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1.
D F Guo H Kodama M Onuma T Kimura M Yoshimizu 《The Japanese journal of veterinary research》1991,39(1):27-37
Seven strains of Oncorhynchus masou virus (OMV) genomes were analyzed with the restriction endonucleases BamHI, EcoRI, HindIII and SmaI. The restriction patterns of OMV strain DNAs were divided into four groups. Restriction profiles of high passage strains (00-7812, 65th passage, and H-83, 60th passage) were different from those of low passage strains (00-7812, 8th passage, and H-83, 6th passage) when digested with BamHI, HindIII and SmaI. However, no difference was observed between the restriction patterns of high and low passage viral DNA with EcoRI. There was no distinct difference observed between the restriction patterns of tumor tissue-derived and coelomic fluid-derived strains. By using 32P-labelled DNA of standard OMV (strain 00-7812) as a probe, most of the fragments of other OMV strain DNAs were hybridized. 相似文献
2.
J Varga 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1991,38(7):497-504
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were used to identify and to compare the surface antigens of eight C. fetus subsp. fetus strains. Seven strains (one of serogroup A and six of serogroup B) were isolated from aborted ovine fetuses, while one strain (serogroup A) originated from an aborted calf fetus. Saline extracts at 56 degrees C and 100 degrees C were used as antigens. Antisera were produced in rabbits. In saline extracts (56 degrees C) of the strains at least 19 fractions were identified by SDS-PAGE, with molecular masses ranging from approx. 4,800 to 205,000. The major bands appeared at 205,000, 66,000, 31,500, 25,000, 21,000 and 17,500. Despite the fact that the strains were cultured from 4 different sheep flocks and belonged to serogroup A or B, the SDS-PAGE profiles of the strains were very similar. When boiled (100 degrees C) extracts were used, a band migrating at 32,500 in sheep strains and a band at 97,500 in the calf isolate were missing. Most of the bands obtained by SDS-PAGE could be identified also by the immunoblot procedure. A or B type specificity of the ovine isolates was due to an LPS fraction, migrating at approx. 21,000, while the other LPS fractions appearing under this region although reacted with antisera did not influence the type specificity. Using alkaline extracts (pH 12) in SDS-PAGE, LPS fractions gave more pronounced profiles. In two of our C. fetus subsp. fetus isolates, plasmids with a molecular mass of 31,500 were identified. 相似文献
3.
A 39 kDa protein mediates adhesion of avian Pasteurella multocida to chicken embryo fibroblast cells 总被引：5，自引：0，他引：5
Borrathybay E Sawada T Kataoka Y Ohtsu N Takagi M Nakamura S Kawamoto E 《Veterinary microbiology》2003,97(3-4):229-243
To clarify the role of avian Pasteurella multocida capsule in pathogenesis, adhesion of capsulated strains P-1059, X-73 and Pm-18, and noncapsulated strains P-1059B, Pm-1 and Pm-3 to chicken embryo fibroblast (CEF) cells was compared. Number of adherent organisms of the capsulated strains to CEF cells were approximately three times as much as noncapsulated strains indicating that adhesive properties were enhanced by the presence of bacterial capsule. Pretreatments of the bacterial cells with heat, trypsin, or with antiserum caused a marked decrease in adhesion of capsulated strain P-1059 and its noncapsulated variant P-1059B. However, depolymerization of capsular hyaluronic acid with high dose of hyaluronidase enhanced adhesion of these strains. Combined treatments of the bacterial cells with both hyaluronidase and trypsin significantly (P < 0.05) inhibited the adherence of strain P-1059 as compared to the treatment only with trypsin, but strain P-1059B was not affected. SDS-PAGE profiles of crude capsular extract (CCE) prepared from capsulated strain P-1059 and its noncapsulated variant P-1059B grown on dextrose starch agar (DSA) plates by heating at 56 degrees C in a 2.5% NaCl solution demonstrated eight protein bands of 28, 34, 36, 39, 52, 56, 63 and 93 kDa. The 28, 34 and 36 kDa proteins were commonly major for both strains, and the 39 kDa protein was major only for strain P-1059 but poor in strain P-1059B. Outer membrane protein (OMP) profiles were identical with a major protein at 34 kDa and four minor proteins between the two strains. The adhesion of strain P-1059 and strain P-1059B to CEF cells was inhibited significantly (P < 0.01) by treatment with rabbit antisera against P-1059, P-1059B, CCE or 39 kDa protein of strain P-1059 as compared to the treatment with either PBS or with normal rabbit serum. These results indicated that an antigenic 39 kDa protein in the capsule may be responsible for adhesion of avian P. multocida type A strains to CEF cells as a virulence factor. 相似文献
4.
