Orientador: Lucilene Delazari dos Santos / Resumo: O uso de animais de grande porte como sorodoadores tem se mostrado de grande importância para a produção do selante de fibrina do Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP). O objetivo deste trabalho foi certificar um plantel de bubalinos, realizar uma investigação sobre os potenciais candidatos à biomarcadores de brucelose em soro de búfalos e evidenciar o isolamento e a quantificação pioneira de exossomos (EVs) do soro de bubalinos com theileriose e babesiose. Realizaram-se os seguintes testes sorológicos: brucelose (BRU), leptospirose (LEP), febre aftosa (FMD), rinotraqueíte infecciosa bovina (IBR), diarréia viral bovina (BVD), varíola bovina (Pox) e tuberculose, língua azul (BTV), estomatite vesicular (EV) - sorotipos cocal (COCV) e alagoano (VSAV), leucose bovina (BLV), neospora (NC), toxoplasmose (TX) e testes bioquímicos laboratoriais. Todas as amostras de soro foram submetidas ao diagnóstico sorológico para detecção de brucelose, diagnóstico de reação em cadeia da polimerase para detecção de theileriose e babesiose, cromatografia líquida de afinidade, isolamento de exossomos, digestão de proteínas em solução, análise de espectrometria de massa e ferramentas de bioinformátca. Observamos um enriquecimento de 90,5% de proteínas nos soros de búfalos após este protocolo de depleção. A análise MS/MS evidenciou que as principais diferenças no proteoma dos animais com brucelose. No que se refere ao exossosmos isolados e quantififcados neste trabalho, nossa met... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The use of large animals as sorbozers has been shown to be of great importance for the production of fibrin sealant from the Center for the Study of Venoms and Poisonous Animals (CEVAP). The objective of this work was to certify a flock of buffaloes, to conduct an investigation of the potential candidates for brucellosis biomarkers in buffalo serum and to evidence the isolation and the pioneer quantification of buffalo serum exosomes (EVs) with theileriose and babesiosis. The following serological tests were carried out: brucellosis (BRU), leptospirosis (LEP), foot-and-mouth disease (FMD), infectious bovine rhinotracheitis (IBR), bovine viral diarrhea (BVD), bovine pox (Pox) and tuberculosis, blue tongue (BTV), vesicular stomatitis (EV) - cocal (COCV) and alagoan (VSAV) serotypes, bovine leukosis (BLV), neospora (NC), toxoplasmosis (TX) and laboratory biochemical tests. All serum samples were submitted: 1) serological test; 2) Polymerase chain reaction; 3) Depletion; 4) Isolation of exosomes; 5) Mass spectrometry and 6) Analysis and interpretation of the data. The MS / MS analysis showed that the major differences in the proteome of animals with brucellosis. Regarding the isolated and quantified exosmos in this work, our methodology is strongly encouraged for the isolation and characterization of EVs in Apicomplexa infections in farm animals. We conclude that this study on different fronts such as management, infectious diseases and parasitic diseases can be used in small pro... (Complete abstract click electronic access below) / Doutor

O objetivo deste trabalho foi analisar as mudanças no perfil protéico em películas adquiridas formadas in situ, em diferentes tempos, sobre o esmalte e sobre a dentina humana após a exposição a dois tipos de ácido: lático e cítrico. Foram utilizados 162 blocos de esmalte e 162 blocos de dentina humana (3X3 mm). Os experimentos foram realizados em três dias consecutivos. Em cada dia, os voluntários (n=9) utilizavam um dispositivo mandibular contendo 6 blocos de esmalte e 6 blocos de dentina. Na sequência, os voluntários permaneceram com o dispositivo na cavidade bucal por 10 ou 120 minutos para formação de película adquirida. Os blocos foram então imersos em ácido cítrico (1%, pH 2,5) ou ácido lático (0,1 M pH 4,8) ou água deionizada por 20 segundos. Na sequência, a película foi removida com um papel de filtro umedecido em ácido cítrico a 3%. Este procedimento foi repetido por mais dois dias adicionais e foi confeccionado um pool com os papeis de filtro obtidos dos 9 voluntários, para cada tipo de substrato, tempo de coleta e meio de imersão. Após extração das proteínas, as mesmas foram submetidas à cromatografia líquida de fase reversa interligada a um espectrômetro de massas (nLC-ESI-MS/MS). Os dados obtidos de MS/MS foram processados e submetidos conjuntamente ao programa SEQUEST [Proteome Discoverer 1.3 (Thermo Scientific)]. As buscas foram feitas utilizando-se os bancos de dados SWISS-PROT e TrEMBL. Para o esmalte, a taxa de identificação