Thesis (MSc. Agriculture (Animal Production)) -- University of Limpopo, 2015 / Three experiments were conducted to determine the effect of in ovo glucose injection on egg hatchability, chick hatch-weight, productivity and carcass characteristics of indigenous Potchefstroom koekoek chickens. A complete randomized design was used in all the three parts of the study (from incubation, 1-49 days old unsexed chickens and 50-91 days old female chickens). On day 18 of incubation the developing eggs were subjected to the following treatments: 0- (no glucose or water injected), 0+ (only water injected), 5, 10, 15 or 20 mg of glucose per egg. Each treatment had three replications and there were 20 eggs per replicate. A quadratic model was used to determine in ovo glucose injection levels for optimal egg hatchability, chick hatch-weight and chick to egg weight ratio of Potchefstroom koekoek chickens. In ovo glucose injection improved (P<0.05) egg hatchability, chick hatch-weight and chick to egg weight ratio of the chickens. Egg hatchability, chick hatch-weight and chick to egg weight ratio Potchefstroom koekoek chickens were optimized at different injection levels of 4.50, 10.43 and 12.00 mg of glucose per egg, respectively. Unsexed day-old chicks from the first experiment (according to their initial treatments and replicates) were used in a complete randomized design having six treatments, replicated three times, and having ten birds per replicate. Glucose injection levels increased (P<0.05) feed intake, growth rate, feed conversion ratio, live weight, metabolisable energy and nitrogen retention of female Potchefstroom koekoek chickens aged 1 to 49 days. However, growth rate, live weight, metabolisable energy intake and nitrogen retention of the chickens were optimized at glucose injection levels of 3.92, 4.36, 10.67 and 13.50 mg per egg, respectively. Female chickens from the second part of the study (according to their initial treatments and replicates) were used in a complete randomized design having six treatments, replicated three times, and having five birds per replicate. In ovo glucose injection levels improved (P<0.05) on feed intake, growth rate, feed conversion ratio, live weight, metabolisable energy and nitrogen retention of female Potchefstroom koekoek chickens aged 50 to 91 days. However, only feed conversion ratio and metabolisable energy intake of the chickens were optimized at glucose injection levels of 12.15 and 5.57 mg per egg, respectively. Injection level also improved (P<0.05) carcass, breast, v drumstick, thigh, wing, gizzard and liver weights of female Potchefstroom koekoek chickens aged 13 weeks. In ovo glucose injection increased (P<0.05) breast meat tenderness, juiciness and flavour of female Potchefstroom koekoek chickens aged 91 days. However, breast tenderness, juiciness and flavour of female Potchefstroom koekoek chickens were optimized injection levels of 13.50, 19.25 and 10.83 mg of glucose per egg, respectively. No chicken deaths were observe

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21967号 / 農博第2357号 / 新制||農||1070(附属図書館) / 学位論文||R1||N5218(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 清水 浩 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

