首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 328 毫秒
1.
Mycoplasma hyopneumoniae causes pneumonia in pigs. The effect of infection by this organism on histochemical characteristics of airway mucin within epithelial cells was studied. Seven- to 10-week-old pigs were inoculated intratracheally with M hyopneumoniae or culture broth, and lung tissues were collected from inoculated and control pigs at 2, 4, and 6 weeks after inoculation. Tissue sections were stained with periodic acid-Schiff/Alcian blue, pH 2.5 or high iron diamine/Alcian blue. Histologic features of randomly selected bronchi, bronchioles, and submucosal glands were compared in sections stained with periodic acid-Schiff/Alcian blue. Bronchial goblet cell sulfomucin and sialomucin were quantitated by image analysis of sections stained with high iron diamine/Alcian blue. Bronchi and bronchioles of infected pigs contained proportionately fewer goblet cells with mucin at all stages of infection than age-matched control pigs. Goblet cells in bronchi of infected pigs contained significantly less total mucin and sialomucin, and significantly more sulfomucin than goblet cells of control pigs. Increased sulfated mucin in bronchial goblet cells may reflect altered glycoprotein production or secretion in response to infection with M hyopneumoniae.  相似文献   

2.
Pigs aged 6 to 9 weeks from enzootic pneumonia-free herds were inoculated intranasally with a suspension of pneumonic lung containing Mycoplasma hyopneumoniae or were placed in contact with such inoculated pigs. All the inoculated pigs had gross lesions of enzootic pneumonia when killed 27 to 42 days after inoculation. The culture methods described enabled M. hyopneumoniae to be isolated from all 29 inoculated pigs. Of 45 pigs in contact with inoculated pigs 35 had gross lesions of enzootic pneumonia when killed 28 to 71 days later and M. hyopneumoniae was isolated from 33. Another 9 had lesions, detected only microscopically, and M. hyopneumoniae was recovered from 3 of these when killed 75 to 98 days after contact began. In a separate experiment M. hyopneumoniae isolated from experimentally infected pigs, and adapted to the culture medium after 6 passages, caused gross lesions of enzootic pneumonia in 1 of 4 pigs inoculated intranasally.  相似文献   

3.
The 6-chloro analogue of norfloxacin (compound A) administered continuously in the feed at 400 ppm for 21 days markedly reduced the extent and activity of pneumonic lesions in pigs with pneumonia induced experimentally with an homogenate of pneumonic lung and broth cultures of Mycoplasma hyopneumoniae. Norfloxacin at 100 ppm or compound A at 200 ppm in the feed did not reduce the extent of lung lesions, although half the pigs treated with norfloxacin had lesions which appeared histologically to be healing. M hyopneumoniae was detected either by culture or immunofluorescence in the lungs of 60 per cent of the pigs treated with compound A at 400 ppm compared with all the pigs in the other groups. These results were related to the amount of drug in the lungs and body fluids during therapy. Only compound A at 400 ppm produced concentrations in the lungs and bronchial secretions exceeding the minimum inhibitory concentration against M hyopneumoniae. Mycoplasmacidal concentrations were not reached either in the lungs or bronchial secretions which might account partly for the frequent detection of M hyopneumoniae in the lungs after treatment. Drug resistance did not appear to be responsible for the persistence of M hyopneumoniae in vivo since the M hyopneumoniae isolates from the pigs after therapy were sensitive in vitro to both quinolones. As daily weight gain and feed-conversion efficiency improved in all groups of treated pigs compared with the controls, these effects were probably unrelated to the antimycoplasmal activities of the two quinolones.  相似文献   

