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21.
Khattiya Akarasanon Praneet Damrongphol & Wandee Poolsanguan 《Aquaculture Research》2004,35(15):1415-1420
Long‐term cryopreservation of the giant freshwater prawn, Macrobrachium rosenbergii, spermatophores using glycerol (Gly) and ethylene glycol (EG) as cryoprotective agents (CPAs) was studied. The tolerance of sperm to cryopreservation was evaluated on the basis of sperm survival and fertilizing ability. The survival of the sperm was determined by trypan blue staining, while the fertilizing ability was assessed from artificial insemination of the cryopreserved spermatophores. The rates of embryo survival on day 5 after spawning and of spermatophores capable of producing embryos survived to hatching were determined. Storage of spermatophores at ?20°C without CPA for a short period of up of 1–5 days decreased the sperm survival significantly and did not preserve fertilizing ability. Preservation at ?20°C in the presence of 10% or 20% Gly or of 10% or 20% EG offered a simple and efficient short‐term storage up to 10 days. For a long‐term storage, cryopreservation in the presence of 20% EG at ?196°C was more efficient than at ?20°C. High sperm survival rates and high fertilizing ability were recorded from those cryopreserved at ?196°C for up to 150 days. High sperm survival rates with moderate levels of fertilizing ability were obtained from those cryopreserved at ?20°C for not more than 30 days. The results indicate that preservation at ?196°C with 20% EG is a suitable procedure for long‐term storage of the giant freshwater prawn spermatophores. 相似文献
22.
The effects of four cryoprotectants (methanol, MeOH; dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; and ethylene glycol, EG), three extenders (calcium‐free Hanks' balanced salt solution, C‐F HBSS, Hanks' balanced salt solution, HBSS and sodium chloride, NaCl) and two different freezing procedures (one‐ and two‐step) on the cryopreservation of striped catfish (Pangasius hypophthalmus (Sauvage)) sperm were investigated. Sperm were frozen using a controlled‐rate freezer in 250 μL straws and stored for 2 weeks in a liquid nitrogen (LN2) container. They were then airthawed at room temperature, and fertilization, motility and viability were assessed. The highest fertilization rate of 41% (81% of control) was achieved with the combination of 12% DMSO and 0.9% NaCl using a one‐step freezing procedure (10°C min?1). Also, DMA resulted in a higher fertilization rate (30% or 51% of the control) than MeOH (18% or 38% of the control) or EG (8% or 12% of the control). In addition, the three extenders used did not affect fertilization rates after cryopreservation with each cryoprotectant. There were no significant differences among the three cryoprotectant concentrations and between the one‐ and two‐step freezing procedures. However, fertilization rates of cryopreserved sperm were significantly lower than the controls (P<0.05). The results of this study indicate that high fertilization rates of striped catfish eggs can be achieved using cryopreserved sperm when frozen at 10°C min?1 in DMSO or DMA with either 0.9% NaCl or C‐F HBSS. 相似文献
23.
Short-term storage and cryopreservation of Urechis unicinctus (Echiura: Urechidae) sperm 总被引:2,自引:0,他引:2
Kyoung Ho Kang Ming Yu Shao Kang Hee Kho & Zhi Feng Zhang 《Aquaculture Research》2004,35(13):1195-1201
In the present study, attempts were made to preserve Urechis unicinctus sperm at 4°C. Cryopreservation procedures were optimized for various cryoprotectants and freezing rates, equilibration times and dilution ratios. During short‐term storage, the motility of undiluted sperm was extended for 6 days of cold storage,and in 70% and 100% artificial seawater only persisted for 2 and 4 days respectively. The survival rate of undiluted sperm was maintained at a high level accordingly. After cryopreservation, the highest motility and survival rate (41.5±2.2%) were obtained in 15% dimethyl sulphoxide (Me2SO) using a freezing rate of 30°C min?1. After thawing the sperm cryopreserved in glycerol lost almost all motility. The motility and survival rate of post‐thawing sperm did not show significant differences after 8 and 15 min equilibration using 15% Me2SO as cryoprotectant; the values were significantly higher than those of 2 min equilibration. Comparisons of motility and survival rate between treatments pooled by dilution ratio showed that the effect of 1:1 ratio (sperm volume to cryoprotectant volume) was best. There was no difference between 1:3 and 1:5, and other ratioswere significantly worse. 相似文献
24.
25.
Oxytocin (OXT) contained in boar semen is known to produce uterine contraction; therefore, we hypothesized that the co‐injection of OXT with sperm would improve artificial insemination (AI) using liquid or frozen‐thawed boar sperm. We initially examined whether OXT added to semen extender improved sperm transport to the oviduct. Although the addition of OXT did not affect the fresh or frozen‐thawed sperm motility or acrosomal integrity, it significantly increased the number of sperm in the oviduct at 6 h after AI injection with OXT, as compared with the control (P < 0.05). Moreover, some sperm were observed in the sperm reservoir of the isthmus in the OXT treatment group, whereas few sperm were observed in the control. When OXT was added to the semen extender immediately prior to AI, the conception rates were significantly higher in both fresh semen and frozen‐thawed semen than in the control group (P < 0.05: liquid, 87.5% vs. 70.5%; frozen‐thawed, 89.8% vs. 75.0%). From these results, we concluded that the addition of OXT to the semen extender assisted in sperm transportation from the uterus to the oviduct, which resulted in improved reproductive performance. 相似文献
26.