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Introduction: The desire to have a biological child transcends race, religion and socio-economic status. However for those faced with infertility, the financial resources needed to conceive are often not available. Current research in assisted reproduction has gravitated towards cost reduction to address restricting financial factors, without compromising quality of treatment. One such initiative is the development of a low-cost embryo culture method by The Walking Egg foundation. This method utilizes a standard chemical reaction and simple equipment to equilibrate culture media pH and to regulate temperature; both aspects were investigated in this study. An exploration into the insemination concentration to achieve oocyte fertilization was also undertaken. Methods: Quality control of temperature regulation on six different heating devices, including a comparison of inter- and intra-variations was carried out. The utilization of citric acid and bicarbonate of soda for carbon dioxide production, which subsequently facilitate setting of pH values, was tested by injecting increasing citric acid volumes (1.2 ml – 3.0 ml in 0.2 ml increments) into set volumes of bicarbonate of soda. Further investigation evaluated gas production at various temperatures (37°C, 25°C and 15°C), at increasing intervals (16 – 30 hours) of equilibration and these were compared by measuring pH of the culture media. The influence of altitude on pH was explored by repeating the chemical reaction experiment at five different locations in South Africa. Furthermore, the addition of water to citric acid before gas generation was explored. The minimal insemination concentration needed for fertilization was determined by the addition of decreasing numbers of spermatozoa to non-fertilized bisected oocytes. The experiment was repeated with a selected sperm insemination number in 1 ml or 50 μl culture media to compare the tested culture system with conventional culture. Spermatozoa bound to the hemi-zonae were counted with the aid of an inverted phase contrast microscope. Hemi-zonae with bound sperm were also stained with ethidium homodimer and evaluated using a confocal laser-scanning microscopy system. After removal of hemi-zonae, the spermatozoa in culture were isolated for deoxyribonucleic acid fragmentation analyses and reactive oxygen species presence in the culture media was measured. Additionally, reactive oxygen species generation in simulated culture was measured over time. Results: All the equipment tested bar one, the warming oven, proved useable with the simplified Walking Egg in vitro fertilization culture system. By decreasing the citric acid volumes, it was indicated that 1.8 ml citric acid, diluted with 1.2 ml water, is the optimal volume to facilitate the required culture media pH. Omitting the water dilution from citric acid volumes affected the culture media pH adversely, however reducing the temperature during gas equilibration did not. A change in altitude had no effect on culture media pH. Lower insemination numbers resulted in decreased sperm binding, with 2 x 103 motile sperm insemination providing the lowest number to still obtain sufficient sperm–zona binding (≥20 sperm bound). Incubation in 1 ml vs. 200 μl culture media indicated decrease in sperm bound. Sperm deoxyribonucleic acid fragmentation and the presence of reactive oxygen species in the culture media were similar in both the test and control groups. A comparison over time revealed less reactive oxygen species in 1 ml culture media, from the simplified Walking Egg in vitro fertilization culture system after three days of culture, than 200 μl culture media drops under oil, from conventional culture after 18 hours, however the results were not statistically significant. Discussion: Purpose-made heating devices provide superior stabilization of culture media temperature. When selecting a heating device, intra-variations should be considered. Culture media can be manipulated to the required pH by carbon dioxide production, with meticulous attention paid to the citric acid volumes used. However, if gas generation is performed at room temperature, equilibration time must be increased. In conventional culture, the minimum insemination number can be reduced to 2 x 103 motile sperm. Due to lower binding of sperm in large volumes of culture media, 2 – 5 x 103 motile sperm should be considered for the simplified culture system, depending on a holistic consideration of all sperm parameters. Extended culture for at least three days with the simplified culture system can be performed without increasing reactive oxygen species present in culture media. Further research of this novel culture method should include the application of the culture method in a South African environment. / Dissertation (MSc)--University of Pretoria, 2017. / Obstetrics and Gynaecology / MSc / Unrestricted

MSCAGR (Animal Science) / Department of Animal Science / In order to realize many of the potential advantages of AI, storage of semen is necessary. Semen storage is only possible using a system that decreases and/or halts the metabolic processes of the spermatozoa, allowing no significant loss of fertility. Numerous factors affect the success of spermatozoa storage. This study was designed to compare the effects of egg yolk, soybean milk and coconut water in Tris extender using different storage methods for Nguni bull spermatozoa storage. Bull semen was collected from two adult Nguni bulls approximately four years old and kept under similar managerial conditions. Using electro-ejaculator, semen was collected from each bull into a graduated semen collection tube. Macroscopically evaluation of the sample was performed immediately after collection. Only the semen free from contamination was processed. The kinetic properties namely: total spermatozoa motility, and progressive spermatozoa motility were analysed using CASA. Semen sample was stained and spermatozoa morphology and vitality also analysed using CASA. The extended semen was then split into three groups. The first group was stored at room temperature (25 °C). The second group was cooled to 4 °C and stored in the refrigerator. The third group was also cooled to 4 °C for 2 h in the refrigerator, then held in LN2 vapour 5 cm above the surface of LN2 at ~ -80 °C for 10 min and then plunged into LN2 for storage at -196 °C. Different colours of straws and plugging powder were used for identifying each extender. After 3 days of storage at room temperature, in the refrigerator and in LN2, the extended semen was split into three portions and assayed for kinetic properties using the first portion. The second portion was assayed for spermatozoa morphology and the third portion for spermatozoa vitality. The results from the fresh semen extended with all three extenders (TEYE, SBME and COWE), and analysed immediately after dilution at room temperature (25 ºC), showed no significant difference (P > 0.05) in the mean values of the kinetic and morphologic properties and viability, on spermatozoa TM, PM, AR, AT, CT; BT and LS. After three days of storage, there was no significant difference (P > 0.05) in the kinetic morphologic properties and viability of semen stored at room and refrigeration temperature regardless of the extender in use. There were, however, significant differences (P < 0.05) in the TM, PM, AR and DL of the frozen semen samples. For the short storage period of semen used for AI, from this study, it is recommended that semen should be kept at room or refrigeration temperature regardless of the three extenders used. However, for long storage of frozen semen TEYE is recommended. The egg yolk-based extender provided greater preservation of motility and bull spermatozoa integrity during the freezing process than did SBME and COWE.

