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 influences of temperature and pH on the survival and growth of Saccharomyces cerevisiae, Candida stellata and Kloeckera apiculata were examined in the presence of ethanol concentrations between 2.5 and 15% v/v. At 15°C, the maximum concentrations of ethanol permitting the growth of S. cerevisiae, C. stellata and K. apiculata were 15%, 11% and 9%, respectively. These maximum concentrations were decreased at 10°C and 30°C. Cells of S. cerevisiae showed no loss in viability when incubated for 12 d at 10°C or 15°C in the presence of 15% ethanol but showed some loss at 30°C. Cells of C. stellata were tolerant of 12.5% ethanol at 10°C and 15°C but not at 30°C. Cells of K. apiculata were tolerant of 10–12.5% ethanol at 15°C but not at 10°C or 30°C. Sensitivity of the yeast cells to ethanol was marginally increased on decreasing the pH from 6-0 to 3–0.  相似文献   

2.
SUMMARY. Eggs of Ephemerella ignita (Poda) were kept at eight constant temperatures (range 5.9–19.8°C) in the laboratory. Over 85% of the eggs hatched in the temperature range 10.0–14.2°C but the percentage decreased markedly to 39% at 5.9°C and 42% at 19.8°C. Hatching time (days after oviposition) decreased with increasing water temperature over the range 5.9–14.2°C and the relationship between the two variables was well described by a hyperbola. Therefore, the time taken for development was expressed in units of degree-days above a threshold temperature. Mean values (with 95%CL) were 552 (534–573) degree-days above 4.25°C for 10% of the eggs hatched, 862 (725–1064) degree-days above 3.57°C for 50% hatched and 1383 (1294–1486) degree-days above 3.14°C for 90% hatched. These values can be used to predict hatching times at temperatures below 14.68°C for 10% hatched, 14.54°C for 50% hatched and 14.45°C for 90% hatched. At higher temperatures, the hatching time and the number of degree-days required for development both increased with increasing temperature. Equations were developed to estimate the number of degree-days required for development at these higher temperatures.
Eggs were also placed in the Wilfin Beck, a small stony stream in the English Lake District. Maximum and minimum water temperatures were recorded in each week and the summation of degree-days was used to predict the dates on which 10%, 50% and 90% of the eggs should have hatched. There was good agreement between these estimates and the actual hatching times. Only 10–15% of the eggs hatched between October and late February with most of the eggs hatching in March, April and May. Nymphs hatching in October and November probably did not survive the winter.  相似文献   

3.
CRYOPRESERVATION OF SEA URCHIN EMBRYOS AND SPERM   总被引:2,自引:0,他引:2  
A simple method for preserving live sea urchin embryos and sperm in liquid nitrogen (LN) wasdeveloped through the use of DMSO as a cryoprotective additive. Samples of late embryos in double test tubes were cooled to— I96°C by two-step freezing, first to — 76°C and then by plunging in LN. In the case of fertilized eggs, samples were previously frozen to — 40°C, at which temperature the samples were kept for 15 min; they were then transferred into LN. After preservation in LN for various lengths of time, samples in the double test tubes were thawed in water at 15°C. The post-thaw survival was more than 90% for late embryos, and about 10% for fertilized eggs. Difference in the length of the cryopreservation period did not affect survival. Post-thaw development in cryopreserved embryos often showed abnormalities in structure. Sperm with 1.5 M DMSO was successfully preserved in LN by a similar method to the one used for cryopreservation of late embryos. Fertilizability in cryopreserved sperm was complete, regardless of the length of the preservation period. Nearly all the eggs fertilized by cryopreserved sperm developed to normal plutei.  相似文献   

