Frozen-thawed ram semen crosses the cervix poorly, necessitating laparoscopic insemination. Acceptable fertility can be achieved with frozenthawed ram semen deposited at the external cervical opening if ram seminal plasma is added. Homologous seminal plasma improves the fertility of frozen-thawed sperm of boars and dogs. Heterologous seminal plasma may have effects as well; the addition of bovine seminal plasma increases the ability of buffalo sperm (Syncerus caffer) to fertilize bovine oocytes in vitro. The aim of the current study was to compare the effects of seminal plasma of rams and bulls, dog prostatic fluid, protein-free TALP, TrilEq (Triladyl with 0.5 ml of Equex STM paste added to each 100 ml) and skim milk upon longevity and percentages of progressively and aberrantly motile frozenthawed ram sperm. Three ejaculates from each of 6 rams (2 Dorpers, 2 Döhne merinos, and 2 merinos), aged 2 to 4 years, were extended in TrilEq, pooled and frozen as a single batch per ram at 200 × 106/ml in 0.25 ml straws. Seminal plasma of rams was obtained from the same rams, while seminal plasma of five bulls were obtained by centrifugation of their ejaculates and dog prostatic fluid consisted of the post-sperm fractions of the ejaculates of 5 dogs. Within a 10 species, the seminal plasma or prostatic fluid from different donors was pooled and frozen in aliquots at −18 °C. The 108 straws (6 rams, 6 diluents, 3 replicates) were thawed in random order. Once thawed, a straw was emptied into a tube with 0.85 ml of the appropriate medium at 37 °C and kept at that temperature for 6 h. The percentage of progressively motile sperm was estimated at ×200 magnification immediately (time zero) and 2, 4 and 6 h after thawing. One person thawed the semen and prepared motility specimens, while another performed all motility evaluations. Data were evaluated by means of repeated-measures ANOVA, with rams as subjects and time and medium as fixed effects. Non-significant interactions were removed from the model. Pairwise comparison of means was done by means of Bonferroni's test (P < 0.05). The model included Ram, Time, Medium, and Ram × Medium, and Time × Medium interactions, which were all significant (P < 0.01). Mean progressive motility decreased from each time to the next and were 39.0% (0 h), 26.0% (2 h), 19.6% (4 h) and 12.6% (6 h); SEM 1.38%, n = 108. Mean motility was higher for skim milk (39.9%) than for all other media except TrilEq (27.7%), which was better than bull seminal plasma (13.0%), whereas TALP (20.5%) and ram seminal plasma (21.9%) were similar to TrilEq and bull seminal plasma (SEM 2.85%, n = 72). The interactions (Ram × Medium or Time × Medium) were mainly due to dog prostatic fluid, ram seminal plasma, TrilEq, and TALP, while milk resulted in the best and bull seminal plasma in the lowest motility. This study shows that heat-treated skim milk maintains progressive motility of frozen-thawed ram sperm better than dog prostatic fluid and seminal plasma of bulls and rams, TrilEq and protein-free TALP. In contrast to ram seminal plasma, skim milk is known to result in poor fertility of frozenthawed ram semen after cervical insemination. It would thus appear that maintenance of progressive motility in vitro may be a poor indicator of fertility after cervical insemination. / Dissertation (MSc (Veterinary Science))--University of Pretoria, 2006. / Production Animal Studies / MSc / unrestricted

