Pharmacokinetics of Flunixin after Intravenous Administration in Healthy and Endotoxaemic Rabbits

The pharmacokinetics of flunixin were determined after intravenous bolus injection at a single dose (2.2 mg/kg) in healthy rabbits and diseased rabbits with Escherichia coli lipopolysaccharide-induced septic shock. Six adult New Zealand White rabbits were used. Concentrations of drug in plasma were determined by HPLC. Pharmacokinetics were best described by a two-compartment open model. In healthy rabbits, there was a high plasma clearance (0.62 L/(h kg)), and a relatively short elimination half-life (1.19 h). In endotoxaemic rabbits, total plasma clearance (0.43 L/(h kg)) was significantly lower (p<0.05), and elimination half-life (1.90 h) and AUC₀-∞ (5.29 (μg h)/ml) were significantly higher (p<0.05) than in healthy animals. The changes of pharmacokinetics of flunixin in rabbits with septic shock could be of clinical significance, and may require monitoring of plasma flunixin levels in endotoxaemic status.     

乳头灌注脂多糖对奶牛乳腺组织及机体免疫的影响

旨在通过乳头灌注脂多糖(LPS)体外模拟奶牛急性乳腺炎,研究奶牛乳腺组织及机体免疫活化状态。选用4头泌乳后期的荷斯坦奶牛16个乳区为研究对象,采用交叉试验设计,每期试验2 d,试验间隔10 d,试验组乳头管乳头灌注LPS 0. 1 mg,对照组乳头管灌注等量生理盐水。结果:试验组灌注LPS后,奶牛体温值和乳汁中体细胞数随时间变化而逐渐升高,而奶牛血液中白细胞数随时间变化而逐渐降低;乳腺组织的腺泡腔内充斥大量炎性细胞,部分腺泡组织萎缩甚至崩塌;试验组灌注LPS 12 h后奶牛血清中的髓过氧化物(MPO)活性极显著下降(P<0. 01),而乳腺组织中的MPO活性则极显著上升(P<0. 01);奶牛血清及乳清中炎症因子肿瘤坏死因子(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)的蛋白含量明显上升(P<0. 01);乳腺组织中Toll样受体4 (TLR4)、核因子NF-κB (p65)蛋白的表达量相比对照组极显著上升(P<0. 01)。研究表明,LPS能够诱导奶牛发生乳腺炎,使乳腺组织发生损伤,并激活机体的TLR4/NF-κB信号通路进而促进其下游炎症因子TNF-α、IL-1β、IL-6的基因的表达,使机体发生免疫应答。     

Evaluation of the ability of colistin,amoxicillin (components of Potencil®), and fluoroquinolones to attenuate bacterial endotoxin‐ and Shiga exotoxin‐mediated cytotoxicity—In vitro studies

Escherichia coli is one of the major pathogens in humans and animals causing localized and systemic infections, which often lead to acute inflammation, watery diarrhea, and hemorrhagic colitis. Bacterial lipopolysaccharide (LPS) and Shiga exotoxins (Stx) are mostly responsible for such clinical signs. Therefore, highly effective treatment of E. coli infections should include both eradication of bacteria and neutralization of their toxins. Here, for the first time, we compared the in vitro ability of common antibiotics to decrease LPS‐ and Stx‐mediated cytotoxicity: colistin, amoxicillin (used separately or combined), enrofloxacin, and its metabolite ciprofloxacin. Three experimental scenarios were realized as follows: (a) the direct effect of antibiotics on endotoxin, (b) the effect of antibiotic treatment on LPS‐mediated cytotoxicity in an experiment mimicking “natural infection,” (c) the effect of antibiotics to decrease Stx2e‐mediated cytotoxicity. Two cell lines, A549 and Vero cells, were used to perform cytotoxic assays with the methyl tetrazolium (MTT) and lactate dehydrogenase leakage (LDH) methods, respectively. Colistin and amoxicillin, especially used in combination, were able to attenuate LPS toxic effect, which was reflected by increase in A549 cell viability. In comparison with other antibiotics, the combination of colistin and amoxicillin exhibited the highest boster or additive effect in protecting cells against LPS‐ and Stx2e‐induced toxicity. In summary, in comparison with fluoroquinolones, the combination of colistin and amoxicillin at concentrations similar to those achieved in plasma of treated animals exhibited the highest ability to attenuate LPS‐ and Stx2e‐mediated cytotoxicity.     

