The effects of allopurinol on the ultrastructure of ischaemic and reperfused large intestine of sheep

Objective To test the possible inhibitory effect of allopurinol on reperfusion injury, caused by oxygen-derived free radicals, of sheep large intestine.
Design An ultrastructural study on caecal tissues from control and treated groups.  

Animals

Fifty sheep in four ischaemic and reperfused (treatment) groups and one control group. Three of the treatment groups were subdivided for half to be injected with allopurinol and the other half with its solvent, potassium hydroxide (KOH).
Procedure Ischaemia of the caecum was induced in the four treatment groups for 60 minutes by clamping the apex. Allopurinol and its KOH solvent were injected intravenously in three treatment groups prior to ischaemia. Samples were collected before and 1 hour after induction of ischaemia and 1 min, 1 h and 8 h after reperfusion. Tissues were processed and examined with an electron microscope.
Results Untreated and solvent injected sheep showed minor ultrastructural changes following ischaemia. With reperfusion, there was severe mitochondrial, goblet cell and basement membrane damage. Tissues from allopurinoltreated sheep were preserved and appeared similar to tissues from the control group.  

Conclusion:

Pre-treatment with allopurinol prevented damage to tissues whereas untreated or allopurinol solventtreated showed severe damage following reperfusion. It is believed that allopurinol, an analogue of hypoxanthine and xanthine, prevents reperfusion injury by competitively binding with xanthine oxidase. This reduces or inhibits the xanthine oxidase mediated conversion of hypoxanthine to xanthine thereby preventing the formation of oxygen-derived free radicals.     

Viability of OPS Vitrified Sheep Embryos After Direct Transfer

The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions.     

Two Methods of Vitrification Followed by In Vitro Culture of the Ovine Ovary: Evaluation of the Follicular Development and Ovarian Extracellular Matrix

The aim of this study was to evaluate the influence of two vitrification techniques on the extra cellular matrix (ECM) and ovarian follicular development. The ovarian cortex was fragmented (9 mm³) and divided into six groups, viz. fresh control, cultured control, vitrified by the Ovarian Tissue Cryosystem (OTC) method, conventional solid surface vitrification (SSV) method, OTC/cultured and SSV/cultured. Follicles from all the fragments were analysed for morphology, development and viability. The ECM was evaluated based on the condition of collagen and reticular fibres and the immunolocalization of type I collagen and fibronectin. After 7 days of culture, the tissue vitrified by OTC revealed a higher percentage (p < 0.05) of morphologically normal (30.66%) and viable (60.00%) follicles when compared with those vitrified using the SSV technique (21.33% and 23.00%). In all the fragments cultured, regardless of the vitrification method, a significantly higher percentage of developing follicles was observed when compared with the non‐cultured tissue. Analysis of the type I collagen showed increased immunostaining after the in vitro culture in the vitrified fragments. In conclusion, the OTC is better for preserving the follicular viability and morphology and maintaining the integrity of the extracellular matrix components of the ovine ovary.     

Fibrin–alginate hydrogel supports steroidogenesis,in vitro maturation of oocytes and parthenotes production from caprine preantral follicles cultured in group

This study aimed to establish a culture system that improves the in vitro development of caprine preantral follicles. In a first experiment, follicles were encapsulated as a single unit per bead and cultured singly or in groups or with five follicles in the same alginate (ALG) bead for 18 days. In a subsequent experiment, the “five follicles per bead” design was chosen to culture in ALG, fibrin–alginate (FA) or hyaluronate (HA) for 18 days. In a third experiment, we chose the five follicles per bead in FA to culture for 30 days. The culture set‐up of five follicles per ALG bead increased antrum formation and follicle diameter compared to the other culture designs (p < .05). Moreover, under this condition, 44.44% of the oocytes from in vitro cultured preantral follicles reached meiotic resumption. A significant increase of follicle diameter occurred in attachment system and FA (p < .05), but the ALG condition reached the highest among all groups on day 18 (p < .05). Follicles encapsulated in matrix produced more estradiol and progesterone than attachment system (p < .05). The expression of MMP‐9 mRNA was higher in FA than in other groups (p < .05) and similar to antral follicles from in vivo control (p > .05). Only FA group resulted in oocytes matured. After 30 days, oocytes from preantral follicles in vitro grown in FA developed to eight‐cell parthenotes. In conclusion, a culture system using FA supported the development of caprine preantral follicles cultured in group and included in the same bead of hydrogel, improving the oocyte maturation and producing parthenotes.     

