Poultry Feathers Research Articles

Development of a keratinase activity assay using recombinant chicken feather keratin substrates.

Poultry feathers consist mainly of the protein keratin, which is rich in β-pleated sheets and consequently resistant to proteolysis. Although many keratinases have been identified, the reasons for their substrate specificity towards β-keratin remain unclear due to difficulties in preparing a soluble feather keratin substrate for use in activity assays. In the present study, we overexpressed Gallus gallus chromosomes 2 and 27 β-keratin-encoding genes in Escherichia coli, purified denatured recombinant proteins by Ni2+ affinity chromatography, and refolded by stepwise dialysis to yield soluble keratins. To assess the keratinolytic activity, we compared the proteolytic activity of crude extracts from the feather- degrading bacterium Fervidobacterium islandicum AW-1 with proteinase K, trypsin, and papain using purified recombinant keratin and casein as substrates. All tested proteases showed strong proteolytic activities for casein, whereas only F. islandicum AW-1 crude extracts and proteinase K exhibited pronounced keratinolytic activity for the recombinant keratin. Moreover, LC-MS/MS analysis of keratin hydrolysates allowed us to predict the P1 sites of keratinolytic enzymes in the F. islandicum AW-1 extracts, thereby qualifying and quantifying the extent of keratinolysis. The soluble keratin-based assay has clear therapeutic and industrial potential for the development of a high-throughput screening system for proteases hydrolyzing disease-related protein aggregates, as well as mechanically resilient keratin-based polymers.

PRODUCTION OF ANTIMICROBIAL AND ANTICANCER FOM FEATHER-KERATINOLYTIC NOCARDIOPSIS SP. 28ROR AS A NOVEL STRAIN USING FEATHER MEAL MEDIUM

Objective: Production of bioactive secondary metabolites from a feather-degrading actinobacterial species using feather meal medium. Methods: Protease producer actinobacterial isolates (22) recovered from farm soil, poultry farm soil and feather wastes were used to test the antagonistic effect on pathogenic bacteria and fungi, including Staphylococcus aureus, Escherichia coli, Microsporum canis and Trichophyton mentagrophyte using Mueller-Hinton agar and potato dextrose agar (PDA) media. The best isolate was used to produce the active metabolites in feather meal medium composed from (g/l) 10 feather meal, 5 sucrose and 0.3 l cement extract were dissolved in tap water, in addition to the standard medium composting of (g/l) glucose (1%), tryptone (1%), KH2PO4 (0.07%) and K2HPO4 (0.14%) were dissolved in distilled water, both of them at initial pH 9. The secondary metabolites were partially purified by Sephadex LH20 column and the antimicrobial activity and cytotoxic activity were assayed.Results: 31.82% of isolates inhibited the growth of both bacterial and fungal test organisms and the best one was Nocardiopsis sp. 28ROR (GenBank: KC702802.1) at a significant level P˂ 0.05. It produced bioactive metabolites in both feather meal broth and the standard media. The partially purified metabolites inhibit the breast cancer cell line MCF-7 (51%) and normal hepatic cell line WRL-68 (9%), in addition to inhibiting S. aureus andTrichophyton menta agrophyte growth.Conclusion: The actinobacteria has vast abilities to degrade very complex wastes and converted to simple constituents to reprocess in other industries. So the Nocardiopsis sp. 28ROR was a novel strain produced anticancer, antimicrobial substances using feather meal medium as a cheap waste medium.

The structure and properties of viscoelastic polyurethane foams with Fyrol and keratin fibers

The subject of the research presented in this article is viscoelastic polyurethane foams (VPF) made using a halogen free flame retardant additive – Fyrol™ – and keratin fibers. The foams were made with varying isocyanate index. For their modification, Fyrol PNX LE and Fyrol PNX were used as well as keratin fibers obtained from poultry feathers. Foams were characterized by means of techniques such as infrared spectroscopy, thermogravimetric analysis and differential scanning calorimetry. The changes in the patterns of the foams were analyzed using a scanning electron microscope. Moreover, flammability of selected foams was defined using a cone calorimeter and oxygen index. Compression set of the foams was assessed. As a result, it was concluded that the use of a Fyrol mixture with keratinous fibers preferably reduces permanent deformation of foams and a fire hazard caused by their usage.

