Thesis (D. Min.)--Westminster Theological Seminary, Philadelphia, 2003. / Includes vita. Includes bibliographical references (leaves 261-267).

Made available in DSpace on 2016-12-08T16:24:18Z (GMT). No. of bitstreams: 1 PGCA14MA139.pdf: 23015 bytes, checksum: 79359edb3078879f5ca9aa0338da2c45 (MD5) Previous issue date: 2013-03-01 / With the objective of evaluate the effect of continuous and intermittent lighting programs on the production and quality egg of laying hens in the early lay pullets rearing in open poultry house, a trial was conducted at Aviculture Section of Santa Catarina State University, Lages, SC, Brazil. Were used 420 laying hens with 21 weeks during seven periods of 28 days, they were submitted to three different lighting programs in a completely randomized design with 10 replications. The continuous program with 16 hour photoperiod (daylight + artificial light) and escotoperiod eight hours. The intermitente lighting program of 30 minutes had two photophases of 30 min between natural light and main night eight hours and two escotofases and intermitente lighting of 15 minutes, this program is similar to the previous program, with two photophases 15 min between natural light, a main night eight hours and two escotofases. The avaliation was based on the feed intake (g/bird/day), egg production (%/bird/day), egg weight (g), feed conversion (g/g), egg mass (g), specific gravity (g/cm³), albumen height (mm), Haugh unit, egg componente yield (%), organ index (%) and the means compared by Tukey test (5%). The results presented for feed conversion, specific gravity, weight of fresh egg and stored for 14 and 28 days, egg yield componentes and organs index showed no difference (P>0.05). The intermitente lighting programs 30 minutes and 15 minutes showed significant differences (P<0.05) for feed intake in the second, sixth and seventh trial period, but no significant differences (P>0.05) in egg production. The egg weight, egg mass, showed no significant difference (P>0.05) as well as albumen height and Haugh unit showed no significant difference (P>0.05) for continuous and intermitente lighting programs 30 min and 15 min. Intermittent lighting programs applied in laying hens in the early lay pullets rearing in open poultry house not adversely affect the production performance, egg quality and internal organs of birds, as well as shorten the use of artificial lighting, reducing energy consumption / Com o objetivo de avaliar o efeito de programas de iluminação contínuo e intermitentes sobre produção e qualidade de ovos de poedeiras comerciais semipesadas em início de postura criadas em galpão aberto, realizou-se um trabalho no Setor de Avicultura da Universidade do Estado de Santa Catarina, Lages - SC. Utilizou-se 420 galinhas de 21 semanas de idade durante sete períodos de 28 dias, as quais foram submetidas a três diferentes programas de iluminação em um delineamento inteiramente ao acaso com 10 repetições. O programa contínuo contou com fotoperíodo de 16 horas (luz natural + luz artificial) e escotoperíodo de oito horas. O programa de iluminação intermitente 30 minutos teve duas fotofases de30min entre a luz natural e uma noite principal de oito horas e duas escotofases e o programa de iluminação intermitente 15 minutos, semelhante ao programa anterior, com duas fotofases de 15min entre a luz natural, uma noite principal de oito horas e duas escotofases. Avaliou-se o consumo de ração (g/ave/dia), produção de ovos (%/ave/dia), peso dos ovos (g), conversão alimentar (g/g), massa de ovos (g), gravidade específica (g/cm³), altura de albúmen (mm), unidade Haugh, rendimento dos componentes do ovo (%), índice de órgãos (%) sendo as médias comparadas pelo teste Tukey (5%). Os resultados apresentados para conversão alimentar, gravidade específica, peso de ovo fresco e armazenado por 14 e 28 dias, rendimento dos componentes do ovo e índices de órgãos não apresentaram diferença (P>0,05). Os programas de iluminação intermitente 30 min e intermitente 15 min apresentaram diferenças significativas (P<0,05) para consumo de ração no segundo, sexto e sétimo período experimental, porém não apresentaram diferenças significativas (P>0,05) na produção de ovos. O peso de ovo, massa do ovo, não apresentou diferença significativa (P>0,05), assim como altura de albúmen e unidade Haugh não apresentaram diferença significativa (P>0,05) para os programas de iluminação contínuo e intermitentes 30min e 15min. Programas de iluminação intermitentes aplicados em poedeiras semipesadas ao início da postura criadas em galpões abertos não afetam negativamente o desempenho zootécnico, a qualidade de ovos e órgãos internos das aves, além de diminuir o tempo de uso de iluminação artificial, reduzindo o consumo de energia elétrica

