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.     

Therapeutic Ultrasound as a Potential Male Dog Contraceptive: Determination of the Most Effective Application Protocol

Ultrasound is one of the most promising forms of non‐invasive contraception and has been studied in several animal models. The objective of the current investigation was to determine the most practical and effective application protocol for dog sterilization. A total of 100 dogs were divided into five equal groups. Group A received 5‐min applications three times performed at 48‐hr intervals and covering the entire testicular area at frequency of 1 MHz; Group B received 5‐min applications three times performed at 48‐hr intervals over the dorso‐cranial area of the testis at frequency of 3 MHz; Group C received three sequential 5‐min applications (at 5‐min intervals between applications) covering the entire testicular area at frequency of 1 MHz; Group D received 15‐min applications two times performed at 48‐hr intervals and covering the entire testicular area at frequency of 1 MHz. The experimental groups' ultrasound had an intensity of 1.5W/cm². The Control Group had the same procedure as Group A, but with the transducer switched‐off. Dogs were surgically castrated 40 days following the treatment for histological examination. Azoospermia, testicular volume reduction and apparently irreversible testicular damage were achieved by Group A. No effects were noticed in the other groups. Testosterone levels remained within physiological range with all application protocols. A regimen of three applications of ultrasound at 1 MHz, and 1.5 W/cm², lasting 5 min with an interval of 48 h was effective as permanent sterilization in the dog without hormonal impact.     

The dynamics of root production and loss in Betula papyrifera seedlings in response to elevated CO2 and an aluminium pulse

Seedlings of Betula papyrifera were grown in sand/nutrient solution cultures in rhizotrone growth containers under elevated (700 ppm) or ambient (375 ppm) atmospheric CO₂ concentrations for approximately 10 weeks. Thirty seven days after the begin of the experiment the plants were exposed to a 10 day pulse of 400 or 1200 μM Al. Elevated atmospheric CO₂ increased both root production and loss. Exposure to Al reduced root production and slightly reduced root loss. The reduced root production due to Al was amplified after the pulse had receded, resulting in a significantly lower net and gross root production at the end of the experiment. There were no clear CO₂ × Al interactions.     

Design and fabrication of topologically complex, three-dimensional microstructures

Two concepts for use in the fabrication of three-dimensional (3D) microstructures with complex topologies are described. Both routes begin with a two-dimensional (2D) pattern and transform it into a 3D microstructure. The concepts are illustrated by use of soft lithographic techniques to transfer 2D patterns to cylindrical (pseudo-3D) substrates. Subsequent steps-application of uniaxial strain, connection of patterns on intersecting surfaces-transform these patterns into free-standing, 3D, noncylindrically symmetrical microstructures. Microelectrodeposition provides an additive method that strengthens thin metal designs produced by patterning, welds nonconnected structures, and enables the high-strain deformations required in one method to be carried out successfully.     

Elevated atmospheric CO(2) concentration changes ectomycorrhizal morphotype assemblages in Betula papyrifera

Ectomycorrhizae are extremely diverse, with different species of fungi having very different physiologies and morphologies that, in turn, confer a range of different benefits to the host plant. To test the hypothesis that elevated CO(2) leads to changes in the assemblage of ectomycorrhizae associated with trees, we examined the number and frequency of ectomycorrhizal morphotypes colonizing roots of Betula papyrifera Marsh. saplings grown at an ambient or elevated (700 ppm) atmospheric CO(2) concentration for 24 weeks. Elevated CO(2) resulted in significant changes in the composition of the ectomycorrhizal assemblage toward morphotypes with a higher incidence of emanating hyphae and rhizomorphs. We conclude that B. papyrifera saplings will be able to support a more costly mycorrhization in future elevated-CO(2) atmospheres.     

Natural infection of broiler breeder chickens with endemic apathogenic Newcastle disease virus and their subsequent response to vaccination with a live V4 Newcastle disease virus vaccine

Flocks of broiler breeder chickens housed on a commercial farm were monitored from 13 w of age for natural infection with endemic lentogenic Newcastle disease virus (NDV). Seroconversion was first detected at 17 w. By 24 w, all 8 flocks had achieved peak log2 mean haemagglutination inhibiting antibody titres of up to 4.8. Antibody titres then declined and rose again over several months, suggesting cyclic reinfection with NDV. A lentogenic NDV indistinguishable from V4 was isolated from the cloaca of one bird at 18 weeks of age. At 54 weeks of age, 6 of 8 flocks were vaccinated en masse with live V4 NDV vaccine, 3 flocks by drinking water and 3 flocks by aerosol. All flocks were serologically monitored for a further 8 w. Drinking water vaccination induced an anamnestic response in 3 flocks, showing that flocks with pre-existing active immunity to NDV may be successfully vaccinated with V4. However, in all aerosol vaccinated flocks, the procedures failed to induce a response different to that observed in unvaccinated flocks. The serological response to vaccination was greater in sires than in dams.     

Effectiveness of α-tocopherol and selenium supplements in preventing lupinosis-associated myopathy in sheep

Objective To compare the effectiveness of combined selenium and α-tocopherol acetate treatments in preventing lupinosis-associated myopathy in sheep.
Design Measurement of plasma muscle and liver enzymes, and histopathological examination of muscle and liver in the treatment groups.
Procedure The treatments were: subcutaneous injections of selenomethionine and vitamin E (sc(SeM+E)) , an intraruminal selenium pellet and oral doses of vitamin E and intramuscular injections of selenomethionine combined with either oral doses of vitamin E (imSeM+orE) or intramuscular injections of vitamin E in an oily carrier. Another group received no supplements, while a control group was given selenium pellets on day 0 and fortnightly oral doses of vitamin E from day 0 to 72. To produce lupinosis-associated myopathy, the sheep were fed a diet low in vitamin E and given repeated injections of a crude extract of Diaphorthe toxica. Groups sc(SeM+E) and imSeM+orE were stressed by dosing with protected polyunsaturated fatty acids from day 56 onwards.
Results Lupinosis-associated myopathy was induced in all unsupplemented sheep. In these sheep the storage of Se increased and that of vitamin E decreased. The subcutaneous treatment was highly effective in preventing lupinosis-associated myopathy and also produced the highest vitamin E concentrations in plasma and liver. Supplemental vitamin E was more efficacious than supplemental Se. Concentrations of vitamin E in the livers of sheep given intramuscular vitamin E were higher than expected based on plasma concentrations. Oral doses of vitamin E proved the least effective method of increasing concentrations in liver. Lupinosis did not affect Se concentrations in liver or muscle.
Conclusion The sc(SeM+E) treatment is highly effective in preventing lupinosis-associated myopathy but needs to be further assessed when selenium and vitamin E are both limiting in the diet.