Synthesis and Characterization of Photo-Cross-Linkable Keratin Hydrogels for Stem Cell Encapsulation

The objective of this work was to synthesize an injectable and photopolymerizable hydrogel based on keratin extracted from poultry feather for encapsulation and delivery of stem cells in tissue regeneration. Since feather keratin is rich in cysteine residue, allylation of sulfhydryl groups was used for functionalization of keratin. Keratin was extracted from feather barbs by reducing the disulfide bonds in cysteine residues to sulfhydryl groups (-SH). Next, the free thiol groups were converted to dehydroalanine (Dha) by oxidative elimination using O-(2,4,6-trimethylbenzenesulfonyl) hydroxylamine. Then, the Dha moieties were converted to s-allyl cysteine by reaction with allyl mercaptan to produce keratin allyl thioether (KeratATE) biopolymer. Human mesenchymal stem cell (hMSCs) were suspended in the aqueous solution of KeratATE, injected into a mold, and photopolymerized to generate a KeratATE hydrogel encapsulating hMSCs. The freeze-dried photo-cross-linked KeratATE hydrogels had a porous, interconnected, honeycomb microstructure with pore sizes in the 20-60 μm range. The compressive modulus of the hydrogels ranged from 1 to 8 kPa depending on KeratATE concentration. KeratATE hydrogels had <5% mass loss in collagenase solution after 21 days of incubation, whereas the mass loss was 15% in trypsin solution. Degradation of KeratATE hydrogel was strongly dependent on trypsin concentration but independent of collagenase. hMSCs proliferated and adopted an elongated spindle-shape morphology after seeding on KeratATE hydrogel. KeratATE hydrogel supported differentiation of the encapsulated hMSCs to the osteogenic and chondrogenic lineages to the same extent as those hMSCs encapsulated in gelatin methacryloyl hydrogel. The results suggest that keratin allyl thioether hydrogel with controllable degradation is a viable matrix for encapsulation and delivery of stem cells in tissue regeneration.

Assessment of the effect of housing on feather damage in laying hens using IR thermography

Plumage damage represents one of the animal-based measures of laying hens welfare. Damage occurs predominantly due to age, environment and damaging pecking. IR thermography, due to its non-invasiveness, objectivity and repeatability is a promising alternative to feather damage scoring systems such as the system included in the Welfare QualityR assessment protocol for poultry. The aim of this study was to apply IR thermography for the assessment of feather damage in laying hens kept in two housing systems and to compare the results with feather scoring. At the start of the experiment, 16-week-old laying hens (n=30) were divided into two treatments such as deep litter pen and enriched cage. During 4 months, feather damage was assessed regularly in 2-week intervals. One more single assessment was done nine and a half months after the start of the experiment. The feather damage on four body regions was assessed by scoring and IR thermography: head and neck, back and rump, belly, and underneck and breast. Two variables obtained by IR thermography were used: the difference between the body surface temperature and ambient temperature (ΔTB) and the proportion of featherless areas, which were defined as areas with a temperature >33.5°C. Data were analyzed using a GLM model. The effects of housing, time, region and their interactions on feather damage, measured by the feather scoring and by both IR thermography measures, were all significant (P<0.001). The ΔTB in all assessed regions correlated positively with the feather score. Feather scoring revealed higher damage in enriched cages compared with deep litter pens starting from week 6 of the experiment on the belly and back and rump regions, whereas ΔTB from week 6 in the belly and from week 8 on the back and rump region. The proportion of featherless areas in the belly region differed significantly between the housings from week 8 of the experiment and on the back and rump region from week 12. The IR thermography assessment of the feather damage revealed differences between hens kept in different housing systems in agreement with the feather scoring. In conclusion, it was demonstrated that the IR thermography is a useful tool for the assessment of poultry feather cover quality that is not biased by the subjective component and provides higher precision than feather damage scoring.

