鸭传染性浆膜炎实验性感染方法的研究

采用气管、腹腔、嗉囊及脚蹼四种方式注射鸭疫巴氏杆菌实验性感染鸭传染性浆膜炎 ,以确立最佳感染途径 ,并在此基础上探索其理想的感染菌量 ,结果表明 :以 0 .15～ 0 .2 0ml(3× 10 8cfu/ml)肉汤培养物脚蹼内注射获得理想的实验感染效果。     

鸭疫里氏杆菌生长曲线的测定

采用P L琼脂平板涂布法 ,测定鸭疫里氏杆菌WJ4株 (血清 1型 )、FXb7株 (血清 2型 )在P L肉汤中静置培养和振荡培养情况下的生长曲线。结果表明 ,在P L肉汤中振荡培养时 ,WJ4株培养 10h达生长最高峰 ( 4 7× 10 1 0 CFU mL) ,FXb7株 11h活菌数最高达 ( 4 2× 10 1 0 CFU mL) ;在静置培养时 ,WJ4株培养 2 2h达最高峰 ( 1 8× 10 9CFU mL) ,FXb7株 2 3h可达最高峰 ( 1 3× 10 9CFU mL)。     

嗜水气单胞菌GYK1株培养基的优化

在营养肉汤培养基的基础上,经过一系列嗜水气单胞菌GYK1株培养试验,结果证明,成本价格较低的蔗糖、酵母膏可以代替葡萄糖、牛肉膏,从而降低培养基原料成本;添加少量的K2HPO4和微量元素溶液(Mg2+、Fe2+、Zn2+、Mn2+)可明显提高培养液的活菌数;通过正交试验得到低成本、高得率的适合嗜水气单胞菌GYK1菌株生长的培养基配方,该培养基组成(g/L):蛋白胨10.0、酵母膏10.0、蔗糖15.0、K2HPO42.28;NaCl 5.0微量元素(硫酸镁0.1,硫酸亚铁0.04,硫酸锌0.00375,二氯化锰0.0021)。24 h摇瓶培养,菌液中的最高活菌数可达316×108cfu/mL,比营养肉汤培养基活菌数(151×108cfu/mL)提高100%以上。     

大肠杆菌对接种新城疫疫苗后禽体的影响

俄学者杜尔德叶夫做了两个试验研究了大肠杆菌(E.coli)对接种新城疫疫苗雏鸡的影响。第一个试验是取10日龄雏鸡(n=35)分两组(试验与对照)。两组雏鸡均在10和20日龄两次接种了新城疫疫苗各0.2ml。试验组雏鸡(n=20)于10日龄时腹腔内注入含有10~9 M.T.(LD_(50))E.coli一日肉汤培养物0.25ml。对照组(n=15)不予感染。第二个试验是取     

一株产肠毒素金黄色葡萄球菌致死小鼠的初步研究

目的:观察产肠毒素金黄色葡萄球菌对小鼠的致病性。方法:用一株肠毒素检测阳性的金黄色葡萄球菌(SEF101)24h肉汤培养物,每只0.5mL肌肉、腹腔或尾静脉注射染菌。结果:金黄色葡萄球菌(SEF101)24h肉汤培养物经肌肉、腹腔或尾静脉注射染菌均可致小鼠死亡,最小感染剂量为每只鼠腹腔注射菌量3.3×105CFU即可引起小鼠感染死亡,24h培养物经80℃30min灭活处理仍可致小鼠死亡,经VIDAS法检测,24h肉汤培养物均呈肠毒素阳性。结论:金黄色葡萄球菌(SEF101)24h肉汤培养物染菌量3.3×105CFU即可引起小鼠感染死亡,金黄色葡萄球菌(SEF101)为产肠毒素菌株。     