4.
Spermatozoa of the sea urchin, Hemicentrotus pulcherrimus (108 cells/ml), preincubated with unfertilized eggs deprived of jelly coats (more than l05 cells/ml) at 20°C for 20min in Mg2+ free artificial sea water containing 1 mM Ca2+ (MFASW), exhibited very low respiration, which was enhanced by 2, 4 dinitrophenol (DNP). The fertilization rate in MFASW was usually less than 5% and was about 25% at most. Preincubation with fertilized eggs (with and without a fertilization membrane) in MFASW did not reduced the respiratory rate of spermatozoa. The rate of sperm respiration was lower in MFASW than in artificial sea water (ASW), but was higher than the respiratory rate of spermatozoa preincubated in MFASW with unfertilized eggs. Sperm respiration in MFASW or in ASW was not stimulated by 2, 4 dinitrophenol. Almost complete inhibition of sperm respiration was obtained with unfertilized eggs fixed with glutaraldehyde at concentrations of above 105 cells/ml in MFASW and of about l04 cells/ml in ASW. The respiratory rate of spermatozoa treated with fixed eggs was enhanced by DNP. It is concluded that the respiratory rate of the spermatozoa is reduced by their interaction with unfertilized eggs before their penetration into the eggs.  相似文献   

5.
Grey mullet, Mugil cephalus , collected from ponds were induced to spawn with carp pituitary homogenate and human chorionic gonadotropin (HCG) at 30%o salinity and 26° C. The spontaneously spawned and fertilized eggs were transferred to sea water of different salinities in the range of 5–70%o, either at the 2-blastomere or the gastrula stage. Those eggs transferred at the gastrula stage were more tolerant to the salinity change than were those transferred at the 2-blastomere stage. The eggs did develop to the embryonic stage within the salinity range from 5 to 60%o. Hatching occurred in all salinities between 10 and 55%o. However, no larvae survived at 10 or at 55%o. The optimal salinity range for eggs incubating at 22.0–25.5° C was from 30 to 40%o salinity, with the peak at 35%o.  相似文献   

6.
The effects of various concentrations of sodium chloride solutions (0·1%–3%) and different temperatures (4, 10, 20, 30 and 37 °C) on survival of Legionella pneumophila were investigated. It was found that at temperatures between 4 °C and 20 °C, Legionella organisms survived in salt solutions up to 3% NaCl. Only the combination of high temperatures, i. e. 30 °C and 37 °C, with NaCl concentrations over 1·5%, reduced cell numbers significantly. It was interesting to note that the addition of small amounts of NaCl (0·1%–0·5%) enhanced survival of Leg. pneumophila , suggesting a protective effect of NaCl. In order to obtain information about conditions encountered in the environment, the survival experiments were repeated in sterile sea water from the Baltic Sea and the North Sea. The marked bacterial die-off, especially at higher temperatures, was not observed in natural sea water. All these results indicate that Leg. pneumophila can survive in the marine environment.  相似文献   

7.
The structure of the eggs of the sea urchin, Arbacia punctulata, has been investigated after the removal of one-half of the cellular protein. The procedure involves treatment of the eggs with 30 per cent ethanol at -10°C. followed by extraction of the soluble proteins with water. The eggs remain intact, although all of the cytoplasmic matrix is removed. Most cell structures can still be identified, although only the membranes of most remain. The mitochondria lose all of their matrix but retain the inner membranes or cristae. The annulate lamellae appear unaffected by this extraction procedure, remaining intact and apparently undamaged. The nuclear envelope is also retained, although it often undergoes a curious disorganization, apparently as the result of the separation of its two layers. The significance of these observations with respect to the structure of the envelope is discussed.  相似文献   

8.
ABSTRACT. In laboratory-laid eggs of the striped ground cricket, Allonemobius fasciatus DeGeer (Gryllidae), water absorption occurs at an early stage of embryogenesis (stage II) at 30°C but is delayed until later stages at lower temperatures. This is related to the variation in diapause stage at different temperatures. No egg developed beyond stage VII (the end of anatrepsis) without water absorption.
A. fasciatus shows seasonal variation in the stage of water absorption. At 30°C, eggs collected in August absorb water at the early stage while many of those collected in September avert diapause and absorb water at a later stage.
Diapause also influenced the water absorption of eggs in A. socius Scudder. Eggs of short-day females enter diapause and absorb water at stage II, while those of long-day females develop without diapause and absorb water at a later stage (around stage IV).
The susceptibility to desiccation (r.h. 50%) was examined at 20°C with A. fasciatus eggs. The percentage water loss and mortality of eggs varied with the time and duration of exposure to desiccation. Eggs are most sensitive to desiccation during the first several days after being laid and during the period of water absorption.  相似文献   

