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1.
Four isolates of Listeria monocytogenes from food, human and environmental sources were grown separately in broth (pH 6.0 at 8 degrees C) under atmospheres of air, 100% N(2), 40% CO(2):60% N(2) or 100% CO(2). Exponential and stationary phase cells were harvested to determine if growth atmosphere and growth phase influenced this pathogen's ability to survive exposure to an acid environment coupled with proteolytic enzymes, and the activity of bile salts. In general, isolates were more resistant to the acid environment than the bile salts environment and stationary phase cells were significantly more resistant to both environments than exponential phase cells. Irrespective of prior growth atmosphere, none of the isolates when in exponential phase remained detectable following full exposure to the acid environment (110 min at 37 degrees C) or the bile environment (3 h at 37 degrees C). With the exception of one isolate grown under the atmosphere of 40% CO(2):60% N(2), all isolates when in stationary phase were detectable following full exposure to the acid environment but death rates varied significantly. Stationary phase cells of all isolates grown under 40% CO(2):60% N(2) and 100% CO(2) were highly susceptible to the bile salts environment: cells were not detectable after a 2-min exposure whereas stationary phase cells grown under air or 100% N(2) were recovered following full exposure to the bile environment. Survival curves were characterised by a population decline of at least 3 log(10)/ml (from an initial level of 7 log(10) CFU/ml) in the first 15 min; thereafter a constant population number of approximately 4 log(10)/ml was maintained over the remaining exposure period. No survival was observed when stationary phase cells of L. monocytogenes FRRB 2538 grown in air and 100% N(2) were subjected to the acid environment followed by immediate exposure to the bile salts environment. The results showed that growth atmosphere and growth phase could influence survival of this pathogen against conditions that imitate the extremes of the most important nonspecific defence mechanisms against microbial infection: the acid environment of the stomach coupled with the activity of proteolytic enzymes, and the activity of bile salts in the small intestine.  相似文献   

2.
A total of 147 lactic acid bacteria was isolated from two types of naturally fermented dry sausages at four different stages of the ripening process studied in order to select the most suitable strains according to their technological characteristics including probiotic properties and antimicrobial activity against food-borne pathogens. Identification of the isolates revealed that 90% were lactobacilli, 4% enterococci, 3% Pediococcus sp. and sporadic isolates of Weissella viridescens, Leuconostoc pseudomesenteroides, and Leuconostoc sp. The isolated strains of Lactobacillus sakei (49 isolates), Lactobacillus curvatus (24 isolates) and Lactobacillus plantarum (7 isolates) were further characterized. All strains could grow at 15?°C, whereas the majority of the strains was able to grow in the presence of 6.5% NaCl and on acetate agar. The enzymatic potential of the strains was evaluated using the API ZYM system. During in vitro investigations all strains exhibited high leucine and valine aminopeptidase activities and moderate acid phosphatase and phosphohydrolase activities. Some strains showed very weak lipolytic activity. The enzyme profiling is an important factor for selection of strains as starter cultures. A large majority of the strains tolerated 0.1% bile salts whereas 58% of Lactobacillus curvatus strains and all Lactobacillus plantarum strains were resistant to 0.3% bile salts. All Lactobacillus sakei strains and the majority of Lactobacillus curvatus and Lactobacillus plantarum strains exhibited an anti-listerial activity against three Listeria monocytogenes strains. A percentage of 75, 50 and 29% of Lactobacillus sakei, L. curvatus and L. plantarum strains, respectively, could inhibit two Staphylococcus aureus strains. The contribution of the selected strains to a possible inhibition of Listeria monocytogenes and S. aureus in situ on fermented meats would be of considerable interest to enhance the hygienic quality of these products.  相似文献   

3.
Five strains of lactobacilli were studied for their ability to remove cholesterol in vitro under conditions that mimic the human gastrointestinal tract. The highest assimilation of cholesterol was observed in media supplemented with oxgall and the lowest in the presence of taurocholic acid. Scanning electron micrographs showed that cholesterol was adhered to the cellular surface of lactobacilli cells. Resting and dead cells were able to remove cholesterol although in small amounts. Additionally, inhibition of cholesterol micelles formation was observed in the presence of bile salts. All strains were able to deconjugate bile salts, where higher deconjugation was observed in the presence of sodium glycocholate compared with other bile salts studied. All strains also exhibited bile salt hydrolase activity and most strains showed higher substrate specificity towards glycine-conjugated bile than towards taurine-conjugated bile. The results indicated that lactobacilli could remove cholesterol in vitro via various mechanisms, and may exert such hypocholesterolaemic effects in vivo.  相似文献   