de proteínas foi baixa (13 proteínas no total). Já para a dentina, a taxa de identificação de proteínas foi maior (223 proteínas no total), sendo que a exposição ao ácido cítrico reduziu dramaticamente o número de proteínas identificado, o que não aconteceu para o ácido lático. Proteínas ácido-resistentes foram identificadas tanto para o esmalte quanto para a dentina, destacando-se as queratinas e as mucinas, respectivamente. Estas proteínas, ou os peptídeos oriundos delas responsáveis pelo efeito protetor, são candidatas a serem utilizadas para o enriquecimento de produtos odontológicos, visando à prevenção da cárie e erosão dentária. / The purpose of this work was to analyze changes in protein profile in the acquired pellicle formed on enamel and dentine at different times in situ after exposure to lactic and citric acids. Enamel (n=162) and dentin (n=162) blocks (3X3 mm) were be used. The experiments were conducted on three consecutive days. Each day, volunteers (n = 9) used a mandibular device containing 6 blocks of enamel and 6 of human dentin. After the volunteers remained with the device in the oral cavity for 10 or 120 minutes in order to allow the formation of the acquired pellicle, the blocks were immersed in citric acid (1%, pH 2.5) or lactic acid (0.1 M, pH 4,8) or deionized water for 20 seconds. Following, the pellicle was collected with an electrode filter paper soaked in 3% citric acid. This procedure was repeated for two additional days and \'pools\' with the filter papers obtained from the 9 volunteers for each type of substrate, sampling time and type of acid were made. After extraction, proteins were subjected to reverse phase liquid chromatography coupled to mass spectrometry (nLC-ESI-MS/MS). MS/MS data were processed and submitted to SEQUEST software [Proteome Discoverer 1.3 (Thermo Scientific)]. Searches were done using SWISS-PROT and TrEMBL databases. The rate of protein identification was low for enamel (13 proteins in total). As for dentin, the rte of protein identification was higher (223 proteins in total). Exposure to citric acid dramatically reduced the number of identified proteins, what did not occur for lactic acid. Acid-resistant proteins, especially keratins and mucins were identified for enamel and dentin, respectively. These proteins, or the peptides originated from them that are responsible for the protective effect, are candidates to be used for the enrichment of dental products, aiming to prevent dental caries and erosion.

Les stratégies de protéomique quantitative sans marquage sont très attractives dans le domaine de la recherche de biomarqueurs de pathologies. Cependant, elles requièrent une pleine maîtrise du schéma analytique et de sa répétabilité. Plus particulièrement, la préparation d’échantillons nécessite d’être suffisamment répétable pour ne pas impacter la qualité et la fiabilité des résultats. Les objectifs de cette thèse étaient de développer et d’optimiser des méthodes analytiques pour la protéomique quantitative, en particulier pour l’étape de préparation d’échantillons. Ainsi, un protocole innovant, simple, rapide et permettant l’analyse quantitative sans marquage d’un grand nombre d’échantillons avec une haute répétabilité a été développé et optimisé : le « Tube-Gel ». Par ailleurs, des préparations d’échantillons adaptées à différentes matrices biologiques pour la recherche de biomarqueurs ont été élaborées. Les méthodes mises au point et leur application ont permis de proposer des candidats biomarqueurs pour plusieurs pathologies : le glioblastome, les lymphomes B diffus à grandes cellules, et les complications survenant sur les greffons rénaux. / Label-free quantitative proteomics strategies are very attractive for diseases biomarkers researches. These approaches require the full control and the repeatability of the analytical workflow. In particular, the sample preparation has to be repeatable enough to ensure the quality and reliability of the results. Objectives of this work were to optimize and develop analytical methods for quantitative proteomics, with a special focus on the sample preparation step. Thus, an innovative, easy and fast protocol allowing the analysis of high sample numbers with high repeatability was developed and further optimized: the “Tube-Gel” protocol. Besides,sample preparations adapted to a variety of biological matrices were developed for the search of biomarkers. The developed methods and their application allowed the identification of potential biomarkers for a variety of diseases: glioblastoma, diffuse large B-cell lymphomas and renal transplants failures.

