Effects of Retinoids on the In Vitro Development of Capra Hircus Embryos to Blastocysts in Two Different Culture Systems

The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre‐implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus‐oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus‐oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39°C in an atmosphere of 5% (v/v) CO₂ in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39°C in a humidified atmosphere of 5% (v/v) CO₂, 5% (v/v) O₂ and 90% (v/v) N₂. In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO₂ in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 μg/ml RT and 0.5 μm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre‐implantation development of goat embryos and can be used to enhance in vitro embryo production.     

Characterization of structure and function of the mouse retina using pattern electroretinography, pupil light reflex, and optical coherence tomography

Objective To perform in vivo analysis of retinal functional and structural parameters in healthy mouse eyes. Animal Studied Adult C57BL/6 male mice (n?=?37). Procedures Retinal function was evaluated using pattern electroretinography (pERG) and the chromatic pupil light reflex (cPLR). Structural properties of the retina and nerve fiber layer (NFL) were evaluated using spectral-domain optical coherence tomography (SD-OCT). Results The average pERG amplitudes were found to be 11.2?±?0.7?μV (P50-N95, mean?±?SEM), with an implicit time for P50-N95 interval of 90.4?±?5.4?ms. Total retinal thickness was 229.5?±?1.7?μm (mean?±?SEM) in the area centralis region. The thickness of the retinal nerve fiber layer (mean?±?SEM) using a circular peripapillary retinal scan centered on the optic nerve was 46.7?±?0.9?μm (temporal), 46.1?±?0.9?μm (superior), 45.8?±?0.9?μm (nasal), and 48.4?±?1?μm (inferior). The baseline pupil diameter was 2.1?±?0.05?mm in darkness, and 1.1?±?0.05 and 0.56?±?0.03?mm after stimulation with red (630?nm, luminance 200?kcd/m(2) ) or blue (480?nm, luminance 200?kcd/m(2) ) light illumination, respectively. Conclusions Pattern electroretinography, cPLR and SD-OCT analysis are reproducible techniques, which can provide important information about retinal and optic nerve function and structure in mice.     

Congenital portosystemic shunts in Maltese and Australian Cattle Dogs

SUMMARY Congenital portosystemic shunts were definitively diagnosed in 62 dogs over a period of 15 years. Maltese and Australian Cattle Dogs were significantly over-represented, accounting for 14 and 13 cases, respectively. Maltese invariably had a single extrahepatic shunt derived from the left gastric or gastrosplenic vein, whereas Cattle Dogs usually had large intrahepatic shunts involving the right liver lobes. The clinical syndromes resulting from anomalous portosystemic communications were indistinguishable in the 2 breeds. Fasting blood ammonia concentration was elevated in 20 of 22 dogs tested, providing a minimally invasive and effective means of diagnosis. Complete or partial shunt attenuation was performed successfully in all 9 Maltese and in 2 of 6 Cattle Dogs in which it was attempted.     

Canine malignant hyperthermia susceptibility: erythrocytic defects--osmotic fragility, glucose-6-phosphate dehydrogenase deficiency and abnormal Ca2+ homeostasis.

Two dogs were diagnosed as malignant hyperthermia susceptible based on increased susceptibility (P less than 0.001) of biopsied muscle to caffeine-induced contracture. Erythrocytes from malignant hyperthermia and normal dogs were then examined for an antioxidant system deficiency. Values for serum muscle enzymes, reticulocytes and corpuscular hemoglobin were mildly elevated. Osmotic fragility was increased: hemolysis occurred at a NaCl concentration 10 mM higher than for normal dogs (P less than 0.001). A 35% glucose-6-phosphate dehydrogenase deficiency (P less than 0.001) with a 40% compensatory increase (P less than 0.01) in 6-phosphogluconate dehydrogenase activity was found. The membrane Ca2+-activated ATPase activity was abnormal: 100% increased with a 40% decreased Arrhenius activation energy (P less than 0.005) and increased thermostability. A 40% increased intracellular accumulation of total Ca2+ occurred in response to in vitro energy depletion in erythrocytes from one malignant hyperthermia dog (P less than 0.01). The multifactorial pattern of inheritance and the broad spectrum of malignant hyperthermia susceptibility are proposed to result from an antioxidant system deficit unmasking or aggravating an intrinsic muscle membrane anomaly. An individual from a family with a history of malignant hyperthermia or unexplained anesthetic death should be considered malignant hyperthermia susceptible if erythrocyte osmotic fragility is abnormal and there is a mild, unexplained elevation in serum creatine kinase.     