2004年在病猪体内分离到一株猪链球菌,通过平板扩散法和微量稀释法药敏试验表明这株链球菌对红霉素敏感。采用这株猪链球菌进行体外诱导试验,在低浓度药物组第165代和高浓度药物组第180代的菌液对红霉素M IC值均达到中介水平。它们的耐药表型均为内在型,而且都扩增到了ermB耐药基因。其中180代菌的23S rRNA碱基1387位A突变成G;它们的核糖体蛋白L4,165代菌碱基104位、585位和633位,分别T突变成C、A突变成G和A突变成G,180代菌碱基283位和651位,分别A突变成G和T突变成G;核糖体蛋白L22,165代和180代菌碱基109位和468位,分别C突变成A和T突变成A,并且165代菌碱基还在426位G突变成A。这些碱基的突变可能是引起猪链球菌对红霉素耐药的原因之一。 相似文献
5.
A Mycoplasma gallisepticum strain designated 6/85 (MGI) exhibiting reduced virulence for both chickens and turkeys was sequentially passaged 10 times in each species. DNA extracted from organisms before passage and those isolated after the third, sixth, and 10th passages was studied by restriction endonuclease DNA analysis using BamHI, BglII, EcoRI, HindIII, and PstI endonucleases. The virulent-type strain designated S6 was used as a comparison. Comparison of DNA fragment patterns of MGI and S6 strains showed distinct differences, although some similarities were evident. Passage of the strain in vivo did not affect DNA fragment patterns of the MGI strain. Electrophoretic protein patterns produced by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed very similar band patterns in both the MGI and S6 strains. The most notable differences were seen in bands located in the molecular-mass regions of approximately 46.5, 50-54, 58-64, and 105-140 kilodaltons. Alteration of band pattern profiles following in vivo passage of the MGI strain was apparent in a single band at approximately 86 kilodaltons that appeared to stain more intensely following passage. 相似文献
6.
A molecular analysis of strains of Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and Mycoplasma mycoides subsp. mycoides, small colony type (M. mycoides SC) isolated from goats was performed using the amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE) fingerprinting techniques. Among the 11 field strains of M. capripneumoniae from Tanzanian goats, two AFLP patterns were demonstrated, with 10 of the strains showing indistinguishable patterns. Five Kenyan strains of M. capripneumoniae produced three AFLP patterns, with two of them being indistinguishable from the 10 identical Tanzanian and one Ugandan strain (M74/93) isolated from sheep. The AFLP pattern of the type strain (F38(T)) was identical to two Kenyan strains (Baringo and G183/82). On PFGE analysis, all the examined M. capripneumoniae strains exhibited identical PFGE profiles.Five field strains of M. mycoides SC isolated from goats displayed identical AFLP patterns except for one strain which differed from others at only one position. The AFLP pattern of the type strain of M. mycoides SC (PG1(T)) was different from the field strains. The five field strains of M. mycoides SC produced identical PFGE profiles, which were, however, different from the type strain. The AFLP and PFGE profiles of M. mycoides SC strains from goats were identical to those of six strains isolated from cattle affected with contagious bovine pleuropneumonia (CBPP) in the same areas. The results of this study suggest a close epidemiological linkage between strains of M. capripneumoniae and between M. mycoides SC type, respectively, isolated from goats in Tanzania. 相似文献
7.
