Monitoring and identification of residues of anabolic preparations in slaughtered cattle by HPLC with diode array detection

Summary

The new combination of isocratic high performance liquid chromatography (HPLC) with on line UV spectrum detection via a diode array configuration has been applied to the detection and identification of anabolics present in application sites of cattle.

Combination of the characteristic retention time in the HPLC chromatogram and a comparison of the full spectrum between 190–400 nm of the anabolic components with that of a standard resulted in a very reliable identification. By means of this method 117 samples of application sites were investigated for the presence of anabolic residues. Of the xenobiotic anabolics, 19‐nortestosterone (NT) was found most frequently (in 96 cases), whereas diethylstilbestrol (DES) was found in only 11 cases.

In all samples the identification of NT and DES was confirmed by high resolution gas chromatography‐mass spectrometry (GCMS).     

淹水对紫花苜蓿南北方品种抗氧化酶和无氧呼吸酶的影响

以紫花苜蓿南方育成品种渝苜1号和北方地方品种新疆紫泥泉为材料,在幼苗长出4片真叶时进行淹水处理,在淹水的第10天测定植株的生物量,并分别在淹水的第0,2,4,6,8,10天测定叶片中的丙二醛(MDA)含量以及叶片抗氧化酶和根系无氧呼吸酶的活性,以探明两个品种对淹水胁迫的耐受性差异及其生理响应机制。结果表明,淹水胁迫使紫花苜蓿的生物量减少,但与新疆紫泥泉相比,渝苜1号生物量的降幅小。淹水时渝苜1号叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)等抗氧化酶活性比新疆紫泥泉的高,而叶片MDA含量比新疆紫泥泉的低;根系无氧呼吸酶活性在淹水胁迫中均增强,但渝苜1号的乙醇脱氢酶(ADH)和丙酮酸脱羧酶(PDC)活性增加明显,乙醇发酵更强;而新疆紫泥泉的乳酸脱氢酶(LDH)的活性增加更为突出,乳酸发酵强。淹水影响紫花苜蓿正常生长,但南方品种渝苜1号比北方品种新疆紫泥泉对淹水胁迫更具耐受性,因为前者具有比较高的抗氧化酶活性和比较强的乙醇发酵途径,有利于增强植株抗淹水胁迫能力。     

高致病性猪繁殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析

为了弄清2006年在我国南方一些猪场暴发的一种以高热、高发病率和高死亡率为特征的传染病的病因,我们从12个省市48个发病猪场采集临床样品进行了猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)的分离鉴定和分子流行病学分析。结果显示从48个猪场样品中均检测出了PRRSV,通过完整的ORF5和部分Nsp2基因序列比较分析发现来自12个省市不同猪场的PRRSV高度同源,而且在Nsp2基因上都有两个相同位点的缺失;从分离的15株PRRSV中选择湖南分离株HuN4人工感染5头60日龄的健康猪,结果5头猪分别在7 d～21 d内全部死亡,感染猪的临床症状和现地所见十分相似。本次试验结果表明新出现的以Nsp2基因部分缺失为特征的PRRSV对猪是高致病性的,可能是2006年大部分猪场暴发"高热综合症"的主要病因。     

褪黑素对内毒素血症山羊肝细胞线粒体呼吸功能的影响

将48只健康山羊随机分为4组：对照组（A组）、内毒素（LPS）组（B组）、内毒素＋褪黑素（LPS＋MT）组（C组）、褪黑素（MT）组（D组）,在处理后第3 h和6 h提取肝细胞线粒体,采用Clark氧电极技术测定线粒体呼吸控制率（RCR）、磷氧比值（P/O）、氧化磷酸化效率（OPR）的变化,探讨了MT对LPS诱导的山羊内毒素血症机体线粒体呼吸功能的影响。结果显示,内毒素组山羊在第3 h和6 h的RCR、P/O、OPR显著低于对照组（P〈0.05）;应用褪黑素后,即内毒素＋褪黑素组第3 h的肝细胞线粒体RCR、P/O、OPR明显升高（P〈0.05）;褪黑素组第3 h的RCR明显高于对照组（P〈0.05）,而褪黑素组第3 h的P/O、OPR与对照组差异不显著（P〉0.05）。证实,山羊内毒素血症时线粒体的呼吸功能明显下降,褪黑素对肝细胞线粒体呼吸功能具有一定的保护作用。     

Acute respiratory distress syndrome in dogs and cats: a review of clinical findings and pathophysiology

Objective: To review the clinical and pathophysiologic aspects of acute respiratory distress syndrome (ARDS) in dogs and cats. Data sources: Data from human and veterinary literature were reviewed through Medline and CAB as well as manual search of references listed in articles pertaining to acute lung injury (ALI)/ARDS. Human data synthesis: Since the term ARDS was first coined in 1967, there has been a abundance of literature pertaining to this devastating syndrome in human medicine. More complete understanding of the complex interactions between inflammatory cells, soluble mediators (e.g., tumor necrosis factor, interleukin (IL)‐6, IL‐8, platelet activating factor) and the clinical patient has provided for timely recognition and mechanistically based protective strategies decreasing morbidity and mortality in human patients with ARDS. Veterinary data synthesis: Although little is known, ARDS is becoming a more commonly recognized sequela in small animals. Initial case reports and retrospective studies have provided basic clinical characterization of ARDS in dogs and cats. Additionally, information from experimental models has expanded our understanding of the inflammatory mechanisms involved. It appears that the inflammatory processes and pathologic changes associated with ARDS are similar in dogs, cats, and humans. Conclusions: Unfortunately, current mortality rates for ARDS in small animals are close to 100%. As our capability to treat patients with advanced life‐threatening disease increases, it is vital that we develop a familiarity with the pathogenesis of ARDS. Understanding the complex inflammatory interactions is essential for determining effective preventative and management strategies as well as designing novel therapies for veterinary patients.     

