Elimination of endocrine disrupting phenolic compounds via feathers and claws in seabirds moulting in the Baltic and Russian Arctic.

This paper investigates the effectiveness of phenol derivatives removal from bird organisms via claws and remiges, and performs a preliminary assessment of the usefulness of these epidermal products for environmental biomonitoring and estimating bird exposure levels. Concentrations of bisphenol A (BPA) and alkylphenols: 4-tert-octylphenol (4-t-OP) and 4-nonylphenol (4-NP) were determined in claws and remiges of long-tailed ducks Clangula hyemalis and razorbills Alca torda, obtained during a by-catch in the winter period (2014-2016) in the Southern Baltic region. For razorbills, the Baltic is a permanent habitat, while long-tailed ducks are migratory and stay in the Southern Baltic only during the non-breeding season. Their remiges are replaced in the Arctic seas of Siberia. The removal of phenol derivatives, depending on the compound and the epidermal product, ranges from 12 % to 34 %. Among these compounds, in both bird species, the highest degree of elimination was observed for 4-NP in remiges (<0.1-656.0 ng.g-1 dw) as well as claws (<0.1-338.6 ng.g-1 dw). On the other hand, the least removed compound in both the long-tailed duck and the razorbill was 4-t-OP. The removal of phenol derivatives from claws in both bird species was at the same level. However, 4-NP concentrations were found to be statistically significantly higher in razorbill remiges compared to those of the long-tailed duck (p < 0.05). Comparison of concentrations in the remiges of the long-tailed duck and the razorbill, moulted in two different environments with different levels of pollution and distances from sources, indicated that the Baltic Sea is approximately 3 times more polluted with 4-NP than the marine areas of the Russian Arctic. This demonstrates the potential for the use of 4-NP and remiges as indicators of environmental pollution with phenol derivatives.

Risk to Public Health Regarding Doxycycline Residues in Chicken Claws.

Doxycycline residues in claws may pose a potential risk to humans. A study with broiler chickens was performed to find the concentrations of doxycycline in claws after treatment. For the quantification of the antibiotic of interest in claws, an ultrahigh-performance liquid chromatography-tandem mass spectrometry method was developed. Doxycycline was detectable at 3043 µg/kg at sub-therapeutic treatment, 223 µg/kg after therapeutic doses, and 14 µg/kg after spray application on the last day of the experiment (day 22). Almost 70 days is required for the concentration of doxycycline to reach zero. The obtained results show that monitoring of doxycycline in claws should be implemented.

Mineral composition and microstructure of the abaxial hoof wall in dairy heifers after biotin supplementation.

The aim of this study was to evaluate the effect of daily biotin supplementation on the mineral composition and microstructure of the abaxial hoof wall in dairy heifers. The heifers were housed on a concrete floor and fed for weight gain more than 800 g per day, which is a challenging environment for the hoof. Twelve crossbred dairy heifers (Jersey × Holstein) were divided into two treatment groups. Animals in the control group (n = 6) received a diet without supplemental biotin, while the heifers in the biotin-supplemented feed group (n = 6) each received 20 mg of biotin daily for 120 days. Samples of the abaxial hoof wall were collected from the outer claw of the fore and hind limb, before and after supplementation. The samples were evaluated by X-ray fluorescence spectrometry, computed microtomography, atomic force microscopy and confocal laser scanning microscopy. Biotin supplementation increased the sulphur content and decreased the calcium and potassium content in the abaxial hoof wall. Biotin treatment also increased the percentage of horn tubules with smaller diameter marrow (17-51 µm). However, biotin did not influence the surface relief of the hoof wall, suggesting that its action is limited to the inner layers of the stratum corneum. Daily supplementation with 20 mg of biotin promoted changes in the mineral composition and microstructure of abaxial hoof wall of crossbred dairy heifers. These findings suggest biotin supplementation improves hoof quality and may help to understand the function of biotin in the stratum corneum.

Immunolocalization of corneous beta proteins of the Epidermal Differentiation Complex in the developing claw of the alligator.