4.
Bile salt hydrolysis is an important metabolic reaction in the bile salt metabolism of mammals. This reaction has a facilitating effect for bile salt excretion but can also be involved in various illnesses. In recent years interest has increased to use bile salt hydrolysis to influence the cholesterol metabolism of humans and farm animals. To understand the distribution and range of bile salt hydrolase activity in lactic acid bacteria, we screened more than 300 strains of the genera Bifidobacterium and Lactobacillus and the species Lactococcus lactis, Leuconostoc mesenteroides, and Streptococcus thermophilus. Results obtained for 273 strains showed that bile salt hydrolase activity is common in Bifidobacterium and Lactobacillus but absent in L. lactis, Leu. mesenteroides, and S. thermophilus. Nearly all bifidobacteria species and strains have bile salt hydrolase activity, whereas this activity can only be found in selected species of lactobacilli. A strong correlation can be observed between the habitat of a genus or species and the presence of bile salt hydrolase activity. Most often bile salt hydrolase activity is found in strains that have been isolated from the intestines or from feces from mammals--an environment rich in conjugated and unconjugated bile acids. Strains and species from other habitats like milk or vegetables--environments from which bile salts are absent--do normally not have bile salt hydrolase activity. In two independent assays, we established that bile salt hydrolase activity in bifidobacteria is, in general, much higher than in lactobacilli.  相似文献   

5.
Eleven strains of lactobacilli were screened for their bile salt deconjugation ability, bile salt hydrolase activity (BSH) and co-precipitation of cholesterol with deconjugated bile. Bile salt deconjugation as determined by the release of cholic acid showed that more cholic acid was liberated from the deconjugation of sodium glycocholate than sodium taurocholate, and Lactobacillus acidophilus strains had higher deconjugation ability than L. casei strains. BSH activity, as quantified by the amount of taurine or glycine liberated from conjugated bile salts, indicated that substrate specificity was more towards glycine-conjugated bile compared to taurine-conjugated bile. Co-precipitation of cholesterol with cholic acid was observed from deconjugation of both conjugated bile, with more cholesterol being precipitated upon deconjugation of sodium glycocholate than upon that of sodium taurocholate. Cholesterol co-precipitation with deconjugated bile increased with decreasing pH. L. acidophilus ATCC 33200, 4356 and 4962 and L. casei ASCC 1521 showed highest deconjugation ability and BSH activity towards bile mixtures that resemble the human bile, and may be promising candidates to exert beneficial bile deconjugation activity in vivo.  相似文献   

6.
BACKGROUND: Bile salt deconjugation by Lactobacillus strains is often closely linked to bile tolerance and survival of the strains in the gut and lowering of cholesterol in the host. The present study investigated the deconjugation of bile salts and removal of cholesterol by 12 Lactobacillus strains in vitro. The 12 strains were previously isolated from the gastrointestinal tract of chickens. RESULTS: The 12 Lactobacillus strains could deconjugate sodium glycocholate (GCA, 16.87–100%) and sodium taurocholate (TCA, 1.69–57.43%) bile salts to varying degrees, with all strains except L. salivarius I 24 having a higher affinity for GCA. The 12 Lactobacillus strains also showed significant (P < 0.05) differences in their ability to remove cholesterol from the growth medium (26.74–85.41%). Significant (P < 0.05) correlations were observed between cholesterol removal and deconjugation of TCA (r = 0.83) among the L. reuteri strains (C1, C10 and C16) and between cholesterol removal and deconjugation of TCA (r = 0.38) and GCA (r = 0.70) among the L. brevis strains (I 12, I 23, I 25, I 211 and I 218). In contrast, although L. gallinarum I 16 and I 26 and L. panis C 17 showed high deconjugating activity, there was no correlation between cholesterol removal and deconjugation of bile salts in these strains. CONCLUSION: The results showed that the 12 Lactobacillus strains were able to deconjugate bile salts and remove cholesterol in vitro, but not all strains with high deconjugating activity removed cholesterol effectively. Copyright © 2009 Society of Chemical Industry  相似文献   

