Cryopreservation and In Vitro culture of Preimplantation Embryos in Djungarian Hamster (Phodopus sungorus)

Although embryo cryobanking was applied to Syrian golden and to Campbell's hamsters, no attempt has been made at freezing embryos in Djungarian hamsters. Four‐cell stage embryos were flushed from the reproductive ducts of pregnant females before noon of the third‐day post coitum and frozen in 0.25‐ml straws according to standard procedures of slow cooling. A mixture of permeating (ethylene glycol) and non‐permeating (sucrose) cryoprotectants was used. The thawing was performed by incubating at RT for 40 s followed by 40 s in a water bath at 30.0°C. Most (66.7%) of the non‐frozen four‐cell embryos developed up to the morula stage in rat one‐cell embryo culture medium (R1ECM). The use of hamster embryo culture medium (HECM) yielded fewer morulas (18.2%) during the same 24‐h period of culture. The rate of embryo's surviving the freezing–thawing procedures, as estimated by light microscopy, was 60.7–68.8%. After 24‐h culturing in R1ECM, 64.7% of frozen–thawed four‐cell embryos developed and all of them reached the morula stage. Supplementation of R1ECM with GM‐CSF (2 ng/ml) improved the rate of Djungarian hamster frozen–thawed embryo development: 100% of the four‐cell stage embryos developed, 50% of them achieved the morula stage, and 50% developed even further and reached the blastocyst stage within 24 h of culturing. This study reports the world's first successful transfer of frozen–thawed Djungarian hamster embryos yielding term pups. Taken together, the results of this study demonstrate the possibility of applying some key reproductive technologies, that is, embryo freezing/cryopreservation and in vitro culture, to Djungarian hamsters.     

Cardiac arrhythmias associated with acute major haemorrhage during craniectomy in a dog

An estimated 43% of total blood volume was lost during craniectomy in a 32‐kg Labrador retriever. One episode of bradycardia, followed by ventricular tachycardia occurred during surgery when the rate of haemorrhage exceeded fluid replacement. Sinus rhythm was re‐established with intravenous lidocaine (1.25 mg kg^?1). Twenty‐four hours later, premature ventricular complexes appeared followed by episodes of ventricular tachycardia, some requiring lidocaine treatment. Myocardial hypoxia resulting either from hypovolaemia and/or air embolism (the right transverse venous sinus was damaged during surgery) may have caused the arrhythmias. Mild intracranial hypertension may have aggravated the problem.     

Effect of some substituted benzonitriles on photosynthetic activity of spinach and wheat in vivo and in vitro

The inhibitory effects of nine nitro and/or bromo-substituted benzonitrile compounds on the photosynthetic electron flow in isolated chloroplasts and on the in vivo CO₂ fixation of spinach (Spinacia oteracea L.) and wheat (Triticum aesticum L. cv. Bezoslaya) were investigated. Bromoxynil and 3-nitro-5-hromo-4-hydroxy-bcnzonitrile were the strongest and equally effective inhibitors of Ihe in vivo CO₂ fixation of spinach, hut in wheat the nitro-bromo-compound is ineffective and 3-nitro-benzonitrile is even more inhibitory than bromoxynil. None of the substances affected DCPIPH → methylviologen reduction. In the inhibition of the DCPIP reduction only the 3,5-disuhstituted 4-hydroxy-derivatives were effective. The fact that these compounds affect only the PS II reaction with both H₂O and DPC as electron donors suggests a site of inhibition on the reducing side of PS II, between Ihe PS II reaction centre and ihe DCPIP Site. It is suggested that in the inhibition of the DCPIP reduction only steric factors are important and the different electron configuration of the sterieally similar molecules may be involved only in the absorption and translocation processes of the compounds.     

Paraoxonase (PON) 1, 2 and 3 Expression in Granulosa Cells and PON1 Activity in Follicular Fluid of Dairy Cows

