Hydroxyapatite biomaterial production from chicken (femur and beak) and fishbone waste through a chemical less method for Cd2+ removal from shipbuilding wastewater.

Hydroxyapatite (HAp) powder was produced from chicken (femur and beak) and fishbone wastes and used as a green adsorbent to decrease Cd2+ from aqueous media. The HAp powder was generated at 900 °C and characterized using physicochemical techniques. Chicken femur' HAp (16.72 m2/g) had a higher surface compared to chicken beak and fishbone ones. The solution pH was the most important parameter in removing Cd2+. The highest Cd2+ removal was achieved at pH 6, temperature of 25 °C, contact time of 80 min, and adsorbent mass of 2 g/L. The Cd2+ adsorption data fitted well with the quasi-second-order model in kinetics and the Freundlich model in isotherm. The highest adsorption capacity of Cd2+ using HAp-chicken femur, HAp-fish bone, and HAp-chicken beak was determined 22.94 mg/g, 21.54 mg/g, and 21.45 mg/g, respectively. The Cd2+ adsorption using HAp powder was a spontaneous and exothermic process and accidental collisions at the liquid-solid interface were reduced. The decrease of Cd2+ adsorption efficiency was not significant after multiple recovery steps of the desired powders. In addition to Cd2+, other parameters of real wastewater (shipbuilding industry) were reduced by the proposed adsorbents. The utilization of hydroxyapatite powder is expected to be a cheap and eco-friendly method for eliminating metals such as Cd2+.

Cephalopod beak sections used to trace mercury levels throughout the life of cephalopods: The giant warty squid Moroteuthopsis longimana as a case study.

Cephalopods represent an important pathway for mercury transfer through food webs. Due to the general difficulties in capturing oceanic squid, beaks found in the diet of top predators can be used to study their life-cycles and ecological role. Using upper beaks of the giant warty squid Moroteuthopsis longimana (major prey in the Southern Ocean), we describe a method to assess mercury concentrations along the life of cephalopods through the segmentary analysis of beak sections (i.e. tip of the rostrum and subsections along the hood). Distinct total mercury concentrations in the different subsections support that beaks can be used to study mercury levels in different periods of cephalopods' life-cycle. Mercury values in the anterior (1.3-7.9 µg kg-1 dw) and posterior (7.8-12.5 µg kg-1 dw) subsections of the hood reflect juvenile and adult stages, respectively. Furthermore, these results confirm that mercury bioaccumulates continuously throughout the individuals' life, with adults doubling their mercury concentrations to juveniles.

Long-term changes in habitat and trophic level of Southern Ocean squid in relation to environmental conditions.

Long-term studies of pelagic nekton in the Southern Ocean and their responses to ongoing environmental change are rare. Using stable isotope ratios measured in squid beaks recovered from diet samples of wandering albatrosses Diomedea exulans, we assessed decadal variation (from 1976 to 2016) in the habitat (δ13C) and trophic level (δ15N) of five important Southern Ocean squid species in relation to indices of environmental conditions-Southern Oscillation Index (SOI) and Southern Annular Mode (SAM). Based on δ13C values, corrected for the Suess effect, habitat had changed over the last 50 years for Taonius sp. B (Voss), Gonatus antarcticus, Galiteuthis glacialis and Histioteuthis atlantica but not Moroteuthopsis longimana. By comparison, mean δ15N values were similar across decades for all five species, suggesting minimal changes in trophic levels. Both SAM and SOI have increased in strength and frequency over the study period but, of the five species, only in Taonius sp. B (Voss) did these indices correlate with, δ13C and δ15N values, indicating direct relationships between environmental conditions, habitat and trophic level. The five cephalopod species therefore changed their habitats with changing environmental conditions over the last 50 years but maintained similar trophic levels. Hence, cephalopods are likely to remain important prey for top predators in Southern Ocean food webs, despite ongoing climate change.

Antarctic octopod beaks as proxy for mercury concentrations in soft tissues.

