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1.
对5份恩施地区泡萝卜中的乳酸菌进行了分离鉴定,同时对其分离株在以萝卜为原料的泡菜中的发酵特性进行了评价。结果表明:分离出18株乳酸菌菌株,分别为隶属于片球菌属(Pediococcus)的戊糖片球菌(P.pentosaceus)和隶属于乳酸杆菌属(Lactobacillus)的食品乳杆菌(L.alimentarius)、短乳杆菌(L.brevis)、副干酪乳杆菌(L.paracasei)、发酵乳杆菌(L.fermentum)和植物乳杆菌(L.plantarum),其中8株分离株为L.plantarum。通过质构分析发现,乳酸菌纯种发酵制备的多数泡萝卜样品硬度和脆性均明显高于自然发酵样品。通过电子鼻分析发现,W1C、W3C和W5C对多数乳酸菌纯种发酵泡萝卜水的响应值明显偏高。通过主成分分析发现,菌株L.paracasei HBUAS51063和L.plantarum HBUAS51053具有相对较佳的发酵特性。由此可见,恩施市泡萝卜中乳酸菌以L.plantarum为主,乳酸菌纯种发酵可提升多数泡萝卜的品质。  相似文献   

2.
采用K-B药敏纸片法检测了5株具有潜在益生乳杆菌的耐药性,通过质粒消除,分析了菌株质粒与耐药性之间的联系,应用PCR确定了质粒决定的耐药基因.5株乳杆菌对万古霉素、多粘菌素B以及链霉素等7种抗生素普遍表现出抗性,但主要对四环素敏感.采用SDS与SDS-高温两种方法消除戊糖乳杆菌CH8的质粒后,菌株CH8表现出头孢噻吩和氯霉素敏感性.设计β-内酰胺类抗性基因blr、ECP-1569和nps-1以及氯霉素抗性基因cmlA、cat和cmlA1的引物进行PCR,与目的产物测序比对表明,戊糖乳杆菌CH8的质粒上含有β-内酰胺类抗性基因blr,该基因与其头孢噻吩抗性有关.该研究为探讨乳酸菌的抗药基因转移性提供了前期基础,有助于益生乳杆菌安全性评价体系的建立与完善.  相似文献   

3.
为得到适合黑果枸杞发酵的菌株,采用形态学观察、16s rDNA、26S rDNA与ITS全序列分析分离鉴定自然发酵黑果枸杞酵素中的乳酸菌和酵母菌,并对其安全性进行评价,包括药敏试验、溶血试验和急性毒性试验。结果表明:从酵素中分离得到了三株菌株分别为类食品乳杆菌(Lactobacillus paralimentarius)(M4),植物乳杆菌(Lactobacillus plantarum)(M6)和库德里阿兹威毕赤氏酵母菌(Pichia kudriavzevii)(Y5)。2株乳酸菌对青霉素类、头孢类、β-内酰胺类、大环内酯类及四环素类等大多数常见抗生素无耐药性;酵母菌Y5除了对氟康唑表现为耐药外,对其余抗真菌类抗生素均未表现出耐药性。3株菌均无急性毒性,也未发现有溶血现象。试验初步验证了菌株的安全性,为三株菌在黑果枸杞酵素产品的研发与生产中的应用提供了基础研究数据。  相似文献   

4.
对20株分离自山西老陈醋醋醅中的乳酸菌菌株进行益生特性和安全评价。结果显示植物乳杆菌172,173和发酵乳杆菌179具有较强的DPPH、胆固醇的清除能力,生物胺降解能力,ACE抑制活性及产细菌素、γ-氨基丁酸和乙偶姻的能力。毒力基因PCR结果显示大部分菌株呈阴性,仅有少量菌株呈现hyl(2/20)和ace(2/20)阳性。抗生素抗性基因erm B和tet M的检测率较高,分别达到70%(14/20)和65%(13/20)。另外2株淀粉乳杆菌显示出bla Z和van X阳性。有7株菌携带编码酪氨酸脱羧酶的tdc基因,5株菌携带odc(鸟氨酸脱羧酶)基因,未检测出hdc(组氨酸脱羧酶)基因。吲哚试验、硝酸盐还原酶、D-乳酸试验结果均为阴性。小鼠毒理实验进一步证明了植物乳杆菌172,173和发酵乳杆菌179这3株菌的安全性,为进一步研究与应用开发提供了科学依据。  相似文献   