酵母培养物水溶物对离体草鱼肠道黏膜细胞生长及细胞膜完整性的影响

本试验以酵母培养物水溶物为试验材料,添加于离体草鱼肠道黏膜细胞培养液中,研究酵母培养物水溶物在不同浓度、不同作用时间下对细胞生长及细胞膜完整性的影响。采用单因子试验设计,设1个对照组及5个酵母培养物水溶物组(YC-1~5组),各组均设96个重复,每个重复为1个培养孔。对照组的培养液中不添加酵母培养物水溶物,YC-1~5组的培养液中酵母培养物水溶物的浓度分别为10、25、50、100、200 mg/L。结果表明:培养液中添加50~200 mg/L酵母培养物水溶物对细胞形态无损伤,100~200 mg/L酵母培养物水溶物在添加后3 h时显著增强细胞活性(P<0.05),50 mg/L酵母培养物水溶物在添加后6 h时显著增强细胞活性(P<0.05)。与对照组相比,3 h时各酵母培养物水溶物组培养液中乳酶脱氢酶(LDH)活力均没有显著变化(P>0.05),6 h时YC-4、YC-5组显著降低(P<0.05),9 h时YC-3、YC-4、YC-5组显著降低(P<0.05),12 h时YC-2、YC-3、YC-4、YC-5组显著降低(P<0.05)。各时间点(3、6、9、12 h)各酵母培养物水溶物组培养液中谷丙转氨酶(GPT)、谷草转氨酶(GOT)活力与对照组相比均没有显著变化(P>0.05),但在6~9 h时GPT活力均低于对照组。由此得出,培养液中添加10~200 mg/L酵母培养物水溶物能促进离体草鱼肠道黏膜细胞的生长,保护细胞膜的完整性,其发挥保护作用的适宜浓度为50~200 mg/L。     

酵母培养物对育肥猪生产性能和消化率的影响

酵母培养物是一种营养丰富,产量大,成本低,饲用安全的优质饲料原料,试验选择80头体质量相近的健康(杜×长×大)三元杂交育肥猪。采用单因素完全随机试验设计,将80头育肥猪随机分为2组,每组4个重复,每重复10头猪采用自动喂料系统饲喂,记录每头猪的生产性能。通过对试验组饲喂基础日粮替换10%的酵母培养物,对照组饲喂基础日粮,研究酵母培养物对育肥猪生产性能的影响。结果表明:酵母培养物可提高生长育肥猪的生产性能及消化率,提高育肥猪日增质量,降低料肉比,从而可提高猪场效益。     

奶牛源表皮葡萄球菌的分离鉴定

为了探讨生牛乳中表皮葡萄球菌的分离鉴定方法,对某奶牛场的生鲜乳进行细菌分离培养和生化试验鉴定。结果显示：使用氯化钠肉汤（氯化钠质量分数为7.5%）增菌,使用Baird-Parker琼脂培养基选择培养,使用营养琼脂培养基纯化培养,可获得表皮葡萄球菌分离株;使用vitek2compact全自动微生物分析系统鉴定表皮葡萄球菌分离株,可信水平为良好;本实验获得的表皮葡萄球菌分离株与经典表皮葡萄球菌模式菌株符合率为93%。     

应用酶标抗体染色法实验检定空肠弯曲菌

应用酶标抗体染色法检定空肠弯曲菌,具有较高的特异性及敏感性,除金黄色葡萄球菌呈现轻度着染外(可以从形态学上排除),对沙门氏菌等9种杂菌均不发生非特异性染色反应;用以检查空肠弯曲菌纯培养物,敏感性可达400万个菌/ml。对同时接种10个空肠弯曲菌和1亿个杂菌的30h选择性增菌肉汤培养物,酶染色法可准确地检出,而且不受杂菌的干扰。检查47份鸡粪便标本和38株猪源空肠弯曲菌,其结果与常规细菌分离鉴定结果相同。建立的检定空肠弯曲菌的酶标抗体染色法可在2h内完成。     

腐蹄病C型节瘤拟杆菌纤毛蛋白基因T程疫苗对家兔的免疫试验

将转化C型节瘤拟杆菌纤毛蛋白基因表达子粒的工程菌PAK/2pfsC在含羧苄青霉素营养肉汤培养基中培养,用MgCl2法在培养物上清中提取表达产物(纤毛蛋白).将表达的纤毛蛋白产物用弗氏完全佐剂乳化,制成疫苗.用疫苗免疫3只健康家兔,21 d后再次接种;定期采血,用对流免疫电泳和K凝集实验检测试验家兔的体液免疫应答及抗体水平.结果发现,免疫7 d即可产生相应抗体,21 d后抗体效价达到8 000×以上,而且高滴度抗体可维持6个月以上.试验表明,腐蹄病C型节瘤拟杆菌纤毛蛋白基因工程疫苗具有较好的免疫应答和免疫原性.     