As the role of mercury is poorly known in Southern Ocean biota, the total mercury (T-Hg) concentrations were evaluated in upper/lower beaks, digestive gland, gills and mantle muscle of Adelieledone polymorpha and Pareledone turqueti, two of the most abundant octopod species around South Georgia. Beaks had the lowest T-Hg concentrations (A. polymorpha: [T-Hg]Upper = 27.2 ± 12.9 ng∙g-1 and [T-Hg]Lower = 27.5 ± 20.0 ng∙g-1; P. turqueti: [T-Hg]Upper = 34.6 ± 13.9 ng∙g-1 and [T-Hg]Lower = 56.8 ± 42.0 ng∙g-1), followed by gills and muscle. The highest values were recorded in the digestive gland (A. polymorpha: 251.6 ± 69.7 ng∙g-1; P. turqueti: 347.0 ± 177.0 ng∙g-1). Significant relationships were found between the concentrations of T-Hg in the beaks and muscle of A. polymorpha (T-Hg in muscle is 10 times higher than in beaks). This study shows that beaks can be used as proxy for T-Hg in muscle for some octopod species, and a helpful tool for estimating total Hg body burden from beaks.

Evaluation of Goose-beak Bone Particles for Dentoalveolar Reconstruction in Dogs.

BACKGROUND/AIM: Tooth extraction is a common procedure in dental clinics. Tooth extraction can destroy gingiva, alveolar bone, periodontal ligaments and cement. If dental sockets are left as extracted, it will result in loss of teeth, as well as voice and aesthetic problems. A natural hydroxyapatite (HA) bioceramic bone graft substitute developed from goose-beak bone particles (GBPs) was used for dentoalveolar reconstruction in a canine model. MATERIALS AND METHODS: Four adult (18-22 months old) male beagle dogs weighing 8.2-9.6 kg were included in the study. Eight alveolar extraction sockets in the four dogs were divided randomly into two groups and a split-mouth design was established; control group, socket filled with commercial synthetic HA; tested group, socket filled with granulated GBP. RESULTS: Micro-CT analysis and hematoxylin and eosin and Masson's trichrome staining of non-decalcified sections were undertaken. Examination revealed that dentoalveolar reconstruction was initiated from the periphery of the host bone, and newly formed bone was well integrated with the GBP. Bone apposition was observed at the edge of the host bone-GBP interface. CONCLUSION: A natural ceramic powder obtained from GBP is suitable for use in dentoalveolar reconstruction in dogs.

Hydrogen bond guidance and aromatic stacking drive liquid-liquid phase separation of intrinsically disordered histidine-rich peptides.

Liquid-liquid phase separation (LLPS) of intrinsically disordered proteins (IDPs) is involved in both intracellular membraneless organelles and extracellular tissues. Despite growing understanding of LLPS, molecular-level mechanisms behind this process are still not fully established. Here, we use histidine-rich squid beak proteins (HBPs) as model IDPs to shed light on molecular interactions governing LLPS. We show that LLPS of HBPs is mediated though specific modular repeats. The morphology of separated phases (liquid-like versus hydrogels) correlates with the repeats' hydrophobicity. Solution-state NMR indicates that LLPS is a multistep process initiated by deprotonation of histidine residues, followed by transient hydrogen bonding with tyrosine, and eventually by hydrophobic interactions. The microdroplets are stabilized by aromatic clustering of tyrosine residues exhibiting restricted molecular mobility in the nano-to-microsecond timescale according to solid-state NMR experiments. Our findings provide guidelines to rationally design pH-responsive peptides with LLPS ability for various applications, including bioinspired protocells and smart drug-delivery systems.

Show your beaks and we tell you what you eat: Different ecology in sympatric Antarctic benthic octopods under a climate change context.

