Dissertations / Theses on the topic 'Fertility of animals'

Post-translational modification is an intracellular process that modifies the properties of proteins to extend the range of protein function without spending energy in de novo peptide synthesis. There are many post-translational modifications, for example, phosphorylation, ubiquitination, and S-nitrosylation. S-Nitrosylation is a post-translational modification which adds nitric oxide (NO) to sulfhydryl groups at cysteine residues to form S-nitrosothiol (SNO), and is required for plant immunity and fertility. Cellular NO changes between a pool of free NO and bound SNO. During pathogen infection, nitrosative stress in plants is mainly controlled by Snitrosothiolglutathione reductase (GSNOR) via the decomposition of GSNO. GSNOR is an alcohol dehydrogenase type 3 (ADH3) which has both GSNOR and formaldehyde dehydrogenase (FDH) activities. The roles of S-nitrosylation in mammals overlap with those in plants. This conservation led us to explore the relationship between S-nitrosylation, immune response, and fertility in Drosophila melanogaster as it might prove to be a good genetic model for further analysis of the role of S-nitrosylation in animals. I have identified fdh as the likely gsnor in D. melanogaster and have knocked this out using an overlapping deficiency technique in order to observe the effect on immunity and fertility. There are two main pathways in the Drosophila innate immune response, the Toll pathway for protecting against gram-positive bacteria and fungi, and the Imd pathway against gram-negative bacteria. I have investigated the effect of removing GSNOR on sensitivity to gramnegative bacteria (Escherichia coli and Erwinia carotovora) by septic and oral infection, and to fungi (Beauveria bassiana). Susceptibility to infection by the gram negative bacteria was similar to wild-type but susceptibility to B. bassiana was increased. This increase in susceptibility correlated with reduced anti-fungal antimicrobial peptide (AMP) production after B. bassiana infection. This suggests that GSNOR might be required for the normal activity of the Toll pathway or novel Toll-independent processes. We also observed that gsnor knockout impairs fertility and development of embryos.

Thesis (Ph.D.)--Kent State University, 2009.
Title from PDF t.p. (viewed May 17, 2010). Advisor: Srinivasan Vijayaraghavan. Keywords: sperm; spermatogenesis; PP1gamma2; PP1gamma1; mice; transgene. Includes bibliographical references (p. 102-123).

Thesis (M. Tech.) -- Central University of Technology, Free State, 2007
Four different South African indigenous (Naked Neck (NN), Ovambo (OVB), Venda (VD) and Potchefstroom Koekoek (PK) chicken breeds were used in this study. From each of the four breeds of chicken, 40 hens and 8 cocks were selected randomly. Two groups each of sixteen cocks were subsequently formed: high performing (HP) and low performing (LP) groups to determine the relationships between cock semen viability and the fertility of artificially inseminated South African indigenous layer breeds. Semen was collected following five minutes of sexual massage (5SM) and evaluated for semen volume (ml), sperm motility (%), live sperm (%) and total sperm (x109/ml). Semen from each cock was then used to inseminate five hens per breed, in each treatment. Each hen was inseminated twice a week throughout the duration of the trial. During the experimental period, each hen was inseminated with 0.05 ml diluted semen. The artificially inseminated hens were examined for average egg weight (g), fertility (%), hatchability of set eggs (%), live chicks (%), normal chicks (%) and chick weight (g). A total of 1600 eggs, i.e. 400 eggs from each breed were collected in three batches following artificial insemination from individually caged hens and were hatched to compare hatching parameters among breeds. The hatchability traits of hens of the four breeds (NN, OVB, PK and VD) were compared. Hatching egg weight had significant (P < 0.05) difference among the four breeds. The results of this study indicate that semen viability exemplified by ejaculate volume, sperm motility; live sperm and total sperm per ejaculate were significantly (P < 0.01) superior in the HP cocks compared to the LP cocks. Hens inseminated with semen from the HP cocks in each experimental group resulted in higher egg weight (g), fertility (%), hatchability of set eggs (%), live chicks (%), normal chicks (%) and chick weight (g). Significant positive relationships existed between semen volume and sperm motility (P < 0.05), semen volume and live sperm cells (P < 0.01), semen volume and total sperm (P < 0.01) in NN, OVB and VD, with negative correlations in PK. Some positive correlations were found between sperm motility and live spermatozoa (P < 0.01), sperm motility and total sperm (P < 0.01), live sperm and total sperm (P< 0.01) in NN, OVB, PK and VND. Fertility was the highest in the HP group. Fertility was also the highest in PK, intermediate and similar in OVB and NN and lowest in VD (P<0.05). Breed had a significant effect on hatchability of fertile eggs (P<0.05). Hatchability of total eggs set was highest in PK and NN, intermediate in OVB and lowest in VD (P<0.05). Breed had a significant effect on live, normal chicks and chick weight (P<0.05). Live chick was the highest in NN, whereas at day-old, normal chick and chick weight at hatching were the highest (23.50 ± 0.11) (P<0.05) in PK (98.14 ± 0.67 vs. 37.90 ± 0.28 g), intermediate and similar in NN (87.90 ± 0.63 vs. 23.50 ± 0.11) and OVB (87.75 ± 0.45 vs. 32.81 ± 0.49 g) and the lowest but with an acceptable value in VD (76.85 ± 0.46 vs. 26.90 ± 0.36 g). There were some correlations among different hatchability traits depending on breed. The correlations were more profound among PK. It was clear that chick weight as percent of egg weight was not just a function of egg weight, and that genotype also played an important role favouring the heavier breeds. The results obtained in this study on the relationships between cock semen viability and the fertility of artificially inseminated South African indigenous layer breeds elucidate that the use of high performing (HP) cocks following five minutes of sexual massage, prior to semen collection and artificial insemination of layers is a practical method for optimising sperm viability and subsequent fertility of hens. The results of this study suggest that the Potchefstroom Koekoek (PK) cocks and hens are superior to the Naked Necks (NN), Ovambo (OVB) and Venda (VD). The Ovambo and Naked Neck cocks ranked second in donating quality semen as well as in improving the fertility and hatchability traits of the indigenous chicken breeds. Thus selection of high performing cocks through five minutes sexual massage prior to semen collection and use is recommended for poultry AI breeding programmes.

Orientador: Wilma De Grava Kempinas
Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
Made available in DSpace on 2018-08-18T02:46:53Z (GMT). No. of bitstreams: 1 Fernandez_CarlaDalBianco_D.pdf: 4399629 bytes, checksum: 5e3c4d594df1ba1dd65fdfd541b7fa14 (MD5) Previous issue date: 2011
Resumo: O sobrepeso e a obesidade estão aumentando rapidamente, tornando-se uma epidemia mundial. Estas duas condições podem ser definidas simplesmente como o acúmulo excessivo de gordura nos adipócitos, células do tecido adiposo responsáveis pela síntese e liberação de leptina, um hormônio protéico cuja principal ação é inibir o apetite e estimular o gasto energético, mas que também atua na regulação do eixo hipotalâmico-hipofisáriogonadal. As concentrações séricas de leptina apresentam-se elevadas, proporcionalmente à quantidade de gordura, em muitos modelos animais de obesidade e na obesidade humana. Recentemente alguns estudos têm mostrado a relação entre obesidade e infertilidade masculina, mas até o momento os resultados são controversos. Da mesma forma, o papel da leptina no sistema reprodutor masculino não está muito esclarecido, principalmente quando este hormônio está em excesso, como na obesidade. Assim, os objetivos do presente estudo foram investigar os efeitos da obesidade induzida por consumo de dieta rica em gordura em parâmetros reprodutivos de ratos machos. Também foi investigado o papel da leptina na fertilidade e em alguns parâmetros espermáticos, em ratos machos adultos. Para isso, ratos machos da variedade Wistar foram alimentados com dieta hipercalórica rica em gordura (DH) ou com dieta padrão (DP) durante 15, 30 ou 45 semanas, e foram avaliados quanto ao índice de obesidade, níveis séricos de leptina, peso dos órgãos reprodutores e contagens espermáticas. Em um segundo experimento, os animais receberam as diferentes dietas apenas por 15 semanas, tempo suficiente para provocar obesidade. Esses animais foram avaliados quanto às dosagens dos hormônios sexuais, comportamento sexual, motilidade e morfologia espermática, fertilidade após cruzamento natural e inseminação artificial in utero. Além disso, ratos machos adultos não-obesos foram tratados com leptina exógena, ou com salina, ip, por 42 dias e avaliados quanto ao peso dos órgãos reprodutores, contagens espermáticas, motilidade dos espermatozóides e fertilidade após inseminação artificial. Após 15, 30 ou 45 semanas, os animais alimentados com DH apresentaram um aumento significativo no índice de obesidade e nos níveis de leptina. O peso dos órgãos reprodutores e as contagens espermáticas foram similares entre os dois grupos, em todos os períodos estudados. Nos animais que receberam DH por 15 semanas, o comportamento sexual e a fertilidade após acasalamento natural foram semelhantes aos do grupo controle. Da mesma forma, os níveis de testosterona não foram alterados, mas os de estradiol aumentaram nos animais que ingeriram DH. Além disso, a qualidade espermática foi reduzida nestes animais, como evidenciado pela baixa porcentagem de espermatozóides com movimento progressivo. A alteração da motilidade espermática foi seguida por uma redução quantitativa do potencial de fertilidade após inseminação artificial in utero. Os animais tratados com leptina não apresentaram diferenças nas contagens espermáticas nem na motilidade quando comparados aos animais controle. No entanto, o potencial de fertilidade reduziu cerca de 40% e a taxa de perdas pré-implantação foi duas vezes maior nos animais tratados com leptina. Juntos, esses resultados indicam que a obesidade diminuiu a qualidade espermática, prejudicando a fertilidade dos ratos machos, e que, provavelmente a leptina está relacionada a este prejuízo
Abstract: Overweight and obesity are rapidly becoming a worldwide epidemic that affects children and adults. These two statuses are often defined simply as a condition of abnormal or excessive fat accumulation in adipose tissue, whose cells are responsible for synthesis and release of leptin. This is a protein hormone whose main action is to decrease appetite and increase energy expenditure, but it also act as regulatory signal for gonadal axis, among other functions. Serum leptin concentration is augmented in several obesity animal models and in obese human, proportionately to the amount of fat. Some studies have shown a relationship between obesity and male infertility, but until now it remains controversial. In the same way, the role of leptin on male reproductive function is not clear, mainly when the hormone is in excess as occur in obesity. Thus, the aims of the present study were to investigate the effects of high-fat diet-induced obesity on male rats' reproductive parameters. Furthermore it was evaluated the possible role of the excess of leptin on fertility and some sperm parameters in adult male rats. To achieve these aims, male Wistar rats were fed a high-fat diet (HFD) or standard chow (SD) for 15, 30 or 45 and were evaluated by adiposity index, serum leptin levels, reproductive organ weights and sperm counts. In a second experiment rats received HFD or SD only for 15 weeks, long enough to cause obesity. Sexual hormones, sexual behavior, sperm morphology and motility, fertility after natural mating and after artificial in utero insemination were evaluated. Moreover, non-obese adult male rats were treated with exogenous leptin ip (30?g/kg/day) or vehicle for 42 days and were evaluated for sperm count in testis and epididymis, sperm motility and fertility after in utero artificial insemination. After 15, 30 or 45 weeks, HFD-fed animals presented significant increases in obesity index and serum leptin levels. Reproductive organ weights and sperm counts in the testis and epididymis were similar between the two groups at all timepoints studied. In HFD-fed animals only for 15 weeks, sexual behavior and fertility after natural mating were not altered by the diet regimen. In these animals, intergroup testosterone levels were also comparable, but estradiol levels were increased in HDF rats. Furthermore, sperm quality was reduced in 15 weeks HFD-fed animals as evidenced by their decreased percentage of sperm with progressive movement. This altered motility parameter was followed by a quantitative reduction in fertility potential after artificial in utero insemination. Leptin treated animals did not show any differences in sperm counts either in the testis or epididymis. In the same way, the percentage of progressive, non-progressive and immotile sperm was similar between the two groups. However, the fertility potential showed a reduction of about 40% whereas the pre-implantation loss rate increased more than two fold in leptin treated animals. These results indicate that obesity lead to a decrease in sperm quality, impairing male fertility and probably the excess of leptin is related with this alteration of fertility
Doutorado
Biologia Celular
Doutor em Biologia Celular e Estrutural

Proteins in bull semen have been found that increase fertility, including the fertility-associated antigen (FAA). The uses of FAA-positive bulls have been found in previous studies to increase conception rates at least 16%. Increased fertility rates by FAA-positive bulls increased the number of pregnant cows early in the breeding seasonUsing historical data to compute an equation that can demonstrate buyers and producers in the Southwestern United States the benefits of using and testing for FAA-positive bulls over time. The cost for addition of a cassette to the BSE testing will be offset by an increase in conception rate, birthing rate, and increase calf weaning weightsThis study focuses on the Southwestern United States, but eventually the information may be used nationwide. Fertility-associated antigen is a new science that is being studied to help the cattle industry increase production and profitability.

