Mycosporine-like amino acid content in four species of sea anemones in the genus Anthopleura reflects phylogenetic but not environmental or symbiotic relationships

We examine the occurrence of UV-absorbing, mycosporine-like amino acids (MAAs) in four sympatric species of sea anemones in the genus Anthopleura, all collected from intertidal habitats on the Pacific Coast of temperate North America. We compare patterns of MAAs in A. elegantissima of several types: specimens having predominately zooxanthellae (dinoflagellates comprising at least two species) or zoochlorellae as symbionts; those containing algal endosymbionts of both kinds, and naturally occurring aposymbiotic specimens that lack the endosymbionts typically found in most specimens. We also compare MAAs in zooxanthellate specimens of A. sola and A. xanthogrammica, and specimens from the asymbiotic species A. artemisia. Our findings indicate that the complements of the four major MAAs in these species of Anthopleura (mycosporine-taurine, shinorine, porphyra-334, and mycosporine-2 glycine) broadly reflect phylogenetic differences among the anemones rather than the taxon of endosymbionts, presence or absence of symbionts, or environmental factors. An exception, however, occurs in A. elegantissima, where mycosporine-2 glycine increases in concentration with the density of zooxanthellae. Our evidence also shows that A. elegantissima can accumulate MAAs from its food, which may explain the occasional occurrence of minor MAAs in some individuals.     

Sources of mycosporine-like amino acids in planktonic Chlorella-bearing ciliates (Ciliophora)

1. Mycosporine‐like amino acids (MAAs) are a family of secondary metabolites known to protect organisms exposed to solar UV radiation. We tested their distribution among several planktonic ciliates bearing Chlorella isolated from an oligo‐mesotrophic lake in Tyrol, Austria. In order to test the origin of these compounds, the MAAs were assessed by high performance liquid chromatography in both the ciliates and their symbiotic algae. 2. Considering all Chlorella‐bearing ciliates, we found: (i) seven different MAAs (mycosporine‐glycine, palythine, asterina‐330, shinorine, porphyra‐334, usujirene, palythene); (ii) one to several MAAs per species and (iii) qualitative and quantitative seasonal changes in the MAAs (e.g. in Pelagodileptus trachelioides). In all species tested, concentrations of MAAs were always <1% of ciliate dry weight. 3. Several MAAs were also identified in the Chlorella isolated from the ciliates, thus providing initial evidence for their symbiotic origin. In Uroleptus sp., however, we found evidence for a dietary source of MAAs. 4. Our results suggest that accumulation of MAAs in Chlorella‐bearing ciliates represents an additional benefit of this symbiosis and an adaptation for survival in sunlit, UV‐exposed waters.     

Leptobrachium bompu(Amphibia,Anura, Megophryidae) Discovered in Upper Medog,Tibet, China with Descriptions of its Tadpoles,Advertisement Calls and Systematic Position

Leptobrachium bompu Sondhi and Ohler, 2011 was described based on a single specimen collected from Eaglenest in southern slope of Himalaya(holotype: No. KA0001/200905). In April, 2014, seven adults and three tadpoles of Leptobrachium bompu were collected from upper Medog, Tibet, China since most the morphological characters of the adult frogs matched the species Leptobrachium bompu, while the tadpoles mentioned above were determined as the same species as the adults by molecular systematics analysis. Three types of iris coloration were found in the seven adult specimens and two types of spiraculum in the three tadpoles. Advertisement calls usually had 6-8 notes and the fundamental frequency ranged from 1076 to 1466 Hz. All the specimens collected at upper Medog were clustered as one lineage with very low genetic variation and located at the base of the phylogenetic tree of the genus Leptobrachium.     

