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1.
Faecal swabs obtained from 126 calves and 118 cows in Argentina were investigated for the presence of verotoxin-producing Escherichia coli (VTEC). VTEC strains were recovered from 10 (23%) of 43 calves with diarrhoea, from 24 (29%) of 83 healthy calves, from 40 (44%) of 91 healthy cows waiting at the slaughterhouse, and from 6 (22%) of 27 healthy grazing cattle. PCR showed that 21 (9%) of animals carried VT1+ strains, 49 (20%) VT2+ strains and 10 (4%) VT1+ VT2+ strains. VT1+ strains predominated among calves (16% versus 0.8%; p < 0.001). The presence of eae gene was significantly more frequent among VTEC strains isolated from calves (78%; 46/59) than from cows (2%; 1/65) (p < 0.001). Furthermore, eae gene was more prevalent in VT1+ strains (97%; 32/33) than in VT2+ strains (14%; 10/70) (p < 0.001) and in VT1+ VT2+ strains (24%; 5/21) (p < 0.001). Sorbitol negative high virulent strains serogroups O157 were not detected. This study indicates that cattle are a reservoir of VTEC strains, and that eae gene is associated with VT1+ strains that are predominating among young animals. Fortunately, only adult animals are taken to the slaughterhouse, among which VTEC strains negative for eae gen are predominating.  相似文献   

2.
Fifty verocytotoxin (VT)-producing Escherichia coli (VTEC) strains of serogroup O157 were characterized by phage typing, polymerase chain reaction (PCR) for VT genes and the E. coli attaching and effacing (eae) gene, and random amplified polymorphic DNA-PCR (RAPD-PCR) fingerprinting. The collection represented isolates obtained from patients with diarrhoea-associated haemolytic-uraemic syndrome (D+ HUS) and their family contacts, isolated in the Netherlands, Belgium and Germany between 1989 and 1993. Based on isolates from separate families (n = 27) seven different phage types were identified, types 2 (44%) and 4 (33%) were predominant. Eighty-five percent of the strains contained only VT2 gene sequences and 15% both VT1 and VT2. All strains of the dominant phage types 2 and 4 carried the VT2 gene. Strains that belonged to the minor phage types 8, 14, 32 carried both VT1 and VT2 genes, with the exception of two isolates identified as phage types 49 and 54 which contained only VT2 genes. All O157 VTEC strains possessed the chromosomally-located eae gene, which indicates its usefulness as virulence marker. RAPD-PCR fingerprinting identified four distinct banding patterns, with one profile found among 79% of the strains. Based on the combined results of all typing methods used in this study, the collection of 50 O157 VTEC strains could be divided into nine distinct groups. Strains isolated from different persons within one family could not be distinguished by any of these methods. The data suggest that O157 VTEC strains are members of one clone that has become widely distributed.  相似文献   

3.
The distribution of the Escherichia coli attaching and effacing (eae) gene in strains of verotoxin-producing E. coli (VTEC) isolated from cattle and humans was studied. The majority of strains isolated from humans with bloody diarrhoea or HUS and cattle with severe diarrhoea were eae positive (82 and 83% respectively). In contrast, 59% of VTEC isolated from asymptomatic cattle were eae negative and of the remaining 41% that were eae positive, the majority were serotype O157. H7. The nucleotide sequence of the 3'' end of the eae gene of enteropathogenic E. coli (EPEC) of serotype O55. H7 was found to be almost identical to that of serotype O157. H7. Specific primers are described which detect the eae sequences of VTEC serotypes O157. H7, O157. H-, and EPEC serotypes O55. H7 and O55. H-. The nucleotide sequence of the 3'' end of the eae gene of serotype O111. H8 differed significantly from that of O157. H7. Primers were developed to specifically identify the eae sequences of VTEC serotypes O111. H- and O111. H8. We conclude that whereas the majority of VTEC associated with disease in cattle and humans possess the eae gene, the gene itself may not be necessary to produce haemorrhagic colitis and HUS. Sequence heterogeneity in the 3'' end of eae alleles of VTEC permits specific identification of subsets of these organisms.  相似文献   

