Clostridioides difficile (C. difficile) and human rotavirus (HRV) are leading causes of bacterial and viral gastroenteritis worldwide. Treatment and vaccination options for both pathogens have significant limitations. C. difficile infections are treated with antibiotics, which is paradoxical as C. difficile itself is associated with antibiotic usage. In the United States, two live oral attenuated vaccines (Rotarix and RotaTeq) are licensed for protection against HRV. Since receiving approval from the World Health Organization (WHO), Rotarix and RotaTeq have been widely implemented into global national childhood immunization schedules, with one report finding 59 countries using Rotarix and 25 using RotaTeq. However, these vaccines have much lower efficacy rates in low- and middle-income countries. Because of these caveats, there is an urgent need to generate novel prophylaxes and treatments for C. difficile and HRV. In order to address this need, animal models that replicate the nuances of each infection are imperative. We have developed gnotobiotic (Gn) pig models for each pathogen. Gn pigs infected with spores of the hypervirulent UK1 strain of C. difficile develop classical signs of infection, including watery diarrhea and weight loss. Gross necropsy reveals colonic distention and discoloration, and histopathological evaluation shows volcano lesions, pseudo membrane formation, and epithelial cell erosion. Gn pigs infected with a G4P[6] strain of HRV also display pathogen specific signs of infection, including diarrhea, fecal rotavirus shedding, and damaged intestinal villi. A dose response study of the G4P[6] strain revealed diarrhea and virus shedding occurred at all tested doses, however the most severe diarrhea and virus shedding, measured by cumulative diarrhea score, area under the curve (AUC) of diarrhea, peak virus titer, and AUC of virus shedding, were all detected in the highest dose group. Based on the presentation of clinical signs of infection, 105 fluorescent focus units was selected as the optimal challenge dose for future studies. These models enable us to test candidate therapeutics, but also elucidate unique replicative features of the pathogens. For example, we found that HRV can replicate in the salivary glands and nasal cavity of Gn pigs in addition to the small intestine. HRV infection primed immune responses in the ileum, tonsils, and facial lymph nodes; infection also induced high levels of systemic and mucosal rotavirus specific antibody responses. Moving forward, we hope to expand upon this replication study to identify what cell types within the glands are infected as well as look at local cellular immune responses to HRV infection. Additional future directions include determining the protective efficacy of next generation HRV vaccines and evaluating effectiveness of an engineered probiotic yeast in reducing severity of C. difficile infection and disease. The Gn pig models of C. difficile and G4P[6] HRV are clinically relevant, and they will continue to serve as useful tools to better our understanding of pathogenesis, infection, and prevention of these pathogens. / Doctor of Philosophy / Clostridioides difficile (C. difficile) and human rotavirus (HRV) both cause gastrointestinal related symptoms when they infect humans. Treatments available for C. difficile and HRV all have significant drawbacks. This represents a gap in knowledge which we aimed to fill by developing germ-free (gnotobiotic [Gn]) pig models of C. difficile and HRV infection and disease. Animal models that mimic the outcomes of disease seen in humans are essential for evaluating protectiveness of new therapeutics. The more similar the disease presentation, the more likely the treatment results will be translational to humans. We began with C. difficile; pigs were orally fed C. difficile and monitored for a week post infection for development of signs of infection. Inoculated pigs lost weight and developed diarrhea. Bacterial cells and toxins were isolated from fecal samples collected on various days post infection. Multiple changes were observed in infected pigs’ large intestinal tissues, including severe bleeding, tissue distension, and fluid buildup. Infected pigs also had significant upregulation of pro-inflammatory cytokines, indicating activation of the immune response. We performed a similar procedure for the establishment of the HRV model. Gn pigs were orally challenged with differing doses of G4P[6] HRV and followed for several days post infection. Consistent with HRV infection in children, the pigs developed watery diarrhea that lasted for multiple days. Small intestinal tissues collected at necropsy had several signs of damage, including blunted villi, fluid buildup, and immune cell invasion. These lesions were also consistent with HRV infection in humans. Taken all together, these results indicated successful establishment of both C. difficile and HRV models. While the primary goal of generating these models was to evaluate new treatments, a secondary goal was to use them to better our understanding of pathogen replication dynamics. For example, the small intestine was thought to be the primary site of HRV infection. Using a pig model of HRV, we expanded on this knowledge to show that HRV can replicate in the nose and salivary glands as well. Additionally, we found HRV infection to induce immune responses near the sites of infection, including the intestine, the tonsils, and the facial lymph nodes. Overall, these studies demonstrate the utility of germ-free pig models and are an important first step in generating more effective treatments for bacterial and viral infections.