These studies were conducted to determine the chemical identity of heparin binding proteins in semen that are related to fertility of bulls. The first study describes the isolation and identification of a 31,000 dalton fertility-associated antigen (FAA). FAA was found to have significant primary structure homology to a recently described novel DNase I-like protein. The physiological significance of the similarities between FAA and a protein homologous to DNase I is unknown at this time as no function has been described for the DNase I-like protein. The second study describes the isolation and identification of a 24,000 dalton seminal heparin binding protein. It was found to be similar, if not identical, to tissue inhibitor of metalloproteinase-2 (TIMP-2). TIMP-2 regulates matrix metalloprotease activity and therefore, potentially plays a key role in the structural makeup of the extracellular matrix. These findings suggest that regulation of enzymatic activity in seminal fluid is in large part a function of heparin binding proteins that have been correlated to fertility of bulls, one being a potentially novel extracellular nuclease and a second acting as a specific inhibitor of metalloprotease activity. In addition to the ability to modulate capacitation of sperm, seminal heparin binding proteins likely are key players in protecting sperm and male reproductive tract tissues from enzymatic hydrolysis. The proteins identified in this dissertation represent novel additions to the previously described seminal heparin binding protein families. Clearly, these data indicate a growing complexity of seminal fluid and implicate a novel Dnase I-like protein and TIMP-2 in affecting cellular events related to fertility potential of males.

MicroRNAs are small RNA molecules that could possibly play a major role in fertility. In the experiment, spermatozoa were extracted from bovine followed by an extraction of total RNA. Bovine spermatozoa were extracted from two bulls of different fertility, high and low fertility. An expression array was done to compare the expression levels of the microRNAs. It was shown that thousands of microRNAs are present in bovine spermatozoa but only a small amount was significantly expressed. The microRNAs from low fertility bulls were more highly expressed than those in high fertility bulls. A Bioanalyzer gel was used to confirm the results of the microarray data. The microRNAs were present in the bulls spermatozoa at 25 nucleotides. The functions of the significantly expressed microRNAs are not known but there is a great possibility that their functions affect fertility.

La vaca lletera d’alta producció actual pateix un estrés metabòlic ques’uneix a altres tipus d’estrés com el del part i de la gestació. Malgratles millores en nutrició i genètica, la eficàcia reproductiva d’aquestsanimals ha anat disminuint any rere any. Per aquest motiu, l’objectiud’aquesta tesi es establir un protocol diagnòstic i terapèutic durant els dies 15-21 postpart que identifiqui els animals amb endometritis i millori la recuperació postpart de vaques lleteres d’alta producció. Alhora, es planteja un protocol desincronització basat en progesterona intravaginal aplicat juntament ambgonadotropina coriònica equina. Aquest protocol pretén millorar laeficàcia reproductiva de la vaca lletera d’alta producció, amb elevat nombre d’anestre postpart.
La vaca lechera de alta producción actual sufre un estrésmetabólico quese une a otrostipos de estrés, como pueden ser el parto o la gestación. Apesar de las mejoras en nutrición y genética de los últimosaños, laeficacia reproductiva de estos animales ha disminuidoconsiderablemente. Por este motivo el objetivo de esta tesis es establecer un protocolodiagnóstico y terapéuticodurante los días 15-21 postparto que identifique los animales con endometritis y mejore la recuperaciónpostparto de las vacaslecheras de alta producción. A suvez se plantea un protocolo de sincronizaciónbasado en progesteronaintravaginalimplementado con gonadotropina coriónica equina (eCG). Esteprotocolopretendemejorareficaciareproductiva de la vaca lechera dealta producción, especialmente de las vacasanéstricas.
Nowadays, high producing dairy cow suffers a metabolic stress that may be associated to other stress such as parturition or gestation.Despite the nutritional and genetic improvements in last decades,reproductive performance has dramatically decreased. For that reason, the aims of this thesis are to implement a diagnostic and therapeutic protocol during Days 15-21 postpartum that detect cows suffering from endometritis and to improve postpartum involution and subsequent reproductive performance in high producing dairy cows. Moreover, a progesterone based synchronization protocol plus equine chorionic gonadotrophin (eCG) is tested. The objective of this protocol is to improve the reproductive performance of high producing dairy cows, especially anestric cows.

La vaca lletera d’alta producció actual pateix un estrés metabòlic ques’uneix a altres tipus d’estrés com el del part i de la gestació. Malgratles millores en nutrició i genètica, la eficàcia reproductiva d’aquestsanimals ha anat disminuint any rere any. Per aquest motiu, l’objectiud’aquesta tesi es establir un protocol diagnòstic i terapèutic durant els dies 15-21 postpart que identifiqui els animals amb endometritis i millori la recuperació postpart de vaques lleteres d’alta producció. Alhora, es planteja un protocol desincronització basat en progesterona intravaginal aplicat juntament ambgonadotropina coriònica equina. Aquest protocol pretén millorar laeficàcia reproductiva de la vaca lletera d’alta producció, amb elevat nombre d’anestre postpart.
La vaca lechera de alta producción actual sufre un estrésmetabólico quese une a otrostipos de estrés, como pueden ser el parto o la gestación. Apesar de las mejoras en nutrición y genética de los últimosaños, laeficacia reproductiva de estos animales ha disminuidoconsiderablemente. Por este motivo el objetivo de esta tesis es establecer un protocolodiagnóstico y terapéuticodurante los días 15-21 postparto que identifique los animales con endometritis y mejore la recuperaciónpostparto de las vacaslecheras de alta producción. A suvez se plantea un protocolo de sincronizaciónbasado en progesteronaintravaginalimplementado con gonadotropina coriónica equina (eCG). Esteprotocolopretendemejorareficaciareproductiva de la vaca lechera dealta producción, especialmente de las vacasanéstricas.
Nowadays, high producing dairy cow suffers a metabolic stress that may be associated to other stress such as parturition or gestation.Despite the nutritional and genetic improvements in last decades,reproductive performance has dramatically decreased. For that reason, the aims of this thesis are to implement a diagnostic and therapeutic protocol during Days 15-21 postpartum that detect cows suffering from endometritis and to improve postpartum involution and subsequent reproductive performance in high producing dairy cows. Moreover, a progesterone based synchronization protocol plus equine chorionic gonadotrophin (eCG) is tested. The objective of this protocol is to improve the reproductive performance of high producing dairy cows, especially anestric cows.

Sulfogalactosylglycerolipid (SGG) is a sulfoglycolipid present specifically at a substantial level in mammalian male germ cells. It has been shown to function as an adhesion molecule important for sperm-egg interaction and a structural lipid involved in formation of sperm lipid rafts during capacitation in vitro. Due to the unique characteristics and functions, SGG can potentially serve as a biomarker for sperm fertility as well as a target for development of a non-hormonal contraceptive. To confirm the in vivo roles of SGG, we sought for transgenic mice with reduced amounts of sperm SGG. Cgt knockout male mice, transgenetically deficient in UDP-galactose:ceramide galactosyltransferase (CGT), an enzyme involved in SGG synthesis, are infertile due to spermatogenesis disruption. However, the Cgt+/- males can still produce sperm and sire offspring. We hypothesized that Cgt+/- males, expected to have reduced SGG amounts, would have compromised fertilizing ability and could serve as in vivo models for studying roles of SGG in fertilization and spermatogenesis. Unexpectedly, our results revealed that Cgt+/- males exhibited unimpaired spermatogenesis and fecundity. Moreover, the levels of SGG as well as lipid profiles of sperm and testes of Cgt+/- mice were similar to those of the wild type, suggesting that compensatory mechanisms must have occurred to maintain SGG levels in the Cgt+/- mice. Although these results revealed that Cgt+/- mice could not be used as the animal models, they implicated significance of normal testis and sperm SGG levels in maintaining normal spermatogenesis and fertility. The possible compensatory mechanisms regulating SGG levels were further investigated in Cgt+/- mice. As expected, only half of Cgt mRNA expression level of the wild type was transcribed in the Cgt+/- testes; however, testicular CGT polypeptides as well as their enzymatic activities in the Cgt+/- mice were found at a comparable level to those of the wild type. On the other hand, no change was found in terms of mRNA levels, polypeptide levels or enzymatic activities of arylsulfatase A (ASA), the enzyme responsible for SGG degradation in the testis. In conclusion, the compensatory mechanisms for SGG level adjustment in Cgt +/- mice occurred through the biosynthetic pathway, rather than the degradation pathway, by increasing the CGT polypeptide expression level. Therefore, identification of specific spermatogenic cell stages, contributing to normal expression levels of CGT and SGG in the Cgt+/- testes warrants further studies, as these studies should provide useful information regarding CGT and SGG importance during male germ cell development. In addition, a new approach to produce the animal models that can produce sperm with reduced SGG levels should be attempted. The RNA interference (RNAi) techniques may be tried to achieve this goal.

