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 46 毫秒
1.
The studies conducted in 2003–2004 focused on the possibilities of applying a tannic solution to remove adhesiveness from pikeperch eggs. Spawners were caught in Lake P?tnowskie (central Poland) and then transported to the Gos?awice Fish Farm. After initial selection, the fish were weighed, measured and stimulated with human chorionic gonadotropin. Gametes were obtained 5 days after the first injection. The weight and diameter of the eggs, and the commercial fecundity of individual females were determined. The eggs were fertilized with the dry method. After the addition of water, the eggs were mixed for 4 min, and then divided into 20 g portions. After determining the number of eggs in the various portions, the adhesiveness removal procedure was performed. Three concentrations of tannic acid solution (500, 1000, 1500 mg L?1) and three exposure times (0.5, 2, 5 min) were applied. The eggs were incubated in Weiss jars. The studies indicated that both the solution concentration and the exposure time significantly (P<0.05) impacted pikeperch egg hardening, the degree of adhesive removal and embryo survival. The tannic acid solution concentration of ≤500 mg L?1 applied for 0.5–2 min was not effective; the eggs clumped and it was impossible to separate them even with intensive mixing. Better results were obtained using higher tannic acid concentrations and/or by lengthening exposure time. The adhesiveness of pikeperch eggs disappeared completely after 5 min exposure to tannic acid solution concentrations of 500–1500 mg L?1 or after 2 min exposure to solution concentrations of 1000–1500 mg L?1. In these variants, the embryo survival rate to the eyed‐egg stage was 78.0–84.0% (2003) and 82.3–84.7% (2004). However, high tannic acid concentration had a negative impact on the pikeperch larvae hatching. The greatest decrease in survival rate was observed in groups exposed to a tannic acid solution of 1500 mg L?1 for 2 and 5 min periods. Thus, the optimum method for removing pikeperch egg adhesiveness was to apply a solution of tannic acid at a concentration of 500 mg L?1 for 5 min or 1000 mg L?1 for 2–5 min.  相似文献   

2.
Elimination of egg stickiness is an important factor in artificial reproduction of pikeperch (Sander lucioperca L.). This study was conducted to evaluate the effectiveness of Alcalase enzyme to remove the adhesive layer of pikeperch eggs. The eggs were treated with Alcalase at 0.5, 1.0, 1.5, 2.0 and 5.0 mL L?1 or a milk/talc solution 2 min post insemination. Duration of exposure was 2 min in Alcalase and 60 min in milk/talc. The highest, albeit not significant, hatching rate (85.4%) was found with 1.5 mL L?1 of Alcalase, but hatching rates were similar in 0.5, 1.0 and 2.0 mL L?1. Hatching rates were significantly lower groups treated with 5.0 mL L?1 Alcalase enzyme (56.4%) compared to groups treated with milk powder and talc (61.3%). Nominally complete removal of adhesiveness was observed in 1.5 and 2 mL L?1. All Alcalase treatments led to significantly lower incubation duration compared with the traditional milk/talc treatment. The application of Alcalase successfully eliminated pikeperch egg stickiness in less time than with traditional milk/clay/talc methods.  相似文献   

3.
《水生生物资源》2003,16(5):450-456
This study summarizes optimization of techniques for common carp artificial propagation including improvements of activation solution (AS), the process of insemination, and elimination of egg stickiness. The optimum gamete ratio for good fertilization and hatching rate ranged from 8490 to 23 672 spermatozoa per egg, when dechlorinated tap water was used. Optimal ratio between eggs (weight in g) and AS (in ml) was defined as 1:1 to 1:2. Different concentrations of AS such as NaCl from 0 to 34 mM (0–68 mOsmol kg–1) did not change fertilization and hatching rates. An AS adopted for carp spermatozoa (45 mM NaCl, 5 mM KCl, 30 mM Tris–HCl, pH 8) was compared with other saline AS; only the 51 mM (102 mOsmol kg–1) NaCl solution decreased fertilization and hatching rate. The AS containing 20 mM Tris–HCl at pH 9 increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or to AS of pH 6 and 7. Adhesiveness from the eggs was successfully removed by incubation in Alcalase DX (PLN 04715) using two successive steps with different enzyme concentrations. The first step with an enzyme concentration of 2 ml l–1 was applied from 8 to 20 min after fertilization. Later in a second step, the best time for application of alcalase enzyme at a concentration of 20 ml l–1 was for 45 and 60 s at 20 min after fertilization leading to fertilization and hatching rates of 80–87%. The α-Chymotrypsin (EC 3.4.21.1. Merck) was also found effective for elimination of stickiness. Results with α-Chymotrypsin enzyme indicate that the response to success in elimination of stickiness is highly variable mainly due to differences in the environment, quality of water and carp strains.  相似文献   