响应面法优化姬松茸液体发酵培养基中红糖和酵母膏浓度

运用响应面分析法优化姬松茸液体发酵培养基中红糖和酵母膏浓度.结果表明,红糖浓度为20.24g/L、酵母膏浓度为2.055g/L时,菌丝体生物量最高,达2.279g/L;红糖浓度为20.8g/L、酵母膏浓度为2.23g/L时,胞外多糖产量最高,达0.749g/L.     

Sonographic features of drug-induced splenic congestion

Drug-induced splenic congestion has been reported in dogs secondary to barbiturate administration. This research attempted to verify and quantify size changes associated with drug-induced splenic congestion in dogs. Transverse plane ultrasound images of the spleen in normal dogs were collected to determine the maximum diameter in the minimum dimension prior to, and 15 min after, administration of acepromazine, thiopental, or propofol. Significant splenic enlargement was seen after administration of acepromazine (P<0.01) and thiopental (P=0.02), but no enlargement was seen after administration of propofol. Significantly increased attenuation (P<0.01) and a trend of increased backscatter (P=0.09) were measured after administration of acepromazine. These results indicate that measurable splenomegaly occurs after acepromazine and thiopental administration. This represents the first report of a condition causing measurable diffuse increased attenuation in the spleen. Propofol does not cause measurable splenic enlargement under the conditions of this research.     

CONTRAST HARMONIC ULTRASOUND OF SPONTANEOUS LIVER NODULES IN 32 DOGS

Thirty-two dogs with spontaneous hepatic nodules were given intravenous ultrasound contrast medium (Definity or Sonovue) and imaged with contrast harmonic software on a conventional ultrasound machine system. Digital video images were initially reviewed to describe the perfusion pattern of malignant nodules. The images were reviewed again to test this pattern against all individual nodules. Subjectively, there was improved conspicuity of malignant nodules after contrast enhancement compared with conventional imaging and increased numbers of malignant nodules were often noted. There was decreased conspicuity of benign nodules and no additional nodules were seen after contrast enhancement. There was a highly significant (P < 0.0001) association of malignancy with a hypoechoic nodule at surrounding normal liver peak contrast enhancement. Benign nodules were isoechoic to the surrounding normal liver at peak contrast enhancement. Only one benign nodule (hepatoma) had regions of hypoechogenicity compared with the surrounding normal liver at peak liver contrast enhancement. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were highly significant (P < 0.0001) (100%, 94.1%, 93.8%, 100%, and 96.9%, respectively). No complications or morbidity was noted throughout the course of the study. Contrast harmonic ultrasound appears to be accurate at discriminating between naturally occurring benign and malignant nodules in the liver of dogs.     

Successful Low Dose Insemination of Flow Cytometrically Sorted Ram Spermatozoa in Sheep

The fertility of ram spermatozoa that had undergone flow cytometric sorting (MoFlo SX) and cryopreservation was assessed after low-dose insemination of synchronized Merino ewes. Oestrus was synchronized with progestagen-impregnated pessaries, PMSG and GnRH treatment. Ewes (n = 360) were inseminated with 1 x 10(6), 5 x 10(6) or 15 x 10(6) motile sorted frozen-thawed (S(1), S(5), or S(15) respectively) or non-sorted frozen-thawed (C(1), C(5) or C(15) respectively) spermatozoa from three rams. An additional group of ewes were inseminated with 50 x 10(6) motile non-sorted frozen-thawed spermatozoa (C(50)) to provide a commercial dose control. The percentage of ewes lambing after insemination was similar for C(50) (24/38, 63.2%), C(15) (37/54, 68.5%), S(15) (38/57, 66.7%), S(5) (37/56, 66.1%) and S(1) (32/52, 61.5%) groups (p > 0.05), but lower for C(5) (19/48, 39.6%) and C(1) (19/55, 34.5%) treatments (p < 0.05). This study demonstrates sorted ram spermatozoa are equally fertile to non-sorted spermatozoa even when inseminated at 2% of the dose. Furthermore, at very low artificial insemination doses (1 or 5 million motile) the fertility of sorted ram spermatozoa is superior to non-sorted spermatozoa inseminated in equal numbers. These results have significance for the future commercialization of sex-preselection technology in sheep as a reduction in the minimum effective sperm number will allow a corresponding decrease in the associated cost per dose.     

仔猪腹泻病的鉴别诊断及综合防治

对仔猪腹泻病进行了摸索、研究和总结,根据不同日龄和不同临床表现进行鉴别诊断,同时可采取组织病理学、微生物学方法进一步确诊,并采取针对性防治措施,才能有效地控制仔猪腹泻病.