A A Adesiyun A Viebahn H G Sahl W Lenz K P Schaal 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1992,39(1):39-47
The lytic activity, protein profile and morphology of five newly isolated phages from canine Staphylococcus aureus strains and one from a human S. aureus strain were compared with those of selected phages in the international phage sets (IPS). Five canine phages lysed 57 (76.0%) of 75 canine isolates of Staphylococcus aureus from Nigeria at routine test dilution (RTD) while 34 (IPS) phages typed only 31 (41.3%) strains at RTD or/and 100-RTD. The new human phage lysed 11 (14.7%) of 75 strains isolated from human diarrhoea. The new phages were readily propagated, specific in activity and stable during storage at 4 degrees C. Prominent proteins detected by SDS-PAGE indicated similarities between some of the phages but one canine phage was distinctly different, as was its morphology which was an isometric head with a short tail compared to oval heads and long tails which characterized others. IPS phages in the same serologic group had similar protein profiles but no correlation was observed with lytic groups. The use of protein profile and electron micrographs allowed classification of the phages into serogroups. It is concluded that the newly isolated canine phages could be very useful in typing Nigerian canine strains of S. aureus. 相似文献
8.
Field strains of Aujeszky's disease virus (ADV) were attenuated by heat treatment and serial passage at sub-optimal growth temperatures in chicken embryo fibroblasts (CEF). At chosen passage levels, virus was titrated in cell culture and in mice. For each strain, the pathogenicity was expressed as a mouse lethal index (MLI), defined as the inverse of the log10 (CCID50:LD50). MLIs determined for field strains displayed a wide range of comparatively high values. The attenuation of field strains was accompanied by a rapid fall in MLI values, particularly in the initial stages. Heat-treated ADV attenuated faster than untreated ADV, when passaged at 30 degrees C. Passage at 27 degrees C resulted in considerably accelerated attenuation compared to passage at 30 degrees C, in the case of both untreated and heat-treated ADV. MLIs were determined for attenuated ADV strains that had been tested in 6-day-old piglets. Low MLI values were found to correlate with low virulence in piglets and high MLI values with virulence. 相似文献
9.
Cytotoxins in non-enterotoxigenic strains of Escherichia coli isolated from feces of diarrheic calves 总被引：10，自引：0，他引：10
We have examined the cytotoxic responses produced in HeLa and Vero cell cultures by sonicates from 15 non-enterotoxigenic (STa-, LT-) strains of E. coli, highly lethal for mice parenterally LD50 less than 3 X 10(7) CFU), which had been isolated from feces of diarrheic calves. Three types of cytotoxic responses were observed. Type 1 (five strains) consisted of enlargement, rounding and polynucleation of HeLa cells, an effect previously reported with cytotoxic necrotizing factor (CNF) in E. coli from infant and piglet enteritis. Type 2 toxicity (three strains and the control Vir strain S5) was also characterized by enlargement and polynucleation of HeLa cells, but in contrast to Type 2 effect, cells were elongated. Sonicates from the latter strains were lethal for chickens, producing the lesions previously described with Vir strains. Type 3 toxicity (two strains and the control VT strain H19), produced an extensive destruction of both Vero and HeLa cell cultures. Cytotoxic effects were completely abolished upon heating for 1 h at 60 degrees C for Type 1 and 2 extracts and at 80 degrees C for Type 3 extracts. Seroneutralization assays showed that cytotoxins of the same type were closely related antigenically. In addition, a slight cross-neutralization was observed between Type 1 (CNF) and Type 2 (Vir) toxins. 相似文献
10.