En alimentation humaine, le poisson représente non seulement une source importante de protéines mais il apporte aussi des acides gras essentiels et des minéraux. Actuellement, en France, il est majoritairement consommé sous forme fraîche et préparé en filets sans peau. Cependant, face à la diminution des captures, à l'augmentation de la demande et à l'évolution des modes de consommation, l'importation de produits de la mer est de plus en plus importante dans notre pays. Or, depuis quelques années, les importateurs suspectent des fraudes consistant à vendre des filets de poisson décongelés sous la dénomination "frais". Ces produits entrent ainsi en concurrence directe avec les produits de la pêche française. L'objectif de la thèse consiste à mettre au point et à optimiser des méthodes de différenciation des filets de poisson frais de filets décongelés. La technique de l'électrophorèse bidimensionnelle comparative couplée à la spectrométrie de masse nous a permis d'identifier la parvalbumine comme marqueur de différenciation frais/décongelé à partir des exsudats de filets de bar (Dicentrarchus labrax). Nous avons utilisé la composition des exsudats comme source potentielle d'autres indicateurs pour différencier les filets frais des filets décongelés. Nous nous sommes ainsi intéressés à différents paramètres tels que l'activité de l'α-glucosidase lysosomique (marqueur historique), le dosage du calcium libre et le dosage des nucléotides et de leurs dérivés, des protéines et des parvalbumines. Nous avons procédé à une analyse statistique par Classification Hiérarchique Ascendante (CHA) et nous avons ainsi mis en évidence trois groupes dissimilaires : les indicateurs de lyse cellulaire, les indicateurs d'altération des nucléotides et les indicateurs d'altération des protéines. Nous disposons ainsi d'outils de différenciation frais/décongelé complémentaires, rapides et peu onéreux susceptibles de répondre aux attentes des industriels de la filière. / Inhuman diet, seafood is an important source of proteins, essential fatty acids and minerals. Nowadays, in France, fresh fish is mainly consumed as skinless fillets. Due to the decrease of the fishing and the increase and evolution of fish consumption, the importation of fish becomes more significant in our country. Since several years, the importers suspect fraudulent pratices consisting in selling thawed fish fillets labeled as fresh ones. These products are directly in competition with the national fish market. The main aim of this thesis consisted in developing and improving methods to differentiate fresh versus frozen/thawed fish fillets. A comparative two-dimensional electrophoresis and tandem mass spectrometry proteins identification strategy, performed on fish fillet exudates of European sea bass (Dicentrarchus labrax) allowed us to identify parvalbumin as a protein marker for differentiation. Further analysis of exudates composition could be a good way to find other indicators. The lysosomal alpha-glucosodase activity is already used to differentiate fresh versus frozen/thawed fillets. Two new indicators were studied : concentration of the nucleotides and their derivatives and free calcium concentration. The total protein and the parvalbumin concentrations were also measured. An Ascendant Hierarchical Clustering (AHC) was done to aggregate the variables into three dissimular clusters : the cellular lysis indicators, the proteins damages indicators and the nucleotides alteration.

Roads and highways in cold regions are negatively affected by settlement of embankments in areas of degraded or degrading permafrost, particularly in areas with mean annual temperatures close to 0°C where permafrost is locally discontinuous. Climate warming and human activities can lead to increases in the temperature of permafrost and to thawing. In regions of discontinuous permafrost, thawing may produce thickening of the active layer, large settlements and non-recoverable shear deformations. These can cause potentially dangerous trafficability issues. The research program involved a test site on Provincial Road PR391, about 18 km northwest of Thompson, Manitoba. The foundation material of PR391 is currently classified as “degraded (thawed) permafrost”. The purpose of this research was to investigate and understand the performance of highway embankments on thawed permafrost. The research involved field instrumentation and data collection, laboratory testing, thermal modelling and frost heave predictions. The results of the field instrumentation and data collection show net heat flow into the ground, as well as development of cyclic seasonal gradients of total head. This is believed to be a significant original contribution to understanding the effects of climate change on highway infrastructure. The results also show that displacements observed at the PR391 embankment are a combination of consolidation and shearing strain of the foundation material. Most studies of embankments concentrate on vertical settlements. This research shows that horizontal movements are also present, a contribution that helps explain the mechanism of lateral spreading. The results of the laboratory testing show that the mechanical and shearing properties of the soil collected from the PR391 embankment are not significantly affected by differences in temperature once water in the soil has actually thawed. The results of the thermal modelling show reasonable trends in simulated ground temperatures compared with the data obtained from the thermistors underneath the embankment. The frost heave prediction of PR391 shows that in situ frost heave characteristics can be estimated by applying the Segregation Potential (SP) frost heave method in field conditions. This provides a valuable field study to the limited number of such studies of Segregation Potential, which are normally done under laboratory conditions.