Chemical profiling of mycosporine‐like amino acids in twenty‐three red algal species

Rhodophyta produce a variety of chemically different mycosporine‐like amino acids (MAAs), compounds that are known as some of the strongest ultraviolet (UV) absorbing molecules in nature. Accordingly, they primarily act as photoprotectants against harmful levels of solar ultraviolet radiation in the UV‐A and UV‐B range. In order to get a deeper understanding of the chemical diversity of MAAs in red algae, pure standards of eleven mycosporine‐like amino acids were isolated from three different species (Agarophyton chilense, Pyropia plicata and Champia novae‐zelandiae) using various chromatographic methods. Their structures were confirmed by nuclear magnetic resonance and mass spectrometry. Four out of the eleven MAAs are reported for the first time in algae. In addition, a new high‐performance liquid chromatography method was developed for the separation of all isolated MAAs and successfully applied for the analysis of twenty‐three red algal species of marine origin. All of them contained MAAs, the most abundant compounds were shinorine, palythine, asterina‐330 and porphyra‐334. For some samples, the direct assignment of MAAs based on their UV spectra was not possible; therefore, the target analytes were enriched by a simple concentration step, followed by liquid chromatography‐mass spectrometry analysis of the extracts. This approach enabled a deeper insight into the MAA pattern of red algae, indicating that not only the four dominant ones are synthesized but also many others, which were often described as unknown compounds in previous studies.     

Are geometric morphometric analyses replicable? Evaluating landmark measurement error and its impact on extant and fossil Microtus classification

Geometric morphometric analyses are frequently employed to quantify biological shape and shape variation. Despite the popularity of this technique, quantification of measurement error in geometric morphometric datasets and its impact on statistical results is seldom assessed in the literature. Here, we evaluate error on 2D landmark coordinate configurations of the lower first molar of five North American Microtus (vole) species. We acquired data from the same specimens several times to quantify error from four data acquisition sources: specimen presentation, imaging devices, interobserver variation, and intraobserver variation. We then evaluated the impact of those errors on linear discriminant analysis‐based classifications of the five species using recent specimens of known species affinity and fossil specimens of unknown species affinity. Results indicate that data acquisition error can be substantial, sometimes explaining >30% of the total variation among datasets. Comparisons of datasets digitized by different individuals exhibit the greatest discrepancies in landmark precision, and comparison of datasets photographed from different presentation angles yields the greatest discrepancies in species classification results. All error sources impact statistical classification to some extent. For example, no two landmark dataset replicates exhibit the same predicted group memberships of recent or fossil specimens. Our findings emphasize the need to mitigate error as much as possible during geometric morphometric data collection. Though the impact of measurement error on statistical fidelity is likely analysis‐specific, we recommend that all geometric morphometric studies standardize specimen imaging equipment, specimen presentations (if analyses are 2D), and landmark digitizers to reduce error and subsequent analytical misinterpretations.     

The Good,the Bad,and the Ugly: The Influence of Skull Reconstructions and Intraspecific Variability in Studies of Cranial Morphometrics in Theropods and Basal Saurischians

Several studies investigating macroevolutionary skull shape variation in fossil reptiles were published recently, often using skull reconstructions taken from the scientific literature. However, this approach could be potentially problematic, because skull reconstructions might differ notably due to incompleteness and/or deformation of the material. Furthermore, the influence of intraspecific variation has usually not been explored in these studies. Both points could influence the results of morphometric analyses by affecting the relative position of species to each other within the morphospace. The aim of the current study is to investigate the variation in morphometric data between skull reconstructions based on the same specimen, and to compare the results to shape variation occurring in skull reconstructions based on different specimens of the same species (intraspecific variation) and skulls of closely related species (intraspecific variation). Based on the current results, shape variation of different skull reconstructions based on the same specimen seems to have generally little influence on the results of a geometric morphometric analysis, although it cannot be excluded that some erroneous reconstructions of poorly preserved specimens might cause problems occasionally. In contrast, for different specimens of the same species the variation is generally higher than between different reconstructions based on the same specimen. For closely related species, at least with similar ecological preferences in respect to the dietary spectrum, the degree of interspecific variation can overlap with that of intraspecific variation, most probably due to similar biomechanical constraints.     

