共查询到20条相似文献,搜索用时 15 毫秒
1.
Intradermal challenge of Icelandic horses with extracts of four species of the genus Culicoides 总被引:1,自引:0,他引:1
Twenty-three Icelandic horses were challenged with extracts of four species of biting midges: Culicoides pulicaris, C chiopterus, C obsoletus and C impunctatus. Fourteen of the tested horses were affected with summer eczema. The horses were challenged intradermally with 0.1 ml of whole-body extracts of midges at a concentration of 0.01 or 0.005 per cent weight/volume. The skin reactions were measured after 30 minutes, 60 or 180 minutes and four, 24 and 48 hours after injection. Antigen titration showed that the reaction was dependent on the antigen concentration. Eight of nine unaffected horses failed to respond to any of the four antigens; the remaining animal responding to two of the four antigens. Ten of the 14 affected horses responded to at least three of the four antigens, while two of the animals in this group failed to respond to any. The mean responses to C chiopterus, C obsoletus and C impunctatus, read after 30 minutes, 60 minutes and four hours were significantly higher in the affected horses than in the unaffected horses. A significant difference was also found in the mean response to C chiopterus and C impunctatus, read after 24 hours. 相似文献
2.
T L Hanner J W Allen A Robertson-Byers S L Hurley 《American journal of veterinary research》1991,52(7):1065-1068
Whole-cell lysates and proteinase K-extracted lipopolysaccharide (LPS) of 19 strains of the group eugonic fermenter-4 (EF-4) were analyzed by electrophoresis and protein immunoblotting. These strains were isolated from dog- and cat-bite abscesses in human beings, ferret and human gastric lesions, and cat-lung infections. These strains represent 2 biovar groupings; EF-4a biovars ferment glucose and possess arginine dihydrolase activity, whereas EF-4b biovars do not. Electrophoresis of whole-cell lysates could distinguish between these biovars groups. Electrophoresis of LPS extracts revealed that all strains of EF-4 possess smooth chemotypes. Two strains of EF-4a reacted weekly in protein immunoblots and revealed distinct LPS profiles. These studies suggests that subgroups of EF-4 biovars may exist. 相似文献
3.
利用反转录聚合酶链式反应(RT—PCR),从猪囊尾蚴虫体中扩增出小热休克蛋白(small Heat shock protein,sHSP)基因,克隆至原核表达载体pGEX-4T-1,在大肠杆菌BL21中以GST融合蛋白的形式表达。SDS-PAGE分析证明,表达产物为64Ku的融合蛋白,经凝胶扫描分析,表达量约占菌体可溶性蛋白的30%。免疫学分析(Western blot,ELISA)结果表明,sHSP能与猪囊尾蚴血清反应。将表达产物点电泳后切胶回收免疫小鼠,进行Western blot和ELISA检测,其抗血清能与猪囊虫粗抗原及纯化的sHSP重组融合蛋白产物发生免疫学反应。该研究为丰富新的诊断试剂和免疫用重组抗原具有重要意义,同时,预测糖基化、磷酸化作用在sHSP实现生物学功能方面也具有广泛的意义。 相似文献
4.
Egenvall A Lilliehöök I Bjöersdorff A Engvall EO Karlstam E Artursson K Heldtander M Gunnarsson A 《The Veterinary record》2000,146(7):186-190
Three female beagle dogs inoculated with granulocytic Ehrlichia species were monitored for four to six months to determine whether there was evidence that the organisms persisted. The dogs were inoculated intravenously with blood containing an Ehrlichia species closely related to Ehrlichia equi and Ehrlichia phagocytophila, and identical to the human granulocytic ehrlichiosis agent with respect to its 16S rRNA gene sequence. The clinical signs were evaluated, and blood samples were collected for haematology, serum biochemistry and serology. Ehrlichial inclusions in the blood were monitored by microscopy, and ehrlichial DNA was detected by the polymerase chain reaction (PCR). Two of the dogs were injected with prednisolone on days 54 to 56 and days 152 to 154 after infection, and the other was injected with prednisolone on days 95 to 97 after infection. The dogs were euthanased and examined postmortem. Ehrlichial inclusions were demonstrated in the neutrophils and seroconversion occurred shortly after inoculation. Two of the dogs developed acute disease with rectal temperatures above 39.0 degrees C, after which no further clinical signs were observed. The administration of corticosteroids seemed to facilitate the detection of ehrlichial inclusions. Ehrlichial DNA was detected intermittently by PCR in blood samples from two of the dogs throughout the study. Persistent infection was demonstrated up to five-and-a-half months after inoculation. 相似文献
5.