I den här undersökningen tittar jag närmare på hur ordet hen används i interaktioner på Twitter. Jag frågar mig i vilka sammanhang och om vem som hen kan användas i dessa konversationer. Vad tycks gynna eller begränsa användningen av hen? Här är jag, först och främst, intresserad av att se på hur olika typer av normer kring kön och sexualitet kan tänkas påverka användningen, och vilka sådana normer som är relevanta i konversationerna. För att göra detta använder jag mig av metoden Membership categorization analysis. Med hjälp av denna metod tittar jag närmare på vilka kategoriseringar som görs relevanta och kopplas till användningen av hen. Detta möjliggör att just komma åt vilka normer som blir aktualiserade, hur de konstrueras och potentiellt påverkar användningen av hen. Därtill vill jag också se vilka möjliga alternativa positioner som hen kan tänkas etablera i relation till normativaköns- och sexualitetsdiskurser. För att fördjupa analysen av normer, kön och sexualitet tar jag min teoretiska utgångspunkt från ett queerteoretiskt perspektiv. Särskilt använder jag mig av performativitet, och det normativa ramverket den heterosexuella matrisen, som viktiga begrepp i uppsatsen för att förståhur olika köns- och sexualitetsnormer skapas och återskapas i interaktion, samt hur dessa normer påverkar möjligheterna att använda hen i konversationerna. I analysen presenterar jag två huvudgrupper som representerar olika övergripande användningsområden för hen som jag identifierat utifrån mitt material. Den ena gruppen består av användningar där hen används för att inte göra kön relevant i konversationen, och den andra är användningar där hen kan användas för att utmana eller ifrågasätta normativa föreställningar om, och beskrivningar av, kön och sexualitet. I den första gruppen är också den normkritiska aspekten av hen mer begränsad, medan den i den andra gruppen är mer central och utmanarflera olika typer av könsrelaterade normer. Ett övergripande resultat är således att hen, beroende på samtalskontexten både kan begränsasav olika könsnormer, och innebära möjligheten att dessa normer utmanas. En ytterligare aspekt som tycks begränsa möjligheten att använda hen är huruvida den som omnämns med detta ord har en relationell närhet till personen som använder ordet. I de mer nära relationerna tenderar könande pronomen att användas och tvärtom kan i stället hen användas för att benämna de personer som inte framträder med samma närhet till personen som gör referensen.

Den här uppsatsen undersöker hur utbrett bruket av en som könsneutralt generaliserande pronomen är bland anställda vid järnvägen i Mellansverige och om det skiljer sig åt beroende på utbildningsbakgrund, ålder eller kön. Uppsatsen avser även att undersöka om de som redan använder hen som könsneutralt personligt pronomen och/eller som ett tredje kön är mer benägna att också använda en. Dessutom undersöks om bruket av man respektive en skapar konceptualiseringar av olika kön. Undersökningen genomförs med en kvantitativ enkätundersökning med kvalitativa inslag och genom kvalitativa intervjuer. Resultaten visar att bruket inte är utbrett men att majoriteten har en positiv attityd till att en kan användas som könsneutralt generaliserande pronomen. De få som faktiskt använder en representeras framförallt av yrkesutbildade, personer födda 1985 och senare samt av kvinnor där samtliga som medvetet använder en även använder hen. Man liksom en skapar inte konceptualiseringar av något specifikt kön för en klar majoritet i enkätundersökningen, i de få fall det görs skapar man konceptualiseringar av könen kvinna och man medan en skapar konceptualiseringar av hen. Intervjuerna visar på ett annorlunda resultat där texten med man skapar konceptualiseringar av inget specifikt kön, båda könen och man medan en skapar konceptualiseringar av flera kön, båda könen och kvinna. Slutsatser är att en uppfattas mest dialektalt, minst politiskt, att attityden till en är god även om bruket inte är utbrett och att hen manar till ytterligare könsneutrala bruk. Varken man eller en skapar konceptualiseringar av kön för de flesta, man tenderar dock att reproducera ett binärt könssystem medan en inkluderar fler könsidentiteter.