4.
Lungs from 191 slaughter pigs with gross lesions indicative of enzootic pneumonia of pigs (EPP) and 80 grossly normal lungs, all originating from 9 different herds, were subjected to microbiological and pathological examinations. The microbiological studies included both bacterial and mycoplasmal culture and also testing for Mycoplasma hyopneumoniae antigen in tissue by indirect immunofluorescent technique. M. hyopneumoniae, Pasteurella multocida and Mycoplasma hyorhinis were detected in 83%, 43% and 37% of the pneumonic lungs, respectively. Mycoplasma flocculare was the most frequently isolated organism in the non-pneumonic lungs. The greatest amounts of macroscopic pneumonia (25.2%) were recorded in lungs with all the three agents M. hyopneumoniae, P. multocida and M. hyorhinis present. The amounts of pneumonia in lungs with M. hyopneumoniae alone and in concurrence with P. multocida, were 9.3% and 15.6%, respectively. M. hyorhinis was also, in this study, associated with higher frequency of diffuse pleuritis. These findings indicate that M. hyorhinis might be involved in the pathogenesis of pneumonia in slaughter pigs. Ninety-six per cent of the isolates of P. multocida from pneumonic lungs could be characterized as type A. In the herds which had the most severe pneumonia problems, toxin production was detected in 83% of the P. multocida strains while only 28% were toxigenic in herds with subclinical to moderate pneumonia problems.  相似文献   

5.
Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia, a chronic nonfatal disease affecting pigs of all ages. To obtain better insight in the mechanisms responsible for differences in virulence between highly and low virulent M. hyopneumoniae isolates, 23 caesarean-derived, colostrum-deprived piglets were randomly assigned to three groups. Groups 1 and 2 consisted of nine animals each, which were intratracheally inoculated at 1 week of age with a highly or a low virulent isolate of M. hyopneumoniae, respectively. The remaining five animals were inoculated with sterile culture medium. Animals were euthanized at 5, 10, 15 and 28 days post-inoculation (DPI). Animals inoculated with the highly virulent isolate had more neutrophils in BAL fluid at 10, 15 and 28DPI compared to the other groups. At 10 and 15DPI, animals in the highly virulent group had significantly higher concentrations of TNF-alpha in BAL fluid. IL-1beta concentration in this group was higher at 5 and 28DPI compared to the other groups. From 10DPI onwards, significantly higher titres of M. hyopneumoniae were detected in the BAL fluid of animals inoculated with the highly virulent isolate compared to animals inoculated with the low virulent isolate. Additionally, the in vitro generation time of the highly virulent M. hyopneumoniae isolate was significantly shorter than that of the low virulent isolate. The present study indicates that the difference in pathogenicity between the highly and low virulent isolates is associated with a faster in vitro growth, a higher capacity to multiply in the lungs and the induction of a more severe inflammation process by the highly virulent isolate.  相似文献   

6.
The interaction between Mycoplasma hyopneumoniae and Pasteurella multocida in experimental pneumonia was investigated in conventional pigs. The experimental animals were 49 days old when inoculated with M. hyopneumoniae; they were inoculated with P. multocida after 23 days, and killed 13 days later. In pigs inoculated only with P. multocida, clinical signs and lung lesions were not observed, and the agent was not recovered. Pigs inoculated with M. hyopneumoniae developed fever, moderate cough and dyspnea which tended to disappear, and small proliferative lung lesions from which M. hyopneumoniae was isolated. Pigs inoculated with both agents had higher fever, severe cough and dyspnea which tended to aggravate, and extensive exudative lung lesions from which organisms were isolated. All animals had similar growth rates, but the group infected with both agents consumed 60% more food. Therefore, M. hyopneumoniae causes mild pneumonia, whereas P. multocida is not pathogenic alone but aggravates the pneumonia initiated by M. hyopneumoniae.  相似文献   

7.
Cellular and humoral immune responses of pigs inoculated with Mycoplasma hyopneumoniae were investigated at postinoculation weeks (PIW) 2, 4, and 6. The response of blood lymphocytes (BL) and bronchial lymph node lymphocytes (LNL) to stimulation by M hyopneumoniae antigens was evaluated by a lymphocyte-stimulation test. Specific antibodies in serum and lung washing samples were assayed by ELISA. Immunoglobulin-positive cells in lungs and bronchial lymph nodes were identified by indirect fluorescent antibody test, using isotype-specific monoclonal antibodies. At PIW 0 to 6, BL from control and M hyopneumoniae-inoculated pigs were stimulated by M hyopneumoniae cells; however, BL from inoculated pigs generally had higher stimulation indices, especially at PIW 6. The response of LNL was influenced by previous exposure to M hyopneumoniae, as indicated by higher stimulation indices (P less than 0.01) of LNL from inoculated pigs killed at PIW 2 and 6. Specific ELISA antibodies to M hyopneumoniae in lung washings from inoculated pigs consisted mainly of IgG and IgA isotypes. Examination of lung sections by indirect immunofluorescence revealed that cells producing IgM and IgA were in controls as well as M hyopneumoniae-inoculated pigs, but IgG-positive cells were only in lungs of inoculated pigs. Resolution of pneumonia appeared to correlate with development of increased sensitization of BL, as well as development of marked increases in immunoglobulins, particularly IgG in lung washings at PIW 6.  相似文献   