Comparison of the effects of Sesbania drummondii on the hepatic microsomal monooxygenase systems of chickens and rats

Sesbania drummondii, a toxic leguminous shrub found throughout the southeastern United States, induces different responses in chicken vs rat hepatic microsomal monooxygenase systems. Groups of 4- to 8-week-old Sprague-Dawley rats and White Leghorn chickens were given extracts of S drummondii by gavage for 3 days. Doses, which were 0.4 and 0.8% of daily body weights, respectively, for the rats and chickens, were adjusted to induce similar clinical lesions in the 2 species. The hepatic microsomal monooxygenase systems of control and treated animals were compared, using cytochrome P-450 content, cytochrome b5 content, NADH- and NADPH-cytochrome c-reductase activity, and 6 cytochrome P-450 mediated enzyme activities. Increases of twofold in the cytochrome P-450 content, NADPH-cytochrome c-reductase, aminopyrine-N-demethylase, aniline hydroxylase, ethoxycoumarin-O-deethylase, and aryl hydrocarbon hydroxylase activities; fourfold in the aldrin epoxidase activity; and 15-fold in the ethoxyresorufin-O-deethylase activity were observed in the S drummondii-treated chickens. In contrast, the treated rats had nearly twofold decreases in these values, suggesting a species-specific effect of S drummondii on microsomal monooxygenase systems, ie, induced with S drummondii.     

槲皮素对Aroclor1254诱导的孕鼠肝细胞CYP450表达的影响

采用胶原酶二步灌流法获取怀孕大鼠的原代肝细胞,以Aroclor1254诱导肝细胞损伤,用不同剂量的槲皮素分别处理损伤的肝细胞24～72h,RT—PCR及Western—blot法检测肝细胞中细胞色素酶P450（CYP450）的表达。结果显示,槲皮素处理损伤的肝细胞后,肝细胞CYPIAl、CYP281及CYP2E1的表达随槲皮素浓度的增加和处理时间的延长而呈先升高后降低的趋势。10mg／LAroclor1254是诱导体外培养的原代肝细胞损伤的最适质量浓度,10μmol／L槲皮素是对损伤的怀孕大鼠肝细胞的最佳保护浓度。结果表明,槲皮素对损伤的怀孕大鼠肝细胞具有保护作用。     

Effects of some commercial preparations of polychlorinated biphenyls in growing piglets.

Sixty-four piglets were given ad libitum commercial rations containing 250 ppm of polychlorinated biphenyls as Aroclor 1232, Aroclor 1242 and Aroclor 1254 for periods up to 12 weeks. The toxic effects of the polychlorinated biphenyls upon plasma testosterone levels, the blood coagulation system, the gross- and micropathological changes as well as the extent and differences in their distribution and accumulation were studied. Principals given Aroclor 1232, Aroclor 1242 and Aroclor 1254 for the 12 week period gained 30, 10 and 50% more in body weight respectively than did controls. In the context of the present experiment, Aroclors (especially Aroclor 1254) at 250 ppm level in the ration, appeared to act as growth promoting substances. The mode and site of action of this effect is not known. The gross and micropathological examinations of controls and principals gave no evidence of anomalies. Similarly, the data on various parameters of blood coagulation did not reveal any disturbances. However, testosterone levels in plasma were elevated in male piglets given Aroclors. Analyses of polychlorinated biphenyls showed that the highest concentrations were present in adipose tissues of all treated animals. A cumulative tendency was also evident. The levels were relatively high in brain, in some instances higher than in liver of the same group. Levels in hepatic and muscular tissue were, in many instances, of the same magnitude.     

