Research Doctorate - Doctor of Philosophy (PhD) / Membrane fusion events are a fundamental aspect of cellular biology and underpin important processes such as organ formation and fertilisation. Within the latter, proteins that are expressed on the egg surface which are responsible for mediating sperm recognition, binding and fusion to the egg, are yet to be fully determined. Evidence does however suggest that egg surface glycophosphatidylinositol (GPI)-anchored proteins play a role in sperm binding, whilst another class of proteins, known as tetraspanins, appear to be important in downstream events of membrane fusion. Of the tetraspanins, CD9 and CD81 have been identified as fulfilling roles in membrane fusion; identifications are however yet to obtained for the important GPI-anchored protein(s). This research aimed to identify and characterise egg surface proteins implicated in sperm-egg interaction, and embodied attempts to both identify the important GPI-anchored protein(s) as well as expand upon tetraspanin studies through investigations into mice lacking the tetraspanin CD151. Throughout this research, it was hypothesised that membrane fusion events of fertilisation parallelled those of enveloped virus – host cell fusion, for which rearrangement of surface protein thiols is essential. In vitro binding and fusion experiments were utilised as functional bioassays in the investigation of factors affecting sperm-egg interaction, such as tetraspanin deletion and the xenobiotic modification of cell surface thiols, while mass spectrometry (MS)-based proteomics and bioinformatics-based analyses were employed to compile oocyte protein databases and to identify candidate proteins responsible for mediating sperm-egg interaction, such as GPI-anchored proteins. It was determined that exposing oocytes to compounds with a capacity to alkylate cell surface thiols strongly inhibited sperm-egg binding. Additionally, while CD151 deletion had no effect on sperm-egg binding, the downstream events of membrane fusion were significantly impaired. Ovaries from CD151 null mice also exhibited abnormal phenotypes. In addition, a total of 11 identifications were obtained in the search for the GPI-anchored proteins expressed within eggs, however only 6 of these were deemed to be potential mediators of sperm-egg interaction. In conclusion, the experiments outlined herein demonstrate a novel inhibitory effect for specific xenobiotics on sperm-egg interaction, and correlate the inhibitory action of these compounds with their capacity to reduce cell surface thiol labelling. A novel role for CD151 in the mediation of sperm-egg fusion was also discovered, while at the same time the important GPI-anchored protein(s) implicated in sperm-egg binding may be among 6 identified potential candidates. Together the findings reiterate the consensus that oocytes possess a cell surface protein complex responsible for mediating sperm binding and fusion as separate events, and in light of the demonstrated importance of surface thiols, that events of sperm-egg membrane fusion parallel those of enveloped virus – host cell fusion.

This study investigates the reproductive strategies of the pill-box crab, Halicarcinus cookii on the Kaikoura Peninsula, New Zealand. Various aspects essential to understanding reproductive strategies were examined including growth, population dynamics, reproductive biology and mating behaviour. H. cookii exhibits obvious sexual dimorphism such that females develop wide abdomens forming brood chambers, and males tend to grow larger than females and have larger chelipeds in relation to body size. H. cookii allocates energy into growth and reproduction in separate phases of its life cycle where growth ceases as reproductive maturity begins due to a terminal/pubertal moult. Despite the presence of ovigerous females throughout the 15 month sampling period, the population was highly seasonal, with peaks in recruitment and growth occurring primarily during the winter months and peaks in numbers of mature individuals during the summer months. Reproductive output increased with body size in H. cookii, as larger females produced more eggs and larger males transferred more sperm than their smaller counterparts. Ovaries matured prior to the terminal/pubertal moult (anecdysis) and, in multiparous females, in synchrony with brood development, allowing females to produce broods in quick succession, maximising their reproductive output in their short life span (approximately 12-18 months, 6 months as an adult). Incubation duration of broods decreased as seawater temperature increased, suggesting that temperature is the primary cause of the seasonal population cycling. Sperm storage allowed females to produce at least 4 fertilised broods without re-mating. Some sperm mixing in the spermathecae appeared to occur and the ventral-type structure implies last male sperm precedence. Males therefore preferentially mated with females closest to laying a new brood and guarded them longer than other females to ensure their paternity. Guarding duration varied according to the sex ratio allowing males to maximise their reproductive output.