8.
To evaluate the transmission of Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus (PRRSV) by aerosol as either a single or mixed infection, 28 pigs were inoculated intratracheally with M hyopneumoniae on day 0 and infected intranasally with PRRSV on day 35; they were housed together in a barn. To assess the aerosol transmission of M hyopneumoniae as a single infection, one trailer (A) containing 10 five-week-old sentinel pigs was placed along the south side of the infected barn (1 m from the fans) on day 28. To assess the mixed infection, two trailers (B and C), each containing 10 five-week-old sentinel pigs, were placed along each side of the barn on day 42. The sentinel pigs in the three trailers were exposed to the exhaust from the fans for seven days. No M hyopneumoniae infection was detected in the sentinel pigs in trailer A, but it was detected in the sentinel pigs in trailers B and C. No PRRSV was detected in any of the sentinel pigs.  相似文献   

9.
The effect of immunosuppression on Mycoplasma hyopneumoniae infection was evaluated by comparing data from infected, thymectomized, and antithymocyte serum-treated pigs (group 1) with data from infected (group 2) and noninfected (group 3) healthy pigs. After groups 1 and 2 pigs were inoculated intranasally with M hyopneumoniae, mycoplasmas tended to multiply slightly more in the lungs and bronchial lymph nodes of group 1 pigs than that of group 2 pigs. Organisms were also isolated from the spleen of 1 of 3 group 1 pigs. Pneumonia developed in group 2 pigs and was characterized by massive peribronchial, peribronchiolar, and perivascular lymphoid hyperplasia and exudate consisting mainly of polymorphonuclear leukocytes in the alveoli and lumina of the bronchioles and bronchi. In group 1 pigs, perivascular and peribronchiolar cuffings by lymphocytes were less prominent, and the extent of intraluminal exudate was severe and widespread. Bronchial lymph nodes from group 2 pigs had marked hyperplasia of germinal centers and paracortical areas. In group 1 pigs, germinal centers were hyperplastic, whereas in the paracortical areas, depletion of lymphocytes was evident. Seemingly, cell-mediated immune mechanisms are important in the development of pneumonic lesions in enzootic pneumonia of pigs.  相似文献   

10.
Microorganisms associated with pneumonia in slaughter weight swine.   总被引:2,自引:1,他引:1       下载免费PDF全文
The lungs of 334 pigs were obtained from two slaughter plants in Minnesota and examined in detail. Macroscopic and microscopic evaluation, direct fluorescence for Mycoplasma hyopneumoniae and bacterial culture were done on all of them and a subsample of 50 were selected for virus culture. Mycoplasma hyopneumoniae, Pasteurella multocida and Haemophilus spp. were detected in 24.0%, 34.1% and 27.0% of the lungs, commonly in conjunction with each other. One isolate of Haemophilus pleuropneumoniae serotype 2 was detected and this represents the first report of its presence in the United States. No virus was detected in any of the lungs. Lungs with both M. hyopneumoniae and Pasteurella multocida had the greatest amount of macroscopic pneumonia (9.8% of the lung). Lungs with M. hyopneumoniae or P. multocida alone had 4.9% and 5.2% of the lung involved with pneumonia respectively. Lungs with Haemophilus sp. Taxon "minor group" had 3.8% of the lung involved which was not significantly different from lungs with none of these organisms being detected (1.6%). There was a positive correlation between the extent of M. hyopneumoniae infection, as scored by FAT and the amount of macroscopic pneumonia present (r = 0.46; P less than 0.001). Likewise, there was a positive correlation between the estimated concentration of P. multocida present, as scored by the relative number of colonies on blood agar and the amount of macroscopic pneumonia present (r = 0.60; P less than 0.001). Microscopically, the amount of lymphoreticular proliferation, polymorphonuclear cells and alveolar macrophages were evaluated.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