大豆黄酮对多氯联苯引起的鸡胚下丘脑星形胶质细胞损伤的缓解作用

实验建立了鸡胚下丘脑星形胶质细胞体外培养模型,研究了大豆黄酮对多氯联苯Aroclor 1254(A1254)引起的下丘脑星形胶质细胞损伤的缓解作用。结果表明:0.1～1μg/mL的A1254引起的细胞损伤不显著,高剂量时(10μg/mL)能引起星形胶质细胞大量死亡。抗氧化剂大豆黄酮10μg/mL能明显缓解A1254对星形胶质细胞的损伤作用。     

Microsomal activation of constituents of white snakeroot (Eupatorium rugosum Houtt) to form toxic products

Components of white snakeroot, a plant toxic to livestock and human beings, were activated by Aroclor 1254-induced rat liver microsomes. The toxic products of microsomal activation were evaluated in murine melanoma (B16F1) cell cultures. Toxic products in white snakeroot were inactive in cell culture systems without microsomal activation. This activation system revealed that at least 2 fractions of white snakeroot were metabolically activated to cytotoxic agents. The autocatalytic inactivator of cytochrome P-450, 1-aminobenzotriazole, inhibited activation of white snakeroot constituents by rat liver microsomes.     

Sulfoxidation of albendazole by a cytochrome P450-independent monooxygenase from rat liver microsomes

The in vitro biological oxidation of albendazole to albendazole sulfoxide by rat liver microsomes has been studied. This reaction corresponds to a NADPH-dependent enzymatic system, characterised by Km and Vm values of 53.6 microM and 0.59 nmole/mg protein per min. The rate of sulfoxidation by liver microsomes of rats treated with phenobarbital, B-naphthoflavone, Aroclor 1254 and 3-methylcholanthrene was not increased. SKF 525A and metyrapone did not inhibit albendazole sulfoxidase. Thiobenzamide and tranylcypromine decreased sulfoxidation to 48 and 52% of control values. The inhibition by tranylcypromine was competitive. Purified flavin adenine dinucleotide (FAD)-containing monooxygenase from hog liver microsomes catalysed sulfoxidation of albendazole (V = 0.52 nmole/nmole enzyme per min). The present data demonstrate that sulfoxidation of albendazole in the rat liver is not catalysed by a cytochrome P450-dependent monooxygenase and suggest that albendazole is a substrate for FAD-containing monooxygenase (FMO).     

Sulfoxidation of albendazole by a cytochrome P450-independent monooxygenase from rat liver microsomes

The in vitro biological oxidation of albendazole to albendazole sulfoxide by rat liver microsomes has been studied. This reaction corresponds to a NADPH-dependent enzymatic system, characterised by Km and Vm values of 53.6 M and 0.59 nmole/mg protein per min.The rate of sulfoxidation by liver microsomes of rats treated with phenobarbital, B-naphthoflavone, Aroclor 1254 and 3-methylcholanthrene was not increased. SKF 525A and metyrapone did not inhibit albendazole sulfoxidase.Thiobenzamide and tranylcypromine decreased sulfoxidation to 48 and 52% of control values. The inhibition by tranylcypromine was competitive. Purified flavin adenine dinucleotide (FAD)-containing monooxygenase from hog liver microsomes catalysed sulfoxidation of albendazole (V=0.52 nmole/nmole enzyme per min).The present data demonstrate that sulfoxidation of albendazole in the rat liver is not catalysed by a cytochrome P450-dependent monooxygenase and suggest that albendazole is a substrate for FAD-containing monooxygenase (FMO).     

A retrospective survey of endocardial proliferative lesions in rats

Sixty, proliferative, endocardial lesions were diagnosed in 19,304 rats, for an overall incidence of 0.3%. This population consisted of 10,127 Fischer 344, 8,737 Wistar, 200 Sprague-Dawley, and 240 Long Evans rats from chronic/oncogenicity studies reported at Lilly Research Laboratories from 1976 to 1988. Of the 60 proliferative lesions, 44 were classified as endocardial hyperplasia, 15 as endocardial schwannomas, and one as an endocardial sarcoma for prevalence rates of 0.2%, 0.08%, and 0.005%, respectively. Affected rats ranged in age from 42 to 110 weeks. There were no sex or treatment-related differences in the prevalence of the rat endocardial proliferative lesions. A review of endocardial lesions in 18 of 233 Wistar rats treated with carbamate derivatives revealed endocardial hyperplasia in 12 rats, schwannomas in five rats, and a sarcoma in one rat. One of the 12 rats with endocardial hyperplasia also had an intramural schwannoma. Of 200 Wistar rats given N-nitroso-N-methylurea, two had endocardial hyperplasia, and one had an endocardial schwannoma. Morphologic features were similar in either spontaneous or treatment-associated hyperplasia or neoplasia of the rat endocardium. Probable Schwann cell origin of the endocardial proliferative lesions was indicated by positive immunohistochemical staining for S-100 antigen in 10/12 spontaneous and 11/14 carcinogen-associated endocardial hyperplastic lesions. Further, 15/16 spontaneous and 6/7 carcinogen-associated neoplasms were immunoreactive to S-100. No tumor metastasis was recorded in either the spontaneously affected or carcinogen-treated rats.     

