Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

玉米杂交种辽单27号选育报告

1　亲本来源辽单 2 7号 (原名辽 940 4 )为辽宁省农业科学院作物研究所以自选系辽 2 30 9为母本 ,引入系丹340为父本杂交组配而成。辽 2 30 9自选系是 1 990年以美国先锋杂交种7861 3(ｏｐ)为基础材料连续自交多代选择育成。辽 2 30 9一般配合力高 ,高抗大、小斑病和丝黑穗病 ,抗病毒病和茎基腐病 ,秆硬抗倒 ,株形紧凑 ,活秆成熟 ,产量高。2　辽单 2 7号选育经过1 992年冬在海南岛测配时用丹 340 (是当时测验种之一 )配制成辽 2 30 9×丹 340杂交组合。1 993～ 1 994年进行院内品比试验。 1 995年参加辽宁省区域试验预备试验 ,1 996～ 1 997…     

Fiber characteristics of pork muscle exhibiting different levels of drip loss

The present study aimed to investigate the fiber characteristics of pork muscle exhibiting different levels of drip loss during storage. The samples were taken from Longissimus dorsi muscle to evaluate drip loss (n = 100). Fifteen muscles per group (low and high drip loss) were selected to evaluate the histological characteristics at 0 and 72 h of storage time. The statistical analysis revealed that a high drip loss group had greater endomysium thickness than a low drip loss group at 0 h of storage time (P < 0.05). There were no significant differences in total number of fibers, fiber diameter, fiber area and perimysium thickness at 0 h of storage time (P > 0.05). At 72 h of storage time, a high drip loss was evident in higher total number of fibers (P < 0.01), but smaller fiber diameter (P < 0.05), fiber area (P < 0.01) and endomysium thickness (P < 0.01) than a low drip loss group. There was no significant difference in perimysium thickness (P > 0.05). In conclusion, drip loss might be affected by muscle structural characteristics during storage.     

Evaluation of bee venom and hyaluronic acid in the intra-articular treatment of osteoarthritis in an experimental rabbit model

The aim of this study was to investigate bee venom and hyaluronic acid in the intra-articular treatment of osteoarthritis in an experimental rabbit model. Forty-five rabbits were used and they were randomly divided into three groups (BVI, BVII, and HA) and each group was divided to two subgroups to evaluate the radiologic, magnetic resonance imaging, histopathologic, and biochemical evaluation in post treatment second week (a) and twelfth week (b). Radiologically, a significant difference was observed in the HA group (P<0.05). The MRI evaluation of at any time in group BVI(b) was found to be different. No significant differences were seen between the groups, biochemically. Histopathologically, cellularity, and orthochromasia was evident with Safranin-O in the BVI(b) and BVII(a); adhesions were seen in the BVII(a) group and clustering of chondrocyte in the HA(b) group were found to be different. Consequently, intra-articular application of HA and BV for experimental model of osteoarthritis has no significant influence upon recovery after therapy.     

Magnetic resonance imaging findings in dogs with thoracolumbar intervertebral disk disease: 69 cases (1997-2005)

OBJECTIVE: To determine magnetic resonance imaging (MRI) abnormalities in dogs with intervertebral disk disease (IVDD) and develop a classification scheme for IVDD in dogs based on MRI findings. DESIGN: Retrospective case series. ANIMALS: 69 dogs. PROCEDURE: Medical records of dogs admitted because of thoracolumbar IVDD in which MRI of T9 through L7 had been performed were reviewed. RESULTS: A total of 759 intervertebral disk spaces were examined. Of these, 342 (45.1%) were classified as having a normal MRI appearance; the remaining 417 (54.9%) had various types of IVDD. Disk degeneration was identified in 276 disk spaces in 56 dogs, bulging of the intervertebral disk was identified in 37 disk spaces in 24 dogs, disk protrusion was identified in 54 disk spaces in 32 dogs, and disk extrusion was identified in 50 disk spaces in 48 dogs. Cartilage endplate changes were identified in 35 vertebrae in 17 dogs, and increased signal intensity of the spinal cord was identified in 21 dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Four types of IVDD (disk degeneration, bulging of the intervertebral disk, disk protrusion, and disk extrusion) were identified on the basis of MRI findings in dogs with thoracolumbar IVDD. We recommend that a standardized nomenclature be adopted for the various types of thoracolumbar IVDD in dogs.     

Downstream fish migration from two regulated lakes monitored by hydroacoustics

The migration of fish from two large northern Finnish lakes to their outflowing rivers was studied by echosounding and exploratory fishing. Both lakes are regulated for hydroelectric purposes. In both rivers, two sonar stations with stationary up- and down-looking transducers were used to collect data for one year. The fish migration rate in the River Oulujoki was greater than in the River Paatsjoki. In the River Oulujoki, the fish migrated mainly downstream and in the River Paatsjoki both down- and upstream. In the River Paatsjoki, larger fish showed active migration in the spring and autumn, whereas in the River Oulujoki the increase in the migration occurred simultaneously in all size groups. The different species composition and the different nature of the lakes, together with the different regulation practices, were considered to be responsible for the varying migration patterns. It was concluded that no barriers to fish migration should be built on these rivers.     