5.
对恩施市酸豇豆中的乳酸菌进行了分离鉴定,同时使用电子鼻和电子舌技术对分离株纯种发酵制备酸豇豆的品质进行了评价。结果表明:分离出的23株乳酸菌菌株,共鉴定为Lactobacillus rhamnosus(鼠李糖乳杆菌),L. fermentum(发酵乳杆菌)和L. plantarum(植物乳杆菌)3个种,其中21株为L. plantarum。通过电子鼻分析发现,传感器W1C、W3C和W5C对多数L. plantarum纯种发酵的酸豇豆响应值明显偏高。通过电子舌分析发现,L. plantarum纯种发酵可明显提升酸豇豆的酸味。通过主成分分析发现,L. plantarum HBUAS51009、L. plantarum HBUAS51016和L. plantarum HBUAS51027纯种发酵的酸豇豆具有较好的品质。由此可见,恩施市酸豇豆中乳酸菌以L. plantarum为主,且添加L. plantarum进行纯种发酵可提升酸豇豆的品质。  相似文献   

6.
从酸奶中分离保加利亚乳杆菌,对其遗传多样性和药物敏感性进行分析,并进一步对耐药菌株的抗性基因进行检测。利用改良MRS培养基,从国内不同品牌酸奶中分离获得18株保加利亚乳杆菌。18株分离菌先经RAPD分型后采用琼脂稀释法测定其对11种抗生素的药敏性,并通过PCR对耐药菌株中可能存在的抗性基因进行检测。结果显示,18株受试菌具有明显的遗传多样性和耐药表型多样性。18株菌全部对罗红霉素敏感,而全部对卡那霉素耐药;对氨苄青霉素、青霉素G、金霉素、氯霉素、四环素、林克霉素、链霉素、新霉素及庆大霉素等9种抗生素均表现出不同程度的耐药性。通过检测耐药菌株的抗性基因,从1株菌(B-8)中检出四环素抗性基因tet(M),从2株菌(B-8和B-41)中检出链霉素抗性基因ant(6),从4株菌(B-43、B-47、B-49和B-51)中检出卡那霉素抗性基因aph(3’)-Ⅲa。结果表明,供试18株保加利亚乳杆菌多重耐药现象比较严重。  相似文献   

7.
从3种不同发酵周期的果蔬酵素中分离筛选得到15株菌种,通过16S rRNA基因序列、phe S基因和26S rRNA基因序列系统发育学分析,结合表型特征确定其分类学地位。结果表明:从6个月果蔬酵素中分离鉴定出菌种7株,含有2株类干酪乳杆菌(Lactobacillus paracasei)、1株植物乳杆菌(Lactobacillus plantarum)、2株毛榛毕赤酵母(Pichia manshurica)和2株嗜酒假丝酵母(Candida ethanolica);12个月果蔬酵素中分离出菌种5株,含有2株类干酪乳杆菌、1株短乳杆菌(Lactobacillus brevis)和2株毛榛毕赤酵母;18个月果蔬酵素中分离出酵母3株,均为毛榛毕赤酵母。与目前报道的菌种有异同,为酵素的工业化生产奠定菌种基础。  相似文献   

8.
为了研究湖北孝感和四川成都地区米酒曲中乳酸菌的多样性,该研究利用孝感凤窝米酒曲和成都农家自制米酒曲制作米酒, 并采用改良MRS固体培养基和分子生物学技术对米酒中的乳酸菌进行分离、鉴定。结果表明,从孝感米酒中共分离出42株乳酸菌,隶属于7个种,分别为戊糖片球菌(Pediococcus pentosaceus)、融合魏斯氏乳酸菌(Weissella confusa)、植物乳杆菌(Lactobacillus plan- tarum)、发酵乳杆菌(Lactobacillus fermentum)、能动乳杆菌(Lactobacillus agilis)、屎肠球菌(Enterococcus faecium)和乳酸乳球菌(Lac- tococcus lactis);从成都米酒中共分离出35株乳酸菌,隶属于7个种,分别为P. pentosaceus、W. confusa、L. plantarum、E. faecium、食窦魏 斯氏乳酸菌(Weissella cibaria)、约汉逊氏乳杆菌(Lactobacillus johnsonii)和短乳杆菌(Lactobacillus brevis)。两种米酒中分离出的相同菌种为P. pentosaceus、W. confusa、L. plantarum和E. faecium。其中,P. pentosaceus为两种米酒的优势菌株。  相似文献   