In Deutschland werden jährlich über 40 Millionen männliche Eintagsküken aus Legelinien aufgrund vorrangig wirtschaftlicher Interessen getötet. Dies stellt sowohl ein ethisches als auch ein tierschutzrechtliches Problem dar (ANON. 2006, IDEL 2007). Gerade vor dem Hintergrund aktueller politischer Entscheidungen (MUNLV NRW 2013, NI MELV 2014) besteht ein Bedarf an Alternativen zur Tötung männlicher Eintagsküken. Verschiedene Lösungsansätze wie z. B. das Zweinutzungshuhn (ICKEN et al. 2013) oder aber die Mast männlicher Geschwisterhühner (KAUFMANN und ANDERSSON 2013) sind derzeit aus ökonomischen und ökologischen Gründen nicht flächendeckend realisierbar. Eine weitere Möglichkeit bietet die In-ovo-Geschlechtsbestimmung. Hierbei wird das embryonale Geschlecht bereits vor dem Schlupf identifiziert; nachfolgend können die Eier mit männlichen Embryonen aussortiert werden. Um sowohl ethischen als auch tierschutzrechtlichen Aspekten Genüge zu tun, sollte die Geschlechtsidentifikation dabei vor Einsetzen des embryonalen Schmerzempfindens stattfinden (Tag 10 + 12 h der Bebrütung; CLOSE et al. 1997). Ziel der vorliegenden Arbeit war die Entwicklung einer verlässlichen Methode zur In-ovo-Geschlechtsbestimmung anhand geschlechtsspezifischer Differenzen im Hormongehalt der Allantoisflüssigkeit sieben bis zehn Tage alter Hühnerembryonen. Nachfolgend wurde der Einfluss der Geschlechtsbestimmung auf die embryonale Entwicklung, Schlupferfolg, Aufzucht sowie die Leistungsparameter der adulten Tiere analysiert. Im Rahmen der ersten Teilstudie erfolgte die Beprobung von n = 750 Eiern des Braunlegehybrids Lohmann Brown (LB, Lohmann Tierzucht GmbH, Deutschland). Der minimalinvasiven Entnahme von Allantoisflüssigkeit folgte die Untersuchung auf 17β-Östradiol (E2), Östronsulfat (E1S) und Testosteron mittels an das Haushuhn angepassten Enzymimmunoassays (ELISA). Es konnten sowohl für E2 als auch für E1S signifikante (p < 0,01) geschlechtsspezifische Differenzen in der Allantoisflüssigeit von neun und zehn Tage alten Embryonen nachgewiesen werden. Die Testosteronkonzentration hingegen zeigte an keinem der untersuchten Tage geschlechtsabhängige Unterschiede und erwies sich somit für die In-ovo-Geschlechtsbestimmung als ungeeignet. Die statistische Auswertung ergab, dass die Bestimmung von E1S eine frühere und genauere Geschlechtsidentifikation ermöglicht als die von E2. Der für E1S festgelegte Grenzwert erreicht bei neun Tage alten Embryonen eine 86%ige Sensitivität und 83%ige Spezifität. In der zweiten Teilstudie wurde die zuvor etablierte Technik der Geschlechtsbestimmung mittels E1S an 8 + 4 h (n = 2420) und 9 + 4 h (n = 2850) Tage alten Embryonen der Herkunft LB sowie an n = 150 9 + 4 h alten Embryonen des Weißlegehybrids Lohmann Selected Leghorn (LSL, Lohmann Tierzucht GmbH, Deutschland) überprüft. Das Geschlecht der 8 + 4 h Tage alten Embryonen konnte zu 84 % korrekt identifiziert werden. Dieser Wert stieg bei 9 + 4 h Tage alten Embryonen auf 98 % (LB) bzw. 100 % (LSL) an. Im Vergleich zu einer unbehandelten Kontrollgruppe (n = 5258) wurde die Schlupfrate durch die Entnahme von Allantoisflüssigkeit um 1,4 - 3,5 (LB) bzw. 12,7 Prozentpunkte (LSL) reduziert. Nachfolgend wurden 150 Tiere der Versuchsgruppe und 80 Tiere der Kontrollgruppe für eine Aufzuchtperiode von 17 Wochen eingestallt. Hierbei zeigten sich hinsichtlich des Körpergewichtes signifikante (p < 0,05) Unterschiede zwischen Versuchs- und Kontrollgruppe in Woche 4 und 6, wobei die Zunahmen in der Versuchsgruppe geringer waren. Anschließend wurde die Leistung von 120 Tieren der Versuchsgruppe und 60 Tieren der Kontrollgruppe bis Lebenswoche 33 bezüglich Legeleistung, Eigewicht, Körpergewicht sowie Futterverbrauch analysiert. Bei keinem der untersuchten Parameter konnten signifikante Unterschiede zwischen den Gruppen festgestellt werden (p > 0,05). Die Resultate der vorliegenden Arbeit zeigen, dass eine verlässliche Geschlechtsbestimmung in ovo bei 9 + 4 h Tage alten Hühnerembryonen mithilfe einer Bestimmung der E1S-Konzentration in der Allantoisflüssigkeit möglich ist; zudem ist die beschriebene Methode bei verschiedenen Legelinien anwendbar. Die Entnahme von Allantoisflüssigkeit führt zwar zu einer minimalen Reduktion der Schlupfrate, bei adulten Legehennen kommt es jedoch zu keiner Beeinträchtigung der Produktionsleistung. Demnach erfüllt das etablierte Verfahren alle Grundvoraussetzungen für eine Anwendung in kommerziellen Brütereien. Da die Geschlechtsbestimmung