Biodegradation of feather waste keratin by a keratinolytic soil fungus of the genus Chrysosporium and statistical optimization of feather mass loss

This paper assesses the ability of strains of Aphanoascus fulvescens and Chrysosporium articulatum isolated from soil (phaesol) to degrade native feather keratin. Strains were identified based on phenotypic traits and nucleotide sequencing. Response Surface Methodology was used to optimize cultivation conditions exhibiting the highest keratinolytic activity. The experiments were based on Box-Behnken designs for the loss of substrate mass (chicken feathers). While substrate mass loss is an “economic coefficient” that reliably indicates feather keratin degradation, it has not been studied before. Stationary liquid cultures of five selected strains were conducted in laboratory conditions at 28 °C using poultry feathers (1 g) as the sole source of carbon, nitrogen and energy. Enzymatic activities, keratin mineralization products and substrate mass loss were determined periodically. The mineralization of keratin proteins by strains yielded a high number of ammonium ions alkalinizing the medium. Increased ammonium ions inhibited the activity of caseinian protease and keratinase. A decrease in the concentration of these ions induced proteolytic enzymes, chiefly the activity of keratinase, at the end of fungal cultivation. Keratinase activity was related to protein- and peptide release and that of caseinian protease to sulfate ions. The highest loss of substrate mass in comparison to the reference strain CBS104.62 (35.4%) was recorded for Aphanoascus fulvescens B21/4-5 (65.9%). Based on a Box-Behnken design, the maximum loss of substrate mass for the Aphanoascus fulvescens strain (71.08%) can be achieved at pH 7.58 and temperature 28.7 °C.

A randomized, double-blinded crossover trial testing the benefit of two hydrolysed poultry-based commercial diets for dogs with spontaneous pruritic chicken allergy.

Hydrolysed protein diets are used to diagnose and treat dogs with cutaneous adverse food reactions (CAFR). Little is known about what proportion of dogs hypersensitive to the native protein would react to its hydrolysed form. To determine the clinical allergenicity of hydrolysed poultry feather (RCU) and chicken liver diets (HZD) in dogs with chicken induced CAFR. In this randomized, double-blinded, crossover trial, ten dogs with chicken induced CAFR were selected after a positive oral challenge to chicken meat and a negative one to corn. Test diets were fed for 14days separated by a 14 day wash-out period. Owners rated pruritus daily with a Visual Analog Scale (PVAS). The challenge was ended if a flare in pruritus occurred (i.e. PVAS ≥5/10). The median PVAS scores before feeding RCU and HZD were 0.9 and 1.7, respectively (Wilcoxon signed rank test, P=0.46). Pruritus scores increased significantly after feeding HZD (Friedman's test, P<0.001) but not after feeding RCU (P=0.895). None of the dogs fed RCU, but four dogs fed HZD (40%), were withdrawn after a flare in pruritus developed (Fisher's test, P=0.04). The maximal PVAS score was significantly higher after HZD (median: 4.7) compared to RCU (2.5) (Wilcoxon signed rank test, P=0.01). One dog in each group was withdrawn due to diarrhoea. The hydrolysed poultry feather diet did not induce pruritus flares in dogs allergic to chicken in contrast to the hydrolysed chicken liver diet that led to pruritus flares in 40% of these dogs.

Cysteic Acid in Dietary Keratin is Metabolized to Glutathione and Liver Taurine in a Rat Model of Human Digestion.

Poultry feathers, consisting largely of keratin, are a low-value product of the poultry industry. The safety and digestibility of a dietary protein produced from keratin (KER) was compared to a cysteine-supplemented casein-based diet in a growing rat model for four weeks. KER proved to be an effective substitute for casein at 50% of the total dietary protein, with no changes in the rats’ food intake, weight gain, organ weight, bone mineral density, white blood cell counts, liver glutathione, or blood glutathione. Inclusion of KER in the diet reduced total protein digestibility from 94% to 86% but significantly increased total dietary cysteine uptake and subsequent liver taurine levels. The KER diet also significantly increased caecum weight and significantly decreased fat digestibility, resulting in a lower proportion of body fat, and induced a significant increase in blood haemoglobin. KER is therefore a safe and suitable protein substitute for casein, and the cysteic acid in keratin is metabolised to maintain normal liver and blood glutathione levels.

