Peptide recovery from chicken feather keratin and their anti-biofilm properties against methicillin-resistant Staphylococcus aureus (MRSA).

Bacteria have the potential to adhere to abiotic surfaces, which has an undesirable effect in the food industry because they can survive for sustained periods through biofilm formation. In this study, an antibacterial peptide (ABP), with a molecular mass of 3861 Da, was purified from hydrolyzed chicken feathers using a locally isolated keratinolytic bacterium, namely Rhodococcus erythropolis, and its antibacterial and antibiofilm potential were investigated against planktonic and biofilm cells of Methicillin-Resistant Staphylococcus Aureus (MRSA). The results demonstrated that purified ABP showed the growth inhibition of MRSA cells with the minimum inhibitory concentration (MIC) of 45 µg/ml and disrupted MRSA biofilm formation at a concentration of 200 ug/ml, which results were confirmed by scanning electron micrograph (SEM). Moreover, the secondary structures of the peptide were assessed as part of the FTIR analysis to evaluate its mode of action. ExPASy tools were used to predict the ABP sequence, EPCVQUQDSRVVIQPSPVVVVTLPGPILSSFPQNTA, from a chicken feather keratin sequence database following in silico digestion by trypsin. Also, ABP had 54.29% hydrophobic amino acids, potentially contributing to its antimicrobial activity. The findings of toxicity prediction of the peptide by the ToxinPred tool revealed that ABP had non-toxic effects. Thus, these results support the potential of this peptide to be used as an antimicrobial agent for the treatment or prevention of MRSA biofilm formation in feed, food, or pharmaceutical applications.

The effects of probiotic and threonine application on the carcass yield, internal organ development, intestinal morphology and cecal microbiota of broilers challenged with Clostridium perfringens.

The aim of this study was to explore the effects of probiotics (Ecobiol®) and threonine supplements on broiler internal organs and intestinal health under Clostridium perfringens challenge. A total of 1600 male Ross 308 broiler chicks were randomly assigned to eight treatments with eight replicates each of 25 birds. Dietary treatments consisted of two levels of supplemented threonine (without and with threonine supplementation), two levels of probiotics (Ecobiol®) supplement (0 and 0. 1% of diet), and two levels of challenge (without and with 1 ml of the C. perfringens inoculum (∼108 cfu/ml) on d 14, 15, and 16 of the experiment), which fed to the birds during a 42 d feeding trial. The results showed that adding threonine and probiotic supplements to the diets of C. perfringens-infected birds reduced the relative gizzard weight by 22.9% compared to those fed un-supplemented diet (P ≤ 0.024). As compared to the non-challenged group, the C. perfringens challenge significantly reduced the carcass yield of broilers by 1.18% (P < 0.0004). The groups receiving threonine and probiotic supplementation had higher carcass yield, and the inclusion of probiotics in the diet decreased abdominal fat by 16.18% compared with the control treatment (P ≤ 0.001). Adding threonine and probiotic supplements to the diets of broilers challenged with C. perfringens increased the jejunum villus height in comparison with C. perfringens-infected group fed an unsupplemented diet on day 18 (P ≤ 0.019). The number of cecal E. coli increased in birds under C. perfringens challenge in comparison with the negative group. Based on the findings, dietary inclusion of threonine and probiotic supplement could beneficially affect intestine health and carcass weight during the C. perfringens challenge.

Effects of bioactive peptides derived from feather keratin on plasma cholesterol level, lipid oxidation of meat, and performance of broiler chicks.

