Baclofen intoxication in a dog successfully treated with hemodialysis and hemoperfusion coupled with intensive supportive care

Objective: To describe a case of confirmed baclofen intoxication in a dog that was successfully treated with hemodialysis and hemoperfusion (HD/HP) and to report the serum baclofen kinetics. Case summary: A 2.5‐year‐old, 23 kg, spayed female Brittany Spaniel‐mix was treated after ingesting 21‐52 mg/kg of baclofen. The dog was comatose and was receiving manual ventilation at the time of presentation. Extracorporeal HD/HP was started 10 hours after admission. Within 3 hours of starting HD/HP the dog began initiating breaths and was extubated 18 hours after admission. Serial serum samples that were obtained during the first 24 hours of hospitalization were later analyzed for baclofen concentrations. The dog had elevated creatine phosphokinase and liver enzymes that correlated with an agitated recovery period. The dog had thrombocytopenia that resolved by 10 days after presentation. New or unique information provided: HD/HP shortened the baclofen serum elimination half‐life from 5 to 1.5 hours in the initial 2 hours of treatment. The intrinsic elimination rate constant (K_intr) for this dog was 0.138/hour and the total elimination rate constant (K_tot) during the first 2 hours of HD/HP treatment was 0.458/hour. In this dog, HD/HP was an effective method for rapidly decreasing serum baclofen concentration after an acute overdose.     

牛羊猪肉组织中伊维菌素残留量检测HPLC法的改进

对牛、羊、猪肉组织中伊维菌素残留的检测方法进行了改进，样品用乙腈提取，用正己烷脱脂并于50℃减压蒸干。残留物用乙酸乙酯溶解，过C18SPE柱，用乙酸乙酯-甲醇（1：1）溶液洗脱；收集流出液和洗脱液，合并蒸干后于（50±3）℃减压干燥20min；残留物用N-甲基咪唑溶解并与三氟乙酸酐-乙腈（1：2）溶液于（67±2）℃衍生1h。用配荧光检测器的高效液相色谱仪检测。本方法平均回收率为（102．0±6．6）％，批内RSD〈10％，伊维菌素浓度在5～400ng／mL时，呈线性关系，r=0．9997。检测限为2μg／kg。     

金霉素和代谢物差向金霉素在肉鸡肌肉和肝脏中的残留分布

为了研究金霉素(CTC)、差向金霉素(ECTC)在肉鸡肌肉、肝脏中的残留分布情况,样品经提取液提取后过Oasis HLB SPE小柱净化,采用电喷雾正离子模式(ESI+)和多反应监测模式(MRM),用高效液相色谱-串联质谱仪检测;试验组鸡按体重以50 mg/(kg·d)剂量内服CTC,连续5 d,每天给药1次。结果表明:该方法测定肉鸡肌肉和肝脏中CTC检测限和定量限分别为5.0、10.0μg/kg。ECTC检测限和定量限分别为20.0、30.0μg/kg。CTC和ECTC在肉鸡肌肉中平均回收率分别在73.0%~95.20%和81.80%~101.60%,相对标准偏差(RSD)分别在8.85%~12.18%和2.05%~9.29%;在肉鸡肝脏中平均回收率分别在67.06%~90.80%和78.64%~106.98%,RSD分别在2.90%~8.88%和4.72%~9.48%;肉鸡肌肉和肝脏中CTC、代谢物ECTC的残留量分别在给药后第1、5 d时达到最高。肉鸡肝脏中ECTC的残留量约占总残留量的1/2。停药后CTC及其代谢物ECTC代谢缓慢,在停药第6 d时,仍能检测到CTC及其代谢物ECTC,但其残留量均低于最高残留限量(MRLs)。从保障食品安全方面考虑,建议我国将CTC和ECTC均作为金霉素的残留标示物进行监测。     

Cardiopulmonary effects of reverse Trendelenburg position at 5° and 10° in sevoflurane-anesthetized steers

Objective

To assess the cardiopulmonary effects caused by reverse Trendelenburg position (RTP) at 5° and 10° in sevoflurane-anesthetized yearling steers.