Submitted by Luciana Sebin (lusebin@ufscar.br) on 2016-09-19T12:55:56Z No. of bitstreams: 1 TeseHABSP.pdf: 4918857 bytes, checksum: fde7822ef1d8c23c0e0c60d9ccaf8152 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-20T18:13:37Z (GMT) No. of bitstreams: 1 TeseHABSP.pdf: 4918857 bytes, checksum: fde7822ef1d8c23c0e0c60d9ccaf8152 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-20T18:13:44Z (GMT) No. of bitstreams: 1 TeseHABSP.pdf: 4918857 bytes, checksum: fde7822ef1d8c23c0e0c60d9ccaf8152 (MD5) / Made available in DSpace on 2016-09-20T18:13:53Z (GMT). No. of bitstreams: 1 TeseHABSP.pdf: 4918857 bytes, checksum: fde7822ef1d8c23c0e0c60d9ccaf8152 (MD5) Previous issue date: 2015-12-18 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / In a previous study conducted by our group, it was noticed that fluoride (F) can induce changes in the expression of several liver proteins. Reports in the literature suggest that the changes caused by F in the body are dose- and time-dependent. The objective of this study was to analyze the effect of different F concentrations, exposure time to this ion and concomitant exposure to a high calorie diet in the metabolism of lipids and protein expression in the liver of rats. The study was divided into 2 steps. The first step included 72 21-day-old male Wistar rats that were divided into 2 groups (n=36) according to the diet administered (AIN-93M and Presence). Each group was further divided according to the duration of the treatment (20 or 60 days). In addition, each these was divided into 3 subgroups (n=6), according to the concentration of F administrated in the drinking water, as follows: 0 mg/L (control), 15 mg/L or 50 mg/L. After the experimental period, the animals were anesthetized and the liver and blood were collected. F analysis in plasma and liver tissue was done. Part of the liver was fixed for histological analysis. Lipids were analyzed in plasma and triglycerides were analyzed in the liver. Expressions of proteins were evaluated in the liver by Western blotting. In the second step the only Presence diet were use and the groups experimental the same as previously described. At the end of experimental period, liver and plasma were collected. F concentration in plasma and liver were analyzed. Liver proteins were extracted and prepared for mass spectrometry analysis. Proteins were sequenced and identified. The analysis of F concentrations indicated a doseresponse increase in plasma, regardless the time of exposure to F and type of diet. F concentrations in the liver were higher in the groups receiving 50 ppm F in respect to control. Administration of F altered the lipid profile, with a reduction in TGA in plasma and increase in HDL when the hypercaloric diet was used. The expression of GRP78, ERP29 and SOD2 and Apo-E was altered by F, under the influence of time and type of diet administered. For the groups receiving 50 mgF/L, it was observed an increase in the concentration of proteins related to the defense against oxidative stress and ER stress. For the group that received the concentration of 15 mgF/L, changes in structural proteins, mitochondrial proteins and proteins related to cell proliferation were observed, depending on the time of administration. The results suggest an adaptive mechanism of liver upon exposure to F, which seems to be related to the activation of proteins related to maintenance of homeostasis and that fight against oxidative stress and ER stress caused by this ion. / Em trabalho prévio realizado pelo nosso grupo foi observado que o fluoreto (F) pode provocar alterações na expressão de várias proteínas hepáticas. Relatos na literatura sugerem que as alterações causadas pelo F no organismo são dose e tempo-dependentes. Assim, o objetivo deste trabalho foi analisar o efeito da administração de diferentes concentrações de F, do tempo de exposição a este íon e da exposição concomitante a uma dieta hipercalórica no metabolismo de lipídios e expressão de proteínas hepáticas em ratos. O trabalho foi realizado em 2 etapas. Na primeira, foram utilizados 72 ratos Wistar machos com 21 dias, que foram divididos em 2 grupos (n=36) de acordo com o tipo de dieta (AIN-93M ou Presence), então subdividido em 2 grupos (n=18) de acordo com o tempo de tratamento (20 ou 60 dias). Cada grupo foi dividido em 3 subgrupos (n=6), de acordo com a dose de fluoreto a ser administrada através da água de beber, a saber: 0 mg/L, 15 mg/L ou 50 mg/L. Decorridos os períodos experimentais o fígado e o sangue foram coletados. Foi realizada a análise de F no plasma e tecido hepático. Parte do fígado foi fixado para a confecção das lâminas para análise histológica. No plasma foi realizada a análise de perfil lipídico e no fígado, de triglicerídeos. Foi avaliada a expressão das proteínas hepáticas por Western Blotting. Na segunda etapa foi realizada apenas com a ração presence com os mesmos