Blastocyst Evaluation by Means of Differential Staining: a Practical Approach

Techniques for in vitro production of embryos have been developed world-wide in different species, with promising results in human and ruminants. Thousands of human IVF-babies have been born during the last 20 years and thousands of in vitro -produced calves have been born since the late 1980s. With current methods for bovine in vitro fertilization, about 30–40% of in vitro -fertilized bovine oocytes develop further to the blastocyst stage and can be used for transfer. A proper evaluation of blastocyst quality remains however, an important challenge for every researcher involved in embryology and for every clinician who wants to select the best embryos for transfer. This review attempts to summarize the different methods available for estimation of blastocyst quality with a special emphasis upon differential staining.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

Dietary crude protein concentration does not affect the leucine requirement of growing dogs

The objective of the present study was to examine the interaction between graded levels of leucine and dietary crude protein. Dose–response curves were generated using four 3 × 3 Latin squares (two dogs/square). Each square represented one of two concentrations of crude protein (140 or 280 g/kg diet) and one of two combinations of three concentrations of leucine (5.0, 7.0 and 9.0 g/kg diet or 9.0, 11 and 13 g/kg diet). An additional experiment was performed by feeding crude protein at 210 g/kg diet with either 7.0 or 11 g leucine/kg diet. Weight gain, food intake, nitrogen retention, plasma albumin and plasma amino acids were measured. The requirement was determined to be the minimum leucine concentration required to maximize weight gain and nitrogen retention. For 8–14-week-old male Beagle dogs, 140 g crude protein/kg diet in a diet containing 18 kJ metabolizable energy/g does not appear to support maximal growth. The leucine requirement was not affected by doubling the dietary crude protein level from 140 to 280 g/kg diet. From these results, the leucine requirement of 8–14-week-old Beagle dogs appears to be 11 g leucine/kg diet independent of the level of dietary crude protein, whereas dogs over 14 weeks require only 7 g leucine/kg diet for maximal nitrogen retention.     

Risk factors for pet evacuation failure after a slow-onset disaster.

OBJECTIVE: To determine risk factors for pet evacuation failure during a flood. DESIGN: Cross-sectional survey. SAMPLE POPULATION: 203 pet-owning households in a flooded region. PROCEDURES: Persons under evacuation notice because of a flood were interviewed by use of a random telephone survey. RESULTS: 102 households evacuated with their pets, whereas 101 households evacuated without their pets. Low pet attachment and commitment scores were significantly associated with a greater chance of pet evacuation failure. Risk of pet evacuation failure and lower attachment and commitment scores were also associated with pet management practices prior to the disaster, such as dogs being kept outdoors most of the time or owners not having carriers for their cats. More than 90% of owners made housing arrangements for their pets without assistance. CONCLUSIONS AND CLINICAL RELEVANCE: Predictors of pet evacuation failure are usually present before a disaster strikes and are potentially modifiable. Mitigation of pet evacuation failure should focus on activities that reinforce responsible pet ownership and strengthen the human-animal bond, including socializing dogs, attending dog behavior training classes, transporting cats in nondisaster times, and seeking regular preventive veterinary care. Most pet owners are self-reliant in disasters, and this behavior should be encouraged.     

Protection of turbot, Scophthalmus maximus (L.), and rainbow trout, Oncorhynchus mykiss (Richardson), against vibriosis using two different vaccines

Abstact. The potency of a whole-cell bacterin (WCB) and a toxoid enriched whole-cell vaccine (WCEB) administered intraperitoncally into rainbow trout, Oncorhynchus mykiss (Richardson), were compared. The most effective vaccine was further evaluated by bathing turbot, Scophthalmus maximus (L.). These vaccines were composed of three strains of V. anguillarun , of the serotypes 01 and 02. Both vaccines conferred the highest protection against strains of serotype 01 within 4 weeks. With the toxoid enriched vaccine giving the best results (77 RPS). When trout were revaccinated after 7 weeks with this vaccine, good protection was achieved against strains of serotypes 01 and 02. Interestingly, when the WCEB was administered by bath to turbot, acceptable levels of protection against strains of both serotypes were obtained after 4 weeks of immunization.     

