仔猪小肠肌间神经丛NDP阳性神经元形态的定量研究

采用小肠铺片NADPH黄递酶组化染色和细胞影像分析的方法对0、5、28日龄的仔猪小肠肌间神经丛神经元群体的形态参数进行定量测定。结果表明:随日龄增长,小肠肌间神经元胞体面积增大,出生后的头几天尤为明显。NDP阳性肌间神经元胞核面积平均值与胞体面积平均值呈正相关。胞体面积平均值在0日龄为234.98±23.48μm2,5日龄为346.30±33.07μm2,28日龄为364.17μm2;胞核面积平均值在0日龄为69.85±6.27μm2,5日龄为88.25±2.39μm2,28日龄为84.15μm2。核质比随日龄呈下降趋势,核质比在0日龄为0.298±0.003,5日龄为0.257±0.027,28日龄为0.231。上述测量值不仅表现出日龄差异,而且在同一日龄小肠的前后段亦有所不同。NDP阳性神经元胞体和胞核大小都以十二指肠最大,空肠前段次之,回肠最小。0日龄仔猪神经元胞体面积分布在100～300μm2之间的占70.79%,5日龄和28日龄胞体面积分布在100～400μm2之间的分别占64.81%和63.22%。肌间神经丛NDP阳性神经元主要为Dogiel 型神经元。     

A mutation confers Monochoria vaginalis resistance to sulfonylureas that target acetolactate synthase

Acetolactate synthase (ALS) is the target enzyme for four distinct families of compounds: sulfonylureas (SUs), imidazolinones, triazolopyrimidine sulfonanilides, and pyrimidinyl oxybenzoates. We cloned and sequenced the fragments encoding ALS genes from biotypes of Monochoria vaginalis susceptible (S) and resistant (R) to SU-herbicides. The nucleotide sequences of the 39 bp Domain A region for R M. vaginalis biotype differed from that of the S biotype by a single nucleotide substitution at variable Pro codon of Domain A (CCT to TCT), predicting a Pro in the S but a Ser in the R biotype. No nucleotide differences between S and R M. vaginalis were observed in Domain D. We suggest that the amino acid substitution at Domain A region is responsible for resistance to SU-herbicides in M. vaginalis collected from Ushiku City, Ibaraki Prefecture, Japan.     

小叶锦鸡儿根瘤固氮活性的动态变化

对多年生小叶锦鸡儿不同生育期根瘤固氮能力、不同树龄小叶锦鸡儿根瘤固氮能力进行调查,结果表明:开花期固氮酶活性最高,此时土壤含氮量和植株含氮量最高;新瘤多发生于果后营养期,植株生物量的峰值出现在结实期;二年生和五年生植株的根瘤固氮能力强于三年、四年生植株,随生长年限的延长,植株生物量逐渐增大,含氮量呈小幅度下降趋势。     

Canine GM2‐Gangliosidosis Sandhoff Disease Associated with a 3‐Base Pair Deletion in the HEXB Gene

Background

GM2‐gangliosidosis is a fatal neurodegenerative lysosomal storage disease (LSD) caused by deficiency of either β‐hexosaminidase A (Hex‐A) and β‐hexosaminidase B (Hex‐B) together, or the GM2 activator protein. Clinical signs can be variable and are not pathognomonic for the specific, causal deficiency.

Objectives

To characterize the phenotype and genotype of GM2‐gangliosidosis disease in an affected dog.

Animals

One affected Shiba Inu and a clinically healthy dog.

Methods

Clinical and neurologic evaluation, brain magnetic resonance imaging (MRI), assays of lysosomal enzyme activities, and sequencing of all coding regions of HEXA, HEXB, and GM2A genes.

Results

A 14‐month‐old, female Shiba Inu presented with clinical signs resembling GM2‐gangliosidosis in humans and GM1‐gangliosidosis in the Shiba Inu. Magnetic resonance imaging (MRI) of the dog's brain indicated neurodegenerative disease, and evaluation of cerebrospinal fluid (CSF) identified storage granules in leukocytes. Lysosomal enzyme assays of plasma and leukocytes showed deficiencies of Hex‐A and Hex‐B activities in both tissues. Genetic analysis identified a homozygous, 3‐base pair deletion in the HEXB gene (c.618‐620delCCT).

