Comparative IFN-τ Secretion after Hatching by Bovine Blastocysts Derived Ex Vivo and Completely Produced In Vitro

The interferon-tau (IFN-tau) secretion levels after hatching by bovine blastocysts derived from in vitro maturated oocytes (Group A) and from in vivo (Group B) were investigated considering embryo quality. Only very homogeneous blastocysts of excellent or good quality were considered from day 7 of culture (Group A) and day 7 after artificial insemination with frozen-thawed from the same bull used for in vitro fertilization (Group B). All embryos were individually cultured into a 50 microl droplet of synthetic oviduct fluid medium with 10% fetal calf serum. After 24-h culture both Group A (n=44) and B (n=40) secreted <54 pm IFN-tau. After 48-, 72-, 96- and 120-h culture, Group A daily secreted 143 +/- 24 pm IFN-tau (n=19) vs 85 +/- 12 pm IFN-tau (n=21) for Group B (p < 0.01), 491 +/- 128 pm IFN-tau (n=29) vs 216 +/- 37 pm IFN-tau (n=23) (NS), 499 +/- 135 pm IFN-tau (n=26) vs 353 +/- 93 pm IFN-tau (n=21) (NS), 559 +/- 136 pm IFN-tau (n=22) vs 333 +/- 75 pm IFN-tau (n=20) (NS), respectively. Taken all together during 5 days, Group A produced per embryo 1690 +/- 290 pm IFN-tau (n=22) vs 982 +/- 182 pm IFN-tau (n=20) for Group B (p < 0.05). For all culture time there were sizable percentages of embryos that did not produce concentrations of IFN-tau above a certain cut-off level, and as such were not used to compute the means. In respect of the embryo quality whatever the groups after days 7-12 of culture, IFN-tau secretions were 1815 +/- 453 pm (n=10) for the embryos of excellent quality vs 1356 +/- 200 pm (n=28) for those of good quality (NS) and 360 +/- 188 pm (n=4) (p < 0.05) for embryos of fair quality. A positive relationship between IFN-tau production and in vitro development of quality I embryos was observed, whatever the embryos origins and, the embryos completely produced in vitro secreted more IFN-tau than the embryos produced in vivo.     

Progesterone and Caspase-3 Activation in Equine Cyclic Corpora Lutea

Soon after ovulation, the newly formed corpus luteum (CL) starts secreting progesterone (P(4)), necessary for implantation. The CL, an ovarian transient endocrine organ, undergoes growth and regression throughout its life span. The objective of this study was to evaluate if caspase-3 mediates cell death in the equine cyclic luteal structures and relate it to luteal endocrine function. Blood and luteal tissue were collected during the breeding season after slaughter from 38 randomly assigned cycling mares. Luteal tissues were classified as corpora haemorrhagica (CH; n = 7); mid luteal phase corpora lutea (Mid-CL; n = 17); late or regressing corpora lutea (Late-CL; n = 9) and corpora albicans (CA; n = 5). Plasma P(4) concentration, determined by radioimmunoassay, showed a significant increase from CH to Mid-CL (p < 0.001), followed by a decrease to Late-CL (p < 0.001) and CA (p < 0.001). Caspase-3 processing and poly (ADP) ribose polymerase (PARP) degradation were assessed by western blotting. Active caspase-3 was twofold increased in Mid-CL, Late-CL and CA as compared with CH (p < 0.05). Immunocytochemistry also showed a significant increase in caspase-3 expression in large luteal cells in all structures when compared with CH (p < 0.05). Consistently, the endogenous caspase-3 substrate, PARP, was markedly degraded from CH to CA (p < 0.05). In fact, the ratio of full-length to degraded PARP showed a significant decrease from CH to Mid-CL, Late-CL and CA (p < 0.05). Finally, the decrease in P(4) from Mid- to Late-CL coincided with no further increases in apoptosis. In conclusion, these results suggest that the effector caspase-3 of apoptosis, might play an important role during luteal tissue involution in the mare, even though its relationship with P(4) remains to be elucidated.     