Research Doctorate - Doctor of Philosophy (PhD) / Membrane fusion events are a fundamental aspect of cellular biology and underpin important processes such as organ formation and fertilisation. Within the latter, proteins that are expressed on the egg surface which are responsible for mediating sperm recognition, binding and fusion to the egg, are yet to be fully determined. Evidence does however suggest that egg surface glycophosphatidylinositol (GPI)-anchored proteins play a role in sperm binding, whilst another class of proteins, known as tetraspanins, appear to be important in downstream events of membrane fusion. Of the tetraspanins, CD9 and CD81 have been identified as fulfilling roles in membrane fusion; identifications are however yet to obtained for the important GPI-anchored protein(s). This research aimed to identify and characterise egg surface proteins implicated in sperm-egg interaction, and embodied attempts to both identify the important GPI-anchored protein(s) as well as expand upon tetraspanin studies through investigations into mice lacking the tetraspanin CD151. Throughout this research, it was hypothesised that membrane fusion events of fertilisation parallelled those of enveloped virus – host cell fusion, for which rearrangement of surface protein thiols is essential. In vitro binding and fusion experiments were utilised as functional bioassays in the investigation of factors affecting sperm-egg interaction, such as tetraspanin deletion and the xenobiotic modification of cell surface thiols, while mass spectrometry (MS)-based proteomics and bioinformatics-based analyses were employed to compile oocyte protein databases and to identify candidate proteins responsible for mediating sperm-egg interaction, such as GPI-anchored proteins. It was determined that exposing oocytes to compounds with a capacity to alkylate cell surface thiols strongly inhibited sperm-egg binding. Additionally, while CD151 deletion had no effect on sperm-egg binding, the downstream events of membrane fusion were significantly impaired. Ovaries from CD151 null mice also exhibited abnormal phenotypes. In addition, a total of 11 identifications were obtained in the search for the GPI-anchored proteins expressed within eggs, however only 6 of these were deemed to be potential mediators of sperm-egg interaction. In conclusion, the experiments outlined herein demonstrate a novel inhibitory effect for specific xenobiotics on sperm-egg interaction, and correlate the inhibitory action of these compounds with their capacity to reduce cell surface thiol labelling. A novel role for CD151 in the mediation of sperm-egg fusion was also discovered, while at the same time the important GPI-anchored protein(s) implicated in sperm-egg binding may be among 6 identified potential candidates. Together the findings reiterate the consensus that oocytes possess a cell surface protein complex responsible for mediating sperm binding and fusion as separate events, and in light of the demonstrated importance of surface thiols, that events of sperm-egg membrane fusion parallel those of enveloped virus – host cell fusion.

Diss. (sammanfattning) Skara : Sveriges lantbruksuniv., 2002. / Härtill 4 uppsatser.

Diss. (sammanfattning). Swedish University of Agricultural Sciences : Sveriges lantbruksuniv. / Härtill 5 uppsatser.

Techn. Universiẗat, Diss., 2003--Braunschweig.