2型猪链球菌强毒株与非致病株胞外蛋白的比较蛋白质组初步研究 总被引：1，自引：0，他引：1
为建立2型猪链球菌(S.suis 2)胞外蛋白组双向电泳样品的制备方法,本研究利用双向电泳(2-DE):分离S.suis 2强毒株与无致病株的胞外蛋白,分别得到它们的胞外蛋白图谱.在pH4～pH7范围内采用考马斯亮蓝R350染色法在2菌株的胞外蛋白质图谱中都分别检测出180±10个蛋白点.并且它们的蛋白质分子质量分布基本相似;在所检测到差异蛋白点中,其中有50个蛋白点只存在于无致病菌株中而强毒株中不存在,有52个蛋白点只存在于强毒株中而无致病菌株中不存在,有7个蛋白点在2菌株中的表达量相差5倍以上.我们在强毒株凝胶中挑取了10个差异蛋白点进行质谱分析,通过数据库检索,鉴定了其中的9个蛋白,另外1种与鞭毛蛋白有同源序列.这些结果为研究S.suis2的致病机理提供了蛋白质组学方面的信息. 相似文献
11.
Seventy per cent of the United Kingdom isolates of Salmonella typhimurium from calves are phage type 204c and the study of the epidemiology of this organism requires additional methods of strain characterisation. This paper describes the applications of biotyping and plasmid-profile analysis for this purpose. One hundred and eleven isolates from 73 outbreaks of disease were examined. All belonged to the same primary biotype, although strains from 39 of the outbreaks differed in secondary tests in failing to ferment m-inositol at 25 degrees C. Four different antibiotic resistance patterns were detected among the isolates, which possessed seven distinct plasmid profiles. The spread of a distinct type through the calf marketing chain was investigated by using these techniques. 相似文献
12.
13.
Hydrogen peroxide (H2O2) production and oxygen uptake during the oxidation of NADH and L-alpha-glycerophosphate (GP) by lysed cells was determined for the type and field strains of Mycoplasma bovis and M. agalactiae. NADH oxidation by all the strains showed variable production of H2O2 ranging from 0 to 1.21 mol/mol O2 taken up. All strains were unable to oxidize GP, showing absence of GP oxidase activity. Some strains were identified that produced relatively high levels of H2O2 (> 1.0 mol/ mol O2 taken up). In vitro passage of M. bovis strain 119B96 showed reduced H2O2 production: 0.52, 0.16, and 0.07 mol/mol O2 taken up after the 50th, 100th and 200th passages, respectively. SDS-PAGE analysis showed the loss of a protein band of 32 kDa after 50 passages. These preliminary studies show that not only does H2O2 production by potentially pathogenic Mycoplasma spp. vary in the field but also that similar alterations can be induced by passage in culture. In the latter case, at least in one M. bovis strain, this alteration has been shown by SDS-PAGE to be associated with a loss of specific protein production. Further study of these phenomena is essential background for the production of more efficient vaccines for mycoplasmas. 相似文献
14.
Outer membrane protein profiles of Yersinia ruckeri 总被引：1，自引：0，他引：1
R L Davies 《Veterinary microbiology》1991,26(1-2):125-140
The outer membrane protein (OMP) profiles of 135 isolates of Yersinia ruckeri, obtained from nine European countries (100 isolates), North America (23 isolates), Australia (six isolates) and South Africa (two isolates), and including four reference strains, were examined by SDS-PAGE. Outer membranes were isolated by selective solubilisation of the cytoplasmic membrane with 0.5% (w/v) sodium N-lauroyl sarcosinate (Sarkosyl). Outer membrane proteins were stable after in vitro passage and there was no variation in OMP profiles due to colony selection. With the exception of a 39.5 kDa peptidoglycan-associated protein there was also no variation at different stages of the growth cycle. The 39.5 kDa protein was not produced during logarithmic growth phase but increased in abundance as the stationary phase progressed. Interstrain variation occurred in the possession of a 36.5 or 38 kDa heat-modifiable protein and in the possession of peptidoglycan-associated proteins in the molecular weight range 36.5 to 40.5 kDa. Based on variation of these proteins five OMP-types, designated OMP-types 1-5, were identified among the 135 isolates examined. Outer membrane protein analysis was demonstrated to be useful in epidemiological studies of Y. ruckeri. 相似文献
15.