猪胸膜肺炎放线杆菌单克隆抗体的制备与鉴定

目的:建立抗血清1型猪胸膜肺炎放线杆菌(APP-1)脂多糖(LPS)的单克隆抗体杂交瘤细胞株,为今后建立该菌的免疫检测技术奠定基础。方法:腹腔注射APP-1标准株的甲醛固定抗原免疫BALB/c小鼠,ELISA检测血清中LPS抗体滴度并选择值最高的小鼠用于细胞融合,且于融合前3天加强免疫1次。常规方法进行融合,HT、HAT培养液选择培养融合后的细胞,ELISA检测细胞培养上清中的LPS抗体,阳性杂交瘤细胞用有限稀释法克隆传代。结果:获得2株稳定分泌抗APP-1LPS抗原的单克隆抗体杂交瘤细胞株,其小鼠腹水单克隆抗体效价132000～164000。结论:已成功建立2株稳定分泌抗APP-1LPS抗原的单克隆抗体杂交瘤细胞株,为今后建立该菌的免疫胶体金分型诊断技术奠定基础。     

Apparent Seroprevalence of Salmonella spp. in Harp Seals in the Greenland Sea as Determined by Enzyme-linked Immunosorbent Assay

An indirect ELISA was developed as a possible tool for surveillance of the seroprevalence of Salmonella spp. in harp seals. This species is hunted for human consumption and thus transmission of disease to humans cannot be excluded. To cover a broad spectrum of serogroups, a mixture of the lipopolysaccharides (LPS) of S. typhimurium and S. choleraesuis was used as the antigen in this pilot study. Chicken anti-harp-seal immunoglobulin horseradish peroxidase conjugate served as the immunoconjugate. Sera from four captive harp seals, which were Salmonella culture-negative and had no clinical or historical evidence of salmonellosis, were used as negative controls. After immunization with an inactivated S. typhimurium vaccine, further sera from these seals were used as positive controls, as no serum from naturally infected animals was available. Serum samples from 93 harp seals caught in the Greenland sea in 1999 were examined, and anti-Salmonella antibodies were found in the samples from two individuals (seroprevalence 2.2%). Although Salmonella has been isolated from other pinniped species, this is the first documentation of Salmonella-seropositive harp seals. This study contributes to the evaluation of the importance of salmonellosis in arctic marine mammals and thus to the prevention of potential outbreaks of this important zoonosis.     

Pseudomonas for biological control of dutch elm disease. I. Labeling,detection and identification of pseudomonas isolates injected into elms; comparison of various methods

To understand the mechanisms involved in biological control of Dutch elm disease byPseudomonas, data were needed on the distribution of the introduced bacteria within elm and on the development of the bacterial population over a period of time.As traditional biochemical identification techniques are not suitable for distinguishment between individualPseudomonas isolates, three alternative approaches were compared.

