Currently, there is little known about stink bug (Hemiptera: Pentatomidae) eggs, their natural enemies, and their susceptibility to insecticides. A survey of stink bug egg parasitoids was conducted in row crops and vegetables in eastern Virginia. Parasitization was highest in Euschistus servus (Say) with 89.7% and 49.2% of egg masses and individual eggs parasitized, respectively, followed by Acrosternum hilare (Say), with nearly half of all individual eggs parasitized. The most common parasitoid was Telenomus podisi Ashmead (Hymenoptera: Scelionidae). Laboratory egg-dip bioassays and field applications of acephate, lambda-cyhalothrin, spinosad, and thiamethoxam, were carried out to determine efficacy against nonparasitized E. servus and A. hilare eggs, and T. podisi embryos developing in E. servus eggs. Results showed that eggs of both species were susceptible to insecticides, that there was little difference among insecticides, but there was generally greater mortality in field-treated versus dipped eggs. Developing T. podisi were generally more susceptible to insecticides than stink bugs. Scanning electron microscopy was used to investigate oviposition sites as possible sites of insecticide movement into eggs. Oviposition wounds and holes made by a tungsten probe were similarly sealed by a "scab", so it was not clear whether these wounds allow for increased insecticide movement into parasitized eggs. Differences in chorion permeability of non-parasitized and parasitized eggs were compared by immersing them in solutions containing different ¹⁴C-ammended insecticides at field application rates for 0, 30, 120 or 240 minutes. Results showed that insecticide movement into the egg increased significantly with immersion time for both acephate and λ-cyhalothrin, but there were no significant differences between nonparasitized and parasitized eggs. Neither immersion time nor egg status was significant for thiamethoxam. A model was constructed that predicts amount of insecticide entering the egg at any given time. An 8-week survey for the brown marmorated stink bug, Halyomorpha halys (Stål) was conducted in Beijing and five other cities in China. Incidence of egg parasitism was recorded. Results showed that H. halys utilized at least four different plants throughout the summer, and insects were found in Nanjing, Kunming, and Xi'an. Parasitization of eggs was noted, and the parasitoids were identified as Trissolcus halyomorphae Yang (Scelionidae: Hymenoptera) by K.A. Hoelmer (USDA-ARS). / Ph. D.

Direct-fed microbials (DFM) are a method to increase bird performance and health when antibiotics are removed or limited in the diet. The objective of this experiment was to determine the effects of a single strain Bacillus subtilis DFM on pullet performance, layer performance, and egg quality when fed from day of hatch until 70 weeks of age. In total, 576 Hy-Line W-36 laying hen pullets were randomly divided between two treatments (Control and DFM) on day of hatch across 12 cages for each of the treatments and maintained on those respective treatments for 70-weeks. The DFM was added at a target dose of 300,000 cfu/kg of feed. At the end of the 17-week pullet phase, DFM fed pullets resulted in reduced feed intake, improved FCRm with both similar body weights (P = 0.98) and body composition (P > 0.05) to the control fed pullets. At 17 weeks, 72 birds were randomly selected from each treatment and moved to 12 laying hen cages. At the end of the 52-week laying hen phase, DFM-fed hens had significantly higher hen-day egg production (P = 0.02), but decreased egg weights. No differences were reported in egg mass or feed intake, but FCR was significantly improved (P = 0.02) on a per dozen eggs basis, and improved (P = 0.06) on a g/g basis by 7.1 points with DFM-fed hens having an FCR of 2.016 and control fed hens having and FCR of 2.087. Supplementation of diets with DFM resulted in a trend in egg specific gravity, but no differences were reported in shell breaking strength or shell weight. Overall, supplementation of the DFM lowered feed intake and improved FCR without affecting body composition in the pullet phase, and increased production and improved FCR during the egg laying phase. / Master of Science / Due to the decreased use of antibiotics across the poultry industry, producers and allied companies are developing new methods to increase bird health and performance when antibiotics are removed from or limited in the diet. One such method is through the use of "good" bacteria or technically called Direct-Fed Microbials (DFM). Direct-fed microbials are defined as live microorganisms that when fed, confer a health benefit to the host. The goals of this thesis were to determine the effects of a Bacillus subtilis based DFM on pullet performance, laying hen performance, and egg quality when fed from day of hatch until 70 weeks of age. In the pullet phase, DFM-fed pullets had reduced feed intake and improved feed conversion ratio (the ratio of feed intake to body weight gain), with no differences in body weights or body composition at the end of the 17-week growth period. In the laying phase, DFM fed hens had significantly improved egg production, but lighter eggs compared to the control fed hens. Addition of the DFM significantly improved feed conversion ratio per dozen eggs by 5.3 points and reduced feed conversion ratio on a g/g basis by 7.1 points in the 52-week laying phase. Small improvements were noted in specific gravity, but no differences were reported in shell weight or shell breaking strength. When fed from day of hatch until 70 weeks of age, the Bacillus subtilis DFM allowed the pullets to reduce their feed intake, but maintain body development for egg production resulting in an improved feed conversion ratio (FCR) in the growth phase before egg production and once hens initiated egg production resulted in smaller eggs, but increased egg production and FCR in laying hens with minimal differences in egg quality. Therefore, the DFM may be able to be used in place of growth promoting antibiotics to help support efficient protein production.