11.
The efficacy of in-feed medication with tylosin for the treatment of enzootic pneumonia was examined in an experimental Mycoplasma hyopneumoniae infection model. One group of 10 conventional M. hyopneumoniae-free pigs was inoculated intratracheally with a highly virulent field isolate of M. hyopneumoniae; a second group of 10 pigs was inoculated in the same way and after 12 days was given tylosin at 100 mg/kg feed for 21 days; a third group of 10 pigs was inoculated with sterile culture medium, and these pigs were not given tylosin. The pigs were examined daily for clinical signs and each pig was given a respiratory disease score. Thirty-three days after they had been infected the pigs were euthanased, the lung lesions were quantified and samples of lung were processed for immunofluorescence testing for M. hyopneumoniae. The mean (sd) respiratory disease and lung lesion scores were significantly higher (P<0.05) in both the infected groups than in the uninfected group. Between 23 and 33 days after infection the mean respiratory disease score of the pigs treated with tylosin was 0.54 (0.22), significantly (P<0.05) lower than that of the infected pigs which were left untreated, 1.54 (0.46); similarly, their average lung lesion score, 1.72 (1.20), was significantly lower than that of the untreated pigs, 5.27 (3.85).  相似文献   

12.
Fano E  Pijoan C  Dee S 《The Veterinary record》2007,161(15):515-520
Twenty-eight 10-week-old pigs were inoculated intratracheally with 1 x 10(5) colour-changing units/ml Mycoplasma hyopneumoniae strain 232, and another 32 pigs were not inoculated but were divided into 12 direct-contact pigs and 20 indirect-contact pigs. Thirty-five days later, the inoculated pigs were inoculated intranasally with 1 x 10(2.4) tcid50 of porcine reproductive and respiratory syndrome virus (PRRSV) strain mn 30-100. Viraemia, seroconversion and the transmission of PRRSV in the M hyopneumoniae-infected pigs were then assessed for four months. Three groups of 10 age-matched gilts were introduced as sentinels into the experimental barn on days 28, 56 and 84 after the PRRSV infection. The persistence of PRRSV was evaluated in both the experimentally and naturally infected pigs, which were slaughtered 120, 135 and 150 days after the infection. The period of viraemia and the extent of seroconversion were similar to those observed in studies of pigs infected only with PRRSV, suggesting that under the conditions of the study M hyopneumoniae did not affect these features of the disease. A delayed pattern in the seroconversion and proportion of pcr-positive pigs was observed in the direct and indirect contact groups, and the persistence of PRRSV in tissues was confirmed by pcr at 120 and 150 days after infection only in the directly inoculated pigs and not in the direct- or indirect-contact groups of pigs.  相似文献   

13.
The virulence of a laboratory adapted culture of Mycoplasma hyopneumoniae strain NB12 was determined in three- to five-day-old gnotobiotic piglets. Intranasal inoculation or exposure to an aerosol of the culture caused low incidences of pneumonia in the piglets. Passage of M hyopneumoniae strain NB12 in gnotobiotic piglets resulted in a rapid increase in virulence. After only three in vivo passages, severe pneumonia involving most lobes of the lung developed in all inoculated piglets within three and a half weeks. All 49 piglets inoculated with the piglet-passaged NB12 strain in nine subsequent experiments developed pneumonia but the extent of the pneumonic lesions varied considerably from piglet to piglet. The histopathology of the lung lesions was similar to that reported as being induced by other strains of M hyopneumoniae in gnotobiotic piglets and resembled that seen previously in conventionally reared neonatal piglets inoculated with homogenised lung from pigs with enzootic pneumonia. Aspiration pneumonia caused by milk inhalation occurred in some piglets. The pneumonia induced with the piglet-passaged NB12 strain was judged to be suitable for the study of porcine enzootic pneumonia or for the evaluation of chemotherapeutic agents.  相似文献   