grupos experimentais da primeira etapa. Ao final do período experimental, o fígado e o plasma foram coletados. A concentração de F no plasma e no fígado foram analisada. Foi realizada a extração de proteínas e preparação das proteínas do fígado para espectrometria de massa, sendo então sequenciadas e identificadas. A análise das concentrações de F indicaram um aumento dose-resposta no plasma, independente do período administrado ou tipo de dieta. Já as concentrações de F no fígado foram maiores nos grupos que receberam 50 mg/L de F em relação ao controle. A administração de F alterou o perfil lipídico, com uma redução no TGA no plasma e aumento do HDL. As inclusões lipídicas no fígado foram reduzidas no grupo que recebeu 50 ppm F por 20 dias em conjunto com a dieta hipercalórica. A expressão da GRP78, ERP29, SOD2 e Apo-E foram alteradas pelo F, sob influência do tempo e dieta administrada. Para os grupos que receberam a concentração de 50 ppm F foi observado um aumento na concentração de proteínas relacionadas à a defesa contra o estresse oxidativo e do RE. Para o grupo que recebeu a concentração de 15 ppm F houve alterações estruturais, mitocondriais e relacionadas à proliferação, verificando-se um efeito depende do tempo. Desta forma, podemos concluir que o fígado possui um mecanismo de adaptação à ação do F e que o mesmo parece estar relacionado ao acionamento de proteínas referentes à homeostasia e contra o estresse oxidativo e do RE provocado por este íon. / FAPESP: 2011/17263-9

The thesis "The proteomic analysis in hematology: Identification of alfa2- macroglobulin as a specific carrier for the hormone hepcidin and proteomic analysis of the leukemic K562 cell differentiation induced by sodium butyrate" describes proteomic approaches, used for the identification and functional characterisation of proteins, which are binding and transporting the iron metabolism regulating hormone hepcidin. Proteomic techniques are also exploited for the identification of proteins, participating in erythroid differentiation of the model cell line K562. In the first section of the thesis, non-denaturing, native techniques, such as chromatography and native electrophoresis are used, in the second section, the control and butyrate - induced K562 cell proteomes are compared using the classical 2D - SDS polyacrylamide gel electrophoresis approach. The methods, described in the thesis are broadening the spectrum of available techniques in experimental hematology. The results, described in this thesis together with the accompanying published manuscripts broaden our knowledge in the function of proteins of iron metabolism and proteins, functioning in erythroid differentiation. Key words: proteomic analysis, hepcidin, alfa2-macroglobulin, iron metabolism, CML, K562, sodium butyrate

O objetivo deste trabalho foi analisar as mudanças no perfil protéico em películas adquiridas formadas in situ, em diferentes tempos, sobre o esmalte e sobre a dentina humana após a exposição a dois tipos de ácido: lático e cítrico. Foram utilizados 162 blocos de esmalte e 162 blocos de dentina humana (3X3 mm). Os experimentos foram realizados em três dias consecutivos. Em cada dia, os voluntários (n=9) utilizavam um dispositivo mandibular contendo 6 blocos de esmalte e 6 blocos de dentina. Na sequência, os voluntários permaneceram com o dispositivo na cavidade bucal por 10 ou 120 minutos para formação de película adquirida. Os blocos foram então imersos em ácido cítrico (1%, pH 2,5) ou ácido lático (0,1 M pH 4,8) ou água deionizada por 20 segundos. Na sequência, a película foi removida com um papel de filtro umedecido em ácido cítrico a 3%. Este procedimento foi repetido por mais dois dias adicionais e foi confeccionado um pool com os papeis de filtro obtidos dos 9 voluntários, para cada tipo de substrato, tempo de coleta e meio de imersão. Após extração das proteínas, as mesmas foram submetidas à cromatografia líquida de fase reversa interligada a um espectrômetro de massas (nLC-ESI-MS/MS). Os dados obtidos de MS/MS foram processados e submetidos conjuntamente ao programa SEQUEST [Proteome Discoverer 1.3 (Thermo Scientific)]. As buscas foram feitas utilizando-se os bancos de dados SWISS-PROT e TrEMBL. Para o esmalte, a taxa de identificação de proteínas foi baixa (13 proteínas no total). Já para a dentina, a taxa de identificação de proteínas foi maior (223 proteínas no total), sendo que a exposição ao ácido cítrico reduziu dramaticamente o número de proteínas identificado, o que não aconteceu para o ácido lático. Proteínas ácido-resistentes foram identificadas tanto para o esmalte quanto para a dentina, destacando-se as queratinas e as mucinas, respectivamente. Estas proteínas, ou os peptídeos oriundos delas responsáveis pelo efeito protetor, são candidatas a serem utilizadas para o enriquecimento de produtos odontológicos, visando à prevenção da cárie e