Bold fusion: transtracheal wash from a Jersey calf

A 10-week-old, female, Jersey calf was referred to the University of Minnesota Veterinary Medical Center for evaluation of difficult breathing and inappetence of 18-hours duration. Cytologic examination of a transtracheal wash specimen revealed pyogranulomatous inflammation, with increased numbers of multinucleated cells. Differential diagnoses included established bacterial infection, viral infection, fungal infection, and foreign body reaction. Immunocytochemical staining was done on a direct smear of the transtracheal wash using a cocktail of 4 mouse monoclonal antibodies against 3 human respiratory syncytial virus proteins. Strong intracytoplasmic staining within mononuclear cells was observed and a diagnosis of bovine respiratory syncytial virus infection was made. Immunocytochemistry may be used as a rapid, inexpensive, adjunctive diagnostic tool for the antemortem diagnosis of bovine respiratory syncytial virus on transtracheal wash specimens.     

猪繁殖与呼吸综合征病毒A06株单克隆抗体的制备及鉴定

用纯化的猪繁殖与呼吸综合征病毒(PRRSV)A06株病毒抗原免疫Balb/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合.经间接ELISA方法筛选,有限稀释法克隆,获得4株稳定分泌特异性单克隆抗体的杂交瘤细胞株,分别命名为3D12、4E10、6E2和7G6.其细胞培养上清效价分别为1:256、1:512、1:512和1:256,小鼠腹水效价分别为1:1.024×106、1:2.048×106、1:1.024×106和1:2.048×106.亚型鉴定表明,3D12、4E10、6E2和7G6分别为IgG2b、IgG2b、IgG1和IgG2a.ELISA检测结果显示,4种PRRSV单克隆抗体仅与PRRSV反应,不与其他病毒反应,表明均为抗PRRSV的型特异性单克隆抗体.对单克隆抗体腹水液进行IgG提取纯化,SDS-PAGE电泳验证,仅出现两条清晰条带,无杂带出现,证明纯化效果较好.这4种单克隆抗体的获得,为建立PRRSV检测方法提供了强有力的工具.     

结缕草种子打破休眠过程呼吸途径的研究

采用呼吸抑制剂测定了结缕草休眠与打破休眠种子在发芽过程中的各个呼吸代谢途径－糖酵解呼吸途径（ＥＭＰ）、三羧酸循环呼吸途径（ＴＣＡ）、磷酸戊糖呼吸途径（ＰＰＰ）的呼吸速率及其占总呼吸速率的比例，结果表明，休眠种子的总呼吸速率显著低于打破休眠种子的总呼吸速率，造成这种差异的原因是磷酸戊糖途径及三羧酸循环途径的呼吸速率低，这是休眠结缕草种子不能发芽的原因之一。     

不同毒力猪繁殖与呼吸综合征病毒对仔猪肺和外周免疫器官损伤的研究

为研究不同毒力的PRRSV对仔猪肺脏和外周免疫器官损伤的差异,本实验分别采用PRRSV变异株(HuN4株)和PRRSV经典株(CH-1a株)感染35日龄健康的断奶仔猪,并在感染后0 d、3 d、7 d、10 d和14 d各迫杀3头,检测肺、颌下淋巴结、肠系淋巴结、腹股沟淋巴结、扁桃体和脾脏的病毒载量及病理变化情况,同时检测血清中抗PRRSV的抗体水平。结果表明:感染后3 d肺脏及各免疫器官可检测到病毒,HuN4感染组病毒载量比CH-1a感染组病毒载量高1 000倍;HuN4感染组病毒载量峰值出现在感染后10 d,而CH-1a感染组维持着较低水平的病毒载量。组织病理学检测显示HuN4感染组淋巴结内淋巴细胞显著减少,呈空泡状;CH-1a感染组淋巴结内淋巴细胞轻度减少,呈星隙状。本实验表明HuN4株比CH-1a株对肺和外周免疫器官造成更严重的损伤。     

Characterization of an intravenous lipopolysaccharide inflammation model in calves with respect to the acute-phase response

Our objective was to develop a lipopolysaccharide (LPS) inflammation model in calves to evaluate the acute-phase response with respect to the release of pro-inflammatory cytokines and acute-phase proteins, fever development and sickness behaviour. Fourteen 4-week-old male Holstein Friesian calves were included and randomly assigned to a negative control group (n = 3) and an LPS-challenged group (n = 11). The latter received an intravenous bolus injection of 0.5 μg of LPS/kg body weight. Blood collection and clinical scoring were performed at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 8, 12, 18, 24, 28, 32, 48, 54 and 72 h post LPS administration (p.a.). In the LPS group, the following clinical signs were observed successively: tachypnoea (on average 18 min p.a.), decubitus (29 min p.a.), general depression (1.75 h p.a.), fever (5 h p.a.) and tachycardia (5 h p.a.). Subsequent to the recovery from respiratory distress, general depression was prominent, which deteriorated when fever increased. One animal did not survive LPS administration, whereas the other animals recovered on average within 6.1 h p.a. Moreover, the challenge significantly increased plasma concentrations of tumour necrosis factor-α, interleukin 6, serum amyloid A and haptoglobin, with peaking levels at 1, 3.5, 24 and 18 h p.a., respectively. The present LPS model was practical and reproducible, caused obvious clinical signs related to endotoxemia and a marked change in the studied inflammatory mediators, making it a suitable model to study the immunomodulatory properties of drugs in future research.