4.
Twenty verocytotoxigenic Escherichia coli (VTEC) O118 strains isolated between 1996 and 1998 from human patients in Germany were analysed for their serotypes, their virulence markers and their epidemiological relatedness. Three strains were typed as O118:H12, these carried only the VT2d-Ount variant gene and were not associated with diarrhoea or haemolytic uraemic syndrome (HUS). Seventeen strains were serotyped as O118:H16 or O118:non-motile (NM). These carried all the genes for VTI, eae and EHEC-haemolysin. The O118:H16/NM strains were from diarrhoea (13 cases) and HUS (2 cases). Sixteen of the patients were young infants and most infections were associated with a rural environment. Evidence for zoonotic transmission from cattle to humans was found in two cases. The epidemiological relationship between the human and bovine O118:H16/NM isolates was indicated by homogeneous plasmid patterns and by very similar XbaI restriction patterns obtained by pulsed-field gel electrophoresis. VTEC O118:H16/NM are emerging pathogens in Germany and should be classified as new enterohaemorrhagic E. coli (EHEC) types.  相似文献   

5.
The aim of the study was to define the prevalence of verotoxin-producing Escherichia coli (VTEC) in cattle and pigs in a Hong Kong abattoir. Faecal and carcass samples collected from 986 cattle and 487 pigs from an abattoir were tested for verotoxin (VT) by PCR and cytotoxicity assays. VTEC was isolated from 415 and 1-8% of cattle faecal and carcass samples and from 2.1 and 0.2% of porcine faecal and carcass samples, respectively. Amongst 409 VTEC isolates from cattle, 9 were serotype O157:H7 and eaeA+. The most prevalent vt genotype among bovine VTEC was vtl+vt2 (73.8%) and in porcine VTEC was vt2e+ (30%). None of the porcine VTEC isolates and 9.3% of the bovine VTEC isolates was eaeA+. The non-O157 serogroup VTEC isolates carrying eaeA and EHEC-hlyA belonged to serogroups O172, O15, O84, O91, O110 and O121. The local dietary preference for pork or chicken (rather than beef), the low VTEC carriage in pigs, the rarity of additional virulence factors (caeA) in VTEC isolated from cattle may explain the apparently low incidence of human diarrhoeal disease associated with VTEC in Hong Kong hitherto. However, the presence of non-O157 VTEC strains carrying the eacA virulence marker in cattle highlights the fact that sole reliance on sorbitol-MacConkey agar for screening human VTEC isolates may underestimate the human disease burden. The changing dietary habits of the population in Hong Kong reinforce the need for continued vigilance.  相似文献   

6.
Eight non-O157:H7 Vero cytotoxin (VT)-producing Escherichia coli (VTEC) strains isolated from ill persons and nine bovine and lamb strains of serogroups matching the human strains, were characterized for various properties known to be associated with E. coli virulence. Five different serogroups were represented: O5, O55, O103, O111 and O153. The bovine and lamb strains produced VT1, while 3 human strains produced VT1, 3 produced VT2 and 2 were positive for both VT1 and VT2. The strains were non-haemolytic on horse blood agar, did not produce either heat stable toxin A (STA) or heat labile toxin (LT), and were noninvasive. The CVD419 probe which has been proposed to identify enterohaemorrhagic E. coli (EHEC) hybridized with all of the O5 and O103 strains, none of the O55 and O153 strains, and 3 of the 4 O111 strains. The strains carried several different sized plasmids and hybridization of Southern blots with the CVD419 probe identified plasmids ranging in size from 42 x 10(6) to 90 x 10(6). The strains did not hybridize with the enteroadherence factor (EAF) probe derived from an enteropathogenic strain and associated with the ability to give localized adherence to HEp-2 cells. Nevertheless five of the strains adhered in a localized pattern to HEp-2 cells and Intestine 407 cells. Adhesion to either HEp-2 or Intestine 407 cells did not correlate with hybridization with the CVD419 probe or haemagglutinating properties.  相似文献   