Routinely collected herd management data in a variety of formats were collated from 468 dairy herds, and novel objective measures of data recording quality were developed and applied. This revealed that there was a substantial amount of variation in data quality between herds, and the vast majority of herds failed to meet the threshold level for at least one of the data quality measures used. Analysis of trends in reproductive performance across the herds with good quality fertility event recording suggested that their fertility was generally declining through the first half of the 2000s, but there was some evidence that improvements in submission rate were beginning to reverse this decline in the later years studied (up to 2007). Associations between reproduction and two endemic diseases common in dairy cattle (mastitis and lameness) were explored using multilevel discrete time survival modelling, and probabilistic sensitivity analysis (PSA) used to contextualise and illustrate the results. In both cases, statistical modelling revealed significant and sizeable associations between disease events and reproductive outcomes at lactation level. However, simulation and application of PSA showed that a herd’s incidence rate of either disease was highly unlikely to influence its overall reproductive performance to a clinically relevant degree when other inputs to herd fertility were also considered. Factors associated with the proportion of serves leading to a pregnancy (pregnancy rate) were explored using multilevel logistic regression modelling. This revealed that relatively little of the variation in herd pregnancy rate is explainable by routinely recorded milk recording data (including constituent concentration in early lactation as well as daily and lactation yields). A large amount of the unexplained variation was revealed to be at herd level and very little at cow level, suggesting that investigation of herd management practices associated with pregnancy rate would be rewarding.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
A obstruÃÃo das artÃrias uterinas promove isquemia e/ou necrose no Ãtero, no entanto nÃo se conhece com que intensidade essas lesÃes ocorrem. Os objetivos deste estudo sÃo: descrever e padronizar uma tÃcnica de ligadura da artÃria uterina (LAU) direita em ratas nÃo grÃvidas e avaliar os efeitos deste modelo em Ãteros e ovÃrios de ratas. Estudo experimental, utilizando 64 ratos, 48 fÃmeas e 16 machos (Rattus norvergicus, variedade albina) maduros, de fertilidade comprovada. As ratas foram alocadas aleatoriamente em 8 grupos, de seis indivÃduos. Quatro grupos foram submetidos à tÃcnica de LAU direita e sacrificados nos dias 1, 7, 14 e 21 apÃs o procedimento. Outros 3 grupos foram acasalados nos dias 1, 7 e 14 apÃs a LAU, e comparados com o grupo controle quanto à fertilidade. ApÃs o sacrifÃcio, eram retirados, para anÃlise histopatolÃgica, os ovÃrios, e os hemi-Ãteros. Realizou-se anÃlise histolÃgica, que avaliou o Ãtero quanto à congestÃo, hemorragia, edema intersticial e perda de coesÃo celular atravÃs de um escore que varia de 0 a 3. Os ovÃrios foram avaliados de acordo com o nÃmero de folÃculos em desenvolvimento e corpos lÃteos. Para a anÃlise estatÃstica foi utilizado o software SPSS (Statistical Package for Social Sciences) versÃo 13.0, p < 0,05 foi considerado estatisticamente significativo. O projeto de pesquisa foi enviado para a ComissÃo de Ãtica em Pesquisa Animal (CEPA) da Universidade Federal do CearÃ, com protocolo de nÃmero 90/08. O modelo da tÃcnica foi realizado em ratas nÃo-grÃvidas utilizando a ligadura na porÃÃo inferior da artÃria uterina direita, mantendo o hemi-Ãtero esquerdo (HUE) como controle. Em nenhum momento dos dias de sacrifÃcio, apÃs a LAU (1, 7, 14 e 21 dias), houve diferenÃa entre os escores histolÃgicos de isquemia dos hemi-Ãteros direitos (HUD) distais e proximais (p > 0,05). Os escores histolÃgicos de isquemia dos hemi-Ãteros direitos no decorrer do tempo aumentaram substancialmente a partir do 7 dia (p=0,003). Da mesma maneira ocorreu com os hemi-Ãteros esquerdos (p=0,001). A Ãnica diferenÃa significativa observada na comparaÃÃo dos escores dos hemi-Ãteros direito e esquerdo ocorreu no 1 dia apÃs LAU (p=0,026), à custa de congestÃo. Os ovÃrios esquerdos nÃo apresentaram alteraÃÃes no nÃmero de folÃculos e de corpos lÃteos apÃs LAU e, os ovÃrios direitos apresentaram nÃmero de folÃculos e corpos lÃteos semelhante ao controle a partir do 21 dia da LAU. A percentagem de ratas grÃvidas que foram submetidas a LAU foi de 44,4%, comparado com 100% das ratas controle que engravidaram (p=0,024). Observou-se ainda uma reduÃÃo na mÃdia do nÃmero de fetos por rata (p=0,029). Pode-se concluir que o modelo estabelecido foi efetivo e de fÃcil reprodutibilidade, bem como as alteraÃÃes histolÃgicas encontradas no Ãtero ocorrem de forma discreta. Em relaÃÃo a funÃÃo ovulatÃria, pode-se dizer que o nÃmero de folÃculos e corpos lÃteos dos ovÃrios direitos, a partir do 21 dia, permaneceram semelhantes aos dos esquerdos (controle). A fertilidade, porÃm, mostrou-se reduzida apÃs o estabelecimento desta tÃcnica.
The obstruction of the uterine arteries promotes ischemia or necrosis in the uterus. However it is not known how strongly these injuries occur. The objectives of this study are to describe and standardize a technique of right uterine artery ligation (UAL) in non-pregnant rats and to evaluate the effects of this model in the uteri and ovaries of female rats. An experimental study using 64 rats, 48 females and 16 males (Rattus norvegicus, albino variety) mature, with proven fertility. The rats were randomly allocated into 8 groups of six individuals. Four groups were subjected to the technique of right UAL and sacrificed on days 1, 7, 14 and 21 after the procedure. Other 3 groups were mated on days 1, 7 and 14 after the UAL, and compared with the control group regarding fertility. After sacrifice, ovaries and the hemi-uteri, were removed for histological analysis wich was carried out histological analysis, which evaluated the uteri and the congestion, hemorrhage, interstitial edema and loss of cell cohesion through a score ranging from 0 to 3. Ovaries were evaluated according to the number of follicles and corpora lutea. For statistical analysis we used SPSS software (Statistical Package for Social Sciences) version 13.0, p <0.05 was considered statistically significant. The research project was submitted to the Ethics Committee on Animal Research (ECAR) Federal University of CearÃ, under the number 90/08. The model of the technique was performed in non-pregnant rats using ligation in the lower portion of the right uterine artery, keeping the left hemi-uteri (LHU) as control. At no time during periods of ischemia established (1, 7, 14 and 21 days) there was any difference between the histological scores of ischemia of the right hemi-uteri (RHU) distal and proximal (p> 0.05). The histological scores of ischemia of the right hemi-uteri over time increased significantly from day 7 (p = 0.003), just as occurred with the left hemi-uteri (p = 0.001). The only significant difference observed when comparing the scores of the right hemi-uteri and left occurred on day 1 after UAL (p = 0.026), due to congestion. The left ovary showed no changes in the number of follicles and corpora lutea after UAL, and the right ovary showed a number of follicles and corpora lutea similar to control from the 21th day of UAL. The percentage of pregnant rats that were subjected to ischemia was 44.4%, compared with 100% of control rats that became pregnant (p = 0.024). There was also a reduction in the average number of fetuses per mother (p = 0.029). It can be concluded that the model established was effective and highly reproducible, and histological changes found in the uteri are mild. As to ovulatory function, one can say that the number of follicles and corpora lutea of the right ovaries, after 21 days, remained similar to the left (control). Fertility, however, was reduced after the establishment of this technique.

Seulement 30% des cas d'infertilité masculine peuvent être diagnostiqués grâce à un spermogramme. Comme l'épididyme joue un rôle important dans la maturation posttesticulaire spermatique, le dysfonctionnement de cet organe peut être à l'origine de cas d'infertilité non expliquée. Ainsi, puisque la communication intercellulaire au niveau du tractus reproducteur mâle est différente lorsqu’il y a un problème de fertilité, nous avons émis l’hypothèse qu’une signature moléculaire précise pourrait aider à mieux comprendre et diagnostiquer une plus large population. Pour ce faire, nous avons utilisé un modèle murin génétiquement modifié ayant une délétion conditionnelle (cKO) pour une enzyme impliquée dans la biogenèse des microARNs (miARNs) au niveau proximal de l’épididyme. Nous avons comparé le profil d’expression des miARNs et des gènes au sein de l’épididyme avec un modèle de souris témoin. Cent cinquante-quatre miARNs ont illustrés une forte diminution d’expression chez la souris génétiquement modifiée, dont miR-210, miR-672, miR-191 and miR-204. Notons également la variation d’expression des protéines AZGP1 (Zn-alpha 2-glycoprotein) et PATE4 (Prostate And Testis Expressed 4 protein) qui ont particulièrement attiré notre attention, car elles sont directement impliquées dans le pouvoir fécondant masculin. Deuxièmement, comme ces molécules se sont avérées être relarguées dans le milieu extracellulaire, nous avons décidé d’investiguer leur mode de sécrétion dans le liquide épididymaire afin d’évaluer leur potentiel en tant que biomarqueurs pour un certain type d’infertilité masculine. En nous référant aux critères de l’Organisation Mondial de la santé (OMS), j’ai créé une biobanque contenant 43 échantillons humains. J’ai pu mettre également en pratique une méthode plus actuelle d’isolement de vésicules membranaire à partir de fluide séminal. Finalement, j’ai validé l’association de 4 molécules (miR-672, miR- 191 and miR-204 et AZGP1) avec les exosomes. Tous ces résultats suggèrent donc qu’il existe un profil moléculaire associé au phénotype de fertilité et que ces nouvelles molécules extracellulaires pourraient aider à représenter des sous-types d’infertilité masculine.
Only 30% cases of male infertility can be diagnosed with a spermogram. As the epididymis plays an important role in spermatic post-testicular maturation, the dysfunction of this organ may be at the origin of unexplained infertility. Thus, since intercellular communication in the male reproductive tract is different when there is a fertility problem, we hypothesized that a precise molecular signature could help to better understand and diagnose a larger population. To do this, we used a genetically modified murine model with a conditional kockout (cKO) for an enzyme involved in the biogenesis of microRNAs (miRNAs) at the proximal level of the epididymis. We compared the epididymal miRNAs and genes expression pattern between a control and cKO mouse model. One hundred and fifty-four miRNAs showed a strong expression decrease in the genetically modified mice, including miR-210, miR-672, miR-191 and miR-204. Note also the variation in expression of the proteins AZGP1 (Zn-alpha 2-glycoprotein) and PATE4 (Prostate And Testis Expressed 4 protein) which have particularly attracted our attention, because they are directly involved in the male fertility. Secondly, since these molecules were found to be released into the extracellular environment, we decided to investigate their secretion pattern in the epididymal fluid in order to evaluate their potential as biomarkers for a certain type of male infertility. Referring to the criteria of the World Health Organization (WHO), I was able to create a biobank containing 43 human samples. I was also able to practice a more current method of isolating membrane vesicles from seminal fluid. Finally, I validated the association of 4 molecules (miR-672, miR-191 and miR-204 and AZGP1) with exosomes. All these results therefore suggest that there is a molecular profile associated with the fertility phenotype and that these new extracellular molecules might help to represent subtypes of male infertility.

During spermatogenesis, histones are largely replaced by transition proteins and protamines in normal stallions. Incomplete nucleoprotein exchange results in the abnormal retention of histones and transition proteins, which is an indicator of poor sperm quality. Equine nucleoprotein exchange has not previously been investigated in detail, so that equine sperm chromatin quality problems, which are often responsible for poor breeding performance of stallions, are not well understood. In order to characterize chromatin remodeling events in stallion spermatogenesis and to identify proteins indicative of sperm chromatin defects, such as excessive amounts of histones, we identified antibodies that recognize equine testis-specific proteins of interest. Immunoblotting of testis and sperm protein lysates and immunofluorescence staining of histological tissue sections were used to identify candidate marker proteins of incomplete sperm chromatin maturation. Results of the study, which represents the first comprehensive characterization of the nucleoprotein exchange during spermatogenesis in the stallion, challenge the paradigm that the main function of histone H4 lysine (hyper-) acetylation (concomitant H4K5 and H4K8 acetylation) is to facilitate nucleosome ejection during spermatid nuclear elongation to allow for transition protein and protamine insertion into the chromatin. That paradigm was based on observations in mice and rats where H4 acetylation in several lysine residues occurs just prior to or during nuclear elongation. In contrast, the equine data presented here show strong acetylation of H4 in K5, K8 and K12 positions immediately after meiosis in round spermatids, independent of nuclear transition protein 1 deposition. Furthermore, results of H4K16 acetylation analyses underline the importance of this mark, which is likely mediated by DNA damage signaling pathways, emphasizing the importance of DNA repair processes for the exchange of nucleoprotein exchange in spermiogenesis and therefore, in extension, for male fertility. In addition, a revised description of the equine spermatogenic cycle is proposed here that is better aligned with human, mouse and rat spermatogenesis. Finally, the testis-specific histone variant TH2B was identified as a potential quantitative marker of equine sperm quality.

Spermatozoa are highly specialised cells which display a range of unique features associated with their crucial function of egg fertilisation. One of the most striking characteristics of spermatozoa, in biochemical terms, is the extremely high proportion of long chain highly polyunsaturated fatty acids present as components of the plasma membrane phospholipids. This high degree of unsaturation is almost unique amongst animal cells; the only other cell types which display similar levels of these polyunsaturates are the neurons of the brain and retina. The reason why spermatozoa exhibit such an unusual fatty acid composition is not clear but it is feasible that the highly unsaturated phospholipids may confer a high degree of flexibility on the sperm plasma membrane as well as provide a potential energy source in order to facilitate the characteristic flagellar motion of these cells. There is also evidence that spermatozoa lipids play a crucial role in the membrane fusion and signal transduction events associated with the acrosome reaction and fertilisation. Initial observations were made between the semen of domestic cockerel and bull with respect to animal ageing and semen quality. Within both species similar patterns were observed in that with age there was a loss of the long chain C20 and C22 polyunsaturated fatty acids accompanied by a loss of the major antioxidant enzyme systems. A decrease in phosphatidyl ethanolamine and an increase of phosphatidyl choline with age were also observed to be associated with a reduction in semen quality parameters and fertility in both species. In contrast cockerels displayed significant increases in spermatozoa and seminal plasma lipid levels where the bull exhibited losses, reflecting possible differences in spermatozoa metabolism and function.

