Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Abstract Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

Resumo A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Degradation of keratin substrates by keratinolytic fungi

Background: The hydrolysis of keratin wastes by microorganisms is considered a biotechnological alternative for recycling and valorization through keratinolytic microorganisms. Despite their resistant structure, keratin wastes can be efficiently degraded by various microorganisms through the secretion of keratinases, which are promising enzymes for several applications, including detergents, fertilizers, and leather and textile industry. In an attempt to isolate keratinolytic microorganisms that can reach commercial exploitation as keratinase producers, the current work assesses the dynamics of keratin biodegradation by several keratinolytic fungal strains isolated from soil. The activity of fungal strains to degrade keratin substrates was evaluated by SEM, FTRIR-ATR spectra and TGA analysis. Results: SEM observations offered relevant information on interactions between microorganism and structural elements of hair strands. FTIR spectra of the bands at 1035­1075 cm-1 assigned to sulfoxide bond appeared because of S­S bond breaking, which demonstrated the initiation of keratin biodegradation. According to TGA, in the second zone of thermal denaturation, where keratin degradation occurs, the highest weight loss of 71.10% was obtained for sample incubated with Fusarium sp. 1A. Conclusions: Among the tested strains, Fusarium sp. 1A was the most active organism in the degradation process with the strongest denaturation of polypeptide chains. Because keratinolytic microorganisms and their enzymes keratinases represent a subject of scientific and economic interest because of their capability to hydrolyze keratin, Fusarium sp. 1A was selected for further studies.

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.

Dermatophytes and other associated fungi in patients attending to some hospitals in Egypt

Dermatophytes are keratinophilic fungi that infect keratinized tissues causing diseases known as dermatophytoses. Dermatophytes are classified in three genera, Epidermophyton, Microsporum, and Trichophyton. This investigation was performed to study the prevalence of dermatomycosis among 640 patients being evaluated at the dermatology clinics at Kasr elainy, El-Husein and Said Galal hospitals in Cairo and Giza between January 2005 and December 2006. The patients were checked for various diseases. Tinea capitis was the most common clinical disease followed by tinea pedis and tinea corporis. Tinea cruris and tinea unguium were the least in occurrence. Tinea versicolor also was detected. The most susceptible persons were children below 10 years followed by those aged 31–40 years. Unicellular yeast was the most common etiological agent and T. tonsuranswas the second most frequent causative agent followed by M. canis.

Fatores associados à prática do aleitamento materno exclusivo por pelo menos seis meses no estado de Pernambuco / Factors associated with breastfeeding practice for at least six months in the state of Pernambuco, Brazil

INTRODUÇÃO: Apesar do consenso científico sobre os benefícios que a amamentação proporciona à mãe, à criança, à família e ao próprio meio ambiente, além da recomendação para que sua prática seja realizada de forma exclusiva nos seis primeiros meses de vida, essa conduta está longe de ser alcançada. OBJETIVO: Analisar os fatores associados à amamentação exclusiva (AME) por pelo menos seis meses, em contraponto ao desmame total até o segundo mês de vida no estado de Pernambuco. MÉTODO: Estudo caso-controle reunindo 124 casos (AME por pelo menos seis meses) pareados por idade e sexo com 248 controles (desmame total até o segundo mês). Casos e controles foram oriundos da III Pesquisa Estadual de Saúde e Nutrição. Foram selecionadas como variáveis de exposição: idade e escolaridade materna, renda familiar, zona de moradia, consultas pré-natais, tipo de parto e profissional que o assistiu e orientação sobre amamentação no pré-natal. Foi aplicada regressão logística nas variáveis que apresentaram um valor de p < 0,2 nas análises bivariadas, adotando para a inclusão no modelo final o nível de significância p < 0,05. RESULTADOS: Dos 8 agrupamentos de variáveis consideradas como possíveis preditoras do AME por pelo menos 6 meses, mantiveram-se como fatores associados a idade materna entre 20 - 35 anos, sendo a odds ratio (OR) 2,5 e o intervalo de confiança de 95% (IC95%) 1,4 - 4,5; e a escolaridade de 5 - 8 anos de estudo (OR 2,1; IC95% 1,2 - 3,6). CONCLUSÃO: O estudo mostra que ainda são necessárias mobilizações dos poderes públicos e estímulo às pesquisas em prol do sucesso do AME e da saúde materno-infantil. .