Knowledge on the sharpness, mechanical and hydration resistance of the corneous material of claws requires information on its constituent proteins. The present immunohistochemical study has localized some of the main corneous beta proteins (CBPs, formerly termed beta-keratins) indicated to be present in alligator claws only by genomic data. Using specific antibodies we show the immunolocalization of representative claws CBPs of the Epidermal Differentiation Complex (Beta A1 group) during late stages of claw development in alligator. Intense but asymmetric proliferation, revealed by 5BrdU-immunolabeling, determines the formation of a curved dorsal part (unguis) and a linear ventral part (sub-unguis). The large beta-cells generated in the unguis and their packing into a solid corneous layer occur before thinner beta-cells appear in the sub-unguis. In the latter, CBPs are also immune-detected but with less intensity compared to the unguis, and corneocytes remain separated and desquamate. It is suggested that at the tip of the developing claw beta-corneocytes move downward into the initial part of the sub-unguis. This circular movement contributes to sharpen the claw as these cells fully cornify and are desquamated from the sub-unguis. Corneocytes of the unguis contain 10-16 kDa proline-serine-rich proteins that also possess high percentages of glycine, cysteine, tyrosine, valine and leucine. Cysteines likely give rise to numerous SS bonds in the constituent hard horny material, tyrosine contribute to packing proteins into a dense horny material while glycine, valine and leucine increase the hydrophobic property of claws in these water-adapted predators.

Corneous beta proteins of the epidermal differentiation complex (EDC) form large part of the corneous material of claws and rhamphothecae in turtles.

The presence of specific protein types in claws and beaks of turtles is poorly known. The present immunological study describes the localization of some of the main corneous beta proteins (CBPs) coded in the epidermal differentiation complex of turtles. Three antibodies here utilized revealed that glycine-, cysteine-, tyrosine-, and valine-rich CBPs are present in differentiating keratinocytes of the beak and of the dorsal (unguis) and ventral (sub-unguis) sides of the claw in different species, semi-aquatic and terrestrial. These proteins provide mechanical resilience to the horny material of claws and beaks through the formation of numerous -S-S- bonds and also hydrophobicity that contributes to preserve wearing of the horny material. The thicker corneous layer of the unguis is made of elongated and partially merged corneocytes, and no or few cells desquamate superficially. Unknown junctional proteins may contribute to maintain corneocytes connected one to another. In contrast, corneocytes of the sub-unguis show an elongated but lenticular shape and form a looser corneous layer whose cells remain separate and desquamate superficially. This suggests that other specific corneous proteins are present in the unguis in comparison with the sub-unguis to determine this different compaction. The wearing process present in the sub-unguis creates a loss of tissue that may favor the slow by continuous apical migration of corneocytes from the unguis into the initial part of the sub-unguis. Beak corneocytes form a compact corneous layer like the unguis but numerous superficial cells desquamate on both outer (epidermal) and inner (oral) sides.

Perinatal concentrations of 17ß-estradiol and testosterone in the toe claws of female and male dogs from birth until 60 days of age.

This study was conducted to assess concentrations of 17ß-estradiol (E2) and testosterone (T) in toe claws of puppies collected at birth, at 30 and at 60 days of age, evaluating changes relating to age and effect of puppy sex, Apgar score, bodyweight at birth, "litter effect", litter size, and maternal age. Puppies (n = 89), 46 males and 43 females, with normal weight and without malformations, were assigned for the study. Within 12 h of birth tips of toe claws were clipped, and the re-growth tissue of the claws was collected at 30 and 60 days of age. Steroid quantifications occurred using a radioimmunoassay. The results indicated there were lesser concentrations (P < 0.001) of both hormones at 30 and 60 days of age than at birth and that concentrations were similar at 30-60 days of age. There were greater (P < 0.001) T concentrations in males than females, with there being an interaction between sex and sampling time (P < 0.01). The Apgar score was positively (P < 0.001) related to T concentrations in toe claws at birth. The bodyweight was positively correlated (P < 0.05) with T concentrations, with an interaction among puppy sex, bodyweight and sampling time (P < 0.05). Results of the present study confirmed the usefulness of toe claws as a matrix for study of hormonal changes in perinatology of dogs. Results of the study also indicate there are greater E2 and T concentrations at birth compared with 30 and 60 days of age that could be the result of these prenatal steroids affecting fetal development.