9.
以南美白对虾为研究对象,采用平板分离法从其肠道内筛得26株乳酸菌,通过16S rRNA测序鉴定可归为4种:乳酸乳球菌乳亚种L. lactis subsp. lactis、台湾乳球菌L. taiwanensis、格式乳球菌L. garvieae、乳酸乳球菌L. lactis。选取7株具有代表性的乳酸菌,用PCR的方法检测对虾中检出率较高的6类18种抗性基因(ARGs)在乳酸菌中的分布,辅以平板涂布的方法用抗生素选择性培养基研究乳酸菌对抗生素的耐药性。研究发现:7株乳酸菌具有相似的耐药谱,对四环素、磺胺吡啶、乙酰螺旋霉素表现出耐药性(抑菌率50.00%),对盐酸金霉、土霉素、硫酸庆大霉素、氯霉素、红霉素敏感(抑菌率100%);含有多重抗性基因,ARGs的检出率:磺胺类(92.86%)四环素类(53.06%)喹诺酮类(23.81%)氨基糖苷类(4.76%)氯霉素类=大环内酯类(0.00%),其ARGs基因型与菌株的抗生素表型并不能完全吻合。  相似文献   

10.
对湖北保康地区鲊广椒中的乳酸菌进行分离鉴定,并对其在玉米碜和红辣椒为原料的基质中的发酵特性进行评价。结果表明:分离出的20 株乳酸菌菌株,共鉴定为短乳杆菌(Lactobacillus brevis)、食品乳杆菌(L. alimentarius)、面包乳杆菌(L. crustorum)和植物乳杆菌(L. plantarum)4 个种,其中15 株为植物乳杆菌。对比自然发酵的鲊广椒,电子鼻传感器W1C、W3C和W5C对多数植物乳杆菌制备样品的响应值明显偏大,同时酸味、鲜味和丰度亦有明显的提升。通过主成分分析发现,使用L. plantarum HBUAS52327和L. plantarum HBUAS52332纯种发酵的鲊广椒具有优良的品质。由此可见,保康地区鲊广椒中乳酸菌以植物乳杆菌为主,且添加植物乳杆菌进行纯种发酵可提升鲊广椒的品质。  相似文献   

11.
The transfer via the food chain from animals to humans of microbes that are resistant to antimicrobial agents is of increasing concern. To determine the contributions of nonpathogenic microflora to the occurrence and spread of antibiotic resistance (AR) genes in the food chain, 123 lactic acid bacteria were isolated from 29 samples of raw and processed pork and chicken meat products that had previously tested positive for one or more AR genes that encode clinically relevant ARs: tet(M), tet(O), tet(K), erm(A), erm(B), erm(C), aac (6')-Ie aph (2")-Ia, mecA, and blaZ. All of the isolates were initially tested for their AR gene profiles by PCR. The 59 isolates carrying a tet, erm, or blaZ gene were taken through molecular identification, analyzed by determination of the MIC, and subjected to genetic fingerprinting. Lactococcus garvieae was the predominant species (28 isolates), followed by Lactobacillus plantarum (11 isolates) and L. salivarius (6 isolates), whereas Lactococcus lactis subsp. lactis, Lactobacillus johnsonii, L. reuteri, L. crispatus, and L. brevis were identified at lower frequencies. The tet(M) and erm(B) genes were the most frequently detected. Assessment of multiple resistances in 18 tet positive (tet+) isolates revealed that tet(M) plus erm(B) and tet(K) plus erm(B) were the most frequent AR gene patterns. Partial sequencing of the tet(M) open reading frame of three selected strains showed high sequence similarities (> 99%) with tet(M) genes previously found in human pathogens (Listeria monocytogenes and Neisseria meningitidis). Southern hybridization with plasmid profiles revealed these strains contained tet(M)-carrying plasmids.  相似文献   

