绵羊瘤胃微生物胞外蛋白酶性质的研究

本试验测定了pH、温度和EDTA对纯化的绵羊的瘤胃微生物胞外蛋白酶活性的影响,并用一已知氨基酸排列顺序的肽段(由15个氨基酸组成)作为底物,进一步测定了蛋白酶的酶切位点。结果表明,(1)纯化蛋白酶的最适pH在6.0～6.5之间,最适温度为40℃左右。(2)不同浓度的EDTA(浓度分别为1,10,25,50,75和100mM)对蛋白酶活性没有影响,因而该酶属于丝氨酸蛋白酶类。(3)纯化蛋白酶是一种内肽酶,其酶切位点在组氨酸—酪氨酸(?)、天冬氨酸—丙氨酸(?)、亮氨酸—赖氨酸(?)和(或)缬氨酸—赖氨酸(?)相连的肽键。(4)胰蛋白酶和纯化的蛋白酶虽同属丝氨酸蛋白酶类,但前者对底物具有高度的专一性,后者对底物的专一性不强,这是瘤胃内饲料蛋白质强烈降解的主要原因之     

Pathogenicity of porcine enterotoxigenic Escherichia coli that do not express K88, K99, F41, or 987P adhesins.

Three-week-old weaned and colostrum-deprived neonatal (less than 1 day old) pigs were inoculated to determine the pathogenicity of 2 enterotoxigenic Escherichia coli isolates that do not express K88, K99, F41, or 987P adhesins (strains 2134 and 2171). Strains 2134 and 2171 were isolated from pigs that had diarrhea after weaning attributable to enterotoxigenic E coli infection. We found that both strains of E coli adhered in the ileum and caused diarrhea in pigs of both age groups. In control experiments, adherent bacteria were not seen in the ileum of pigs less than 1 day old or 3 weeks old that were noninoculated or inoculated with a nonpathogenic strain of E coli. These control pigs did not develop diarrhea. Antisera raised against strains 2134 and 2171 and absorbed with the autologous strain, grown at 18 C, were used for bacterial-agglutination and colony-immunoblot assays. Both absorbed antisera reacted with strains 2134 and 2171, but not with strains that express K99, F41, or 987P adhesins. A cross-reaction was observed with 2 wild-type K88 strains, but not with a K12 strain that expresses K88 pili. Indirect immunofluorescence with these absorbed antisera revealed adherent bacteria in frozen sections of ileum from pigs infected with either strain. We concluded that these strains are pathogenic and express a common surface antigen that may be a novel adhesin in E coli strains that cause diarrhea in weaned pigs.     

Estrogen and progesterone receptor status of mammary carcinomas and correlation with clinical outcome in dogs.

Estrogen and progesterone receptors (ER, PR) were measured in cytosol fractions from 18 primary canine mammary carcinomas by use of biochemical assays. One or both receptors were detected (> 10 fmol/mg of cytosol protein) in 11 tumors: 5 ER and PR; 2 ER only; 4 PR only. Mean cytoplasmic receptor concentrations (fmol/mg of cytosol protein) were 22.8 +/- 2.9 (SEM) for ER and 51.0 +/- 10.3 for PR in tumors containing ER and PR, 28.8 +/- 12.1 for ER in tumors containing only ER and 13.2 +/- 1.5 for PR in tumors containing only PR. Estrogen or progesterone receptors or both were identified in 6 of 9 tubular adenocarcinomas, 4 of 5 papillary adenocarcinomas, and 1 of 1 squamous cell carcinoma. These receptors were not identified in solid carcinomas (n = 2) or a single spindle cell carcinoma. Although the number of cases was limited, survival times of dogs tended to be longest in those with tumors containing ER alone or in combination with PR, intermediate in those with tumors containing only PR, and shortest in those with tumors without ER or PR. A correlation was not apparent between receptor status and age, presence of ovaries, tumor size, or histologic classification of the tumor. In the analysis of this series, the extent of surgery (mastectomy of the involved gland vs unilateral or bilateral mastectomy) did not appear to influence the outcome of the disease, and metastasis to regional lymph nodes did not appear to be a reliable prognostic indicator.     

Rhinoscopy in dogs and cats]

Rostral and caudal rhinoscopy in dogs and cats facilitates the investigation of the nasal cavity and accurate biopsy. Rostral rhinoscopy can be performed by rigid endoscopes; caudal rhinoscopy requires flexible endoscopes. Deep anaesthesia or additional analgesia with local anaesthesia is necessary. The nasal cavity is assessed by its form, colour, surface of the mucous membrane, hyperemia, plaques, lesions, and the secretion is assessed by its quantity, colour and viscosity. Foreign bodies and neoplasia must also be looked for. Case reports with abnormal findings are described.     

Infection of bovine fetuses at 120 days' gestation with virulent and avirulent strains of bluetongue virus serotype 11.