7.
Virulence factors of Verotoxin-producing Escherichia coli(VTEC) strains isolated from hamburgers and ground beef were studied in Argentina by PCR. Their virulence profiles were correlated with those corresponding to strains isolated from calves and adult cattle. Most virulent profiles (VTs+ eae +Mp+) were present in E. colifrom healthy and diarrheic calves corresponding to O5:H-, O5:H27, O20:H?, O26:H11, O38:H?, O103:H-, O103:H2, O111:H-, O118:H16, O165:H-serotypes. The presence of the eaegene was significantly more frequent among VTEC strains isolated from calves (20/26; 76%) than from adult cattle (1/39; 2.5%) (p< 0.005). VT2+ eae ? E. coliwas prevalent in foods and adult cattle at slaughterhouse. The prevalence of the eaegene was similar between VTEC strains isolated from meat (0/21) and adult cattle (1/39; 2.5%) which constitutes the main population processed at slaughterhouses in Argentina. Serotyping showed that VTEC strains were distributed among 31 serotypes, some of which (O20:H19, O91:H21, O113:H21, O116:H21, O117:H7, O171:H2, OX3:H21) were shared between bovine and food strains. These O serogroups have been isolated from cases of haemorrhagic colitis (HC) and haemolytic-uraemic syndrome (HUS) in humans in several continental European countries. This study confirms the role of cattle as a reservoir of many VTEC serotypes other than O157:H7 and represents a base for future diagnostic, prevention and control strategies of EHEC in this country. In addition, this study affirms the advantages of PCR-based screening of E. coliisolates given the finding of so many verotoxin-producing strains.  相似文献   

8.
Two cohorts of 10 and 16 calves were followed at weekly or fortnightly intervals from 4-28 and 1-9 weeks respectively to determine whether natural infection by Vero cytotoxin (VT) producing Escherichia coli (VTEC) occurred. Ninety-one of 171 (53%) faecal specimens were VTEC positive and 20-80% of animals at any given time excreted VTEC. Of 104 VTEC strains studied further, 6 different serogroups (O 22.H16; O 25.H5; O 49.H-; O 86.H26; O 88.H25; O 153.H12) and an untypable strain (O? .H21) were identified. All strains belonging to the same serotype had identical profiles of reactivity with DNA probes to toxins VT1 or 2, LTI or II and a probe (CVD419) derived from a plasmid carried by enterohaemorrhagic Escherichia coli O 157.H7. Four of these serotypes were found in the faecal flora of the calves, taken as a group, throughout the 4-month study period. Sixty percent of the strains hybridized with the probe for VT1, 4% with the probe for VT2, and 36% with both probes. Faecal VTEC were significantly associated with overt diarrhoeal illness in animals < 10 weeks of age, but no characteristic profile of markers (serotype or hybridization pattern) in E. coli isolates was associated with diarrhoea. A serological response to VT1 was detected in some animals, but faecal VT1 VTEC excretion persisted in spite of seroconversion. VT1 seroconversion was not associated with diarrhoea. A serological response to VT2 was not detected even in those animals excreting VT2 VTEC in the faeces.  相似文献   

9.
This study determined the prevalence of the eaeA gene and its relationship to serotype and type of verotoxin produced in a collection of 432 verotoxigenic Escherichia coli (VTEC) obtained from the faeces of healthy cows and calves in a systematic random survey involving 80 dairy farms in Southwest Ontario. A PCR amplification procedure involving primer pairs which target the conserved central region of the O157:H7 eaeA gene showed that 151 (35.2%) strains were positive for the eaeA gene. All isolates (9-21 for each O group) of O groups 5, 26, 69, 84, 103, 111, 145 and 157 were positive, whereas all isolates (7-34 for each O group) of O groups 113, 132, and 153 and serotype O156:NM (38 isolates) were negative for eaeA. Seventy-three percent of 130 isolates of eaeA-positive serotypes produced VT1 only compared with 20% of 253 isolates of eaeA-negative serotypes. We conclude that there is a strong association between certain O groups and the eaeA gene, that serotypes of eaeA-positive and eaeA-negative VTEC implicated in human and cattle disease are present at high frequency in the faeces of healthy cattle, that VT1 is more frequently associated with eaeA-positive than with eaeA-negative serogroups, and that the eaeA gene is more frequently found in VTEC from calves compared with VTEC from adult cattle.  相似文献   