Made available in DSpace on 2014-06-11T19:32:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-08-05Bitstream added on 2014-06-13T19:43:51Z : No. of bitstreams: 1 pelicia_k_dr_botfmvz.pdf: 374037 bytes, checksum: b680d9b7127562753fb9bb889bfcf807 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O experimento teve por objetivo estudar o efeito de níveis de cálcio e da granulometria do calcário sobre o desempenho, qualidade de ovos e teor de cálcio no sangue e fezes de poedeiras semipesadas. As aves foram distribuídas em um delineamento fatorial inteiramente casualisado (3x4) com 12 tratamentos com quatro níveis de cálcio (3,0; 3,5; 4,0 e 4,5 %) e três composições granulmétricas do calcário: 100 % fino (0,18 mm); 50 % fino (0,18 mm) e 50 % grosso (3,13 mm) e 30 % fino (0,18 mm) e 70 % grosso (3,13 mm), seis repetições de oito aves por unidade experimental da linhagem Hisex Brown® de 59 a 70 semanas de idade. Não se observaram diferenças entre os tratamentos nem interações entre os fatores para percentagem postura, peso médio dos ovos, massa de ovos, consumo de ração conversão alimentar por dúzia e por quilograma de ovos produzidos, percentagem de ovos quebrados, percentagem de ovos inteiros, gravidade específica, espessura de casca, percentagem de casca, peso da casca por superfície de área, percentagem de gema, coloração da gema, % de albume, unidade Haugh e quantidade de cálcio no sangue. Observou-se efeito quadrático da composição granulométrica de calcário para percentagem de ovos defeituosos, com menor percentagem de ovos defeituosos usando composições granulométricas do calcário formado de 61,75 % de calcário de granulometria fina e 38,25 % de calcário de granulometria grossa. Houve efeito linear da equação de regressão do nível de cálcio da dieta para peso da casca por superfície de área e cálcio nas fezes onde a elevação no nível cálcio dietético aumentou o peso da casca por superfície de área e a percentagem de cálcio excretado. Conclui-se que a combinação do maior nível de cálcio (4,5%) na dieta com substituição do calcário fino pelo grosso em 38 % na alimentação de poedeiras... / The experiment was randomized with the objective to study the effect of the calcium levels and the limestone particle size on the performance, eggs quality and content of calcium in the blood and excrements in Brown laying hens. The birds had been distributed in an factorial delineation entirely randomized (3X4) with 12 treatments with four calcium levels (3.0, 3.5, 4.0 and 4.5 %) and three limestone particicle size compositions: 100 % thin (0.18 mm); 50 % thin (0.18 mm) and 50 % coarse (3.13 mm) and 30% thin (0.18 mm) and 70 % coarse (3.13 mm), six repetitions for treatment and eight birds for experimental unit of the Hisex Brown® lineage of 59 the 70 weeks of age. There were no differences (p>.05) for the treatments and nor significant interaction between the factors for posture percentage, average weight egg, egg mass produced, feed intake, feed conversion for dozen and kilogram of produced eggs, broken egg percentage, perfect egg percentage, specific gravity, shell thickness, eggshell and eight birds for experimental unit of the Hisex Brown® lineage of 59 the 70 weeks of age. There were no differences (p>.05) for the treatments and nor significant interaction between the factors for posture percentage, average weight egg, egg mass produced, feed intake, feed conversion for dozen and kilogram of produced eggs, broken... (Complete abstract click electronic access below)

Orientador: Antonio Carlos de Laurentiz / Resumo: O experimento foi conduzido na Universidade Estadual Paulista – UNESP, Faculdade de Engenharia de Ilha Solteira no Setor de Avicultura, com a finalidade de avaliar o efeito da valorização energética da goma de soja na dieta de poedeiras comerciais e verificar a viabilidade econômica de sua utilização como um produto comercial derivado da soja. Foram utilizadas 240 poedeiras da linhagem Lohmann LSL, com 49 semanas, durante um período de 112 dias (quatro ciclo de 28 dias) distribuídas em um delineamento inteiramente casualizado em um esquema fatorial (4x2) sendo esses: quatro níveis de energia metabolizável (2.610, 2.690, 2.770 e 2850 kcal/kg de ração) x dois níveis de inclusão de goma na dieta (0 e 2%), com cinco repetições de seis poedeiras cada. A goma de soja aumentou o consumo de ração, auxiliou no aumento do peso do ovo, da massa do ovo e do tamanho da gema do ovo, melhorou a digestibilidade do Extrato etéreo e da energia da ração. O índice Economic Feed Efficiency apontou melhor retorno econômico com o tratamento de menor nível de energia (2610 kcal/kg) independente da inclusão da goma. / Mestre

