Inactivation of the Wnt/β-catenin signaling contributes to the epithelial barrier dysfunction induced by sodium oxalate in canine renal epithelial cells

High oxalate consumption has been recognized as a risk factor for renal calcium oxalate stones in companion animals (dogs and cats). However, the cellular signaling involved in oxalate-induced dysfunction in renal tubular epithelial cells remains not fully elucidated. In this study, Mardin–Darby canine kidney (MDCK) cells, an epithelial cell line derived from canine kidney tubule, were tested for cell proliferation activity and barrier function after being exposed to sodium oxalate (NaOx). Further, the involvement of Wnt/β-catenin in NaOx-induced renal epithelial barrier dysfunction was evaluated. MDCK cells treated with NaOx exhibited reduction in cell proliferation and migration. Besides, NaOx exposure led to a decrease in transepithelial electrical resistance and an increase in paracellular permeability. The deleterious effects of NaOx on epithelial barrier function were related to the suppressed abundance of tight junction proteins including zonula occludens, occludin, and claudin-1. Of note, protein levels of β-catenin and phosphorylated (p)-β-catenin (Ser552) in MDCK cells were repressed by NaOx, indicating inhibitory effects on Wnt/β-catenin signaling. An inhibition of glycogen synthase kinase-3β (GSK-3β) by SB216763 enhanced the abundance of β-catenin and p-β-catenin (Ser552), and protected against epithelial barrier dysfunction in NaOx-treated MDCK cells. The results revealed a critical role of Wnt/β-catenin signaling in the epithelial barrier function of MDCK cells. Activation of Wnt/β-catenin signaling might be a potential therapeutic target for the treatment of oxalate-linked renal stones.     

Hyperglycaemia in transition dairy cows: Effects of lactational stage and conjugated linoleic acid supplementation on glucose metabolism and turnover

Supplementing conjugated linoleic acid ( CLA ) is supposed to spare glucose due to the milk fat‐depressing effect of the trans ‐10, cis ‐12 CLA isomer, and allows repartitioning nutrients despite an energy deficiency in early lactation. However, there is still a lack of knowledge in terms of the dynamic pattern of the glucose turnover in transition dairy cows. We hypothesized that dairy cows supplemented with CLA have an altered rate of glucose turnover and insulin sensitivity during early lactation. We conducted three consecutive hyperglycaemic clamps (HGC ) in weeks ?2, +2 and +4 relative to parturition in Holstein cows supplemented daily either with 70 g of lipid‐encapsulated CLA (6.8 g trans ‐10, cis ‐12 and 6.6 g of the cis ‐9, trans ‐11 CLA isomer; CLA ; n  = 11) or with 56 g of control fat ( CON ; n  = 11). From week ?3 up to week +4 relative to parturition, milk yield and dry matter intake (DMI ) were recorded daily, while body weight (BW ) and milk composition were obtained once weekly. Blood samples were taken once weekly and every 30 min during the HGC . Plasma was analysed for concentrations of glucose, fatty acids (FFA ), beta‐hydroxybutyrate (BHB ), insulin, triglycerides and cholesterol. The CLA supplementation did not affect performance and metabolic parameters except for BHB and cholesterol. Furthermore, insulin concentrations and insulin sensitivity were affected by treatment. During the HGC in early lactation, insulin response was lower and decrease in FFA and BHB greater compared with the HGC in week ?2 although glucose target concentration achieved during the steady‐state period was similar for all three HGC . Our findings in terms of insulin and cholesterol suggest that body reserves are preserved through CLA feeding without restraining animal's performance. Furthermore, CLA effects on cholesterol and triglyceride concentrations indicated beneficial effects on hepatic lipid export contributing to an improved efficiency of prevailing metabolites in circulation.     

The influence of rapid growth on sodium salicylate pharmacokinetics in male turkeys

The aim of this study was to assess the influence of growth on the pharmacokinetics of sodium salicylate (SS) in male turkeys. SS was administered intravenously at a dose of 50 mg/kg. Plasma drug concentrations were assessed by high‐performance liquid chromatography, and pharmacokinetic parameters were calculated by noncompartmental analysis. As the age increased from 6 to 13 weeks (body weight increase from 2.35 to 9.43 kg), median body clearance decreased from 1.34 to 0.87 ml/min/kg. This caused a significant increase in the median mean residence time from 3.42 to 4.44 hr. Elimination phase proved to be biphasic and two elimination half‐lives (T_1/2el) were distinguished. Whereas T_1/2el1 was found to increase with age by 128%, T_1/2el2 represented a later but faster and less age‐dependent phase of elimination (increase by 56% in the respective groups). Volume of distribution decreased with age. These effects may lead to different therapeutic response to SS in turkeys of different age and body weights.     