9.
In unfertilized eggs of the sea urchin Hemicentrotus pulcherrimus , fertilization membrane formation was induced by an incubation with dimethylsulfoxide (DMSO) for several min at 20°c followed by another incubation in an ice bath. The number of eggs with fertilization membrane, thus obtained, increased in relation to the concentration of DMSO between 1 and 3% (v/v) and was higher than 75% at concentrations above 3%. Fertilization membrane formation by this treatment occurred in Ca2+ free- or Ca2+, Mg2+ free- artificial sea water containing EGTA (50 mM) and was inhibited by verapamil. In the presence of DMSO, the membrane formation was also induced by 2, 4-dinitrophenol or cyanide in considerable number of eggs at 20°c. Eggs remained fertilizable, even when they were kept with DMSO for 1 hr at 20°c. DMSO slightly enhanced respiratory rate in unfertilized eggs and substantially reduced it in fertilized eggs. DMSO-treated eggs exhibited cyanide-insensitive respiratory burst following chilling in an ice bath or by adding DNP or cyanide, in a similar manner to the burst induced by sperm.  相似文献   

10.
Flushing of ballast tanks with seawater has been proposed to reduce the risk of invasion associated with residual ballast in 'no ballast on board' ships. The efficacy of this procedure, however, has not been determined. Using diapausing eggs isolated from ballast sediments — as well as from Lake Erie sediment — this study investigated the impact of salinity (0, 8 and 35‰) and temperature (10, 20 and 30 °C) on the cumulative abundance and species richness of hatched zooplankton taxa. The rate and amount of hatching varied dramatically between sediments and across salinity–temperature regimes. Although exposure to saline water inhibited emergence of freshwater taxa during the exposure phase of all trials, mixed results were evident after diapausing eggs were returned to freshwater. The efficacy of salinity as a ballast treatment method was temperature dependent, although the direction of the effect was case-specific. Exposure of eggs to saline water was less effective at 10 and 30 °C than at 20 °C. Although flushing ballast tanks with open ocean water is expected to significantly reduce the number of active invertebrates living in residual ballast water (a potentially larger source of invaders), our results indicate that the most effective treatment conditions for reduction of diapausing egg viability is 8‰ salinity at 20 °C.  相似文献   

11.
The effects of temperature on maintenance and termination of embryonic diapause were investigated in Jining (35.4°N, 116.6°E) and Sihong (33.5°N, 118.2°E) strains of the Chinese rice grasshopper, Oxya chinensis Thunberg (Orthoptera: Catantopidae). Eggs of both strains entered diapause when incubated at 30, 25, or 20 °C. Chilling at 8 °C had an evident effect on diapause termination and almost all eggs chilled for 60 days ended diapause development. Chilling of eggs at 8 °C for only 20 days failed to result in any hatching at 20 °C, suggesting that such level of chilling was not enough to induce diapause termination. However, the treatment combining incubation of eggs at 30 °C for varying lengths of time with subsequent incubation to 20 °C had a distinct effect on the completion of diapause of the eggs. The results indicate that there were two temperature optima, that is, low temperature (chilling) and high temperature, for diapause development in this grasshopper species. Incubation of chilled eggs at 20 °C for 5–15 days followed by further incubation at 25 °C reduced termination of diapause significantly compared with the eggs only chilled at 8 °C. Exposure of eggs chilled at 8 °C to a pulse of 25 °C from 1 to 7 days, separated by a 20-day interval at 8 °C, resulted in a decrease in the percentage of successfully hatched eggs as the length of the pulse of 25 °C increased. The results suggest that diapause intensity may be restored at moderately high temperatures. This reversible change in diapause intensity would play an important role in maintaining diapause before winter.  相似文献   

12.
Abstract. Embryonic diapause induction in the Senegalese grasshopper, Oedaleus senegalensis Krauss (Orthoptera: Acrididae), is influenced both by the photoperiod and the temperature experienced by females. High temperatures (40°C) and long photoperiods (LD 14:10h), which characterize the beginning of the rainy season in the Sahel, cause non-diapausing eggs to be laid. Lower temperatures (25°C) and shorter photoperiods (LD 12:12h), which occur at the end of the rains, result in the production of diapausing eggs. At 30°C and constant photoperiods, O. senegalensis exhibited a long-day-short-day response with critical photoperiods of c. 13 h and c. 20 h, only the former value being of ecological significance. The photoperiodically sensitive stages to diapause induction in females occurred from the fifth stadium onwards. Temperature also affected the coloration of both nymphs and adults. Dark-black and pale-white individuals were produced by low (25°C) and high (40°C) temperatures respectively, whereas an intermediate temperature (30°C) produced individuals which were greyish brown. These results are discussed in relation to the ecology of O. senegalensis.  相似文献   