In this research, the effect of mixed feather bioactive peptides (MFBPs) added in water, on intestinal health, meat quality, and plasma cholesterol level of broiler chickens, was evaluated. A total of 80 day-old male broiler chicks (Ross 308) were randomly divided into two treatments with four replication pens. The dietary treatments were the drinking water with no additives (control) and drinking water containing 50 mg/L of MFBPs. Live weight and feed intake were measured at the end of starter (1-10 days), grower (11-24 days), and finisher (25-36 days) periods by calculating the average daily gain and feed conversion ratio. The results indicate that body weight gain was greater (P < 0.05) in birds that received MFBPs in the final period. At 24 days of age, the villus height and muscle layer thickness in different parts of the intestine were higher in birds that received bioactive peptides but epithelial thickness was lower than that in control birds (P < 0.05). In addition, the administration of MFBPs decreased (P < 0.01) serum total cholesterol, triglyceride, and low-density lipoprotein in broilers. Supplementation with MFBPs significantly reduced (P < 0.01) the malondialdehyde (MDA) amount in the thigh muscle. In conclusion, using the MFBPs in the diet of broilers could improve meat quality, cholesterol concentration in serum, and gut health.

Stability and cytotoxicity of DPPH inhibitory peptides derived from biodegradation of chicken feather.

The antioxidant activity and cell viability of feather hydrolysates obtained with the Bacillus licheniformis were evaluated using an in-vitro model. The results indicate that feathers-derived peptides under 3 kDa have antioxidant activity with IC50 values of 5.03 ± 0.215 mg/mL by using DPPH antioxidant assay. Although the antioxidant activity of the peptides under 3 kDa preserved after applying diverse heating (from 20 to 100 °C), they lost their activity under strongly acidic or alkaline conditions. Antioxidant activity of the mixed feather bioactive peptides (MFBPs) obtained with partial purification of peptides under 3 kDa was with IC50 amount of 0.169 mg/mL ± 0.004 using DPPH radical scavenging assay. Also, MFBPs within an amount range of from 0.0048 to 5.0 mg/mL, illustrated no cytotoxicity to gingival fibroblast blood cell lines. In light of our results, the obtained value-added peptides could be useful in different food products as a future functional ingredient with antioxidant potency.

Methods of keratin extraction from poultry feathers and their effects on antioxidant activity of extracted keratin.

The antioxidant activities of extracted keratin from chicken feathers using different chemical treatments including 2-mercaptoethanol (TME), sodium sulfite (SS), and sodium dodecyl sulfate (SDS) were investigated using an in-vitro study. The results displayed that all of these methods have antioxidant potency. However, the reduction of keratin with SS displayed the best antioxidant potency with IC50 value of 0.533 ± 0.061 mg/mL using DPPH radical scavenging assay with a porous surface morphology as monitored by scanning electron microscopy (SEM). Compared to raw feathers, all extracted keratins had lower decomposition temperatures and melting points investigated by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) respectively. Fourier-transform infrared spectroscopy (FTIR) confirmed that the extracted keratins maintained the most amides bonds in the procedure of alkaline extraction.

The effect of lactobacillus cell size on its probiotic characteristics.

Screening for probiotic characteristics is usually associated with a series of assays and a large number of isolates to be tested, which can be sometimes costly and frustrating. For this reason, finding some indicators to predict the probiotic potential would be of great significance. In this study, 10 Lactobacillus strains including L. sakei, L. reuteri, L. delbrueckii subsp. lactis, L. delbrueckii subsp. delbrueckii, L. plantarum, L. acidophilus, L. rhamnosus, L. paracasei, L. salivarius, and L. gasseri were characterized by cell morphology and growth properties. The strains were then examined in terms of some probiotic characteristics including resistance to acid and bile conditions, ability to adhesion to intestinal epithelial cells, antioxidant activity, aggregation characteristics, antibacterial activity, hemolytic activity, and resistance to different antibiotics. Correlations between different quantitative features were analyzed using Pearson's coefficient (r). Results of this study provided first-time evidence for the effects of cell length on probiotic features. Based on statistical analysis, long Lactobacillus strains had often higher antioxidant and aggregation activities. Moreover, these long strains were usually more sensitive to acid and bile conditions and resulted in a lower CFU yield compared to short strains. By conducting morphological tests at the first step of screening, some strains would gain higher priority because of predicting a high performance in some of the desired characteristics. Therefore, the cost and time required for the subsequent tests would be significantly reduced.