Hepatic Lobe Torsion as a Cause of Colic in a Horse

A 14-year-old Arabian gelding was examined for colic. An exploratory celiotomy was subsequently performed and the left lobe of the liver was found to be twisted. The lobe was resected using a TA-90 surgical stapling instrument. Histologic examination of the resected liver indicated portal vein and sinusoid dilation and congestion with blood. There were focal areas of necrosis and bacterial cocci and rods throughout the section. The histologic findings were consistent with hepatic lobe torsion. After surgery, the horse was treated with broad spectrum antibiotics, anti-inflammatory drugs, heparin, and intravenous fluids. The horse recovered without complications, although serum liver enzymes remained elevated for more than 1 week after surgery. Seven months after surgery the horse showed no adverse affects from the disease.     

Pharmacokinetics of midazolam administered concurrently with ketamine after intravenous bolus or infusion in dogs

Brown, S.A., Jacobson, J.D., Hartsfield, S.M. Pharmacokinetics of midazolam administered concurrently with ketamine after intravenous bolus or infusion in dogs. J. vet. Pharmacol. Therap. 16 , 419–425. Midazolam, a water-soluble benzodiazepine tranquilizer, has been considered by some veterinary anaesthesiologists to be suitable as a combination anaesthetic agent when administered concurrently with ketamine because of its water solubility and miscibility with ketamine. However, the pharmacokinetics of midazolam have not been extensively described in the dog. Twelve clinically healthy mixed breed dogs (22.2–33.4 kg) were divided into two groups at random and were administered ketamine (10 mg/kg) and midazolam (0.5 mg/kg) either as an intravenous bolus over 30 s (group 1) or as an i.v. infusion in 0.9% NaCl (2 ml/kg) over 15 min. Blood samples were obtained immediately before the drugs were injected and periodically for 6 h afterwards. Serum concentrations were determined using gas chromatography with electron-capture detection. Serum concentrations were best described using a two-compartment open model and indicated a t_½α of 1.8 min and t_½β.p of 27.8 min after i.v. bolus, and t_½α f 1–35 min and t_½β of 31.6 min after i.v. infusion. The calculated pharmacokinetic coefficient B was significantly smaller after i.v. infusion (429 ± 244 ng/ml) than after i.v. bolus (888 ± 130 ng/ml, P = 0.004). Furthermore, AUC was significantly smaller after i.v. infusion (29 800 ±6120 ng/h/ml) than after i.v. bolus (42 500 ± 8460 ng/h/ml, P < 0.05), resulting in a larger Cl_B after i.v. infusion (17.4 ± 4.00 ml/min/kg than after i.v. bolus (12.1 ± 2.24 ml/min/kg, P < 0.05). No other pharmacokinetic value was significantly affected by rate of intravenous administration.     

Lack of effect of recombinant bovine interferon alpha I1 in the treatment of experimentally-induced bovine warts.

Fifteen four-month old calves were inoculated, on five scarified sites on each side of the neck, with a suspension of ground wart tissue from a steer naturally infected with bovine papilloma virus 1. Warts started to appear about one month postinfection and were measurable in ten calves two months postinfection, when the trial started. After stratification on the size of the warts, all fifteen calves were allocated randomly to one of the following treatment groups: twice weekly intramuscular injections of 5 mg recombinant bovine interferon alpha I1 (rBoIFN alpha I1), weekly injection of 5 mg of rBoIFN alpha I1 or placebo, for three weeks. The biggest wart on each calf at the beginning of the trial was measured and photographs of all warts were taken weekly for five weeks. An analysis of covariance on the log of the volumes of warts during the five weeks of the trial showed a significant difference between groups (p = 0.026). Warts in treated groups tended to grow faster than in the placebo group.