Bernáth S Német L Tóth K Morovján G 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2001,48(1):73-79
Protein profiles of six Erysipelothrix rhusiopathiae strains, five Erysipelothrix tonsillarum strains and three Erysipelothrix strains of uncertain taxonomic position were studied by sodium dodecyl sulphate-polyactylamide gel electrophoresis (SDS-PAGE). In a computerized comparison of the protein patterns of the strains, the level of similarity between the strains was determined. The SDS-PAGE protein bands were divided into 14 groups based on molecular weight. The relative distribution of proteins within these groups was used to characterize the strains. These distribution patterns were analysed by computing Pearson's correlation coefficient between strains, and by cluster analysis based on Euclidean distances and the unweighted pair-group method of arithmetic averages (UPGMA). The geometric mean of the similarities calculated by Pearson's correlation coefficient was 0.980 +/- 0.018 between the E. rhusiopathiae strains and 0.979 +/- 0.013 for E. tonsillarum strains. The value was 0.932 +/- 0.036 between the strains belonging to different species. However, a threshold value applicable for identification of a given strain to a species could not be established. Of the three strains of uncertain taxonomic position, the strains designated Rotzunge and Iszap 4 had a protein composition more similar to that of E. tonsillarum than to that of the E. rhusiopathiae type strain. The strain designated Pécs 56, which may be a member of a new species according to literature data, gave inconsistent results by the two methods used. The computerized evaluation method developed here is suitable for the comparison of the protein composition of the strains and for the construction of the protein similarity tree by cluster analysis. 相似文献
16.
Werner JA Kasten RW Feng S Sykes JE Hodzic E Salemi MR Barthold SW Chomel BB 《Veterinary microbiology》2007,122(3-4):290-297
The influence of in vitro passage on Bartonella henselae pathogenesis in cats has not been thoroughly evaluated. Our objective was to examine the bacterial kinetics and humoral immune responses in cats experimentally infected with three different in vitro passages of B. henselae F1, a genotype I strain of feline origin. The F1 strain was in vitro passaged 20 and 40 times, and each was inoculated into a group of 5 cats. The kinetics of bacteremia and the feline humoral immune response to bacterial antigens were compared to a previous study involving a group of six cats inoculated with the original F1 strain. Among the three groups of cats, the kinetics of bacteremia profiles and the humoral immune responses to B. henselae lysates were similar. The influence of passage on bacterial membrane proteins was examined. In vitro passage altered the expression of 4/17 (23.5%) bacterial membrane proteins and 6/15 (40%) bacterial membrane antigens. An association between poor seroreactivity to three lysate antigens (15-, 18- and 45kDa), prolonged bacteremia and decreased serum bactericidal activity was noted. Our data show that in vitro passage of B. henselae did not alter the kinetics of bacteremia, including the occurrence of relapsing bacteremia, in experimentally infected cats. This suggests that highly passaged strains may not be suitable for future vaccination studies. Furthermore, in vitro passage results in phenotypic and antigenic changes in the bacterial membrane protein profile, which warrants caution in the interpretation of studies involving passaged B. henselae strains. 相似文献
17.