Liquefaction of porcine hoof shell to prepare peptone substitute by instant catapult steam explosion.

Instant catapult steam explosion (ICSE) was proposed as a method to liquefy porcine hoof shell (PHS) and prepare a peptone substitute for fermentation culture, achieving environmentally friendly animal by-product recycling. The liquefaction of PHS was conducted at various pressures (0.5-2.3 MPa) for 5-30 min. As evidenced by the scanning electron microscopy analysis, ICSE caused randomly cracks changing the morphological structure of the solid fraction, and ultimately led to protein migration from the solid to liquid phase. Moreover, the chromatographic analysis revealed that the main constituents of the liquid fraction were short peptides (<2 kDa, 84.72%) and amino acids (1.68 mg/mL) at the pressure of 2.3 MPa for 30 min. Subsequently, liquid fractions were prepared as a PHS peptone substitute for fermentation culture. Results suggested the PHS peptone substitute as the main nitrogen source in media was more suitable for the growth of fungus. Therefore, ICSE provides a possibility of large-scale environmentally sustainable management of animal by-products through liquefaction.

Differences in mineral concentrations on hooves of horses, mules and donkeys / Diferenças nas concentrações de minerais nos cascos deequinos, muares e asininos

This study aimed to measure the concentrations of minerals in different regions and pigmentation of horses’ hooves, mules and donkeys. A total of 20 Crioulo horses, 20 mules and 20 Pega donkeys were used. They formed two groups: pigmented (dark) or non-pigmented (bright) hooves. All animals were unshod and with adequate foot conformation. Females were nonpregnant and without foal and no animals were in training. Samples were collected from the hooves of all limbs in the regions of wall, sole and frog. The concentrations of calcium, magnesium, copper, iron and zinc were determined by atomic absorption spectrophotometry; sodium and potassium concentrations were determined by atomic emission spectrophotometry. Among the groups it was observed that the wall, sole and frog of donkeys’ hooves presented higher concentrations of sodium and calcium and with the exception of the frog that also obtained higher levels of copper in this species. Mules had lower concentrations of zinc in the wall and the horses had higher concentrations of potassium and zinc in the sole and higher magnesium in the frog and wall. There was no difference (P>0.05) in concentrations regarding the pigmentation of the hooves. The different regions of the hooves in each group presented similar concentrations of minerals and it was regardless of the pigmentation. The mineral concentrations may be related to desirable functional properties of the donkeys’ hooves. Pigmented and non-pigmented hooves have similar concentrations of minerals.

O objetivo desse trabalho foi mensurar os teores de minerais nas diferentes regiões e pigmentações dos cascos de equinos, muares e asininos. Utilizaram-se 20 equinos da raça Crioula, 20 muares e 20 asininos da raça Pêga. Formaram-se dois grupos conforme os cascos pigmentados (escuros) e não pigmentados (claros). Todos os animais estavam desferrados e com adequada conformação podal e as fêmeas estavam não gestantes e sem potro ao pé e nenhum animal estava em treinamento. Amostras foram coletadas dos cascos de todos os membros nas regiões de parede, sola e ranilha. As concentrações de cálcio, magnésio, cobre, ferro e zinco foram determinadas por espectrofotometria de absorção atômica e as concentrações de sódio e potássio foram determinadas por espectrofotometria de emissão atômica. Entre os grupos observou-se que a parede, sola e ranilha nos asininos apresentaram maiores valores de sódio e cálcio e, exceto para a ranilha, também obteve-se maiores teores de cobre nesta espécie. Os muares apresentaram menores teores de zinco na parede e, os equinos, maiores teores de potássio e zinco na sola e ranilha e de magnésio na ranilha e parede. Não houve diferença (P>0,05) nos teores dos minerais quanto à pigmentação dos cascos. As diferentes regiões dos cascos em cada grupo apresentaram teores similares de minerais e este independeu da pigmentação. As variações dos teores de minerais entre os grupos podem estar relacionados a propriedades funcionais desejáveis dos cascos dos asininos. Cascos pigmentados e não pigmentados apresentam teores similares dos minerais.

Comparison of the peptidome released from keratins in Saiga antelope horn and goat horn under simulated gastrointestinal digestion.

