Structural alterations in scales of Capoeta capoeta (Güldenstädt, 1773) from heavy metal-contaminated river water.

Heavy metal pollution, especially in freshwater, is a serious problem for aquatic organisms and human health. In this study, the scales of Capoeta capoeta living in the Karasu River (Turkey), which is estimated to be contaminated with pollutants, especially heavy metals, were examined for structural anomalies. Two stations on the river were selected for this purpose. Fish and surface water samples were taken at the stations. The heavy metal analyses were carried out in the water and the fish tissue. Heavy metal pollution was detected in the surface water. It was also observed that some heavy metals (As, Cu, Cd, Cr, Mn, Pb, Ni, Zn) accumulate in the fish tissue. Significant structural differences were observed on the dorsal surface of the scales, such as interrupted primary radii, damaged circuli, damaged focus, damaged anterior scale margin, broken focus, deformed scale structure, scattered chromatophores, dilatation of primary radii, loss of focus, damaged annuli, symmetry shift in the lateral line canal, eroded circuli, damaged posterior scale margin, double focus, branching in the primary radii, asymmetric circuli, incomplete annuli and interrupted secondary radii in each of the fish collected from the contaminated site. Heavy metals are suspected to be responsible for the structural anomalies in the scales. Based on these observations, it can be said that fish scales can be used as an effective indicator of water quality.

Effects of the extraction temperatures on the protein contents, gelatin purities, physicochemical properties, and functional properties of tilapia scale gelatins.

Herein, the effects of the extraction temperatures (45, 55, 65, 75, and 85 °C) on the protein contents, gelatin purities, physicochemical properties, and functional properties of tilapia scale gelatins were studied. Among these temperatures, 65 °C was the best extraction temperature to obtain good production yield (16.0 % ± 0.3 %), good protein contents (excellent total amino acid composition of 94.20 ± 0.76 g/100 g of gelatin, the highest hydrophobic amino acids percentage of 32.68 ± 0.24 g/100 g of gelatin), appropriate ATR-FTIR spectra peaks (Amides A, B, I, II, and III), high ß-sheet percentage (38.2 % ± 1.3 %), the highest purity of the gelatin structures (280, 140, and 125 kDa), the lowest nanoparticle sizes in atomic force microscopy results, the highest water-holding capacity (25.3 % ± 0.9 %), the highest fat-binding capacity (16.9 % ± 0.1 %), high foaming properties (foaming capacity of 151.7 % ± 7.6 % and foaming stability of 145.8 % ± 6.3 %), the lowest interfacial tension (2.1 ± 0.2 mN/m), the lowest emulsifying activity index (16.5 % ± 0.9 %), the highest emulsifying stability index (88.9 % ± 5.1 %), highest emulsion viscosity (1462 ± 17 mPa·s at the rotary speed of 6 rpm), the lowest initial droplet sizes, and lowest emulsion creaming index (6.9 % ± 0.7 %). This work provided a useful guide to choosing extraction temperature for gelatin extraction and a useful theory on the relationship between compositions and properties of a protein sample.

Biomimetic Mineralized Hydroxyapatite-Fish-Scale Collagen/Chitosan Nanofibrous Membranes Promote Osteogenesis for Periodontal Tissue Regeneration.

Commercial mammalian collagen-based membranes used for guided tissue regeneration (GTR) in periodontal defect repair still face significant challenges, including ethical concerns, cost-effectiveness, and limited capacity for periodontal bone regeneration. Herein, an enhanced biomimetic mineralized hydroxyapatite (HAp)-fish-scale collagen (FCOL)/chitosan (CS) nanofibrous membrane was developed. Specifically, eco-friendly and biocompatible collagen extracted from grass carp fish scales was co-electrospun with CS to produce a biomimetic extracellular matrix membrane. An enhanced biomimetic mineralized HAp coating provided abundant active calcium and phosphate sites, which promoted cell osteogenic differentiation, and showed greater in vivo absorption. In vitro experiments demonstrated that the HAp-FCOL/CS membranes exhibited desirable properties with no cytotoxicity, provided a mimetic microenvironment for stem cell recruitment, and induced periodontal ligament cell osteogenic differentiation. In rat periodontal defects, HAp-FCOL/CS membranes significantly promoted new periodontal bone formation and regeneration. The results of this study indicate that low-cost, eco-friendly, and biomimetic HAp-FCOL/CS membranes could be promising alternatives to GTR membranes for periodontal regeneration in the clinic.