Orientadora: Isabel Cristina Boleli / Banca: Vera Maria Barbosa de Moraes / Banca: Maria Estela Gaglianone Moro / Resumo: O objetivo do presente estudo foi avaliar se a forma física da ração (farelada e peletizada) afeta a porosidade e espessura da casca, e porcentagem de perda de água e eclodibilidade dos ovos férteis de ovos de perdiz. Os ovos foram coletados logo após a postura e separados em diferentes intervalos de peso (35-45, 46-50, 51-55, 56-60 e 61-70g), sendo 10 ovos/classe/ração. O número de poros das cascas foi avaliado nas regiões apical, equatorial e basal dos ovos. Utilizou-se um micrômetro digital para a mensuração da espessura das três regiões da casca dos ovos, a partir dos quais obteve-se a espessura média por ovo. Os ovos para avaliação da eclodibilidade dos ovos férteis e porcentagem de perda de água do ovo foram pesados, desinfectados, incubados (T=35,5o C e UR= 60%) e transferidos no 16º dia de incubação para o nascedouro, onde foram mantidos até a eclosão. Os dados mostraram que, em perdizes, o número de poros na casca pode variar com a região da casca, peso do ovo e a forma física da ração, e que a ingestão de ração peletizada promove um aumento na espessura da casca dos ovos sem alterar a eclodibilidade dos ovos férteis e a porcentagem de perda de água dos mesmos durante a incubação. / Abstract: The aim of this study was to determine the effect of the ingestion of meal and pelleted diets on eggshell porosity and thickness, water loss and hatchability percentages of partridge (Rhynchotus rufescens) eggs. The eggs (n=100) were divided into five different weight classes (1: 35-45 g, 2: 46-50 g, 3: 51-55 g, 4: 56-60 g, 5: 61-70 g). Parts from different regions (air space, equator and small end) of each eggshell were analized in order to count pore numbers. Eggshell thickness measurements were carried out after removal of the egg membranes. The eggs used in the analyses of the water loss and hatchability percentages were weighed, desinfected, and then incubated (T=35.5oC and RH=60%). On the 16th day of incubation, these eggs were transferred to a hatchery maintained at temperature and RH similar to those used in the incubator, where they were kept until hatching. The results showed that the mean number of eggshell pores changed among the three regions of the eggshell and egg weight classes, and also between the physical form of diets. Ingestion of pelleted diet increased eggshell thickness; however, hatchability and water loss percentages remained unchanged. / Mestre