Normal metabolic activity in ovarian follicles may result in oxidative stress and damage to oocytes. The aim of this study was to evaluate expression of the natural anti‐oxidants paraoxonase (PON) 1, 2 and 3 in granulosa cells and PON1 activity in follicular fluid (FF) and plasma of dairy cows. For the first experiment, ovaries were collected from cows at slaughter, after which follicles were dissected and classified as oestrogen active (EAF) or atretic (ATF). Expression of PON1, PON2 and PON3 mRNA was evaluated in granulosa cells, and activity of PON1 was measured in FF. PON1 mRNA was undetectable in granulosa cells, PON2 mRNA expression was not different between follicle types, and PON3 mRNA tended to be higher in EAF (p = 0.11). The activity of PON1 in FF was higher (p = 0.01) for EAF (82.6 ± 8.0 kU/L) than ATF (53.9 ± 6.8 kU/L), as were high‐density lipoproteins (HDL), low‐density lipoproteins (LDL) and total cholesterol concentrations. In the second experiment, we aimed to compare plasma and FF PON1 activity in early lactation Holstein cows (n = 15) with pre‐ovulatory EAF. Activity of PON1 was twofold higher (p < 0.0001) in plasma (122.5 ± 11.1 kU/L) than in FF (61.4 ± 5.2 kU/L). Plasma concentrations were also higher (p < 0.0001) for HDL, LDL and total cholesterol when compared to FF. In conclusion, FF concentrations of PON1, HDL, LDL and total cholesterol were higher in healthy oestrogen active bovine follicles than in atretic follicles. PON1 was not expressed by granulosa cells indicating that high PON1 activity in bovine FF is apparently derived by transfer from blood in association with HDL.     

The Effects of Anesthesia on Laryngeal Function and Laryngeal/Pharyngeal Trauma in the Horse

Laryngeal paralysis in horses has been reported after inhalational anesthesia and can result in significant morbidity/mortality. The cause of the condition is unclear. The objective of this study was to examine the effects of a standardized anesthetic protocol on laryngeal function and laryngeal/pharyngeal trauma in the peri-anesthetic period in a prospective study. A 30- to 60-second digitalized video clip of laryngeal movement from a standardized endoscopic view was recorded at five time points: before sedation, post-sedation, post-induction, immediately after recovery to standing, and at 24 hours after recovery. A standardized anesthetic regimen was used in all cases. Video clips were randomized and evaluated by two blinded assessors. Each assessor scored each clip for laryngeal function and trauma using previously validated scoring systems. Agreement between assessors was calculated using the mean of the five time-specific weighted kappa statistics. Post-anesthesia laryngeal function and trauma scores were compared with initial scores using the Wilcoxon signed rank test with Bonferroni adjustment. Spearman's rank coefficient was used to assess correlation between trauma and function scores and between anesthetic duration and laryngeal function and trauma scores.There was no significant effect of anesthesia on laryngeal function. Trauma scores were not significantly higher after tracheal intubation. The trauma scoring system requires further validation. There was no correlation between higher trauma scores and laryngeal function or duration and laryngeal function or trauma.Further work is required to evaluate other variables that may affect laryngeal function after anesthesia, using a larger number of horses with varying degrees of laryngeal dysfunction.     

Pathogenicity of Vietnamese enterotoxigenic Escherichia coli strains in colostrum-deprived one-day-old piglets

Preweaning colibacillosis is a major cause of economic loss to the swine industry in Vietnam. The aim of this study was to examine the enteropathogenicity of representative enterotoxigenic Escherichia coli (ETEC) strains obtained during an earlier epidemiologic survey conducted in five provinces in North Vietnam. This included isolates belonging to serotype O8 that produced heat-stable and heat-labile enterotoxins but did not produce any of the recognized fimbriae (F4, F5, F6, F41, F18). In vitro hemagglutination (unique mannose-resistant hemagglutination activity with guinea pig, sheep, human, and chicken red blood cells at 37 degrees C, but not at 18 degrees C) and enterocyte brush border attachment assays suggested that the F- ETEC strains produced an unidentified colonization factor that promoted adherence to the intestinal epithelium. Colostrum-deprived 1-day-old piglets challenged with an F- strain (1-2 x 10(9) bacteria) developed acute watery diarrhea within 4 hours of inoculation and suffered up to 20% weight loss, with comparable severity to piglets challenged with conventional F4 and F5 strains. At necropsy, viable counts and histopathologic examination of intestinal sections demonstrated colonization of the duodenum, jejunum, and ileum by F4-positive strains. In comparison, the F- and F5-positive strains attached exclusively to the ileum. Transmission electron micrographs of negatively stained F- cells grown at 37 degrees C demonstrated the presence of fimbriae. These results confirm the presence of a potentially new pathogenic ETEC fimbrial type in piggeries in Vietnam, with a unique hemagglutination property and attachment characteristics similar to ETEC bearing F5 fimbriae.