vor Einsetzen des embryonalen Schmerzempfindens erfolgt, kann sie somit als Grundlage für eine ethisch vertretbare Alternative zum Töten männlicher Eintagsküken angesehen werden. / In Germany about 40 million day-old male chicks are culled each year predominantly because of economic reasons. From the animal welfare as well as the ethical point of view this is a problematic situation (ANON. 2006, IDEL 2007). Particularly with regard to current political decisions (MUNVL NRW, NI MELV 2014) alternatives to the culling of male day-old chicks are required. Different approaches such as a dual-purpose breed (ICKEN et al. 2013) or the fattening of male layer-hybrids (KAUFMANN and ANDERSSON 2013) are not ubiquitous marketable at present due to economic and ecological reasons. In ovo sexing represents another option; the embryonic gender is determined before hatch and the eggs containing male embryos can be eliminated subsequently. To comply with ethical and animal welfare aspects, the sexing should take place before the onset of embryonic pain perception (embryonic day 10 + 12 h; CLOSE et al. 1997). Aim of this thesis was the development of a reliable method for in ovo gender identification with the help of sex-specific differences in the hormone concentration of the allantoic fluid of seven to ten day old chick embryos. Subsequently, the influence of gender identification on embryonic development, hatching rate, rearing as well as production performance of the adult hens was analysed. Within the first study n = 750 eggs of the brown layer-hybrid Lohmann Brown (LB; Lohmann Tierzucht GmbH, Germany) were sampled for allantoic fluid. After the minimally invasive withdrawal the allantoic fluid was analysed via enzyme immunoassays (ELISA) adapted to domestic chicken for 17β-oestradiol (E2), oestrone sulphate (E1S) and testosterone. With regard to E2 and E1S, significant (P < 0.01) sex-specific differences were observed in the allantoic fluid of nine and ten day old embryos. Testosterone on the other hand displayed no gender-related variances on any of the analysed days. Therefore, it proved to be unsuitable for gender identification using the method applied in this study. Statistical analysis showed that the analysis of E1S allows an earlier and more accurate sexing than the E2-assay. The limit value determined for E1S has a sensitivity of 86 % and a specificity of 83 % for nine day old embryos. The previously established method for gender identification via E1S detection in the allantoic fluid was verified with a larger number of samples in the second study. The allantoic fluid of day 8 + 4 h (n = 2420) and day 9 + 4 h (n = 2850) old LB embryos as well as n = 150 day 9 + 4 h old embryos of the white layer-hybrid Lohmann Selected Leghorn (LSL; Lohmann Tierzucht GmbH, Germany) was analysed. For day 8 + 4 h old embryos the sex was correctly identified in 84 %. The accuracy of gender prediction increased for day 9 + 4 h old embryos up to 98 % (LB) and 100 % (LSL). Compared to an untreated control group (n = 5258) sampling of allantoic fluid reduced the hatching rate by 1.4 - 3.5 (LB) and 12.7 points of percentage (LSL). In the following, 150 animals of the experimental group and 80 animals of the control group were reared for a period of 17 weeks. With regard to the body weight significant differences (P < 0.05) were observed in weeks 4 and 6, with the animals of the experimental group having a lower body weight. Subsequently the production performance of 120 hens from the experimental and 60 hens from the control group was analysed up to an age of 33 weeks. With respect to egg production, egg weight, body weight and feed consumption no significant differences (P > 0.05) were observed between the groups. The results of this thesis demonstrate that a reliable in ovo sexing of day 9 + 4 h old chicken embryos is possible via the measurement of E1S in the allantoic fluid; additionally the method is not limited to a certain layer strain. The sampling of allantoic fluid reduces the hatching rate only marginally. The production performance of adult hens on the other hand is not affected. Therefore, the described technique fulfils all the basic requirements for an alternative method to the culling of day-old male layer chicks. Because gender identification takes place before the onset of embryonic pain perception it can serve as the basis for an ethical alternative to the culling of male day-old chicks from layer-hybrids.