4.
Four desticking procedures for tench eggs (A: tannic acid solution (1 g L−1) for 15 s; B: alcalase enzyme solution (8 mL L−1) for 60 s; C: alcalase enzyme solution (15 mL L−1) for 120 s; D: Woynarovich and Woynarovich (1980) solution for 58 min followed by tannic acid solution (1 g L−1) for 15 s) were tested to obtain data about influence on embryo survival to hatching stage and further survival and growth of the larvae. In the tannic acid and Woynarovich and Woynarovich (1980) treatment (A and D) few eggs stuck together and some were adhered to the incubator walls, whereas in the alcalase treatments (B and C) eggs neither stuck together nor adhered to the incubator walls. Percentages of hatched larvae did not show significant differences (mean values ranged between 47.4% in treatment A to 37.0% in treatment C). Larvae deformities observed were <0.5% in all cases. There were no significant differences among survival and growth rates of the larvae from different egg desticking origin, reaching, after 30 days, mean survival values around 90% and total length and weight of 12.5 mm and 19 mg respectively.  相似文献   

5.
ABSTRACT:   Tropical minute rotifer strains (SS-type) induce mixis at 30–35°C but sexual reproduction and resting egg formation do not proceed well due to rapid environmental change. The present study examined the effect of temperature regulation on rotifer Brachionus rotundiformis (Langkawi strain, SS-type) resting egg formation in small (500 mL in culture volume)- and large-scale (500 L in culture volume) experiments. Rotifers were cultured at 30°C in 15–17 p.p.t. seawater with an initial density of 1 individual (ind.)/mL. After 4 days, when cultures were in exponential growth stage with active mixis induction, the culture temperature of the experimental rotifers was changed to 25°C. Control rotifers were cultured at 30°C throughout the experiment. Fresh or frozen Nannochloropsis oculata and condensed freshwater Chlorella vulgaris were used as the rotifer diets in the small- and large-scale experiments, respectively. Significantly higher resting egg production was observed with the experimental rotifers (30→ 25°C) versus the control rotifers. In the large-scale trial, experimental rotifers produced 2.6 × 106 resting eggs during a 9-day experiment, which was 1.6-fold more than the control rotifers. Moreover, the efficiency of resting egg formation was found to increase by a factor of 1.8. The present study indicates that decreasing culture temperature from 30 to 25°C after active mixis increased resting egg formation in B. rotundiformis (SS-type).  相似文献   

6.
Wolffish, Anarhichas lupus L. has internal fertilization. The eggs are released into water before the beginning of cleavage and stick together in clutches. To develop methods for separating eggs for subsequent incubation, changes during egg swelling and egg shell hardening were observed. Uninseminated eggs did not swell in ovarian fluid, but a perivitelline space formed in sea water because of a partial cortical reaction. In inseminated eggs kept in ovarian fluid, a cortical reaction took place and swelling was caused by elevation of the yolk membrane. Shrinkage of the yolk and absorption of water were observed after inseminated eggs were placed in sea water. Eggs swelled in ovarian fluid diluted by less than 30% sea water showed abnormal cleavage during subsequent development. An effective method for preventing egg stickiness after fertilization was to distribute them in trays with stagnant sea water for at least 5-6 h. Milk was effective for loss of egg adhesiveness at 50% concentration, but some eggs cleaved abnormally and subsequent mortality was high. The influences of ovarian fluid and milk on initial egg development with respect to peculiarities of egg swelling are discussed.  相似文献   

