Chitosan nanoparticles enhance developmental competence of in vitro-matured porcine oocytes

Oxidative stress is inevitable as it is derived from the handling, culturing, inherent metabolic activities and medium supplementation of embryos. This study was performed to investigate the protective effect of chitosan nanoparticles (CNPs) on oxidative damage in porcine oocytes. For this purpose, cumulus–oocyte complexes (COCs) derived from porcine slaughterhouse ovaries were exposed to different concentrations of CNPs (0, 10, 25 and 50 µg/ml) during in vitro maturation (IVM). Oocytes treated with 25 µg/ml CNPs showed significantly higher levels of GSH, along with a significant reduction in ROS levels compared to control, CNPs10 and CNPs50 groups. In parthenogenetic embryo production, the maturation rate was significantly higher in the CNPs25 group than that in the control and all other treated groups. In addition, when compared to the CNPs50 and control groups, CNPs25-treated oocytes showed significantly higher cleavage and blastocyst development rates. The highest concentration of CNPs reduced the total cell number and ratio of ICM: TE cells in parthenogenetic embryos, suggesting that there is a threshold where benefits are lost if exceeded. In cloned embryos, the CNPs25 group, as compared to all other treated groups, showed significantly higher maturation and cleavage rates. Furthermore, the blastocyst development rate in the CNPs25-treated group was significantly higher than that in the CNPs50-treated group, as was the total cell number. Moreover, we found that cloned embryos derived from the CNPs25-treated group showed significantly higher expression levels of Pou5f1, Dppa2, and Ndp52il genes, compared with those of the control and other treated groups. Our results demonstrated that 25 µg/ml CNPs treatment during IVM improves the developmental competence of porcine oocytes by reducing oxidative stress.     

子午岭黑山羊与辽宁绒山羊产肉性能、肉品质、肌肉营养成分和脂肪酸含量比较

1990年,甘肃省庆阳市开始引入辽宁绒山羊对子午岭黑山羊进行杂交改良,但目前尚不清楚2个品种在脂肪酸含量、肌肉营养成分等方面的差异,影响了杂交改良效果。试验旨在分析两个绒山羊品种的产肉性能、肉品质、肌肉营养成分和脂肪酸含量差异,为绒山羊的杂交改良提供理论依据。本研究选取相同饲养管理条件下、9月龄的子午岭黑山羊和辽宁绒山羊公羊各5只,测定其屠宰性能以及背最长肌、前腿肌和后腿肌处的肉品质、脂肪酸含量和肌肉营养成分。结果表明：子午岭黑山羊的胴体重、屠宰率、净肉重、净肉率、眼肌面积、GR值、剪切力和滴水损失低于辽宁绒山羊（P<0.05）,但其肌肉的平均亮度值、色度值、pH₁和pH₂₄高于辽宁绒山羊（P<0.01）。营养成分测定结果表明,子午岭黑山羊肌肉的水分和粗灰分含量高于辽宁绒山羊（P<0.05）,但肌内脂肪和粗蛋白含量低于辽宁绒山羊。在2个山羊品种的肌肉中均检测到11 种饱和脂肪酸（SFA,以棕榈酸和硬脂酸为主）、10种多不饱和脂肪酸（PUFA,以亚油酸和顺-11,14-二十碳二烯酸为主）和6种单不饱和脂肪酸（MUFA,以油酸为主）,子午岭黑山羊肉中的SFA、PUFA、n-3 PUFA、n-6 PUFA含量和PUFA/SFA值均高于辽宁绒山羊（P<0.01）,但MUFA含量低于辽宁绒山羊（P<0.01）。结果表明,辽宁绒山羊有更高的产肉力,但子午岭黑山羊肌肉品质和营养成分更佳,脂肪酸组成和含量更符合人类健康膳食标准。     