UV‐Acclimation of Cultured Phytoplankton Exposed to Natural and Enhanced UVB Radiation and Methodological Aspects of MAA Quantification

Bloom‐forming dinoflagellates often synthesize high concentrations of mycosporine‐like amino acids (MAAs) as a protection from high PAR and UV radiation, which may afford them a competitive advantage over other species. Photoprotection and long‐term acclimation to natural and enhanced UVB radiation were examined during 4 weeks in three dinoflagellates and one diatom. Semi‐continuous, nutrient‐replete cultures were maintained outdoors under natural light, with or without additional UVB (from lamps). The photosynthetic performance (XE‐PAM fluorometry) and the concentration and composition of MAAs and photoprotective carotenoids (PPCs; comprising xanthophylls) were followed weekly. A great variety of MAAs was found in the dinoflagellates (up to 10, identified by cochromatography and LC–MS). Large differences between sp. (and between two strains of the same sp.) were observed. In most cases, the composition and concentration of MAAs and PPCs increased under enhanced UVB. Growth rate and photosynthetic performance were sometime lower under enhanced UVB, while in other cases photoacclimation seemed to occur. The least affected sp. was Alexandrium tamarense who responded most rapidly to enhanced UVB. The algae also responded to changes in natural incident radiation. Some methodological aspects of MAA quantification were also investigated (with four species of dinoflagellates). Our results indicate how freezing (liquid‐N) likely breaks the cells, mobilizing the water‐soluble MAAs in the filter moist (otherwise highly packaged in fresh cells), while the organelles containing the pigments remain relatively intact. The implications of standard sample handling on the evaluation of MAA concentration in algae and on the estimation of the absorption of suspended material in water from in vivo spectroscopy (filter pad technique) will also be discussed.     

Mycosporine-like amino acids protect the copepod Boeckella titicacae (Harding) against high levels of solar UVR

Studies were conducted in Lake Titicaca, Bolivia (16°S,69°W, 3810 m a.s.l.) to determine the effects of solar ultravioletradiation (UVR; 280–400 nm) and the protective role ofmycosporine-like amino acids (MAAs) in the calanoid copepodBoeckella titicacae (Harding). Zooplankton samples were takenfrom two different radiation environments: Lake Titicaca anda tank with running lake water, which was covered with layersof neutral-density screen for over a year, receiving ~10% ofincident radiation. Samples collected from the lake had significantlyhigher amounts of MAAs compared with those from the tank. Duringshort-term experiments (6–8 h incubations; six radiationtreatments using sharp cut-off Schott filters), significantlyhigher mortality was observed in B. titicacae from the tankthan from the lake. The higher sensitivity of B. titicacae containinglow amounts of MAAs (compared with specimens collected fromthe lake) was assessed through the determination of biologicalweighting functions (BWFs). A 3-day-long experiment performedwith specimens collected from the lake indicated an increasein mortality as a function of UVR exposure, reaching valuesas high as 70% at the end of day 3 for those individuals thatreceived full radiation compared with <10% for those thatreceived only photosynthetically active radiation (PAR; 400–700nm). Concomitantly, a reduction in the concentration of UV-absorbingcompounds as a function of UVR dose and decreasing wavelengthwas determined. The relatively high UVR dose threshold for mortality(~300 kJ m^–2), as well as a comparison with related species,demonstrated the high resistance of B. titicacae to UVR. Ourresults indicate that this resistance is provided by the presenceof protective compounds, i.e. MAAs, which were efficiently bioaccumulatedthrough the diet.     