My dissertation and research focused on the effects of insecticides on squash bugs, its egg parasitoid, and pollinators in the production of cucurbits in Virginia. Plants in the cucumber family are dependent on insect pollination for successful fruit set, and are also susceptible to plant eating insects. Squash bugs are capable of transmitting cucurbit yellow vine decline, and their feeding can cause significant wilt and death in many varieties. To control for squash bug and other pests, growers commonly combine the application of broad-spectrum insecticides with the frequently applied prophylactic fungicides. Broad-spectrum insecticide applications are known to have negative effects on natural enemy populations, are capable of promoting insecticide resistance, and can have negative effects on pollinators if care in their use is not taken. Squash bugs have several natural enemies, but their predominant egg parasitoid is most effective at reducing damaging populations. The scelionid wasp Gryon pennsylvanicum Ashmead, is a prevalent egg parasitoid in Virginia and can be negatively affected by the application of broad-spectrum insecticides. Through survey efforts I found that G. pennsylvanicum is widely distributed throughout Virginia and is capable of high rates of egg parasitism (>90%). This is contrary to the 20% level previously assumed for the East Coast. I explored the effects of narrow-spectrum insecticides on the fate of the egg parasitoids, those developing in the host egg and emerged adults of G. pennsylvanicum. Contact assays showed that the insecticides λ-cyhalothrin and sulfoxaflor had caused high adult parasitoid mortality. As new insecticides get registered for use there is often concern about their effect on pollinators, specifically the European honey bee Apis mellifera L. I evaluated the use of large flight cages as a method to measure the sub-lethal effects of narrow-spectrum insecticides to honey bees, as a means to qualify risk. The method utilizes small colonies of honey bees (with stores of nectar and pollen) and their feeding at a treated sucrose solution after being trained to a feeder in an enclosed arena. This choice-test style behavioral experiment shows promise in qualifying the risks associated with insecticide exposure in the field. In the case of pyrifluquinazon, colonies repeatedly choose to avoid feeding at tainted feeders even after training with no other outside sources of food present. Further researching the sub-lethal behavioral effects that insecticides have on bees in a colony can help us better qualify their risk. / Ph. D.

The effect of dietary fats on reproductive performance, egg quality, fatty acid composition of tissues and egg yolk, and prostaglandin levels in embryonic tissues in Japanese quail (Coturnix, coturnix japonica) were evaluated. The addition of 5.0% chicken fat (CHX), hydrogenated soybean oil (HSBO), menhaden fish oil (MENH) or soybean oil (SBO) to the maternal and paternal diet altered tissue and yolk composition of the hens and tissue composition of the males to reflect the dietary source. Comparisons were made to chicken fat which served as a control. Feeding MENH increased the omega three fatty acid concentrations in liver, heart, testicle, and yolk. Feeding HSBO and SBO increased the omega six fatty acid concentrations in yolk. Feeding MENH and SBO increased the total monosaturated fatty acids in yolk. The concentrations of polyunsaturated fatty acids were increased by feeding MENH and SBO. Feeding MENH decreased egg production. Feeding MENH and HSBO decreased hatchability. Feeding MENH decreased specific gravity of eggs at day 30, 60, and 90. Feeding CHX increased specific gravity at day 30, 60 and 120. In addition, feeding CHX increased chick weight. Feeding SBO decreased early embryonic death. There were no consistent differences noted in tissue prostaglandin levels of embryos from hens on the differing diets. / Master of Science