Decoupled evolution of ventral and dorsal scales in agamid lizards: ventral keels are associated with arboreality.

Arboreality has evolved in all major vertebrate lineages and is often associated with morphological adaptations and increased diversification concomitant with accessing novel niche space. In squamate reptiles, foot, claw, and tail morphology are well-studied adaptations shown to be associated with transitions to arboreality. Here, we examined a less well understood trait-the keeled scale-in relation to microhabitat, climate, and diversification dynamics across a diverse lizard radiation, Agamidae. We found that the ancestral agamid had keeled dorsal but not ventral scales; further, dorsal and ventral keels are evolutionarily decoupled. Ventral keeled scales evolved repeatedly in association with arboreality and may be advantageous in reducing wear or by promoting interlocking when climbing. We did not find an association between keeled scales and diversification, suggesting keels do not allow finer-scale microhabitat partitioning observed in other arboreal-associated traits. We additionally found a relationship between keeled ventral scales and precipitation in terrestrial species where we posit that the keels may function to reduce scale degradation. Our results suggest that keeled ventral scales facilitated transitions to arboreality across agamid lizards, and highlight a need for future studies that explore their biomechanical function in relation to microhabitat and climate.

The effect of shortfin mako shark skin at the reattachment of a separated turbulent boundary layer.

This smooth flat experimental study investigates the capability of mako shark scales to control flow separation when placed downstream of the onset of turbulent boundary layer separation and within the reattachment region. The objective of the study is to validate the hypothesis that the shark scales' bristling and recoiling would prevent the flow separation on the flank region (the fastest flow region) of the shark. A rotating cylinder was used to induce an adverse pressure gradient over a flat plate to produce a region of separated flow where the shark skin specimen was mounted. Two types of mako shark scales (flank (B2) and between flank and dorsal fin (B1)) were positioned in the preferred flow direction on a flat plate. The B2 scales are slender, 200µm tall, and can bristle up to 50°. In contrast, B1 scales are wider, shorter, and can bristle at 30°. The bristling angle and shape are the main mechanisms by which the scales act to inhibit flow from moving upstream near the wall. Thus, the difference in the bristling angles and structures of the scales is attributed to the fact that the B2 scales function in a thicker boundary layer (behind the shark's gills) where they must bristle sufficiently high into the boundary layer to control the flow separation, and because the adverse pressure gradient in this region is higher where flow separation is more likely. The scales are placed in the reattachment region to elucidate their ability to control and reattach an already separated turbulent flow. The results show that B2 scales placed in the reattachment region reduce the size of the turbulent separation bubble and decrease the turbulent kinetic energy, while B1 scales have the opposite effect.

Scale-based landmark for internal organs assessments of common carp (Cyprinus carpio) during digital radiography and ultrasonography imaging.

Medical imaging techniques such as digital radiography and ultrasonography are non-invasive and provide precise results for examining internal organs and structures within fish. Their effectiveness can be further enhanced by using body parts like scales as markers for the organs beneath them. This study utilized the number of scales as landmarks in digital radiography and ultrasonography to non-invasively evaluate the muscles, bones, and images of internal and reproductive organs of common carp (Cyprinus carpio). Digital radiography was performed in the dorsoventral and lateral views of the fish, whereas ultrasonography was conducted in longitudinal and transverse views on sequence scale numbers with brightness and colour Doppler-modes. Digital radiography of the common carp revealed the whole-body morphology, including the bony parts from the head, pectoral fins, dorsal fins, pelvic fins, anal fins, and vertebrae to the tail that appeared radiopaque. Internal organs were also observed, with the swim bladder and heart appeared radiolucent, while the intestines, liver, testes, and ovaries appeared radiopaque. Ultrasonography in brightness mode displayed the digestive organs, reproductive organs, and muscle thickness. Additionally, colour Doppler mode demonstrated blood flow within the heart's ventricle.