Management Strategies Aiming to Improve Horse Welfare Reduce Embryonic Death Rates in Mares

The objective of this retrospective study was to evaluate the effect of management strategies aiming to improve animal well‐being on pregnancy and embryonic death (ED) rates. Breeding records of a cohort of 1206 Thoroughbred mares brought to a stallion station facility, to be bred with the stallions housed there, were evaluated during ten breeding seasons. Mares were blocked according to management strategies in two groups: Stress and Relax. Strategies used to improve animal well‐being (Relax group) were as follows: stopping the teasing routine, reducing or eliminating stall confinement, reducing the number of mares per group and maintaining herd stability during the breeding season. In barren mares, the pregnancy rate was higher in the Relax group (91.8%) when compared to the observed in Stress group (84.7%). However, no difference in pregnancy rates were observed (Stress = 85.2% vs. Relax = 86.2) in foaling mares. ED rate was higher in barren and foaling mares of the Stress group mares (25.5% and 26.8%, respectively) compared with the Relax group (16.1% and 14.7%, respectively). No significant differences were observed on foal heat pregnancy rate between groups; yet, the embryo loss on foal heat was significant reduced in Relax mares (Relax = 8.7% vs Stress = 24.5%). In conclusion, management strategies aimed to reduce social stress can reduce early pregnancy losses and the average cycles per pregnancy, improving reproductive performance in mares.     

Influence of dietary endophyte (Neotyphodium coenophialum)-infected tall fescue (Festuca arundinacea) seed on fecal shedding of antibiotic resistance-selected Escherichia coli O157:H7 in ewes

The objectives were to determine the effects of short-term feeding of a toxic endophyte (Neotyphodium coenophialum)-infected tall fescue seed (Festuca arundinacea, cultivar 'Kentucky 31') on fecal shedding and intestinal concentrations of Escherichia coli O157:H7 and the concentrations of prolactin, cortisol, and NEFA in experimentally inoculated ewes. Twelve ewes (mean BW = 46 +/- 2 kg) were fed a diet containing either high endophyte-infected (HI-E) or low endophyte-infected (LO-E) tall fescue seed for 7 d. Each diet consisted of 50% (as-fed basis) tall fescue seed. Ewes were experimentally inoculated with antibiotic resistance-selected E. coli O157:H7 on d 1 of the feeding treatment, and fecal shedding of inoculated pathogens was monitored daily on d 2 to 6. On d 7, ewes were weighed and euthanized, and tissues and contents were sampled from the ileum, cecum, and rectum for quantitative enumeration of E. coli O157:H7. Urine was collected at euthanization to determine total ergot alkaloid concentrations. Ewes fed HI-E had lower (P < 0.001) DMI than did ewes fed LO-E (0.8 and 1.6 +/- 0.1 kg/d of DMI for HI-E and LO-E ewes, respectively); consequently, there was a tendency (P = 0.06) for HI-E ewes to lose 0.3 +/- 0.4 kg of BW/d and LO-E ewes to gain 0.2 +/- 0.4 kg of BW/d during the 7 d. Urinary ergot alkaloids were increased (P < 0.001) in ewes fed HI-E (47.8 +/- 9.4 ng/mg of creatinine) compared with those fed LO-E (6.2 +/- 9.4 ng/mg of creatinine). Prolactin tended (P = 0.06) to be decreased in ewes fed HI-E (7.2 +/- 7.0 ng/mL) compared with those fed LO-E (27.7 +/- 7.0 ng/mL). Fecal shedding of E. coli O157:H7 tended (P = 0.06) to be increased in HI-E ewes [5.4 cfu (log10)/g of feces] compared with LO-E ewes [4.5 cfu (log10)/g of feces]. The population of E. coli O157:H7 in luminal contents from the ileum, cecum, and rectum did not differ (P > 0.36) between treatments. Treatment did not influence (P = 0.30) the occurrence of E. coli O157:H7 in cecal or rectal tissues; however, ileal tissues from HI-E ewes tended (P = 0.12) to have an increased incidence of E. coli O157:H7. Concentrations of NEFA tended (P = 0.12) to be greater in HI-E ewes than in LO-E ewes, whereas cortisol was similar (P = 0.49) for HI-E and LO-E ewes. We conclude that short-term feeding of HI-E tall fescue seed may alter the concentrations of prolactin and NEFA, and may increase fecal shedding of E. coli O157:H7 in experimentally inoculated ewes.     