Dissertação de Mestrado Integrado em Medicina Veterinária
Oestrus intensity in Swedish dairy cattle – Is there a relationship with animal-based welfare parameters? A welfare field study was carried out in a Swedish experimental dairy farm to determine the presence and relevance of relationships between animal welfare-based parameters and fertility in heifers and first parity dairy cows with high genetic potential for milk production. The main hypothesis tested was that strong signalling of oestrus is an indicator of good welfare and that it is related to low avoidance distance, low lameness score and acceptable pregnancy rate. A field trial was initiated to validate a commercially automated progesterone measuring instrument (eProCheck®) designed to ease reproduction management. The studied sample included 68 females, balanced in terms of breed and category (Swedish Red or Swedish Holstein; heifer or cow). The animals were included into the study after oestrus detection and artificial insemination (AI) decision by the personnel at the farm. Oestrus intensity (OI) was determined in parallel by the personnel (NCV_OI) and by the author (OBS_OI). On day 0, defined by AI, progesterone relative level was assessed in blood (and milk - cows). On day 7, avoidance distance (at the feeding rack and inside the stable), body condition, lameness and progesterone level were assessed. On day 20, progesterone level was determined, as well as pregnancy through trans-rectal ultrasonography. Pregnancy was confirmed with trans-rectal palpation by day 50. The NCV_OI and OBS_OI differed, but the last revealed to be higher in heifers than in cows. The overall pregnancy rate (55%) was acceptable, but heifers had a higher performance than cows (70% vs. 37%), being higher oestrus intensity, both NCV_OI and OBS_OI, reflected in a higher pregnancy rate. Standing oestrus (high OBS_OI) had 3.8-fold higher odds of pregnancy, compared to the detection based on secondary oestrus signs. Though, AIs based on secondary signs had acceptable pregnancy rates (45 % in low vs. 46 % in medium OI). Strong oestruses (NCV_OI) had 5.3-fold higher odds of pregnancy than weak and clear ones grouped. Non-lame had 4.8-fold higher odds of pregnancy than lame animals. Avoidance distance means were short (<1m), as a reflex of a good human-animal interaction at the farm. Although OBS_OI tended to correlate negatively with avoidance distance at the feeding rack in primiparous cows, the relationships between OI and avoidance distances were found inconclusive and statistically nonsignificant. Results of preliminary eProCheck® trials seem promising regarding early pregnancy diagnosis, due to high sensitivity with a single test on day 20. Although further testing is required, it is considered a valuable complement for on-farm reproductive management. In conclusion, OI and lameness were associated with pregnancy outcome, confirming the importance of these factors in modern dairy farming, although global animal welfare level was considered acceptable within the studied parameters. Yet, re-evaluation of the prevention programme for lameness is needed to address its high prevalence in primiparous cows.
RESUMO - Performance e Bem-estar Animal Intensidade do cio em bovinos de leite na Suécia – Existem relações com os parâmetros de bem-estar baseados no animal? - Foi realizado um estudo de campo numa vacaria experimental de leite na Suécia para determinar a presença e relevância das relações entre parâmetros de bem-estar com base no animal e fertilidade de novilhas e vacas primíparas com alto potencial genético para produção de leite. A principal hipótese testada admite que a sinalização intensa do estro é um bom indicador de bem-estar e que está relacionada com uma curta distância de fuga, um baixo grau de claudicação e uma taxa de gestação aceitável. Foi iniciada a validação de um instrumento comercial de medição automatizada de progesterona (eProCheck®) desenvolvido para facilitar o maneio reprodutivo. A amostra incluiu 68 fêmeas, sem diferenças significativas na proporção de raças e categorias (Vermelha Sueca, Holstein Sueca; novilha, vaca). Os animais foram incluídos no estudo após a detecção de cio e decisão da inseminação artificial (IA) pelo pessoal da vacaria. A intensidade de cio (OI) foi avaliada em paralelo pelo pessoal (NCV_OI) e pelo autor (OBS_OI). No dia 0, definido pela IA, o nível relativo de progesterona foi determinado no sangue (e leite–vacas). No dia 7, foram avaliados a distância de fuga (na manjedoura e dentro do estábulo), condição corporal, grau de claudicação e o nível de progesterona. No dia 20, o nível de progesterona foi determinado, assim como a gestação através de ultrasonografia transrectal. A gestação foi confirmada por palpação trans-rectal cerca do dia 50. Os sistemas de classificação NCV_OI e OBS_OI obtiveram resultados diferentes, tendo o segundo evidenciado OI mais elevada nas novilhas que nas vacas. A taxa de gestação global (55%) foi aceitável, tendo as novilhas maior performance que as vacas (70% vs. 37%), pelo que maior OI, em NCV_OI e OBS_OI, reflectiu-se em maior taxa de gestação. O comportamento de se deixar montar (alta intensidade de cio) correspondeu a 3.8 vezes maior probabilidade de gestação, em comparação com a detecção baseada em sinais secundários. No entanto, as IAs com base em sinais secundários, obtiveram taxa de gestação aceitável (45% baixa vs. 46% média intensidade de cio). Cios intensos (NCV_OI) corresponderam a 5.3 vezes maior probabilidade de gestação que o conjunto dos cios nítidos e fracos. Animais não-claudicantes mostraram 4.8 vezes maior probabilidade de gestação do que animais com claudicação. As distâncias de fuga foram curtas (<1m), como reflexo de uma boa interacção homemanimal na vacaria. Embora a intensidade de cio tenha tido tendência a correlacionar-se negativamente com a distância de fuga na manjedoura em vacas primíparas, as relações entre intensidade de cio e as distâncias de fuga foram consideradas inconclusivas, não sendo estatisticamente significativas. Os resultados dos ensaios preliminares do eProCheck® parecem promissores quanto ao diagnóstico precoce de gestação, devido à alta sensibilidade com um único teste ao dia 20. Ainda que seja necessário continuar a testagem, pode ser uma ferramenta complementar útil no maneio reprodutivo. Finalmente, a taxa de gestação encontrou-se associada à intensidade de cio e ao grau de claudicação, confirmando-se a importância destes factores em explorações de alta produção, embora o bem-estar animal tenha sido considerado aceitável ao nível dos parâmetros estudados. No entanto, é necessária uma reavaliação do programa de prevenção de claudicação, a fim de reduzir a sua alta prevalência nas vacas prímiparas.

An investigation of 5186 hock maps, from 3691 cows with hair loss on the hocks from 76 UK dairy farms, identified six risk factors associated with area of hair loss: higher locomotion score; poor cleanliness; higher mean milk yield; low body condition score, prolonged winter housing and certain combinations of stall base and bedding materials. Another finding was significantly poor correlation between a categorical scale and the continuous measure of hair loss. Hair loss on the hocks of 70 heifers (three herds) was observed monthly from Sep 2008-Mar 2010. The results imply that lameness precedes hair loss on the hock and not vice versa, i.e. lame animals develop hair loss rather than hock hair loss leads to future lameness. Poor cleanliness score and higher milk yield were associated positively with the risk of having hair loss on the hock. Based on the total 6889 observations from 731 cows in four herds, cows with a greater decrease in BCS (compared to BCS at calving) had higher probability of becoming lame and lower probability of recovering. Also, increase in BCS from calving was associated with lower probability of cows moving from non-lame to lame state and higher probability from lame to non-lame. Days of lactation, months of calving and parity impacted upon both non-lame to lame and lame to non-lame transitions. Analysis of 678 cows from four herds found that cows with chronic lameness had a lower probability of conception and a longer calving to conception period compared with healthy cows. Further, cows with lower average BCS had a lower probability of conception and were more likely to be lame. However, these factors didn’t influence the likelihood of an individual AI leading to pregnancy.

Master of Science
Department of Animal Sciences and Industry
Jeffrey S. Stevenson
The objectives were to evaluate the effects of equine chorionic gonadotropin (eCG) administration on preovulatory follicle diameter, serum estradiol and progesterone concentration, corpus luteum (CL) diameter, estrual activity, and pregnancy rate. Lactating dairy cows were submitted to a Presynch-Ovsynch timed artificial insemination (TAI) protocol. Cows (n = 121) in a single herd were treated with 2 injections of prostaglandin F2α (PGF) 14 d apart (Presynch), with the second injection administered 11 d before the onset of a timed AI protocol (Ovsynch; injection of GnRH 7 d before and 56 h after PGF2α, with TAI administered 16 to 18 h after the second GnRH injection). Cows were assigned randomly to receive either saline or 400 IU eCG concurrent with the PGF2α injection of the Ovsynch protocol (d 0). Blood samples were collected during the study to monitor serum changes in progesterone and estradiol to determine if eCG would facilitate increased estrual activity, improved ovulatory response to GnRH, and enhanced post-ovulatory luteal function. Administration of eCG tended to increase the number of CL and on d 9 and 16 after PGF2α, corresponding to d 6 and 13 post-ovulation. Volume of the post-eCG treatment luteal tissue was increased only on d 16. Timed AI pregnancy rates did not differ between eCG (36.9%) and control cows (41.8%). We concluded that use of eCG provided no profertility advantages to dairy cattle when programmed for a timed insemination at first service.

Contemporary reports showing negative temporal trends in human sperm quality have provided a weight of evidence, in spite of recognised methodological weaknesses of the early research, strongly indicative of declining male fertility over the past six decades. Other aspects of human male reproductive development, such as hypospadias, cryptorchidism and testis cancer are also showing negative trends in incidence and geographic variation in prevalence. Together these negative changes in parameters of fertility, which have comorbidity, are termed Testicular Dysgenesis Syndrome (TDS). The relative short time period of the trend data and the spatial heterogeneity of these factors suggest an environmental aetiology. Two neighboring and ethno-linguistic similar countries, Denmark and Finland, are used as evidence of this as Denmark has significantly decreased sperm quality as well as an increased incidence of other aspects of TDS when compared to neighboring Finland. Additionally, exposure to persistent environmental chemical pollutants has been associated with decreased sperm quality and increases in other components of TDS. The suggested underlying cause of TDS is perturbed foetal/embryonal programming and reproductive development. In spite of overall poor epidemiological information and a paucity of empirical research data from companion animals there are suggestions that environmental pollutants are affecting the health of cats and dogs. Indeed, the dog may be a relevant research model as it shares our environment and ill health associated with it, such as human TDS. Based on the evidence of environmental pollutants affecting human and animal reproductive development and fertility, this PhD project was developed in order to determine if man’s closest companion, and the animal which wholly shares our environment, demonstrates any parallel reproductive impairment. Specifically, this thesis aims to investigate negative temporal trends related to canine fertility, determine the presence of and geographic variation in tissue concentrations of exemplar environmental pollutants and examine testes tissue and sperm cells, via culture and histology, for association between chemical concentrations and quantitative assessments. Canine testes were collected from three countries; UK (n = 58), Finland (Helsinki region) (n = 20), Denmark (Copenhagen region) (n = 10). These samples were tested for concentrations of three representative chemical groups: PCBs, PBDEs, DEHP. Samples of commercial dog food (n = 28), and a range of canine body fluid was also analysed for these same chemical groups. A retrospective analysis of database information from a population of service dogs in the UK was used to determine temporal trends in canine sperm quality and cryptorchidism over a 26 year period. The concentrations of the three chemical pollutant types were additionally analysed against land use with a Geographic Information System (GIS) and two CORINE land use categories: Artificial and Agricultural. These tissues were assessed histologically using histomorphometric scores, CYP11A1 area stained. Canine testes tissue and sperm were cultured with PCB153 and DEHP to determine direct effects of short term exposures to environmental pollutants by assessing testosterone production (testes explant culture), motility (CASA), sperm viability (hypo osmotic swelling test, chromatin integrity, live:dead ratio). Database analysis of service dog fertility reported two components of TDS: negative temporal trends in sperm quality and a concurrent increased incidence of cryptorchidism. Samples of canine testes tissue contained measurable concentrations of three exemplar pollutants (PCBs, PBDEs, DEHP). These pollutants were also largely present in commercial dog food and some were also present in prostatic fluid, full ejaculate and bitches milk. The profile of EDs and concentrations of specific chemicals showed significant variation across three regions of the UK (Southeast, East Midlands, and West Midlands). Notably, concentrations of legacy industrial chemicals (PCBs, PBDEs) were highest in the West Midlands and phthalate (DEHP) was highest in the southeast. The concentrations of DEHP was significantly higher in Artificial land categories compared to tissue collected in Agricultural land use areas (P < 0.05). Canine testis samples collected from three international regions contained concentrations of PCBs and DEHP which were significantly higher in the UK (P < 0.05) and PBDEs which were significantly higher in Finland (P < 0.05). The histomorphometric assessment of testes showed significant variations across the three countries. The quantification of immunohistochemical testis area stained for CYP11A1 from the three countries showed a significant variation (P=0.0001), and tissue from Denmark showed a significant association between area stained and concentrations of DEHP (P < 0.05). Tissue exposed, via testes explant culture, to PCB153 and DEHP at two concentrations showed no significant blunting of LH stimulated testosterone secretion. Sperm exposed, via culturing with two environmentally relevant concentrations of PCB153 and DEHP, exhibited significantly increased (P < 0.05) and decreased (P < 0.05) measures of motility with these chemicals respectively. Additionally sperm viability, measured using the hypo osmotic swelling assay, was significantly reduced in the presence of PCB153 (P < 0.05) and a significant increased when cultured with DEHP (P < 0.01). Both chemicals significantly increased sperm DNA fragmentation. Additionally, short term exposure to both chemicals significantly reduced the Live:Dead ratio of sperm (P < 0.05). Robust, determinative evidence of negative trends in canine sperm quality and incidence of cryptorchidism indicate that canine and human male fertility are decreasing in parallel. An environmental aetiology is supported by the presence of environmental chemicals in canine diet, testes tissue, prostatic fluid and full canine ejaculate. Geographic differences (National and International) in testis chemical profiles and concentrations may equate with reported regional differences in human TDS. In support of this hypothesis, specific canine testis chemical concentrations were associated with histomorphometric scores from the same cohort of samples. Direct effects of PCB153 and DEHP on sperm motility and viability may be indicative of an alternative acute negative effect on male fertility. These data suggest that the dog, while living in a shared environment with humans, exhibits some parameters of TDS and that canine, as well human, male fertility is declining due to exposure to environmental chemicals.