Scale Characteristics of Six Fish Species of the Genus Cyprinion (Teleostei: Cypriniformes): A Microscopic Analysis.

This study investigated the morphological characteristics of scales in six Cyprinion species, using light and scanning electron microscopy focusing on key features such as scale type, key scales, lateral line scales, radius/radii, rostral margin, focus, circuli, lepidonts, tubercles, and scale indices. The research analyzed the scales using ultramicroscopy and light microscopy imaging, categorizing them based on size classes and body regions. The morphological variations in scale characteristics were examined across different species, regions, and size classes. Notable findings included the tetra-sectioned form of scales, representing a unique characteristic of the Cyprinion genus. Morphological changes in scale features were observed with fish growth, particularly in the overall shape, focus shape, and size. Quantitative analysis revealed variations in average relative scale length and width among different species, regions, and size classes. The study utilized canonical discriminant analysis for multivariate assessment, classifying the species into distinct groups based on morphometric indices. The findings contribute to the understanding of scale morphology in Cyprinion species and exploring morphological variation between the examined species.

3D printing of fish-scale derived hydroxyapatite/chitosan/PCL scaffold for bone tissue engineering.

In the field of bone tissue repair, the treatment of bone defects has always posed a significant challenge. In recent years, the advancement of bone tissue engineering and regenerative medicine has sparked great interest in the development of innovative bone grafting materials. In this study, a novel hydroxyapatite (HA) material was successfully prepared and comprehensively characterized. Antimicrobial experiments and biological evaluations were conducted to determine its efficacy. Based on the aforementioned research findings, 3D printing technology was employed to fabricate HA/chitosan (CS)/ polycaprolactone (PCL) scaffolds. The composition of the scaffold materials was confirmed through X-ray diffractometer (XRD) and Fourier Transform Infrared Spectroscopy (FT-IR) tests, while the influence of different HA ratios on the scaffold surface morphology was observed. Additionally, antimicrobial experiments demonstrated the favorable antimicrobial activity of the scaffolds containing 30%HA + 5%CS + PCL. Furthermore, the water contact angle measurements confirmed the superhydrophilicity of the scaffolds. Finally, the excellent bioactivity and ability to promote tissue regeneration of the scaffolds were further confirmed by in vitro and in vivo experiments. This study provides new options for future repair and regeneration of bone tissue and clinical applications.

Digestion and absorption characteristics of iron-chelating silver carp scale collagen peptide and insights into their chelation mechanism.

Iron deficiency is widespread throughout the world, supplementing sufficient iron or improving the bioavailability of iron is the fundamental strategy to solve the problem of iron scarcity. Herein, we explored a new form of iron supplement, iron chelates of silver carp scales (SCSCP-Fe) were prepared from collagen peptide of silver carp scales (SCSCP) and FeCl2·4H2O, the effects of external environment and simulated gastrointestinal digestive environment on the stability of SCSCP-Fe and the structural changes of peptide iron chelates during digestion were investigated. The results of in vitro iron absorption promotion showed that the iron bioavailability of SCSCP-Fe was higher than that of FeSO4. Two potential high iron chelating peptides DTSGGYDEY (DY) and LQGSNEIEIR (LR) were screened and synthesized from the SCSCP sequence by molecular dynamics and LC-MS/MS techniques. The FTIR results displayed that the binding sites of DY and LR for Fe2+ were the carboxyl group, the amino group, and the nitrogen atom on the amide group on the peptide. ITC results indicated that the chelation reactions of DY and LR with Fe2+ were mainly dominated by electrostatic interactions, forming chelates in stoichiometric ratios of 1:2 and 1:1, respectively. Both DY and LR had a certain ability to promote iron absorption. The transport of DY-Fe chelate may be a combination of the three pathways: PepT1 vector pathway, cell bypass, and endocytosis, while LR-Fe chelate was dominated by bivalent metal ion transporters. This study is expected to provide theoretical reference and technical support for the high-value utilization of silver carp scales and the development of novel iron supplements.

Uncertainty analysis of Altantic salmon fish scale's acoustic impedance using 30 MHz C-Scan measurements.