Broiler minibreeder hens were used to produce monovalent antisera to bacterins prepared from serotypes 1, 3, 4, and 3 X 4 cross (CU strain) of P. multocida and to a polyvalent fowl cholera bacterin containing serotypes 1, 3, and 4. Antiserum to the CU strain (live vaccine) was also produced. Monovalent enzyme-linked immunosorbent assay (ELISA) plate antigens were prepared by separately sonicating each of the strains. Polyvalent plate antigen (Poly 3) was prepared by combining, in equal amounts after sonication, antigens from serotypes 1, 3, and 4. Each antiserum was assayed against its homologous ELISA plate antigen and against all other heterologous plate antigens, including Poly 3. The strongest reactions, as indicated by the highest absorbance values, were observed in homologous ELISAs. The CU strain may be the best monovalent ELISA plate antigen for detecting antibodies formed in response to a commercial polyvalent bacterin and to vaccinations with the live CU strain. Overall, monovalent serotype 1 (strain X-73) antiserum did not react well with any other heterologous ELISA plate antigen, whereas monovalent antisera of serotypes 4 (strain P-1662) and 3 X 4 (CU strain) reacted equally strongly with monovalent serotype 4 ELISA plate antigen. Background binding of negative serum was significantly lower (P less than 0.05) when using CU plate antigen than when using any of the other plate antigens. 相似文献
6.
低温胁迫对4种卫矛属植物抗寒生理指标的影响 总被引:1,自引:0,他引:1
《草原与草坪》2020,(2)
以一年生离体枝条为试验材料,进行人工模拟降温处理,处理温度分别为5,0,-5,-10℃,以5℃为对照,测定不同低温处理下4种卫矛属植物丙二醛(MDA)、游离脯氨酸(Pro)以及超氧化物歧化酶(SOD)、过氧化物酶(POD)活性等4项指标,并运用隶属函数法对其抗寒性进行综合评价。结果表明:低温胁迫下,4种卫矛属植物枝叶内的MDA和Pro含量,SOD和POD活性,均显著高于对照(5℃)(P<0.05);4种卫矛属植物抗寒性强弱的排序为:无柄卫矛>刺果卫矛>胶州卫矛>扶芳藤。 相似文献
7.
Chia-Chia Huang Yu-Chen Hsieh Chau-Loong Tsang Yang-Tsung Chung 《Journal of veterinary science (Suw?n-si, Korea)》2010,11(4):333-340
Ehrlichia (E.) canis is a Gram-negative obligate intracellular bacterium responsible for canine monocytic ehrlichiosis. Currently, the genetic diversity of E. canis strains worldwide is poorly defined. In the present study, sequence analysis of the nearly full-length 16S rDNA (1,620 bp) and the complete coding region (4,269 bp) of the gp200 gene, which encodes the largest major immunoreactive protein in E. canis, from 17 Taiwanese samples was conducted. The resultant 16S rDNA sequences were found to be identical to each other and have very high homology (99.4~100%) with previously reported E. canis sequences. Additionally, phylogenetic analysis of gp200 demonstrated that the E. canis Taiwanese genotype was genetically distinct from other reported isolates obtained from the United States, Brazil, and Israel, and that it formed a separate clade. Remarkable variations unique to the Taiwanese genotype were found throughout the deduced amino acid sequence of gp200, including 15 substitutions occurring in two of five known species-specific epitopes. The gp200 amino acid sequences of the Taiwanese genotype bore 94.4~94.6 identities with those of the isolates from the United States and Brazil, and 93.7% homology with that of the Israeli isolate. Taken together, these results suggest that the Taiwanese genotype represents a novel strain of E. canis that has not yet been characterized. 相似文献
8.