Banded karyotypes of mole rats (Spalax, Spalacidae, Rodentia) from Turkey: a comparative analysis

Chromosome banding (G-, C- and Ag-NOR) analysis was carried out on 27 specimens of Sphalax ehrenbergi from seven localities and two specimens of S. leucodon from one locality, all from Turkey. No chromosomal variation was detected in S. ehrenbergi populations from Elazig, Siverek, Diyarbakir and Birecik having the same diploid numbers (2n = 52) and morphology of chromosomes (NFa = 72). The karyotypes of mole rats from Tarsus and Gaziantep possessed the identical diploid number (2n = 56) but different numbers of autosomal arms: NFa = 68 in the Tarsus and NFa = 78 in the Gaziantep populations. Chromosomes of S. leucodon from Malaty (2n = 60, NFa = 74) differed distinctly in the C-banding pattern from all S. ehrenbergi cytotypes by the almost entire absence of heterochromatin in acrocentric autosomes and the presence of heterochromatin arms iin subtelocentric autosomes. Nucleolar organizing regions were found mainly on three pairs of chromosomes, but some differences in their localization were revealed. Comparison of G-banded chromosomes showed, that most chromosomes have a similar pattern. The types of chromosomal rearrangemetns were revealed due to the banding methods.     

Ultraviolet radiation-absorbing mycosporine-like amino acids (MAAs) are acquired from their diet by medaka fish (Oryzias latipes) but not by SKH-1 hairless mice

To assess whether vertebrates can acquire, from their diet, ultraviolet radiation-absorbing mycosporine-like amino acids (MAAs), medaka fish and hairless mice were maintained for 150 and 130 days, respectively, on diets either including Mastocarpus stellatus (rich in MAAs) or the same diets without this red alga. In medaka, the MAAs palythine and asterina-330, present in trace quantities in the diet with added M. stellatus, were present in significantly greater quantities in the eyes of fish fed this diet than in the eyes of control fish. Only traces of MAAs were present in the skin of medaka fed the diet containing MAAs. Shinorine, the principal MAA in M. stellatus, was not found in any tissues of medaka, which raises questions about the specificity of transport of MAAs. In hairless mice, no dietary MAAs were found in the tissues of the eyes, skin, or liver after maintenance on the experimental diet. Low concentrations of shinorine were present only in the tissues of the small and large intestines. These results indicate that MAAs are acquired from their diet and translocated to superficial tissues by teleost fish, but that mammals may be incapable of such. Thus, dietary supplementation with MAAs may be useful in aquacultured species of fish, but MAAs as ‘dietary sunscreens' may not be an option for mammals, including humans. Nevertheless, our demonstration of the uptake of shinorine by human skin cancer cells in culture raises evolutionary questions regarding the organ specificity of the capacity for the cellular transport of MAAs.     

Direct Species Identification of Common Pathogenic Dermatophyte Fungi in Clinical Specimens by Semi-nested PCR and Restriction Fragment Length Polymorphism

OBJECTIVE: To seek a rapid and reliable molecular biology method to identify the common pathogenic dermatophyte fungi from clinical samples. METHOD: The genome DNA was extracted from cultured strains of seven common dermatophyte fungi species and part of each positive clinical specimen by microscopy. Intergenic spacer regions of ribosomal DNA (ITS) were amplified by semi-nested PCR (snPCR) with three universal primers (NS5, ITS1, and ITS4) for fungi. The amplified products were digested with two restriction endonucleases (BciT130 I, Dde I), the Restriction Fragment Length Polymorphism(RFLP). The rest of each clinical specimen was cultured in Sabouraud's Agar medium. Then the results of RFLP were compared with the traditional culture results. RESULTS: The digestion of seven common dermatophyte fungi produced seven different restriction profiles. Restriction profiles of 17 clinical specimens matched, respectively, to that of the cultured strains, and 14 profiles of the 17 ones matched the culture result completely. The coincidence was 100.0%. CONCLUSIONS: snPCR-RFLP analysis of intergenic spacer regions of ribosomal DNA is a valuable method of exactness and clarity for species identification of common dermatophyte fungi from clinical specimens.     

Revision of the bathyal cusk-eels of the genus Bassogigas (Ophidiidae) with description of a new species from off Guam, west Pacific Ocean

The bathyal genus Bassogigas (Teleostei: Ophidiidae) is revised based on 25 specimens, 18 from the west Atlantic Ocean and seven from the Indo-west Pacific Ocean. One specimen, from off Guam, west Pacific Ocean, represents a new species, Walker's cusk eel Bassogigas walkeri. The other 24 specimens all belong to the type species, Gills cusk eel Bassogigas gillii. A comparison between the Atlantic and the Indo-west Pacific Ocean specimens of B. gillii showed no differences in meristic and morphometric characters, but in two of the Indo-west Pacific Ocean specimens the sagittal otolith varied somewhat from the remaining specimens. The two Bassogigas species differ in the length of the lateral line, the number of scales in the midline of the body, the form of the median basibranchial tooth patches and in the thickness of the otolith.     