INTRODUCTION: Despite the scientific consensus on the benefits that breastfeeding provides for the mother, the baby, the family and the environment, and also the recommendation to breastfeed exclusively for six months, this practice is far from being achieved. OBJECTIVE: To analyze the factors associated with exclusive breastfeeding (EBF) for at least six month, as opposed to weaning up to the second month of life in the state of Pernambuco, Brazil. METHODS: A case-control study of 124 cases (EBF for at least six months) matched for age and sex with 248 controls (weaning up to the second month of life). Cases and controls were drawn from the III State Health and Nutrition Survey. The exposure variables selected were maternal age and education, per capita income, housing zone, prenatal consultations, type of delivery, professional who assisted the delivery, and prenatal breastfeeding guidance. Logistic regression was applied to variables that showed a p-value < 0.2 in the bivariate analysis, and the variables with p-value < 0.05 were included in the final model. RESULTS: Of the eight groups of variables considered as possible predictors of EBF for at least six months, two remained as associated factors: maternal age between 20 - 35 years old, with odds ratio (OR) 2.5 and 95% confidence interval 95%CI 1.4 - 4.5; and maternal education of 5 - 8 years of schooling (OR 2.1; 95%CI 1.2 - 3.6). CONCLUSION: The study shows that mobilization of the public sector and stimulus to research is still needed for the success of EBF and for mother and child health. .

Cytokeratin expression in corneal dystrophies / Expressão de citoqueratinas em distrofias corneanas

PURPOSE: To identify an immunohistochemical pattern of epithelial markers in granular, lattice and Avellino corneal dystrophies. METHODS: Twenty-two corneal buttons, diagnosed as lattice (17), Avellino (4) and granular (1) underwent immunohistochemical studies of cytokeratins (CKs) on pa- raffin-embedded sections (group I). Monoclonal antibodies for pan-CK (AE1/AE3) and CKs 3/12, 5/6, 8, 18 and 19 were used. Twenty-two normal corneas were used as the control (group II). RESULTS: Six lattice and 2 Avellino cases of group I stained positively with anti-CK 3/12 in corneal epithelium and areas of corneal stroma deposits. One of these cases of lattice was positive for anti-pan-CK (AE1/AE3) also in the epithelium and areas of corneal stroma deposits with a similar pattern. None of the controls (group II) revealed any staining in corneal stroma. All disease and control cases (groups I and II) revealed positive staining in corneal epithelium. CONCLUSION: AE1/AE3 and CK 3/12 anti-CK positive markers in the stromal deposits of lattice and Avellino dystrophies may suggest an epithelial genesis of the disease.

OBJETIVO: Investigar a expressão de citoqueratinas (CKs) em córneas com distrofias corneanas tipo granular, lattice e Avellino. MÉTODOS: Vinte e dois botões corneanos com diagnóstico anatomopatológico de distrofia estromal tipo lattice (17), Avellino (4) e granular (1) foram submetidos à avaliação imunohistoquímica nos tecidos inclusos em parafina (grupo I). Anticorpos monoclonais para pan-CK (AE1/AE3) e CKs de números 3/12, 5/6, 8, 18 e 19 foram utilizados. Vinte e dois botões corneanos normais foram usados como controle (grupo II). RESULTADOS: Oito casos do grupo I (seis lattice e dois Avellino) apresentaram reações imuno-histoquímicas positivas com anti-CK 3/12, tanto no epitélio como nos depósitos estromais e um destes casos (lattice) também se mostrou positivo para anti-pan-CK (AE1/AE3) com o mesmo padrão de reação. Nenhum caso do grupo II mostrou reação imuno-histoquímica positiva no estroma corneano. Na avaliação imuno-histoquímica dos grupos I e II, o epitélio apresentou uma reação positiva com o anticorpo anti-pan-CK (AE1/AE3) e com o anti-CK 3/12. CONCLUSÃO: O fato da pan-CK e CK 3/12 apresentarem uma reação positiva nos depósitos das distrofias tipo lattice e Avellino sugere uma origem epitelial desses depósitos corneanos.