Thesis (PhD)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: The aim of this study was to explore proteomics and microbiome profiling in pigs fed high fibre diets. In the first phase, maize cobs were ensiled using whey, molasses and exogenous enzymes in the laboratory and effects on ensiling characteristics and fibre levels were evaluated. In the second phase South African Windsnyer-type Indigenous pigs (SAWIP) and Large White x Landrace crosses (LW x LR) were fed diets containing ensiled maize cobs and evaluated on; diet preferences, nutrient digestibility and colonic fermentation; growth performance, carcass traits and blood metabolite profiles; the faecal microbiome; and serum and liver proteomic profiles. Ensiling maize cobs with molasses, whey and exogenous enzymes did not improve fermentation characteristics but exogenous enzymes reduced fibre fractions and energy content of maize cob silages. Dets containing two levels of maize cobs ensiled without any additive; a low (LMC) and high (HMC) maize cob inclusion levels and a control diet which did not have any maize cobs (CON) were formulated. The SAWIP preferred the CON diet more than (P < 0.05) diets with maize cobs while the LW x LR had no feed preferences. There was no correlation between preference and diet digestibility in both breeds. The SAWIP digested nutrients better (P < 0.05) than the LW x LR in the high fibre diets. There were no differences in the diversity of the core composition of gut bacterial communities between the breeds and diets. There were differences in the ratios of Bacteroidia to Clostridia between the SAWIP and LW x LR. Verrucomicrobiae was present in SAWIP and LW x LR on HMC diet and not on the CON diet. There was a breed x diet interaction (P < 0.05) for Oscillospira. Analysis of the microbiome revealed breed differences and no dietary differences. There were differences in serum and liver proteins and in serum metabolite levels. Two specific proteins identified were Guanidinoacetate N-methyltransferase-like isoform 1 associated with creatine biosynthetic and Catalase, which is involved in cholesterol metabolic processes. At the grower stage, the SAWIP consumed more feed per metabolic body weight than the LW x LR while at the finisher stage LW x LR consumed more feed per metabolic body weight (P < 0.05) than the SAWIP. The breed of pig influenced most of the growth performance and carcass parameters more than the diet did. The SAWIP demonstrated an adaptation to high fibre diets by consuming more feed than the LW x LR per metabolic body weight at the grower stage. The inclusion of ensiled maize cobs in diets did not negatively affect selected commercial pork cuts. Analysis of faecal microbiomes revealed differences that may explain the enhanced ability of the SAWIP to digest fibrous diets better than the LW x LR breed.Proteomics can identify biomarkers that evaluate the performance of pigs consuming high fibre diets. A proof of principle to assess serum and liver protein profiles of pigs fed a a high fibre diet using a sodium dodecyl sulphate polyacrylamide gel electrophoresis matrix-assisted laser desorption ionization mass spectrometry (SDS-PAGE /MALDI MS) workflow was established. Key words: ensiling, exogenous enzymes, palatability, fermentation, fibre, metageome, biomarkers.