基于转录组测序分析研究枸芪多糖对育肥猪肠黏膜免疫功能的作用机理

本试验旨在研究枸芪多糖对育肥猪肠黏膜免疫调控的关键基因,阐明其对肠黏膜免疫功能的可能作用机理。选用80~90日龄的健康育肥母猪180头,随机分为2组,每组6个重复,每个重复15头猪。对照组饲喂基础饲粮,试验组在基础饲粮中添加0.1%的枸芪多糖,试验期90 d。采集育肥猪的空肠肠段,提取RNA后进行转录组测序分析差异免疫基因的表达。结果显示:1)与对照组相比,饲粮中添加枸芪多糖差异表达的免疫基因有75个,其中有26个上调基因和49个下调基因。2)基因本体(GO)富集分析显示差异表达基因主要参与免疫调节、免疫应答和应激反应,信号通路分析显示差异表达基因主要富集在以白细胞介素-17(IL-17)信号通路、NOD样受体信号通路为主的免疫信号通路。3)通过对肠黏膜差异表达的关键基因原癌基因Jun(JUN)和原癌基因Fos(FOS)的探究可见,与对照组相比,试验组白细胞介素-18(IL-18)、补体5(C5)、JUN和FOS的基因表达受到抑制,Jun和Fos的蛋白表达显著降低(P<0.05)。综上所述,在饲粮中添加枸芪多糖可提高肥育猪的肠黏膜免疫功能。枸芪多糖通过调节JUN、FOS、IL-18等免疫相关基因的表达,影响IL-17和NOD样受体信号通路,以此来抑制炎症反应。     

Myocarditis caused by naturally acquired canine distemper virus infection in 4 dogs

Canine distemper virus (CDV) has long been recognized as a cause of myocarditis; however, cases of myocarditis caused by naturally acquired CDV infection have been reported only rarely in dogs. We describe here our retrospective study of naturally acquired systemic CDV infection in 4 dogs, 4–7 wk old, that had myocarditis, with myocardial necrosis and fibrosis. One of the 4 dogs had intracytoplasmic eosinophilic inclusion bodies in cardiomyocytes. Other lesions included bronchointerstitial pneumonia (4 of 4), necrotizing hepatitis (2 of 4), splenic lymphoid necrosis (2 of 4), encephalitis (1 of 3; brain was not submitted in 1 case), and necrotizing gastroenteritis (1 of 4). The presence of CDV in the heart was confirmed by immunohistochemistry in all 4 dogs.     

Cerebrospinal fluid flow in normal beagle dogs analyzed using magnetic resonance imaging

BackgroundDiseases related to cerebrospinal fluid flow, such as hydrocephalus, syringomyelia, and Chiari malformation, are often found in small dogs. Although studies in human medicine have revealed a correlation with cerebrospinal fluid flow in these diseases by magnetic resonance imaging, there is little information and no standard data for normal dogs.ObjectivesThe purpose of this study was to obtain cerebrospinal fluid flow velocity data from the cerebral aqueduct and subarachnoid space at the foramen magnum in healthy beagle dogs.MethodsSix healthy beagle dogs were used in this experimental study. The dogs underwent phase-contrast and time-spatial labeling inversion pulse magnetic resonance imaging. Flow rate variations in the cerebrospinal fluid were observed using sagittal time-spatial labeling inversion pulse images. The pattern and velocity of cerebrospinal fluid flow were assessed using phase-contrast magnetic resonance imaging within the subarachnoid space at the foramen magnum level and the cerebral aqueduct.ResultsIn the ventral aspect of the subarachnoid space and cerebral aqueduct, the cerebrospinal fluid was characterized by a bidirectional flow throughout the cardiac cycle. The mean ± SD peak velocities through the ventral and dorsal aspects of the subarachnoid space and the cerebral aqueduct were 1.39 ± 0.13, 0.32 ± 0.12, and 0.76 ± 0.43 cm/s, respectively.ConclusionsNoninvasive visualization of cerebrospinal fluid flow movement with magnetic resonance imaging was feasible, and a reference dataset of cerebrospinal fluid flow peak velocities was obtained through the cervical subarachnoid space and cerebral aqueduct in healthy dogs.     

Rapidly quantitative detection of Nosema ceranae in honeybees using ultra-rapid real-time quantitative PCR

BackgroundThe microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen.ObjectivesThe present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees.MethodsA procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration.ResultsUR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min.ConclusionsUR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.     

低血糖生成指数液态奶的开发及测试

为获得营养均衡的调制牛奶,满足人体的多种营养需求,添加多种维生素和矿物质,设计出一款适用于糖尿病人、亚健康人群、体重管理、皮肤管理等人群饮用的低血糖生成指数（Glycemic Index,GI）的牛奶。稳定性试验结果表明,静置12 天后,牛奶无显著脂肪上浮,体系稳定性良好;人体GI值试验结果表明,调制牛奶的餐后血糖值上升和下降均较葡萄糖餐后血糖值平稳,有较好稳定餐后血糖的效果,表明开发的调制牛奶为低GI食品（GI值=22.86）。人体饱腹感试验结果显示,和普通纯牛奶对比,调制牛奶具有显著提高饱腹感的效果。     

