Molting in crustaceans is a highly complex physiological process involving negative regulation by two paired endocrine glands, the X-organ/sinus gland complex (XO/SG) and the Y-organ (YO). The XO/SG complex is responsible for making molt-inhibiting hormone (MIH) which negatively regulates synthesis of the molting hormones, ecdysteroids, by the YO. Analysis of gene expression in the XOs and YOs has led to the development of a proposed molecular signaling pathway which regulates ecdysteroidogenesis and subsequent molting in crustaceans. In this study, changes in protein abundance in the YO were characterized over the course of a molt cycle (intermolt, early premolt, mid premolt, and late premolt) induced by multiple leg autotomy (MLA) in the blackback land crab, Gecarcinus lateralis. In all, 457 distinct protein spots were detected in the molting gland using two-dimensional gel electrophoresis, of which 230 (50%) changed significantly in abundance over the course of the molt cycle (one-way permutation ANOVA, p≤0.05). Changes in protein abundance were most notable between the intermolt and the three premolt stages, indicative of a biological ‘on-off’ switch in the Y-organ. Several hemolymph species proteins, including hemocyanin, cryptocyanin, and transglutaminase, were identified which characterized physiological changes associated with molting beyond the Y-organ. An abundance of cytoskeletal proteins were identified which correspond with glandular hypertrophy and are indicative of vesicular-mediated exocytosis, possibly of ecdysteroids. Further, several proteins involved in the immune, proteostasis, and oxidative stress response are characteristic of supporting the dynamic and demanding cellular changes associated with ecdysteroidogenesis and the transition of the Y-organ from the basal to the highly active state. Many proteins involved in energetic pathways including glycolysis, the citric acid cycle, amino acid metabolism, and one-carbon metabolism changed in abundance in response to both the higher energy demands and the requirement for precursors of macromolecular synthesis of the YO over the molt cycle. Taken together, these changes in diverse physiological pathways represent the complexity involved with regulation of the Y-organ, even with just the single proposed physiological purpose of ecdysteroidogenesis.

Diese Arbeit befasst sich mit der Entwicklung einer Hochdurchsatz-Methode für Herstellung und Screening imprägnierter heterogener Katalysatoren. Es werden Hopkalit-, Platin- und Chrom-basierende Stoffsysteme auf Aluminiumoxid bzw. Titandioxid untersucht. Heterogene Katalysatorbibliotheken mit diskreten Zusammensetzungsgradienten werden mittels Tintenstrahldruck präpariert, wofür zunächst bedruckbare poröse Substrate hergestellt werden. Es wird eine Methode zur teilautomatisierten Bestimmung der Größe dosierter Mikrotropfen implementiert. Gedruckte Katalysatoren werden mit klassisch trockenimprägnierten Proben verglichen. Für eine homogene Verteilung der Katalysatorspezies werden Einflussparameter evaluiert, von welchen die Tintenviskosität als entscheidend identifiziert wird. Zur Einschätzung der Katalysatorhomogenität wird Raman-Mikrospektroskopie eingesetzt. Für Untersuchungen unter Reaktionsbedingungen wird ein Parallelreaktor konstruiert und gefertigt. Zum Vergleich dienen Versuche im klassischen Strömungsrohr. Als Testreaktion dient die Kohlenmonoxid-Oxidation. Die Aktivität der imprägnierten Katalysatoren wird mittels In-situ-IR-Spektroskopie mit verschiedenen Messstrategien untersucht. Versuche an Katalysatorpulvern im konventionellen Strömungsrohrreaktor ermöglichen einen Vergleich der Ergebnisse.:1 Einleitung 2 Theoretischer Teil 2.1 Tintenstrahldruck 2.1.1 Funktionsweise von Tintenstrahldruckern 