Thesis (PhD (Animal Sciences))--Stellenbosch University, 2011. / ENGLISH ABSTRACT: Local pigs in Southern Africa are an important component of resource-based subsistence farming systems and contribute substantially to the improvement of livelihoods of farmers. The objective of the study was to characterise indigenous pigs through the following specific objectives: to characterise the production systems, to give a physical description and to evaluate the genetic differentiation of the indigenous pigs. Surveys were carried out in Chirumhanzu and Mutoko Districts of Zimbabwe, Afred Nzo, OR Tambo and Vhembe districts of South Africa. Blood samples were collected in all of the above and additional three districts in Malawi (Mchinji, Dedza and Salima). The first study showed that most of the indigenous pigs were kept by women. The farmers kept small herd sizes (<7 pigs) to match the available resources. Income was the main determinant of farmer production objectives and breed preference. Several constraints that would militate against in situ conservation included poor quality and quantity of feeds, diseases, lack of housing, lack of markets and lack of support services. The pigs were generally small and black resembling the Windsnyer-Mukota type of pigs. The pigs apparently had a high foraging ability and high thermo-tolerance that made them suitable for production in low-intensity management free range production systems. These types of pigs were distributed throughout the study area. A microsatellite analysis showed high diversity but very little population differentiation among the pig populations from Southern Africa, with 93 % of variety occurring within subpopulations. Development of markets can be a feasible way of mainstreaming the indigenous pigs into the general economy. This will achieve the twin objectives of conserving and improving the breed while, at the same time, benefitting the farmers that keep these genetic resources. Farmers faced similar production constraints and the pigs were similar across the study areas. / AFRIKAANSE OPSOMMING: Plaaslike varke speel ‘n belangrike rol in hulpbron-gebaseerde bestaansboederye in Suider- Afrika, en maak ‘n aansienlike bydrae tot verbetering van die lewensbestaan van bestaansboere. Die studie het die volgende doelwitte: om die produksie van inheemse varke te karakteriseer, die varke fisies te beskryf, en om die genetiese differensiasie van die inheemse varke te evalueer. Opnames is uitgevoer in die Chirumhanzu en Mutoko distrikte van Zimbabwe, Afred Nzo, OR Tambo en Vhembe distrikte in Suid-Afrika. Bloedmonsters is in al die bogenoemde en nog drie distrikte in Malawi (Mchinji, Dedza en Salima) versamel. Die eerste studie het getoon dat die meeste inheemse varke deur vroue aangehou word. Die boere het klein trop groottes vir aanpasbaarheid by die beskikbare hulpbronne. Inkomste en varkras voorkeur was die hoof bepalende faktore vir hierdie boere se produksie doelwitte. Verskeie beperkings wat bots teen in situ instandhouding sluit in swak kwaliteit- en kwantiteit voere, siektes, gebrek aan behuising, die gebrek aan markte en die gebrek aan ondersteunende dienste. Die varke is oor die algemeen klein en swart en vertoon soos die Windsnyer-Mukoto tipe varke. Hierdie varke het blykbaar ‘n hoë voer-soekende vermoë en hitte-verdraagsaamheid wat hulle geskik maak vir die produksie in lae-intensiteit bestuur en vry-weidende produksie sisteme. Hierdie tipe varke was versprei oor die studie area. ‘n Mikrosatelliet analise het aangedui dat daar hoë genetiese variasie is binne die vark populasie, maar daar is klein differensiasie tussen die verskillende vark populasies van Suider-Afrika, met 93% variasie wat voorkom binne sub-populasies. Die ontwikkeling van markte kan ‘n haalbare manier wees om die inheemse varke toegang te gee tot die algemene ekonomie. Hiermee kan beide doelwitte, bewaring en verbetering van hierdie varkras, tergelyktydig bereik word, ten goede van die boere wat hierdie genetiese hulpbronne aanhou.