My research was focused on greater sage-grouse (Centrocercus urophasianus; hereafter "sage-grouse") nest-site selection, nest success, and hen survival in relation to avian predators. The trade-off between using habitat and avoiding predators is a common decision for prey species including sage-grouse. In Chapter 2, I compared avian predator densities at sage-grouse nest and brood locations to random locations. Sage-grouse were located where densities of small, medium, and large avian predators were 65-68% less than random locations. The effects of anthropogenic and landscape features on habitat use of sage-grouse hens have not been evaluated relative to avian predator densities. In Chapter 3, I compared anthropogenic and landscape features and avian predator densities among sage-grouse locations (nest, early-brood, late-brood) and random locations. I found sage-grouse hens chose locations with lower avian predator densities compared to random locations, and selected locations farther away from anthropogenic and landscape features. Depredation of sage-grouse nests can be an influential factor limiting their productivity. Predator removal has been simultaneously proposed and criticized as a potential mitigation measure for low reproductive rates of sage-grouse. In Chapter 4, I hypothesized that sage-grouse nest success would be greater in areas where Wildlife Services lowered common raven (Corvus corax: hereafter "raven") density. I found that Wildlife Services decreased raven density by 61% during 2008-2011 but I did not detect a direct improvement to sage-grouse nest success. However, sage-grouse nest success was 22% when ravens were detected within 550 m of a sage-grouse nest and 41% when no raven was detected within 550 m. In Chapter 5, I assessed interactive effects of corvid densities relative to anthropogenic and landscape features on sage-grouse nest success. I found that sage-grouse nest success was positively correlated with rugged habitat. Survival of breeding-age birds is the most important demographic parameter driving sage-grouse abundance. In Chapter 6, I evaluated the effect of raptor densities, proximity to anthropogenic and landscape features, and hen behavior on survival of sage-grouse hens. I found that sage-grouse hen survival was negatively correlated with golden eagle (Aquila chrysaetos) density, proximity to anthropogenic and landscape features, and hen parental investment (nesting and brood-rearing).

In layer hen commercial production systems, chicks are not raised by hens and do therefore not receive any maternal care. In the wild, after hatching, chicks undergo filial imprinting where they create a social bond with a stimulus in their surrounding environment, usually the mother hen. Filial imprinting behaviors remain in chicks until adulthood. This bond and maternal care influence the emotional development of chicks. Here, I aimed to investigate short- and long-term effects of filial imprinting on a model hen, on the emotional state of White Leghorn chicks, hatched at Linköping University. To measure optimism, pessimism, and fear, a judgment bias test (JBT), an open field test (OFT), and a novel object test (NOT), was performed and replicated. Before the tests, half of the chicks were imprinted on a model hen, while the other half was not given such a model to imprint on. All chicks were weighed during the entire project. The hypotheses were that the imprinted chicks would be more optimistic in the JBT, show less fearful behavior in the OFT and NOT, and weigh more compared to the non-imprinted chicks. If these results would be found, imprinting could potentially be used to reduce stress in commercially hatched chicks, and thus improve their welfare. The results were inconsistent with all three hypotheses. This indicates that imprinting on a model hen, did not have a positive impact on the chicks’ emotional state. Further studies need to be performed to find methods to improve commercially hatched chicks’ emotional welfare.