14.
The effect of dextran sulfate (DS), known to be cytotoxic to macrophages, on the cell-mediated and humoral immune response to nonviable Mycoplasma hyopneumoniae in pigs was investigated. The cell-mediated immune response was determined by means of lymphocyte transformation a test, using uptake of [3H]thymidine in a microculture system and the humoral immune response by means of a microplate complement-fixation test. Peripheral blood lymphocytes from pigs vaccinated with nonviable M hyopneumoniae and DS incorporated substantially more [3H]thymidine than did those from pigs given Mycoplasma or DS alone. The transformation of lymphocytes from M hyopneumoniae-DS vaccinated pigs was enhanced when M hyopneumoniae cells used in the assay system were heated at 60 C for 30 minutes. Similarly prepared M flocculare and M hyorhinis cells also stimulated lymphocytes from M hyopneumoniae-DS vaccinated pigs, but not nearly as great as when M hyopneumoniae cells were used. The humoral antibody response and the cell-mediated immune response to nonviable M hyopneumoniae was markedly enhanced by DS. Pigs were vaccinated with nonviable M hyopneumoniae and/or DS 4 times and challenge exposed intratracheally with viable M hyopneumoniae. Pigs vaccinated with M hyopneumoniae and DS had less severe pneumonia than did nonvaccinated pigs.  相似文献   

15.
Groups of six male goats were inoculated intratracheally and intranasally with either caprine herpesvirus followed 6 days later by Pasteurella haemolytica, canine herpesvirus alone or P. haemolytica alone. Pneumonic lesions were observed in five of the six goats inoculated with caprine herpesvirus followed by P. haemolytica and in three of the six goats inoculated with P. haemolytica alone, but were not observed in goats inoculated with caprine herpesvirus alone or in non-infected controls. Pasteurella haemolytica was isolated from seven of eight lungs with pneumonia, but only from one of sixteen lungs without pneumonia. The lesions ranged from fatal acute exudative necrotising pneumonia to predominantly proliferative pneumonia. Half of the caprine herpesvirus-inoculated goats developed a clinical catarrhal rhinitis five days post-inoculation and the only virus-specific histopathological lesion was a mild tracheitis. Canine herpesvirus was recovered from the nasal swabs of all caprine herpesvirus- inoculated goats developed a clinical catarrhal rhinitis five days post-inoculation and the only virus-specific histopathological lesion was a mild tracheitis. Canine herpesvirus was recovered from the nasal swabs of all canine herpesvirus-inoculated goats and from the lungs of three goats inoculated with caprine herpesvirus alone. The experimental inoculations demonstrated that P. haemolytica alone can produce pneumonia in goats. In addition, the study showed that caprine herpesvirus readily proliferates in the upper respiratory tract and lungs of goats but the role of caprine herpesvirus in the aetiology of pneumonia remains uncertain.  相似文献   

16.
Enzootic pneumonia (EP) of pigs, caused by Mycoplasma hyopneumoniae has been a notifiable disease in Switzerland since May 2003. The diagnosis of EP has been based on multiple methods, including clinical, bacteriological and epidemiological findings as well as pathological examination of lungs (mosaic diagnosis). With the recent development of a real-time PCR (rtPCR) assay with 2 target sequences a new detection method for M. hyopneumoniae became available. This assay was tested for its applicability to nasal swab material from live animals. Pigs from 74 herds (average 10 pigs per herd) were tested. Using the mosaic diagnosis, 22 herds were classified as EP positive and 52 as EP negative. From the 730 collected swab samples we were able to demonstrate that the rtPCR test was 100% specific. In cases of cough the sensitivity on herd level of the rtPCR is 100%. On single animal level and in herds without cough the sensitivity was lower. In such cases, only a positive result would be proof for an infection with M. hyopneumoniae. Our study shows that the rtPCR on nasal swabs from live pigs allows a fast and accurate diagnosis in cases of suspected EP.  相似文献   

17.
Arginine-utilising mycoplasmas isolated from the pneumonic lungs of 4 pigs were identified as M. hyosynoviae by their growth characteristics, biochemical activity and serological similarity to each other and to 2 type strains of M. hyosynoviae in growth inhibition tests. A purified culture of one M. hyosynoviae isolate was inoculated intravenously into 3 pigs which were killed 9 days later. Although arthritis was not observed clinically, M. hyosynoviae was isolated from 8 of 9 joints cultured, the tonsils and retropharyngeal lymph nodes of all 3 pigs and the lungs of 2 pigs. No histopathological changes were observed in synovial membranes of the joints cultured or lungs but severe depletion of lymphocytes in the cortex of lymphoid follicles in lymph nodes and tonsils was observed.  相似文献   