7.
ABSTRACT

An experiment was conducted to evaluate the effects of incubating pikeperch, Sander lucioperca, eggs in formalin and iodophor solutions for 15 min on embryo survival, the hatching rate, as well as on the rate of misshaped larvae, in order to develop methods for egg surface disinfection. Embryos in the morula stage, in the epiboly stage, and at the beginning of heart beat and blood circulation tolerated formalin concentrations up to 1,500 ppm for 15 min. However, they were very susceptible to iodophor treatment, as >0.1% iodophor solution (=13 ppm active iodine) significantly decreased the percentage of ready-to-hatch embryos and the percentage of hatched larvae. These data of this study recommend the use of formalin at a concentration of up to 1,500 ppm to disinfect pikeperch eggs.  相似文献   

8.
The elimination of egg stickiness is required for effective artificial reproduction of northern pike, but until now, available methods have required at least 40 min. Sodium hypochlorite was tested under laboratory conditions, and exposure to aqueous concentrations of 0.025–0.05% for 40 s effectively eliminated stickiness without adverse effects. Fertilization and hatching rates in laboratory trials were similar to those observed in eggs treated with traditional methods using clay or milk for 40 or 60 min, respectively, as well as those without treatment. Testing using conventional hatchery incubation techniques did not reveal differences in fertilization rates, while the number of hatched larva was significantly higher in eggs treated with sodium hypochlorite vs. clay. Eggs treated with sodium hypochlorite retained transparency, which facilitated monitoring of embryo development.  相似文献   

9.
Egg disinfection is considered the most important routine work in hatcheries to avoid fungal and/or bacterial infection of fish eggs. The aim of this study was to determine the effectiveness of three disinfectants: formalin, iodine and sodium chloride on the hatching success of yellow perch eggs. The disinfectants were tested in triplicate at different concentrations for 15 and 30 min bath treatments. Two experiments were conducted; formalin at five concentrations (25, 50, 100, 150 and 200 mg L?1) and 25 mg L?1 iodine were tested in the first experiment. The second experiment involved formalin at three concentrations (250, 500 and 1000 mg L?1), iodine at three concentrations (50, 100 and 250 mg L?1) and sodium chloride at three concentrations (500, 1000 and 3000 mg L?1) were used. Iodine and sodium chloride‐treated eggs hatched earlier than formalin‐treated eggs. The highest mean percentage of eyed stage, hatching rate and survival to first feeding fry was observed at 200 mg L?1 formalin for 30 min, 50 mg L?1 iodine for 15 min and 500 mg L?1 sodium chloride for 30 min. High concentrations of formalin (1000 mg L?1), iodine (250 mg L?1) and sodium chloride (1000 and 3000 mg L?1) showed toxicity to yellow perch eggs, resulting in low hatching rate and survival to first feeding fry. We recommended formalin at a concentration of 150–200 mg L?1 for 30 min as an effective, easily available and low‐cost disinfectant for routine use to improve yellow perch hatchability.  相似文献   

10.
Various procedures for artificial insemination in tench, Tinca tinca (L.) were re-examined with evaluation of fecundity of males and females among different tench strains. The objectives of this study were to enhance fertilization and hatching rates through optimization of the activation solution, the insemination process, the activation of gametes, and the elimination of eggs stickiness. Sperm for all experiments was collected directly into immobilization solution of modified Kurokura solution containing 180 mM of NaCl and stored at 2 °C for 2.5–5 h prior to the experiment. When dechlorinated tap water was used for activation a gamete ratio of 1150 spermatozoa per egg showed the best significant fertilisation and hatching rates. Optimal ratio between eggs (weight in g) and activation solution (in cm3) was 1:1. Different concentrations of activation solutions such as NaCl from 0 to 68 mM (0–136 mOsmol kg−1) without buffer statistically decreased fertilization and hatching rates. The activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 8 and 9 significantly increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 6 and 7. Adhesiveness of the eggs was successfully removed by incubation in Alcalase and activity: 3.16 Anson units per cm3.  相似文献   

