Lectin histochemical pattern on the normal and cystic ovaries of sows

The lectin histochemical pattern (LHP) was characterized and compared in normal and cystic ovaries of sows. Six biotinylated lectins (PNA, SBA, WGA, RCA‐1, DBA and UEA‐1) were used on tissue sections. In the normal ovaries, the reaction to UEA‐1 and SBA was mild to moderate in mesothelial and endothelial cells. RCA‐1 staining was mild to moderate in theca interna of growing follicles, corpora luteum and mesothelium. In addition, this lectin presented strong reaction in endothelial cells, granulosa cells of atretic follicles, zona pellucida of growing follicles and plasma. DBA showed strong intensity in mesothelial and endothelial cells. There was mild to moderate reactivity to WGA in granulosa cells, corpus luteum and theca interna of follicles in development, and moderate in zona pellucida, in granulosa cells of atretic follicles and mesothelium. PNA staining was mild to moderate in oocytes and in the adventitia and media of medullary arteries. Changes in the LHP of the cystic ovaries were noted; however, there were no differences in these findings between the follicular and luteinized cysts. UEA‐1 reactivity in the cystic ovaries was moderately reduced in the mesothelial and endothelial cells, whereas there was mild reduction in the DBA staining in the granulosa cells. Reaction to RCA‐1 and WGA in the cysts also was decreased in theca interna, zona pellucida and granulosa cells of atretic follicles. Furthermore, endothelium and theca interna in the cystic ovaries presented mild reduction of marcation to SBA, whereas there was decreased reactivity to PNA in the oocytes and adventitia and media layers of the medullary arteries. The results of the current study show that cysts modify the LHP in swine ovaries. These changes of glycoconjugates in many ovarian structures could modify diverse process and may be one of the reasons for decreased fertility in sows.     

Changes in the concentrations of auxins and polyamines during rooting of in-vitro-propagated walnut shoots

Rooting was induced in in-vitro-propagated walnut (Juglans regia L.) shoots by subculturing the shoots on rooting medium containing agar and 3 mg l(-1) indole-3-butyric acid (IBA) for 7 days in darkness. Changes in the concentrations of endogenous free indole-3-acetic acid (IAA), indole-3-acetylaspartic acid (IAAsp) and free polyamines were determined during culture on root-inducing medium. In extracts of whole shoots, the concentration of free IAA showed a transient peak at 60 h (around 48 h in extracts from basal shoot portions) and then remained at a relatively low concentration for the remainder of the 7-day culture period. The concentration of IAAsp in extracts of whole shoots peaked at about the same time as the concentration of free IAA, whereas the IAAsp concentration in extracts from basal shoot portions peaked earlier, at around 12 h. The concentrations of free polyamines in extracts of whole shoots increased soon after the shoots were transferred to root-inducing medium. The concentrations of IAA and IAAsp remained stable when the rooted shoots were transferred to a vermiculite/gelrite mixture (without auxin) and grown in light.     

Anomalous Pregnancies during Late Embryonic/Early Foetal Period in High Producing Dairy Cows

This study analyses anomalous cases of gestation ending in pregnancy loss during the early foetal period and their effect on progesterone and plasma pregnancy-associated glycoprotein-1 (PAG-1) concentrations. Data derived from a large-scale ultrasound pregnancy diagnosis programme in high producing dairy cows. Over a 3-year period (2004–2007), a very low incidence (0.5%: 15 of 3094) of anomalous pregnancies was recorded. The results revealed that the following anomalies were detected on days 35–41 of gestation in cows carrying singletons with one single corpus luteum: embryo death in eight cows (0.3%); and embryo in the uterine horn contralateral to the corpus luteum in seven cows (0.2%). All these animals suffered pregnancy loss during the early foetal period. In cows carrying dead embryos, no signs of conceptus degeneration were observed on pregnancy diagnosis. Amnion size (approximately 25 mm diameter) and uterine horn fluid contents were estimated to be similar to those of the normal pregnant cows in this period. In the contralateral gestations, live embryos were observed in all ultrasound checks before pregnancy loss. Uterine fluid contents increased in the two cows in which gestation continued for more than a week. In the cases of embryo death but not in those of contralateral gestation, a drop in PAG-1 levels was noted prior to pregnancy loss. Two cows carrying dead embryos increased with time allantoic fluid contents. The PAG-1 values increased with time in one cow bearing a dead embryo (from 2.31 to 6.79 ng/ml) and in two of the contralateral gestations (from 1.66 to 2.33 ng/ml and from 0.39 to 6.79 ng/ml, respectively). Results of this study indicate that the foetal membranes continue to undergo some activity following embryo death, and that contralateral pregnancy may determine failure of the gestation process.     