Conclusions and Clinical Importance

Clinical, biochemical, and molecular features are characterized in a Shiba Inu with GM2‐gangliosidosis. The deletion of 3 adjacent base pairs in HEXB predicts the loss of a leucine residue at amino acid position 207 (p.Leu207del) supporting the hypothesis that GM2‐gangliosidosis seen in this dog is the Sandhoff type. Because GM1‐gangliosidosis also exists in this breed with almost identical clinical signs, genetic testing for both GM1‐ and GM2‐gangliosidosis should be considered to make a definitive diagnosis.     

Bioavailability of suppository acetaminophen in healthy and hospitalized ill dogs

To determine the plasma pharmacokinetics of suppository acetaminophen (APAP) in healthy dogs and clinically ill dogs. This prospective study used six healthy client‐owned and 20 clinically ill hospitalized dogs. The healthy dogs were randomized by coin flip to receive APAP orally or as a suppository in crossover study design. Blood samples were collected up to 10 hr after APAP dosing. The hospitalized dogs were administered APAP as a suppository, and blood collected at 2 and 6 hr after dosing. Plasma samples were analyzed by ultra‐performance liquid chromatography with triple quadrupole mass spectrometry. In healthy dogs, oral APAP maximal concentration (C_MAX=2.69 μg/ml) was reached quickly (T_MAX=1.04 hr) and eliminated rapidly (T1/2 = 1.81 hr). Suppository APAP was rapidly, but variably absorbed (C_MAX=0.52 μg/ml T_MAX=0.67 hr) and eliminated (T_1/2 = 3.21 hr). The relative (to oral) fraction of the suppository dose absorbed was 30% (range <1%–67%). In hospitalized ill dogs, the suppository APAP mean plasma concentration at 2 hr and 6 hr was 1.317 μg/ml and 0.283 μg/ml. Nonlinear mixed‐effects modeling did not identify significant covariates affecting variability and was similar to noncompartmental results. Results supported that oral and suppository acetaminophen in healthy and clinical dogs did not reach or sustain concentrations associated with efficacy. Further studies performed on different doses are needed.     

The construction and application of a population physiologically based pharmacokinetic model for methadone in Beagles and Greyhounds

Methadone is an opioid analgesic in veterinary and human medicine. To help develop appropriate pain management practices and to develop a quantitative model for predicting methadone dosimetry, a flow‐limited multiroute physiologically based pharmacokinetic (PBPK) model for methadone in dogs constructed with Berkeley Madonna? was developed. The model accounts for intravenous (IV), subcutaneous (SC), and oral administrations, and compartmentalizes the body into different components. This model was calibrated from plasma pharmacokinetic data after IV administration of methadone in Beagles and Greyhounds. The calibrated model was evaluated with independent data in both breeds of dogs. One advantage of this model is that most physiological parameter values for Greyhounds were taken directly from the original literature. The developed model simulates available pharmacokinetic data for plasma concentrations well for both breeds. After conducting regression analysis, all simulated datasets produced an R ² > 0.80 when compared to the measured plasma concentrations. Comparative analysis of the dosimetry of methadone between the breeds suggested that Greyhounds had ~50% lower 24‐hr area under the curve (AUC) of plasma or brain concentrations than in Beagles. Furthermore, population analysis was conducted with this study. This model can be used to predict methadone concentrations in multiple dog breeds using breed‐specific parameters.     