2.1.2 Druckbarkeit von Fluiden mittels DOD-Tintenstrahldruck 2.1.3 Benetzungsverhalten gedruckter Tropfen auf Feststoffoberflächen 2.2 Kaffeeringeffekt 2.2.1 Ursache des Kaffeeringeffekts 2.2.2 Unterdrückung des Kaffeeringeffekts 2.3 Hochdurchsatz und Kombinatorik 2.3.1 Tintenstrahldruck in der kombinatorischen Materialforschung 2.3.2 Hochdurchsatz-Screening heterogener Gasphasenreaktionen 3 Ergebnisse und Diskussion 3.1 Konzept und Zielsetzung 3.1.1 Konzept des Drucks binärer Katalysatorbibliotheken 3.1.2 Vorangegangene Arbeiten 3.1.3 Übersicht und Zielsetzung dieser Arbeit 3.2 Vorversuche an klassisch imprägnierten Katalysatoren 3.2.1 Charakterisierung der eingesetzten Substratmaterialien 3.2.2 Thermogravimetrische Analyse der Katalysatorimprägnierung 3.2.3 Raman-Spektren ausgewählter imprägnierter Katalysatoren 3.2.4 Pulverröntgendiffraktogramme ausgewählter Katalysatoren 3.3 Substratherstellung 3.3.1 Herstellung von Dünnschichtsubstraten 3.3.2 Herstellung von eingestrichenen Substraten 3.3.3 Porenvolumina der Substrate 3.4 Tintenstrahldruck heterogener Katalysatorbibliotheken 3.4.1 Bestimmung der Mikrotropfengröße 3.4.2 Allgemeine Vorgehensweise zur Bestimmung der Katalysatorverteilung 3.4.3 Einfluss der optischen Fokussierung auf die Raman-Intensität 3.4.4 Drucken heterogener Katalysatoren auf Dünnschichtsubstraten 3.4.5 Drucken heterogener Katalysatoren auf Presslingen 3.4.6 Drucken heterogener Katalysatoren auf eingestrichenen Substraten 3.5 Kohlenmonoxid-Oxidation und Mikrospektroskopie 3.5.1 Aufbau der Reaktionszelle 3.5.2 In-situ-Infrarot-Spektroskopie 3.5.3 In-situ-Raman-Spektroskopie 4 Zusammenfassung und Ausblick 5 Experimenteller Teil 5.1 Tintenstrahldruck von Katalysatorbibliotheken 5.1.1 Allgemeines zum eingesetzten Drucksystem 5.1.2 Substratherstellung 5.1.3 Vorbereitung der Vorstufenlösungen 5.1.4 Einstellung der Dosierparameter und Koordinatenkalibrierung 5.1.5 Ermittlung der Tropfengröße 5.1.6 Erstellung des Druckprogrammes und Durchführung des Druckprozesses 5.2 Verwendete Analysemethoden 5.2.1 Raman-Mikrospektroskopie 5.2.2 Infrarot-Mikrospektroskopie 5.2.3 Porenvolumenbestimmung mittels Helium-Pyknometrie 5.2.4 Stickstoffadsorption zur Bestimmung von Feststoffoberfläche und Porenvolumen 5.2.5 Partikelgrößenbestimmung 5.2.6 Pulverröntgendiffraktometrie 5.2.7 Thermogravimetrische Analyse 5.3 Bestimmung der katalytischen Aktivität 5.3.1 Reaktionszelle für In-situ-Spektroskopie 5.3.2 Umsatzbestimmung von Pulverproben im konventionellen Strömungsrohr 5.4 Verwendete Chemikalien 6 Anhang Verzeichnisse Literaturverzeichnis Abkürzungsverzeichnis Abbildungsverzeichnis Tabellenverzeichnis Selbständigkeitserklärung Danksagung

Limited breeding success in captive breeding programs has necessitated the development of assisted reproductive technologies (ART) to preserve and increase genetic variation and population numbers of both captive and wild amphibian groups. ART has been shown to be successful in numerous anuran species, and current studies focus on the application of ART in ex-situ populations. The focus of this project is to show that linking in-situ and ex-situ amphibian populations through sperm cryopreservation, genome banking, and in-vitro fertilization is possible, with the goal of increasing gene diversity throughout groups in order to produce self-sustaining, wild populations in the future. Specific objectives include developing a sperm-cryopreservation methodology using sperm from the model species Anaxyrus fowleri, applying this protocol to the cryopreservation of spermatozoa from two other threatened anurans to determine protocol transmissibility, and linking in-situ and ex-situ populations of an endangered species using cryopreserved sperm form wild males to produce viable offspring.