18.
To gain further insight into the pathogenesis of porcine enzootic pneumonia (PEP), cytokine expression in different pulmonary compartments was examined. Mycoplasma hyopneumoniae (Mh) and proinflammatory and immunoregulatory cytokines (IL-1alpha, IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10 and TNF-alpha) were detected by immunohistochemical methods in porcine lungs experimentally infected with Mh. Ten pigs were inoculated intranasally with Mh and killed in pairs weekly from 1- to 5-week post-inoculation (wpi). Three Mh-free pigs were taken as controls. Mh-antigen was shown in paraffin-wax-embedded tissues by immunohistochemistry in the luminal surface of bronchial and bronchiolar epithelial cells of all Mh-infected pigs. Significant increase in cytokine expression was detected on snap-frozen tissues from the bronchoalveolar exudate of the airways, mononuclear cells of the alveolar septa and macrophages and lymphocytes of the peribronchial and peribronchiolar lymphoid tissue, from 1 wpi onwards, compared to expression in non-pneumonic lungs. The main cytokines in the BALT of Mh-infected animals that showed an increase were IL-2, IL-4, IL-8, IL-10 and TNF-alpha. In the alveolar septa and bronchoalveolar exudate IL-1 (alpha and beta), IL-2, IL-4, IL-8 and IL-10 expression also increased in infected animals.  相似文献   

19.
Effect of enzootic pneumonia of pigs on growth performance   总被引:8,自引:0,他引:8  
Growing pigs were naturally infected with a field strain of Mycoplasma hyopneumoniae to assess the effect of enzootic pneumonia on production. Both the initial ("breakdown") and endemic stages of infection were evaluated. The pigs were reared under environmental and management conditions commonly found on commercial piggeries in South Australia. Growth rate of pigs held in-contact with inoculated pigs was reduced by 12.7% (p less than 0.01) between 50 to 85 kg bodyweight. In the second trial inoculated gilts were used to naturally infect piglets during suckling. Growth rate of infected pigs was reduced by 15.9% (p less than 0.001) between 8 to 85 kg bodyweight, while feed conversion was depressed by 13.8% (p less than 0.05) between 10 to 25 kg bodyweight. At current feed and production costs this reduced performance added approximately $2.80 to the cost of every pig produced. These losses were recorded in groups of pigs in which enzootic pneumonia was present. At slaughter, 40% of lungs contained gross lesions of enzootic pneumonia which were free of significant secondary bacteria. The nature of the infection was established by gross and microscopic pathology and confirmed by the detection of specific complement fixing antibody in infected pigs and the demonstration of M. hyopneumoniae by direct immunofluorescent staining of lung sections.  相似文献   

20.
Immunofluorescent staining has been used to identify Mycoplasma hyopneumoniae in smears of broth cultures, in infected pig testicle cell cultures, and in frozen cut sections of pneumonic lungs from field and experimentally produced cases of enzootic pneumonia. In the pneumonic pig lung, fluorescent staining was limited to the surface of the bronchial and bronchiolar epithelium and to the contained exudate. In a series of trials using experimentally infected pigs fluorescence was not detected until 25 days post-infection and was regularly seen in pigs killed thereafter. Porcine immune globulin precipitated from the serum of experimentally infected pigs and conjugated with fluorescein isothiocyanate was reactive and specific for the detection of M. hyopneumoniae. Immune globulin conjugates prepared from the serum of hyperimmunized rabbits were reactive but in some cases produced a faint non-specific staining of frozen tissue sections. No such non-specific reactions were noted on stained culture smears or cell cultures.

Fluorescence was not seen in known positive preparations stained with non-immune pig globulin conjugates or in preparations from uninoculated cell cultures or pigs, stained with non-immune or immune globulin conjugates.

Mycoplasma hyorhinis was detected by immunofluorescent staining with homologous conjugates, in smears of broth cultures and in tissue sections from pigs with polyserositis.

Immunofluorescent staining was found to be species specific and useful for the early species identification of mycoplasma isolated from pigs.

  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号