Roads and highways in cold regions are negatively affected by settlement of embankments in areas of degraded or degrading permafrost, particularly in areas with mean annual temperatures close to 0°C where permafrost is locally discontinuous. Climate warming and human activities can lead to increases in the temperature of permafrost and to thawing. In regions of discontinuous permafrost, thawing may produce thickening of the active layer, large settlements and non-recoverable shear deformations. These can cause potentially dangerous trafficability issues. The research program involved a test site on Provincial Road PR391, about 18 km northwest of Thompson, Manitoba. The foundation material of PR391 is currently classified as “degraded (thawed) permafrost”. The purpose of this research was to investigate and understand the performance of highway embankments on thawed permafrost. The research involved field instrumentation and data collection, laboratory testing, thermal modelling and frost heave predictions. The results of the field instrumentation and data collection show net heat flow into the ground, as well as development of cyclic seasonal gradients of total head. This is believed to be a significant original contribution to understanding the effects of climate change on highway infrastructure. The results also show that displacements observed at the PR391 embankment are a combination of consolidation and shearing strain of the foundation material. Most studies of embankments concentrate on vertical settlements. This research shows that horizontal movements are also present, a contribution that helps explain the mechanism of lateral spreading. The results of the laboratory testing show that the mechanical and shearing properties of the soil collected from the PR391 embankment are not significantly affected by differences in temperature once water in the soil has actually thawed. The results of the thermal modelling show reasonable trends in simulated ground temperatures compared with the data obtained from the thermistors underneath the embankment. The frost heave prediction of PR391 shows that in situ frost heave characteristics can be estimated by applying the Segregation Potential (SP) frost heave method in field conditions. This provides a valuable field study to the limited number of such studies of Segregation Potential, which are normally done under laboratory conditions.

The aim of the current study was to determine if frozen-thawed bull semen can be treated with the acidic buffer MES (2-[N- morpholino] ethanesulfonic acid) without any detrimental effect on the motility, plasma membrane, acrosomal membrane and longevity of sperm. Frozen bull semen was obtained from a local co-operative. The semen was frozen in 0.25 mL French straws at a concentration of 80 x 106 sperm cells per millilitre. Semen of two different batches from ten bulls of four different breeds was used in this study. Three frozen semen straws of each batch were thawed at 38° C for 25 seconds. The thawed semen was pooled and then split into two aliquots. The one aliquot was used as control, whilst the other was exposed to MES treatment. The motility, plasma membrane integrity, acrosomal membrane integrity and longevity of sperm were evaluated. The effect of MES on motility was minimal as only the percentage of aberrantly motile sperm increased two hours after treatment. Although no effect on the plasma membranes were observed, it can be assumed that some damage did occur due to the fact that the acrosomal membranes were affected significantly. No significant effect was found for longevity of sperm between the control and treated samples, but a significant effect was found for both the control and treated samples over time. Although the detrimental effects caused by MES treatment would render some sperm unable to fertilise an oocyte, it is likely that a sufficient portion of sperm would survive the treatment. It is probable that this treatment would also be effective in frozen-thawed buffalo semen. The following step would be to treat semen of footand-mouth disease positive bulls with MES to establish if treatment with MES will be effective in inactivating foot-and-mouth disease virus in semen of infected bulls. Copyright / Dissertation (MSc (Veterinary Science))--University of Pretoria, 2008. / Production Animal Studies / unrestricted

Abstract Fertility declines with advancing age and the number of couples seeking infertility treatment at an older age is constantly increasing. A top quality embryo is believed to have the highest potential for implantation and development into a child. A better understanding of the relative importance of patient and treatment characteristics and of embryo quality could help to optimise the existing therapeutic schemes and the safety of in vitro fertilisation/intracytoplasmic sperm injection (IVF/ICSI). In this work, databases of five Finnish infertility clinics were studied retrospectively. Data on treatments performed in the years 1994–2005 were collected. A total of 19,000 treatment cycles were analysed. Special attention was paid to the relative significance of the transfer of top quality embryos with regards to pregnancy, miscarriage, live birth and cost of treatment in the general IVF/ICSI patient population and in groups with expected poor outcome. The results showed that the transfer of a top quality embryo is associated with a better chance of pregnancy and live birth. However, it does not diminish the probability of miscarriage. Both low and high BMI increase the miscarriage rate. Advancing age and a positive history of previous miscarriages are also associated with a higher probability of miscarriage. In addition, the need for hormonal substitution in cases of frozen-embryo transfer is a risk factor of miscarriage, probably because of suboptimal endometrial function. Since the transfer of several embryos leads to multiple pregnancies, which are associated with a high risk of maternal and fetal complications, elective single embryo transfer (eSET) of a top quality embryo allows all additional good quality embryos to be frozen and transferred later in frozen-thawed embryo transfer cycles. The present work demonstrates that eSET is a safe treatment strategy at least until the age of 40. However, it might not be performed in women with fewer than four collected oocytes, since the prognosis might remain poor even if the response is improved in a following cycle. When eSET is applied routinely and on a large scale, it diminishes treatment costs while increasing the number of deliveries occurring at term, making IVF/ICSI at the same time safer and more affordable even to patients without access to reimbursed IVF treatment.