Efficacy of maduramicin ammonium against coccidiosis in turkeys under laboratory and floor-pen conditions

Experimental infections with field isolates of Eimeria meleagrimitis, E. adenoeides, E. gallopavonis, and E. dispersa in turkey poults were used to test the efficacy of maduramicin ammonium at 2.5-10 ppm in laboratory experiments. Infection with single or mixed species of coccidia reduced the weight gain of unmedicated infected controls and caused 18.1-65% mortality in two experiments. Maduramicin ammonium given at 5-7 ppm prevented mortality, significantly reduced droppings scores and oocyst passage, and improved weight gain to near that of the unmedicated uninfected controls. Maduramicin ammonium was tested at 4-7 ppm in a floor-pen trial lasting 10 weeks. Mortality from coccidiosis averaged 11.9% in unmedicated controls, compared with 0.6% with 4 ppm of maduramicin or no mortality with 5-7 ppm. Average weight gain and feed conversion at 10 weeks were significantly improved over unmedicated infected controls when maduramicin ammonium was given at 5-7 ppm. These results suggest that maduramicin ammonium is highly efficacious against field isolates of Eimeria in turkeys, especially within the range of 5-7 ppm in the feed.     

Chemical Activation with a Combination of Ionomycin and Dehydroleucodine for Production of Parthenogenetic,ICSI and Cloned Bovine Embryos

The aim of this study was to evaluate the potential of dehydroleucodine (DhL), a new drug isolated from a medicinal herb used in Argentina, for activation of bovine oocyte. Several DhL concentrations and exposure times after ionomycin (Io) treatment were tested. The optimal DhL treatment, found for parthenogenetic development, was employed to produce bovine embryos by intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). The best parthenogenic embryo developments were observed with 5 μm Io for 4 min followed by 5 μm DhL concentration and after 3‐h exposure time (52.3% cleavage; 17.4% morulae; 7.3% blastocyst; n = 109). This treatment generated no significant differences with standard Io plus 6‐dimethylaminopurine (DMAP) treatment in preimplantation embryo development. In our conditions, the embryo development reached after ICSI and SCNT assisted by the DhL treatment did not differ in terms of cleavage and blastocyst development from activation with standard Io plus DMAP treatment (p > 0.05). In conclusion, DhL utilization to activate oocytes and induce development of parthenogenotes, ICSI‐embryos or SCNT‐embryos is reported here for first time.     

The effects of day and night temperature on Chrysanthemum morifolium: investigating the safe limits for temperature integration

Summary

The impact of day and night temperatures on pot chrysanthemum (cultivars ‘Covington’ and ‘Irvine’) was assessed by exposing cuttings, stuck in weeks 39, 44, and 49, to different temperature regimes in short-days. Glasshouse heating set-points of 12°, 15°, 18°, and 21°C, were used during the day, with venting at 2°C above these set-points. Night temperatures were then automatically manipulated to ensure that all of the treatments achieved similar mean diurnal temperatures. Plants were grown according to commercial practice and the experiment was repeated over 2 years. Increasing the day temperature from approx. 19°C to 21°C, and compensating by reducing the night temperature, did not have a significant impact on flowering time, although plant height was increased. This suggests that a temperature integration strategy which involves higher vent temperatures, and exploiting solar gain to give higher than normal day temperatures, should have minimal impact on crop scheduling. However, lowering the day-time temperature to approx. 16°C, and compensating with a warmer night, delayed flowering by up to 2 weeks. Therefore, a strategy whereby, in Winter, more heat is added at night under a thermally-efficient blackout screen may result in flowering delays. Transfers between the temperature regimes showed that the flowering delays were proportional to the amount of time spent in a low day-time temperature regime. Plants flowered at the same time, irrespective of whether they were transferred on a 1-, 2-, or 4-week cycle.     

Horizontal movements,utilization distributions,and mixing rates of yellowfin tuna (Thunnus albacares) tagged and released with archival tags in six discrete areas of the eastern and central Pacific Ocean

A total of 1522 yellowfin tuna, Thunnus albacares, were captured, tagged, and released with surgically implanted archival tags (ATs), in six discrete areas of the eastern and central Pacific Ocean, during 2002 through 2019. Of 483 ATs returned (31.7%), 227 ATs from yellowfin (48–147 cm in fork length) at liberty from 32 to 1846 d ( = 300.1 d) provided suitable data sets which were processed using an unscented Kalman filter model with sea-surface temperature measurements integrated (UKFsst) in order to obtain most probable tracks and movement parameters. Although some differences were observed in the movement patterns for fish from within and among the six release areas, 99% of the 227 fish remained within 1000 M of their release locations, indicating limited dispersion and fidelity to release locations. The median movement parameter D, which defines dispersion from the UKFsst model, for the fish released in the offshore equatorial areas showed much greater dispersion rates compared to those for the fish released along the coast or around islands. The rates of mixing of yellowfin among the release areas were found to be dependent on the distances between release areas, with, in general, the greatest mixing occurring among areas in closest proximity, whereas for the two areas offshore Mexico and the two offshore equatorial areas, the rates of mixing were nonexistent or negligible.