12.
Susceptibility to 12 antibiotics was tested in 75 unrelated lactic acid bacteria strains of wine origin of the following species: 38 Lactobacillus plantarum, 3 Lactobacillus hilgardii, 2 Lactobacillus paracasei, 1 Lactobacillus sp, 21 Oenococcus oeni, 4 Pediococcus pentosaceus, 2 Pediococcus parvulus, 1 Pediococcus acidilactici, and 3 Leuconostoc mesenteroides. The Minimal Inhibitory Concentrations of the different antibiotics that inhibited 50% of the strains of the Lactobacillus, Leuconostoc and Pediococcus genera were, respectively, the following ones: penicillin (2, < or =0.5, and < or =0.5 microg/ml), erythromycin (< or =0.5 microg/ml), chloramphenicol (4 microg/ml), ciprofloxacin (64, 8, and 128 microg/ml), vancomycin (> or =128 microg/ml), tetracycline (8, 2, and 8 microg/ml), streptomycin (256, 32, and 512 microg/ml), gentamicin (64, 4, and 128 microg/ml), kanamycin (256, 64, and 512 microg/ml), sulfamethoxazole (> or =1024 microg/ml), and trimethoprim (16 microg/ml). All 21 O. oeni showed susceptibility to erythromycin, tetracycline, rifampicin and chloramphenicol, and exhibited resistance to aminoglycosides, vancomycin, sulfamethoxazole and trimethoprim, that could represent intrinsic resistance. Differences were observed among the O. oeni strains with respect to penicillin or ciprofloxacin susceptibility. Antibiotic resistance genes were studied by PCR and sequencing, and the following genes were detected: erm(B) (one P. acidilactici), tet(M) (one L. plantarum), tet(L) (one P. parvulus), aac(6')-aph(2") (four L. plantarum, one P. parvulus, one P. pentosaceus and two O. oeni), ant(6) (one L. plantarum, and two P. parvulus), and aph(3')-IIIa (one L. plantarum and one O. oeni). This is the first time, to our knowledge, that ant(6), aph(3')-IIIa and tet(L) genes are found in Lactobacillus and Pediococcus strains and antimicrobial resistance genes are reported in O. oeni strains.  相似文献   

13.
为鉴定感染黄颡鱼菌株fsznc-CL、建立一种快速检测的方法和探讨其耐药特性。采用形态学、理化特性及16S rDNA序列分析方法进行菌株鉴定,人工回归感染研究其致病性。通过对三对特异性引物的反应体系和条件进行优化,建立一种三重PCR检测法并应用到人工感染黄颡鱼的检测中。运用PCR方法扩增耐药基因,用琼脂纸片扩散法检测该菌的药敏特性。结果表明,分离菌株fsznc-CL为藤黄微球菌(Micrococcus luteus),该菌对黄颡鱼具有一定的致病性。三重PCR检测法可准确扩增出藤黄微球菌CBS、Sig和Pol三对目的基因,而其他菌株未扩增出,检测该菌的最低模板浓度为5.822×10-3 ng/μL,该方法检测带菌样品的阳性率约为83.33%。该菌携带了TEM、aph(3')-Ⅱa、aac(6')-Ⅰ、Sul1和Sul2耐药基因。菌株fsznc-CL对诺氟沙星、苯唑西林、红霉素和呋喃唑酮耐药。本试验建立的三重PCR检测法具有较高的特异性和灵敏度,目前藤黄微球菌对少部分抗生素耐药。  相似文献   

14.
Antibiotic resistance (AR) in bacteria, a major threat to human health, has emerged in the last few decades as a consequence of the selective pressure exerted by the widespread use of antibiotics in medicine, agriculture and veterinary practice and as growth promoters in animal husbandry. The frequency of 11 genes [tet(M), tet(O), tet(K), erm(A), erm(B), erm(C), vanA, vanB, aac (6')-Ie aph (2')-Ia, mecA, blaZ] encoding resistance to some antibiotics widely used in clinical practice was analysed in raw pork and chicken meat and in fermented sausages as well as in faecal samples from the relevant farm animals using a molecular approach based on PCR amplification of bacterial DNA directly extracted from specimens. Some of the 11 AR genes were highly prevalent, the largest number being detected in chicken meat and pig faeces. The genes found most frequently in meat were tet(K) and erm(B); vanB and mecA were the least represented. All 11 determinants were detected in faecal samples except mecA, which was found only in chicken faeces. erm(B) and erm(C) were detected in all faecal samples. The frequency of AR genes was not appreciably different in meat compared to faecal specimens of the relevant animal except for vanB, which was more prevalent in faeces. Our findings suggest that AR genes are highly prevalent in food-associated bacteria and that AR contamination is likely related to breeding rather than processing techniques. Finally, the cultivation-independent molecular method used in this work to determine the prevalence of AR genes in foods proved to be a rapid and reliable alternative to traditional tools.  相似文献   