7.
The survival of lactic acid bacterial strains from eight meat starter cultures under conditions similar to those in the gastrointestinal tract was determined. The conditions in stomach were simulated by using pH 1-5 phosphate-buffered saline. The conditions in small intestine were simulated by using MRS broth over a pH range 4-7 and two bile salt concentrations (0.15% and 0.30%). The survival capacity at pH 3 was strain dependent. 0.30% bile salts were critical for screening bile salt tolerant strains at pH 6. Strains of Lactobacillus sake (RM10) and Pediococcus acidilactici (P2) had the best survival capacities under acidic conditions and at the higher concentration of bile salts.  相似文献   

8.
Lactobacillus sakei 1 is a food isolate that produces a heat-stable antimicrobial peptide (sakacin 1, a class IIa bacteriocin) inhibitory to the opportunistic pathogen Listeria monocytogenes. Bacterial isolates with antimicrobial activity may be useful for food biopreservation and also for developing probiotics. To evaluate the probiotic potential of L. sakei 1, it was tested for (i) in vitro gastric resistance (with synthetic gastric juice adjusted to pH 2.0, 2.5, or 3.0); (ii) survival and bacteriocin production in the presence of bile salts and commercial prebiotics (inulin and oligofructose); (iii) adhesion to Caco-2 cells; and (iv) effect on the adhesion of L. monocytogenes to Caco-2 cells and invasion of these cells by the organism. The results showed that L. sakei 1 survival in gastric environment varied according to pH, with the maximum survival achieved at pH 3.0, despite a 4-log reduction of the population after 3 h. Regarding the bile salt tolerance and influence of prebiotics, it was observed that L. sakei 1 survival rates were similar (P > 0.05) for all de Man Rogosa Sharpe (MRS) broth formulations when tests were done after 4 h of incubation. However, after incubation for 24 h, the survival of L. sakei 1 in MRS broth was reduced by 1.8 log (P < 0.001), when glucose was replaced by either inulin or oligofructose (without Oxgall). L. sakei 1 was unable to deconjugate bile salts, and there was a significant decrease (1.4 log) of the L. sakei 1 population in regular MRS broth plus Oxgall (P < 0.05). In spite of this, tolerance levels of L. sakei 1 to bile salts were similar in regular MRS broth and in MRS broth with oligofructose. Lower bacteriocin production was observed in MRS broth when inulin (3,200 AU/ml) or oligofructose (2,400 AU/ml) was used instead of glucose (6,400 AU/ml). L. sakei 1 adhered to Caco-2 cells, and its cell-free pH-neutralized supernatant containing sakacin 1 led to a significant reduction of in vitro listerial invasion of human intestinal Caco-2 cells.  相似文献   

9.
该研究以一株丁酸梭菌(Clostridium butyricum)LXKJ-1为出发菌株,通过常压室温等离子体(ARTP)诱变得到的一株耐丁酸 钠突变菌株LXYB-2,采用牛津杯法比较菌株LXYB-2与菌株LXKJ-1的抑菌能力,同时采用活菌计数法对两株菌株的耐高温、人工胃 液、人工肠液及胆盐等抗逆性进行比较。 结果表明,两株菌株对9种常见致病菌均有较好的抑菌活性,除单核细胞增生李斯特氏菌和 金黄色葡萄球菌外,两株菌的抑菌活性差异显著(P<0.05)。 在高温90 ℃处理5 min时,两株菌耐受性差异显著(P<0.05);人工胃、肠 液耐受性实验结果无显著差异(P>0.05);在胆盐质量分数达到0.4%时,两株菌胆盐耐受性差异显著(P<0.05)。 优良菌株LXYB-2更 适合作为益生菌饲料添加剂产品应用于动物养殖业。  相似文献   