10.
An epidemiological study was carried out to determine the incidence and the serotypes of verotoxigenic Escherichia coli (VTEC) that cause infections in Galicia (north-western Spain). Although, VTEC strains were isolated from 55 (14%) of the 387 calves sampled and the majority of bovine VTEC strains belonged to serotypes (026:H11 or H–, 091:H21, 0103:H2, 0105:H18, 0111:H–, 0113:H21, 0126:H–, 0128:H– and 0157:H7 or H–) previously associated with human haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS) in other countries, VTEC are not a common cause of human infections in Spain. Thus, VTEC (026:H11 and 086:H10) were isolated from only 3 (0.6%) of the 482 children with diarrhoea investigated. We examined the 69 (3 humans and 66 bovines) VTEC strains that were initially isolated as E. coli producing a toxin cytotoxic to Vero and HeLa cells by polymerase chain reaction (PCR) using specific primers for VT1, VT2 and eae genes. PCR showed that 38 (55%) of VTEC strains carried VT1 genes, 18 (26%) possessed VT2 genes, and 10 (14%) carried both VT1 and VT2 genes. Three (one human and two bovine) strains which were formerly VTEC had lost the ability to produce verotoxins upon subculture and became negative for VT 1 and VT2 by PCR. In total 35 (51%) of 69 VTEC strains, including the two human VT1+ strains of serotype 026:H11, were positive for eae sequences when tested by PCR. Presence of the eae gene was significantly more frequent (100%; 21/21) among VTEC strains with serotypes (026:H11, 0111:H–, 0157:H–and 0157:H7) considered as enterohaemorrhagic E. coli (EHEC) than among VTEC strains with non-EHEC serotypes (29%; 14/48) (p < 0.001). Results obtained in this study indicate that cattle may be an important source of VTEC involved in human disease. However, severe clinical syndromes caused by VTEC, such as HC and HUS, are uncommon in Spain, in comparison with North America and the UK. In any case, VTEC disease can appear on the scene very suddenly, as occurred in the UK and North America in the 1980s.  相似文献   

11.
Some strains of Escherichia coli belonging to serogroups O26, O55, O111 or O128 produce Vero cytotoxin (VT). These serogroups are included in the range of enteropathogenic E. coli (EPEC) serogroups for which commercial antisera are available. In an attempt to obtain information on VT-producing strains other than those of serogroup O157, 122 strains belonging to these four serogroups and isolated in 1991 from patients with diarrhoea in the United Kingdom were tested for hybridization with VT probes. Only 18 of the 122 strains were VT-positive and these were O26 or O128. However 90 strains hybridized with the E. coli attaching and effacing (eae) probe (including 14 VT-positive strains) and 17 with the enteroaggregative E. coli (EAggEC) probe. For 78 eae-positive and 9 EAggEC-positive strains, tissue culture tests correlated with the probe results as the strains gave, respectively, either localized adhesion and a positive fluorescent-actin staining test or a characteristic aggregative attachment. A total of 111 of the 122 strains belonging to serogroups O26, O55, O111 or O128 possessed properties that may be associated with the ability to cause human diarrhoeal disease, and similar studies are needed on strains from the other classical EPEC serogroups.  相似文献   

12.
Faecal samples from 146 diarrhoeic lambs and goat kids, and from 511 healthy sheep and goats were screened for the presence of Vero cytotoxin-producing Escherichia coli (VTEC). In healthy sheep and goats, VTEC were isolated in 24.4 and 16.2% of the animals, respectively. Moreover, VTEC were detected in 3.1 and 5.9% of the diarrhoeic lambs and goat kids, respectively. These data suggest that VTEC seems not to be associated with diarrhoea in lambs and goat kids. Only four VTEC strains were eae-positive. The absence of the eae gene in most of these VTEC strains could indicate that these strains are less virulent for humans that the classical eae-positive enterohaemorrhagic E. coli types. However, almost half (42.9%) and 12.2% of VTEC strains isolated from healthy sheep and goats, respectively, belonged to serotypes associated with severe diseases in humans.  相似文献   