15.
The objective of the study was to answer the question of whether the ready‐to‐eat meat products can pose indirect hazard for consumer health serving as reservoir of Enterococcus strains harboring tetracyclines, aminoglycosides, and macrolides resistance genes. A total of 390 samples of ready‐to‐eat meat products were investigated. Enterococcus strains were found in 74.1% of the samples. A total of 302 strains were classified as: Enterococcus faecalis (48.7%), Enterococcus faecium (39.7%), Enterococcus casseliflavus (4.3%), Enterococcus durans (3.0%), Enterococcus hirae (2.6%), and other Enterococcus spp. (1.7%). A high percentage of isolates were resistant to streptomycin high level (45%) followed by erythromycin (42.7%), fosfomycin (27.2%), rifampicin (19.2%), tetracycline (36.4%), tigecycline (19.9%). The ant(6′)‐Ia gene was the most frequently found gene (79.6%). Among the other genes that encode aminoglycosides‐modifying enzymes, the highest portion of the strains had the aac(6′)‐Ie‐aph(2′′)‐Ia (18.5%) and aph(3′′)‐IIIa (16.6%), but resistance of isolates from food is also an effect of the presence of aph(2′′)‐Ib, aph(2′′)‐Ic, aph(2′′)‐Id genes. Resistance to tetracyclines was associated with the presence of tetM (43.7%), tetL (32.1%), tetK (14.6%), tetW (0.7%), and tetO (0.3%) genes. The ermB and ermA genes were found in 33.8% and 18.9% of isolates, respectively. Nearly half of the isolates contained a conjugative transposon of the Tn916/Tn1545 family. Enterococci are widely present in retail ready‐to‐eat meat products. Many isolated strains (including such species as E. casseliflavus, E. durans, E. hirae, and Enterococcus gallinarum) are antibiotic resistant and carry transferable resistance genes.  相似文献   

16.
17.
Antibiotic resistance of lactic acid bacteria isolated from Chinese yogurts   总被引:2,自引:0,他引:2  
The aim of this study was to evaluate the susceptibility of 43 strains of lactic acid bacteria, isolated from Chinese yogurts made in different geographical areas, to 11 antibiotics (ampicillin, penicillin G, roxithromycin, chloramphenicol, tetracycline, chlortetracycline, lincomycin, kanamycin, streptomycin, neomycin, and gentamycin). The 43 isolates (18 Lactobacillus bulgaricus and 25 Streptococcus thermophilus) were identified at species level and were typed by random amplified polymorphic DNA analysis. Thirty-five genotypically different strains were detected and their antimicrobial resistance to 11 antibiotics was determined using the agar dilution method. Widespread resistance to ampicillin, chloramphenicol, chlortetracycline, tetracyclines, lincomycin, streptomycin, neomycin, and gentamycin was found among the 35 strains tested. All of the Strep. thermophilus strains tested were susceptible to penicillin G and roxithromycin, whereas 23.5 and 64.7% of Lb. bulgaricus strains, respectively, were resistant. All of the Strep. thermophilus and Lb. bulgaricus strains were found to be resistant to kanamycin. The presence of the corresponding resistance genes in the resistant isolates was investigated through PCR, with the following genes detected: tet(M) in 1 Lb. bulgaricus and 2 Strep. thermophilus isolates, ant(6) in 2 Lb. bulgaricus and 2 Strep. thermophilus isolates, and aph(3')-IIIa in 5 Lb. bulgaricus and 2 Strep. thermophilus isolates. The main threat associated with these bacteria is that they may transfer resistance genes to pathogenic bacteria, which has been a major cause of concern to human and animal health. To our knowledge, the aph(3')-IIIa and ant(6) genes were found in Lb. bulgaricus and Strep. thermophilus for the first time. Further investigations are required to analyze whether the genes identified in Lb. bulgaricus and Strep. thermophilus isolates might be horizontally transferred to other species.  相似文献   