10.
The present study aimed to evaluate selected probiotic properties of Lactobacilli isolated from the water of submerged cassava fermentation. Following Lactobacilli isolation, isolates were screened for their antimicrobial activity. Acid and bile tolerances, bile salt hydrolase (Bsh) activity spectrum were assessed. Among the 113 isolates obtained, 16 showed a broad antimicrobial activity spectrum against the indicator microorganisms. From these 16, 12 were found acid and bile resistant. They hydrolyzed glycoconjugated or tauroconjugated bile salts. From the four bile Bsh genes screened, only Bsh-Lp1 was found in five isolates. They identified as Lactobacillus paraplantarum, Lactobacillus paracasei, Lactobacillus brevis, and Lactobacillus plantarum. Based on the principal component analysis, L. paracasei 62L, L. plantarum 85L and 86L were selected as the most promising strains. These results suggest that water from submerged cassava fermentation can be a source of Lactobacilli with high probiotic potential.  相似文献   

11.
Three commercial phage sensitive strains of Lactobacillus delbrueckii and spontaneous phage resistant mutants isolated from them were studied by focusing on their biological and probiotic features. After incubation in a simulated gastric solution (pH 2.0), viable cell counts decreased moderately, but a protective effect on strains was observed when milk was added. A limited resistance to bile was observed, while most of the strains tolerated lysozyme, and grew in the presence of bile salts and fermented prebiotics. Some strains showed high hydrophobicity values and β-galactosidase activity. The strong antibacterial activity displayed toward pathogens was due to the production of lactic acid. Sensitive strains and their phage resistant variants were able to adhere to Caco-2/TC-7 monolayers and significantly inhibited the invasion of Salmonella enterica serovar Enteritidis into Caco-2/TC-7 cells. All strains tested showed potential probiotic features, suggesting a potential probiotic role of L. delbrueckii.  相似文献   

12.
Five strains of bifidobacteria were screened for their bile salt deconjugation ability, bile salt hydrolase (BSH) activity and co-precipitation of cholesterol with deconjugated bile. Bile salt deconjugation was determined by the release of cholic acid. All strains exhibited deconjugation of both sodium glycocholate and sodium taurocholate. More cholic acid was liberated from the deconjugation of sodium glycocholate than sodium taurocholate. BSH activity was quantified by determining the amount of glycine or taurine liberated from conjugated bile salts by bifidobacteria strains. There was higher substrate specificity for glycine-conjugated bile compared to taurine-conjugated bile. Co-precipitation of cholesterol with cholic acid was observed from deconjugation of both sodium glycocholate and sodium taurocholate, and by all bifidobacteria strains studied. More cholesterol was precipitated with cholic acid when sodium glycocholate was used compared to sodium taurocholate. Increased cholesterol co-precipitation with deconjugated bile was observed with decreasing pH levels. Bifidobacterium infantis 17930 showed highest deconjugation ability and BSH activity towards bile mixtures that resemble the human bile, and may be a promising candidate to exert beneficial bile deconjugation activity in vivo.  相似文献   

13.
Serotype distributions of Listeria monocytogenes in clinical samples and foods often differ. It is unknown whether such differences reflect a variation in the virulence of strains or are due to other factors that are not directly related to the strains' ability to cause illnesses. Fifty-two food and eight clinical isolates of L. monocytogenes were obtained from France, Japan, and the United States. Their pathogenicity in nonimmunocompromised female ICR mice was determined by intraperitoneal (i.p.) injection of the mice with test strains at 10(8) to 10(9) CFU per mouse. Five mice were injected with each Listeria strain and observed for 5 days. Listeria isolates that caused at least one death in 5 days were considered pathogenic. Isolates that caused no deaths in 5 days were considered nonpathogenic. All strains except Listeria innocua and one L. monocytogenes serotype 4b strain (RM3-1) isolated from bovine raw milk were pathogenic to nonimmunocompromised mice. Three food isolates of L. monocytogenes serotype 1/2c were weakly pathogenic to nonimmunocompromised mice, killing a maximum of 50% of mice at 10(8) CFU. Strains with no pathogenicity or reduced pathogenicity were further tested for their pathogenicity to immunocompromised mice. Each strain was inoculated i.p. into five mice at 10(3) to 10(10) CFU per mouse. No deaths of immunocompromised mice inoculated with 10(8) CFU were observed, but 20 to 40% of the mice died when inoculated with 10(9) CFU of L. monocytogenes RM3-1. The three L. monocytogenes serotype 1/2c isolates were also weakly pathogenic to immunocompromised mice, with two of the three isolates killing < or = 60% of mice at doses of < or = 10(8) CFU. The hemolytic activity of the three weakly pathogenic serotype 1/2c isolates was similar to that of pathogenic strains. However, the nonpathogenic strain RM3-1 was not found to be hemolytic on horse blood agar. We have identified several L. monocytogenes strains with reduced virulence levels. Further characterization of such isolates may aid in understanding factors affecting the variation in virulence among strains.  相似文献   