Análise imuno-histoquímica das citoqueratinas em ameloblastoma e tumor odontogênico adenomatóide / Immunohistochemical analysis of cytokeratins in ameloblastoma and adenomatoid odontogenic tumor

OBJETIVO: O presente trabalho teve por objetivo traçar o perfil das citoqueratinas (CKs) 7, 8, 10, 13, 14, 18 e 19 em ameloblastomas e tumor odontogênico adenomatóide (TOA) visando contribuir para o entendimento da histogênese desses tumores e somar com os resultados já relatados na literatura. MATERIAL E MÉTODO: do arquivo do Laboratório de Anatomia Patológica do Departamento de Odontologia da Universidade Federal do Rio Grande do Norte (UFRN) foi selecionada uma amostra com dez casos de ameloblastomas e oito de TOA para o estudo imuno-histoquímico, utilizando-se anticorpos anti-CKs pelo método da estreptoavidina-biotina. RESULTADOS: Observou-se que nos ameloblastomas a CK 14 esteve presente em todos os casos, enquanto a CK 19 foi observada nas células periféricas (oito casos) e nas centrais (cinco casos). Para os TOA, observou-se imunopositividade para a CK 14 em todos os casos, enquanto a CK 19 esteve marcada predominantemente nas células ductais (seis casos). CONCLUSÃO: As citoqueratinas são expressas de forma variada nos ameloblastomas e nos TOA, os quais preservam CK típicas do germe dental em estágios avançados do desenvolvimento, confirmando sua origem exclusiva a partir do epitélio odontogênico e não se evidenciando CK características do epitélio escamoso.

OBJECTIVES: The aim of the present study was to describe the immunohistochemical expression of cytokeratins (CKs) 7, 8, 10,13, 14, 18 and 19 in the epithelial components of ameloblastomas and adenomatoid odontogenic tumor (AOT). The results were compared and histogenesis discussed. MATERIAL AND METHOD: Specimens of ten ameloblastomas and eight adenomatoid odontogenic tumors were examined by immunohistochemistry using streptavidin-biotin-peroxidase complex method and anti-CKs antibody. The sample was obtained from Department of Oral Pathology, Federal University of Rio Grande do Norte. RESULTS: Immunohistochemical reactivity for CK14 was detected in all cases of ameloblastomas, and the expression of CK19 was diffusely present in peripheral cells (eight cases) and in central cells of five ameloblastomas. In AOTs, immunopositivity for CK14 was detected in all cases, whereas CK 19 was predominantly expressed in ductal cells (six cases). CONCLUSION: The CKs was expressed by several forms in ameloblastomas and AOTs, that have shown typical CKs of dental germ suggesting that these tumors have odontogenic epitheilial differentiation, whereas it has not evinced proper CKs of squamous epithelial.

Carcinoma de células de Merkel de labio: presentación de un caso con evolución especial y diagnóstico diferencial inmunohistoquímico / Lip Merkel cell carcinoma: presentation of a case with special evolution and immunohistochemic differential diagnosis

El carcinoma de células de Merkel es un tumor neuroendócrino infrecuente y muy agresivo a la piel. Las lesiones se presentan en zonas expuestas al sol tal como cabeza y cuello, extremidades y tronco; se han comunicado localizaciones intraorales y en labios. El diagnóstico diferencial clínico y por histología convencial, con otros tumores, es muy dificultoso. La inmunohistoquímica resulta útil para un correcto diagnóstico dado el inmunofenotipo específico de este tumor. El conocimiento por parte del clínico y del cirujano de la clínica y del inmunofenotipo diagnóstico por parte del patólogo, es esencial para un diagnóstico de certeza, y la clave para el planeamiento del tratamiento adecuado.

Presencia de condensaciones pericanaliculares inmunogénicamente semejantes a cuerpos de Mallory en colestasia / Presence of perinacular condensations immunogenicaly similar to Mallory's bodies in cholestasia

El rol del citoesqueleto del hepatocito en secreción biliar y en la génesis de colestasia ha sido estudiado especialmente en relación a microfilamentos y microtúbulos, restándosele importancia los filamentos intermedios (FI). En cambio, estudios recientes han demostrado que la integridad de los FI del hepatocito es fundamental en la secreción biliar. Los FI del hepatocito corresponden a citoqueratinas y en ciertas patologías, especialmente de etiología alcohólica, éstos se alteran y en algunas ocasiones forman agregados como los cuerpos de Mallory (CM) cambiando sus características antigénicas. Se estudiaron 131 biopsias hepáticas con técnicas inmunohistoquímicas, utilizando sueros anti-queratinas epidérmicas y anti-ubiquitina, polipéptido de función proteolítica de proteínas anormales. De las biopsias estudiadas, 47 por ciento presentaban signos de colestasia y de éstas, un 64 por ciento presentó inmunorreacción en condensaciones pericanulares con anti-ubiquitina y un porcentaje algo menor con anti-queratinas; el resto de las biopsias fueron negativas con ambos anticuerpos. Estas observaciones indican que en biopsias hepáticas con signos de colestasia, las condensaciones pericaniculares correspondían a FI alterados, con características inmunogénicas semejantes a CM