一株鸭源禽流感病毒（H9N2）全基因序列分析及其排毒规律的研究

为研究一株鸭源H9N2亚型禽流感病毒(JN1株)全基因组的分子特征和遗传进化情况以及感染雏鸭后的排毒规律,对其生物学特性、全基因组序列以及遗传进化进行了分析,通过点眼滴鼻方式感染3周龄健康雏鸭,利用建立的实时荧光定量PCR(RT-PCR)检测鸭的排毒规律,并测定血清抗体效价。JNI株的鸡胚半数感染量为10~(-7.54)/0.2mL,鸡胚最小致死量的平均死亡时间为72h,静脉致病指数为0.266,抗原相关系数为0.47。系统发育分析表明,JN1株的HA与CK/HK/G9/97和DK/HK/Y280/97在同一分支上,NA、M、NP、PA、PB1、PB2基因与SH/F/98同源性较高,NS基因则与H5N1亚型禽流感病毒进化关系较近。HA的裂解位点为PSRSSR↓GL,存在6个潜在糖基化位点,在234位的氨基酸残基为L,NA基因颈部存在缺失。雏鸭在感染该病毒后,饮食正常,精神良好,3~17d均有排毒,咽拭子和泄殖腔棉拭子排毒规律基本一致,分别在第3天和第13天出现排毒高峰。该H9N2亚型禽流感病毒属欧亚大陆分支,为低致病性禽流感,病毒可通过呼吸道和泄殖腔向外界排毒,排毒时间较长。     

IBV AH1-99分离株N基因的克隆及序列分析

利用RT-PCR技术成功扩增出IBV AH1-99分离株的N基因全长cNDA,将其克隆到pMD18-T载体上。经酶切分析、PCR鉴定及核苷酸序列测定,成功获得该分离株N基因的重组质粒。序列分析结果表明,分离株N基因全长1 230个核苷酸,编码409个氨基酸。同部分IBV参考毒株相比,核苷酸同源性为85.8%~89.9%,氨基酸同源性为86.6%~91.0%,在进化关系树中,AH1-99与国内分离株X、LX4亲缘关系较近。     

Effect of contrast media formulation on computed tomography angiographic contrast enhancement.

The characteristics of contrast media formulation (mgI/ml, osmolarity, and viscosity) are generally not considered important in computed tomography (CT) angiography in animals. The purpose of this study was to assess the contrast effect in CT angiography as a function of contrast media formulation, with a constant iodine dose. The contrast effects of three contrast media with different iodine concentrations were compared by administering identical iodine dosages (mgI/kg). The contrast effects of the three contrast media differed, and the area under the time-attenuation curve of iohexol 350 mgI/ml, which had the highest iodine concentration, was the lowest. It was hypothesized that the contrast effect of a contrast medium decreases with higher iodine concentration because of the high amount of residual iodine present in the circulatory system from the injection site to the portion immediately before the great vessels. In addition, the influence of osmotic dilution on contrast media with high osmolarity was also considered. In conclusion, the contrast effect varies with different contrast media formulations, even when the same iodine dosage is administered.     

Effects of a low dose infusion of racemic and S-ketamine on the nociceptive withdrawal reflex in standing ponies

ObjectiveTo investigate the effect of plasma concentrations obtained by a low dose constant rate infusion (CRI) of racemic ketamine or S-ketamine on the nociceptive withdrawal reflex (NWR) in standing ponies.Study designProspective, blinded, cross-over study.AnimalsSix healthy 5-year-old Shetland ponies.MethodsPonies received either 0.6 mg kg⁻¹ racemic ketamine (group RS) or 0.3 mg kg⁻¹ S-ketamine (group S) intravenously (IV), followed by a CRI of 20 μg kg⁻¹minute⁻¹ racemic ketamine (group RS) or 10 μg kg⁻¹minute⁻¹ S-ketamine (group S) for 59 minutes. The NWR was evoked by transcutaneous electrical stimulation of a peripheral nerve before drug administration, 15 and 45 minutes after the start of the bolus injection and 15 minutes after the end of the CRI. Electromyographic responses were recorded and analysed. Arterial blood was collected before stimulation and plasma concentrations of ketamine and norketamine were measured enantioselectively using capillary electrophoresis. Ponies were video recorded and monitored to assess drug effects on behaviour, heart rate (HR), mean arterial blood pressure (MAP) and respiratory rate.ResultsThe NWR was significantly depressed in group RS at plasma concentrations between 20 and 25 ng mL⁻¹ of each enantiomer. In group S, no significant NWR depression could be observed; plasma concentrations of S-ketamine (9–15 ng mL⁻¹) were lower, compared to S-ketamine concentrations in group RS, although this difference was not statistically significant. Minor changes in behaviour, HR and MAP only occurred within the first 5–10 minutes after bolus drug administration in both groups.ConclusionAntinociceptive activity in standing ponies, demonstrated as a depression of the NWR, could only be detected after treatment with racemic ketamine. S-ketamine may have lacked this effect as a result of lower plasma concentrations, a more rapid metabolism or a lower potency of S-ketamine in Equidae so further investigation is necessary.     

Naltrexone does not affect isoflurane minimum alveolar concentration in cats

ObjectiveTo test whether naltrexone, an opioid receptor antagonist, affects the minimum alveolar concentration (MAC) of isoflurane in cats, a species that is relatively resistant to the general anesthetic sparing effects of most opioids.Study designRandomized, crossover, placebo-controlled, blinded experimental design.AnimalsSix healthy adult cats weighing 4.9 ± 0.7 kg.MethodsThe cats were studied twice. In the first study, baseline isoflurane MAC was measured in duplicate. The drug (saline control or 0.6 mg kg^?1 naltrexone) was administered IV every 40–60 minutes, and isoflurane MAC was re-measured. In the second study, cats received the second drug treatment using identical methods 2 weeks later.ResultsIsoflurane MAC was 2.03 ± 0.12% and was unchanged from baseline following saline or naltrexone administration.Conclusion and clinical relevanceMinimum alveolar concentration was unaffected by naltrexone. Because MAC in cats is unaffected by at least some mu-opioid agonists and antagonists, spinal neurons that are directly modulated by mu-opioid receptors in this species cannot be the neuroanatomic sites responsible for immobility from inhaled anesthetics.     