Les sites industriels abandonnés présentent souvent des sols ayant des propriétés physico-chimiques atypiques et des concentrations élevées en métaux traces, rendant leur dépollution coûteuse de surcroît quand les superficies sont grandes. Les phytotechnologies, comme la phytoremédiation, émergent alors comme des options économiques et socialement acceptables. Cette étude, conduite en conditions semi-contrôlées, a examiné la capacité de saules (Salix miyabeana 'SX67') et de peupliers (Populus DNxM '915508') à phytoremédier des sols contaminés, notamment par le cuivre, et a comparé leurs performances suivant l’application de frass, un résidu de l’élevage d’insectes, et de fumier de poule, tous deux produits dans une démarche d’économie circulaire. Dans l'ensemble, aucune tendance claire n'a pu être identifiée entre les traitements et les espèces concernant la production de biomasse, les paramètres physiologiques, ou l'assimilation des éléments traces. Cependant, les saules amendés avec du frass ont atteint une hauteur moyenne de 210 cm, significativement supérieure à celle des saules non fertilisés. Les concentrations de cuivre extrait par les plantes n'ont pas été affectées par les deux types d'amendements organiques. Comme pour le cuivre, qui a montré des concentrations de 407 mg kg-1 dans les racines et de 14 mg kg-1 dans les parties aériennes, la majorité des dix éléments étudiés a été principalement retrouvée dans les racines. Seuls le zinc et le cadmium ont montré un facteur de translocation supérieur à un. En ce qui concerne les différentes fractions de cuivre dans le sol, aucune différence significative n'a été observée entre les traitements pour les fractions labiles et phytodisponibles, bien qu'une tendance puisse être notée pour les peupliers amendés avec du fumier. Les macronutriments tels que l'azote et le phosphore ont montré une interaction complexe impliquant les espèces, les amendements, et la contamination en cuivre. Malgré des résultats ambigus et une complexité inattendue, cette étude semble indiquer que le frass et le fumier de poule sont des matériaux efficaces en comparaison des engrais inorganiques communément utilisés. Néanmoins, ils soulignent également la nécessité d'une recherche plus approfondie sur l'utilisation de ces amendements organiques issus de l'économie circulaire afin d'optimiser leur utilisation. Une telle exploration, en alignement avec les principes de durabilité et de recyclage, pourrait éventuellement orienter nos stratégies vers des solutions plus efficaces, viables et économiques, contribuant ainsi à l'avenir de la restauration écologique des terrains industriels abandonnés. / Brownfield sites, characterized by soils with atypical physicochemical properties and high trace element (TE) concentrations, are generally expensive to decontaminate especially when large areas are involved. Economical and socially approved phytotechnologies, such as phytoremediation, offer environmentally conscious solutions that require an approach distinct from those using chemical fertilizers in industrial or agricultural contexts. This study, conducted under semi-controlled conditions, examined the ability of willows (Salix miyabeana 'SX67') and poplars (Populus DNxM '915508') to phytoremediate contaminated soils, especially those contaminated with copper, and compared their performance following the application of frass, a residue from insect breeding, and chicken manure, both produced in a circular economy approach. The study examined biomass production, specific physiological properties, and the assimilation of copper and other trace elements into plant tissues. Results showed no discernible pattern between the treatments and the species. However, willows treated with frass achieved significantly greater heights, averaging 210 cm. Neither organic amendment influenced the level of copper phytoextracted with, in most cases, higher concentrations of trace elements in roots rather than in the aerial parts, exemplified by copper levels of 407 mg kg-1 in roots and only 14 mg kg-1 in leaves. Notably, only zinc and cadmium had translocation factors greater than one. Concerning soil copper fractions, there were no statistical differences in both labile and phytoavailable copper across treatments despite a visible trend for frass-amended poplars. Macronutrients such as nitrogen and phosphorus demonstrated complex interactions dependent on the species, amendments, and Cu contamination. Despite ambiguous results and unexpected complexity, this study seems to indicate that frass and chicken manure are effective materials compared to commonly used inorganic fertilizers. However, they also emphasize the need for more in-depth research on the use of these organic amendments from the circular economy to optimize their utilization. Such exploration, in alignment with principles of sustainability and recycling, could potentially guide our strategies towards more effective, viable, and economical solutions, thereby contributing to the future of ecological restoration of abandoned industrial sites.