Visualisation of <Emphasis Type="Italic">hrp</Emphasis> gene expression in <Emphasis Type="Italic">Xanthomonas euvesicatoria</Emphasis> in the tomato phyllosphere

The plasmid pUFZ75 conferred constitutive GFP expression on the bacterial pathogen Xanthomonas euvesicatoria (syn. X. campestris pv. vesicatoria). Colonisation of the tomato phyllosphere and invasion of tomato leaves by X. euvesicatoria was examined using both fluorescence and confocal laser scanning microscopy. Xanthomonas euvesicatoria established a limited population on the tomato leaf surface, primarily occupying the depressions between epidermal cells and around the stomata, prior to invasion of the leaf via the stomata and subsequent growth within the substomatal chamber and the leaf apoplast. Additionally, hrp-gfp fusions were used to report on the temporal and spatial expression of hrp genes during epiphytic colonisation and invasion. Xanthomonas euvesicatoria cells carrying hrpG- and hrpX-gfp reporter constructs were not fluorescent in vitro on non-hrp-inducing LB agar but did exhibit a low level of fluorescence on the leaf surface within 24 h of inoculation, particularly in the vicinity of stomata. Cells carrying hrpG- and hrpX-gfp fusions exhibited high levels of fluorescence 72 h after inoculation in the substomatal chamber and the leaf apoplast. Cells carrying the hrpF-gfp fusion were slightly fluorescent on LB agar and showed no further increase in fluorescence on the leaf surface by 24 h after inoculation, but did show a significant increase in fluorescence 72 h after inoculation in the substomatal chamber and apoplast. The apparent low-level induction of the regulators hrpG and hrpX on the tomato leaf surface may suggest that some of the genes of the X. euvesicatoria HrpG/HrpX regulon are up- or down-regulated prior to invasion of the stomata while still on the leaf surface.     

High-selectivity, high-flux silica membranes for gas separation

Process improvements in silica membrane fabrication, especially the use of clean-room techniques, resulted in silica membranes without detectable mesoscopic defects, resulting in significantly improved transport properties. Supported membranes calcined at 400 degreesC were 30 nanometers in thickness, showed a H2 permeance at 200 degreesC of 2 x 10(-6) moles per square meter per second per Pascal (mol m-2 s-1 Pa-1), and had a CH4 permeance more than 500 times smaller. Molecules larger than CH4 were completely blocked. Silica membranes calcined at 600 degreesC showed no detectable CH4 flux, with a H2 permeance of 5 x 10(-7) (mol m-2 s-1 Pa-1) at 200 degreesC. These results signify an important step toward the industrial application of these membranes such as purification of H2 and natural gas as well as the selective removal of CO2.     

Allelopathic potential of native plants against water hyacinth

Classical biocontrol of water hyacinth using insects in India is constrained by seasonal occurrence of water flow and interrupted host range. Use of fungal pathogens is also difficult due to lack of shelf-life and virulence under severe climatic fluctuations and a lack of knowledge of spray techniques. Hence, the allelopathic potential of native plants is reviewed for use as an alternative bio-control tactic. Dried powder of the leaves of Omavalli Coleus amboinicus L. at 40 g l⁻¹ as a water suspension killed water hyacinth within 24 h reducing the fresh weight by 80.72% and the dry weight by 75.63% within one week. The lowest dose required to kill the whole plant was 10 g l⁻¹. Coleus powder was injurious to cut leaves of water hyacinth even at 0.1 g l⁻¹ dose as it was absorbed directly by the cut leaf, indicating that the dose required to kill the whole plant could still be reduced, if either the natural product or the active principle is formulated for absorption through foliage.