Le jaune d’œuf est une source de nutriments et de molécules bioactives pour l’embryon. Il renferme de nombreuses protéases et antiprotéases, dont la fonction reste méconnue. Notre objectif était d’identifier et de caractériser les protéases/antiprotéases associées spécifiquement à la formation du jaune d’œuf (vitellogénèse). Les précurseurs du jaune sont exprimés par le foie à la maturité sexuelle des poules. Par une approche transcriptomique, nous avons identifié 582 gènes, dont 15 protéases et antiprotéases, sur exprimés par le foie des poules sexuellement matures. Au moins trois d’entre elles sont présentes dans le jaune ou la membrane vitelline. Nous avons montré que l’expression de la « similar to nothepsin », une protéase prédite de fonction inconnue, était spécifique du foie des femelles et pourrait participer à la maturation des précurseurs du jaune. D’autre part, nous avons démontré que l’ovoinhibiteur, une antiprotéase du jaune, aurait un rôle dans la défense antimicrobienne de l’œuf. L’intégration de l’ensemble de ces résultats fournira des données essentielles relatives à la vitellogénèse et à la fonction des protéases/antiprotéases dans le jaune d’œuf. / The egg yolk is a source of nutrients and bioactive molecules for the embryo,including many proteases/antiproteases, the function of which is still unknown. Our objective was to identify and characterize proteases/ antiproteases that are specifically associated with the formation of the yolk (vitellogenesis). Yolk precursors are synthesized by the liver at sexual maturity of hens. Using a transcriptomic approach, we identified 582 genes, including15 proteases and antiproteases that are over-expressed by the liver at sexual maturity of hens. At least three of them are recovered in the egg yolk or the vitelline membrane. We have shown that the expression of "similar to nothepsin” was specific to the liver of females. This protease could participate in the processing of yolk precursors. We have also demonstrated that egg yolk ovoinhibitor is antimicrobial and could therefore participate in egg defense. The integration of all these results will provide major data relative to vitellogenesis and to the function of proteases/antiproteases in the egg yolk.