9.
The antigenic interrelatedness of 3 clone-purified turkey reoviruses (NG-Turkey, 82-88, and NC-TEV) to each other and to 4 clone-purified chicken reoviruses (S1133, Co8, Fahey-Crawley, and avian type 2) was determined in reciprocal cross-neutralization tests, using polyclonal antisera and the plaque-reduction technique. The morphologic features of plaques formed under agar were studied for all 7 reoviruses, and size comparisons for turkey vs chicken isolates were made. All 3 turkey reoviruses (with the exception of NG-Turkey vs Fahey-Crawley chicken reovirus) formed plaques significantly (P less than 0.05) smaller than plaques produced by their chicken counterparts. The 3 turkey reoviruses were closely related to each other and to chicken reovirus CO8. The antigenic differences between turkey reoviruses 82-88 and NC-TEV and chicken reovirus S1133 were slight (minor subtype); however, the latter and NG-Turkey were serotypically distinct. The NG-Turkey and 82-88 turkey reoviruses were more related (minor subtype) to the Fahey-Crawley and avian type 2 chicken reoviruses, than was NC-TEV turkey reovirus (major subtype). 相似文献
10.
11.
The detection of Anaplasma and Ehrlichia species is usually based on species-specific PCR assays, since no assay is yet available which can detect and identify these species simultaneously. To this end, we developed a reverse line blot (RLB) assay for simultaneous detection and identification of Anaplasma and Ehrlichia species in domestic ruminants and ticks. In a PCR the hypervariable V1 region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for members of the genera Anaplasma and Ehrlichia [Int. J. Syst. Evol. Microbiol. 51 (2001) 2145]. Amplified PCR products from blood of domestic ruminants or Amblyomma variegatum tick samples were hybridized onto a membrane to which eight species-specific oligonucleotide probes and one Ehrlichia and Anaplasma catch-all oligonucleotide probe were covalently linked. No DNA was amplified from uninfected blood, nor from other hemoparasites such as Theileria annulata, or Babesia bigemina. The species-specific probes did not cross-react with DNA amplified from other species. E. ruminantium, A. ovis and another Ehrlichia were identified by RLB in blood samples collected from small ruminants in Mozambique. Finally, A. variegatum ticks were tested after feeding on E. ruminantium infected sheep. E. ruminantium could be detected in adult ticks even if feeding of nymphs was carried out 3.5 years post-infection. In conclusion, the developed species-specific oligonucleotide probes used in an RLB assay can simultaneously detect and identify several Ehrlichia and Anaplasma species. However, as no quantitative data for the detection limit are available yet, only positive results are interpretable at this stage. 相似文献
12.
Matsumoto K Takeuchi T Yokoyama N Katagiri Y Ooshiro M Zakimi S Gaowa Kawamori F Ohashi N Inokuma H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2011,73(11):1485-1488
We collected a total of 206 Haemaphysalis longicornis ticks by flagging in pastures in Yonaguni Island, Okinawa, Japan, in April 2008. Four of the 206 tick DNA samples tested were positive in a polymerase chain reaction (PCR) screening for the 16SrRNA gene of Anaplasmataceae. Partial sequences of 4 PCR products were identical to each other. Longer sequences of the 16SrRNA gene were successfully determined in 2 of the 4 tick samples, and the obtained 1,392 bp and 1,300 bp sequences revealed high similarity to the 16SrRNA gene sequences of the validated Ehrlichia species, including Ehrlichia ewingii, E. chaffeensis, and E. canis (98.3-98.6%). We also sequenced 1,304 bp of the groEL gene from the 2 tick samples, and found that these had the highest similarity to sequences of E. ewingii (94.0-94.4%) in the validated ehrlichial species. Based on the 16SrRNA and groEL gene sequences, the ehrlichial agents detected in this study were similar to the Ehrlichia species detected in Asia and may compose a new Ehrlichia species with other Ehrlichia species detected in Asia. 相似文献
13.