As nutrition is of central importance to cattle fertility, this study sought to assess how veterinarians and nutritional advisers manage trace element imbalance in the UK; diagnosis and treatment. The study also sought to develop a robust system for oocyte recovery (ovum pick-up (OPU)) and in vitro embryo production (IVP) for commercial use, and to identify key factors influencing success, including oestrus synchrony and ovarian stimulation prior to OPU. The intention originally was to use OPU/IVP to investigate the impact of mineral imbalances on bovine oocyte quality, early embryo development and pregnancy establishment following embryo transfer (ET). In the first survey of its kind in the UK, the understanding and approach of advisers to mineral nutrition on farms was investigated. Of the 173 respondents, 78% were vets in practice. The overall importance of minerals was recorded by vets as low 33%, medium 37%, and high 30%, while non-vets scored importance as low 17%, medium 48%, and high 35%. There was little consensus amongst the advisers, or within the vet and non-vet subgroups about mechanisms and interactions associated with deficiency, and particularly of copper responsive conditions. The most frequently identified deficiencies were selenium, copper and iodine, while the most commonly identified toxicity was molybdenum. For copper responsive conditions, all of the listed treatments were used at least "occasionally"; the most frequently being glass boluses, in-feed supplementation, matrix boluses, and then copper injections. While there was a diverse choice of treatments, altering the ration was relatively rarely selected. This thesis also provides the first large-scale retrospective analysis of factors influencing the establishment of a commercially robust ovum pick up (OPU) and in vitro embryo production (IVP) platform in the UK. Over a 5-year period, a system was developed and validated for use in the UK with 2,138 cycles of OPU. These cycles were analysed as four sets of data and included two IVP laboratories and 6 OPU teams. Factors in these analyses included OPU team, IVP laboratory, ovarian stimulation protocol and semen type (unsorted vs sex-sorted). The mean number of follicles aspirated by the OPU teams ranged from 6.5 to 14.9 (P < 0.001), while the number of oocytes collected was between 4.0 and 12.4 (P < 0.001). There was an indication (P=0.055) that the blastocyst per oocyte rate varied between teams. The proportion of blastocysts from oocytes that cleaved was higher (P=0.01) for unsorted than sexed semen. Two commercial products containing different ratios of follicle stimulating hormone (FSH) to luteinising hormone (LH) (Folltropin® and Pluset®) were compared in ovarian stimulation programs. The addition of 'coasting' (short-term (typically 48h) hormonal withdrawal after FSH stimulation), prior to OPU was also investigated. Pluset® resulted in a greater (P < 0.001) mean number of follicles aspirated, more (P=0.003) blastocysts per oocyte matured and more (P < 0.001) embryos per cycle (2.45), compared with Folltropin® (1.17) or with no stimulation (1.24). Throughout the study there was a steady improvement in blastocyst production per OPU cycle. In a separate analysis, Grade 1 cumulus oocyte complexes (COCs) as a proportion of COCs recovered, oocytes that cleaved as a proportion of total COCs, and blastocysts as a proportion of total COCs, were all greater (P < 0.05) for stimulated than non-stimulated cycles, irrespective of FSH/LH product. A composite score of oocyte quality and quantity was proposed (sCOC); Log Total Mean sCOC was correlated (P < 0.001) with both the proportion of blastocysts per oocyte collected, and the total number of embryos produced per cycle. Finally, twelve peri-pubertal heifers (approximately 10 months old) participated in a crossover trial which compared PRID® (Delta®) vs CIDR® progesterone releasing intravaginal devices for use in OPU/IVP cycles. Vaginoscopic examination found higher vaginal inflammation grades for PRID® than CIDR® (P < 0.001). There was evidence of vaginal inflammation continuing for at least 2 weeks after device withdrawal. The proportion cleaved of oocytes inseminated was higher for PRID® than CIDR® (P < 0.05). Numerically but not significantly there was a higher proportion of blastocysts per cycle and a higher Log Total Mean sCOC score per cycle with PRID® than CIDR® treatments, but blastocyst yield was low throughout, suggesting a need to repeat the trial. Data collection and analyses are ongoing, to identify other key performance indicators within the OPU/IVP embryo transfer (ET) system, with a view to refining the sCOC composite score model. A robust OPU/IVP/ET system has been developed and this could be used to investigate further how mineral imbalances impact oocyte competence and blastocyst yield.

Chromosome evaluations were made on leukocyte blood samples from 169 phenotypically normal nulliparous Holstein heifers. These were from three different reproduction groups collected in sets of threes from 10 different herds in the western United States. Group 1, the control group, consisted of heifers diagnosed pregnant after one or two breedings; Group 2 consisted of heifers diagnosed pregnant after three or four breedings; and Group 3 consisted of heifers diagnosed open after four or more breedings. Metaphase chromosome spreads used in these analyses were obtained through 72-hour leukocyte cultures from heparinized whole blood. Processed cells were dropped onto a slide, air-dried, and stained with Giemsa. Chromosomes were then counted and X chromosomes were identified. A total of 1, 597 cells was evaluated, with 1,439 cells having counts of 60 chromosomes each. Thirty cells had less than 58 chromosomes, 31 cells had 58 chromosomes, 75 cells had 59 chromosomes, 14 cells had 61 chromosomes, and 8 cells had more than 61 chromosomes (9 .98% were different than 60). All counts of other than 60 chromosomes were isolated cases and were not identified as abnormalities. Only two cells from two separate heifers contained what may have been sex chromosome abnormalities. No persistent chromosomal defects were observed among the 169 heifers.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
O uso generalizado da inseminação artificial (IA) contribuiu grandemente para o sucesso da indústria de suínos, por meio do auxílio e disseminação do progresso genético. Atualmente, reprodutores suínos jovens são selecionados para IA com base em seus valores genéticos para características de produção e, a seleção de reprodutores para características de sêmen, como volume, concentração, motilidade e morfologia, bem como para menor variação intra- reprodutor na sua produção e qualidade, ainda não é uma prática comum. Esta seleção é importante para melhorar o desempenho dos reprodutores nas estações de IA, cujo objetivo é maximizar o número de doses inseminantes produzidas por cada ejaculado. A estimação de parâmetros genéticos e quantificação da variação genética para características de sêmen e para variação intra-reprodutor permitem analisar se essas características devem ser incluídas nos objetivos do melhoramento. Além da estimação de parâmetros genéticos para fins de seleção, o interesse em estudar os processos moleculares e os mecanismos genéticos que afetam as características de sêmen está aumentando nos últimos anos. Os estudos de associação genômica ampla (GWAS) são comumente usados para identificar polimorfismos de base única (SNPs) associados a loci de características quantitativas (QTL) com maiores efeitos. O GWAS em passo único ponderado (WssGWAS) é um método que permite a estimação de efeitos de SNP utilizando informações de todos os animais genotipados, fenotipados e com pedigree na população. Expandindo as fronteiras dos estudos de reprodução em suínos, outro campo importante a ser explorado em programas de melhoramento é a fertilidade dos reprodutores. As características reprodutivas, como a duração da gestação (GL), o número total de leitões nascidos (TNB) e nascidos mortos (SB) são algumas características-chave para a produção eficiente de suínos. Devido às baixas ou moderadas herdabilidades para essas características, é importante identificar todos os fatores que as influenciam e incluir esses fatores nos modelos de avaliação genética. Os efeitos do reprodutor cujo ejaculado foi utilizado para inseminar a matriz e do ejaculado são dois desses fatores importantes que têm o potencial de melhorar os modelos tradicionais utilizados nas avaliações genéticas das características reprodutivas. Dentre os elementos que controlam o tamanho da leitegada, as taxas de fertilização e de sobrevivência pré-natal podem ser influenciadas pelo reprodutor, devido às diferenças genéticas na capacidade de fertilização relacionadas à qualidade do sêmen e/ou à contribuição genética do reprodutor para a viabilidade do embrião. Nesse contexto, os objetivos gerais com este estudo foram 1) estimar os parâmetros genéticos para qualidade e quantidade de sêmen, bem como para a variação intra-reprodutor para essas características; 2) identificar regiões de QTL e genes candidatos associados a características de sêmen por meio do WssGWAS e, subsequentemente, realizar análises de redes gênicas para investigar os processos biológicos compartilhados por genes identificados em diferentes linhas de suínos e 3) estimar parâmetros genéticos para o efeito do reprodutor na GL, TNB e SB e avaliar a inclusão dos efeitos do reprodutor e do ejaculado nos modelos de avaliação genética dessas características. Os resultados desta tese mostraram estimativas moderadas de herdabilidade e correlações genéticas favoráveis entre características de sêmen, indicando que a seleção de reprodutores para essas características pode resultar em razoável progresso genético. Além disso, variação genética relevante foi encontrada para a variabilidade intra-reprodutor para essas características, revelando a possibilidade de seleção de reprodutores para uma menor variação na qualidade e produção de sêmen. Os resultados do WssGWAS apontaram regiões relevantes de QTL que explicaram grandes proporções da variância genética (até 10,8%) para as características de sêmen em vários cromossomos suínos, confirmando a suposição de complexidade genética dessas características. Esta identificação foi possível com o baixo número de animais com fenótipos e genótipos, devido à escolha apropriada do método. Os genes candidatos SCN8A, PTGS2, PLA2G4A, DNAI2, IQCG, LOC102167830, NME5, AZIN2, SPATA7, METTL3 e HPGDS foram identificados associados às características de sêmen nas regiões de QTL identificadas para as linhas de suínos avaliadas. A análise de redes gênicas mostrou genes candidatos encontrados para diferentes linhas de suínos compartilhando caminhos biológicos que ocorrem nos testículos de mamíferos. No que diz respeito à fertilidade do reprodutor, os resultados mostraram que há variação genética devido ao efeito do reprodutor em GL, TNB e SB; e o modelo com inclusão de efeitos de ambiente permanente e genéticos do reprodutor, além do efeito do ejaculado, mostrou o melhor ajuste para os dados. Esta tese resultou em informações científicas importantes e inovadoras na área de reprodução em machos, o que contribuirá para aumentar o conhecimento ainda escasso sobre a seleção genética e a arquitetura genômica de características de qualidade de sêmen e de fertilidade em reprodutores suínos.
The widespread use of artificial insemination (AI) has greatly contributed to the success of the pig industry by assisting and disseminating the genetic progress. Currently, young boars are selected for AI based on their breeding values for production traits and selecting boars for semen traits, such as volume, concentration, motility and morphology, and for low variation in semen quality and production is still not a common practice. This selection is important for better performance of boars at AI stations, whose objective is to maximize the number of insemination doses produced by each ejaculate. The estimation of genetic parameters and the quantification of genetic variation for semen traits and within-boar variation allow an analysis of whether these traits should be included in the breeding goal. Besides the estimation of genetic parameters for selection purposes, the interest in studying the molecular processes and genetic mechanisms affecting semen traits is increasing in recent years. Genome-wide association studies (GWAS) are commonly used to identify single nucleotide polymorphisms (SNPs) associated with quantitative trait loci (QTL) with major effect. The weighted single-step GWAS (WssGWAS) is a method that allows estimation of SNP effects using information from all genotyped, phenotyped and pedigree animals. Expanding the frontiers of reproduction studies in pigs, another important field to be explored in breeding programs is the boar fertility. Reproductive traits, such as gestation length (GL), total number of piglets born (TNB) and stillborn (SB) are some of the bottleneck traits for efficient pig production. Because of the low to moderate heritabilities for these traits, it is important to identify all factors influencing them and to include these factors in the genetic evaluation models. The service sire (boar which ejaculate dose was used to inseminate the sow) and ejaculate effects are two of those important factors that have the potential to improve the traditional models used in the genetic evaluations of reproductive traits. Among the elements controlling the litter size, the fertilization rate and prenatal survival rate might be influenced by the service sire due to genetic differences in the capacity of fertilization, which is related to sperm quality and/or the boar genetic contribution to viability of the embryo. In this context, my overall aims were 1) to estimate genetic parameters for semen quality and quantity traits, as well as for within-boar variation of these traits; 2) to identify QTL regions and candidate genes associated with semen traits through a WssGWAS and, subsequently, to perform gene network analyses to investigate the biological processes shared by genes identified in different pig lines and 3) to estimate genetic parameters for service sire on reproductive traits GL, TNB and SB and to evaluate the inclusion of service sire and ejaculate effects in the genetic evaluation models of these traits. The results of this thesis showed moderate estimates of heritability and favorable genetic correlations between semen traits, indicating that boar selection for these traits could make reasonable genetic progress. In addition, relevant genetic variation was found for within-boar variability of these traits, revealing the possibility of selection of boars for reduced variation in semen quality and production. Results from WssGWAS pinpointed relevant QTL regions explaining high proportions of genetic variance (up to 10.8%) for semen traits in several pig chromosomes, confirming the assumption of genetic complexity of these traits. This identification was possible with low number of animals having both phenotypes and genotypes due to the appropriate choice of the method. Candidate genes SCN8A, PTGS2, PLA2G4A, DNAI2, IQCG, LOC102167830, NME5, AZIN2, SPATA7, METTL3 and HPGDS were identified associated with semen traits in the QTL regions identified for the pig lines evaluated. The gene network analysis showed candidate genes found for different pig lines sharing biological pathways that occur in mammalian testes. Regarding boar fertility, the results showed that there is genetic variation due to service sire effect on GL, TNB and SB; and the model with inclusion of permanent environmental and genetic effects due to service sire, in addition to ejaculate effect, showed the best fit to the data. This thesis resulted in important and innovative scientific information on male reproduction field in pigs, which will contribute to increase the still scarce knowledge about genetic selection and genomic architecture of boar semen quality and fertility traits.