Sympatry can lead to higher competition under climate change and other environmental pressures, including in South Georgia, Antarctica, where the two most common octopod species, Adelieledone polymorpha and Pareledone turqueti, occur side by side. Since cephalopods are typically elusive animals, the ecology of both species is poorly known. As beaks of cephalopods are recurrently found in top predator's stomachs, we studied the feeding ecology of both octopods through the evaluation of niche overlapping and specific beak adaptations that both species present. A multidisciplinary approach combining carbon (δ13C) and nitrogen (δ15N) stable isotope signatures, mercury (Hg) analysis and biomaterials' engineering techniques was applied to investigate the beaks. An isotopic niche overlap of 95.6% was recorded for the juvenile stages of both octopod species, dropping to 19.2% for the adult stages. Both A. polymorpha and P. turqueti inhabit benthic ecosystems around South Georgia throughout their lifecycles (δ13C: -19.21 ±â€¯1.87‰, mean ±â€¯SD for both species) but explore trophic niches partially different during adult life stages (δ15N: 7.01 ±â€¯0.40‰, in A. polymorpha, and 7.84 ±â€¯0.65‰, in P. turqueti). The beaks of A. polymorpha are less dense and significantly less stiff than in P. turqueti. Beaks showed lower mercury concentration relative to muscle (A. polymorpha - beaks: 0.052 ±â€¯0.009  µg g-1, muscle: 0.322 ±â€¯0.088  µg g-1; P. turqueti - beaks: 0.038 ±â€¯0.009  µg g-1; muscle: 0.434 ±â€¯0.128  µg g-1). Overall, both octopods exhibit similar habitats but different trophic niches, related to morphology/function of beaks. The high Hg concentrations in both octopods can have negative consequences on their top predators and may increase under the present climate change context.

Glucose-Responsive Peptide Coacervates with High Encapsulation Efficiency for Controlled Release of Insulin.

A new glucose-responsive insulin delivery system is fabricated using biomimetic peptide coacervates derived from the Humboldt squid (Dosidicus Gigas) beak. Both insulin and glucose oxidase are coencapsulated within coacervate microdroplets. The glucose oxidase quickly responds to increasing glucose levels to generate a local acidic environment, thereby rapidly triggering the dissociation of pH-sensitive coacervates to release the insulin cargo. The rate of insulin release is dependent on the glucose level, increases under hyperglycemic conditions, and decreases under normoglycemic conditions. This glucose responsiveness mimics pancreatic ß-cell function by releasing insulin according to glucose levels.

Spatial and trophic preferences of jumbo squid Dosidicus gigas (D'Orbigny, 1835) in the central Gulf of California: ecological inferences using stable isotopes.

RATIONALE: The jumbo squid Dosidicus gigas is a fishery resource of considerable economic and ecological importance in the Mexican Pacific. Studies on its habitat preferences are needed to understand recent fluctuations in the abundance and availability of the species. Stable isotope analysis allows us to infer ecological aspects such as spatial distribution and trophic preferences. METHODS: We used an isotope ratio mass spectrometer, automated for carbonate analysis, and coupled to an elemental analyzer, to determine the isotopic composition of statoliths (δ18 O and δ13 C values) and beaks (δ13 C and δ15 N values) from 219 individuals caught over two fishing seasons (2007 and 2009) off the coast of Santa Rosalía, in the central Gulf of California. We used these isotopic ratios to assess variation in spatial and trophic preferences by sex, size, and fishing season. RESULTS: In the 2009 group, we observed significant differences in statolith δ13 C values and beak δ13 C and δ15 N values between males and females. Between size groups, we observed significant differences in statolith δ18 O and δ13 C values in 2007 and in beak δ13 C and δ15 N values during both seasons. Both seasons were characterized by high overlap in δ18 O and δ13 C values between sexes and in 2009 between size groups. We observed low trophic overlap between sexes in 2009 and between size groups during both seasons. CONCLUSIONS: The isotopic ratios from statoliths and beaks indicate that D. gigas has changed its spatial and trophic preferences, a shift that is probably related to changes in the species' diet. This intraspecific variation in preferences could be related to characteristics such as size, which may influence squid distribution preferences.