Master of Science / Department of Animal Sciences and Industry / Joel DeRouchey / Thirteen experiments using a total of 7,589 nursery and finishing pigs were conducted to evaluate the effects of deoxynivalenol (DON), feed additives and processing conditions on nursery pig growth performance. In addition, feed withdrawal and diet blending were evaluated in finishing pigs. Experiment 1 tested 3 feed additives in DON-contaminated diets with only Defusion Plus improving performance. Experiment 2 evaluated Biofix in both low- and high-DON diets and showed no effects on growth. Experiments 3 and 4 further evaluated levels of Defusion and the effects of pelleting and supplemental nutrients in DON-contaminated diets. Defusion improved growth in low-DON diets, but had variable effects in high DON diets. Pelleting DON-contaminated diets resulted in comparable growth to pigs fed positive control diets in meal form. In Exp. 5 and 6, pilot studies evaluated DON-detoxification using sodium metabisulfite (SMB) with hydrothermal treatment in both an autoclave and a pellet mill. These conditions reduced analyzed DON by as much as 89 and 75% for the autoclave and pellet mill, respectively. In Exp. 7 and 8, pelleting DON-contaminated diets with SMB improved growth. Experiments 9 and 10 evaluated feed-withdrawal time on carcass composition and economic returns. These experiments showed that pre-slaughter fasting for up to 36 h prior can be used to avoid weight discounts in heavyweight pigs without negatively impacting carcass composition and maintaining overall revenue. However, these advantages come with a potential reduction in carcass weight and increased incidence of leaking ingesta, which can result in condemned heads. Experiments 11, 12, and 13 compared phase-feeding to blending diets using an automated feed delivery system. These studies showed that corn-supplement blending is not economical and feeding diets blended to a Lys curve results in lower feed costs compared to phase-feeding, but due to reductions in growth and carcass weight, these savings do not translate into higher income over feed cost. Finally, Exp. 13 showed that over- and under-budgeting situations do not significantly influence overall returns, but pigs fed under-budgeted diets performed more closely to those fed correctly estimated feed budgets.

Schwanzbeißen ist eines der gravierendsten Probleme in der konventionellen Schweinehaltung, da es zu einer erheblichen Einschränkung des Tierwohls führt und zudem wirtschaftliche Einbußen nach sich zieht. Eine Prävention von Schwanzbeißen ist äußerst schwierig, da die Ursachen multifaktoriell sind. Viele Risikofaktoren, die Schwanzbeißen auslösen können, sind durch Untersuchungen hinreichend bekannt, aber das fehlende Bewusstsein für das Vorhandensein dieser Risikofaktoren auf den Betrieben erschwert es den Landwirten, eine Veränderung der Situation herbeizuführen. Ein ganzheitlicher Ansatz ist erforderlich, um die vorhandenen Risikofaktoren auf den Betrieben zu identifizieren und für jeden Betrieb ein entsprechendes individuelles Profil mit seinen Stärken und Schwächen zu erstellen. Vor diesem Hintergrund wurde das Schwanzbeiß Interventions Programm (SchwIP) 2011 entwickelt, dem eine betriebsindividuelle Erhebung verbunden mit den Grundsätzen der Planung von Tiergesundheit und Tierwohl (Animal Health and Welfare Planning, AHWP) zu Grunde liegt. SchwIP wurde auf deutschen