Understanding the biomechanics of fish scales is crucial for their survival and adaptation. Ultrasonic C-scan measurements offer a promising tool for non-invasive characterization, however, existing literature lacks uncertainty analysis while evaluating acoustic impedance. This article presents an innovative integration of uncertainty into the analytical framework for estimating stochastic specific acoustic impedance of salmon fish scale through ultrasonic C-scans. In this study, the various types of uncertainties arising due to variation in biological structures and aging, measurement errors, and analytical noises are combined together in the form of uncertain reflectance. This uncertain reflectance possesses a distribution which is derived using a theory of waves by assuming suitable stochasticity in wavenumber. This distribution helps in development of a stochastic-specific acoustic impedance map of the scales which demonstrates the possible deviations of impedance from mean value depending on uncertainties. Furthermore, maximal overlap discrete wavelet transform is employed for efficient time-frequency deconvolution and Kriging for spatial data interpolation to enhance the robustness of the impedance map, especially in scenarios with limited data. The framework is validated by accurately estimating the specific acoustic impedance of well-known materials like a pair of target medium (polyvinylidene fluoride) and reference medium (polyimide), achieving over 90% accuracy. Moreover, the accuracy of the framework is found superior when compared with an established approach in the literature. Applying the framework to salmon fish scales, we obtain an average specific acoustic impedance of 3.1 MRayl along with a stochastic map visualizing the potential variations arising from uncertainties. Overall, this work paves the way for more accurate and robust studies in fish scale biomechanics by incorporating a comprehensive uncertainty analysis framework.

Eco-friendly engineering of micro composite-based hydroxyapatite bio crystal and polyaniline for high removal of OG dye from wastewater: Adsorption mechanism and RSM@BBD optimization.

The presence of harmful substances such as dyes in water systems poses a direct threat to the quality of people's lives and other organisms living in the ecosystem. Orange G (OG) is considered a hazardous dye. The existing paper attempts to evaluate a low-cost adsorbent for the effective removal of OG dye. The developed adsorbent Polyaniline@Hydroxyapatite extracted from Cilus Gilberti fish Scale (PANI@FHAP) was elaborated through the application of the in situ chemical polymerization method to incorporate PANI on the surface of naturally extracted hydroxyapatite FHAP. The good synthesis of PANI@FHAP was evaluated through multiple techniques including X-ray diffraction (XRD), Scanning electron microscopy coupled with energy dispersive X-ray spectrometry (SEM/EDS), Fourier Transforms Infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA) coupled with thermal differential analysis (DTA) analysis. The results reveal a highly ordered disposition of PANI chains on FHAP, resulting in a well-coated FHAP in the PANI matrix. Furthermore, the presence of functional groups on the surface of PANI such as amine (-NH2) and imine (=NH) groups would facilitate the removal of OG dye from contaminated water. The adsorption of OG onto PANI@FHAP was conducted in batch mode and optimized through response surface methodology coupled with box-Behnken design (RSM/BBD) to investigate the effect of time, adsorbent dose, and initial concentration. The outcomes proved that OG adsorption follows a quadratic model (R2 = 0.989). The kinetic study revealed that the adsorption of OG fits the pseudo-second-order model. On the other hand, the isotherm study declared that the Freundlich model is best suited to the description of OG adsorption. For thermodynamic study, the adsorption of OG is spontaneous in nature and exothermic. Furthermore, the regeneration-reusability study indicates that PANI@FHAP could be regenerated and reused up to five successive cycles. Based on the FTIR spectrum of PANI@FHAP after OG adsorption, the mechanism governing OG adsorption is predominantly driven by π-π interaction, electrostatic interaction, and hydrogen bonding interactions. The obtained results suppose that PANI@FHAP adsorbent can be a competitive material in large-scale applications.

Hydrodynamic Function of the Slimy and Scaly Surfaces of Teleost Fishes.