Plasma endotoxin concentration in horses: a methods study

Background: Accurate determination of plasma endotoxin concentration is critical for ex vivo and in vitro cellular and molecular studies of endotoxemia in horses. However, reports are conflicting with respect to anticoagulant, handling, and sample preparation.

Objective:

Objective: The purpose of this study was to determine the effect of blood sample fraction and handling time on measurement of endotoxin concentration in horses.

Methods:

Methods: Whole blood, anticoagulated with 3.8% (0.12 M) sodium citrate (9:1), was collected from 5 healthy horses. Whole blood (WB), platelet-rich plasma (PRP), and platelet-poor plasma (PPP) were spiked with endotoxin (2 EU/mL). Endotoxin-spiked WB samples were centrifuged immediately to generate PRP for measurement. Endotoxin concentration was subsequently measured by Limulus amebocyte assay at 0, 15, 30, 45, and 60 minutes. Assays were performed in triplicate and results were analyzed using Student's t -test, with significance set at P < .05.

Results:

Results: Mean endotoxin concentrations in 2 EU/mL-spiked WB were significantly different from those in PPP at all time points tested. Recovery of endotoxin in PRP generated from WB was significantly diminished after just 15 minutes.

Conclusion:

Conclusion: PRP generated from WB is significantly more reliable than PPP in determining endotoxin concentration ex vivo. Measurement of endotoxin in PRP generated from WB was significantly diminished after 15 min, identifying a time frame within which to process blood samples for endotoxin analysis.     

H9亚型禽流感病毒钦州株HA基因的克隆与遗传进化

采集钦州活禽交易市场的鸡气管和泄殖腔的棉拭予样品,用H9亚型分型引物进行PCR初步筛选,阳性样品经SPF鸡胚尿囊腔接种分离病毒,通过RT-PCR方法扩增HA基因,并将其克隆到pMD—18T载体后进行序列测定和分析。结果表明,获得1株H9亚型禽流感病毒命名为：A／Chicken／Guangxi／qz40／2009（简称qz40）。测序结果表明qz40的HA基因片段全长1683bp,编码560个氨基酸;序列同源性比较结果表明,该毒株与参考毒株的核苷酸序列同源性为82．8％．99．9％,推导氨基酸同源性为87．5％．99．6％;HA基因的裂解位点氨基酸顺序为RSSRIGLF,为低致病性毒株;含有7个潜在的N-糖基化位点,其中5个位于HAl部分、2个位于HA2部分;基于H9亚型HA基因的进化树分析表明,qz40株属于欧亚种系的A／Chicken／Beijing／1／94（Ck／Bei—like）群系。     

The Effect of Levothyroxine Treatment on Resting Energy Expenditure of Hypothyroid Dogs

Thirty adult, client-owned dogs were diagnosed with hypothyroidism based on history, physical examination findings, hematologic and biochemical abnormalities, thyrotropin (TSH) response testing, endogenous canine thyrotropin (cTSH) concentration, or both, and total serum thryoxine concentration. All dogs received levothyroxine (L-thyroxine) at an initial dose of 22 μg/kg PO sid in either a tablet (13 dogs) or chewable form (17 dogs). Energy expenditure of each dog during apparent rest was estimated with an open-flow indirect calorimetry system by determining the rates of carbon dioxide production and oxygen consumption. Energy expenditure of apparent rest (EE) was lower in untreated hypothyroid dogs compared with reference values for EE. After treatment with L-thyroxine, EE of the hypothyroid dogs was significantly ( P < .05) higher than pretreatment values.     

水淹胁迫对狗牙根光合、生长及营养元素含量的影响

以狗牙根当年生扦插苗为试验材料,采用盆栽控制实验,设置对照组(CK)、表土水淹组(SF)和全淹组(TF),研究水淹对狗牙根光合、生长、生物量和营养元素含量的影响。结果显示,1)与CK相比,SF组的狗牙根净光合速率、气孔导度、蒸腾速率和气孔限制值均显著降低,胞间CO₂浓度显著增加,但水分利用效率与CK组差异不显著。2)SF组分枝数、叶片数、根生物量、叶生物量和根冠比均显著低于CK组,而株高、基径、茎生物量和总生物量与CK组差异不显著;TF组除根冠比与CK组无显著差异外,株高、基径、分枝数、叶片数、根生物量、茎生物量和总生物量均显著低于CK组。3)与CK组相比,SF和TF组根的N、P、K、Fe、Mn含量均呈上升趋势,Cu含量显著降低;叶片中,SF组P、K、Cu含量显著下降,Fe、Mn含量显著升高。研究表明,水淹显著影响了狗牙根光合、生长及营养元素含量。狗牙根通过适度降低叶片P、K、Cu含量及升高Fe、Mn含量促进狗牙根叶片净光合速率的降低。然而狗牙根可通过提高对N、P、K等营养元素的吸收,促进其生长,维持较高的净光合速率,以有效应对水淹环境;其次,全淹环境中,狗牙根叶片衰老、凋落,叶片中N、P等营养元素可直接进入水体,存在增加水体富营养化的潜在风险,因此,应加强对三峡库区消落带狗牙根植被的刈割收获管理,以尽可能减少由此引起的二次污染。