Quantitative immunoelectrophoretic methods have been used to study the antigenic mosaic of Peptococcus indolicus, an anaerobic coccus frequently isolated from udder secretions from heifers and dry cows with mastitis. Three antigenic components of liquid cultures of this bacterium were analyzed, compared and characterized, namely concentrated culture filtrate containing extracellular antigens, a cytoplasmic antigen fraction obtained by freeze-press disruption of bacterial cells and Triton X-100-soluble antigens from cell wall-membrane fractions. The extracellular antigens were further investigated because they proved to be particularly useful in preliminary studies on the antibody response of cows to P. indolicus. The possible cross-reactivity of peptococcal antigens with extracellular antigens from other bacteria causing, or associated with, mastitis was investigated. The contribution of medium components to the immunoprecipitate profile, the heat-stability of antigens and the relationship of serotypic antigens to those in the standard extracellular concentrate were established using co-immunoelectrophoresis, crossed-line immuno-electrophoresis and crossed immunoelectrophoresis with intermediate gel. Attempts to identify enzyme-active immunoprecipitates with histochemical enzyme staining methods revealed only glutamate dehydrogenase. 相似文献
14.
Antigenic analysis of avian Chlamydia psittaci using monoclonal antibodies to the major outer membrane protein. 总被引:1,自引:0,他引:1
A Kikuta N Furukawa T Yoshida H Fukushi T Yamaguchi K Hirai 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1991,53(3):385-389
Monoclonal antibodies to the major outer membrane protein (MOMP) of Chlamydia psittaci derived from a parrot were established for antigenic analysis of avian C. psittaci. With 17 monoclonal antibodies to MOMP, 17 reactivity patterns were identified on 112 strains of C. psittaci, C. pneumoniae and C. trachomatis, which were isolated from birds, mammals and humans in Japan, U.S.A., Canada and Taiwan, from 1938 to 1987. Immunological reactivity of budgerigar-derived strains to the monoclonal antibodies was different from that of pigeon-derived strains. Imported bird-derived strains were distinguishable from domestic bird-derived strains by the reactivity to the monoclonal antibodies. A close relationship between the subtypes and geographic origins was indicated on budgerigar-derived strains. On the contrary, various reactivity patterns were shown in pigeon-derived strains isolated in a narrow area. The monoclonal antibodies established in the present work may be useful probes for ecological study of avian C. psittaci. 相似文献
15.
16.
Watanabe M Oikawa T Hiraoka H Kaneko N Itamoto K Mizuno T Okuda M Inokuma H 《Veterinary parasitology》2006,136(2):147-154
Three beagle dogs were inoculated with mice spleen/liver homogenate infected with Ehrlichia species detected from Ixodes ovatus (EIO) and one dog was used as a control. All three infected dogs did not show clinical signs of disease except for mild pyrexia throughout the 41-day study period. Splenomegaly was observed from Day 7 post-inoculation (p.i.) in two of the dogs. Hematological and biochemical abnormalities included mild thrombocytopenia, hypoproteinaemia, hypoalbuminaemia and increased C-reactive protein values. One of the dogs' splenic aspirate sample was PCR-positive for Ehrlichia Day 7 p.i. and another dogs' blood and bone marrow aspirate sample was PCR-positive Day 41 p.i. Sequence analysis of the PCR products showed 100% homology with the 16SrRNA partial gene sequence of Ehrlichia sp. HF565. Antibody titers to EIO were observed in all three experimentally infected dogs starting from the first week p.i. and cross-reactivity with Ehrlichia canis was detectable in one of the dogs starting Day 7 p.i. These data suggest that infection of dogs with EIO is possible, though is probably of low pathogenic importance. Cross-reactivity of EIO infected dog serum with E. canis raises the likelihood of false E. canis seropositive dogs. 相似文献
17.