A study of the heterogeneity of isolates of Mycoplasma ovipneumoniae from sheep in New Zealand. 总被引：1，自引：0，他引：1
To investigate the heterogeneity of Mycoplasma ovipneumoniae, sixty isolates from three sheep on each of twenty farms were examined by restriction endonuclease analysis (REA) and SDS-PAGE. All were found to be different except for three isolates obtained from one farm. The protein and REA patterns of individual isolates were both highly reproducible and remained unchanged following long term passage (approximately 400 generations) in vitro. No plasmids were detected in the twelve strains which were examined and when two isolates were co-cultured in vitro, no genetic interchange, as judged by changes in REA patterns were detected. Since the heterogeneity of M. ovipneumoniae when examined by SDS-PAGE is too great to allow groups to be recognised, it could be advantageous for this purpose if only surface proteins were compared. As a preliminary step to this end we have identified several surface proteins of M. ovipneumoniae and found that some are common to all strains, one surface protein was shared by five of the eight strains examined and another was unique to one strain. This approach has the potential to allow the recognition of grouping of M. ovipneumoniae isolates. 相似文献
18.
The survival period of 22 S. gallinarum strains in chicken feces was examined. The suspension of the bacterium was homogenized with a certain feces quantity. The initial ratio was 10(7) to 10(9) colony forming units per gram of feces. The results show that the growth of 5 strains was completely inhibited within 24 hours post homogenization, 7 strains were still positive to S. gallinarum for 24 hours, 5 strains were positive for 48 hours and the last 5 strains were positive for 4 days. Additionally, the effect of tryptose soy (TSB) and Rappaport-Vassiliadis (RV) nutrient broths on the isolation rate of S. gallinarum from feces was examined at 37 degrees C and 43 degrees C. It was shown that the TSB medium was the best at 37 degrees C, in this experiment. The S. gallinarum concentration in RV medium was decreased at 37 degrees C from 9.1 x 10(8) to 1.6 x 10(6) and at 43 degrees C from 9.1 x 10(8) to 4.1 x 10(2). 相似文献
19.
C.J. Thorns M.G. Sojka I.M. McLaren M. Dibb-Fuller 《Research in veterinary science》1992,53(3):300-308
A panel of 13 monoclonal antibodies from different hybridomas was produced against a novel salmonella fimbrial antigen expressed predominantly by Salmonella enteritidis strains. The specificity of the monoclonal antibodies to this antigen (SEF14) was confirmed by enzyme-linked immunosorbent assay (ELISA) using purified SEF14, immune electron microscopy and, with 11 monoclonal antibodies, the identification of a repeating protein subunit (14,300kDa) on the antigen. Blocking-ELISA with the monoclonal antibodies identified epitopes in at least three, non-overlapping clusters which appeared evenly distributed on SEF14 in immune electron microscopy. The use of the monoclonal antibodies in direct-binding ELISA on a range of salmonella serotypes suggested that the epitopes on SEF14 are highly conserved and were expressed by all the S enteritidis strains examined; some strains of S dublin and the only strain of S moscow available were the only other serotypes that expressed SEF14. A latex agglutination reagent based on a monoclonal antibody was developed and used to test for SEF14 on 280 strains (representing 120 serotypes in 24 serogroups of salmonellae) that had been grown on Sensitest agar for 18 hours at 37 degrees C. All S enteritidis strains (64) and most S dublin strains (28 of 33) produced SEF14 as did the two strains representing S blegdam and S moscow. SEF14 was not detected in any other strains of serotypes from serogroup D or from any other serogroup examined. 相似文献
20.
对鸡新城疫病毒(NDV)B1株、La Sota株、Mukteswar株和长春强毒野毒株(C87E7)感染的鸡胚尿囊液分别进行浓缩和提纯,并作SDS-PAGE和Western印迹,进行结构蛋白及其抗原性的分析。结果,4株NDV的电泳图谱显示11-12条结构蛋白带,分子量从43000到120000不等。其中各株均有3条主要蛋白带,8～9条次要蛋白带。不同株NDV的次要蛋白带有明显的区别。Schiff氏试剂染色证明,分子量为76000、52000和50000的蛋白为糖蛋白。结构蛋白的抗原性分析表明,4株NDV蛋白带中有2条具有共同抗原性,其他蛋白带的抗原性则有差异或明显不同。 相似文献