The scales and skin mucus of bony fishes are both proposed to have a role in beneficially modifying the hydrodynamics of water flow over the body surface. However, it has been challenging to provide direct experimental evidence that tests how mucus and fish scales change the boundary layer in part due to the difficulties in working with live animal tissue and difficulty directly imaging the boundary layer. In this manuscript, we use direct imaging and flow tracking within the boundary layer to compare boundary layer dynamics over surfaces of fish skin with mucus, without mucus, and a flat control surface. Our direct measurements of boundary layer flows for these three different conditions are repeated for two different species, bluegill sunfish (Lepomis macrochirus) and blue tilapia (Oreochromis aureus). Our goals are to understand if mucus and scales reduce drag, shed light on mechanisms underlying drag reduction, compare these results between species, and evaluate the relative contributions to hydrodynamic function for both mucus and scales. We use our measurements of boundary layer flow to calculate shear stress (proportional to friction drag), and we find that mucus reduces drag overall by reducing the velocity gradient near the skin surface. Both bluegill and tilapia showed similar patterns of surface velocity reduction. We also note that scales alone do not appear to reduce drag, but that mucus may reduce friction drag up to 50% compared to scaled surfaces without mucus or flat controls.

The actin cytoskeleton plays multiple roles in structural colour formation in butterfly wing scales.

Vivid structural colours in butterflies are caused by photonic nanostructures scattering light. Structural colours evolved for numerous biological signalling functions and have important technological applications. Optically, such structures are well understood, however insight into their development in vivo remains scarce. We show that actin is intimately involved in structural colour formation in butterfly wing scales. Using comparisons between iridescent (structurally coloured) and non-iridescent scales in adult and developing H. sara, we show that iridescent scales have more densely packed actin bundles leading to an increased density of reflective ridges. Super-resolution microscopy across three distantly related butterfly species reveals that actin is repeatedly re-arranged during scale development and crucially when the optical nanostructures are forming. Furthermore, actin perturbation experiments at these later developmental stages resulted in near total loss of structural colour in H. sara. Overall, this shows that actin plays a vital and direct templating role during structural colour formation in butterfly scales, providing ridge patterning mechanisms that are likely universal across lepidoptera.

The age and growth information of a ctenoid scale fossil from the Upper Cretaceous Nenjiang Formation in Songliao Basin, China.

The study of morphological characteristics and growth information in fish scales is a crucial component of modern fishery biological research, while it has been less studied in fossil materials. This paper presents a detailed morphological description and growth analysis of a fossil ctenoid scale obtained from the Upper Cretaceous Campanian lacustrine deposits in northeastern China. The morphological features of this fossil scale are well-preserved and consistent with the structures found in ctenoid scales of extant fish species and display prominent ring ornamentation radiating outward from the central focus, with grooves intersecting the rings. A comparative analysis of the morphological characteristics between the fossil ctenoid scale and those well-studied extant fish Mugilidae allows us to explore the applicability of modern fishery biological research methods to the field of fossil scales. The scale length, scale width, the vertical distance from the focus to the apex of the scale, and the total number of radii have been measured. The age of the fish that possessed this ctenoid scale has been estimated by carefully counting the annuli, suggesting an age equal to or more than seven years. The distribution of growth rings on the scale potentially reflects the warm paleoclimatic condition and fish-friendly paleoenvironment prevalent during that period. This paper, moreover, serves as a notable application of fishery biological methods in the examination of fossil materials.

Cellular structure of dinosaur scales reveals retention of reptile-type skin during the evolutionary transition to feathers.

Fossil feathers have transformed our understanding of integumentary evolution in vertebrates. The evolution of feathers is associated with novel skin ultrastructures, but the fossil record of these changes is poor and thus the critical transition from scaled to feathered skin is poorly understood. Here we shed light on this issue using preserved skin in the non-avian feathered dinosaur Psittacosaurus. Skin in the non-feathered, scaled torso is three-dimensionally replicated in silica and preserves epidermal layers, corneocytes and melanosomes. The morphology of the preserved stratum corneum is consistent with an original composition rich in corneous beta proteins, rather than (alpha-) keratins as in the feathered skin of birds. The stratum corneum is relatively thin in the ventral torso compared to extant quadrupedal reptiles, reflecting a reduced demand for mechanical protection in an elevated bipedal stance. The distribution of the melanosomes in the fossil skin is consistent with melanin-based colouration in extant crocodilians. Collectively, the fossil evidence supports partitioning of skin development in Psittacosaurus: a reptile-type condition in non-feathered regions and an avian-like condition in feathered regions. Retention of reptile-type skin in non-feathered regions would have ensured essential skin functions during the early, experimental stages of feather evolution.