Does a type specimen necessarily or contingently belong to its species?

In a recent article, Alex Levine raises a paradox. It appears that, given some relatively uncontroversial premises about how a species term comes to refer to its species, a type specimen belongs necessarily and contingently to its species. According to Levine, this problem arises if species are individuals rather than natural kinds. I argue that the problem can be generalized: the problem also arises if species are kinds and type specimens are paradigmatic members used to baptize names for species. Indeed, the same problem arises with respect to kinds like gold and the samples used to ground names for them. After arguing that the paradox arises whether or not species are individuals, I attempt to show how the paradox can be resolved. Levine's argument that a type specimen belongs necessarily to its species is specious. The appeal of the argument stems from a failure to distinguish between two different modal statements concerning type specimens, one de dicto and the other de re. Type specimens belong contingently to their respective species. Even so, they can be known a priori to belong to them: hence, that a particular type specimen belongs to its species is an example of contingent a priori knowledge.     

我国华鳊属鱼类形态差异及其物种有效性的研究

为探讨华鳊属鱼类种间和种内居群间的形态差异及其物种有效性,采用多变量形态度量,结合传统分类方法,获得该属鱼类不同地理居群共224尾标本的32个测量性状和7个可数性状数据,并做主成分分析。在可数性状上,海南华鳊各居群间无差异,但明显不同于其他种类;四川华鳊与大眼华鳊差别明显,与伍氏华鳊之间也存在较大程度的差异;大眼华鳊的乌江和珠江居群之间存在显著差异,而伍氏华鳊各地理居群差异不明显。在可量性状上,四川华鳊和大眼华鳊的乌江居群都明显区别于其他种或居群,而大眼华鳊的珠江居群与伍氏华鳊之间以及海南华鳊各居群之间没有明显差异。对上述结果分析认为：四川华鳊和海南华鳊为两个有效的物种;大眼华鳊的乌江标本是一个未被描述的物种,而珠江居群和伍氏华鳊各居群应属同一个种。     

Algal colonization and its ecophysiology on the fine sculptures of terracotta monuments of Bishnupur,West Bengal,India

The blackish brown crust on the fine carvings of seven terracotta monuments of Bishnupur were studied with regards to the presence of algal species. Forty five taxa of micro algae belonging to 36 species of cyanobacteria, 5 species of chlorophyta and 4 species of bacillariophyta were found in crusts of these monuments. Among the algal groups, cyanobacteria were the dominant population growing in all the monuments with maximum diversity of 30 species in Rasmancha. Lyngbya corticicola, Scytonema schmidtii and Nostoc microscopicum were the major taxa in all crust samples. The absorption ratio of chlorophyll: carotenoid: MAAs (mycosporine-like amino acid substances) and scytonemin showed scytonemin and MAAs were more abundant than the chlorophyll pigment in all the crust. The MAAs and scytonemin of the crusts were identified as Mycosporine glycine having absorption maxima at 310 nm with retention time (RT) 4.1 min and scytonine having absorption maxima 386 nm with RT 3.2 min respectively. Among the major crust cyanobacteria cellular carbohydrate of N. microscopicum was maximum and lowest in S. schmidtii. However, the extra polymeric substance released to the medium was more in L. corticicola and so intense that the medium become coloured. The slimes of the major crust cyanobacteria plug the fine carvings of the terracotta structures which led to disfiguration of fine architecture, act as a biofilm that facilitated growth of fungi, lichens and subsequently to mosses and small plants leading to create creaks in the monuments and deteriorate these archeologically important monuments.     