13.
Certain strains of verotoxigenic Escherichia coli (VTEC), and in particular those belonging to serogroup O157, cause attaching and effacing (AE) lesions of the host gut mucosa during pathogenesis. The mechanisms involved with bacterial attachment and the destruction of microvilli are determined by a cluster of genes within the LEE region, which also encode five secreted proteins. Sera from patients with antibodies to the lipopolysaccharide (LPS) of E. coli O157 and other VTEC were tested for antibodies to these secreted proteins. Twenty-one of 34 (62%) sera with antibodies to the lipopolysaccharide (LPS) of E. coli O157 also contained antibodies to one or more of the secreted proteins. Five of 12 sera containing antibodies to the LPS of a range of other VTEC serogroups also contained antibodies to 1 or more of the 5 secreted proteins, as did 16 of 70 (23%) sera from patients with haemolytic uraemic syndrome (HUS), haemorrhagic colitis (HC) or diarrhoea, but without bacteriological evidence of infection with VTEC and which did not contain antibodies to VTEC serogroups O5, O115, O145, O153 or O157. The detection of serum antibodies to secreted proteins may provide additional information for interpreting the results of established lipopolysaccharide-based VTEC serology.  相似文献   

14.
A study on the prevalence of the faecal carriage of Escherichia coli O157 in lambs was performed in the major slaughterhouse in Rome, central Italy, during 2002. A total of 643 animals, consisting of 378 weaned and 265 suckling lambs, were assayed for the presence of E. coli O157. Five O157-agglutinating E. coli strains were isolated (0.8%, 95% CI 0.3-1.9). Only one was positive to PCR specific for the eae gene and produced verocytotoxin VT2, with a VTEC O157 overall prevalence of 0.2% (95% CI 0.0-1.0), whereas one strain possessed the eae gene only. All the other isolates were negative for the presence of all the virulence genes considered. The animals were either from local farms or imported from Eastern Europe. The results suggest an age-specific difference since the microorganism was isolated only from 0.3% (95% CI 0.0-1.7) of weaned lambs, while all samples from suckling lambs tested negative. From this study, the overall risk of human exposure to pathogenic E. coli O157 from lamb meat consumption derived from the major slaughterhouse in Rome can be considered reasonably low, particularly when suckling lamb meat is considered.  相似文献   

15.
To investigate if there is a reservoir of Escherichia coli O157 in Norwegian cattle, faecal samples from 197 cattle herds were screened for E. coli O157 by the use of immunomagnetic separation (IMS) and PCR during the 1995 grazing season. Six E. coli O157:H-isolates were detected in two herds, one isolate in one and five in the other. The isolates carried the stx1, stx2, and eae genes, and a 90 MDa virulence plasmid. They were toxinogenic in a Vero cell assay. From 57 other herds, 137 faecal samples were positive for stx1 and/or stx2 genes detected by PCR run directly on IMS-isolated material. Among these samples, stx2 were the most widely distributed toxin encoding genes. No difference was found among milking cows and heifers in the rate of stx1 and/or stx2 in positive samples.  相似文献   

16.
Faecal samples of 2660 domestic animals from 116 farms and 956 samples of food were examined for the presence of Shiga toxin-producing Escherichia coli (STEC). STEC was recovered from 126 (15.3%) cattle, 135 (11.3%) pigs, 135 (66.8%) sheep, 31 (73.8%) goats, 4 (1%) chicken, and 15 (1.6%) food samples. Of all STEC isolates, 21.5, 25.8 and 15% produced enterohaemolysin, alpha-haemolysin, and aerobactin respectively, 1.6% displayed localized adherence (LA) to HEp-2 cells, 27.6% were sorbitol negative, and 30% were resistant to antibiotics. Only 14 (3.1%) of the STEC isolates belonged to human infection-associated serogroups (O26, O55, O111, O128 and 0157), designated as enterohaemorrhagic E. coil (EHEC). This study revealed that STEC are prevalent in domestic animals, and to a lesser extent in food of animal origin in Serbia, but the absence of a EHEC phenotypic profile (characteristic serogroup, LA, enterohaemolysin production) in most animal STEC strains may explain the low incidence of human STEC infection in this part of the world.  相似文献   