脂多糖应激对添加不同锌源蛋鸡免疫反应和组织锌代谢的影响

36只海兰褐壳蛋鸡按2×3因子进行试验,饲喂3种日粮(分别为基础日粮、基础日粮中添加60 mg/kg硫酸盐和氨基酸络合形式的锌),每组随机选取半数腹腔注射脂多糖(LPS),另一半注射同样剂量的生理盐水,研究LPS应激对饲喂不同锌源蛋鸡产蛋性能、直肠温度、血清IL 1β以及肝脏和脾脏组织MT和锌含量的影响。结果表明,日粮锌源以及锌源与LPS互作对蛋鸡的产蛋率、蛋重和破蛋率均没有显著影响(P> 0.05),LPS攻毒显著降低产蛋率(P< 0.05)并增加破蛋率(P< 0.05),其中添加氨基酸络合锌(ZnAA)蛋鸡的产蛋率显著(P< 0.01)低于基础日粮组;饲喂ZnAA的蛋鸡LPS攻毒后直肠温度峰值出现以及发热消退的时间均早于添加ZnSO4和不添加锌的蛋鸡。饲喂ZnAA的蛋鸡注射LPS 3 h后血清IL 1β的水平显著高于饲喂ZnSO4 的蛋鸡,而注射LPS 12 h后血清IL 1的水平则显著低于硫酸锌组(P< 0.05)。与ZnSO4 相比,添加ZnAA具有提高注射LPS后肝脏和脾脏组织锌浓度的趋势(P< 0.09)。锌源和LPS应激以及两者的互作显著影响组织金属硫因(MT)的含量(P< 0.001),在LPS应激蛋鸡中,添加ZnAA的蛋鸡肝脏和脾脏MT的水平显著高于添加ZnSO4 的蛋鸡(P< 0 001):与饲喂基础日粮的蛋鸡相比,添加ZnAA蛋鸡肝脏和脾脏MT浓度分别提高266%和225%,而添加ZnSO4仅提高158%和151%。由     

LPS对大鼠肠黏膜微血管内皮细胞分泌NO的影响

一氧化氮（Nitric oxide，NO）是一种由血管内皮细胞分泌的内源性血管舒张因子。NO作为一种重要的信使分子具有舒张血管、神经传递以及杀灭微生物和肿瘤细胞等多种生物学功能。     

脂多糖应激对断奶仔猪肠黏膜免疫屏障的影响

选用12头(21±1)d断奶仔猪,随机分为2个处理,研究脂多糖(LPS)刺激对肠黏膜免疫屏障功能的影响。禁食12h后,LPS组仔猪腹膜注射100μg/kgBW LPS,对照组注射等量的生理盐水。48h后,屠宰仔猪,打开腹腔,取小肠组织样品,测定小肠上皮间淋巴细胞、杯状细胞、肥大细胞数、肠集合淋巴结增殖、凋亡细胞数。结果表明:LPS刺激显著降低十二指肠、回肠上皮间淋巴细胞数(P<0.01),极显著增加小肠各段肥大细胞数(P<0.01);LPS刺激显著增加回肠集合淋巴结凋亡细胞数(P<0.05),但对增殖细胞数没有影响(P>0.05);LPS刺激可增加仔猪小肠杯状细胞数,但两处理组间差异不显著(P>0.05)。这些结果显示,LPS应激可导致肠黏膜免疫屏障功能改变,进而加重机体出现急性感染症状。     

两种组胺受体拮抗剂对内毒素作用下雏鸡体温的影响

为了研究组胺H1、H2受体拮抗剂对内毒素攻毒后雏鸡体温的影响,探讨内源性组胺在内毒素攻毒时导致雏鸡发热的作用机理。采用100羽雏鸡随机分5组,即模型组、扑尔敏组、西咪替丁组、扑和西联合组和空白组。空白组中午腹腔注射生理盐水,其余各组注射等剂量的LPS,早、晚肌注相应药物。实验持续15 d,分别于攻毒后0 d、5 d、10 d、15 d,各组随机抽10个样本。在注射LPS后0 h、2 h、4 h6、h,测定雏鸡直肠温度,结果:(1)急性实验中,扑尔敏组2 h、4 h体温显著降低(P<0.05);而西咪替丁组和扑和西联合组在2h时体温极显著降低(P<0.01)。(2)慢性实验中,0 d5、d体温无显著差异;10 d、15 d扑尔敏组体温显著降低(P<0.05);15 d扑尔敏组、扑和西联合组体温显著降低(P<0.05)。结果表明:在内毒素引起的炎症反应中,内源性组胺主要是通过H1受体介导体温升高。