Self-coacervation of modular squid beak proteins - a comparative study.

The beak of the Humboldt squid is a biocomposite material made solely of organic components - chitin and proteins - which exhibits 200-fold stiffness and hardness gradients from the soft base to the exceptionally hard tip (rostrum). The outstanding mechanical properties of the squid beak are achieved via controlled hydration and impregnation of the chitin-based scaffold by protein coacervates. Molecular-based understanding of these proteins is essential to mimic the natural beak material. Here, we present detailed studies of two histidine-rich beak proteins (HBP-1 and -2) that play central roles during beak bio-fabrication. We show that both proteins have the ability to self-coacervate, which is governed intrinsically by the sequence modularity of their C-terminus and extrinsically by pH and ionic strength. We demonstrate that HBPs possess dynamic structures in solution and achieve maximum folding in the coacervate state, and propose that their self-coacervation is driven by hydrophobic interactions following charge neutralization through salt-screening. Finally, we show that subtle differences in the modular repeats of HBPs result in significant changes in the rheological response of the coacervates. This knowledge may be exploited to design self-coacervating polypeptides for a wide range of engineering and biomedical applications, for example bio-inspired composite materials, smart hydrogels and adhesives, and biomedical implants.

The significance of cephalopod beaks in marine ecology studies: Can we use beaks for DNA analyses and mercury contamination assessment?

Cephalopod beaks found in the diet of predators have been a major source of scientific information. In this study, we evaluated the usefulness of DNA and contaminants analysis (total mercury - T-Hg) in cephalopod beaks in order to assess their applicability as tools in marine ecology studies. We concluded that, when applying DNA techniques to cephalopod beaks from Antarctic squid species, when using flesh attached to those beaks, it was possible to obtain DNA and to successfully identify cephalopod species; DNA was not found on the beaks themselves. This study also showed that it is possible to obtain information on T-Hg concentrations in beaks: the T-Hg concentrations found in the beaks were 6 to 46 times lower than in the flesh of the same cephalopod species. More research on the relationships of mercury concentrations in cephalopod beaks (and other tissues), intra- and inter-specifically, are needed in the future.

Infiltration of chitin by protein coacervates defines the squid beak mechanical gradient.

The beak of the jumbo squid Dosidicus gigas is a fascinating example of how seamlessly nature builds with mechanically mismatched materials. A 200-fold stiffness gradient begins in the hydrated chitin of the soft beak base and gradually increases to maximum stiffness in the dehydrated distal rostrum. Here, we combined RNA-Seq and proteomics to show that the beak contains two protein families. One family consists of chitin-binding proteins (DgCBPs) that physically join chitin chains, whereas the other family comprises highly modular histidine-rich proteins (DgHBPs). We propose that DgHBPs play multiple key roles during beak bioprocessing, first by forming concentrated coacervate solutions that diffuse into the DgCBP-chitin scaffold, and second by inducing crosslinking via an abundant GHG sequence motif. These processes generate spatially controlled desolvation, resulting in the impressive biomechanical gradient. Our findings provide novel molecular-scale strategies for designing functional gradient materials.

Hierarchical multiscale structure-property relationships of the red-bellied woodpecker (Melanerpes carolinus) beak.