konventionellen Betrieben mit Mastschweinehaltung angewendet und evaluiert, um Landwirten zu helfen, die Risikofaktoren für Schwanzbeißen zu identifizieren und zu reduzieren, sowie sich der Herausforderung zu stellen, die Gegebenheiten auf den Betrieben zu optimieren. Das Ziel der vorliegenden Arbeit war es zu ergründen: 1) wie eine Management-Anwendung in der Praxis verbreitet werden kann und ob ein Wissenstransfer zu den Anwendern erfolgt, 2) ob die Anwendung von SchwIP bezüglich einer Reduzierung von Risikofaktoren für Schwanzbeißen und einer Verringerung der Prävalenz von Schwanzverletzungen auf den Betrieben effektiv ist und 3) ob sich die Effektivität der Anwendung der Management-Hilfe auch in der Erfassung der Prävalenz von Schwanzverletzungen am Schlachthof widerspiegelt. Es wurden 23 eintägige Schulungen mit 19 Veterinären und 115 landwirtschaftlichen Betriebsberatern durchgeführt, um die Management-Hilfe SchwIP in ganz Deutschland zu verbreiten (Kapitel 4). Die Teilnehmer erlernten die Anwendung von SchwIP in interaktiven Schulungen mit einer Kombination aus theoretischen und praktischen Lehrabschnitten. Die Schulungsgruppen wurden subjektiv, basierend auf der Ausprägung ihrer ablehnenden Einstellung gegenüber Bestandteilen oder der Konzeption von SchwIP einer der folgenden drei Kategorien zugeordnet: hochgradig, mittelmäßig oder geringfügig ablehnend. In den nach der Schulung ausgeteilten Feedback-Bögen bewerteten hochgradig ablehnende Schulungsgruppen die Anwendbarkeit der Managementhilfe auf den Betrieben signifikant schlechter als die anderen beiden Gruppen. Allerdings wurden keine Unterschiede zwischen den Kategorien der Schulungsgruppen hinsichtlich der Benotung des Wissenstransfers in den Feedbackbögen sowie der sich an die Schulungen anschließenden praktischen Anwendungen auf den Betrieben festgestellt. Insgesamt erachteten 67% der Teilnehmer Schulungen als geeignetes Medium für das Erlernen zukünftiger Managementhilfen. Als Schlussfolgerung lässt sich ableiten, dass es erforderlich ist, sich Erfahrungen und Grundkenntnisse im Kommunikationstraining sowie in dem Leiten einer Gruppe anzueignen, bevor Schulungen entwickelt und durchgeführt werden, um auf die unterschiedlichen Einstellungen der Teilnehmenden eingehen zu können. Die Managementhilfe SchwIP wurde auf 188, von Schwanzbeißen betroffenen Betrieben, an jeweils einem Tag zwischen Juni und November im Jahr 2012 und erneut im gleichen Zeitraum im Jahr 2013 angewendet. Die Anwendung erfolgte entweder von einer Wissenschaftlerin der Arbeitsgruppe (68 Betriebe) oder von einem der 68 geschulten Tierärzte bzw. Betriebsberater (120 Betriebe) (Kapitel 5). Die Auswahl der untersuchten Buchten war problemorientiert, das heißt, Buchten wurde Vorrang in der Erhebung gegeben, in denen Schwanzbeißen zum Zeitpunkt des Besuchs vorhanden war oder Buchten, bei denen ein wiederkehrendes Auftreten bekannt war. Insgesamt zeigte sich, dass durch die Anwendung von SchwIP, bezogen auf alle Betriebe, das Gesamtrisiko zwischen den beiden Erhebungen signifikant reduziert werden konnte. Die Landwirte nahmen sich bei jedem Betriebsbesuch Ziele und Maßnahmen vor, unterstützt von dem jeweiligen Anwender und dem generierten SchwIP-Betriebsbericht, um das Risiko für Schwanzbeißen auf ihren Betrieben zu reduzieren. Der Grad der Umsetzung der vorgenommenen Maßnahmen (alle, einige oder keine Maßnahmen umgesetzt) hatte keinen signifikanten Einfluss