Goat horn (Caprae Hircus Cornu, GH) has been used as a substitute for Saiga antelope horn (Saigae Tataricae Cornu, SAH) in the clinic and the pharmaceutical industry. In the present study, peptides released from SAH and GH under simulated gastric and intestinal digestion were identified. The results showed that most of the peptides released from SAH and GH under simulated gastrointestinal digestion were hydrophilic, and over 75% of the peptides from keratins (KRTs) were hydrophilic. In total, over 58% of the identified peptides were released from KRTs, and were from the four main regions of KRTs. The peptide features and the peptide release profiles from KRTs in SAH and GH were similar, which may provide a method for the identification of sustainable alternatives to replace the threatened SAH, and provide further evidence of the feasibility of using GH as a replacement for SAH based on their peptidomic analysis.

Molecular structure of sauropsid ß-keratins from tuatara (Sphenodon punctatus).

The birds and reptiles, collectively known as the sauropsids, can be subdivided phylogenetically into the archosaurs (birds, crocodiles), the testudines (turtles), the squamates (lizards, snakes) and the rhynchocephalia (tuatara). The structural framework of the epidermal appendages from the sauropsids, which include feathers, claws and scales, has previously been characterised by electron microscopy, infrared spectroscopy and X-ray diffraction analyses, as well as by studies of the amino acid sequences of the constituent ß-keratin proteins (also referred to as the corneous ß-proteins). An important omission in this work, however, was the lack of sequence and structural data relating to the epidermal appendages of the rhynchocephalia (tuatara), one of the two branches of the lepidosaurs. Considerable effort has gone into sequencing the tuatara genome and while this is not yet complete, there are now sufficient sequence data for conclusions to be drawn on the similarity of the ß-keratins from the tuatara to those of other members of the sauropsids. These results, together with a comparison of the X-ray diffraction pattern of tuatara claw with those from seagull feather and goanna claw, confirm that there is a common structural plan in the ß-keratins of all of the sauropsids, and not just those that comprise the archosaurs (birds and crocodiles), the testudines (turtles) and the squamates (lizards and snakes).

A natural energy absorbent polymer composite: The equine hoof wall.

The equine hoof has been considered as an efficient energy absorption layer that protects the skeletal elements from impact when galloping. In the present study, the hierarchical structure of a fresh equine hoof wall and the energy absorption mechanisms are investigated. Tubules are found embedded in the intertubular matrix forming the hoof wall at the microscale. Both tubules and intertubular areas consist of keratin cells, in which keratin crystalline intermediate filaments (IFs) and amorphous keratin fill the cytoskeletons. Cell sizes, shapes and IF fractions are different between tubular and intertubular regions. The structural differences between tubular and intertubular areas are correlated to the mechanical behavior of this material tested in dry, fresh and fully hydrated conditions. The stiffness and hardness in the tubule areas are higher than that in the intertubular areas in the dry and fresh samples when loaded along the hoof wall; however, once the samples are fully hydrated, the intertubular areas become stiffer than the tubular areas due to higher water absorption in these regions. The compression behavior of hoof in different loading speed and directions are also examined, with the isotropy and strain-rate dependence of mechanical properties documented. In the hoof walls, mechanistically the tubules serve as a reinforcement, which act to support the entire wall and prevent catastrophic failure under compression and impact loading. Elastic buckling and cracking of the tubules are observed after compression along the hoof wall, and no shear-banding or severe cracks are found in the intertubular areas even after 60% compression, indicating the highly efficient energy absorption properties, without failure, of the hoof wall structure. STATEMENT OF SIGNIFICANCE: The equine hoof wall is found to be an efficient energy absorbent natural polymer composite. Previous studies showed the microstructure and mechanical properties of the hoof wall in some perspective. However, the hierarchical structure of equine hoof wall from nano- to macro-scale as well as the energy absorption mechanisms at different strain rates and loading orientations remains unclear. The current study provides a thorough characterization of the hierarchical structure as well as the correlation between structure and mechanical behaviors. Energy dissipation mechanisms are also identified. The findings in the current research could provide inspirations on the designs of impact resistant and energy absorbent materials.

