Expanded bed chromatography using ion-exchange media is a powerful first step in purification processes. Expanded bed chromatography can be used to extract components from complex and viscous solution. This can be achieved because of the void created between adsorbent particles where as in packed bed chromatography, the adsorbent is too compact and dense for a complex feed stock to flow through. Expanded bed chromatography was used to purify bovine serum albumin (BSA) from chicken egg white (CEW). The high viscosity of CEW presents a unique challenge for efficient large-scale protein purification. This project aimed to optimize and evaluate a separation method that is believed to be particularly suitable for high viscosity solutions: expanded-bed ion exchange chromatography. The BSA was admixed into the CEW and the solution was pumped through the column for purification. The media used in the column was Streamline DEAE which is an anion-exchanger. The yield obtained was 85% and the purity was 57%. A mathematical model to understand and predict the behavior of expanded bed chromatography was developed to provide an estimation of the breakthrough curves obtained for BSA. A small sized porous dense adsorbent was also synthesized to enhance the purification process. This zirconia-based adsorbent allows use of higher flow velocities that is a key factor when working with viscous fluids such as chicken egg white.

Dietary intake is one of the major routes of human exposure to perfluoroalkyl and/or polyfluoroalkyl substances (PFAS). The objective of this study was to measure perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA) and perfluorohexane sulfonic acid (PFHxS) in organic and conventional egg from Sweden (n=8, consisted of 4 pooled eggs and 4 individuals) and China (n=9, consisted of 4 pooled eggs and 5 individuals) and compare the concentrations of PFAS between the two categories (organic and conventional). Also, to evaluate if there was any difference in concentrations of PFAS between both countries. In the end, evaluation of tolerable weekly intake of PFOS and PFOA due to consumption of egg recommended by the European Food Safety Authority (EFSA) was conducted if consuming these eggs would cause any human health risk. Liquid chromatography-mass spectrometry (LC-MS/MS) was used to analyze PFOS, PFOA and PFHxS in the egg samples. In egg samples from China, PFOA was the predominant PFAS; in an organic egg sample from Shenzhen with concentration up to 2000 pg/g, making up to 86% of the 3 PFAS. In contrast, PFOS had the greatest concentration of all PFAS in egg samples from Sweden and was detected in organic egg sample with concentration up to 184 pg/g, making up to 78% of the 3 PFAS. PFOA in samples from China was 18 times higher compared to egg samples to Sweden; results showed no significant differences in PFAS concentrations in egg samples between Sweden and China. In samples from China, concentrations of PFAS had total mean of 50 pg/g for PFOS, 373 pg/g for PFOA and 13 pg/g for PFHxS. In Sweden, mean concentrations of PFOS, PFOA, and PFHxS were found to be 5, 2, and 1,5 times (respectively) higher in organic eggs when compared to conventional. However, significant difference was only observed for PFOS in Swedish organic eggs (p<0.05, t-7.96, df=6). The different concentrations of contamination between organic and conventional egg could be due to the fish powder in organic chicken feed and ingestion of soil through pecking. The result suggests that current concentrations of PFOS and PFOA in organic and conventional chicken eggs are unlikely to cause any immediate harm to Swedish populations. For Chinese population since the consumption of egg has a high risk of exceeding the TWI, the current concentration of PFOA in organic chicken eggs may cause harm to the population based on TWIs established by EFSA. Further investigation is needed with more samples to be analyzed to confirm this point.

Post-weaning diarrhea is a major health challenge in the swine industry and is routinely managed by fortifying pig starter diets with antimicrobials. But there are concerns about antibiotic resistance, hence the need for identifying effective alternatives. The use of spray-dried whole egg powder containing anti-F4 antibodies (SDWE) against recombinant F4 antigens and chitosan oligosaccharide and Enterococcus fecalis probiotic combination (CPRO) was investigated in two trials using enterotoxigenic Escherichia coli K88+ (ETEC) oral challenge model in 21-d-old piglets. Pre-challenge, SDWE supported higher (P < 0.05) piglet performance whereas during the post-challenge period, SDWE and CPRO had no effect on growth performance but diarrhea incidences and severity were reduced (P > 0.05) in SDWE-fed piglets compared to the control. The results show that SDWE supported greater piglet performance pre-ETEC challenge although there was no benefit of SDWE or CPRO supplementation evident during the post-challenge period in early-weaned pigs. / October 2015