13.
SUMMARY. 1. Egg survival (ES, percentage of eggs hatched in vitro ), reproductive success (RS, percentage of live young released from the brood pouch) and brood development lime ( d , days) in four populations of Gammarus fossarum and two populations of Gammarus roeseli were studied, in the laboratory at water temperatures of 2.0–26.1°C. Intraspecific differences between populations were not significant, but interspecific differences were found between the two species.
2. In natural stream populations, the reproductive period of G. fossarum lasted from December to September, that of G. roeseli from March to September.
3. In the experimental temperature range 2–26°C, 73% of the total number (771) of G. fossarum females and 69% of 469 G. roeseli females were ovigerous. Of these, 45% of G. fossarum and 43% of G. roeseli females successfully released live young from their brood pouches.
4. For G. fossarum , the optimum temperatures were 11.4°C for ES, where 76% of the eggs hatched, and 11.8°C for RS, where 77% of the females released live young from their brood pouches. For G. roseli , the optimum temperatures were 13.5°C for ES (51% hatched) and 14.0°C for RS (76% released). Over 50% of eggs hatched at temperatures of 3.6–19.2°C in G. fossarum and at 1 1.9–15.1°C in G. roeseli . Development time increased from 12 days at 21.9°C to 251 days at 2.0°C in G. fossarum , and from 10 days at 24.1°C to 212 days at 4.1°C in G. roeseli .
5. interspecific differences between the effects of water temperature on ES, RS and d are used to explain the different distributional patterns of G. fossarum and G. roeseli in central European running water systems. assuming that other physico-chemical variables are suitable for both species.  相似文献   

14.
Life history parameters of Amitus fuscipennis MacGown and Nebeker as parasitoid of Trialeurodes vaporariorum (Westwood) were determined at 15, 20, 25 and 30°C on tomato using three different methods. For each method, immature development, mortality, longevity, fecundity, oviposition frequency and post-oviposition period were determined and temperature-dependent relations were estimated. Oviposition frequency was also estimated as a function of parasitoid age. Immature development had a maximum of 61 days at 15°C that decreased to 22 days at 30°C. Mortality in the grey stage was less than 2% at temperatures lower than 30°C, where it was 60%. Longevity fluctuated between 3 and 18 days. Fecundity increased from 338 eggs/female at 15°C to a maximum of 430 eggs/female at 25°C and then decreased to 119 eggs/female at 30°C. Oviposition frequency varied between 3 and 46 eggs/female per day and had its maximum on the first day after emergence of the parasitoid. Net reproduction rate, generation time and intrinsic rate of increase were calculated. The intrinsic rate of increase increased from 0.090 at 15°C to a maximum of 0.233 at 25°C and then decreased to 0.159 at 30°C. The influence of the methods to determine the life history parameters on the results is discussed. The results are compared also with the life history of Encarsia formosa Gahan (Hym., Aphelinidae) a parasitoid of the same host. The advantages and disadvantages of the pro-ovigenic A. fuscipennis in comparison with the synovigenic E. formosa are discussed.  相似文献   

15.
Incubation of eggs of tuatara, Sphenodon punctatus   总被引:3,自引:0,他引:3  
Eggs of the tuatara, Sphenodon punctatus , were incubated either buried or half buried in vermiculite at constant temperatures of 15, 18, 20, 22 and 25 °C and constant water potentials between —90 and —400 kPa. Many clutches failed completely, possibly because they had been taken from females prior to proper shell development. Failed eggs were significantly smaller than successful eggs. Incubation is unsuccessful at 15 °C. Hatching success is high between 18 and 22 °C but low at 25 °C, but equally successful between 18 and 22°C. Incubation is strongly influenced by temperature, with mean incubation periods of 328 days at 18 °C, 259 days at 20 °C, 169 days at 22 °C and 150 days at 25 °C. Water potential generally has little influence on incubation time at a given temperature. Buried eggs hatch sooner than partially buried eggs at 20 °C but the large range makes significance dubious.
Eggs on the driest substrata at 18 and 20 °C lose water initially but then gain water through the rest of incubation. Eggs in all other conditions gain water throughout incubation, with the rate of i water absorption being maintained or increasing late in incubation. The suggestion that increasing rate of water absorption late in incubation facilitates explosive hatching is not supported. Egg mass at the time of hatching varies from 132 to 398% of initial values, depending on incubation conditions. Final egg mass is not affected significantly by incubation temperature. Hence, rates of absorption increase with temperature.
Water potential has no influence on hatchling size. However, hatchlings from buried eggs generally are significantly larger than those from partially buried eggs.  相似文献   