The usefulness of claws collected without invasiveness for cortisol and dehydroepiandrosterone (sulfate) monitoring in healthy newborn puppies after birth.

Despite the high rates of perinatal mortality in dogs, the study of canine perinatology did not receive strong scientific interests until recently, probably due to the difficulties in studying long term changes evaluations without incur with ethical issues. In the recent years, however, the usefulness of new matrices, such as the claws, collectable without invasiveness and providing information about a long-term, retrospective window of time were reported to be a suitable tool for the study of canine perinatology in spontaneously dead puppies. The present study was therefore aimed to assess the usefulness of claws as matrices collectable without invasiveness for the study of immunoreactive cortisol and dehydroepiandrosterone (sulfate) in alive puppies at birth, at 30 and at 60 days of age. Beside the effect played by the sampling time, the possible effect of type of birth, vaginal delivery (VD) or elective Cesarean section (CS), as well as of some other maternal or neonatal factors was assessed. The results showed a significant decrease of both hormones from birth to 30 and to 60 days of age (p < 0.001) and from 30 to 60 days of age (p < 0.05), and highlighted, for both hormones, the influence of the type of birth, with newborns born by VD having higher concentrations of immunoreactive cortisol (p < 0.01) and dehydroepiandrosterone (sulfate) (p < 0.001) than the newborns born by elective CS in collections made at birth. No other significant effect was detected. The study confirmed the usefulness of claws as matrix collectable without invasiveness for the retrospective, long-term assessment of hormonal changes in alive newborn puppies and that both hormones declined from birth to 60 days of age. The differences between puppies born by vaginal delivery or elective Caesarean section suggest a possible different HPA activation in puppies born by the two types of birth.

Using Non-destructive Techniques to Measure Mercury (Hg) Concentrations in Gravid Blanding's Turtles (Emydoidea blandingii) in Northeastern Illinois.

Aquatic turtles are suitable biomonitors of wetland ecosystem health because they are long-lived and occupy elevated trophic positions in wetland food webs. This study aimed to determine Hg exposure in adult Blanding's turtles (Emydoidea blandingii), an imperiled prairie-wetland species endemic to the northern U.S. and southern Canada. Claw samples were collected from gravid females from four wetland sites in northeast Illinois. Claw Hg concentrations ranged from 654 to 3132 ng/g and we found no effect of body size (carapace length, CL) and some evidence for an effect of wetland site (WS) on mean Hg (i.e. weak effect of site on Hg, detected between WS1 and WS3). Claw Hg concentrations reported in this study were lower than claw concentrations published for other freshwater turtles (e.g. Chelydra serpentina, Sternotherus oderatus). This is the first Hg-related study on Blanding's turtles and can serve as a reference for other Hg studies in Illinois wetlands.

Trace Element Concentrations in European Pond Turtles (Emys orbicularis) from Brenne Natural Park, France.

We assessed trace elements concentration in European pond turtle (Emys orbicularis) from Brenne Natural Park (France). We sampled road-killed turtles (N = 46) to measure the concentrations of 4 non-essential (Ag, Cd, Hg, and Pb) and 10 essential (As, Co, Cr, Cu, Fe, Mn, Ni, Se, V, and Zn) elements in muscle, skin, liver and claws. Body size or sex did not influence the concentrations of most elements; except for Hg (liver, skin and claws) and Zn (muscle) which increased with body size. We found relatively high concentrations of Hg and Zn, possibly linked to fish farming. This result deserves future investigations to evaluate possible ecotoxicological effects on E. orbicularis.

Development and Validation of an HPLC-UV Method to Quantify Tavaborole During in Vitro Transungual Permeation Studies.