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-09-24T17:51:40Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_marques_bittencourt.pdf: 1215751 bytes, checksum: c3aad8d702755f268d177c8cafafe365 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-10-01T18:31:44Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_marques_bittencourt.pdf: 1215751 bytes, checksum: c3aad8d702755f268d177c8cafafe365 (MD5) / Made available in DSpace on 2018-10-01T18:31:44Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) tatiana_marques_bittencourt.pdf: 1215751 bytes, checksum: c3aad8d702755f268d177c8cafafe365 (MD5) Previous issue date: 2018 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O gr?o seco de destilaria com sol?veis (DDGS) ? um coproduto da ind?stria de etanol do milho e pode ser utilizado na alimenta??o animal, como substituto do farelo de soja. Foram realizados dois experimentos com o objetivo de avaliar diferentes n?veis de inclus?o de DDGS de milho na dieta de codornas e galinhas poedeiras, sobre o desempenho, qualidade dos ovos, pigmenta??o da gema e an?lise econ?mica. No experimento 1, foram utilizadas 210 codornas poedeiras (Coturnix japonica) com 23 a 31 semanas de vida, distribu?das em delineamento inteiramente casualizado com cinco tratamentos e seis repeti??es, com sete aves por unidade experimental. No experimento 2, foram 150 galinhas poedeiras (Hisex Brown) com 54 a 62 semanas de vida, distribu?das em delineamento interiramente casualizado com cinco tratamentos e seis repeti??es, com cinco aves por unidade experimental. Os tratamentos utilizados nos dois experimentos foram: ra??o controle; inclus?o de 5% de DDGS; inclus?o de 10% de DDGS; inclus?o de 15% de DDGS e inclus?o de 20% de DDGS. As vari?veis avaliadas foram: consumo de ra??o, convers?o alimentar por d?zia e massa de ovos, produ??o de ovos, peso dos ovos, peso da gema, alb?men, varia??o do peso corporal, viabilidade das aves, gravidade espec?fica, colora??o da gema, componente do ovo e an?lise econ?mica. No experimento 1, para as vari?veis consumo individual, convers?o alimentar por massa de ovos, taxa de postura, gravidade espec?fica e e colora??o da gema pelo leque colorim?trico houve efeito com a inclus?o do DDGS (P<0,05), para as demais vari?veis n?o houve efeito significativo. No experimento 2, para as vari?veis colora??o da gema pelo leque colorim?trico e colorimetro digital (L* e a*) houve efeito linear com a inclus?o do DDGS (P<0,05), para as demais vari?veis n?o foi observada diferen?a significativa. Quanto ? an?lise econ?mica, o DDGS pode ser considerado um alimento alternativo pelo baixo custo (R$). O DDGS de milho tem o potencial pigmentante na gema do ovo, fazendo com que apresente mais amarela sem influenciar o desempenho e a qualidade dos ovos das galinhas e codornas poedeiras, al?m de ser um produto mais econ?mico. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Zootecnia, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2018. / Dry distillers grain with Solubles (DDGS) is a Coproduct of ethanol corn industry and can be used in animal nutrition, as a substitute for soybean meal. Two experiments were conducted to evaluate different levels of inclusion of DDGS of cornmaize in the diet of laying hens and Japanese quail, about the performance, eggs quality, yolk pigmentation and economic analysis. In experiment 1, 210 were used laying quail (Coturnix japonica) with 23 to 31 weeks of life, distributed in completely randomized design with six treatments and five replicates, with seven quail per experimental unit. In experiment 2, were 150 laying hens (Hisex Brown) with 54 to 62 weeks of life, distributed in entirely randomized design with six treatments and five replicates, with five birds per experimental unit. The treatments used in two experiments were: T1: ration control; T2: inclusion of 5% of DDGS; T3: inclusion of 10% of DDGS; T4:15% inclusion of DDGS; T5: inclusion of 20% of DDGS. The variables evaluated were: feed intake, feed conversion per dozen and mass of eggs, egg production, egg weight, weight variation, viability of birds, gravity specifies, coloring of the yolk, egg component and economic analysis. The data were subjected to analysis by linear and quadratic regression models, the 5% probability. In experiment 1, to the individual consumption and feed conversion ratio by mass of eggs there was quadratic effect with the inclusion of DDGS (P < 0.05), for gravity and egg weight, egg yolk, and gem coloration for range and colorimetric digital colorimeter (b *) effect with the inclusion of DDGS (P < 0.05), for the other variables had no significant difference. In experiment 2, for the variables weight of yolk, albumen and bark, and gem coloration for range and colorimetric digital colorimeter (L *) there was no linear effect with the inclusion of DDGS (P < 0.05), the other variables had no significant difference. As the economic analysis the DDGS can be considered an alternative food for low cost (R$). The DDGS of corn has the potential pigmentante in egg yolk, making present more yellow mainly to higher levels, without affecting the performance and eggs quality of hens and laying quails, as well as being a more economic product.