16.
The effects of temperature on the development and survival of Shijimiaeoides divinus barine were examined in the laboratory in 2008. The eggs and larvae were reared at temperatures of 15, 17.5, 20, 25, 30 and 35°C with a long-day photoperiod of 16 h light : 8 h dark (16L : 8D). The highest hatchability of eggs was 88.0% at 20°C, but hatchability at high temperatures of 30 and 35°C was 30 and 0%, respectively. The lowest and highest survival rates from the first to third instar were 18.8% at 15°C and 76.9% at 20°C. Few deaths were observed after the fourth instar. The shortest developmental periods of the eggs and larvae were 4.0 and 15.8 days at 30°C, and the durations of the egg and larval stages increased significantly as the temperature decreased. The developmental zero and thermal constants were 9.6°C and 82.6 degree–days for the egg stage, and 10.7°C and 306.8 degree–days for the larval stage. The developmental period of the natural population of S. divinus barine in Azumino City, Nagano Prefecture was calculated using the developmental zero, thermal constants and Azumino City temperature data.  相似文献   

17.
SUMMARY 1. The objective was to compare variations in egg hatching between the two species (interspecific variations) and between populations of the same species (intraspecific variations). There were significant interspecific, but not intraspecific, differences in female size, adult life-span, egg production, hatching success, incubation periods and hatching periods.
2. The optimum temperature for hatching success within the range 3.8–22.1°C in the laboratory and the range over which at least 50% of the eggs hatched were lower for Chloroperia tripunctata (Scopoli) (8.5°C, 4.2–17.3°C) than for Siphonoperla torrentium (Pictet) (12.8°C, 6.1–19.4°C). Few eggs hatched at 22.r°C.
3. The relationship between incubation period (d days) and water temperature (T°C) was given by: d=1219/T1.368 for S. torrentium , d=253/T0.459 for C. tripunctata . Both equations successfully predicted incubation periods for eggs placed in a stream. The period over which eggs hatched was much longer for C. tripunctata than for S. torrentium at all temperatures.
4. The shorter incubation period (at r>5.6°C) and shorter hatching period for S. torrentium ensure that larvae of this species are already growing when eggs of C. tripunctata start to hatch, but the prolonged hatching period of the latter species ensures a long period of larval recruitment to the population. These differences in egg hatching may reduce competition between the two closely-related species.  相似文献   

18.
Mass isolation of pure mitotic apparatuses (MAs) from sea urchin eggs was achieved using a glycerol/Mg2+/Triton X-100 isolation medium. The Mg ions stabilized the fibrous structures of the spindle and asters, while Triton X-100 favored dispersion of cell membranes. The MAs were stable for at least 1 day at 20 °C as indicated by phase contrast microscopy. The MAs also showed stable birefringence and solubility properties over a period of several hours. Only centrospheres remained intact in 0.4 M KCl-containing isolation medium. The 0.4 M KCl extract contained tubulin as one of its major components. Transfer of isolated MAs to an Mg-free medium caused the otherwise stable MA birefringence to decay upon addition of sulfhydryl-blocking reagents or Ca ions that depolymerize MA microtubules. Furthermore, when Mg ions were omitted from the isolation medium, only unstable MAs were obtained. This method seems to be of great advantage in the preparation of pure MAs in large quantity.  相似文献   