We experimentally studied beaks of the red-bellied woodpecker to elucidate the hierarchical multiscale structure-property relationships. At the macroscale, the beak comprises three structural layers: an outer rhamphotheca layer (keratin sheath), a middle foam layer and an inner bony layer. The area fraction of each layer changes along the length of the beak giving rise to a varying constitutive behaviour similar to functionally graded materials. At the microscale, the rhamphotheca comprises keratin scales that are placed in an overlapping pattern; the middle foam layer has a porous structure; and the bony layer has a big centre cavity. At the nanoscale, a wavy gap between the keratin scales similar to a suture line was evidenced in the rhamphotheca; the middle foam layer joins two dissimilar materials; and mineralized collagen fibres were revealed in the inner bony layer. The nano- and micro-indentation tests revealed that the hardness (associated with the strength, modulus and stiffness) of the rhamphotheca layer (approx. 470 MPa for nano and approx. 320 MPa for micro) was two to three times less than that of the bony layer (approx. 1200 MPa for nano and approx. 630 MPa for micro). When compared to other birds (chicken, finch and toucan), the woodpecker's beak has more elongated keratin scales that can slide over each other thus admitting dissipation via shearing; has much less porosity in the bony layer thus strengthening the beak and focusing the stress wave; and has a wavy suture that admits local shearing at the nanoscale. The analysis of the woodpeckers' beaks provides some understanding of biological structural materials' mechanisms for energy absorption.

The potential role of oral pH in the persistence of Trichomonas gallinae in Cooper's Hawks (Accipiter cooperii).

Trichomoniasis, caused by the protozoan Trichomonas gallinae, affects a variety of species worldwide including avivorious raptors. Existing information suggests that the disease is most prevalent in young birds, and differential susceptibility to trichomoniasis among individuals in different age groups was documented in Cooper's Hawks (Accipiter cooperii) nesting in Tucson, Arizona. In that population, 85% of nestling Cooper's Hawks had T. gallinae in their oral cavity, compared to only 1% of breeding-age hawks. Trichomonads generally are sensitive to environmental pH and we explored the possibility that differences in oral pH may contribute to the differential prevalence of infection between age groups. We measured the pH of the fluid in the oral cavity in 375 Cooper's Hawks from three age groups (nestlings, fledglings, and breeding age) in Tucson, Arizona, in 2010 and 2011 and clinically tested for T. gallinae in a subsample of hawks. Oral pH of nestlings (∼ 6.8) was 7.3 times less acidic than in fledgling or breeding Cooper's Hawks (∼ 6.1). The incidence of T. gallinae was higher in nestlings (16%) than in either fledglings or breeding hawks (0%). Our findings indicate that oral pH becomes more acidic in Cooper's Hawks soon after they leave the nest. Trichomonas gallinae thrives when pH is between 6.5 and 7.5 (optimum 7.2), but is less viable in more acidic conditions. Higher levels of acidity in the oral cavity of fledglings and breeding Cooper's Hawks may reduce their susceptibility to trichomoniasis, and play a role in the differential prevalence of infection among age groups.

Bioinspired water-enhanced mechanical gradient nanocomposite films that mimic the architecture and properties of the squid beak.

Inspired by the water-enhanced mechanical gradient character of the squid beak, we herein report a nanocomposite that mimics both the architecture and properties of this interesting natural material. Similar to the squid beak, we have developed nanocomposites where the degree of cross-linking is controlled along the length of the film. In this study, we utilized tunicate cellulose nanocrystals as the nanofiller that are functionalized with allyl moieties. Using photoinduced thiol-ene chemistry, we have been able to cross-link the CNC nanofiller. In the dry state where strong CNC interactions can occur, only a small mechanical contrast is observed between the cross-linked and uncross-linked samples. However, when the films are exposed to water, which "switches off" the noncovalent CNC interactions, a significant mechanical contrast is observed between the same films. For example, at 20 wt % CNC (in the dry film), an increase in wet modulus from 60 to 300 MPa (∼500% increase) is observed after photoirradiation. Furthermore, we show that the wet modulus can be controlled by altering the UV exposure time which allows access to mechanical gradient films.

Chicken corneocyte cross-linked proteome.