J. F. Asturiano L. Pérez D. L. Garzón F. Marco-Jiménez D. S. Peñaranda J. S. Vicente M. Jover 《Fish physiology and biochemistry》2004,30(3-4):283-293
The high sperm density, together with the short spermatozoa swimming time, makes European eel sperm manipulation and assessment
for quality difficult. Two diluting media (K15 and K30) previously designed for Japanese eel sperm were tested. After 24 h,
European eel sperm showed significant reduction in the percentage of motile spermatozoa after activation and different motility
parameters (VAP, angular velocity; VCL, curvilinear velocity; VSL, straight line velocity; BCF, beating cross frequency),
concluding that these media are not suitable to preserve the sperm of this species. After a hormonal treatment to induce spermiation,
sperm volume, density and motility were recorded at weekly samplings. The variation of the osmolality (325–330 mOsm kg−1), pH (8.4–8.6) and the ionic composition (concentration of Na+, K+, Mg2+ and Ca2+) of the seminal plasma were registered. Physio-chemical results were related with sperm quality throughout the treatment,
to determine which must be the suitable characteristics of one extender for the sperm of this species, and to find the best
conditions to obtain suitable cryopreservation media for European eel sperm. K+ concentration increased, while Ca2+ and Mg2+ concentrations showed a progressive reduction in correlation with the sperm quality improvement. Na+ showed a decreasing, but not significant tendency. P1 and P2 freezing media were designed considering the physio-chemical
parameters as well as the ionic composition shown by the best quality sperm samples, and then compared with the previously
described solutions, TNK and K30. Sperm quality was determined, checking the percentage of motile spermatozoa and motility
parameters using computer-assisted sperm analysis (CASA) software. Samples were frozen after dilution (1:5, 1:20, 1:100) in
different freezing media supplemented with 10% dimethyl sulfoxide (DMSO). After thawing, samples frozen with low dilution
ratio (1:5) in TNK and P1 media showed higher, although not significant, spermatozoa survival (35.5 ± 14.5 and 36.6 ± 6.7%).
The addition of l-α-phosphatidylcholine to the media seems to have a positive effect, as reported in the Japanese eel. 相似文献
27.
在室温、4℃、-20℃和液氮(-196℃)下贮藏小黑麦的花粉,研究不同温度对新小黑麦1号、3号、4号和5号花粉活力的影响,结果表明:室温条件下保存3 h,小黑麦花粉活力均在80%左右,3~4 h后花粉活力降低很快,7~8h后花粉活力几乎全无。4℃保存可适当延长花粉活力,保存8 h后,新小黑麦1号、5号花粉的活力降低到60%,而3号和4号降低为80%左右。48 h左右花粉活力降至0%。与4℃相比,-20℃保存下不同品种小黑麦花粉活力降低更快,4 h后花粉活力降低到50%~65%,16 h后花粉活力接近0%。液氮超低温冷冻保存下小黑麦花粉活力相对稳定,保存10天后活力仍然很高,其活力相对保持率在92%左右,理论上可以长时间保存花粉。 相似文献
28.
Kyoung Ho Kang Kang Hee Kho Zong Tao Chen Jae Min Kim Young Hun Kim & Zhi Feng Zhang 《Aquaculture Research》2004,35(15):1429-1433
The present study examined the possibility of long‐term storage, by cryopreservation in liquid nitrogen, of the sperm of filefish (Thamnaconus septentrionalis). Changes in motility, survival rate, ultrastructure and fertilization rate of the sperm after freezing and thawing were tested. For selection of the immobilizing solution, artificial seawater (ASW) of 250, 350 and 450 mOsmol kg?1 were tested. Sperm motility was significantly inhibited in 350 mOsmol kg?1 ASW, and restored entirely after 100% ASW (1200 mOsmol kg?1) was added. Two cryoprotectants, dimethyl sulphoxide and glycerol, were employed. The sperm was diluted at the ratio of 1:6 with the extenders, and frozen at a freezing rate of ?40°C min?1 to ?100°C after equilibration for 10 min at room temperature, followed by plunging into liquid nitrogen. The highest post‐thawed sperm motility and survival rate were obtained with 5% glycerol. Afterwards, the effect of different freezing rates was examined using 5% glycerol as a cryoprotectant, and the rate of ?30°C min?1 to ?100°C showed the best result. 相似文献
29.
ZHA Xing-qin XIAO Jing CHENG Wen-min HUO Jin-long WANG Shu-yan WANG Pei PAN Wei-rong ZENG Yang-zhi 《中国畜牧兽医》2017,44(1):167-172
In this study, different concentrations of glycerol were added to semen cryoprotective extenders of Banna Mini-pig inbred line (BMI), different thawing solutions and thawing methods were studied. After thawing, the motility, the rates of plasma integrity and acrosome integrity were compared to optimize the cryopreservation method of BMI semen. The results showed that the motility, the rates of plasma integrity and acrosome integrity were significantly higher in 2% and 3% glycerol groups than those in 1%, 4% and 5% glycerol groups (P<0.05);The motility, the rates of plasma integrity and acrosome integrity were significantly higher in thawing solution Ⅰ than those in thawing solution Ⅱ, Ⅲ and Ⅳ (P<0.05); Through comparing the motility, the rates of plasma integrity and acrosome integrity, the thawing effect in 40℃ 6 s and 50℃ 6 s groups were better than that in 38℃ 30 s (P<0.05). In conclusion, the optimal semen cryoprotective method for BMI was 2% or 3% glycerol as cryoprotectant, using thawing solution Ⅰ to thaw at 40℃ 6 s or 50℃ 6 s. 相似文献
30.
系统地阐述了组织培养技术在甜菜快速繁殖、创造新种质以及种质保存等方面的应用研究进展。同时对甜菜组织培养未来的发展方向进行了展望。 相似文献