19.
  1. At temperature levels from 10 to 25°C animals from resting eggs produce subitaneous eggs independent on temperature. In contrast animals from subitaneous eggs produce subitaneous eggs dependent on temperature. At a high rate subitaneous eggs are only formed at temperature levels above 20°C.
  2. Below 10°C no development occurs in the juveniles. At temperatures of 30/22°C (24.7°C) the first subitaneous eggs are formed after 6–9 days, at 14/9°C (10.7°C) they are formed after 34 days. At different temperature levels the developmental rate of the young is from 10.5 to 42 days. One generation extends over 16.5 (30/22°C) to 75 days (14/9°C). The average egg production is 10–20 subitaneous eggs or 30–60 resting eggs. The maximum egg production of one individual is 50 subitaneous eggs or 84 resting eggs. 50% of the animals have just formed resting eggs, before the juveniles are hatched. Resting eggs in the first egg-batch are formed 6–20 days later than subitaneous eggs. The duration of life is between 65 (30/22°C) and 140 days (19/13°C).
  3. Young worms in resting eggs have a dormance period of at least 15–30 days.
At room temperatures (20°C) no juvenile in resting eggs hatches from water. By combining room and refrigerator (3.5°C) temperatures the hatching rate increases to a maximum of 85%. To reach a hatching rate of 50–65% the influence of low temperatures must be at least 30 days. At room temperatures 60% of the young in resting eggs hatch from mud covered with water. Combining high and low temperatures the hatching success is between 67 and 81%, where the highest percentage of the young may hatch at room temperature. Up to 90 days low temperatures cause a maximum hatching rate of 79%. It decreases to approximately 30% after 180 days. At high temperatures resting eggs preserved in 100% moist mud, survive for two months. By adding a period of low temperatures the hatching rate increases to a maximum of 52%. Low temperatures are survived for more than 6 months. Up to 30 days preservation at 3.5°C causes a maximum hatching rate of 61%, up to 12o days it decreases to 30%. At room temperature the young in resting eggs are not resistant against air-dried mud (30–40% rel. air moisture). Combining high and low temperatures air-dried mud is endured 1 month (hatching rate 5–14%). Preservation of 30–120 days at 3.5°C and 70% rel. air moisture result in a hatching rate of 43–61%. li]4. In the open air in Middle-Europe there occur 5–6 generations of M. ehrenbergii per life-cycle. The first generation hatches from resting eggs in May, where the production of subitaneous eggs is independent on temperature. All other generations up to October hatch from subitaneous eggs. The egg-production of those worms is dependent on environmental factors. In summer subitaneous egg production prevails, in autumn resting egg production. The abundance during the life-cycle is dependent on the number of animals which produce subitaneous eggs. Resting eggs are predestinated to endure periods of dryness and cold. The life-cycles of the species M. lingua and M. productum are different from those of M. ehrenbergii in length and in the number of generations. In both species 7 generations occur over 8 to 8.5 respectively 5.5 months. M. nigrirostrum only forms resting eggs. The life-cycle consists of one generation from February/March to May/June.  相似文献   

20.
The speed of development and the developmental mortality of Tribolium castaneum were studied over a series of temperatures between 15 and 40°C. at 10, 30, 70 and 90% R.H. using wheatfeed as food.
Eggs did not hatch at any humidity at 17·5°C. or less, nor at 10% R.H. at 40°C. At all other conditions about 80% of eggs hatched. Humidity does not affect the duration of the egg period. The effect of temperature on the egg period is shown graphically, the shortest period occurring at 37·5°C.
Larvae fail to develop into normal adults at 20°C. and also fail at 90 and 30% R.H. at 40°C. At 20°C. and 70% R.H. pupae may be formed but they do not become normal adults. The rate of larval development is affected by both temperature and humidity. Development is quickest at the highest humidity used at any temperature and at 35°C. for each humidity. Larval mortality is less than 20% except at 40°C. and at combinations of low humidity and low temperature. The results correspond with published results using similar foodstuffs.
Groundnuts were used as a food in a series of experiments at 30°C. and another series at 70% R.H. Larvae on this food were much more susceptible to the effects of humidity and were unable to develop at 20 or 40°C. at 70% R.H. In all conditions development was slower and mortality higher than on wheatfeed.
The pupal period is not affected by humidity and is shortest at 37·5°C.
The conditions in which Park found that cultures of T. castaneum became extinct, 24°C. and 30% R.H., lie inside the zone in which development is possible. The possible causes for this discrepancy are discussed.  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