Evaluation with scanning electron microscopy of Cd, Cu, and Zn removal from aqueous solutions by ash from gasification of poultry feathers

Evaluation with scanning electron microscopy of Cd, Cu, and Zn removal from aqueous solutions by ash from gasification of poultry feathers

Evaluation with scanning electron microscopy of Cd, Cu, and Zn removal from aqueous solutions by ash from gasification of poultry feathers

Evaluation with scanning electron microscopy of Cd, Cu, and Zn removal from aqueous solutions by ash from gasification of poultry feathers

Poultry feather wastes recycling possibility as soil nutrient

Poultry feathers are produced in large amounts as a waste in poultry slaughterhouses. Only 60-70% of the poultry slaughterhouse products are edible for human being. This means more million tons annually worldwide (Papadopoulus et al., 1986; Williams et al., 1991; Hegedűs et al., 1998). The keratin-content of feather can be difficulty digested, so physical, chemical and/or biological pre-treatment are needed in practice, which have to be set according to the utilization method. Feather was enzymatic degraded, and then fermented in separated bioreactors. The anaerobic bioreactor system (4 digesters with 6 litre volume) was controlled by ACE SCADA software running on Linux platforms. Pot scale seed germination tests were established to suggest the quantity of digested slurry to be utilized. The chosen test plants were lettuce (Lactuca sativa) . In case of reproduction test Student’s t-test was applied to examine significant differences between the root lengths of the control and the treated plant species. In case of pot seed germination variance analysis with Tukey B’s and Duncan test was applied to examine significant differences between the root lengths of plants, grown on different treatments. The effect of treatments on germination ability of the plant species was expressed in the percentage of the controls. According to Student’s t-test significant difference was found between root lengths of different treatments. Based on variance analysis with Tukey B’s and Duncan tests could be detected a significant difference between the treatments. Utilization of the fermented material reduces the use of fertilizers and because of its large moisture content it reduces the watering costs. Recycle of the slaughterhouse feather and different agricultural wastes and by-products can solve three main problems: disposal of harmful materials, producing of renewable energy and soil nutrient, measuring reflectance at the certain spectral range, which can facilitate real time water status assessment of orchards.

Isolation, Identification, and Characterization of a Keratin-degrading Bacterium Chryseobacterium sp. P1-3

In this study, a keratin-degrading bacterium was isolated from soil contaminated with feather waste. The isolated strain was identified as Chryseobacterium sp. P1-3 on the basis of the 16S rRNA gene sequence alignment. Chryseobacterium sp. P1-3 is currently used in various biotechnological applications (e.g., in the hydrolysis of poultry feathers). It hydrolyzed the feather meal within 2 days and possesses a high level of keratinase activity (98 U/mL). The keratinase, partially purified from this strain, prefers casein as a substrate and shows optimal activity at a temperature of 30°C and at a pH of 8.0.

Keratinolytic abilities of Micrococcus luteus from poultry waste.

Keratinolytic microorganisms have become the subject of scientific interest due to their ability to biosynthesize specific keratinases and their prospective application in keratinic waste management. Among several bacterial classes, actinobacteria remain one of the most important sources of keratin-degrading strains, however members of the Micrococcaceae family are rarely scrutinized in regard to their applicatory keratinolytic potential. The tested Micrococcus sp. B1pz isolate from poultry feather waste was identified as M. luteus. The strain, grown in the medium with 1–2% chicken feathers and a yeast extract supplement, produced keratinases of 32 KU and lower level of proteases, 6 PU. It was capable to effectively decompose feathers or “soft” keratin of stratum corneum, in contrast to other “hard” hair-type keratins. The produced keratinolytic enzymes were mainly a combination of alkaline serine or thiol proteases, active at the optimum pH 9.4, 55 °C. Four main protease fractions of 62, 185, 139 and 229 kDa were identified in the crude culture fluid. The research on the auxiliary role of reducing factors revealed that reducing sulfur compounds could be applied in keratinolysis enhancement during enzymatic digestion of keratin, rather than in culture conditions. The presented M. luteus isolate exhibits a significant keratinolytic potential, which determines its feasible applicatory capacity towards biodegradation of poultry by-products or formulation of keratin-based feed components.