Mycosporine-like amino acids in planktonic organisms living under different UV exposure conditions in Patagonian lakes

Mycosporine-like amino acids (MAAs) were studied in zooplanktonfrom 13 Argentinian lakes covering a broad range in altitude,maximum depth and physico-chemical properties of the water.Four to nine different MAAs (predominantly porphyra-334 andshinorine) were found in the copepods Boeckella gibbosa, B.gracilipes, B. meteoris and Parabroteas sarsi, and in the ciliateStentor amethystinus, while MAAs were undetectable in the cladoceranDaphnia middendorffiana. Among the different copepods, maximumMAA concentrations accounted for 0.25–1.31% of the dryweight, and contents were generally about three to seven times(up to 43 times) higher in the animals living in the clearestlakes compared to those occurring in low-UV systems. This variabilityin the content of MAAs was related to the lake altitude (r²= 0.71), and the fraction of the water column to which 1% ofthe surface UV radiation at 320 nm penetrated (r² = 0.57). Ourdata therefore underscore the role of MAAs as sunscreens todecrease the potential negative effects of solar radiation,but they also indicate that other environmental factors besidesUV transparency play a role in determining MAA concentrations.One lake was selected to obtain additional information on thequalitative composition of MAAs in seston of <100 µmbetween two sampling sites and over a 2 month study period (australsummer). Six different MAAs were detected in the samples, withporphyra-334 and palythine being predominant. In the copepodscollected simultaneously, there was low variation in MAA concentrationsbetween the two sites and over time. Thus, our results suggestthat under similar UV exposure conditions MAA contents of planktonicorganisms show low temporal variation.     

Finite element models predict cancellous apparent modulus when tissue modulus is scaled from specimen CT-attenuation

High-resolution architecture-based finite element models are commonly used for characterizing the mechanical behavior of cancellous bone. The vast majority of studies use homogeneous material properties to model trabecular tissue. The objectives of this study were to demonstrate that inhomogeneous finite element models that account for microcomputed tomography-measured tissue modulus variability more accurately predict the apparent stiffness of cancellous bone than homogeneous models, and to examine the sensitivity of an inhomogeneous model to the degree of tissue property variability. We tested five different material cases in finite element models of ten cancellous cubes in simulated uniaxial compression. Three of these cases were inhomogeneous and two were homogeneous. Four of these cases were unique to each specimen, and the remaining case had the same tissue modulus for all specimens. Results from all simulations were compared with measured elastic moduli from previous experiments. Tissue modulus variability for the most accurate of the three inhomogeneous models was then artificially increased to simulate the effects of non-linear CT-attenuation-modulus relationships. Uniqueness of individual models was more critical for model accuracy than level of inhomogeneity. Both homogeneous and inhomogeneous models that were unique to each specimen had at least 8% greater explanatory power for apparent modulus than models that applied the same material properties to all specimens. The explanatory power for apparent modulus of models with a tissue modulus coefficient of variation (COV) range of 21-31% was 13% greater than homogeneous models (COV=0). The results of this study indicate that inhomogenous finite element models that have tissue moduli unique to each specimen more accurately predict the elastic behavior of cancellous cubic specimens than models that have common tissue moduli between all specimens.     

New skull of Lesothosaurus (Dinosauria: Ornithischia) from the Upper Elliot Formation (Lower Jurassic) of southern Africa

A new ornithischian skull from the Elliot Formation of southern Africa is described. The specimen is compared in detail with the fabrosaurid Lesothosaurus diagnosticus. It actually shares many characters with specimens of the syntypes of this species or specimens referred to it. It is nevertheless not identical to any of these specimens and it is, moreover, remarkably larger than them. The possibility of attributing this specimen to a so far undescribed ‘large fabrosaur’ from the same formation is discussed. It is concluded that the specimen in question in this paper, while being ascribable to the genus Lesothosaurus, cannot be determined to a specific level until the existence of two fabrosaurid species in the ‘Stormberg Group’ is demonstrated and their range of morphological and size variation is properly appraised.