11.
Studying gamete biology can provide important information about a species fertilization strategy as well as their reproductive ecology. Currently, there is a lack of knowledge about how long sea bass Dicentrarchus labrax eggs can remain viable after being activated in seawater. The objectives of this study were to understand the effects of pre‐incubation of fresh and overripe sea bass eggs in seawater and to determine the duration of egg receptivity. Pooled eggs (fresh and overripe) from four females were pre‐incubated in seawater for 0 min (control), 0.5 min, 1 min, 3 min, 10 min and 30 min and then fertilized by pooled sperm from four males. The fresh eggs had a higher fertilization success than overripe eggs. Our results revealed a significant effect of pre‐incubation time for both the fresh (P < 0.01) and overripe eggs (P < 0.01). Fertilization success of eggs significantly declined for both these treatments after 3 min of pre‐incubation, which clearly indicates that sea bass eggs are able to be fertilized by sperm for up to 3 min after release into seawater. This study has particular importance for understanding fertilization strategies, reproductive potential, as well as reproductive ecology of sea bass.  相似文献   

12.
Cultivated Atlantic cod (Gadus morhua) entering their first year of gamete maturation were fed diets with different levels of arachidonic acid (ARA) and eicosapentaenoic acid (EPA) for 6.5 months prior to commencement of spawning. Gravid females were stripped three times: at the beginning, peak and end of spawning. Lipid composition and egg and larval quality of 34 family crosses were investigated. Results indicated that ARA uptake into eggs from broodstock diet was highly efficient achieving proportions of ARA up to 84% higher in eggs than in the diet. EPA was 42–76% higher, and DHA was 155–173% higher in eggs than in diets. Cod fed the diet with the lowest EPA/ARA ratio had the greatest egg production. Eggs from fish on a diet with high ARA level had significantly higher fertilization and hatching success than those fed low levels of ARA. This diet produced on average 71 viable eggs g?1 female compared with 32.5 and 4 eggs in diet B and C, respectively. Furthermore, larval survival until 8 days posthatch was higher in diets with lower ARA levels. The combined results showed that ARA dietary supplementation and low EPA/ARA ratio yielded a greater number of viable larvae kg?1 female.  相似文献   

13.
Understanding the factors limiting copepod productivity in dense cultures is a prerequisite for the partial or entire replacement of Artemia and rotifers as live feed for finfish larvae. In dense cultures, high encounter rates between individuals may increase stress, cannibalism incidents and potentially trigger resting egg production. We conducted an experiment to evaluate the potential egg production and egg quality of Acartia tonsa stocked at densities ranging from 10 to >5000 ind. L?1. Egg Production (EP), Delayed Hatching Eggs production (DHE), hatching success (HS), egg mortality and water quality were used as end points. In the present system, A. tonsa was raised at >5000 ind. L?1 without affecting the mortality, confirming that attaining this high density in culture is possible. However, egg harvest reached an optimum of 12 000 egg L?1 day?1 at ~2500 ind. L?1 indicating that increasing stocking density above this level is not of practical interest. Calculations showed that the loss in egg harvest at stocking densities <2500 ind. L?1 is of 1.3% for every additional 100 adult copepods L?1. The increasing adult density did not affected the proportion of DHE produced (~10% of harvest) but decreased significantly the HS, though not to a point that would be problematic in a commercial production. Understanding the biology of copepods when stocked at high density is important to improve copepod culture systems and increase egg harvest yields. Technical solutions such as the continuous separation of eggs from adults in the water column, recirculation and the continuous provision of food are seen as potential solutions.  相似文献   

14.
去除卵膜及受精膜是成功制备棘皮动物染色体的关键步骤。为找到高效去除海胆卵膜与受精膜的方法,使用三氮唑(2 g/L)、盐酸海水溶液(pH 4.75)、二硫苏糖醇(3×10-3 mol/L)和对氨基苯甲酸(3×10-3 mol/L)等4种化学试剂分别对中间球海胆()卵子及各时期胚胎进行处理,比较了不同试剂、处理时间或处理时期的去膜率、去膜后受精率和胚胎畸形率等指标的差异;利用常规空气干燥法对经去膜处理的囊胚期胚胎制备染色体,并进行核型分析。结果表明,三氮唑、盐酸海水溶液、二硫苏糖醇和对氨基苯甲酸等4种试剂对中间球海胆的卵膜和受精膜均有去除效果,其中2 g/L三氮唑处理未受精卵30 min的去膜率为85.50%,去膜后受精率为97.25%,畸形率为1.75%,去膜效果优于其余处理组。利用经该方法去膜后的早期囊胚进行染色体制备效果较好,获得了61个分散良好、形态完整的染色体中期分裂相。核型分析表明,中间球海胆的二倍体染色体数为2n=42,核型公式为:2n=20m+20sm+2st,NF=84,即有10对中部着丝粒染色体,10对亚中部着丝粒染色体,1对亚端部着丝粒染色体,染色体臂数为NF=84。本研究可为海胆染色体制备及染色体操作育种提供技术参考。  相似文献   