A selective and rapid reversed-phase HPLC-UV method was developed and validated to quantify tavaborole (TAV; AN2690) in biological samples, i.e., in receiving phase and in bovine hoof membrane extract derived from in vitro transungual permeation studies. A simple solid-liquid extraction procedure was used to recover the drug from the bovine hoof slices. TAV chromatographic separation was achieved on a Luna PFP column (150 × 4.6 mm, 5 µm) using a mobile phase consisting of a 70% phosphoric acid solution (10 mM, pH 2.0) with 30% acetonitrile. The detection wavelength was set to 220 nm using a UV detector. The method exhibited good linearity in the calibration ranges, which were 0.5-8.0 and 0.03-2.5 µg/mL for the receiving phase and hoof membranes, respectively. The obtained LOD and LOQ values were 0.023 and 0.069 µg/mL, respectively, for the receiving phase and 0.0024 and 0.007 µg/mL for the bovine hoof membrane extracts. In all cases, the CV for intraday and interday precision was widely below the limit of 2%, demonstrating good precision. The analytical method described was sensitive, precise, linear, and accurate and could be applicable for clinical and bioanalytical studies as an alternative to other analytical methods, which are quite expensive and not always available in research laboratories.

Microtomographic Parameters and Nanoindentation of the Hoof of Girolando Cattle.

The aim of this study was to describe the microstructure of the pigmented and depigmented hoof capsule of Girolando cattle by bi- and tridimensional microtomography and nanoindentation, analysing the possible relation between these findings and the susceptibility of such animals to podal diseases. To carry out the microtomography and the nanoindentation, duplicated samples were collected from the dorsal wall, abaxial wall and pre-bulbar sole of the hoof capsule. Material collection was performed in 40 medial digits of thoracic limbs and 40 lateral digits of pelvic limbs. The bidimensional microtomography showed that the dorsal wall of the thoracic and pelvic limbs presented higher density, followed by the abaxial wall, and finally by the sole, with the lowest density. Moreover, the hoof capsule of cows of Girolando breed is a compact, non-porous material, and constituted by extratubular and intratubular keratin. By the tridimensional microtomography, it was possible to measure the angles of the corneal tubules in relation to the periople and the claws in the different regions of the hoof capsule, which were 90° for the dorsal wall, 55° for the abaxial wall and 70° for the sole. The tridimensional microtomography also showed corneal tubules of different diameters: 17, 51, 85, 119 and 153 µm. The nanoindentation test, when performed in different regions of the hoof capsule, did not reveal significant difference of Vickers hardness in the evaluated areas. However, we verified a larger elastic module of these regions on the transversal cut of the corneal tubules compared to the longitudinal cut.

Biochemical composition of the hoof capsule of buffaloes and its influence on hoof quality / Composição bioquímica do estojo córneo de bubalinos e sua influência na qualidade do casco

The purpose of this study was to establish the biochemical parameters of the abaxial wall, dorsal wall and sole of the hoof of the medial thoracic, lateral, and medial pelvic digits of buffalos. The hoof samples were subjected to destructive biochemical analyses to identify the dry material (DM), mineral matter (MM), organic matter (OM), crude protein (CP) and ether extract (EE) contents. Sulfur (S), calcium (Ca), potassium (K), phosphorus (P), zinc (Zn) and copper (Cu) levels were determined based on nondestructive biochemical analyses. The parameters of dry material, mineral matter, organic matter, crude protein and ether extract of hoof capsule of the digits of buffalos can be determined by means of both destructive and nondestructive biochemical analysis. In addition, this study revealed that the highest concentrations of DM, CP and minerals such as, K, Zn and Cu are concentrated in the digits that bear the greatest body mass weight, suggesting that there is a positive correlation between the aforementioned parameters and the strength and growth of the hoof capsule in the digits. As for the element S, this study demonstrated that its highest concentration is located in the lateral digits of the pelvic members.(AU)

O presente estudo objetivou estabelecer os parâmetros bioquímicos da muralha abaxial, muralha dorsal e sola do casco dos dígitos torácico medial e pélvico lateral dos bubalinos. Foram realizadas analises bioquímicas destrutivas das amostras dos cascos para se obterem os teores de matéria seca (MS), matéria mineral (MM), matéria orgânica (MO), proteína bruta (PB) e extrato etéreo (EE). As análises bioquímicas não destrutivas foram empregadas para se avaliarem os níveis de enxofre (S), cálcio (Ca), potássio (K), fósforo (P), zinco (Zn) e cobre (Cu). Concluiu-se que os parâmetros matéria seca, matéria mineral, matéria orgânica, proteína bruta e extrato etéreo do estojo córneo dos dígitos de bubalinos podem ser definidos por análises bioquímicas destrutivas e não destrutivas. Os maiores valores dos elementos minerais estão concentrados nos dígitos que suportam o maior peso do animal, sugerindo que existe uma relação positiva entre esses parâmetros. Além disso, este estudo revelou que as maiores concentrações de MS, PB e minerais como, K, Zn e Cu estão nos dígitos que carregam o maior peso de massa corporal, o que indica que há uma correlação positiva entre os parâmetros acima referidos, resistência e crescimento do estojo córneo dos dígitos. Em relação ao elemento de S, este estudo demonstrou que a sua maior concentração situa-se nos dígitos laterais dos membros pélvicos.(AU)