The prevalence of non-alcoholic fatty liver disease (NAFLD) in children has increased over the past decades, creating a need for animal models that recapitulate the features of the pediatric disease. Iberian pigs have a leptin-resistant phenotype characterized by hyperleptinemia, hyperphagia, and extreme adipogenesis. We hypothesized that neonatal Iberian pigs fed a high fat high-fructose (HFF) diet will develop a pattern of liver injury resembling pediatric NAFLD. In addition, we sought to determine if a mixture of probiotics would prevent the disease. Animals were fed 1 of 4 diets containing (g/kg body weight × d) 0 g fructose, 11 g fat and 199 kcal (CON-N; n=8), 22 g fructose, 16 g fat and 300 kcal (HFF2-N; n=8), CON + probiotic (CON-P; n=6), or HFF2 + probiotic (HFF2-P; n=6) every 6 h for 70 d. The probiotic mixture (6.2 × 104 cfu/mL) contained Pediococcus acidilactici, Pediococcus pentosaceus, Lactobacillus plantarum and Bacillus amyloliquefaciens. Body weight was recorded every 3 d. Serum markers of liver injury and dyslipidemia were measured on d 40 and 65 at 2 h post feeding. Fasting leptin, insulin, glucose and homeostatic model assessment (HOMA) values were assessed on d 70. Liver and skeletal muscle (longissimus dorsi) were collected on d 70 for histology, triacylglyceride (TAG) quantification, relative gene expression, and Western blot analysis. Metabolomic analysis was performed on liver tissue and plasma. Body weight was not significantly greater in HFF fed pigs compared to CON. Leptin, alanine and aspartate aminotransferases, alkaline phosphatase, lactate dehydrogenase and total bilirubin were increased (P ≤ 0.001), and high and low density lipoproteins decreased (P ≤ 0.05) in HFF2-N and HFF2-P. Livers in HFF2-P and HFF2-N had higher relative weight and TAG (P ≤ 0.001), micro and macrovesicular steatosis, ballooning degeneration, Mallory-denk bodies, inflammation and necrosis, increased gene expression of TNFα, TGFβ, IL1α and PPARγ (P ≤ 0.001), and decreased ChREBP (P ≤ 0.001). A probiotic affect was seen as pigs fed CON-P and HFF2-P had higher insulin and HOMA values were increased (P ≤ 0.05). Western blot analysis showed dysregulation of autophagy in liver of pigs fed CON-P and HFF2-P, and in skeletal muscle of pigs fed CON-N and HFF2-N. Metabolomic analysis demonstrated dysregulation of one-carbon metabolism, the tricarboxylic acid cycle (TCA), the urea cycle, and amino acid metabolism of pigs fed HFF2 diets compared to CON diets. In conclusion, Iberian pigs fed a HFF diet recapitulate many pediatric NAFLD-associated features, in the absence of obesity and independently of probiotic supplementation, suggesting a potentially suitable model for pediatric NAFLD research. Furthermore, probiotic supplementation did not ameliorate the onset of NAFLD when fed in conjunction with a HFF diet.

Thesis (M.Sc. (Animal Production)) -- University of Limpopo, 2023 / Two experiments were conducted to determine the effect of citric acid inclusion level in the diet on growth performance, carcass weight, tibia bone breaking strength and ash content of male Ross 308 broiler chickens aged one to 35 days. The first experiment determined the effect of citric acid inclusion level in the diet on growth performance traits of male Ross 308 broiler chickens aged one to 21 days. The experiment commenced with 200 male day-old Ross 308 broiler chickens with an initial average live weight of 40±1.6g per chick. The chicks were assigned to five treatment groups in a completely randomized design, each replicated five times, and each replicate having ten chicks. The citric acid inclusion levels were at 0, 12.5, 25 or 50g per kg DM of feed. The second experiment determined the effect of citric acid inclusion level in the diet on growth performance, carcass weight, tibia bone breaking strength and ash content traits of male Ross 308 broiler chickens aged 22 to 35 days. The experiment commenced with 180 male Ross 308 broiler chickens aged 22 days. The chickens were assigned to four treatment groups, each having three replicate pens of eight chickens per replicate in a completely randomized design. Data was analysed using the General Linear model (GLM) procedures of the Statistical Analysis of System, version 9.3.1 software program. Where there were significant differences (P<0.05) between the treatment means, Tukey Multiple Comparison Test was used for mean separation. Citric acid inclusion in the starter diets improved (P<0.05) live weight and growth rate of male Ross 308 broiler chickens aged one to 21 days. Citric acid inclusion in the starter diets did not affect (P>0.05) daily feed intake, body weight gain and feed conversion ratio of male Ross 308 broiler chickens aged one to 21 days. The inclusion of citric acid did affect (P<0.05) live