Shotgun proteomic analysis was performed of epidermal scale, feather, beak and claw from the domestic chicken. To this end, the samples were separated first into solubilized and particulate fractions, the latter enriched in isopeptide cross-linking, by exhaustive extraction in sodium dodecyl sulfate under reducing conditions. Among the 205 proteins identified were 17 keratins (types α and ß), 51 involved in protein synthesis, 8 junctional, 8 histone, 5 heat shock, and 5 14-3-3 proteins. Considerable overlap among the beak, claw, feather, and scale samples was observed in protein profiles, but those from beak and claw were the most similar. Scale and feather profiles were the most distinctive, each exhibiting specific proteins. Less than 20% of the proteins were found only in the detergent-solubilized fraction, while 34-57% were found only in the particulate fraction, depending on the source, and the rest in both fractions. The results provide the first comprehensive analysis of the content of these cornified structures, reveal the efficient use of available proteins in conferring mechanical and chemical stability to them, and emphasize the importance of isopeptide cross-linking in avian epithelial cornification.

Candidate genes for carotenoid coloration in vertebrates and their expression profiles in the carotenoid-containing plumage and bill of a wild bird.

Carotenoid-based coloration has attracted much attention in evolutionary biology owing to its role in honest, condition-dependent signalling. Knowledge of the genetic pathways that regulate carotenoid coloration is crucial for an understanding of any trade-offs involved. We identified genes with potential roles in carotenoid coloration in vertebrates via (i) carotenoid uptake (SR-BI, CD36), (ii) binding and deposition (StAR1, MLN64, StAR4, StAR5, APOD, PLIN, GSTA2), and (iii) breakdown (BCO2, BCMO1). We examined the expression of these candidate loci in carotenoid-coloured tissues and several control tissues of the red-billed quelea (Quelea quelea), a species that exhibits a male breeding plumage colour polymorphism and sexually dimorphic variation in bill colour. All of the candidate genes except StAR1 were expressed in both the plumage and bill of queleas, indicating a potential role in carotenoid coloration in the quelea. However, no differences in the relative expression of any of the genes were found among the quelea carotenoid phenotypes, suggesting that other genes control the polymorphic and sexually dimorphic variation in carotenoid coloration observed in this species. Our identification of a number of potential carotenoid genes in different functional categories provides a critical starting point for future work on carotenoid colour regulation in vertebrate taxa.

Determination and validation of the elastic moduli of small and complex biological samples: bone and keratin in bird beaks.

In recent years, there has been a surge in the development of finite-element (FE) models aimed at testing biological hypotheses. For example, recent modelling efforts suggested that the beak in Darwin's finches probably evolved in response to fracture avoidance. However, knowledge of the material properties of the structures involved is crucial for any model. For many biological structures, these data are not available and may be difficult to obtain experimentally given the complex nature of biological structures. Beaks are interesting as they appear to be highly optimized in some cases. In order to understand the biomechanics of this small and complex structure, we have been developing FE models that take into account the bilayered structure of the beak consisting of bone and keratin. Here, we present the results of efforts related to the determination and validation of the elastic modulus of bone and keratin in bird beaks. The elastic moduli of fresh and dried samples were obtained using a novel double-indentation technique and through an inverse analysis. A bending experiment is used for the inverse analysis and the validation of the measurements. The out-of-plane displacements during loading are measured using digital speckle pattern interferometry.

Comparative study of the mechanical properties, micro-structure, and composition of the cranial and beak bones of the great spotted woodpecker and the lark bird.

Woodpeckers are well able to resist head injury during repeated high speed impacts at 6-7 m s⁻¹ with decelerations up to 1000 g. This study was designed to compare the mechanical properties, microstructures and compositions of cranial bone and beak bone of great spotted woodpecker (Dendrocopos major) and the Mongolian sky lark (Melanocorypha mongolica). Microstructures were observed using micro-computed tomography and scanning electron microscopy and their compositions were characterized by X-ray powder diffraction and Fourier-transform infrared spectroscopy. Under high stress, the cranial bone and the beak of the woodpecker exhibited distinctive mechanical features, which were associated with differences in micro-structure and composition, compared with those of the lark. Evolutionary optimization of bone micro-structure has enabled functional adaptation to the woodpecker's specific lifestyle. Its characteristic micro-structure efficiently avoids head impact injury and may provide potential clues to the prevention of brain injury using bio-inspired designs of shock-absorbing materials.