Serum Profiles of Pregnancy-Associated Glycoprotein, Oestrone Sulphate and Progesterone During Gestation and Some Factors Influencing the Profiles in Ethiopian Borana and Crossbred Cattle

This study presents serum concentrations of pregnancy‐associated glycoprotein (PAG), oestrone sulphate (E1‐S) and progesterone (P4), and the effects of some dam and foetus‐related factors on these profiles during gestation in Borana and crossbred cattle. The PAG concentrations at 4th week post‐conception ranged from 1.5–5.5 and 2.1–4.7 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The mean PAG concentrations increased progressively from 4th to 33rd week of gestation (from 3.3–173 ng/ml for Borana and 4.2–240 ng/ml for crossbred cattle) and reached peak around calving. Breed, parity status, dam body weight, foetal sex and foetal birth weight significantly influenced the PAG concentrations. After delivery, the PAG concentrations declined steadily to 5.7 ng/ml in Borana (n = 7) and 3.9 ng/ml in crossbred (n = 6) cattle 10 weeks post‐partum. The serum E1‐S concentrations at 17th week of pregnancy ranged from 0.3–2.6 and 0.9–5.7 ng/ml in Borana (n = 8) and crossbred (n = 9) cattle, respectively. The mean E1‐S concentrations increased progressively from 17th to 33rd week of gestation (from 1.1–4.6 ng/ml for Borana and 2.7–10.8 ng/ml for crossbred). Breed, parity status, dam body weight and foetal sex significantly influenced E1‐S concentrations. The P4 concentrations at 4th week of pregnancy ranged from 3.2–5.1 and 1.7–8.9 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The P4 level remained elevated throughout pregnancy. This study indicated that the serum PAG and P4 concentrations at 4th and E1‐S approximately 17th week of pregnancy were above the cut‐off value for pregnancy test and the hormonal profiles observed were comparable to the previous reports. Furthermore, the PAG and E1‐S profiles were considerably influenced by factors such as breed, weight and parity status of the dam, and foetal sex and foetal birth weight (only PAG).     

Pre-breeding Ovaro–Uterine Ultrasonography and its Relationship with First Service Pregnancy Rate in Seasonal-Calving Dairy Herds

The objectives of the study were to characterize an ultrasound reproductive tract scoring (URTS) system to assess suitability for breeding in dairy cows, to describe the prevalence of these scores in commercial dairy herds and to examine their relationship with subsequent fertility. Ultrasound examinations (7797) were performed on 5751 Holstein–Friesian cows prior to breeding in 62 seasonally calving herds over 2 years. Data recorded from images of both ovaries and the uterus were combined into a six point scoring system and the prevalence of cows with cystic ovarian follicles and uterine abscesses and adhesions was recorded separately. The prevalence of ovulatory cows which had completed uterine involution (score 1), or had mild (2) or moderate endometritis (3) was 62.5%, 21.7% and 2.2%, respectively. The prevalence of anovulatory cows with moderate endometritis (4), ovulatory cows with pyometra (5) and anovulatory cows which had completed uterine involution (6) was 3.3%, 2.2% and 8.1%, respectively. The interval between calving and examination differed between cows with each of the scores 1, 2, 5 and 6 (61, 46, 53 and 50 days, respectively, p < 0.05) but not between cows with scores 3 and 4 (37 and 35 days, respectively). Ovulatory cows which had completed uterine involution (score 1) had a higher likelihood of pregnancy to first service than ovulatory or anovulatory cows which had not completed uterine involution (p ≤ 0.10). The prevalence of cows with cystic ovarian follicles or abscesses or adhesions of the reproductive tract was 3.9% and 1.2%, respectively. In conclusion, 29% and 11% of cows in seasonally calving and breeding dairy herds had not completed uterine involution or were anovulatory prior to the mating start date, respectively. Both conditions, detected using a URTS system, significantly reduced first service pregnancy rate in these pasture-based dairy herds.     