Combining polyhydroxyalkanoates with nanokeratin to develop novel biopackaging structures

ABSTRACTThis work reports on the combination of polyhydroxyalkanoate (PHA)‐based materials and nanokeratin extracted from poultry feathers (a food by‐product) using different approaches. In the first approach, high‐barrier nanobiocomposites based on the combination of PHAs with a nanokeratin additive were successfully developed via both (i) direct melt compounding and (ii) preincorporated into an electrospun masterbatch of PHA which was subsequently melt compounded with PHA pellets. Enhanced barrier properties for the nanobiocomposites were obtained which were seen to depend on the PHA grade used. In the second approach, nanokeratin films were obtained via solution casting which were successfully hydrophobized by coating them with electrospun PHA fibers. The multilayered approach showed good adhesion and also lead to enhanced barrier performance. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016, 133, 42695.

Oxygen reduction reaction activity of nitrogen, oxygen and sulphur containing carbon derived from low-temperature pyrolysis of poultry featherfiber

Poultry featherfiber (PFF), the soft fibrous part of poultry feathers, is used as the novel precursor to derive N-, O- and S-doped-carbon (CNx) electrocatalysts. The catalysts are synthesized by a low-temperature pyrolysis of PFF or a mixture of PFF and Fe/Co salts in varying concentrations, followed by acid-treatment to remove the metal from pyrolyzed samples. Electrochemical measurements in alkaline media exhibit significant ORR electrocatalytic activity of CNx with an ORR onset potential of ∼-230mV (vs. Ag/AgCl), which is comparable to Pt (∼-210mV) under similar conditions. Despite the absence of metal-carbon bonds, the CNx electrocatalysts synthesized in presence of Fe/Co exhibit catalytic activity significantly higher than that of the CNx synthesized without metals. This is attributed to the modified chemical states of C-N and C-O sites due to the presence of Fe/Co during pyrolysis. Hence, presence of metal precursors during pyrolysis affects the ORR activity of CNx indirectly. The CNx catalysts synthesized in presence of varying Fe to Co ratio contain varying N-content and the ORR peak current is found to vary linearly with the N-content. Observed ORR activity of the present catalyst makes it a suitable Pt-free catalyst for low temperature fuel cells.

Total Elimination of Polluting Chrome Shavings, Chrome, and Dye Exhaust Liquors of Tannery by a Method Using Keratin Hydrolysate

The solid and liquid waste management of tanneries has become a serious problem in the leather industry. In this study, we have attempted and succeeded in eliminating these pollutants by a novel method that has not been done anywhere so far. Keratin hydrolysate (KH) from poultry feathers has been utilized along with chrome shavings (CS) for total elimination of the polluting solid and liquid wastes in a tannery. In this process, 100% fixation of the Cr from the chrome exhaust liquor and the dye from the dye exhaust liquor is achieved by the reactions of CS with KH. The infrared spectra of the different stages of CS confirm the fixation of Cr in the chrome shavings–keratin hydrolysate (CSKH) complex. In this work, apart from elimination of tannery pollutants, a leather-like flexible sheet is obtained that can be used in the footwear and leather goods industry. In the scanning electrom microscopy evaluation, the surface morphology of the product exhibits smoothness because of the incorporation of poly(vinyl alcohol) and ethylene glycol. The above findings will open new avenues for further research in solid and liquid waste management of tannery effluents.

Isolation and Screening of Keratinolytic Bacteria from Feather Dumping Soil in and Around Cuddalore and Villupuram, Tamil Nadu

The study was carried out to screen the keratinolytic bacteria isolated from poultry feather dumping soils collected from Cuddalore and Villupuram in Tamilnadu, India. Forty one isolates were screened from 15 different samples by their growth and feather degrading ability on feather meal agar, and they were named as FDS01 to FDS41. Proteolytic activity of the isolates was screened on casein agar and their efficiency ranged from 4 to 18 mm. Secondary screening of the isolates was determined by keratinase activity. The isolates such as FDS03, FDS05, FDS11, FDS15, FDS17, FDS27 and FDS37 showed maximum keratinase activity and completely decomposed feather in 21 days of incubation (37 °C). In addition, the keratin azure assay was performed with the supernatants in 72 h culture, the isolate FDS15 showed high keratinase specific activity (16.5 ± 2.5 U/ml). The efficient strain was further characterized by biochemical and physiological properties and it was confirmed as Bacillus subtilis by 16S rDNA sequencing.