Actuellement en France et aux Etats-Unis, seules 30 à 35 % des premières inséminations artificielles chez la vache Holstein aboutissent à un vêlage. Les échecs de gestation ont lieu le plus souvent au cours du développement embryonnaire (Non-Fécondation-Mortalité Embryonnaire Précoce (NF-MEP) entre 0 et 16 jours, Mortalité Embryonnaire Tardive (MET) entre 16 et 45 jours après insémination). Ces échecs précoces de gestation expliquent en grande partie les différences de fertilité observées en France entre la race Holstein et les autres races plus fertiles. Les facteurs de risque de l'interruption de la gestation sont multiples : liés à l'animal, à l'environnement et à la conduite de l'élevage. Trois facteurs cependant ont évolué au cours du temps pour les vaches Holstein : le potentiel génétique, les anomalies de reprise de la cyclicité après vêlage, le déficit énergétique en début de lactation. Notre objectif a été d'identifier l'influence de ces trois facteurs sur les échecs précoces de gestation après première insémination postpartum chez la vache de race Holstein. Le potentiel génétique a été associé à l'incidence de NF-MEP et de MET. Les phases lutéales prolongées sembleraient être à l'origine d'une augmentation d'incidence de MET. Le déficit énergétique a eu un effet sur la NF-MEP et sur la MET. L'effet du déficit énergétique postpartum sur la qualité des ovocytes au moment de la mise à la reproduction reste hypothétique. Notre travail sur les échecs précoces de gestation a permis de contribuer à fournir des outils d'amélioration de conduite d'élevage aux professionnels.

Commonly used soil analysis and resin capsule procedures are used to assess nutrient status in fertile soils, but their validity in semi-arid ecosystems is unknown. Three studies were performed to assess resin capsule effectiveness in semi-arid ecosystems. An incubation study was completed in which loamy sand and sandy clay loam soils were treated with rates of N, P, Fe and Zn. Each soil treatment was implanted with a resin capsule and incubated for 60 or 120 days. Resin capsules reflected NH4-N and P fertilizer at low rates in the loamy sand. NO3-N reflected rates in both soils, but did not reflect Fe or Zn application. Resin capsule NH4-N was a better indicator than KCl-extractable NH4-N, but resin capsule NO3-N was not as effective as water extraction, and resin capsule P was poor compared to NaHCO3-P. A second study was performed in glasshouse conditions using the incubation study soils. Soils were treated with rates of N, P and resin capsules were placed in pots. Pots were seeded with squirreltail grass (Elymus elymoides) and placed in a glasshouse. Resin capsules were removed at 120 days, soil samples taken, grass harvested and yield measured. Yield and total nutrient removal was correlated to resin NH4-N, marginally related to resin or soil NO3-N, and unrelated to resin P. Yield and total nutrient removal was correlated with application rates and resin NH4-N and NaHCO3-extracted P. The third field study, compared two sites with rates of N and P application were established on clay loam and sandy loam soils. Resin capsule and conventional soil tests for NO3-N, NH4-N and P were measured and plant nutrient status examined. Resin capsules were removed and replaced and soil samples taken every 90 days. Resins P was not related to P application or to plant tissue P but NaHCO3-extracted P was, while resin NO3-N, KCl-extracted NO3-N and NH4-N were correlated to N application and plant N. Soil test P was more effective in predicting P status and bioavailability than resin capsules. Resin NH4-N and NO3-N predicted N status and bioavailability, but soil tests were just as effective in semi-arid conditions.

Current models for predicting the environmental fate of munitions constituents (MC) in soils are based mostly on chemical distribution parameters and neglect the larger considerations of other soil parameters. We are working towards a new approach based on the whole soil activity for predicting the long-term residence time and fate of MC in soils with connection to agronomic soil fertility concepts. These relationships are demonstrated by correlating experiments involving full physical and chemical characterization of eight taxonomically distinct soils, with batch reactor studies determining MC degradation potential. Soils were incubated in a slurry for 10 days in a closed reactor system with temperature, pH, and Eh readings recorded every 24 h. Air and slurry samples were taken eight times within the 168 h incubation period. Carbon dioxide, TNT, and nutrient solution concentrations were assessed to obtain a full picture of soil chemistry changes associated with microbial activity in response to additions of TNT. Multivariate analysis was used to determine the main factors impacting degradation rate. Principle Components Analysis (PCA) statistically classified the soils based on the variance of their soil property data. Partial Least Squares Analysis (PLS) showed that TNT degradation was possibly correlated with soil fertility characteristics. Although this is an exploratory study, results show promise in moving towards a more effective way of predicting TNT environmental fate in soils.

Le gène BCAR4 (Breast Cancer anti-estrogen Resistance 4) a précédemment été découvert chez le bovin comme un gène exprimé préférentiellement dans l'ovocyte et l'embryon précoce et dont l’inhibition altère le développement embryonnaire in vitro. Cependant, son rôle dans l’ovogenèse, la folliculogenèse et globalement la fertilité in vivo restait inconnu. Le gène est conservé chez divers mammifères mais pas chez les rongeurs, le lapin a donc été choisi pour analyser son expression et sa fonction in vivo. Le transcrit BCAR4 est détecté dans l’ovaire dès la formation des follicules primordiaux, et dans les follicules préantraux et antraux, jusque dans l’ovocyte ovulé. Son abondance diminue après la fécondation et tout au long du développement préimplantatoire pour disparaître dans le blastocyste, un profil typique d’un transcrit maternel.Afin d’appréhender le rôle du gène BCAR4 in vivo dans la reproduction chez la femelle, des lapins porteurs d’une altération du gène BCAR4 ont été créés par édition du génome à l’aide de nucléases effectrices de type activateur de transcription (TALEN), et une lignée a été générée. Les animaux porteurs de la modification à l’état homozygote ou hétérozygote sont viables et en bonne santé. L’efficacité de l’altération génétique a été démontrée par transcription inverse couplée à la PCR : trente fois moindre que chez les animaux de génotype sauvage, l’expression de BCAR4 peut être considérée comme abolie dans les follicules ovariens des individus homozygotes. Les femelles ont été phénotypées sur plusieurs paramètres liés à la reproduction. Le génotype n’a pas d’impact significatif sur la folliculogenèse ou l’activité ovarienne, estimés par le dénombrement folliculaire sur coupes histologiques, la concentration plasmatique en hormone anti-mullérienne ou la réponse à la stimulation ovarienne. Pour évaluer la fertilité et la prolificité, les femelles ont été inséminées trois fois à six semaines d’intervalle. Les femelles homozygotes présentaient un taux de mise-bas significativement plus faible que les femelles hétérozygotes, 22±7% vs 71±11% (moyenne±sem), et une prolificité de 1,5±0,7 vs 5,8±1,5 lapereaux par insémination. En conclusion, le gène BCAR4 n’est pas indispensable pour le développement folliculaire, mais il contribue à une fertilité optimale chez la lapine
The BCAR4 gene (Breast Cancer anti-estrogen Resistance 4) has been previously characterized in cattle as a gene expressed preferentially in the oocyte and early embryo, whose inhibition alters embryonic development in vitro. However, its role in oogenesis, folliculogenesis and overall in fertility in vivo remains unknown. Since this gene is conserved in various mammals but not in rodents, the rabbit has been chosen to investigate its expression and function in vivo. By reverse transcription coupled to PCR, BCAR4 transcript is detected in the ovary when primordial follicles are formed, and in ovarian follicles at the preantral and antral stages, as well as in ovulated oocytes. Its abundance decreases after fertilization and throughout preimplantation development to disappear in the blastocyst, a typical profile for a maternal transcript.In order to elucidate the role of BCAR4 in vivo in female reproduction, rabbits carrying an altered BCAR4 gene were created and a line was generated. Both homozygous and heterozygous carriers of the genetic alteration are viable and appear healthy. The genetic alteration abolishes BCAR4 expression in ovarian follicles of homozygous animals, as the transcript abundance is down thirty-fold as compared to the wild-type phenotype. Females were phenotyped on several parameters related to reproduction. The genotype did not have a significant impact on follicular development or ovarian activity, as estimated by follicular count onto ovarian sections, anti-mullerian hormone concentration in plasma, and the response to ovarian stimulation. To evaluate their fertility and prolificacy, females were inseminated three times every six-weeks. Homozygous females had a significantly lower farrowing rate than heterozygous females, 22±7% vs 71±11% (mean±sem), while prolificacy was 1.5±0.7 vs 5.8±1.5 pups per insemination. In conclusion, BCAR4 is not essential for follicular development but the gene contributes to optimal fertility of female rabbits

An intrinsic link exists between soil moisture and soil nitrogen. Factors that increase or decrease soil moisture can have a profound effect on soil nitrogen cycling, which may have later repercussions in the plant community. Post-fire soil water repellency is one factor that can limit soil moisture acquisition and may indirectly affect nitrogen cycling and weed invasion in woody islands of fertility. Plots centered on burned Juniperus osteosperma trees were either left untreated or treated with a surfactant to ameliorate water repellency. Two years later, soils were excavated from the untreated and treated field plots. In the greenhouse, half of each soil type received a surfactant treatment while the other half was left untreated. Pots were seeded with either Bromus tectorum or Pseudoroegneria spicata. Analysis of field soil prior to the greenhouse trial showed that untreated, repellent soils had inorganic nitrogen levels an order of magnitude higher than wettable, surfactant-treated soils. Greenhouse pots that had received a surfactant treatment in the field and/or greenhouse had similar soil water content, plant density, and above ground biomass, which were, respectively, 55-101%, 31 to 34 -fold, and 16 to 18 -fold greater than pots without a surfactant treatment. No species effects were found. This study indicates that water repellency can reduce wetting and retention of water in the soil while promoting the retention of high levels of inorganic nitrogen. However, the effects of soil water repellency on inorganic nitrogen appeared to have a minimal effect on plant growth compared to the effect of soil water repellency on water availability.