erosão dentária. / The purpose of this work was to analyze changes in protein profile in the acquired pellicle formed on enamel and dentine at different times in situ after exposure to lactic and citric acids. Enamel (n=162) and dentin (n=162) blocks (3X3 mm) were be used. The experiments were conducted on three consecutive days. Each day, volunteers (n = 9) used a mandibular device containing 6 blocks of enamel and 6 of human dentin. After the volunteers remained with the device in the oral cavity for 10 or 120 minutes in order to allow the formation of the acquired pellicle, the blocks were immersed in citric acid (1%, pH 2.5) or lactic acid (0.1 M, pH 4,8) or deionized water for 20 seconds. Following, the pellicle was collected with an electrode filter paper soaked in 3% citric acid. This procedure was repeated for two additional days and \'pools\' with the filter papers obtained from the 9 volunteers for each type of substrate, sampling time and type of acid were made. After extraction, proteins were subjected to reverse phase liquid chromatography coupled to mass spectrometry (nLC-ESI-MS/MS). MS/MS data were processed and submitted to SEQUEST software [Proteome Discoverer 1.3 (Thermo Scientific)]. Searches were done using SWISS-PROT and TrEMBL databases. The rate of protein identification was low for enamel (13 proteins in total). As for dentin, the rte of protein identification was higher (223 proteins in total). Exposure to citric acid dramatically reduced the number of identified proteins, what did not occur for lactic acid. Acid-resistant proteins, especially keratins and mucins were identified for enamel and dentin, respectively. These proteins, or the peptides originated from them that are responsible for the protective effect, are candidates to be used for the enrichment of dental products, aiming to prevent dental caries and erosion.

Proteomic techniques were used to evaluate the protein profile of the earthworm, (Lumbricus terrestris), following a bacterial challenge. One control group received no injection; a second control group received injections of phosphate buffer solution (PBS). The experimental group received injections of PBS containing (Aeromonas hydrophila). After incubation for 12 hours at 20°C, coelomic fluid was collected from each group for analysis by 2-D electrophoresis. There were significant differences in spot appearance and density between control and experimental groups. Sixteen spots showed a two-fold increase in density and 63 showed at least a two-fold decrease in density between samples from control and bacteria-challenged earthworms, respectively, suggesting up- and down-modulation of proteins potentially involved in the earthworm's response to bacterial challenge.

La sicurezza e la qualità degli alimenti sono tutt’ora un problema critico per i paesi in via di sviluppo. Le diete a basso contenuto di acido folico, per esempio, possono causare gravi problemi di salute, soprattutto nei bambini. Gravi disturbi legati al tubo neurale (DTN) nei neonati possono derivare infatti da madri che hanno insufficiente apporto di acido folico (400-600 g / giorno) durante il periodo di gravidanza. Inoltre, se non adeguatamente protetti o trattati, I prodotti alimentari possono essere vettori di funghi e batteri patogeni rappresentando una fonte potenziale di malattie per l’uomo e una perdita economica per le industrie agro-alimentari. Nella seguente tesi si è quindi quindi studiato il ruolo di batteri lattici selezionati (LAB) in grado di aumentare il valore nutrizionale del latte attraverso la produzione di acido folico durante il processo di fermentazione. Inoltre, ci si è concentrati sul loro uso come "bio-conservanti" contro funghi e batteri, attraverso la sintesi di composti antimicrobici (batteriocine) in grado di inibire la crescita di funghi filamentosi e/o batteri patogeni. / The safety and quality of food are still a critical issue in developing countries. Diets with a low content of folic acid, for example, may cause serious health problems, especially in children. Severe disorders related to neural tube (NTD) in infants may arise from mothers having inadequate intakes of folic acid (400-600 g/dia) during the mother pregnancy period. Moreover foods, when not properly protected or treated, can be vectors of pathogenic fungi and bacteria thereby representing a potential source of human diseases and an economical loss for the food industry. In the following thesis we have therefore investigated the role of selected lactic acid bacteria (LAB) in increasing the nutritional value of milk through the production of folic acid during the fermentation process. In addition, we focused on their use as “bio-preservatives” against fungal and bacterial spoilage, through the synthesis of antimicrobial compounds (bacteriocins) able to inhibit the growth of filamentous fungi and /or pathogenic bacteria.

