perisesarma are common inhabitants of mangroves and estuaries of the Indo-West Pacific. These, overall similar, genera of sesarmid crabs can be told apart by the presence or absence of an anterolateral tooth, but recent and ongoing studies show that they are not reciprocally monophyletic. In recent years, detailed morphological studies have revealed a surprising number of undescribed new species within these genera. In the present paper, we describe a new species of the genus Parasesarma from the relatively poorly studied Persian Gulf. So far, these specimens had been referred to as Parasesarma plicatum (Latreille, 1803), but morphological and genetic characteristics clearly allow separation of the two species. Consequently, the species from the Persian Gulf is here described as new and morphological and mitochondrial genetic characters are provided that allow recognition of the species.