Factors affecting the natural transmission of bovine leukaemia virus infection in Queensland dairy herds

Natural transmission of bovine leukaemia virus (BLV) infection in south-eastern Queensland dairy herds was slow in 2 herds with a low to moderate (13 to 22%) prevalence of infection. Infection spread much more rapidly in a herd that had a higher prevalence (42%) when first tested. In a 13 month study of this herd, the cumulative incidence of infection was 24%. In one herd new infections were confined almost entirely to calves of uninfected dams. Following the end of feeding bulk milk to calves, a common practice in dairy herds, no more calves in this herd became infected. In laboratory experiments, neither prolonged housing of susceptible calves with infected cattle, consumption of drinking water contaminated with infected blood, nor inoculation of sheep with saliva from infected cattle resulted in transmission of BLV infection. Sheep were infected by subcutaneous inoculation of a suspension of purified lymphocytes from an infected heifer. The minimum infective dose was 10(3) lymphocytes, equivalent to the number of lymphocytes in approximately 0.1 microliter blood. Thus, procedures involving the transfer of a very small volume of blood from animal-to-animal have the potential to transmit infection.     

Introducing the Karayaka sheep breed with its traits and influencing factors

This study was planned to express the detailed traits of Karayaka sheep reared in Tokat province, also influencing factors on the traits were evaluated. While year effect was significant on 8 weeks weight, scanning weight, fat depth and muscle area, all the evaluated traits including birth weight and muscle depth were significantly affected by season (p < 0.05). Dam age was significant on birth weight (p < 0.05). Birth weight and 8 week weight were significantly affected by sex and birth type. Sex effect has been appeared on muscle depth while birth type effect has been detected on scanning weight (p < 0.05). While the strongest phenotypic correlation coefficient was detected between muscle depth and muscle area, the weakest ones were found between birth weight and scanning traits.     

Effect of Dietary N‐Carbamylglutamate Levels on Reproductive Performance of Gilts

The effect of dietary N‐carbamylglutamate (NCG) supplementation during the entire gestation on reproductive performance of gilts was determined. At the initial day of gestation, forty‐five cross‐bred (Landrace × Large white) gilts were randomly assigned to five groups receiving a basal diet (control group) and basal diet supplemented with 0.05%, 0.10%, 0.15% and 0.20% NCG until parturition, respectively. At parturition, total litter size, live litter size and rate of stillbirth were not markedly affected by NCG supplementation. However, gilts in 0.05% NCG‐supplemented group had more pigs born alive than gilts in control group (+1.11 pigs, p = 0.12), and live litter weight was increased (+12.13–19.17%, p < 0.05) in 0.05%, 0.10% and 0.15% NCG‐supplemented groups relative to control group. And also, average birthweight of piglets born alive was higher (+6.57%, p < 0.05) in 0.05% NCG‐supplemented group than in control group. Furthermore, on days 30, 60, 90 and 110 of gestation, concentrations of arginine and ornithine in plasma were higher (p < 0.05) in 0.05%, 0.10%, 0.15% and 0.20% NCG‐supplemented groups than in control group, respectively. In addition, the chorioallantois gene expression of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor A (VEGF‐A), placental growth factor (PLGF) and angiopoietin‐2 (ANG‐2) was all increased (p < 0.05) in 0.05%, 0.10% and 0.15% NCG‐supplemented groups than in control group. In conclusion, dietary supplementation with 500 mg/kg NCG during the entire gestation significantly improves pregnancy outcomes in gilts, which may be associated with the improved concentrations of arginine in plasma and placental angiogenic factors gene expression of gilts.     

Effects of Gonadotropins on In Vitro Maturation and of Electrical Stimulation on Parthenogenesis of Canine Oocytes

The objective of this study was to determine the effects of gonadotropins on in vitro maturation (IVM) and electrical stimulation on the parthenogenesis of canine oocytes. In experiment I, cumulus oocyte complexes were collected from ovaries at a random phase of the oestrus cycle and cultured on maturation medium treated with hCG or eCG for 48 or 72 h. There were no significant differences in the effects on the metaphase II (MII) rate between the hCG and eCG treatment groups over 48 h (5.4% vs 5.5%). The MII rate in the co-treatment group of hCG and eCG for 48 h was higher than in each hormone treated group (15.5%, p < 0.05). In experiment 2, the parthenogenetic effect on oocyte development, at various electrical field strengths (1.0, 1.5, 2.0 kV/cm DC) for 60 or 80 μs with a single DC pulse after IVM on the co-treatment of hCG and eCG, was examined. The rate of pronuclear formation (37.1%) in electrical activation at 1.5 kV/60 μs without cytochalasin B (CB) was higher than that of oocytes activated in the other groups (p < 0.05). However, we did not observe the cleavage stages. Also, CB did not influence parthenogenesis of canine oocytes. The results showed that the pronucleus formation rate, indicative of the parthenogenesis start point, could be increased by electrical stimulation. Therefore, these results can provide important data for the parthenogenesis of canine oocytes and suggest the probability of parthenogenesis in canines.