The Effect of Oxidative Stress on Thawed Bulk‐Sorted Red Deer Sperm

The aims of this study were to assess the effects of the sex‐sorting process on post‐thaw sperm quality as well as on induced oxidative stress damage (H₂O₂ 0 mm  = H000; H₂O₂ 50 mm  = H050; H₂O₂ 100 mm  = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non‐sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H₂O₂ increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm‐sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non‐sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.     

Visualization of three-dimensional chaos

Most chaotic mixing experiments have been restricted to two-dimensional, time-periodic flows, and this has shaped advances in theory as well. A prototypical, bounded, three-dimensional flow with a moderate Reynolds number is presented; this system lends itself to detailed experimental observation and allows for high-precision computational inspection. The flow structure, captured by means of cuts with a laser sheet (experimental Poincare section), was visualized with the use of continuously injected fluorescent dye streams and revealed detailed chaotic structures with high-period islands.     

Hypotensive and vasorelaxant effects of ethanolic extract from Jatropha gossypiifolia L. in rats

The present work was carried out to examine the hypotensive effects of ethanolic extract (EE) from Jatropha gossypiifolia L. The oral administration of EE (125 or 250 mg kg(-1) day(-1)) caused a significant and dose-dependent reduction of the systolic blood pressure. The concentration-response curves to norepinephrine (NE) or Ca(2+) were non-parallelly shifted to the right and the maximum contractile responses were concentration dependently depressed by EE (0.1 or 0.5 mg/ml) in endothelium-deprived mesenteric artery. The cumulative additions of EE (0.1-30 mg/ml) caused a concentration-dependent relaxant response in endothelium-deprived mesenteric artery precontracted with NE or Ca(2+). In conclusion, our results have shown that the EE from J. gossypiifolia L. can elicit hypotension, by oral via, in conscious normotensive rats and vasorelaxant activity on rat mesenteric rings precontracted with NE or Ca(2+).     

Activation of goat primordial follicles in vitro: Influence of alginate and ovarian tissue

The present study aimed to evaluate the effect of three culture systems on caprine primordial follicle activation in vitro: follicles cultured either in the isolated form within alginate (Isolated follicles + Alginate treatment), or enclosed in ovarian tissue (in situ), with or without alginate (Fragment + Alginate, and Fragment alone treatments, respectively). After culture, the Isolated follicles + Alginate treatment presented a percentage of morphologically normal follicles (MNF) similar to both the non-cultured control and the Fragment Alone treatments. Nevertheless, Fragment + Alginate treatment showed a significant reduction in the number of MNF when compared to the other treatments. Regarding follicle development, our results showed that regardless of the alginate, the presence of ovarian tissue limited primordial follicle activation during in vitro culture. Remarkably, the Isolated primordial follicle + Alginate treatment was the only one that significantly promoted follicle activation and increased both follicle and oocyte diameters during IVFC, pointing out a higher cell proliferation. In conclusion, the presence of ovarian tissue with or without alginate limited follicle development (activation) after culture. Nevertheless, when primordial follicles were isolated and encapsulated in alginate they presented suitable survival rates, higher rates of follicle activation and continued to grow throughout the culture period.     

Tuberculosis in a New Zealand (Hooker's) sea lion

Extract

Tuberculosis has been described in seven species of pinnipeds. Based on host preference, phenotype and genetic and antigenic testing, the causal bacterium has been classified as a distinct species, Mycobacterium pinnipedii, within the M. tuberculosis complex. An adult male New Zealand sea lion (Phocarctos hookeri) was found dead on the Otago coastline in mid 2005. Gross necropsy revealed multiple caseous foci throughout the lungs, subcutaneous and thoracic lymph nodes, and mesenteric lymph nodes. Histopathological examination showed granulomatous lesions containing numerous intra-lesional acid fast organisms. The organism was confirmed as being a member of the M. tuberculosis complex, based on restriction enzyme analysis. Spoligotyping is pending. This is the first confirmed case of mycobacterial disease in a New Zealand sea lion.     

The effect of four anesthetic protocols on the spleen in dogs

Splenic enlargement following administration of barbiturates has been well described in dogs; other agents have not been investigated. This study aimed to compare the effects of four anesthetic protocols on splenic size.
Twenty-four fasted Beagle dogs scheduled for laparotomy were allocated to one of the four groups. Group 1: acepromazine and butorphanol followed by induction with thiopental; Group 2: acepromazine and butorphanol followed by induction with propofol; Group 3: medetomidine and butorphanol followed by induction with propofol; Group 4: medetomidine and butorphanol followed by induction with ketamine and diazepam. Anesthesia was maintained with halothane in oxygen, intravenous fluids were administered. Splenic length, width and height were measured once when the abdomen was opened and again just prior to closure. Spleens were also traced, the image was digitized, and the area was calculated. PCV and total solids were measured before and after pre-medication, after induction, and each time the spleen was measured. Data were analyzed using a Repeated Measures anova with splenic variables indexed by body surface area and dose of induction agent as a covariate.
Area and width of the spleens were less in the dogs of Groups 2 and 3 than in those of the other groups. Splenic area and length did not change significantly during surgery. Dosage of propofol was not significantly different between Groups 2 and 3. Baseline PCV was not significantly different among groups and decreased significantly in all dogs, but at different times. In Groups 1 and 2, the decrease occurred after pre-medication, in Group 3 at induction, and in Group 4 during surgery. A significant decrease in TS occurred in all groups during surgery.
We concluded that the use of propofol resulted in smaller spleen size during surgery than that following the use of thiopental. Multiple factors influenced the PCV.     