Wplyw sposobu przygotowania wlokien keratynowych z pior drobiowych na wlasciwosci kompozytow z recyklatow polietylenu duzej gestosci

Composites based on high density polyethylene (PE-HD) matrix with a natural filler in the form of keratin fibers obtained from poultry feather waste were used in the study. Composites containing 15, 30 and 50 % keratin fibers were produced. The fibers were purified using three methods. The water absorption of the obtained materials is dependent on the filler preparation method and is relatively low in comparison with composites containing other natural fillers. Since all of the composites are classified in HB40 flammability category, they belong to semi-flammable materials and therefore they can be used as construction elements of common use with increased resistance to fire. The use of different methods of the purification of poultry feathers in manufacturing process of keratin fibres has not significant influence on the linear shrinkage, density and mechanical strength of the composites containing them.

Microwave-enhanced hydrolysis of poultry feather to produce amino acid

Poultry feather was hydrolyzed at relatively mild high temperature ranging from 433 to 473K and autogenous pressure by intensification of microwave heating. The hydrolysate mainly contains arginine, alanine, threonine, glycine, praline, serine, glutamic acid, aspartic acid, cystine and tyrosine, which corresponds with hydrochloric acid catalytic hydrolysis. Based on the orthogonal experimental result, the total yield of amino acid attains about 54.72% with feather containing about 71.83% keratin at optimum reaction condition of temperature 473K, time 20min and weight ratio of water/feather 37.5. The high yield of amino acid and high efficiency of hydrolysis indicate that the microwave has better intensification on hydrolysis comparing with traditional strong acid catalytic or sub-critical hydrolysis. The apparent activation energies (Ea) are 85.12 and 63.00kJ/mol as to the hydrolysis of feather and the degradation of produced amino acid with consecutive pseudo-first-order reaction kinetic model, and the great decrease of the values comparing with non-microwave heating should be the reason of the enhanced effect of microwave.

Bacteriological and chemical properties of soil amended with fermented poultry bird feather

Huge amount of poultry wastes are generated annually in developing nations, of which end up in our environment. Feather as a poultry waste have caused environmental concerns in many of these countries. In this research work, poultry feather was used to study its effects on bacteriological and chemical properties in agricultural soil. The feathers collected were grinded, sterilized and fermented before mixing it with soil. Total heterotrophic counts of colonies were enumerated from the soil samples. Isolates were characterized based on morphological and series of biochemical tests. Soil chemical analysis for pH, carbon, nitrogen, phosphorus, calcium, potassium was also carried out. The heterotrophic bacteria count of the treated soil had the highest count in the range of 1.54 x106 to 2.02 x106 (CFU/g) as against the control soil with 1.24 x106 (CFU/g). The bacteria identified from these samples were Bacillus sp., Proteus sp., Staphylococcus sp. and Actinomyces sp. Actinomyces sp. was isolated only after three weeks of treatment with the feather while all other bacteria were also present in the control soil but at elevated levels based on heterotrophic bacteria count in the treated soil. The soil chemical analysis of the treated soil had high amount of mineral (Organic carbon 1.10 %, nitrogen 0.09%, phosphorus 0.53±0.02 mg/kg, magnesium 0.45±0.01 mg/kg as against the control (Organic carbon 0.26%, nitrogen 0.05%, phosphorus 0.30±0.10 mg/kg, magnesium 0.45±0.01 mg/kg) and their pH is almost near neutrality, 7.12 and 7.08 respectively. The result suggested that soil treated with poultry feather increased the plate count of Bacillus sp, Proteus sp, Staphylococcus sp and Actinomyces sp. Poultry feather which is a waste by-product can provide essential nutrients for the survival of soil microorganisms, plant growth and therefore can be exploited as biofertilizer.Keywords: Bacillus sp., biofertilizer, fermentation, keratin, poultry waste