Granular cell lesions in the distal female reproductive tract of aged Sprague-Dawley rats

During the review of a rat carcinogenicity study, a spectrum of granular cell lesions was recognized in the distal female reproductive tract. To verify the diagnoses, cell populations of nine granular cell alterations of the cervix or vagina were characterized immunohistochemically and four were evaluated ultrastructurally. Immunoreactivity was demonstrated in 8/9 cases with S100 protein, 6/9 cases with neuron-specific enolase, and 7/9 cases with Leu-7. Granular cells were negative for smooth muscle-specific actin and calretinin. The immunohistochemical profile of these lesions was similar to that previously reported in other species, including humans. Ultrastructurally, as expected many lysosomal bodies were present in the cytoplasm of granular cells in all specimens evaluated. Based on the detailed evaluation of a series of lesions, we adopted the following diagnostic criteria and nomenclature for the granular cell changes of the female reproductive tract of rats. Granular cell aggregates were non-space-occupying lesions composed of clusters of typical granular cells. Benign granular cell tumors were space occupying lesions that typically contained prominent interstitial collagen and were either discrete masses or were difficult to discern from the surrounding tissues. Some benign tumors also contained foci of spindle cells with decreased granularity. Malignant tumors exhibited pleomorphism and an increased nucleus: cytoplasm ratio morphologically but had the same biologic behavior as the benign tumors. We applied these diagnostic criteria during the review of controls from 9 carcinogenicity studies. Up to 23% of control females in those carcinogenicity studies had granular cell lesions that could be classified into one of the three categories. Granular cell lesions appear to be common in the cervix/vagina of the Sprague-Dawley rat, and tumors may develop from granular cell aggregates.     

Hematology and clinical chemistry values in pregnant Wistar Hannover rats compared with nonmated controls

BACKGROUND: The Wistar Hannover rat has been considered as an alternative animal model to the Sprague-Dawley rat in the safety evaluation of candidate pharmaceuticals for potential reproductive and developmental toxicity. Hematology and clinical chemistry results may provide useful evidence of maternal toxicity in the absence of fetal effects. OBJECTIVE: The purpose of this study was to evaluate differences in routine laboratory values between nonmated and pregnant (near-term)Wistar Hannover rats during a control developmental study. METHODS: One hundred fifty pregnant female Wistar Hannover rats (Tac:Glx:WlfBR) were dosed orally once per day with distilled water from gestation days (GDs) 6 through 17. An additional 150 nonmated (nonpregnant) females used as age-matched controls were dosed from study days (SDs) 7 through 18. Blood samples were collected on GD 18 or 19 (SD 19 or 20) for routine hematology and plasma clinical chemistry tests. Reference intervals were established for pregnant and nonmated animals. RESULTS: On GD 18/19, pregnant rats had a lower RBC count, hemoglobin concentration, and HCT, and higher MCH, MCHC, reticulocyte percentage, and platelet, WBC, absolute reticulocyte, segmented neutrophil, lymphocyte, and monocyte counts compared with nonmated rats. Pregnant rats had lower albumin, glucose, urea, and chloride concentrations, lower creatine kinase and alkaline phosphatase activities, higher total bilirubin, cholesterol, triglyceride, calcium, phosphorus, and globulin concentrations, and higher ALT activity than nonmated rats. Serum triglyceride concentration was approximately fourfold higher in pregnant rats compared with nonmated controls. CONCLUSION: Differences in hematology and chemistry values in pregnant Wistar Hannover rats are similar to those in Sprague-Dawley rats and support use of the Wistar Hannover rat as an animal model in the assessment of maternal toxicity. Differences in laboratory values of pregnant rats should be considered when interpreting data following exposure to candidate pharmaceuticals.