Pregnancy‐Associated Glycoprotein Secretion in North Moroccan Goats

The plasma profile of pregnancy‐associated glycoprotein (PAG) and its relationship with fetal number were studied in 14 North Moroccan goats using a semi‐heterologous radioimmunoassay (RIA). Peripheral blood was collected every day during the first month of pregnancy, afterward the blood samples were collected trice a week. The PAG were first detected at day 20 of pregnancy, their levels increase to week 8 of gestation then decrease slightly until parturition. Statistical differences between goats carrying one or two foetuses are observed from week 5 of pregnancy until parturition. Factorial Discriminant Analysis provides mathematical models for the discrimination between the litter size using the PAG level and the week of pregnancy. Using only one blood sample per week, high sensitivity, specificity and overall accuracy (74%, 88% and 81%) were obtained using these models. It is concluded that the PAG RIA is an effective tool for early diagnosis of pregnancy and for discrimination between the litter sizes in North Moroccan goats.     

Energy Metabolism and Leptin: Effects on Neuroendocrine Regulation of Reproduction in the Gilt and Sow

It is well established that reproductive function is metabolically gated. However, the mechanisms whereby energy stores and metabolic cues influence appetite, energy homeostasis and fertility are yet to be completely understood. Adipose tissue is no longer considered as only a depot to store excess energy. Recent findings have identified numerous genes, several neurotrophic factors, interleukins, insulin-like growth factor binding protein-5, ciliary neurotrophic factor and neuropeptide Y (NPY) as being expressed by adipose tissue during pubertal development. These studies demonstrated for the first time the expression of several major adipokines or cytokines in pig adipose tissue which may influence local and central metabolism and growth. Leptin appears to be the primary metabolic signal and is part of the adipose tissue-hypothalamic regulatory loop in the control of appetite, energy homeostasis and luteinizing hormone (LH) secretion. Leptin's actions on appetite regulation are mediated by inhibition of hypothalamic NPY and stimulation of proopiomelanocortin. Its effects on gonadotropin-releasing hormone (GnRH)/LH secretion are mediated by NPY and kisspeptin. Thus, leptin appears to be an important link between metabolic status, the neuroendocrine axis and subsequent fertility in the gilt and sow.     

Metabolic responses to water deficit in two Eucalyptus globulus clones with contrasting drought sensitivity

We compared the metabolic responses of leaves and roots of two Eucalyptus globulus Labill. clones differing in drought sensitivity to a slowly imposed water deficit. Responses measured included changes in concentrations of soluble and insoluble sugars, proline, total protein and several antioxidant enzymes. In addition to the general decrease in growth caused by water deficit, we observed a decrease in osmotic potential when drought stress became severe. In both clones, the decrease was greater in roots than in leaves, consistent with the observed increases in concentrations of soluble sugars and proline in these organs. In roots of both clones, glutathione reductase activity increased significantly in response to water deficit, suggesting that this enzyme plays a protective role in roots during drought stress by catalyzing the catabolism of reactive oxygen species. Clone CN5 has stress avoidance mechanisms that account for its lower sensitivity to drought compared with Clone ST51.     

Effect of Sperm Cryopreservation on the European Eel Sperm Viability and Spermatozoa Morphology

The main objective of the present work was to study the effect of cryopreservation of European eel sperm both on the sperm viability and the spermatozoa head morphology. Spermatozoa morphology was evaluated with computer-assisted morphology analysis after collection in fresh samples, after adding the freezing medium containing dimethyl sulfoxide as cryoprotectant and, finally, after the cryopreservation process and thawing. Cell viability was assessed, in both fresh and thawed samples, by Hoechst 33258 staining. Computer-assisted sperm analysis (CASA) was used to determine the percentage of motile cells and to measure motility parameters in sperm samples. A significant decrease of head perimeter (12.56%) and area (17.90%) was detected from spermatozoa in fresh to thawed samples, indicating that cells do not recover the original size after the cryopreservation process. CASA was used to measure the percentage of motile cells (51.9%) and spermatozoa motility parameters such as curvilinear, straight line and angular path velocities, as well as beating cross frequency. This technique was employed in the fresh sperm samples but proteins present at the freezing medium (L-alpha-phosphatidylcholine) made impossible to use this last technique in thawed samples. When sperm viability was assessed by Hoechst staining, a significant decrease of approximately 15% (73.10 vs 58.26%) of alive spermatozoa was registered from fresh to thawed samples. The percentage of motile cells measured by CASA in fresh samples (51.9%) was lower than the percentage of alive cells determined by Hoechst stainning, suggesting the existence of different batches of spermatozoa in different stages of development, even during the eight to tenth weeks of treatment, when the highest sperm quality was found.