RESUMO
Feathers make up 7% of the total weight of adult chickens and keratin protein makes up 85% of the feathers. Today, the keratinase enzymes of some Bacillus strains are used to degrade and process raw keratin waste for animal and poultry feed. According to various studies, the probiotic properties of some spore-shaped Bacillus have also been proven. The study aimed to isolation of the keratinolytic Bacillus bacteria that they have probiotic properties for using in the livestock and poultry feed industry. We were able to isolate 8 strains of Bacillus licheniformis with kreatin degrading properties from the soil of Baharan chicken slaughterhouse (Qom city, Iran) applying heat shock, alcohol- and keratin-rich culture medium, and after microscopic and biochemical analysis, 16S rDNA gene was isolated. The measurement results of keratinase activity showed that the three strains of Bacillus licheniformis pvkr6, pvkr 15, and pvkr41 had the highest activity with 124.08, 101.1, and 100.18 U/ml. The results of probiotic properties evaluation also revealed that among all the isolates, only Bacillus licheniformis pvkr15 and Bacillus licheniformis PTCC 1595 (positive control) were γ-hemolytic strains. The percentage of surface hydrophobicity of the strains was obtained from 3.27 to 30.57. It was also shown that, on average, all the strains had acceptable susceptibility to the tested antibiotics except penicillin G. Bacillus licheniformis pvkr15 with highest keratinase activity (101.1U/ml) was considered an optional probiotics due to its abilities such as (biofilm formation, being safe cause of γ-hemolytic activity, high susceptibility to antibiotics such as streptomycin, gentamicin, cefixime, amoxicillin, tetracycline, vancomycin, erythromycin and having a moderate hydrophilic (hydrophobicity: 19.09%), high survivability in pH 2, 2.5 and 3, strong resistance to bile salts and moderate antagonistic activity against pathogenic bacterium like Proteus mirabilis and the ability to grow under anaerobic conditions). By using this strain, after hydrolysis of keratin protein in the feather structure, to replace part of the protein of livestock and poultry feed, not only is no need to separate bacteria from the feed, but also the strain play role of an useful and effective additive in animal growth.
Assuntos
Bacillus , Probióticos , Matadouros , Animais , Antibacterianos/farmacologia , Bacillus/genética , Bacillus/metabolismo , Galinhas , Plumas/química , Plumas/metabolismo , Plumas/microbiologia , Concentração de Íons de Hidrogênio , Queratinas/análise , Queratinas/química , Queratinas/metabolismo , Aves Domésticas/metabolismo , Probióticos/análise , Probióticos/farmacologia , SoloRESUMO
Poultry industry is amongst highly developed industries of Pakistan, fulfilling the protein demand of rapidly increasing population. On the other hand, the untreated poultry waste is causing several health and environmental problems. The current study was designed to check the potential of keratinolytic fungal species for the conversion of chicken-feather waste into biofortified compost. For the purpose, three fungal species were isolated from soil samples. These strains were pure cultured and then characterized phenotypically and genotypically. BLAST searches of 18S rDNA nucleotide sequence of the fungal isolates revealed that the two fungal isolates belonged to genus Aspergillus and one belonged to genus Chrysosporium. Optimum temperature for Aspergillus flavus, Aspergillus niger and Chrysosporium queenslandicum was 29, 26 and 25 oC, respectively. A. flavus showed maximum (53%) feather degradation, A. niger degraded feather waste up to 37%, while C. queenslandicum showed 21% keratinolytic activity on chicken feathers at their respective temperature optima. The degradation potential of these fungal species showed their ability to form compost that has agro-industrial importance.
Assuntos
Compostagem , Plumas , Animais , Galinhas , Plumas/metabolismo , Plumas/microbiologia , Aves Domésticas , TemperaturaRESUMO
OBJECTIVES: The co-encapsulation of bioactive peptides obtained from degradation of chicken feathers and flexirubin-type pigment produced by Chryseobacterium sp. kr6 into phosphatidylcholine liposomes was investigated. RESULTS: Control empty liposomes showed mean diameter of 168.5 nm, varying to 185.4, 102.0 and 98.5 nm after the encapsulation of peptides, pigment and their co-encapsulation, respectively. Control liposomes presented zeta potential of - 20.9 mV, while the formulations containing the bioactive compounds showed values of - 30 mV or higher in magnitude. Infrared analysis revealed typical spectra for phosphatidylcholine, suggesting that no new chemical bonds were formed after encapsulation. ABTS radical scavenging assay showed that the antioxidant activity of the compounds was maintained after encapsulation. CONCLUSIONS: Feather waste can be a valuable substrate for simultaneous production of antioxidant peptides and pigment by Chryseobacterium sp. kr6, and their encapsulation into liposomes may be a suitable alternative for delivery of these natural antioxidants.
Assuntos
Antioxidantes/química , Chryseobacterium/crescimento & desenvolvimento , Plumas/microbiologia , Polienos/química , Animais , Antioxidantes/farmacologia , Biotransformação , Cápsulas , Chryseobacterium/metabolismo , Corantes/química , Composição de Medicamentos , Plumas/química , Lipossomos/química , Tamanho da Partícula , Fosfatidilcolinas/químicaRESUMO
O trabalho objetivou identificar possíveis contaminações microbiológicas no setor produtivo de avicultura de postura agroecológica do Colégio Técnico da Universidade Rural CTUR. As análises microbiológicas comprovaram a presença do fungo do gênero Cladosporium spp. os quais são altamente eficientes em produzir contaminações em ovos, sendo um potencial risco à saúde dos consumidores. Porém quando as amostras foram analisadas isoladamente de penas e ovos foi observado, mais uma vez, que havia crescimento de Cladosporium spp no isolado das penas das aves, mas não havia nenhum crescimento em nenhuma das amostras de ovos coletadas. Ou seja, não foi possível observar a infecção de forma vertical, que seria a ave transmitindo o fungo para o ovo durante a sua formação. Portanto, o setor produz ovos agroecológicos em nível seguro para o consumo.