Thesis (MScAgric)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: Profitable milk production and genetic improvement in dairy herds are largely dependant on fertile cows capable of calving down on an annual basis. Several studies indicate declines in the reproductive performance of Holstein cows over the last 30 years. Calving interval (CI) and services per conception (SPC) are being used by dairy farmers as indicators of the reproductive performance of dairy cows. However, using these traits as cow fertility indicators is problematic as CI is dependent on subsequent calving dates while SPC is strongly linked to inseminator proficiency. The aim of the study is therefore, firstly, to describe alternative fertility traits derived from insemination and calving dates and pregnancy check results. The effects of some non-genetic factors on these traits are discussed. Means±sd for interval traits from calving to first insemination (CFS) and the interval from calving to conception (DO) were respectively 77±30 and 134±74 days while the number of services per conception (SPC) averaged 2.55±1.79. The percentage of first servics occurring within 80 days post-partum (FS80d) and the proportion of cows being confirmed pregnant within 100 (PD100d) and 200 days post-partum (PD200d) were 0.64±0.48, 0.36±0.48 and 0.71±0.45, respectively. Although fertility traits were affected significantly by lactation number, calving year and month, herds (managers) had the largest effect. Fertility is a complex trait, however, the challenge is finding traits that best describe this trait. Genetic parameters for these traits could give an indication of the response to selection in dairy herds. In the second part of this study, genetic parameters and correlations between fertility traits, sourced from standard reproduction management data bases, are analysed for Holstein cows using bivariate linearlinear and linear-threshold animal models. Insemination events (n = 69 181) from 26 645 lactations of 9 046 Holstein cows from 14 herds, calving down during the period from 1991 to 2007, were available. The outcome of each AI event was known. Insemination records were linked to the calving date of each cow, lactation number as well as dam and sire identification. Fertility traits indicating the ability of cows to show heat early in the breeding period, and to become pregnant, were derived. Data were analysed using bivariate linear-linear and linear-threshold animal models with fixed effects being herd (14 levels), year (17 levels), season (4 levels) and lactation number (6 levels). The model included the random effects of animal and animal permanent environment (PE). Heritability estimates ranged from 0.04±0.01 to 0.10±0.02 for FS80d, from 0.07±0.01 to 0.08±0.02 for PD100d and from 0.06±0.04 to 0.08±0.02 for PD200d depending on the two-trait combination. Although heritability estimates of most fertility traits were below 0.10, they were in close agreement with results published by other researchers using linear models. Genetic correlations between different fertility parameters analyzed in this study indicated that it is unlikely that a single characteristic would serve well for selection purposes; instead, combining different traits could be used in selection programmes to improve fertility. Further research in constructing an optimal fertility index is warranted. In the third part of this study, genetic parameters for South African Holstein cows for fertility and production traits were estimated from 2415 lactation records. Two-trait analysis of fertility and milk yield was investigated as a method to estimate fertility breeding values when culling, or selection based on milk yield in early lactation, determines presence or absence of fertility observations in later lactations. Fertility traits were days from calving to first service (CFS), days from calving to conception (DO), percentage cows inseminated within 80d post-partum (FS80d), number of service per conception (SPC), and the binary traits percentage of cows pregnant within 100d and 200d postpartum (PD100d, Pd200d). Milk production traits were 300 day milk, fat and protein yield. For fertility traits, range of estimates of heritability (h2) was 0.006 to 0.08 for linear traits and 0.05 to 0.12 for binary traits. The range for permanent environmental variance (c2) was 0.016 to 0.032. In this study genetic correlations of fertility with milk production traits were unfavourable ranging between −0.93 to 0.76.
AFRIKAANSE OPSOMMING: Winsgewende melkproduksie en genetiese verbetering in melkkuddes hang grootliks af van vrugbare koeie wat op ‘n jaarlikse basis kalf. Verskeie studies toon dat die reproduksievermoë van Holsteinkoeie oor die afgelope 30 jaar afgeneem het. Melkboere gebruik tussenkalfperiode (TKP) en aantal inseminasies per konsepsie (KIPK) as aanduidings van die reproduksievermoë van melkkoeie. Dit is egter moeilik om hierdie eienskappe as vrugbaarheidseienskappe vir melkkoeie te gebruik omdat TKP afhanklik is van opeenvolgende kalfdatums terwyl KIPK baie sterk gekoppel is aan die inseminasievermoë van die insemineerders. In die eerste gedeelte van die studie word alternatiewe vrugbaarheidseienskappe, wat afgelei is van inseminasie- en kalfdatums en die uitslag van dragtigheidsondersoeke, beskryf. Die invloed van ‘n aantal nie-genetiese faktore op dié eienskappe word ook bespreek. Gemiddeldes±standaard afwykings vir die periode vanaf kalwing tot eerste inseminasie (CFS), die periode van kalwing tot konsepsie (DO) was 77±30 en 134±74 dae onderskeidelik, terwyl die aantal inseminasies per konsepsie (SPC) 2.55±1.79 was. Die persentasie eerste inseminasies wat binne 80 dae na kalwing (FS80d), en die persentasie koeie wat dragtig bevestig is binne 100 (PD100d) en 200 dae na kalwing (PD200d) was 0.64±0.48, 0.36±0.48 en 0.71±0.45, onderskeidelik. Hoewel vrugbaarheidseienskappe betekenisvol beïnvloed is deur laktasienommer, jaar en seisoen van kalwing, het kuddes (bestuurders) die grootste effek op eienskappe gehad. Vrugbaarheid is ‘n ingewikkelde eienskap en die uitdaging is om eienskappe te vind wat dit die beste beskryf. Genetiese parameters vir eienskappe wat oorweeg word sal ‘n aanduiding gee van die seleksieresponse in melkkuddes. In die tweede gedeelte van die studie is genetiese parameters van vrugbaarheidseienskappe en korrelasies tussen dié eienskappe beraam. Eienskappe is bekom vanaf reproduksiebestuursprogramme wat in melkkuddes gebruik word. Al die inseminasierekords (n = 69 181) van 26 645 laktasies van 9 046 Holsteinkoeie van 14 melkkuddes wat tussen 1991 en 2007 gekalf het, was beskikbaar. Die uitslag van elke inseminasie was bekend. Inseminasierekords is met die kalfdatum, laktasienommer, identifkasienommers van die moeder en vader van elke koei, gekoppel. Vrugbaarheidseienskappe wat die vermoë van koeie aandui om vroeg na kalwing op hitte te kom en beset te raak, is verkry. Die data is ontleed deur gebruik te maak van twee-eienskap liniêreliniêr- en liniêr-drempel-diere modelle met vaste effekte kudde (14 vlakke), jaar (17 vlakke), siesoen (4 vlakke) en laktasienommer (6 vlakke). Die modelle het die ewekansige effekte van dier- en dierpermananente omgewingseffkte (PE) ingesluit. Genetiese, diere PE en residuele korrelasies is vervolgens beraam. Oorerflikhede varieer vanaf 0.04±0.01 tot 0.10±0.02 vir FS80d, vanaf 0.07±0.01 tot 0.08±0.02 vir PD100d en vanaf 0.06±0.04 tot 0.08±0.02 vir PD200d, afhangende van die tweeeienskap kombinasie. Ten spyte daarvan dat die oorerflikhede van die meeste vrugbaarseienskappe laer as 0.10 is, is die resultate in ooreenstemming met ander navorsers wat liniëre modelle gebruik het. Genetiese korrelasies tussen verskillende vrugbaarheidseienskappe toon dat daar nie enkel beste eienskap is wat vir seleksiedoeleindes gebruik kan word nie. Dit sou waarskynlik beter wees om verskillende eienskappe te kombineer om die vrugbaarheid in melkkoeie te verbeter. Verdere navorsing is nodig om ‘n optimale vrugbaarheidseienskap te ontwikkel. Dit is belangrik dat die verband tussen reproduksie en melkproduksie vir melkkoeie bepaal word. In die derde gedeelte van die studie is genetiese parameters vir vrugbaarheid- en melkproduksieeienskappe vir Suid Afrikaanse Holsteinkoeie beraam. ‘n Totaal van 2415 laktasierekords was beskikbaar. Vrugbaarheid en melkproduksie is volgens ‘n twee-eienskap analise ontleed as ‘n metode om teelwaardes vir vrugbaarheid te bepaal in gevalle waar die uitskot of seleksie gebaseer op melkproduksie in vroeglaktasie die teenwoordigheid of afwesigheid van vrugbaarheidseienskappe in latere laktasies bepaal. Vrugbaarheidseienskappe was die periode (aantal dae) tussen kalfdatum en eerste inseminasie (CFS), die aantal dae van kalf tot konsepsie (DO), die persentasie koeie wat by 80 dae na kalf vir die eerste keer geïnsemineer was (FS80d), die aantal inseminasies per konsepsie (SPC), en binêre eienskappe van die persentasie koeie wat by 100 dae en 200 dae na kalf beset was (PD100d en PD200d). Melkproduksie-eienskappe was 300-dae melk., vet- en proteïnproduksie. Vir vrugbaarheidseienskappe het die oorerflikheidswaardes (h2) vanaf 0.006 tot 0.08 vir liniêre eienskappe gevarieer en tussen 0.05 tot 0.12 vir binêre eienskappe. Die permanente omgewingsvariansie (c2) het tussen 0.016 tot 0.032 gevarieer. In hierdie studie was die genetiese korrelasies tussen vrugbaarheidseienskappe en melkproduksie-eienskappe ongunstig en het dit tussen −0.93 tot 0.76 gevarieer.

Equine spermadhesin HSP-7 is a 14 kDa protein isolated from stallion seminal plasma and present on the surface of spermatozoa. HSP-7 displays carbohydrate and zona-pellucida binding properties, but the physiological role in equine fertilization is not well defined. HSP-7 has 98% amino acid sequence homology with the well-studied boar spermadhesin, AWN. Currently, these two proteins are considered to have the same reproductive function. Immunofluorescence studies presented here show that the stallion and boar spermadhesins are localized to different segments on spermatozoa. The variation in molecular compartmentalization of spermadhesin molecules in different species suggests that these structurally related proteins could be involved in independent events of fertilization. While the variation in HSP-7 abundance was not statistically significant between fertile and subfertile stallions, capacitated spermatozoa displayed a marked increase in HSP-7 when compared to neat sperm (P < 0.05). These results indicate that rather than aiding in capacitation, HSP-7 is exposed with capacitation and may have a more significant role in the acrosome reaction and sperm-oocyte recognition than previously documented.