17.
Abstract A total of 52 Shiga toxin-producing Escherichia coli (STEC) strains, isolated from fecal samples of six ibex, 12 chamois, 15 roe deer, and 19 red deer were further characterized by subtyping the stx genes, examining strains for the top nine serogroups and testing for the presence of eae and ehxA. Eleven of the 52 strains belonged to one of the top nine STEC O groups (O26, O45, O91, O103, O111, O113, O121, O145, and O157). Eight STEC strains were of serogroup O145, two strains of serogroup O113, and one strain of serogroup O157. None of the strains harbored stx2a, stx2e, or stx2f. Stx2b (24 strains) and stx1c (21 strains) were the most frequently detected stx subtypes, occurring alone or in combination with another stx subtype. Eight strains harbored stx2g, five strains stx2d, three strains stx1a, two strains stx2c, and one strain stx1d. Stx2g and stx1d were detected in strains not harboring any other stx subtype. The eae and ehxA genes were detected in two and 24 STEC strains, respectively. Considering both, the serogroups and the virulence factors, the majority of the STEC strains isolated from red deer, roe deer, chamois, and ibex do not show the typical patterns of highly pathogenic STEC strains. To assess the potential pathogenicity of STEC for humans, strain isolation and characterization is therefore of central importance.  相似文献   

18.
Molecular techniques were used for studying the epidemiology of diarrhoeal infections due to Escherichia coli in the Gauteng region in South Africa. In total, 151 E. coli strains isolated from stools of patients with diarrhoea and 30 strains isolated from stools of healthy individuals were collected between March 1996 and May 1997. The E. coli isolates were characterized by serotyping, antimicrobial susceptibility testing, and adherence patterns. Polymerase chain reaction (PCR) was performed to determine the presence of the genes-encoding virulence factors. PCR showed that 59 (32.6%) of the E. coli isolates carried eaeA genes, 6 (3.3%) possessed bfpA genes, 4 (2.2%) CNF1, and 2 (1.1%) carried labile toxin and Stx2 genes. The eae genes were more prevalent in strains isolated from patients than in those from the control group (p < 0.001). Forty-eight (26.5%) strains belonged to enteropathogenic E. coli (EPEC) O serogroups and 14 (7.7%) to Shiga toxin-producing E. coli (STEC) O157 serotype. A high percentage (28.2%) of atypical EPEC strains possessing the eaeA but not the bfpA genes was isolated. Most isolates were susceptible to commonly-used antimicrobial agents. The adherence of the E. coli strains to HeLa cells was identified more in patients (69.4%) than in the control group (60%) and was more dominant in infants than in adults. PCR and tissue culture assays were shown to be useful techniques for the epidemiological study of E. coli where this organism is a major cause of diarrhoea.  相似文献   

19.
To assess the importance of infection by Verotoxin (VT) producing Escherichia coli (VTEC) in children with HUS in Central Europe, stool and/or serum samples obtained from 147 patients from 28 paediatric centres were prospectively examined for the presence of VTEC and the kinetics of faecal VT titres (FVT), and for VT neutralization titres and antibodies against E. coli O 157 lipopolysaccharide, respectively. Ninety-two percent of the patients had classic (enteropathic) HUS (E+ HUS). Evidence of VTEC infection was obtained in 86% of them. VTEC/FVT were identified in 55/118 E+ cases (47%). A prominent feature was the frequent isolation of sorbitol-fermenting, VT2-producing E. coli O 157.H-.VT1 (C600/H19) was neutralized by 9%, and VT2 (C600/933W) by 99% of the initial serum samples from E+ patients, compared to 3% (VT1) and 100% (VT2) from age-related controls. Fourfold titre rises against VT1 and/or VT2 were observed in 13/70 (19%), and significantly elevated O 157 LPS IgM and/or IgA antibodies in 106/128 (83%) of the E+ patients. The ubiquitous VT2 neutralizing principle in the serum of HUS patients as of healthy controls warrants further investigations.  相似文献   

20.
A prevalence study of verotoxin-producing Escherichia coli O157 (VTEC O157) was performed in 371 randomly selected dairy herds distributed throughout Sweden. Faecal and manure samples were collected and analysed by immunomagnetic separation and culturing. Data were recorded for each herd regarding herd size, age of sampled animals and whether, in addition to cattle, the farm kept other animals. VTEC O157 was isolated from 33 (8.9%) of the 371 investigated herds. The prevalence was higher (23.3%) in Halland county than in the rest of Sweden (P > 0.01). Halland was also the county in Sweden that during the study period had the highest incidence of human VTEC O157 cases. VTEC O157 was not detected on any farm in northern Sweden. Identified risk factors, in the multivariate analyses, for herds being VTEC O157 positive were herd size, geographical localization, presence of pigs on the farm and median age of sampled animals.  相似文献   

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