Evaluation of intraperitoneal and subcutaneous lidocaine and bupivacaine for analgesia following ovariohysterectomy in the dog

The role of ketamine (K) in pain management is controversial. It is reported to provide visceral analgesia in cats. This study aimed to assess its somatic actions using a thermal threshold (TT) model. Six cats (four spayed females, two castrated males, 4.3–7.2 kg) participated in the study. The day before each study, the thorax of each of the cats was shaved and a cephalic catheter was placed. TT was measured using a device specifically developed for cats. A heater element and temperature sensor housed in a small probe were held against the thorax of the cats with an elastic band and pressure bladder to assure consistent contact. The skin temperature was recorded before each test, then the heater was activated. When the cat responded by flinching, turning, or jumping, the stimulus was terminated and the threshold temperature was recorded. Treatments were 2 mg kg^?1 of K (10 mg mL^?1), or 0.2 mL kg^?1 of saline (S) IV, given in a randomized cross‐over design with at least 1 week between treatments. The investigator was blinded to the treatment. TT was measured thrice before treatment (baseline threshold) at 15 minutes, then every 30 minutes for 8 hours and once at 24 hours after injection. Data were analyzed using a four‐factor anova . Cats were sedated for 45 minutes following K treatment. There was no difference in baseline TT between treatments (K = 41.9 ± 1.7 °C, S = 41.0 ± 1.45 °C), and no change in TT at any time in the S group. TT increased significantly at 15 and 30 minutes after K, then decreased below baseline values between 210 and 390 minutes, with a nadir of 38.8 ± ± 1.05 °C at 390 minutes. During this time period, cats exhibited normal activity, but responses to thermal stimuli were exaggerated. This study suggested that K caused a delayed onset hyperalgesia in cats.     

Differentiation of Australian isolates of Mycobacterium paratuberculosis using pulsed-field gel electrophoresis

Objectives To examine strain variation amongst Australian isolates of Mycobacterium paratuberculosis .
Design Pulsed field gel electrophoresis was optimised for differentiation of M paratuberculosis strains, and this typing technique was then applied to a collection of Australian isolates.
Procedure DNAs from 35 Australian isolates of M para-tuberculosis and a UK reference strain were digested with one or other of three restriction endonucleases. The banding patterns obtained after pulsed field gel electrophoresis of the DNA fragments were compared.
Results The Australian isolates were divided into two groups on the basis of their DNA banding pattern. Both were different from the UK reference strain. Seven isolates from cattle in Victoria and the Northern Territory had the same pattern as five isolates from alpacas in Victoria and Western Australia. Another 20 isolates from cattle in Victoria, Western Australia and the Northern Territory had the same pattern as isolates from two sheep and a goat in New South Wales.
Conclusion Pulsed field gel electrophoresis was a useful tool for strain typing of M paratuberculosis , and could be used to study the transmission of strains in Australia.     

Characterization of two Acanthoscelides obtectus alpha-amylases and their inactivation by wheat inhibitors

Wheat alpha-amylase inhibitors represent an important tool in engineering crop plants against bean bruchids. Because Acanthoscelides obtectus is a devastating storage bean insect-pest, we attempted to purify and characterize its gut alpha-amylases, to study their interaction with active proteinaceous inhibitors. Two digestives alpha-amylases (AoA1 and AoA2) were purified from gut larvae, showing molecular masses of 30 and 45 kDa for each one, respectively. The stoichiometry interaction between these alpha-amylases with two wheat inhibitors (0.19 and 0.53) showed a binding complex of 1:1 enzyme:inhibitor. In vivo activities of these inhibitors against A. obtectus were also evaluated using a rich ammonium sulfate inhibitor fraction (F(20)(-)(40)) and purified inhibitors after reversed phase high-performance liquid chromatography columns. Incorporation of three different inhibitor concentrations (0.25, 0.5, and 1.0% w/w) into artificial seeds showed that addition of the purified 0.19 inhibitor at the highest concentration (1.0%) reduced the larval weight by 80%. Similar data were observed when 0.53 inhibitor was incorporated at 0.5%. When the concentration of purified 0.53 was enhanced to 1.0%, no larvae or adult emergence were observed. Our data suggest that these alpha-amylase inhibitors present great potential for use in Phaseolus genetic improvement programs.