14.
Ren J  Sun K  Wu Z  Yao J  Guo B 《Journal of food science》2011,76(9):M622-M628
In vertebrates, bile salt hydrolysis plays an essential role in fat metabolism. Bile salts are synthesized in the liver. And in the small intestine glycine and taurine are de-conjugated from bile salts by the enzyme bile salt hydrolase (BSH) from intestinal microbes. However, the mechanism of bile salt hydrolysis in Lactobacillus plantarum is still ambiguous. Four predicted bile salt hydrolase (bsh) genes from L. plantarum ST-III were cloned into Escherichia coli. The function of these genes was explored by overexpression. All 4 proteins that were studied showed activity against glycine- or taurine-conjugated bile salts. Substrate preference was also observed in BSH proteins, especially for the enzyme BSH1, which had high hydrolysis activity for glycodeoxycholic acid. These results suggest that all 4 bile salt hydrolases may be responsible for the bile salt hydrolysis activity in L. plantarum ST-III. PRACTICAL APPLICATION: Hypercholesterolemia is considered one of the major risk factors for coronary heart disease. More interest has focused on intestinal microbes because of their role in the decrease of serum cholesterol. BSH proteins play an important role in the reduction of cholesterol. This paper adds to a better understanding of BSH proteins of intestinal microbes. It gives a great hint that probiotics can be used to solve hypercholesterolemia one day.  相似文献   

15.
ABSTRACT:  Eight strains of probiotic bacteria, including Lactobacillus rhamnosus , Bifidobacterium longum, L. salivarius, L. plantarum , L. acidophilus , L. paracasei , B. lactis type Bl-O4, and B. lactis type Bi-07, were studied for their acid, bile, and heat tolerance. Microencapsulation in alginate matrix was used to enhance survival of the bacteria in acid and bile as well as a brief exposure to heat. Free probiotic organisms were used as a control. The acid tolerance of probiotic organisms was tested using HCl in MRS broth over a 2-h incubation period. Bile tolerance was tested using 2 types of bile salts, oxgall and taurocholic acid, over an 8-h incubation period. Heat tolerance was tested by exposing the probiotic organisms to 65 °C for up to 1 h. Results indicated microencapsulated probiotic bacteria survived better ( P < 0.05) than free probiotic bacteria in MRS containing HCl. When free probiotic bacteria were exposed to oxgall, viability was reduced by 6.51-log CFU/mL, whereas only 3.36-log CFU/mL was lost in microencapsulated strains. At 30 min of heat treatment, microencapsulated probiotic bacteria survived with an average loss of only 4.17-log CFU/mL, compared to 6.74-log CFU/mL loss with free probiotic bacteria. However, after 1 h of heating both free and microencapsulated probiotic strains showed similar losses in viability. Overall microencapsulation improved the survival of probiotic bacteria when exposed to acidic conditions, bile salts, and mild heat treatment.  相似文献   

16.
Searches of the genome annotation of Listeria monocytogenes F2365, an isolate from the 1985 listeriosis epidemic in California, showed that this strain carries 20 authentic mutations resulting in premature stop codons, including a nonsense mutation in inlB. Here we showed that L. monocytogenes F2365 demonstrates atypical virulence-associated characteristics, including significantly (P < 0.05) reduced invasion efficiency in Caco-2 cells as compared with a closely related lineage I serotype 4b strain as well as significantly (P < 0.05) greater variation in invasiveness when grown under different conditions compared with standard laboratory control and other lineage I serotype 4b strains. In addition, L. monocytogenes F2365 demonstrated distinct growth characteristics, including a significantly (P < 0.05) reduced exponential growth rate when compared with laboratory control and other lineage I serotype 4b outbreak-associated strains as well as a significantly (P < 0.05) longer lag phase duration time compared with another lineage I serotype 4b strain. Our results support that L. monocytogenes F2365 is characterized by genotypic and phenotypic properties that are atypical of other L. monocytogenes strains.  相似文献   