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第22074号 / 農博第2366号 / 新制||農||1072(附属図書館) / 学位論文||R1||N5228(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 近藤 直, 准教授 小川 雄一, 教授 飯田 訓久 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Made available in DSpace on 2014-08-20T13:32:55Z (GMT). No. of bitstreams: 1 tese_carine_dahl_corcini.pdf: 732454 bytes, checksum: 17c0395123a3b5438e40b167f5247599 (MD5) Previous issue date: 2010-06-25 / The knowledge of the reproductive potential of a male has economical and reproductive importance, especially if he is used in artificial insemination programs. The conventional assays for evaluating seminal quality do not have the capacity of measuring the fertilizing potential of a sample, they only indicate if it is within the acceptable parameters for insemination, according to the Brazilian College of Animal Reproduction. Into this context, the in vitro sperm penetration assay presents itself as an alternative laboratorial test to categorize fertile males regarding their fecundation capacity, since they mimic in vitro what happens in vivo. However, this assay has its use limited by the difficulty of execution and by the utilization of high cost equipment. The present work aimed to find a new biological model to study male reproductive aspects and to evaluate alternatives to simplify and to decrease costs of the assay execution: use of the chicken egg inner perivitelline layer (IPVL) penetration assay and its association with in vitro and in vivo parameters; use of BTS as incubation media for the in vitro penetration assay, and female gametes cryopreservation using different methods. It was concluded that: 1) Calomys laucha can be used as a biological model; 2) it is possible to use the inner perivitelline layer as a substrate in assays to predict fertility of Calomys laucha males; 3) the IPVL has receptors that allow swine sperm binding, therefore presents potential for use in male fertility diagnostic tests; 4) it is possible to use BTS as incubation media for the in vitro penetration assay, in oocytes or in IPVL, and in water-bath without CO2, and 5) the cryopreservation of ovaries at -20ºC or the refrigeration of oocytes in PBS at 5ºC before the execution of the in vitro oocyte penetration assay, using mTBM media, is an alternative to the use of fresh oocytes in tests to predict swine semen fertility. / O conhecimento do potencial reprodutivo de um macho é de importância econômica e reprodutiva, principalmente se ele é utilizado em programa de inseminação artificial. Os testes convencionais que avaliam a qualidade seminal não possuem a capacidade de medir o potencial fertilizante de uma amostra e, apenas indicam se a mesma se encontra dentro dos parâmetros aceitáveis para a inseminação segundo o Colégio Brasileiro de Reprodução Animal. O teste de penetração espermática in vitro, aparece neste contexto como um teste laboratorial alternativo para categorizar os machos férteis, quanto a sua capacidade de fecundação, pois mimetizam in vitro o que acontece in vivo. No entanto, este teste tem seu uso limitado por ser de difícil execução e por utilizar equipamentos de alto custo. O presente trabalho objetivou encontrar um novo modelo biológico para estudos sobre aspectos reprodutivos de machos e avaliar alternativas para simplificar e diminuir custos para execução do teste: utilização do teste de penetração na membrana perivitelina interna do ovo da galinha (IPVL) e sua associação com parâmetros in vitro e in vivo; utiilização do BTS como meio de incubação do teste de penetração in vitro e utilização de diferentes maneiras de criopreservação de gametas femininos. Foi concluído que: 1) Calomys laucha pode ser utilizado como modelo biológico; 2) é possível utilizar a membrana perivitelina interna como substrato de teste para predizer a fertilidade de machos Calomys laucha; 3) a IPVL possui receptores que permitiram a ligação do espermatozóide suíno, apresentando potencial para ser utilizada em diagnóstico de fertilidade de machos; 4) é viável utilizar BTS como meio para realização do teste de penetração in vitro em ovócito ou IPVL, em banho-maria sem a presença de CO2 e 5) o congelamento de ovários a -20ºC ou o acondicionamento a 5º C dos ovócitos em PBS para realizar o teste de penetração ovocitária in vitro, utilizando o meio mTBM, é uma alternativa para predizer a fertilidade de sêmen suíno.