Assuntos
Feminino , Animais , Aves Domésticas/microbiologia , Microbiologia de Alimentos , Ovos/microbiologia , Plumas/microbiologia , CladosporiumRESUMO
O trabalho objetivou identificar possíveis contaminações microbiológicas no setor produtivo de avicultura de postura agroecológica do Colégio Técnico da Universidade Rural CTUR. As análises microbiológicas comprovaram a presença do fungo do gênero Cladosporium spp. os quais são altamente eficientes em produzir contaminações em ovos, sendo um potencial risco à saúde dos consumidores. Porém quando as amostras foram analisadas isoladamente de penas e ovos foi observado, mais uma vez, que havia crescimento de Cladosporium spp no isolado das penas das aves, mas não havia nenhum crescimento em nenhuma das amostras de ovos coletadas. Ou seja, não foi possível observar a infecção de forma vertical, que seria a ave transmitindo o fungo para o ovo durante a sua formação. Portanto, o setor produz ovos agroecológicos em nível seguro para o consumo.(AU)
Assuntos
Animais , Feminino , Plumas/microbiologia , Ovos/microbiologia , Aves Domésticas/microbiologia , Microbiologia de Alimentos , CladosporiumRESUMO
Pigments synthesised by Chryseobacterium sp. kr6 growing on feather waste were extracted and characterised. The pigment extract was characterised by KOH test, UV-vis, CIELAB colour system, HPLC-DAD-MS, FTIR and its antioxidant capacity was evaluated. A positive bathochromic shift was observed when kr6 colonies or pigment extracts were subjected to alkaline solution (20% KOH) and a λmax at 450 nm was detected for acetone extracts, although no typical fine structure of carotenoids was detected in the electomagnetic spectra. The HPLC profile of the extracted pigment showed that the compound has three different peaks with λmax near 450 nm. The FTIR analysis shows some principal functional groups from a flexirubin-like molecule. The pigmented compound also presents antioxidant activity evaluated by the scavenging of the ABTS radical.
Assuntos
Antioxidantes/metabolismo , Chryseobacterium/metabolismo , Pigmentos Biológicos/química , Pigmentos Biológicos/metabolismo , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Carotenoides/análise , Carotenoides/química , Fracionamento Químico , Galinhas , Cromatografia Líquida de Alta Pressão , Chryseobacterium/química , Cor , Plumas/microbiologia , Espectrometria de Massas , Estrutura Molecular , Pigmentos Biológicos/farmacologia , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The feather is a valuable by-product with a huge annual yield produced by the poultry industry. Degradation of feathers by microorganisms is a prerequisite to utilize this insoluble protein resource. To improve the degrading efficiency of feathers, mutagenesis of the bacterium Bacillus subtilis S1-4 was performed. By combining ultraviolet irradiation and N-methyl-N'-nitro-N-nitrosoguanidine treatment for mutagenesis, a high protease-producing mutant (UMU4) of B. subtilis S1-4 was selected, which exhibited 2.5-fold higher extracellular caseinolytic activity than did the wild-type strain. UMU4 degraded chicken feathers more efficiently, particularly for the release of soluble proteins from the feathers, compared to the wild-type strain. Furthermore, an extracellular protease with a molecular weight of 45 kDa, as determined by SDS-PAGE, was purified from UMU4. Biochemical characterization indicated that the caseinolytic activity of the protease was largely inhibited by phenylmethanesulfonyl fluoride, suggesting that the purified enzyme is a serine protease. This protease was highly active over a wide range of pHs (6.0 to 12.0) and temperatures (50° to 75°C) with an optimal pH and temperature of 8.0 and 65°C, respectively. The purified enzyme exhibited good thermostability with a 72.2 min half-life of thermal denaturation at 60°C. In addition, this protease was not sensitive to heavy metal ions, surfactants, or oxidative reagents. In conclusion, strain improvement for protease production can serve as an alternative strategy to promote feather degradation. The UMU4 mutant of B. subtilis and its serine protease could be potentially used in various industries.
Assuntos
Bacillus subtilis/enzimologia , Plumas/microbiologia , Serina Proteases/metabolismo , Animais , Bacillus subtilis/genética , Bacillus subtilis/isolamento & purificação , Bacillus subtilis/efeitos da radiação , Galinhas , Plumas/metabolismo , Mutagênese , Serina Proteases/química , Serina Proteases/genética , Raios UltravioletaRESUMO
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.
Assuntos
Queratinas/metabolismo , Micrococcus luteus/enzimologia , Micrococcus luteus/metabolismo , Peptídeo Hidrolases/metabolismo , Animais , Biodegradação Ambiental , Galinhas/microbiologia , Plumas/microbiologia , Micrococcus luteus/isolamento & purificação , NADH NADPH Oxirredutases/metabolismo , Oxirredução , Aves Domésticas/microbiologia , Compostos de Enxofre/metabolismo , Gerenciamento de ResíduosRESUMO
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
Assuntos
Animais , Queratinas/metabolismo , Micrococcus luteus/enzimologia , Micrococcus luteus/metabolismo , Peptídeo Hidrolases/metabolismo , Biodegradação Ambiental , Galinhas/microbiologia , Plumas/microbiologia , Micrococcus luteus/isolamento & purificação , NADH NADPH Oxirredutases/metabolismo , Oxirredução , Aves Domésticas/microbiologia , Compostos de Enxofre/metabolismo , Gerenciamento de ResíduosRESUMO
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