auf die Veränderung des Gesamtrisikos von Schwanzbeißen. In den verschiedenen Risikokategorien Komfort, Beschäftigung, Futter & Wasser, Gesundheit und Stress konnte kein Unterschied im jeweiligen Gesamtrisiko der Kategorien zwischen den Erhebungen festgestellt werden. Allerdings konnte in der Kategorie Beschäftigung ein Wissenstransfer durch die Anwender und das SchwIP verzeichnet werden, da im Jahr 2013 mehr Betriebe eine Beschäftigung anboten, sowie mehr Betriebe Beschäftigungsmaterial (z. B. Stroh, Heu, etc.) anstelle von Beschäftigungsobjekten (z. B. Kette mit Holzstück) bereitstellten. Darüber hinaus zeigte sich, dass sich die Prävalenz von Schwanzläsionen (Blut am Schwanz, entzündliche Schwellungen des Schwanzes, Teil- oder Vollverlust) und von Ohrläsionen (Blut oder Kruste am Ohr) zwischen den beiden Betriebserhebungen signifikant reduzierte. Begleitend zu den Betriebserhebungen wurde eine Bonitierung der Schwanzläsionen von 32 Betrieben, auf denen das SchwIP angewendet wurde, und von 32 Kontrollbetrieben am Schlachthof durchgeführt (Kapitel 6). Drei Beobachter beurteilten Schwanzläsionen von einer Gesamtstichprobe von 80.034 geschlachteten Schweinen anhand von Fotos, wobei 43.402 Fotos von SchwIP Betrieben im Zeitraum vom 02.07.2012 bis 29.11.2013 bonitiert wurden und 36.632 Fotos von Kontrollbetrieben im Zeitraum vom 22.11.2012 bis 29.11.2013. Die Bonitur erfolgte mit Hilfe einer 4-stufigen Skala (keine / leichte / schwere Verletzung, Nekrose). Zusätzlich wurde beurteilt, ob ein Vollverlust des Schwanzes vorhanden war oder nicht. Für die Auswertung wurden die Verletzungsgrade zusammengefasst zu den Befunden ‘Verletzung‘ und ‘keine Verletzung‘. Verletzungen wurden unabhängig von dem jeweiligen Schweregrad im Durchschnitt bei 25,4 % der Schlachtkörper von Kontrollbetrieben festgestellt, wobei die Mehrzahl der Verletzungen leichte Verletzungen waren (23,6 % der Schlachtkörper). Es wurden weniger Schwanzspitzennekrosen in der routinemäßigen Fleischuntersuchung ermittelt als Nekrosen von Fotos bonitiert wurden. Von insgesamt 548 Schwanzspitzennekrosen wurden nur 17 % übereinstimmend sowohl von Fotos als auch in der routinemäßigen Fleischuntersuchung erfasst, wohingegen 53 % der Nekrosen, die in der routinemäßigen Fleischuntersuchung festgestellt wurden, auch in der Fotobonitur als solche bewertet wurden. Die Prävalenz von Schwanzverletzungen war in der ersten Saison (Winter), in der sowohl von SchwIP- als auch von Kontrollbetrieben Fotos vorhanden waren, im Beobachtungszeitraum signifikant höher bei den SchwIP-Betrieben als bei den Kontrollbetrieben, aber in keiner der folgenden Saisons. Dies weist auf eine Reduzierung durch die Anwendung der Managementhilfe SchwIP hin. Zusammenfassend erwies sich die Kombination von einer betriebsindividuellen Erhebung, entsprechend, den in SchwIP integrierten Grundsätzen der Planung von Tiergesundheit und Tierwohl (AHWP) als erfolgreich. Die Risikofaktoren für Schwanzbeißen auf Betrieben mit bestehender Schwanzbeißproblematik konnten reduziert und die Prävalenz von Schwanzverletzungen, sowohl auf den Betrieben als auch im Schlachthof, gesenkt werden. Schulungen mit Betriebsberatern und Veterinären erwiesen sich zudem als ein sehr effektiver Weg, neue Managementhilfen in der landwirtschaftlichen Praxis zu verbreiten und einen Wissenstransfer von der Wissenschaft in die Beratung und daran anschließend an die Landwirte bereitzustellen.