Antigenic variation of Mycoplasma gallisepticum, as detected by use of monoclonal antibodies. 总被引:8,自引:0,他引:8
V S Panangala M A Morsy M M Gresham M Toivio-Kinnucan 《American journal of veterinary research》1992,53(7):1139-1144
A panel of monoclonal antibodies (MAb) developed against Mycoplasma gallisepticum strain PG31 was used to probe the antigenic profiles of 5 recognized strains (PG31, R, S6, F, A5969) and 6 field isolates of M gallisepticum. Monoclonal antibody G9 predominantly recognized antigens at apparent molecular mass positions of 90 to 98 kDA. The MAb reacted with all strains and isolates, but the molecular mass position of the antigens varied among some mycoplasmas. Monoclonal antibody G12 reacted with all strains and isolates of M gallisepticum and had an identical banding pattern. However, MAb G10 and G11 reacted selectively only with a limited number of strains and/or isolates. Surface distribution of the MAb-recognized antigens was revealed by immunoelectron microscopy. Partial physicochemical characterization of MAb G9-recognized antigens identified glycopeptide characteristics. Monoclonal antibody G9 reacted with surface antigens and, hence, participated in agglutination of M gallisepticum. However, the degree of agglutination varied among the various strains and isolates, indicating a quantitative or conformational limitation or an alteration in the anomeric expression of the epitopes. Antigenic variation in M gallisepticum may be mediated by immunologic selective pressures, or a proclivity for habit niche in the host. 相似文献
18.
19.
Antigenic analysis of Pasteurella haemolytica serovars 1 through 15 by crossed immunoelectrophoresis
Pasteurella haemolytica serovars 1 through 12, grown in broth and on agar plates, and 2 field isolates (types A1 and T10) were used to develop polyvalent crossed immunoelectrophoresis (XIE) reference systems. The maximal number of antigens was revealed by XIE when sonicates of agar plate-grown organisms were used as the immunogen (to produce antibodies) and as the soluble antigen for XIE. Antigens produced from agar plate-grown organisms were less contaminated (by antigenic components of the medium) than were those produced from organisms grown in broth. Seventy-two antigens were detected in sonicated preparations of agar plate-grown P haemolytica. The common antigen of gram-negative bacteria was identified in the P haemolytica XIE reference system; precipitation was observed with rabbit antiserum to the common antigen of gram-negative bacteria isolated from Escherichia coli, as well as with rabbit immunoglobulins (obtained from unvaccinated rabbits). Most preimmune sera from our vaccinated rabbits also precipitated the common antigen. Serovar-specific antigens in the P haemolytica XIE reference system were defined and presumptively identified as part of the bacterial lipopolysaccharide complex by use of the limulus amebocyte lysate test. Partial cross-reactions were found between serovar-specific antigens within each biovar (A and T). Pasteurella haemolytica biovar A-specific and biovar T-specific antigens were defined by crossed-line immunoelectrophoresis. When serovars A13, A14, and T15 were tested in the P haemolytica XIE reference system, they gave high matching coefficient values of 0.98, 0.98, and 0.87, respectively. The proposal to separate P haemolytica biovars A and T into 2 different species was supported by immunotaxonomic data obtained from crossed immunoelectrophoresis, but more extensive studies will be necessary to establish the appropriate taxonomic position of these 2 groups of organisms. 相似文献
20.
Peixoto CC Marcelino I Vachiéry N Bensaid A Martinez D Carrondo MJ Alves PM 《Veterinary microbiology》2005,107(3-4):273-278
Ehrlichia ruminantium (ER) is the causative agent of Heartwater, one of the most common tick-borne diseases affecting ruminants in African countries and West Indies. Although ER can be used as an inactivated vaccine for wild and domestic animals, there are currently no easy and reliable methods for the quantification of this obligate intracellular bacterium. This report describes the development of a SYBR Green I based real time PCR protocol for the quantification of ER for vaccine production purposes. The method was validated for four ER strains. The external-standard-based PCR protocol developed has a large dynamic quantitative range allowing accurate ER measurement in samples containing from 10(2) to 10(8) gene copies; the method is also reproducible and precise, with intra- and inter-assay coefficients below 5%. The detection limits were validated for samples collected from bovine aortic endothelial cell culture bulks, which are commonly used to produce the ER vaccine. In contrast to the methods based upon protein content, no interference from the host cells in ER quantification was observed. Furthermore, the extended applicability of the new technique was demonstrated by monitoring ER production in cell culture thus rendering it a valuable tool to ensure consistency between vaccine lots and to evaluate optimal vaccine dosage. 相似文献