Spatio-temporal expression patterns of glycine-rich beta proteins and cysteine-rich beta proteins in setae development of Gekko japonicus.

BACKGROUND: Setae on the pad lamellae of the Japanese gecko Gekko japonicus (Schlegel, 1836), a vital epidermal derivative, are primarily composed of cornified beta-proteins (CBPs) and play a pivotal role in adhesion and climbing. The amino acid composition of CBPs might be a determining factor influencing their functional properties. However, the molecular mechanisms governed by CBP genes with diverse amino acid compositions in setae development remain unexplored. RESULTS: Based on RNA-seq analyses, this study confirmed that all G. japonicus CBPs (GjCBPs) are involved in setae formation. Cysteine-rich CBPs encoding genes (ge-cprp-17 to ge-cprp-26) and glycine-rich CBPs encoding genes (ge-gprp-17 to ge-gprp-22) were haphazardly selected, with quantitative real-time PCR revealing their expression patterns in embryonic pad lamellae and dorsal epidermis. It is inferred that glycine-rich CBPs are integral to the formation of both dorsal scales and lamellar setae, cysteine-rich CBPs are primarily associated with setae development. Additionally, fluorescence in situ hybridization revealed spatiotemporal differences in the expression of a glycine-rich CBP encoding gene (ge-gprp-19) and a cysteine-rich CBP encoding gene (ge-cprp-17) during dorsal scales and/or lamellar development. CONCLUSIONS: All 66 CBPs are involved in the formation of setae. Glycine-rich CBPs hold a significant role in the development of dorsal scales and lamellar setae, whereas most cysteine-rich CBPs appear to be essential components of G. japonicus setae. Even GjCBPs with similar amino acid compositions may play diverse functions. The clear spatio-temporal expression differences between the glycine-rich and cysteine-rich CBP encoding genes during epidermal scale and/or setae formation were observed. Embryonic developmental stages 39 to 42 emerged as crucial phases for setae development. These findings lay the groundwork for deeper investigation into the function of GjCBPs in the development of G. japonicus setae.

Effect and mechanism of natural composite hydrogel from fish scale intercellular matrix on diabetic chronic wound repair.

Diabetes mellitus is a chronic metabolic disease with prolonged low-grade inflammation and impaired cellular function, leading to poor wound healing. The treatment of diabetic wounds remains challenging due to the complex wound microenvironment. In view of the prominence of fish scales in traditional Chinese medicine and their wide application in modern medicine, we isolated the intercellular components in the scales of sea bass, obtained a natural composite hydrogel, fish scales gel (FSG), and applied it to diabetic chronic wounds. FSG was rich in collagen-like proteins, and possessed low-temperature gelation properties. In vitro, FSG was biocompatible and promoted fibroblast proliferation by approximately 40 %, endothelial cell migration by approximately 20 % and activated the M1 macrophages. In addition, FSG restored the function of fibroblasts and vascular endothelial cells damaged by high glucose. Importantly, FSG normalized the acute inflammatory response to impaired macrophages in a high-glucose microenvironment. Transcriptome analysis implies that this mechanism may involve enhanced cell signaling and cellular communication, improved sensitivity to cytokines, and activation of the TNF signaling pathway. Animal experiments confirmed that FSG significantly improved wound closure by approximately 15 % in diabetic rats, showing similar effects to acute wounds. In conclusion, the regulation of multiple cellular functions by FSG, especially the counterintuitive ability to induce acute inflammation, promoted diabetic wound healing and provides a novel therapeutic strategy for wound repair in diabetic patients.

Zebrafish Scale Regeneration In Toto and Ex Vivo Culture of Scales.