新疆规模化奶牛场牛群结构调查与问题分析

为加快奶牛养殖由数量型向质量效益型转变,促进奶业提质增效高质量发展,助力新疆“十四五”规划开好局、起好步,笔者所在团队对新疆8 个地州、35 个县、85 家规模化奶牛场进行了实地查看走访、数据采集,重点对牛群结构及生产技术参数进行分析。结果发现,大部分奶牛场的牛群结构都存在一些较为突出的共性问题,影响了奶牛场的经营效益与长期发展。本文对问题进行了汇总,并提出了有效的改进措施,旨在引起奶牛场经营者的重视,同时也为日后有针对性地开展技术服务提供参考。     

The effects of maxillary nerve block,ethmoidal nerve block and their combination on cardiopulmonary responses to nasal stimulation in anesthetized Beagle dogs

ObjectiveTo describe an approach for ethmoidal nerve block (E_BLOCK) and to compare the effects of a maxillary nerve block (M_BLOCK), E_BLOCK and their combination (M-E_BLOCK) on heart rate (HR), systolic (SAP), mean (MAP), diastolic (DAP) arterial pressures and respiratory rate (f_R) during nasal stimulation in dogs.Study designProspective, blinded, randomized, crossover placebo-controlled study.AnimalsBeagle dogs (five cadavers, nine live dogs), with a median (interquartile range) weight of 10.5 (10.3–11.0) kg.MethodsThe accuracy of iohexol injections (each 1 mL) at the maxillary and ethmoidal foramina in cadavers was evaluated using computed tomography. Then, anesthetized dogs were administered four bilateral treatments separated by 1 week, saline or 2% lidocaine 1 mL per injection: injections of saline at the maxillary and ethmoidal foramina (Control), injections of lidocaine at the maxillary foramina and saline at the ethmoidal foramina (M_BLOCK), injections of saline at the maxillary foramina and lidocaine at the ethmoidal foramina (E_BLOCK) and injections of lidocaine at all foramina (M-E_BLOCK). The ventral nasal meatus was bilaterally stimulated using cotton swabs, and HR, SAP, MAP, DAP and f_R were continuously recorded. Values for each variable were compared before and after stimulation using Wilcoxon signed-rank test. Changes in variables among treatments were analyzed using Mann–Whitney U and Kruskal–Wallis tests (p ≤ 0.05).ResultsComputed tomography revealed iohexol distribution around the openings of the target foramina in all cadavers. In living dogs, HR, SAP, MAP, DAP and f_R significantly increased after stimulation within each treatment (p < 0.03). Physiologic responses were significantly attenuated, but not absent, in the M-E_BLOCK [HR (p = 0.019), SAP, MAP, DAP and f_R (all p ≤ 0.001)] compared with those in the Control.Conclusions and clinical relevanceConcurrent injections of lidocaine at the maxillary and ethmoidal foramina attenuated HR, arterial pressure and f_R responses to nasal stimulation in Beagle dogs.     

松辽黑猪METTL23基因多态性及其与繁殖性状的关联分析

为探究松辽黑猪METTL23基因多态性及其与繁殖性状的关联性,试验选取178头松辽黑猪母猪为研究对象,利用Sanger直接测序法查找METTL23基因外显子1~5的单核苷酸多态性(SNP),使用SPSS 19.0软件分析METTL23基因SNP位点与松辽黑猪繁殖性状(总产仔数、产活仔数、断奶仔猪数、初生重、3周龄重、断奶重、乳头数)的关联性。结果显示,仅在松辽黑猪METTL23基因外显子4发现1个SNP位点,命名为A62G,在A62G位点上检测到3种基因型,分别是AA、AG和GG。GG和G分别为优势基因型和优势等位基因,且A62G位点处于Hardy-Weinberg平衡状态(P>0.05)。该位点的遗传纯合度(Ho)为0.6516,高于遗传杂合度(He,0.3484),说明其在松辽黑猪群体中的变异较小;多态信息含量(PIC)为0.2877,属于中度多态位点(0.25<PIC<0.5),说明该遗传标记能够提供一定量的遗传信息。关联分析结果表明,GG基因型个体总产仔数、产活仔数和断奶仔猪数均显著或极显著高于AA基因型个体(P<0.05;P<0.01),总产仔数还显著高于AG基因型个体(P<0.05);AG基因型个体断奶仔猪数显著高于AA基因型个体(P<0.05);初生重、3周龄重、断奶重、乳头数各基因型间差异均不显著(P>0.05)。综上所述,METTL23基因外显子4存在多态性位点A62G,该位点与松辽黑猪产仔数存在显著关联,可作为松辽黑猪产仔数的遗传标记,用于分子育种。