Šiuo tyrimo tikslas buvo ištirti laukinių ir naminių kiaulių halotano geną. Tyrimo metodika. Tyrimas atliktas 2012-2013 m. LSMU Biologinių sistemų ir genetinių tyrimų institute K. Janušausko gyvūnų genetikos laboratorijoje. Viso, HAL geno atžvilgiu, ištirta 137 kiaulė (45 šernai, 5 Vietnamietiškos kiaulės, 12 Pekariai, 25 Landrasai, 25 Jorkšyrai ir 25 Pjetrėnai). Kiaulių šeriai buvo naudojami kaip genetinės medžiagos šaltinis. Halotano geno nustatymui buvo atliekama polomerazės grandininės reakcija (PGR). HAL geno įtakai mėsinėms ir penėjimosi savybėms įvertinti buvo atrinkti mišrūnai LxJxP (45) auginti Valstybinėje kiaulių veislininkystės stotyje. Penėjimosi ir mėsinių savybių duomenys gauti iš Valstybinės kiaulių kontrolinio penėjimo stoties Kauno skyriaus. Rezultatai ir išvados. Iš 75 tirtų naminių kiaulių 88 proc. buvo NN ir 12 proc. Nn genotipo. Visos (62) laukinė kiaulės buvo NN genotipo. Aukščiausias n alelio dažnis nustatytas pas Pjetrėnų (p<0,01) veislės kiaules lyginant su Landrasais ir Joršyrais. Tuo tarpų N alelio didžiausias dažnis rastas pas Jorkšyrų veislės kiaules. Kadangi visos tirtos laukinės kiaulės neturėjo streso geno (n) alelio galima daryti prielaidą, jog jos yra atsparesnės stresui. Mūsų tyrimų duomenimis HAL geno homozigotinių ir heterozigotinių kiaulių mėsinių ir penėjimosi savybių rodikliai skiriasi. Šiltos skerdėnos masė, skerdenos puselės ilgis ir bekono puselės ilgis buvo didesni pas NN genotipo namines kiaules. Pas heterozygotinius individus... [toliau žr. visą tekstą] / The aim of present study was to research halothane gene in wild and domestic pigs. Research methodology. Research was performed during 2012 – 2013 year in LSMU Biological Systems and Genetic Research Institute of K. Janusauskas Animals Genetic Laboratory. Samples for DNA testing were collected from wild and domestic pigs. Fattening and meat quality data were obtained from the State of Pigs Control fattening Station of Kaunas Department. A total of 137 pigs (45 wild boars, 5 Vietnamese pigs, 12 Pecari, 25 Landrace, 25 Yorkshire and 25 Pietrian) were tested for halothane genotype. For the effect of HAL gene on meat and fattening properties, 45 crossbreeds (LxJxP) from State Pigs Breeding Station were selected. Plucked hair was used as a source of genomic DNA. All tested animals were characterized as normal homozygotes (NN), heterozygotes (Nn) or recessive homozygotes following amplification of a target region of the HAL gene using the polymerase chain reaction (PCR), followed by a restriction endonuclease assay. The resulting PCR was digested with the restriction enzyme Alw21I, followed by agarose gel electrophoresis. Results and conclusions. In 75 tested domestic pigs, 88 % were NN and 12 % were Nn genotype. In 62 tested wild pigs all animals were NN genotipe. The frequency of n allele was higher (p<0.01) in Petrian pigs (0.25 for n) than in Landrase (0.1 for n) and Yorkshire (0.05 for n). The frequency of N allele was higher in Yorkshire than in the rest of the tested... [to full text]

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This diploma thesis deals with the importance of tourism for a group of states which includes Portugal, Spain, Italy and Greece. The aim of this thesis is to identify the importance of tourism to the economy and to interconnect it with the competitiveness of those countries in tourism via an analysis of the current situation in tourism and via analysing the effects that tourism has on these economies in the past last five years. The first chapter explains the most important terms concerning the subject in theory. The next chapter introduces tourism within the PIGS economies and the third chapter is devoted to tourism competitiveness of the southern EU countries and creates a predicition of their future development in tourism.