Thesis (M.Sc. (Animal production)) --University of Limpopo, 2011 / A study was conducted to examine the effect of protein level of the diet of Naked neck hens on egg production, hatchability and chick productivity. The first part of the experiment determined the effect of protein level on feed intake, number of eggs produced, egg weight, egg white nitrogen content, egg yolk nitrogen content, egg hatchability and chick hatch-weight. A total of 25 hens aged 30 weeks were confined in cages and fed diets differing in protein content. A complete randomized design, comprising of diets differing in protein levels (P12.94%, P13.94%, P14.38%, P15.75% and P18.13% CP), replicated five times with one hen in each replicate was used. Semen was collected from one cock to fertilize all hens. Protein level had an effect on (P<0.05) feed intake and egg white nitrogen content. However, protein level of the diet of hens had no effect(P>0.05) on the number of eggs produced, egg white nitrogen content, egg yolk,egg weight, egg hatchability and chick hatch-weight. Quadratic analyses indicated that feed intake, number of eggs produced, egg weight, egg white nitrogen content, egg yolk nitrogen content, hatchability and chick hatch-weight were optimized at different protein levels of 14.7 (r2 = 0. 623),14.9 (r2 = 0.568),13.9 (r2 = 0.094), 18.2 (r2 = 0.563), 15.1(r2 = 0.424), 15.9 (r2 = 0.451) and 15.9 %(r2 = 0.898) , respectively. The second part of the experiment determined the effect of protein level of hens on productivity of their progenies from a day old up to 13 weeks of age. A total of 95 chicks hatched from the first part of the study were assigned to 20 floor pens according to the number of chicks hatched. Chicks had an average initial live weight of 35 ± 2 g per bird. The chicks were fed the same commercial grower diet and fresh water ad libitum up to seven weeks of age. Protein level of the diet of Naked neck hens had an effect (P<0.05) on feed intake, live weight and feed conversion ratio of chicks from a day old up to seven weeks of age. However, protein level of the diet of Naked neck hens had no effect (P>0.05) on growth rate and mortality of chickens from a day old to seven weeks of age. Feed intake, feed conversion ratio, growth rate and mortality of chickens were optimized atdifferent protein levels of 15.8 (r2 = 0.298), 16.1 (r2 = 0.236), 16.1 (r2 = 0.077) and 14.3 % (r2 = 0.617), respectively. Protein level of the diet of Naked neck hens had no effect (P>0.05) on feed intake, growth rate and feed conversion ratio of both their male and female progenies between eight and 13 weeks of age. Protein levels of the diet of hens had an effect (P<0.05) live weight of their male progenies and carcass weights of both their male and female progenies at 13 weeks of age. However, the level of protein of the diet of hens had no effect (P>0.05) on live weight of their male progenies, and breast meat yield, breast meat nitrogen content and fat pad weight of both their male and female progenies at 13 weeks of age. Live weight,breast meat yield, breast meat nitrogen content and fat pad weight of their male progenies at 13 weeks of age were optimized at different protein levels of the diets of hens of 18.9 (r2 = 0.666), 15.6 (r2 = 0.081), 15.4 (r2 = 0.786) and 17.7 %(r2 = 0.775), respectively. Similarly, live weight, carcass weight, breast meat yield, breast meat nitrogen content and fat pad weight of their female progenies at 13 weeks of age were optimized at different protein levels of 15.7 (r2 = 0.294),15.4 (r2 = 0.180), 15.8 (r2 = 0.059), 15.1 (r2 = 0.882) and 16.1 % (r2 = 0.405), respectively. It is concluded that protein level of the diet of the hen may affect its productivity and that of its progeny. However, the calculated protein levels for optimum productivity will depend on the production parameter in question. / National Research Foundation

Cryopreservation of germplasm is widely used in agriculture, biotechnology, conservation of threatened species and human reproductive medicine. There is a need however to improve the reproductive efficiency of breeding with cryopreserved semen, which may involve increasing the post-thaw quality of sperm through improvements in cryopreservation extenders. Extenders including egg yolk from chickens are successfully used worldwide for cryopreservation of bovine semen, whereas the protective agent in the egg yolk is believed to be the low-density lipoprotein (LDL) fraction. Egg yolks of different avian species vary in their cholesterol, phospholipid and polyunsaturated fatty acid content which have been shown to have important effects on sperm’s freezing capability. The purpose of this study was to determine the cryoprotective effect of clarified egg yolk and LDLs extracted from different egg yolk sources (chicken, chicken omega-3, pigeon, quail and turkey) on bovine sperm. Semen from six bulls was collected four times each by electroejaculation, split and diluted with the 10 following extenders: chicken clarified (Ccl), chicken omega-3 clarified (O3cl), pigeon clarified (Pcl), quail clarified (Qcl), turkey clarified (Tcl), chicken LDL (CLDL), chicken omega-3 LDL (O3LDL), pigeon LDL (PLDL), quail LDL (QLDL) and turkey LDL (TLDL). The extended semen was evaluated, cryopreserved and examined directly after thawing (0h) and after two hours at 37 ˚C (2h). Computer assisted sperm analysis (CASA) was used to determine total sperm motility (TM), progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL) and average path velocity (VAP). Intact plasma membrane (IPM) and intact acrosomes (IA) were measured by flow cytometry. The percentage change (loss; Δ%) of each sperm characteristic was calculated and used to compare the effect of the extenders. From extending to 0h post-thaw, the pigeon LDL extender lead to greater losses in sperm total and progressive motility, as well as of intact acrosomes, than the other nine extenders tested (P < 0.05). During 0h to 2h post-thaw, the sperm in PLDL extender experienced greater losses in total and progressive motility (P < 0.0001), as well as in curvilinear velocity (P < 0.05), than in all the other nine extenders. Sperm in turkey clarified extender had a greater loss in the velocity parameters (VSL, VAP, VCL) than sperm in several of the other extenders such as O3cl, CLDL, O3LDL, QLDL and TLDL from 0h to 2h (P < 0.05). Concomitantly, sperm in the Tcl extender had a greater loss in the velocity parameters and of intact acrosomes compared to sperm in its counterpart, the turkey LDL extender, from 0h to 2h post-thaw (P < 0.05). The differences produced in post-thaw quality of cryopreserved bovine sperm in the pigeon LDL and turkey clarified extenders were attributed to methodological differences in these egg yolk preparations compared with the other eight extenders. Importantly, the results demonstrate that with most egg yolk preparations derived from a variety of species, there are equivalent cryoprotective effects afforded by the use of omega-3 chicken, pigeon, quail, or conventional chicken egg yolk in a clarified form in freezing extenders for bovine semen. We further proved that the freezing capabilities of bovine semen extenders containing the low-density lipoprotein fraction of omega-3 chicken, quail, turkey and conventional chicken egg yolk were similar.