18.
为探讨酸奶中乳酸菌所携带耐药基因对人类健康的潜在影响,对市售酸奶中的乳酸菌进行分离和鉴定,并通过药物敏感性实验确定菌株的耐药谱,同时利用聚合酶链式反应(polymerase chain reaction,PCR)扩增技术调查链霉素、庆大霉素、磺胺类和四环素等耐药基因的分布情况。结果表明:25 份市售酸奶样品中共分离得到56 株乳酸菌,包括26 株德氏乳杆菌保加利亚亚种、3 株植物乳杆菌、2 株嗜酸乳杆菌以及25 株嗜热链球菌。药敏结果显示,31 株乳杆菌对链霉素(87.1%)、庆大霉素(80.6%)、环丙沙星(74.2%)和四环素(61.3%)的耐药率较高,对头孢菌素类则较为敏感;而25 株嗜热链球菌同样对链霉素的耐药率最高,达76.0%;其次分别为万古霉素(32.0%)、环丙沙星(32.0%)和四环素(20.0%)。56 株乳酸菌中共检出5 种不同的耐药基因,分别为链霉素耐药基因ant(6)(检出率1.8%)、庆大霉素耐药基因aac(6')-aph(2')(检出率7.1%)、四环素耐药基因tetM(检出率5.4%)以及磺胺类耐药基因sulⅠ(检出率14.3%)和sulⅡ(检出率1.8%)。受试的乳酸菌中共有13 株检出耐药基因,其中有4 株携带两种不同的耐药基因。长期以来被认为安全并广泛应用于发酵食品领域的乳酸菌可能成为潜在的耐药基因贮存库。  相似文献   

19.
本文以传统发酵食品中分离的38株乳酸菌(lactic acid bacteria,LAB)为研究对象,利用溶血试验、明胶液化试验和吲哚试验测试受试菌株相关产毒代谢产物,采用K-B纸片法检测菌株对25种抗生素的耐药表型,运用聚合酶链式反应检测菌株携带的耐药基因。结果表明:38株乳酸菌的溶血试验、明胶液化试验和吲哚试验均为阴性,说明菌株在生长代谢过程中不产生相关毒力物质;38株菌对头孢曲松、头孢噻肟、四环素、多西环素、氯霉素、克林霉素和红霉素表现敏感;对其余18种抗生素呈不同程度耐受,其中对磺胺异噁唑耐药率最高(52.63%),其次是卡那霉素(34.21%)、万古霉素(31.58%)、阿米卡星(26.32%)、链霉素(15.79%)、利福平(2.63%);耐药谱分析受试大部分乳酸菌存在多重耐药现象;PCR共检测出3种耐药基因,分别是氯霉素耐药基因cat A(2.63%)、万古霉素耐药基因van A(5.26%)、利福平耐药基因rpo B(42.11%)。结果表明从传统发酵食品中分离的38株乳酸菌不存在相关产毒代谢产物安全隐患,但对部分抗生素呈现出不同程度耐受,并且利福平耐药基因rpo B的检出率较高。本研究为传统发酵食品中乳酸菌安全评价体系的完善提供理论参考。  相似文献   

20.
Probiotic bacteria and starter cultures of Lactobacillus, Weissella and Bifidobacterium of African and European origins were studied and compared for their susceptibility to antimicrobials. The study included, for all isolates, determination of MICs (Minimal Inhibitory Concentration) for 24 antimicrobials, detection of resistance genes by PCR reactions using specific primers and sequencing of positive amplicons. The ability of Lb. reuteri from Africa to transfer the erythromycin resistance gene erm(B) to closely related bacteria was investigated by conjugation. Variations were observed and high levels of intrinsic resistance were found among the tested species. Positive amplicons were obtained for resistance genes encoding aminoglycoside (aph(3')-III, aadA, aadE) and tetracycline (tet(S)) from isolates from Europe and macrolide (erm(B)) from an isolate from Africa. However, only the erm(B) gene found in Lb. reuteri L4: 12002 from Africa contained a homologous sequence to previously published sequences. This gene could be transferred in vitro to enterococci. Higher prevalence of phenotypic resistance for aminoglycoside was found in isolates from Europe.  相似文献   

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