O aumento na procura por produtos naturais pelo mercado farmacêutico, cosmético e alimentício demanda pesquisas no desenvolvimento desses produtos. Esses são direcionados à obtenção de substâncias de origem vegetal ou animal, assim como, para produtos biotecnológicos. Investigações quanto à atividade antibacteriana de proteínas e peptídeos são realizadas. Dentre essas substancias, podemos citar as lisozimas, proteínas que hidrolisam as ligações &#946; 1-4 glicosídicas entre o ácido N-acetilmurâmico e N-acetilglicosamina, presentes no peptidoglicano da parede celular bacteriana. Além disso, apresentam atividade de quitinase, ou seja, quebram a ligação glicosídica da quitina presente na parede fúngica. As lisozimas apresentam alta especificidade pela parede microbiana indicando aparente ausência de efeitos tóxicos aos humanos. Assim, tornando-a candidata a agente antimicrobiano em formulações cosméticas e farmacêuticas. A lisozima de ovo de galinha tem atividade antimicrobiana, entretanto não havia estudos relacionados com os micro-organismos contaminantes normalmente encontrados em produtos farmacêuticos e cosméticos. Além disso, a lisozima recombinante de Musca domestica (MdL1) não possui ainda sua atividade antimicrobiana definida. Os objetivos do trabalho foram:1) Obtenção da lisozima recombinante de Musca domestica (MdL1); 2) Avaliação a atividade antimicrobiana da MdL1 e de lisozima de ovo de galinha, Hen Egg White Lysozyme (HEWL), frente à Staphylococcus aureus (ATCC 6538), Micrococcus luteus (ATCC 4698), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 8739), Candida albicans (ATCC 10231) e Aspergillus niger (ATCC 16404); 3) Avaliação da toxicidade da lisozima em cultura de células de fibroblastos (ATCC CCL-92). A MdL1 foi obtida por meio de expressão gênica em Pichia pastoris GS115 (Invitrogen), concentrada utilizando polietilenoglicol 6000 e dialisada contra água deionizada através da membrana com porosidade seletiva de 12kDa. A homogeneidade foi analisada por eletroforese em gel de poliacrilamida em condições desnaturantes; e a atividade catalítica foi avaliada utilizando células liofilizadas de Micrococcus luteus como substrato. A atividade antimicrobiana foi determinada por método específico para cada enzima. A toxicidade in vitro das amostras foi avaliada pela viabilidade celular de fibroblastos ATCC CCL-92. A MdL1 obtida apresentou características de homogeneidade adequadas e atividade de 108,35 U/mg. A HEWL mostrou-se ativa contra S. aureus, M. luteus e C. albicans. A MdL1 apresentou-se ativa contra M. luteus, apenas. Devido a ausência de atividade antimicrobiana a MdL1 não foi submetida a avaliação citotóxica. Em relação à HEWL não demonstrou citotoxicidade na avaliação prévia realizada. / The increase in the search for natural products by pharmaceutical, cosmetic and food markets requires researches in these products development. These are directed to obtaining substances of vegetal or animal origin, as well as biotechnological products. The research in relation to antibacterial activity of proteins and peptides is carried out. Among these substances, it is possible to mention the lysozyme, protein that catalyze the break of 1,4-beta-D glucosidic bond between N-acetylmuramic acid and N-acetilglicosamine which are present in peptidoglicane of the bacterial cell wall. Besides, there is kitinase activity, that is, they break the glicosidic bond of chitin which is present in fungal wall. The lysozymes show high specificity by microbial wall indicating apparent absence of toxicological effects to human beings. Therefore, it becomes the candidate to antimicrobial ingredient in cosmetic and pharmaceutical dosage forms. The hen egg white lysozyme has antimicrobial activity, however there were no studies related to spoiled microorganism usually found in pharmaceutical and cosmetic products. In addition, there were not studies about microbial activity of recombinant Musca domestica lysozyme 1 (MdL1). The aim of this research was: 1) To obtain MdL1; 2) Evaluation of MdL1 and hen egg white lysozyme (HEWL) antimicrobial activity against Staphylococcus aureus (ATCC 6538), Micrococcus luteus (ATCC 4698), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 8739), Candida albicans (ATCC 10231) e Aspergillus niger (ATCC 16404); 3) Evaluation of lysozyme toxicity in fibroblast cells culture (ATCC CCL-92). The MdL1 was expressed as recombinant protein in Pichia pastoris GS115 (Invitrogen), concentrated using polyethylene glycol 6000 and dialyzed against deionized water through the selective porosity of 12kDa membrane. The homogeneity was analyzed by electrophoresis in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and the catalytic activity was evaluated using lyophilized cells of Micrococcus luteus as substract. The antimicrobial activity was evaluated using specific methods for each enzyme. The sample toxicity was evaluated by cell viability using ATCC CCL-92 fibroblasts. The MdL1 obtained presented suitable homogeneity characteristics and activity of 108.35 U/mg. The HEWL has showed activity against S. aureus, M. luteus e C. albicans. MdL1 only showed activity against M. luteus. Due to the absence of antimicrobial activity of MdL1 in the evaluated concentration it was not submitted to the cytotoxicity test. Regarding HEWL, it has not showed citotoxicity in the previous test.