Dans la nature, les relations symbiotiques sont très répandues, et d’une importance écologique fondamentale. Les animaux sont apparus, ont évolué et vivent maintenant constamment associés avec une multitude de micro-organismes. Parmi les différents types de symbioses existantes, celles liant le microbiote et son hôte occupent une place centrale et équilibrée, basée sur des relations commensales ou mutualistes entre les partenaires. Ce microbiote est de plus en plus étudié, notamment en raison du rôle crucial qu’il joue dans la santé animale ainsi que dans le développement de pathologies. Dans cette effort de recherche, Drosophila melanogaster représente un modèle de choix, grâce à la facilité de générer et maintenir des lignées de mouches axéniques, ainsi que de les réassocier avec une communauté microbienne définie.L’association de la drosophile avec l’un des ses commensaux naturels, Lactobacillus plantarum, a permis de révéler l’effet promoteur de croissance de cette bactérie. En cas de carence nutritionnelle, des larves associées avec L. plantarum se développent beaucoup plus rapidement que leurs semblables axéniques. L’ajustement du développement en fonction des conditions environnementales est cependant crucial pour la formation d’un individu à la santé optimale, et dans ce cas les individus grandissent plus vite alors que les conditions nutritionnelles sont pauvres. Nous avons donc cherché à déterminer si ce qui semble être un avantage au stade larvaire pouvait se révéler délétère pour les stades suivants et avoir un effet néfaste sur les mouches adultes. Nous avons montré que L. plantarum est bénéfique pour D. melanogaster tout au long du cycle de vie de la mouche et permet l’émergence précoce d’adultes matures et fertiles sans impact négatif sur leur santé et leurs performances. De plus, dans certaines conditions, cette souche commensale entraîne une augmentation de la durée de vie de mâles nutritionnellement carencés.Des études plus larges analysant l’interaction de la drosophile avec plusieurs espèces bactériennes peuvent informer sur la dynamique d’un microbiote de mouche. En effet, au sein de la niche environnementale, les bactéries sont échangées entre l’animal et son substrat nutritif, et ces transferts réciproques pourraient altérer la composition de la communauté. Nous avons étudié cette question en utilisant un microbiote naturel, et avons observé un haut degré de similitude entre les bactéries associées avec les mouches et la composition de la communauté bactérienne de la nourriture, illustrant le caractère stable de l’association du microbiote de la drosophile avec la population de mouches au sien de la niche.Ces résultats illustrent le pouvoir du modèle drosophile pour l’étude des interactions entre les animaux et leur microbiote, qui permet de déchiffrer la dynamique des communautés de bactéries commensales ainsi que leur impact sur la physiologie animale
In nature, symbiotic relationships are widespread, and of paramount ecological importance. Animals have appeared, evolved, and are now living constantly associated with a variety of microorganisms. In the spectrum of different symbioses types, the microbiota occupies a central and balanced part by establishing commensalistic or mutualistic relationships with its host. Over the last years, the microbiota has been extensively studied given the crucial role it plays in animal health and disease. In this research effort, Drosophila melanogaster represents a fruitful model, thanks to the ease to generate and maintain axenic flies, and the simplicity of re-associating them with a defined microbial community.The association of Drosophila with one of its natural commensals, Lactobacillus plantarum, revealed a growth-promoting effect mediated by this bacterial species. In case of nutrient scarcity, larvae associated with L. plantarum develop twice faster than the germ-free ones. However, adjusting development to environmental cues is key to organismal fitness, and yet here animals are growing fast even though the nutritional conditions are poor. We thus questioned whether what seems like an advantage could in turn be deleterious at later stages, and adversely impact adult fitness. We showed that L. plantarum is a true beneficial partner for D. melanogaster throughout the fly life cycle. Indeed, it allows the precocious emergence of mature and fertile adults without fitness drawbacks, and in certain conditions, this commensal can even increase the lifespan of nutritionally challenged males.Broader studies assessing the interaction of Drosophila with several bacterial species can inform about the dynamics of a fly microbiota. Indeed, in the environmental niche bacteria are transferred between the fly and its nutritive substrate, and these reciprocal transfers could alter the composition of the community. We addressed this question using a wild-derived microbial community and observed a high degree of similarity between the bacteria associated with the flies and the composition of the community in the diet, illustrating the stable association of the Drosophila microbiota with the fly population in the niche.Altogether these results emphasize the power of the Drosophila model in the study of the relationships between animals and their microbiota, which allows deciphering the dynamics of commensal bacterial communities and their impact on animal physiology

This study was carried out in swine production facilities of the School of Veterinary Medicine and Zootechnic Studies at the Rural Academic Unit of Carmen Pampa. The objective of this study was to evaluate the effects of weaning male and female piglets at different times on their weight gain and the fertility of the sows. Four three year old sows that had a total of 40 piglets (25 males and 15 females) were used. The piglets were distributed across the following treatments: T1 - weaning 9 piglets (5 males and 4 females) at 56 days, T2 - weaning 13 piglets (7 males and 6 females) at 46 days, T3 - weaning 7 piglets (5 males and 2 females) at 36 days, and T4 - weaning 11 piglets (8 males and 3 females) at 26 days. This was followed by three weekly weight management phases in both sexes: Beginning (1-56 days), Growth (1-112 days), and Finished (1-150 days). A completely randomized design with different numbers of repetitions was used to analyze the results. The ANOVA test for weight gain did not show significant differences between treatments (p>0.05) in the beginning, growth, and finished stages for male piglets. The ANOVA test for weight gain in females during the initial phase showed significant differences (p<0.05). The Duncan test showed that T3 (12.50 kg) and T1 (10.80 kg) as well as T1 (10.80 kg) and T2 (9.41 kg) are equal. In addition, T2 (9.41 kg) and T4 (7.20 kg) are similar. In the growth phase there were no significant differences (p>0.05). In the fattening phase there were significant differences (p>0.05). The Duncan test showed that T3 (60.50 kg), T2 (56.42 kg), and T1 (54.75 kg) are equal and that T1 (54.75 kg) and T4 (43.33 kg) were similar at the end of the trial. The comparison of males and females was done via the student t test and indicated that there was no significant difference between males and females. The sow in T3 (44 days) recorded the best postpartum fertility. In general, treatment T3 (weaned at 36 days) showed the best results for both weight and fertility.

Bei den Fleischrinderrassen Angus, Fleckvieh und Limousin wurde untersucht, inwieweit Rasseunterschiede und biotechnische Verfahren den Erfolg der Künstlichen Besamung (KB) beeinflussen. Im Ergebnis sollen optimale Zeitspannen für die KB nach verschiedenen Hormonapplikationen abgeleitet werden. Bei 40 von insgesamt 88 besamten Tieren konnten mittels Ultraschalluntersuchung 34 Tage nach der KB Trächtigkeiten nachgewiesen werden. Der Besamungserfolg war bei Kühen etwas größer als bei Färsen, bei Fleckvieh etwas geringer als bei den anderen Rassen. Bei Tieren, deren Gelbkörper zum Zeitpunkt der PGF2α-Gabe einen Hohlraum aufweist, sollte die anschließende KB nur bei guter Brunstsymptomatik stattfinden. Hinsichtlich des KB-Termins erscheint eine Besamung bei gleichzeitiger GnRH-Gabe 68 bis 70 Stunden nach PGF2α-Injektion am sinnvollsten. Die Zeitspanne zwischen PGF2α-Injektion und Ovulation lag im Mittel bei 85 Stunden.

Durante o processo de criopreservação de sêmen, os espermatozóides sofrem alguns danos que resultam na diminuição da fertilidade deste. O presente estudo foi realizado a fim de avaliar o efeito da utilização, combinada de duas curvas de congelamento com dois diluentes comerciais sobre a criopreservação de sêmen eqüino. Foram analisados 20 ejaculados. As amostras foram avaliadas, pela motilidade progressiva e total do sêmen pós-descongelamento e pela integridade e funcionalidade da membrana dos espermatozóides. A combinação entre curva automatizada e Botu- Crio® apresentou as maiores médias, nas análises de motilidade total (55,53%) e progressiva (17,25%), após o descongelamento. O diluente Botu-Crio® ,isoladamente, apresentou também os melhores resultados quando foram realizadas as análises de integridade (CFDA/PI) e funcionalidade de membrana pelo teste hiposmótico.
During semen cryopreservation, sperm cells were submitted to some deleterious events leading to membrane damage which result in fertility decrease. This study was designed to compare the effects of two freezing techniques (conventional and automated), their freezing rates and the use of two commercial extenders as cryoprotectants (FR-5® and Botu-Crio®) on the total and progressive motility, integrity and functionality of spermatic membranes during the cryopreservation of equine semen. Twenty ejaculates were analyzed. The total and progressive motility of fresh and postthawing semen samples were evaluated by the patterns assays. The function of plasmatic membrane was measured by the hipoosmotic swelling test. The integrity of plasmatic membrane was evaluated using CFDA/PI fluorescent probes. There were significant differences between the two freezing techniques and/or between cryoprotectants for all assessed parameters. The combined used between Botu-Crio® and automated curves showed better results in total and progressive post-thawing motility. The extender Botu-Crio®, alone, showed better results in order to preserve the membrane integrity and function.

The European honey bee (Apis mellifera) has been present in Australia for approximately 150 years. For the majority of that time it was assumed this species could only be of benefit to Australia‘s natural ecosystems. More recently however, researchers and conservationists have questioned this assumption. Honey bees are an introduced species and may be affecting native fauna and flora. In particular, native bees have been highlighted as an animal that may be experiencing competition from honey bees as they are of similar sizes and both species require nectar and pollen for their progeny. Most research to date has focused on indirect measures of competition between honey bees and native bees (resource overlap, visitation rates and resource harvesting). The first chapter of this thesis reviews previous research explaining that many experiments lack significant replication and indirect measures of competition cannot evaluate the impact of honey bees on native bee fecundity or survival. Chapters two and four present descriptions of nesting biology of the two native bee species studied (Hylaeus alcyoneus and an undescribed Megachile sp.). Data collected focused on native bee fecundity and included nesting season, progeny mass, number of progeny per nest, sex ratio and parasitoids. This information provided a picture of the nesting biology of these two species and assisted in determining the design of an appropriate experiment. Chapters three and five present the results of two experiments investigating the impact of honey bees on these two species of native bees in the Northern Beekeepers Nature Reserve in Western Australia. Both experiments focused on the fecundity of these native bee species in response to honey bees and also had more replication than any other previous experiment in Australia of similar design. The first experiment (Chapter three), over two seasons, investigated the impact of commercial honey bees on Hylaeus alcyoneus, a native solitary bee. The experiment was monitored every 3-4 weeks (measurement interval). However, beekeepers did not agist hives on sites simultaneously so measurement intervals were initially treated separately using ANOVA. Results showed no impact of honey bees at any measurement interval and in some cases, poor power. Data from both seasons was combined in a Wilcoxon‘s sign test and showed that honey bees had a negative impact on the number of nests completed by H. alcyoneus. The second experiment (Chapter 5) investigated the impact of feral honey bees on an undescribed Megachile species. Hive honey bees were used to simulate feral levels of honey bees in a BACI (Before/After, Control/Impact) design experiment. There was no impact detected on any fecundity variables. The sensitivity of the experiment was calculated and in three fecundity variables (male and female progeny mass and the number of progeny per nest) the experiment was sensitive enough to detect 15-30% difference between control and impact sites. The final chapter (Chapter six) makes a number of research and management recommendations in light of the research findings.

L'objectif de l'etude est de preciser l'influence de la frequence de collecte du sperme et du type genetique du male sur les caracteristiques de la semence et la fertilite du lapin male. Trois experimentations sont realisees. Les deux premieres comparent, sur 52 et 14 semaines respectivement, 3 frequences de collecte: extensive (1 jour par semaine, le jeudi), intermediaire (mardi, vendredi), et intensive (lundi, mercredi, vendredi). La 3eme experience compare deux souches de males (a1077 et a2066) pendant 16 semaines. Chaque modalite experimentale comprend un effectif de l'ordre de 15 males. Des tests de fertilite par insemination sont realises au cours des 1ere et 3eme experiences. Le rythme extensif permet d'obtenir les moyennes par ejaculat les plus elevees de volume, de pourcentage de spermatozoides vivants, de motilite, de concentration et de nombre de spermatozoides vivants ou totaux. Par rapport aux frequences intermediaire et intensive, le rythme extensif ameliore le dernier caractere de +65% et +136% respectivement. Les caracteristiques de la semence ont une variabilite importante. La production hebdomadaire de spermatozoides est plus elevee avec le rythme intermediaire ou intensif, mais le gain enregistre ne depasse pas +30%. L'ecart de qualite de semence entre les rythmes extensif et intensif correspond a une difference de taux de fertilite (69,5% contre 57,2%) et de taille de portee (8,33 contre 6,71 nes totaux). Les moyennes des caracteristiques de l'ejaculat sont largement superieures dans la souche a1077, mais les taux de fertilite et tailles de portee des deux souches sont similaires. Toutefois, une analyse de correspondance montre l'existence d'une relation entre les caracteres de la semence et la fertilite ou la prolificite. Le rythme extensif de collecte permet de realiser la meilleure adequation entre les caracteristiques de l'ejaculat, la production hebdomadaire de spermatozoides, et la fertilite