Effect of gut stress induced by oxidized wheat gluten on the growth performance,gut morphology and oxidative states of broilers

This study was to investigate the effect of oxidized wheat gluten (OG) on growth performance, gut morphology and its oxidative states of broilers. One hundred and eighty‐day‐old male broilers (10 chicks/pen) were randomly allocated into three dietary treatments: control diet (CON), diet with 8% wheat gluten (WG) and diet with 8% OG with six pens/treatment. Body weight (BW) (21 and 35 days) and average daily gain (ADG) (1–21 days and 22–35 days) decreased (p < .05) and feed conversion ratio (FCR) (1–21 days and 22–35 days) increased (p < .05) in OG treatment. Feed intake (FI) decreased (p < .05) in WG and OG treatments during 22–35 days. However, FI was not influenced by dietary treatments during 1–21 days (p > .05). The OG‐fed broilers had a lower faecal pH value (p < .05) and higher faecal moisture content (p < 05) at 14, 21, 28 and 35 days. Villus height, crypt depth and V/C value were not different (p > .05) among treatments at 21 and 35 days. Lipid peroxidation (LPO) (21 and 35 days) and malondialdehyde (MDA) (35 days) content in crop of OG treatment increased (p < .05). Oxidized glutathione (GSSG) (21 days), LPO (21 and 35 days) and MDA (21 and 35 days) content in ileum of OG treatment increased (p < .05). The reduced glutathione/oxidized glutathione (GSH/GSSG) (21 days) and (GSH) (35 days) in ileum of OG treatment decreased (p < .05). The present findings indicate that OG might be a stressor for broiler gut, which could induce oxidative stress both in crop and in ileum, and the diarrhoea as well. The growth performance of broiler was consequently depressed.     

Methyl donors dietary supplementation to gestating sows diet improves the growth rate of offspring and is associating with changes in expression and DNA methylation of insulin‐like growth factor‐1 gene

The study aimed to investigate the effects of maternal dietary methyl donors on the performance of sows and their offspring, and the associated hepatic insulin‐like growth factor‐1 (IGF‐1) expression of the offspring. A total of 24 multiparous sows were randomly fed the control (CON) or the CON diet supplemented with methyl donors (MD) at 3 g/kg betaine, 15 mg/kg folic acid, 400 mg/kg choline and 150 μg/kg VB₁₂, from mating until delivery. After farrowing, sows were fed a common lactation diet through a 28‐days lactation period and six litters per treatment were selected to be fed until at approximately 110 kg BW. Maternal MD supplementation resulted in greater birthweight (p < 0.05) and increased the piglet weights (p < 0.01) and litter weights (p < 0.05) at the age of day 28, compared with that in CON group. The offspring pigs in the MD group had greater ADG (p < 0.05) and tended to lower F:G ratio (p = 0.07) compared with that of CON group from day 28 to 180 of age. The offspring pigs from MD group had greater serum IGF‐1 concentrations and expressions of hepatic IGF‐1 gene and muscular IGF‐1 receptor (IGF‐1r) protein at birth (p < 0.05), and greater hepatic IGF‐1 protein (p = 0.03) and muscular IGF‐1r gene expressions (p < 0.05) at slaughter, than that from the CON group. Moreover, the methylation at the promoter of IGF‐1 gene in the liver of newborn piglets and finishing pigs was greater in the MD group than that of the CON group (p < 0.05). In conclusion, maternal MD supplementation throughout gestation could enhance the birthweight and postnatal growth rate of offspring, associated with an increased expression of the IGF‐1 gene and IGF‐1r, as well as the altered DNA methylation of IGF‐1 gene promotor.     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

基于"互联网+"的草业企业经营管理模式——以草都易牧连锁超市为例

在"互联网+"大潮改造着各行各业的背景下,新时期的草业企业经营管理模式正发生着深刻的变化。基于此,以草业电商的实例——草都易牧连锁超市为例,分析其经营管理模式,并运用成本效益分析法从理论上探讨其经济可行性。结果表明,草都易牧连锁超市的经营管理模式为"互联网+草业"模式。该模式能将牧区饲草料的需求和供给对接起来,增加参与其中的线上牧户与入驻企业的利润,实现二者的共赢,在理论上具有经济可行性。草都易牧连锁超市的大胆尝试是草业企业在当前经济社会环境下一种重要的"互联网+草业"创新模式。通过对"互联网+"在草业中具体应用的深入探讨,认为草业企业的"互联网+草业"模式多主体分析、实证分析以及草产业链尺度分析或将成为"互联网+草业"未来的3个重点研究方向。