This diploma thesis was focused on the study of lysozyme and lysozyme-like proteins, either of similar function (antibacterial) or molecular weight (14 - 17 kDa), of synanthropic acaroid mites. In general, animals utilize lysozymes for defensive (antimicrobial) or digestive purposes but also as a digestive enzyme. Some chitinases or other enzymes that act similarly to lysozyme can be utilized for similar purposes. Chitinases belong to house dust mite allergens. One of major mite are historically named lysozyme-like proteins which name relates to their size similar to lysozyme. Bacteriolytic activity has also 14.5 kDa (UniprotKB Q8MWR6) protein. The species selected for the study were domestic mites Dermatophagoides farinae, D. pteronyssinus and Lepidoglyphus destructor. Presence of lysozyme was detected by direct detection with polyclonal antibody using immunohistochemistry and dot blots. Immunohistochemistry proved presence of lysozyme epitopes in the feces of D. farinae, D pteronyssinus a L. destructor. Dot blot analysis demonstrated the presence of imunoreactivity of antibody in spent growth medium extracts (SGME) of all three species. This implies that lysozyme is synthesized in the midgut. The presence of lysozyme and lysozyme-like proteins was proved using 2D electrophoresis and MALDI TOF/TOF...

L’amélioration des performances de reproduction des poissons d’élevage nécessite de déterminer les facteurs intrinsèques et extrinsèques influençant la qualité des gamètes d’une part, et de définir des paramètres fiables permettant de prédire les performances de reproduction d’autre part. Notre objectif est donc de comprendre le déterminisme multifactoriel de la reproduction chez la perche commune, Perca fluviatilis. Quatre facteurs nutritionnels (type d’aliment et taux de rationnement distribués lors des phases d’induction et de vernalisation) et 3 facteurs populationnels (poids initial, origine géographique, niveau de domestication) ont été testés. Une différence de réponse entre les sexes a été observée. Le type d’aliment distribué en vernalisation et le poids initial ont modifié l’état général des femelles. Les mâles ont plutôt été sensibles aux taux de rationnement et à l’origine géographique. L’étude des performances de reproduction a montré que le taux de ponte était sous l’influence de l’interaction entre le type d’aliment distribué en induction et en vernalisation, tandis que l’origine géographique a modulé la date de ponte. La régulation des performances de reproduction est donc un mécanisme complexe sous l’influence simultanée de plusieurs facteurs. La seconde partie de ce travail concerne la recherche de marqueurs prédictifs de la qualité des ovules. Nous avons d’abord montré que peu de paramètres morpho-anatomiques des pontes ou ovules sont des prédicateurs fiables. Cependant, l’analyse protéomique a permis de mettre en évidence plusieurs protéines exprimées différemment selon la qualité des pontes, pouvant jouer le rôle de biomarqueurs de qualité des ovules / Improving fish reproduction in breeding conditions implies to understand intrinsic and extrinsic factors influencing gametes quality on the one hand and to define relevant parameters allowing the prediction of fish reproductive performance on the other hand. Our goal was thus to understand the multifactorial determinism of the common perch (Perca fluviatilis) reproduction. Four nutritional factors (type of food and rate of rationing used either during the induction or vernalization phases) and 3 populational factors (initial weight, geographic origin and domestication level of breeders) have been tested. Data show different responses between females and males. type of food during wintering phase and initial broodstock weigh modified female condition. Males have been sensitive to rationing during wintering phase as well as geographical origin. Data show also that spawning rate was under the influence of interaction between kind of food during wintering phase and induction whereas geographical origin modulated the spawning date. The regulation of the performance reproduction is also a complex mechanism influenced by several factors. The second part of this work consisted on the research of parameters potentially predictive of ova quality. Firstly, our work shows that morphometric parameters measured before the fertilization are poorly relevant to predict reproductive performance. However, the proteomic analysis of several spawn allowed us to highlight proteins differently expressed according to the spawn quality, such proteins could be ova quality biomarkers