我国陆生动物疫病检测诊断技术标准化发展分析

动物疫病检测和诊断类标准是检测和判断动物疫病的主要依据。本文梳理了我国动物疫病检测和诊断技术标准的建设现状,并将其与我国《一、二、三类动物疫病病种名录》以及OIE《陆生动物疫病诊断和疫苗手册》和《中华人民共和国进境动物检疫疫病名录》中的陆生动物疫病目录进行对比。分析发现:我国动物疫病检测和诊断技术标准体系对我国一、二、三类动物疫病的覆盖率为92.6%,其中80%的标准有PCR和(或)ELISA检测方法 ;对OIE陆生动物疫病诊断和疫苗手册名录疫病的覆盖率为88.3%,其中73%的标准有PCR和(或)ELISA检测方法 ;对我国进境动物检疫名录疫病的覆盖率为87.7%,其中73%的标准有PCR和(或)ELISA检测方法。由此提出了提高我国动物疫病检测和诊断技术标准覆盖率,强化快速诊断检测技术标准化建设的建议。     

一起未经执业兽医注册从事诊疗活动案引发的思考

2017年成都市办理的一起未经兽医执业注册而进行犬只声带切除的行政处罚案件,引发了笔者对动物诊疗机构监督执法中"动物诊疗"范畴界定、动物诊疗许可证申请对象探究、"一事不再罚"原则的适用、"责令停业诊疗活动"与"责令停产停业"区别等问题的思考。本文以法学理论和相关法律规定为基础,对成都市动物诊疗机构执法实践的疑点与难点进行分析,提出执法人员应更准确适用法律法规,并呼吁尽快建立全国执业兽医查询系统的建议。     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Effect of banding or burdizzo castration on plasma testosterone, acute-phase proteins, scrotal circumference, growth, and health of bulls

The objective was to assess the effect of banding or burdizzo castration performed on farms on plasma testosterone, acute-phase proteins, scrotal circumferences, growth, and well-being of bulls. 243 Continental bulls (12 months; 399.2 ± 5.72 kg) from three different farms were allocated at random, after stratification on weight within breed type, to one of three treatment groups: banding castration (BAND; n = 80), burdizzo castration (BURD; n = 83), or controls (CON; n = 80). The castration methods were conducted under local anaesthesia, and tetanus toxoid vaccine and antibiotic were also injected at castration. BAND and BURD castrates had lower (P < 0.001) plasma testosterone concentration than control bulls, with no difference between BAND and BURD castrates on 28 d post-castration. From days 0 to 14 post-castration, BAND (P = 0.0002) and BURD (P < 0.0001) castrates had lower average daily gain (ADG) than CON bulls, no difference (P = 0.46) was found between BAND and BURD castrates. From days 15 to 28, BAND castrates had lower ADG compared with BURD castrates (P = 0.03) and CON bulls (P = 0.01), while no difference (P = 0.76) was found between BURD and CON. From days 29 to 56, BAND (P = 0.01) and BURD (P = 0.002) castrates had lower ADG than CON bulls, no difference (P = 0.55) was found between BAND and BURD. From days 57 to 84, the ADG of BAND castrates was not different compared with BURD castrates (P = 0.12) and CON bulls (P = 0.38), while BURD had lower (P = 0.02) ADG compared with CON. The integrated ADG from day 0 to 112 of BAND (P = 0.0001) and BURD (P = 0.02) groups were lower compared with CON, while there was no difference (P = 0.09) between BAND and BURD castrates. On d 14 post-castration, BAND castrates had lower scrotal temperature than BURD (P < 0.0001) and CON (P < 0.0001), and BURD castrates had greater (P < 0.006) scrotal temperature than CON; BAND castrates had lower scrotal latitudinal and longitudinal circumferences than BURD castrates (P < 0.001) and CON bulls (P < 0.001), and BURD castrates had greater (P < 0.001) scrotal latitudinal and longitudinal circumferences than CON bulls. BAND (P < 0.0001) and BURD (P = 0.01) castrates had greater glucose concentration than CON bulls, and BAND castrates had greater (P = 0.04) glucose concentration than BURD. In conclusion, BAND or BURD castration significantly reduced plasma testosterone concentration; reduced average daily weight gain mainly during the first 2 weeks, which was not compensated during the subsequent 16 weeks; increased withdrawal of stored energy and increased plasma protein concentration. BURD showed an advantage over BAND in growth during days 15 to 28 following castration.     

A Simple and Rapid PCR‐Based Method for Ostrich Sexing Using Micro Amounts of DNA

The aim of the present study is to identify ostrich sex by using polymerase chain reaction (PCR) on micro amounts of DNA from blood, bloodstain and feathers. Sixteen male and 18 female ostriches were used as test individuals. Genomic DNA as a template was extracted by the Chelex method. Ostrsex‐P1 and P2 primers were designed to perform PCR amplification on the template. PCR products were checked using agarose gel electrophoresis with ethidium bromide staining and ostrich sex was determined directly by the bands shown on the gel. The results demonstrate that ostrich sex can be determined by the extraction of DNA from as little as 0.0125 μl blood using Chelex, whereby the use of large amounts of organic solvents such as phenol and chloroform are unnecessary. In addition, it is possible to identify ostrich sex using micro amounts of DNA extracted from bloodstains and/or feathers. The use of feathers particularly avoids unwanted sampling problems such as the difficulty of collecting ostrich blood, the stress to the ostrich caused by bleeding, and the demand for a lot of manpower for ostrich restraint.