Both during the ovogenesis and immediately after the laying is a bird egg exposed to strong pressure from the microorganisms which are able to penetrate the egg through eggshell pores and infect its inner structures. With regard to the proved negative effects on hatchability, viability and phenotype of offspring, a cascade of obstacles has evolved which are able to minimize the risk of bacterial infection. The deposition of antimicrobial components into the eggshell and the egg white is considered to be one of them. A whole group of egg white proteins belong to them, among which lysozyme and ovotransferrin, which are also partially deposited into the eggshell and cuticular layer of the eggshell, dominate with their antimicrobial effect. The implication of the newest studies is that the microbial protection of the egg can be also performed by the eggshell pigments - biliverdin and protoporphyrin, which protect the egg from being colonized by microorganisms with an increased UV light and regulation of conductivity and water vapour condensation on the egg surface. Even though the presence of these antimicrobial components in different structures of the egg has been proved, there is no known comparative study which addresses the relation between deposition of selected antimicrobial components into...

In addition to proving an excellent source of nutrients, eggs are used in the food, cosmetic, and biotechnology industries for their rheological and bioactive properties. Much of the potential for the added value is in individual components of the egg, rather than the whole egg. At low speed centrifugation, yolk separates into two distinct fractions—granules and plasma. It is becoming increasingly popular in the industry to remove the plasma fraction of the egg yolk to use for its livetins, particularly immunoglobulin Y, leaving behind a granule by-product (“yellow cake”). Previous research has shown potential added-value from the granule fraction, especially from its phosvitin and phospholipids. Granules are protein aggregates with complexes of phosvitin and high density lipoproteins linked by phosphocalcic bridges. In their native form, the proteins are mostly insoluble, however previous studies have shown the links can be broken by alterations in pH, ionic strength, and mechanical treatments. This thesis project seeks to find potential uses for the egg yolk by product after the removal of the livetin fraction by means of further fractionation with mechanical treatment (filtration). Two variables were tested to extract more proteins from the yellow cake. Salt was added to 10% solids solution of yellow cake in water before filtration at four different NaCl levels: 0%, .05%, 1%, and 2.5%. Additionally pH was tested at four different levels: 4.6, 4.8, 5.0, 5.2. The samples were also tested for antibacterial properties against Escherichia v coli with a minimum inhibitory concentration assay (MIC). Analysis with BCA showed salt concentration had a significant effect on the yield of protein. The highest concentration of salt tested, 2.5%, had the highest protein yield. Additionally, SDS PAGE showed 2.5% salt had the most unique protein bands. This could be to the disruption of the phosphocalcic links between the phosvitin and HDL by NaCl, allowing the protein to solubilize. pH did not have a significant effect on the yield or types of proteins in the range tested in this experiment. There is no conclusive evidence of antibacterial properties against E. coli from the protein extract. The MIC assay had growth show up in all wells with the protein extract, however there was a visible decrease in turbidity with higher concentration of the protein extract. This could mean that the protein extract does have some antibacterial properties, but needs testing at higher concentrations or with isolated proteins/peptides. The SDS-PAGE revealed bands showing phosvitin present, which has known antibacterial properties. Overall, improvements to the methods for further protein extraction from egg yolk by-products will help lead the industry to finding novel uses and product applications.