O aumento na procura por produtos naturais pelo mercado farmacêutico, cosmético e alimentício demanda pesquisas no desenvolvimento desses produtos. Esses são direcionados à obtenção de substâncias de origem vegetal ou animal, assim como, para produtos biotecnológicos. Investigações quanto à atividade antibacteriana de proteínas e peptídeos são realizadas. Dentre essas substancias, podemos citar as lisozimas, proteínas que hidrolisam as ligações &#946; 1-4 glicosídicas entre o ácido N-acetilmurâmico e N-acetilglicosamina, presentes no peptidoglicano da parede celular bacteriana. Além disso, apresentam atividade de quitinase, ou seja, quebram a ligação glicosídica da quitina presente na parede fúngica. As lisozimas apresentam alta especificidade pela parede microbiana indicando aparente ausência de efeitos tóxicos aos humanos. Assim, tornando-a candidata a agente antimicrobiano em formulações cosméticas e farmacêuticas. A lisozima de ovo de galinha tem atividade antimicrobiana, entretanto não havia estudos relacionados com os micro-organismos contaminantes normalmente encontrados em produtos farmacêuticos e cosméticos. Além disso, a lisozima recombinante de Musca domestica (MdL1) não possui ainda sua atividade antimicrobiana definida. Os objetivos do trabalho foram:1) Obtenção da lisozima recombinante de Musca domestica (MdL1); 2) Avaliação a atividade antimicrobiana da MdL1 e de lisozima de ovo de galinha, Hen Egg White Lysozyme (HEWL), frente à Staphylococcus aureus (ATCC 6538), Micrococcus luteus (ATCC 4698), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 8739), Candida albicans (ATCC 10231) e Aspergillus niger (ATCC 16404); 3) Avaliação da toxicidade da lisozima em cultura de células de fibroblastos (ATCC CCL-92). A MdL1 foi obtida por meio de expressão gênica em Pichia pastoris GS115 (Invitrogen), concentrada utilizando polietilenoglicol 6000 e dialisada contra água deionizada através da membrana com porosidade seletiva de 12kDa. A homogeneidade foi analisada por eletroforese em gel de poliacrilamida em condições desnaturantes; e a atividade catalítica foi avaliada utilizando células liofilizadas de Micrococcus luteus como substrato. A atividade antimicrobiana foi determinada por método específico para cada enzima. A toxicidade in vitro das amostras foi avaliada pela viabilidade celular de fibroblastos ATCC CCL-92. A MdL1 obtida apresentou características de homogeneidade adequadas e atividade de 108,35 U/mg. A HEWL mostrou-se ativa contra S. aureus, M. luteus e C. albicans. A MdL1 apresentou-se ativa contra M. luteus, apenas. Devido a ausência de atividade antimicrobiana a MdL1 não foi submetida a avaliação citotóxica. Em relação à HEWL não demonstrou citotoxicidade na avaliação prévia realizada. / The increase in the search for natural products by pharmaceutical, cosmetic and food markets requires researches in these products development. These are directed to obtaining substances of vegetal or animal origin, as well as biotechnological products. The research in relation to antibacterial activity of proteins and peptides is carried out. Among these substances, it is possible to mention the lysozyme, protein that catalyze the break of 1,4-beta-D glucosidic bond between N-acetylmuramic acid and N-acetilglicosamine which are present in peptidoglicane of the bacterial cell wall. Besides, there is kitinase activity, that is, they break the glicosidic bond of chitin which is present in fungal wall. The lysozymes show high specificity by microbial wall indicating apparent absence of toxicological effects to human beings. Therefore, it becomes the candidate to antimicrobial ingredient in cosmetic and pharmaceutical dosage forms. The hen egg white lysozyme has antimicrobial activity, however there were no studies related to spoiled microorganism usually found in pharmaceutical and cosmetic products. In addition, there were not studies about microbial activity of recombinant Musca domestica lysozyme 1 (MdL1). The aim of this research was: 1) To obtain MdL1; 2) Evaluation of MdL1 and hen egg white lysozyme (HEWL) antimicrobial activity against Staphylococcus aureus (ATCC 6538), Micrococcus luteus (ATCC 4698), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 8739), Candida albicans (ATCC 10231) e Aspergillus niger (ATCC 16404); 3) Evaluation of lysozyme toxicity in fibroblast cells culture (ATCC CCL-92). The MdL1 was expressed as recombinant protein in Pichia pastoris GS115 (Invitrogen), concentrated using polyethylene glycol 6000 and dialyzed against deionized water through the selective porosity of 12kDa membrane. The homogeneity was analyzed by electrophoresis in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and the catalytic activity was evaluated using lyophilized cells of Micrococcus luteus as substract. The antimicrobial activity was evaluated using specific methods for each enzyme. The sample toxicity was evaluated by cell viability using ATCC CCL-92 fibroblasts. The MdL1 obtained presented suitable homogeneity characteristics and activity of 108.35 U/mg. The HEWL has showed activity against S. aureus, M. luteus e C. albicans. MdL1 only showed activity against M. luteus. Due to the absence of antimicrobial activity of MdL1 in the evaluated concentration it was not submitted to the cytotoxicity test. Regarding HEWL, it has not showed citotoxicity in the previous test.