Filamentous Structure of Hard ß-Keratins in the Epidermal Appendages of Birds and Reptiles.

The structures of avian and reptilian epidermal appendages, such as feathers, claws and scales, have been modelled using X-ray diffraction and electron microscopy data, combined with sequence analyses. In most cases, a family of closely related molecules makes up the bulk of the appendage, and each of these molecules contains a central ß-rich 34-residue segment, which has been identified as the principal component of the framework of the 3.4 nm diameter filaments. The N- and C-terminal segments form the matrix component of the filament/matrix complex. The 34-residue ß-rich central domains occur in pairs, related by either a parallel dyad or a perpendicular dyad axis, and form a ß-sandwich stabilized by apolar interactions. They are also twisted in a right-handed manner. In feather, the filaments are packed into small sheets and it is possible to determine their likely orientation within the sheets from the low-angle X-ray diffraction data. The physical properties of the various epidermal appendages can be related to the amino acid sequence and composition of defined molecular segments characteristic of the chains concerned.

A novel method for assessing chronic cortisol concentrations in dogs using the nail as a source.

Cortisol, a glucocorticoid secreted in response to stress, is used to assess adrenal function and mental health in clinical settings. Current methods assess cortisol sources that reflect short-term secretion that can vary with current stress state. Here, we present a novel method for the extraction and quantification of cortisol from the dog nail using solid phase extraction coupled to enzyme-linked immunosorbent assay. Validation experiments demonstrated accuracy (r = 0.836, P < 0.001) precision (15.1% coefficients of variation), and repeatability (14.4% coefficients of variation) with this method. Furthermore, nail cortisol concentrations were positively correlated to an established hair cortisol method (r = 0.736, P < 0.001). Nail cortisol concentrations did not differ with dog sex, breed, age, or weights; however, sample size limitations may preclude statistical significance. Nail cortisol may provide information on cortisol secretion integrated over the time corresponding to nail growth and may be useful as a tool for diagnosing stress and adrenal disorders in dogs.

Residue depletion of oxytetracycline (OTC) and 4-epi-oxytetracycline (4-epi-OTC) in broiler chicken's claws by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Antibiotics are widely used in poultry production for the treatment of bacterial diseases. However, residues may remain in products and by-products destined for human consumption or animal feeding. The claws of chickens, which are a by-product of the poultry industry, can directly or indirectly enter the food chain as meals destined to feed other productive animals. Thus, it becomes necessary to determine and quantify antimicrobial residues present in this matrix. The objective of the study was to assess the depletion of oxytetracycline (OTC) and its metabolite 4-epi-OTC in broiler chicken's claws. Claws of 32 broilers treated with a therapeutic dosage of 10% OTC during 7 days were analysed. Samples were taken at days 3, 9, 15 and 19 post-treatment. As for the control group, eight broiler chickens were raised under the same conditions. Extraction was carried out through EDTA-McIlvaine buffer, and clean-up employed a SPE C-18 Sep-Pak®. Instrumental analysis was performed through LC-MS/MS. The concentrations of both analytes were determined in claw samples until day 19 post-treatment. Average concentrations were within the LOD (20 µg kg-1) and LOQ (22 µg kg-1) for OTC and 84 µg kg-1 for 4-epi-OTC. Withdrawal times (WDTs) of 39 days for OTC and 54 days for 4-epi-OTC were established in claws based on 95% confidence. These findings demonstrate that claws can be a source of antimicrobial residue entry into the food chain, since the results showed that OTC and its metabolite can be found in chicken's claws for long periods, even exceeding the average lifespan of a broiler chicken.