前列腺素E2和F2α对LPS刺激后奶牛子宫内膜上皮细胞COX-1和COX-2表达的影响

试验旨在阐明前列腺素E₂（prostaglandin E₂,PGE₂）和F_2α（prostaglandin F_2α,PGF_2α）对体外培养的奶牛子宫内膜上皮细胞中环氧合酶-1（cyclooxygenase-1,COX-1））与环氧合酶-2（cyclooxygenase-2,COX-2）表达的影响。培养奶牛子宫内膜上皮原代细胞和传代细胞,第4代细胞以1×10⁶个/孔接种于6孔板,以10^-7mol/L PGE₂和PGF_2α分别预处理细胞24 h,以100 ng/mL细菌脂多糖（lipopolysaccharides,LPS）刺激细胞4、8和12 h后分别提取RNA和总蛋白质,采用实时荧光定量PCR与Western blotting等技术检测COX-1与COX-2 mRNA和蛋白质的表达量。结果表明,与对照组相比,COX-1 mRNA表达量在PGE₂单独作用4、8和12 h后显著上调（P<0.05）;COX-2 mRNA表达量在PGE₂单独作用4和12 h后显著上调（P<0.05）,PGE₂单独处理使COX-1、COX-2蛋白表达量均显著上调（P<0.05）。与对照组相比,LPS刺激8和12 h时COX-1 mRNA表达量显著下调（P<0.05）,LPS刺激后COX-1蛋白表达量无显著变化（P>0.05）;LPS刺激后4、8和12 h时COX-2 mRNA表达量显著上调（P<0.05）,LPS刺激后COX-2蛋白表达量显著上调（P<0.05）。与LPS单独处理组相比,LPS+PGE₂处理组在8和12 h时COX-1和COX-2 mRNA表达量均显著上调（P<0.05）,同时COX-1和COX-2蛋白表达量也显著上调（P<0.05）。PGF_2α在LPS未刺激和刺激后对COX-1和COX-2 mRNA的表达无显著影响（P>0.05）,仅在PGF_2α单独处理8和12 h后COX-1 mRNA表达量上调（P<0.05）。两种激素联合处理与各自单独处理及LPS单独刺激相比,对COX-1和COX-2 mRNA表达具有一定的协同诱导作用。     

牛TLR4基因的遗传多态性与乳房炎的关联分析

TLR4通过识别病原体而激活免疫细胞，在先天免疫和适应性免疫防御中起着重要作用。以中国荷斯坦奶牛、三河牛和中国西门塔尔牛共397头为研究对象，利用创造酶切位点PCR法扩增243bp的目的片段，通过限制性内切酶HinfⅠ酶切来检测TLR4第3外显子的多态性，结果发现扩增产物的27bp处C到T的突变使得多态位点产生，编码的氨基酸由苏氨酸变为异亮氨酸。A、B2个等位基因在3个群体中均有分布，A等位基因占优势（大于78％），经χ^2适合性检验，三河牛在该位点未达到Hardy-Weinberg平衡状态（P〈0．05）。运用SAS8．2软件采用最小二乘法拟合线性模型，将该基因座不同基因型与奶牛乳房炎进行了关联分析，结果表明：AA基因型为乳房炎抗性基因型（P〈0．05），A等位基因为乳房炎抗性的有利基因。     

奶牛乳房炎金黄色葡萄球菌8种毒力基因的PCR检测

为研究奶牛乳房炎金黄色葡萄球菌(Staphylococcus aureus,SA)主要毒力因子的分布情况,本试验利用PCR对70株临床型乳房炎、55株隐性乳房炎金黄色葡萄球菌的8种主要毒力基因进行检测。结果显示,nuc、ClfA、TSST-1、PVL、Hla、Hlb、FnBPA和FnBPB 8种毒力基因在临床型乳房炎SA菌株中的检出率分别为:100.0%(70株)、100.0%(70株)、0(0株)、5.7%(4株)、100.0%(70株)、11.4%(8株)、97.1%(68株)和100.0%(70株);在隐性乳房炎SA菌株中的检出率分别为:100.0%(55株)、100.0%(55株)、0(0株)、70.9%(39株)、98.2%(54株)、9.1%(5株)、100.0%(55株)和100.0%(55株)。结果表明,nuc、ClfA、Hla、Hlb和FnBPA 5种毒力基因是引起奶牛乳房炎的SA的最主要毒力基因;PVL基因可能是引起隐性乳房炎的重要致病基因。     

牛基因组DNA两种提取方法的比较研究

研究哺乳动物基因组DNA的水煮抽提法，并将传统的酚仿抽提法和水煮法进行比较，水煮法即通过对细胞的煮沸和冷却，使细胞破裂、蛋白变性，从而获得用于PCR扩增的模板DNA的方法。通过电泳分析和PCR扩增检测了所提取基因组DNA的完整性、可行性。检测时采用3对引物对2种方法提取的DNA进行了PCR扩增，同时对冷冻保存的基因组DNA也进行了扩增检测。结果表明：水煮法提取基因组DNA是一种快速、简便、经济、高效的方法。     

Aflatoxin B<Subscript>1</Subscript> Binding to Sorbents in Bovine Ruminal Fluid

A recent approach to the problem of contamination of agricultural products by aflatoxin B₁ (AFB₁) is to add non-nutritional adsorbents to animal diets in order to sequester ingested aflatoxins. We conducted in vitro experiments to develop a rapid and cheap model using ruminal fluid to assess the ability of sorbent materials to bind AFB₁. Seven sorbents (hydrated sodium calcium aluminosilicate; clinoptilolite; zeolite; two types of bentonite; sepiolite; and PHIL 75), commonly added to bovine diets were incubated in water and ruminal fluid in the presence of AFB₁. Hydrated sodium calcium aluminosilicate, sepiolite and one of the bentonites bound 100% of the AFB₁ in the presence of both ruminal fluid and water; clinoptilolite bound about 80% of AFB₁ in both liquids; whereas the affinities for the mycotoxin of zeolite (50%) and the other sample of bentonite (60%) in water seem to be increased by about 40% in ruminal fluid incubations. PHIL 75 had the poorest binding ability: about 30% in water and 45% in ruminal fluid. In view of the differences in toxin binding in water and ruminal fluid, it is preferable to use the ruminal fluid model for the in vitro pre-screening of sorbent materials potentially useful as adjuvants to ruminant feeds.