An experiment was conducted to determine whether direct-fed microbial supplementation could alleviate a marginal amino acid (AA) deficiency in Hyline 36 laying hens from 33 to 44 wk of age. The experiment was a 2 × 4 factorial design with or without a commercial directfed microbial (Primilac; 1.36 kg/1,000 kg) and 4 levels of AA formulation. Egg characteristics (yolk, albumen, or shell proportions and yolk or albumen solids) were not affected by diet. Primilac supplementation had no effect on egg production or egg mass. However, Primilac supplementation reduced feed intake-to-egg mass ratio by 2.4 and 3.4% from 33 to 36 wk and 41 to 44 wk, respectively. Total eggs laid and egg mass were greatest when at least 14.4 g of CP, 804 mg of Lys, 382 mg of Met, 601 mg of TSAA, 502 mg of Thr, and 609 mg of Ile were consumed per hen per day from 33 to 44 wk of age. In conclusion, Primilac supplementation was not able to completely alleviate a marginal AA deficiency in laying hens but did improve feed intake-to-egg mass ratios during 8 wk of the 12-wk study.

A control diet (CN diet; 17.4% CP and 4.37% Ca) was compared with an experimental diet with a 0.4-percentage-unit reduction in protein content and a 35% replacement of limestone by a CaSO4-zeolite mixture [5.76% CaSO4 and 1.18% zeolite; reduced-emission diet (RE diet)] in laying hens to evaluate the effects on apparent N retention, egg production, egg composition, and excreta pH measured at excreta collection or after 7 d of storage. In previous studies, it was demonstrated that the RE diet reduced NH3 emissions by 48%. Laying hens (192 total; 48 replicate cages per diet, with 2 hens per cage) were fed experimental diets from 33 to 49 wk of age. Apparent N retention (48.2%), egg production (83.6%), and number of shell-less eggs (0.18%) were not affected by the diet. Eggs tended to be heavier (59.4 vs. 58.8 g/egg, P = 0.06), and yolk percentage (29.7 vs. 29.0%, P = 0.013) was greater with the RE diet. At 48 wk of age, the total solids content per egg was also greater from hens fed the RE diet (13.2 vs. 12.6 g/egg, P = 0.032). Other egg components were not influenced by diet. Thus, a slight reduction in dietary CP content and replacing a portion of the Ca from CaCO3 with CaSO4 did not affect egg production nor did it impair shell quality. At the end of the experiment, excreta were collected from all cages (excreta from 3 cages were mixed and pooled; 16 pools of excreta per diet). At collection, excreta of hens fed the RE diet had lower pH (5.89 vs. 6.54, P < 0.001) than those of hens fed the CN diet. After 7 d of storage, excreta pH of hens fed the RE diet continued to be lower (6.30 vs. 8.36, P < 0.001). The reduction of excreta pH, even after 7 d of storage, may control nitrogenous emissions from excreta by maintaining excreted N as NH4+.

Nutrifish es un proyecto que nace de la idea de brindar nutrición a los consumidores, entre ellos, su principal público objetivo son las madres amas de casa encargadas de llenar las despensas del hogar. A lo largo del camino, encontramos dificultades, pros y contras de realizar la comercialización del producto, sin embargo, con las herramientas y conceptos básicos aprendidos a lo largo de la carrera, pudimos determinar que el balance era positivo y la rentabilidad atractiva. Nutrifish fue el resultado de realizar una lluvia de ideas, las cuales abarcaban desde temas de salud, hasta opciones lúdicas, por ende, los cimientos del proyecto eran bastante débiles, se fueron fortaleciendo a medida que tomábamos conocimiento sobre el mercado de productos precocinados y el manejo interno de sus marcas. A continuación, les mostraremos el camino recorrido por nuestro grupo de trabajo. / Nutrifish is a proyect born from the idea of taking nutrition to a certain number of costumers, its main target are the housewifes, in charge of filling the house storerooms. Along the way we’ve found some difficulties, pros and cons of the commercialization of the product, however, the tools and basic concepts learnt to this day, we found ourselves able to establish a positive balance and an attractive profitability. Nutrifish was the result of a brainstorming, the group went from healthy themes to playful options, thus, the foundations of the project were certainly weak, and we strengthened them by taking knowledge of the precooked food market and the internal management of their brands. In the following paragraphs, we’ll show you the way taken by our work group. / Trabajo de investigación