The reproductive success for pelagic spawning Baltic flounders is strongly linked to the hydrodynamics in the spawning areas. Egg survival is dependent upon the ability to achieve neutral buoyancy at a depth interval where temperature and oxygen concentrations are favourable for egg development. The main focus of this thesis was to compare egg survival probabilities of pelagic eggs from Baltic flounder in the Bornholm, Gdansk and Gotland basins, prior to and after the saline water inflow in December 2014. The results showed greatly enhanced survival probabilities in Bornholm basin 2015 (p&lt;0.01), as egg survival increased from 47% in 2014 to 100% the following year. In Gdansk basin the situation was similar, and survival probability increased from 13% to 100% (p&lt;0.01). In Gotland basin no difference in survival probability was identified, although the dominant cause of mortality shifted from sedimentation, i.e. due to low salinity conditions in 2014, to oxygen deficiency in 2015 (p&lt;0,01). / BONUS INSPIRE-project, the joint Baltic Sea research and development programme (Art 185), funded jointly by the European Union’s Seventh Programme for research, technological development and demonstration and the Swedish Research Council Formas

京都大学 / 0048 / 新制・課程博士 / 博士(情報学) / 甲第20025号 / 情博第620号 / 新制||情||108(附属図書館) / 33121 / 京都大学大学院情報学研究科社会情報学専攻 / (主査)教授 守屋 和幸, 教授 松田 哲也, 教授 廣岡 博之 / 学位規則第4条第1項該当 / Doctor of Informatics / Kyoto University / DGAM

In the modern poultry industry, intense genetic selection for meat production has negatively influenced the reproductive performance of commercial birds. Parthenogenesis, embryonic development in unfertilized eggs without any sperm-egg interactions, is known to hinder the normal fertilization process and could be one of the reasons for this reduced reproductive performance in the poultry industry. Therefore, the overall objective of this research was to gain a better understanding of the process of parthenogenesis using Chinese painted quail as the model. Studies on Chinese painted quail reproduction revealed that they are very inefficient in sustained sperm storage and that number of sperm penetrating the egg and subsequent embryonic development potentially alter egg transit time through the oviduct. This poor sperm storage capacity and high sperm-egg interaction requirement might be responsible for the occurrence of parthenogenesis in this species; and in fact, this makes Chinese painted quail an excellent choice for parthenogenesis research. Further, dams selected for parthenogenesis as well as embryonic development, including parthenogen size, alter egg components by possibly delaying the transit time of the egg through the oviduct. Also, both dams and sires selected for the parthenogenesis trait appear to influence their progenies performance, including 1st wk mortality and occurrence of parthenogenesis. Additionally, vaccination of virgin hens with live pigeon pox virus increases parthenogenesis as well as parthenogen size and livability by the direct action of the virus on the embryo. Moreover, live Newcastle disease virus under in vitro conditions was found to have similar effects on the embryo. Because parthenogenesis exists in the modern poultry industry, even the accidental selection of the trait in either males or females could have a negative impact on overall chick production and performance. Also, as vaccination is a routine practice in the industry, it is possible that vaccination of birds that carry the trait will reduce fertility and hatchability due to enhanced parthenogenesis. Overall, currently it appears that, parthenogenesis is adversely affecting the poultry industry; and therefore, additional research on the accurate determination of losses in the poultry industry due to parthenogenesis could further benefit the industry.