15.
This study was conducted to determine the effects of broodstock diet on flame angelfish (Centropyge loriculus) reproduction and generate baseline spawning performance (mean fecundity, egg fertilization, egg viability) and egg biochemistry data for this species. Sixteen flame angelfish broodstock pairs were maintained on one of three formulated diets or a raw diet (consisting of squid, shrimp, fish eggs and algae) to determine the effect of diet composition on the fecundity, egg fertilization rates and egg viability over a 7‐month period. Although all the diets supported similar mean spawn frequency and fecundity over the duration of the experiment, the mean fertilization rates and egg viability were significantly greater among pairs receiving the raw diet. Egg fatty acid composition was generally reflective of the maternal diet and similar to the values reported for other tropical species. In particular, dietary factors such as n‐3 highly unsaturated fatty acid and ARA level (which were elevated in the raw diet and eggs from fish receiving that treatment) are correlated with egg quality and are therefore worthy of future investigation. Because of the ease of conditioning, adaptability to varied formulated diets and continuous, sustained reproductive output, flame angelfish are an excellent model for examining the factors affecting marine fish egg and larval quality.  相似文献   

16.
Batches of eggs (1 batch/female) from 17 rainbow trout in their first spawning season were stripped and reared separately, and the percentage which hatched and the percentage which reached the stage of first feeding were determined. The fertilised egg batches were analysed for egg wet weight, egg dry weight, chorion weight and levels in the eggs of free, bound and total lipid, precipitable protein, protein phosphorus, lipid phosphorus, calcium and iron. All determinations were made on single eggs, and six eggs from each batch were analysed for each variable. Results were expressed in absolute terms (as weight of component per egg) and as percentage of egg dry weight. Highly significant variations in all these aspects of egg composition were shown to occur between parent females. However, there was no significant correlation between the percentage of the original number of eggs which hatched and any one aspect of egg composition, except for a weak positive correlation (P<0.05) with the percentage of protein phosphorus in the eggs. Similarly, there was no significant correlation between the percentage of the original number of eggs which reached first feeding and any one aspect of egg composition except for weak positive correlations (P<0.05) with egg wet weight and with both the absolute level and the percentage of protein phosphorus in the egg. There was significant positive correlation (P<0.05) between the percentage of hatched eggs (alevins) surviving to first feeding and each of the following: egg wet weight, egg dry weight and absolute levels in the egg of bound lipid, precipitable protein and protein phosphorus. Egg batches with higher hatching percentage (>50%) differed significantly from those with zero hatching percentage in having (in absolute terms) higher egg weight, chorion weight, protein phosphorus (all P<0.001), egg dry weight, bound lipid and precipitable protein (all P<0.01) and (percentages) higher chorion weight (P<0.05) and protein phosphorus (P<0.001), and lower free and total lipid (both P<0.01) and iron (P<0.05). In a separate experiment to investigate the effects of allowing the eggs to be retained by the female within the abdominal cavity for increasing periods of time after ovulation, eggs were obtained from three females on three or four successive occasions 2–11 days apart. Although the above aspects of egg composition remained almost constant when the eggs were held in the female for up to 18 days after ovulation, the hatching percentage declined sharply within this period, in two females falling from over 90% to near zero. These results together indicate that the time of stripping of the eggs in relation to the date of ovulation is a much more significant determinant of egg quality than any of the chemical and physical aspects of egg composition which were investigated.  相似文献   

17.
Enzyme treatment to eliminate egg stickiness in tench, European catfish and common carp was compared with standard methodology in an attempt to decrease time consuming under hatchery conditions. Eggs of tench and E. catfish were exposed to an alcalase enzyme MERCK EC 3.4.21.14 (0.6 AU g?1) solution 3 min after egg fertilization for 2 min. The best enzyme concentration in tench and E. catfish was 10 and 20 ml l?1 of enzyme, respectively. The eggs of c. carp were successfully destickiness with ALCALASE DX (2.5 AU g?1) using two concentration of enzyme (2 ml l?1 and 20 ml l?1) from 8 to 20 min after fertilization.  相似文献   