weight, body weight gain, feed conversion ratio and growth rate of chickens aged 22 to 35 days. Citric acid inclusion levels used in the present study influenced (P<0.05) DM and CP digestibility, ME intake and N-retention of male broiler chickens aged 22 to 35 days. The results of the current study showed that citric acid inclusion in a diet improved (P<0.05) chicken bone morphology. Thus, positive relationships were observed between citric acid inclusion level and right tibia bone weight, diameter, calcium, phosphorous and Magnesium contents of chicken bones aged 35 days. There were positive relationships between citric acid inclusion level and breast weight of male Ross 308 broiler chickens aged 35 days. Further studies are recommended to ascertain these findings. / National Research Foundation (NRF)

Amidst the amphibian extinction crisis, in situ and ex situ amphibian species conservation initiatives utilize assisted reproductive technologies for optimal genetic management of captive and wild populations. Development of effective, simple, low-cost methods for obtaining gametes for artificial fertilization, sperm biobanking, or natural breeding alleviates obstacles for institutions or programs to apply reproductive technologies. Objectives herein include investigating the efficacy of alternative hormones for inducing spermiation, an alternative hormone administration route for inducing ovulation, exploring the physiological effects of hormone therapy, and utilizing sperm from deceased animals for biobanking.

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[pt] O ensino remoto estabelecido emergencialmente em função da pandemia de Covid-19 trouxe grandes desafios a educadores, que precisaram adequar as rotinas de ensino rapidamente para conduzir as aulas. No ensino em design, a experimentação material realizada em ambientes de laboratório de design é primordial para formação de estudantes de design de produtos que, por intermédio das construções físicas, exploram criativamente suas ideias ou as colocam em testes para a validação de conceitos e soluções de design, em um ambiente de ensino compartilhado por alunos, professores e profissionais técnicos. Esta dissertação apresenta o relato da experiência da autora na elaboração e implementação de alternativas de trabalho que permitiram a continuidade das práticas de experimentação material, por meio de representações físicas, em disciplinas amparadas pelo Laboratório de Modelos e Protótipos, pertencente ao curso de graduação em design de produtos, da PUC-Rio, no decorrer do isolamento social. O referencial teórico que sustenta a dissertação inclui autores como Donald Shön, Nigel Cross e Antônio Santoni Rugiu e por meio de metodologia participativa, utilizando pesquisa bibliográfica e documental, busca-se observar e refletir o lugar da experimentação em laboratórios de design e sobre os ensino aprendizados ocorridos nestes espaços, destacando-se a importância das representações físicas no desenvolvimento de projeto de produtos. Por fim, apresenta-se as propostas que foram implementadas em um contexto de ensino remoto emergencial, como forma de possibilitar que a rotina de trabalho dos alunos, em suas residências. / [en] The remote teaching established on an emergency basis due to the Covid-19 pandemic brought great challenges to educators, who needed to adapt teaching routines quickly to conduct classes. In design education, material experimentation carried out in design laboratory environments is essential for training students to design products that, through physical constructions, creatively explore their ideas or put them to tests for the validation of design concepts and solutions, in a teaching environment shared by students, teachers and technical professionals. This dissertation presents the report of the author s experience in the elaboration and implementation of work alternatives that allowed the continuity of the practices of material experimentation, through physical representations, in disciplines supported by the Laboratory of Models and Prototypes, belonging to the graduation course in design of products, from PUC-Rio, during social isolation. The theoretical framework that supports the dissertation includes authors such as Donald Shön, Nigel Cross and Antônio Santoni Rugiu and through participatory methodology, using bibliographical and documentary research, we seek to observe and reflect on the place of experimentation in design laboratories and on teaching -learning that took place in these spaces, highlighting the importance of physical representations in the development of product design. Finally, the proposals that were implemented in an emergency remote teaching context are presented, as a way to enable the students work routine, in their homes.