This study occurred during July, August, and September in the Small Animals Investigation and Production Center (CIPAM), dependent of the Faculty of Agricultural, Livestock, and Veterinary Sciences of the Technical University of Oruro, of the Central Highland of Bolivia. In order to satisfy guinea pig nutrition requirements, which are 2800.00 kcal/kg of energy, 18.00% protein, 10.00% fiber, 1.00% calcium, and 0.60% phosphorus, the following ingredients were used in feed: 11.30% yellow corn, 30.00% milled wheat [Bromus catharticus], 14.00% soy cake, 32.00% wheat bran, 9.00% alfalfa flour, 1.80% bone flour, 1.00% conchilla, 0.50% common salt, and 0.40% methionine. The effect that vitamin C deficiency causes is scurvy, with 100% mortality of guinea pigs at 28 days without green forage. This investigation was made with supplementation of canahua, wheat, and soybean sprouts in the growth stage with the object of replacing green forage in winter by presenting vitamin C. Canihua sprouts at 8 days gave a maximum of 6.28mg of vitamin C /100g of sample, and wheat sprouts at 8 days gave a maximum of 16.09mg of vitamin C/100g of sample. Soybean sprouts at 13 days reached 21.41mg of vitamin C /100g of sample, which continued to increase, but one should be warned that maintaining sprouts of this species is expensive. Rations consisted of 90.00g of sprouts plus 72.00g of feed per guinea pig, with the oral administration of 12.98mg of vitamin C in water solution per guinea pig used in the area. The reason is the improvement in guinea pig meat production with high protein content (20.30%), compared to cattle (17.50%) and sheep (14.50%), and the lower cholesterol of guinea pig (7.80%) compared to cattle (21.80%) and sheep (37.80%), and at a lower cost, with organic sprouts of high nutritional value. The canahua-soybean sprout treatment with 45% dry matter, 5.73% ash, 7.03% ether extract, 25.50% protein, 6.85% raw fiber, and 51.58% free nitrogen extract at 64 days of winter growth showed a live weight gain of 437.73g in relation to pure feed of 66.05% dry matter, 7.33% ash, 2.83% ether abstract, 19.51% raw protein, 7.52% raw fiber, and 56.46% free nitrogen extract with 359.23g. The difference is 78.50g, which is significant at 0.05 with a t-test. The result indicates that the canahua-soybean interaction is the best for live weight gain at 64 days of growth with respect to normal feed, for which the null hypothesis is rejected and the alternative hypothesis accepted. The live weight gain by sex in guinea pigs at 48 and 64 days of growth was highly significant, in females from 206.67g (4.3g/day) to 303.29g (4.74g/day) and in males from 262.27g (5.46g/day) to 392.27g (6.13g/day), with differences between males and females of 55.6 and 89g. These results show that male guinea pigs generally gain more weight than females during growth, which could be commercialized in the opportune moment. The food conversion between sexes fed with the different sprouts at 64 days of growth was 9.55 in females and 7.62 in males, a difference of 1.90. Also, the sex-soybean interaction in females with soybean was 10.31 and in males with soybean 6.79, a difference of 3.52, which is significant at 0.01. Similarly, the food conversion at 64 days of treatment by canahua sprouts and a feed with 41.34% dry matter, 5.69% ash, 3.47% ether extract, 20.58% raw protein, 8.04% raw fiber, and 56.25% free nitrogen extract is 7.90. In relation to just the feed it is 9.20, with a difference of 1.30. In a t-test this is significant at 0.05. These results show that male guinea pigs eat less food and grow more than females under the same conditions. The lowest production cost was obtained with canahua-feed interaction, in females Bs4.0/guinea pig (US$0.501) and in males Bs3.9/guinea pig (US$0.488). Canahua-soybean interaction yielded Bs5.50/guinea pig (US$0.689) for females and Bs5.50/guinea pig (US$0.689) for males. Normal feed yielded Bs4.20/guinea pig (US$0.526) for females and Bs5.40/guinea pig (US$0.676) for males. (US$1 = Bs7.98). Also, there was a percentage variation for the canahua-soybean-feed interaction in females of 5.00% lower than just feed and in males 27.80% lower than just feed. These results show that it is best to use canahua sprouts plus feed because this gives the lowest costs of feeding growing guinea pigs in winter.

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