18.
A continuous, underway fish egg sampler   总被引:2,自引:0,他引:2  
We describe a method to sample the highly contagious distribution of pelagic fish eggs. CUFES, the continuous, underway fish egg sampler, consists of a submersible pump, concentrator, electronics and sample collector. This system operates continuously and under nearly all sea conditions, providing a real-time estimate of the volumetric abundance of pelagic fish eggs at pump depth, usually 3 m. CUFES-derived estimates of volumetric abundance agree well with those from nets towed at pump depth and with areal abundance estimated from vertically integrated plankton tows. CUFES has been used successfully to sample the eggs of menhaden, pinfish, sardine, and anchovy off the coasts of the eastern and western United States and South Africa. Two large patches of eggs of the Atlantic menhaden were sampled off North Carolina in winter 1993–94, had a linear scale of 5–10 km, and were found in waters between the Gulf Stream and mid-shelf front. Spawning location may be related to bathymetry. CUFES is now being used to estimate spawner biomass by the daily egg production method. An optical plankton counter provided accurate estimates of the number of Atlantic menhaden eggs sample by CUFES.  相似文献   

19.
采用普通光镜、荧光显微镜和石蜡切片技术,对缢蛏受精和早期卵裂过程中的精子入卵、减数分裂、雌雄原核形成与结合、早期卵裂以及多精入卵等细胞学事件进行了显微观察。结果表明,缢蛏成熟未受精卵多呈圆球形或卵圆形,少数呈梨形,卵径为82~88μm,核相处于第一次成熟分裂中期;精子为典型的原生型,全长55~58μm,头部呈保龄球形,顶体前端的顶体杆呈特别细长的花丝状;在水温21~22℃、盐度10的条件下进行人工授精,精、卵混合后,精子迅速附着于卵子表面,启动卵子发育;受精后4~6 min,精子的头部已进入卵内并明显膨胀,卵子外形变圆,卵外附着的精子量明显减少;在受精后12~15 min、20~25 min,受精卵先后排出第一极体、第二极体,完成两次成熟分裂;第二次成熟分裂结束以后,精、卵核体积迅速膨胀,雄原核的膨胀早于雌原核,核膜重建,在受精后约30 min形成雌、雄原核;雌、雄原核均向卵子中央移动,雄原核旁边的精子星光清晰可见,随后二者在卵子中央以染色体联合的方式结合,联合核的染色体共同排列在纺锤体的赤道板上,形成第一次有丝分裂的中期分裂相;受精后40 min左右,受精卵进行第一次卵裂,染色体在纺锤丝的牵引下向两极移动,结果形成2个大小不等的卵裂球;受精后45~50 min,卵子进行第二次卵裂,核相变化与第一次卵裂相同,在与第一次卵裂垂直的纵轴方向上发生不等全裂,最终形成3小1大4个卵裂球;受精后60~70 min,胚胎进行第三次卵裂,仍为不等全裂,但自此次卵裂起已开始进行螺旋分裂。此外,对实验中发现的缢蛏极少量多精入卵、多极分离等异常细胞学现象进行了分析,并探讨了海洋贝类卵子阻止多精入卵发生的机制。  相似文献   

20.
Egg strings and nauplii of the salmon louse Lepeophtheirus salmonis were exposed to a variety of medicinal compounds at 50 mg L?1 for 30 min in two experiment series. This medicine concentration was selected as a starting point for a screening series. Hatching of egg strings and development to copepodid larvae were monitored in one experiment, and the survival and development of nauplii were monitored in the other. Two compounds, emamectin benzoate and cypermethrin, inhibited hatching effectively. Several compounds affected nauplii, either directly or through inhibiting development to the infective stage. A total of 50 mg L?1 of azamethiphos, cypermethrin, emamectin benzoate and propoxur was lethal to >70% of the larvae. Diflubenzuron, fenoxycarb, pymetrozine, pyriprole and tebufenozide diminished the ability of nauplii developing to copepodids.  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