Bluetongue virus infection in sheep and cattle during fetal development causes neuropathology. Two strains of bluetongue virus serotype 11 designated as UC-2 and UC-8 have different virulence patterns in newborn mice. These viruses have distinctly different electropherotype patterns on polyacrylamide gel electrophoresis indicating a genetic difference in these two viruses of the same serotype. Four bovine fetuses each were inoculated intramuscularly with either UC-2 or UC-8, and one fetus was inoculated with placebo. The inoculation was made intramuscularly through the uterine wall at 120 days' gestation, and the bovine fetuses were recovered by cesarean section 12 or 20 days after inoculation. Fetal blood was collected for virus isolation and serology. Virus was reisolated from brain, blood, lung and liver. Both strains, UC-2 and UC-8, cause severe lesions in the 120 day fetuses. The encephalomalacic lesions occurred earlier and were more severe in fetuses inoculated with UC-8 as compared to those inoculated with UC-2. The subtle differences observed in the fetuses inoculated with the two different strains suggest that there is a difference in pathogenic potential of the two viruses. These differences do not appear to be completely dependent upon the host species.     

Feline immunodeficiency virus neurotropism: evidence that astrocytes and microglia are the primary target cells.

To investigate the neuropathogenesis of feline immunodeficiency virus (FIV) infection in vitro, we have utilized three populations of cultured feline neural cells (astrocytes, microglia, brain endothelium) to assess the relative susceptibility to FIV infection, ability to produce viral antigens, and effects of infection on cell survival. Astrocytes appeared to be the most susceptible to infection, followed by microglia, whereas brain endothelial cells were relatively resistant to infection. Astrocyte infection resulted in syncytium formation and cell death, while microglial cells remained persistently and productively infected, without obvious cytopathic effects. These results suggest that FIV entry into the central nervous system probably does not occur via infected endothelium and that both astrocytes and microglia are more likely target cells for the virus.     

大肠杆菌耐热性肠毒素ST1—K99／LacZ基因重组的研究

将构建的ｐＢＳＴ２～６工程菌质粒用限制性内切酶ＢａｍＨＩ／ＢｇｌⅡ酶切，经琼脂糖凝胶电泳和电洗脱，回收１４７ｂｐ的目的ＳＴ１基因。随之将该基因分别重组到能有效表达Ｋ９９菌毛抗原和ＬａｃＺ酶的ｐＧＫ９９之Ｋ９９基因ＢｇｌⅡ位点和ｐＵＣ１８的ＢａｍＨＩ位点中。通过ＳＴ１基因探针菌落原位杂交、特定酶切分析及ＤＮＡ序列分析，筛选并鉴定出了理想重组子，从而构建出了能分别表达ＳＴ１融合基因产物的工程菌株ｐＳＫ２１９和ｐＸＳＴ１。     

Relationship of common avian pathogen antibody titers in so-called chicken anemia agent (CAA)-antibody-positive chicks to titers in CAA-antibody-negative chicks.

Antibody titers for infectious bursal disease virus (IBDV), infectious bronchitis virus, Newcastle disease virus, and reovirus from chicks with chicken anemia agent (CAA) antibodies were compared with antibody titers from their CAA-antibody-negative counterparts. These comparisons were made in 396 chickens that were 1 day, 2 weeks, 8-9 weeks, 10 weeks, 17 weeks, or 29-32 weeks old. Only one serum sample was collected from any given chick or chicken. There were no significant differences between the antibody titers at any age for any antigen, with one exception: at 29-32 weeks, the IBDV titers were higher (t = 2.62, df = 142, P less than 0.01) in chickens with CAA antibody. Although not at all likely, we believe that the observation of high IBDV antibody titers in CAA-antibody-positive chicks could have been a spurious one.     

Biopsy of the bovine mammary gland.

A technique is described for biopsy of the bovine udder, employing sedation and local anaesthesia. Tissue samples of approximately 5 g were obtained by electrocautery from two quarters of the udder of a cow laterally recumbent. Care was taken to ensure complete haemostasis which was achieved by electrocoagulation and ligation. Postoperative recovery was rapid, and loss of yield was no greater in biopsied glands than in control glands of the same cow. Yield from all quarters returned to preoperative levels within 48 h.     

Monoclonal antibodies to bovine viral diarrhea virus: cross-reactivities to field isolates and hog cholera virus strains.

Monoclonal antibodies to bovine viral diarrhea virus (BVDV) were examined for binding with a large number of North American BVDV isolates and eight strains of the serologically related pestivirus, hog cholera virus (HCV). No single BVDV monoclonal antibody reacted with all BVDV isolates. The most cross-reactive monoclonal antibody was an anti-p80/p125 antibody which showed a positive reaction with 173 of 180 (96%) North American isolates. From a fewer number of isolates tested, one anti-gp53 monoclonal antibody also showed a high cross-reactivity (94%). All BVDV isolates showed a positive reaction with at least one of the seven monoclonal antibodies in the panel. Thus, the results indicated that a pool of these monoclonal antibodies may be used in place of polyclonal antisera for the detection of BVDV contamination of cell lines or for virus isolation. For HCV, all three anti-p80/p125 monoclonal antibodies reacted positively with all eight virus strains. In contrast, none of the anti-gp53 monoclonal antibodies were reactive to HCV strains. Thus, the anti-gp53 monoclonal antibodies may be useful for distinguishing between usually innocuous BVDV infections and the highly significant HCV infections in swine for foreign animal disease surveillance.