Skeletal diseases are often complex in their etiology and affect millions of people worldwide. Due to the aging population, there is a need for new therapeutics that could ease the burden on healthcare systems. As these diseases are complex, it is difficult and expensive to accurately model bone pathophysiology in a lab setting. The challenge for the field is to establish a cost-effective, biologically relevant platform for modeling bone disease that can be used to test potential therapeutic compounds. Such a platform should ideally allow dynamic visualization of cell behaviors of bone-building osteoblasts and bone-degrading osteoclasts acting in their mineralized matrix environment. Zebrafish are increasingly used as models due to the availability of genetic tools, including transgenic reporter lines, and the fact that some skeletal tissues (including the scales) remain translucent to adulthood, allowing dynamic imaging options. Since zebrafish scales have both osteoblasts and osteoclasts and are highly abundant, they provide an easily accessible and abundantly available resource of independent bone units. Moreover, once removed, adult zebrafish scales fully regenerate, therefore offering a way to study the spatiotemporal growth of mineralized tissue in vivo. Here, we detail protocols for harvesting and tracking the regeneration of the scales. Lastly, a protocol for stable culture of scales ex vivo for a week and following the healing response after controlled damage to the mineralized matrix of the scale over time is also presented.

Scale morphology variability in cyprinid fishes and its significance in taxonomy using light and scanning electron microscopy: A case study of the genus Garra Hamilton, 1822 (Teleostei: Cyprinidae).

To evaluate scale morphological variability (shape, size, topological macro- and microstructures, ornamentation patterns) of cyprinid fishes, nine species of the genus Garra were selected and their scales from five different body regions were studied by using light and scanning electron microscopy. The scales of the examined species were thin with a central or antero-centrally positioned focus, with no cteni in the posterior part. In addition to these typical characteristics, some morphological variation was observed in the overall shape (irregular round, true oval, round-triangular, irregular pentagonal, hexagonal, irregular hexagonal, pentagonal, ovoid), and the focus shape. These variations were mainly dependent on the fish lengths and the flank region. Morphological analysis clustered the examined species into two distinct groups. Group I consisted of G. amirhosseini and G. gymnothorax, while Group II has three subgroups, which include G. persica and G. mondica (subgroup I), G. meymehensis, G. rossica, G. nudiventris, G. hormuzensis (subgroup II), and G. rufa (subgroup III). The grouping of the studied Garra species based on the current scale morphological characters and the molecular data was only consistent for G. rossica and G. nudiventris. In addition, in the phylogenetic tree, G. persica, G. mondica, G. amirhosseini, and G. hormuzensis formed a distinct clade. However, these species did not represent close relationships in the dendrogram obtained from the scale morphology. A possible explanation why the grouping of the studied Garra species based on their scale morphological characters does not match their phylogenetic relationships is that most of the scale morphological traits vary depending on the fish size and the location of the scales on the flank. Therefore, except for some traits, that is, central or antero-centrally positioned focus, having no cteni, a specific sectioned form so called "tetra-sectioned" type, the other examined variables are not useful enough to be used in the taxonomic study of the examined cyprinid fishes. Therefore, scale morphological characters should be used carefully for taxonomic purposes. RESEARCH HIGHLIGHTS: Shape, size, topological macro- and microstructures, and ornamentation patterns of nine species of the genus Garra were studied by using light and scanning electron microscopy. The scales were thin with a central or antero-centrally positioned focus, with no cteni in the posterior part. Morphological variation was observed in the overall shape (irregular round, true oval, round-triangular, irregular pentagonal, hexagonal, irregular hexagonal, pentagonal, and ovoid), and the focus shape. The grouping of Garra species based on the current scale morphological characters and the molecular data was only consistent for G. rossica and G. nudiventris.

A diadectid skin impression and its implications for the evolutionary origin of epidermal scales.

Corneous skin appendages are not only common and diverse in crown-group amniotes but also present in some modern amphibians. This raises the still unresolved question of whether the ability to form corneous skin appendages is an apomorphy of a common ancestor of amphibians and amniotes or evolved independently in both groups. So far, there is no palaeontological contribution to the issue owing to the lack of keratin soft tissue preservation in Palaeozoic anamniotes. New data are provided by a recently discovered ichnofossil specimen from the early Permian of Poland that shows monospecific tetrapod footprints associated with a partial scaly body impression. The traces can be unambiguously attributed to diadectids and are interpreted as the globally first evidence of horned scales in tetrapods close to the origin of amniotes. Taking hitherto little-noticed scaly skin impressions of lepospondyl stem amniotes from the early Permian of Germany into account, the possibility has to be considered that the evolutionary origin of epidermal scales deeply roots among anamniotes.