17.
Deconjugation by bile salts hydrolases in probiotics has been related to reduction of serum cholesterol levels in mammals. We compared the susceptibility to conjugated primary (glycocholate and taurocholate) and secondary (glycodeoxycholate and taurodeoxycholate) salts and the level of hydrolase activity of Bifidobacterium strains with acquired resistance to bile and of their more sensitive original strains. Minimum inhibitory concentrations against conjugated salts of the resistant strains were higher than that of the corresponding originals. None of the strains displayed deconjugation against primary salts, whereas most of them deconjugated secondary salts. Salts of cholic acid were more toxic than that of deoxycholic acid. Derivatives showed higher hydrolase activity than their originals. These results suggested a relationship between bile resistance and deconjugation. Finally, the resistance of bifidobacteria against glycodeoxycholate increased in the presence of maltose and cellobiose as compared with glucose, which could be related to a more efficient energy procurement from disaccharides.  相似文献   

18.
L. monocytogenes represents a primary concern in the production of Gorgonzola, a Protected Designation of Origin (PDO) Italian blue-veined cheese produced only in the Piedmont and Lombardy regions. L. monocytogenes isolates (N=95) obtained from Gorgonzola rinds, paste, and production/ripening environments were serotyped and then genotyped using Pulsed Field Gel Electrophoresis (PFGE). The goal of this study was to investigate the variability of L. monocytogenes PFGE-types across different PDO Gorgonzola manufacturers (N=22). The majority of the strains (88%) were serotyped as 1/2a. PFGE identified 2 major pulse-types grouping 62 strains, detected from different plants and years, suggesting the presence of persistent and niche-adapted L. monocytogenes. In 9 plants, environmental strains shared the same pulse-types with strains from rinds or paste, suggesting a possible transmission pathway. Encouragingly, L. monocytogenes was retrieved from only 1 paste, indicating that production processes were under control in 21 plants. In the remaining plant, un-effective pasteurization or cross-contamination during production processes could be the cause of the contamination. Consequently, it is imperative that producers operate under the total respect of the Good Manufacturing Practices and following the principles of the Hazard Analysis Critical Control Point plans, in order to contain contamination throughout the whole processing.  相似文献   

19.
泡菜中乳酸菌的分离鉴定及抗性筛选   总被引:1,自引:0,他引:1  
该研究从泡菜中分离乳酸菌,通过形态观察及分子生物学技术对其进行鉴定,并对其进行人工胃液和胆盐耐受性试验,以期筛选性能优良乳酸菌。结果表明,从泡菜中共分离出71株乳酸菌,经鉴定分别为消化乳杆菌(Lactobacillus alimentarius)32株、植物乳杆菌(Lactobacillus plantarum)27株、发酵乳杆菌(Lactobacillus fermentium)1株、棒状乳杆菌(Lactobacillus coryniformis)2株、有害片球菌(Pediococcus damnosus)9株。其中,植物乳杆菌和发酵乳杆菌可用于食品,且植物乳杆菌S74和S78具有较高的抗人工胃液能力,存活率分别为(94.73±4.56)%、(108.73±7.16)%;菌株S74在0.3%的胆盐中的生长效率[(7.41±3.28)%]低于菌株S78[(10.04±4.90)%]。说明植物乳杆菌S78在pH 3.0的人工胃液和0.3%的胆盐环境均具有良好的耐受能力,在功能性泡菜及益生菌产品方面具有一定的开发潜力。  相似文献   

20.
The growth characteristics of meat processing plant-derived field strains of Listeria monocytogenes, L. welshimeri and L. innocua were analyzed. The strains were inoculated in BHI broth cultures and incubated at 4 and 7 degrees C. Growth curves were determined by colony counting for 28 days. Significant variations were detected in the growth properties of these field-derived strains. In particular some of the L. monocytogenes strains displayed better cold stress tolerance. These discrepancies in growth behavior were more apparent in the cultures at 4 degrees C compared to 7 degrees C. Similar growth characteristics were observed for selected L. monocytogenes strains also in food challenge tests based on a sliced bologna-type product. The results stress the need for more evaluation of field strain growth characteristics and incorporation of such information in relevant predictive microbial growth models for L. monocytogenes risk assessment in naturally contaminated food products.  相似文献   

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