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Accurate identification of bovine parainfluenza type 3 virus in bovine respiratory disease requires dependable, sensitive, and specific techniques for detection in affected animals. Immunohistochemical testing can be a rapid and reliable means of demonstration of virus in tissues from suspect cases; however, this procedure is dependent upon the quality of the antisera directed against the viral antigens. The production of rabbit polyclonal and murine monoclonal antibodies directed against bovine parainfluenza type 3 virus and techniques for their use in fresh-frozen and formalin-fixed paraffin-embedded tissues in immunofluorescence and immunoperoxidase-based immunohistochemical tests are described.  相似文献   
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Sixty-eight cases of suppurative, ulcerative endometritis associated with Bovine Herpesvirus-4 (BHV-4) in postparturient dairy cows (62 Holsteins and 6 Jerseys, mean age 4.2 years) were confirmed by a combination of histopathology, fluorescent antibody assays, electron microscopic evaluation of uterus, and polymerase chain reaction (PCR). All cases occurred in the 3- to 28-day postpartum period, and histologic lesions among various cows were consistent when compared with postpartum interval. The endometrial lining epithelium was necrotic and ulcerated from 3 to 7 days postpartum, with only mild inflammation in the lamina propria and submucosa. From 1 to 4 weeks postpartum, the ulcers were confluent to diffuse. Epithelium was replaced by fibrinonecrotic, suppurative mats, resulting in severe bacterial pyometra by day 24. Seroprevalence to BHV-4 in one dairy with a history of 18 mortality cases was 36% (107 of 296). In a random sample of 8 cows from this herd, none had serologic titers in blood sampled 2 weeks prepartum, but 3 of 8 seroconverted with significant titers of 1:8 to 1:16 at 2 weeks postpartum. By 10 weeks postpartum, all 8 cows returned to negative serologic status. Two of 6 cats from the premises also had positive titers. Random serum samples taken from 480 dairy cattle at sale barns indicated 76 (16%) were positive by serum neutralization. Clinical signs, postparturient timing, and histologic lesions were very similar to those previously reported in Belgium with BHV-4. But sequence analysis of PCR products of the glycoprotein B region of 4 separate field isolates of endometriotropic BHV-4 suggests these field isolates were more closely related to the North American nonvirulent strain DN-599 than to the endometriotropic European strain V.  相似文献   
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OBJECTIVE: To determine whether antemortem core needle biopsy and fine-needle aspiration of enlarged peripheral lymph nodes could be used to distinguish between inflammation and lymphosarcoma in cattle. DESIGN: Prospective study. ANIMALS: 25 cattle with enlarged peripheral lymph nodes. PROCEDURES: Antemortem biopsies of the selected lymph nodes were performed with an 18-gauge, 12-cm core needle biopsy instrument. Fine-needle aspirates were performed with a 20-gauge, 4-cm needle. Specimens were analyzed by pathologists who were unaware of clinical findings and final necropsy findings, and specimens were categorized as reactive, neoplastic, or nondiagnostic for comparison with necropsy results. RESULTS: Sensitivity and specificity of core needle biopsy ranged from 38% to 67% and from 80% to 25%, respectively. Sensitivity of fine-needle aspiration ranged from 41% to 53%, and specificity was 100%. Predictive values for positive test results ranged from 77% to 89% for core needle biopsy and were 100% for fine-needle aspiration. Predictive values for negative test results were low for both core needle biopsy and fine-needle aspiration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that core needle biopsy and fine-needle aspiration can aid in the antemortem diagnosis of bovine enzootic lymphosarcoma. Results of fine-needle aspiration of enlarged peripheral lymph nodes were more specific and more predictive for a positive test result than were results of core needle biopsy.  相似文献   
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M. J. Allison  H. Bain 《Euphytica》1986,35(2):345-351
Summary A reversed-phase high performance liquid chromatography (RP-HPLC) system was used to separate the storage proteins (hordeins) extracted from European barley cultivars. From a total of 38 barleys tested, 26 types of hordein patterns could be distinguished after RP-HPLC. This appears to be a marked improvement in resolution over that achieved in a similar survey of European barley cultivars using SDS polyacrylamide electrophoresis (32 hordein patterns resolved by SDS PAGE from a total of 160 spring and winter barleys tested).Different hordein patterns were resolved by RP-HPLC within each of two groups of barley previously classified by SDS PAGE as indistinguishable within groups (three distinct patterns identified in a total of five cultivars tested from group 1A and five patterns observed among eight cultivars from group 3B). Thus RP-HPLC achieves a higher resolution than undirectional electrophoresis and promises to be a valuable aid in the identification of European barley cultivars.  相似文献   
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We describe here the results of a study conducted to evaluate a terminal restriction fragment length polymorphism (TRFLP) approach targeting rRNA genes for determination of ectomycorrhizal (ECM) communities colonizing the roots of loblolly pine (Pinus taeda L.). Root tips separated from soil cores were classified according to morphological characteristics and DNA was then extracted from each group of morphotyped tips. Labeled primers were used to generate terminal restriction fragments (TRF) for molecular fingerprinting of root colonizing fungi and to determine how well TRFLP could be used to discriminate between ectomycorrhizal types. Morphotypes generally correlated well with specific TRFs and sequence analysis confirmed that TRFs could be used to discriminate among fungal types. Sequence analysis indicated that important ECM fungi including Russulaceae, Thelephorales, and Tricholomataceae could be fingerprinted with TRFLP. In addition, a fixed proportion of the DNA extracted from each morphotype from the same core was used in a pooling experiment used to assess whether previously identified fungal species types could be distinguished from one another within reconstructed communities. Since some morphotypes share TRFs, dual analysis of ITS1 and ITS2 was necessary for accurate fingerprinting of fungal types. Approximately, 5.0±0.3 phylotypes were detected per core with TRFLP-corrected morphotyping as compared to 4.0±0.4 phylotypes using TRFLP on pooled community samples. TRFLP made on experimental sporocarp communities suggested that reduced ECM richness with TRFLP may be partly caused by differences in the amount of DNA available for PCR and primer bias. Nonetheless, TRFLP on pooled morphotypes accounted for more than 93% of colonized root tips. The method can be used to facilitate analysis of mycorrhizal communities using root tips collected from soil cores.  相似文献   
39.
OBJECTIVE: To evaluate responses to self-administered brief questions regarding consumption of vegetables and fruit by comparison with blood levels of serum carotenoids and red-cell folate. DESIGN: A cross-sectional study in which participants reported their usual intake of fruit and vegetables in servings per day, and serum levels of five carotenoids (alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin and lycopene) and red-cell folate were measured. Serum carotenoid levels were determined by high-performance liquid chromatography, and red-cell folate by an automated immunoassay system. SETTINGS AND SUBJECTS: Between October and December 2000, a sample of 1598 adults aged 25 years and over, from six randomly selected urban centres in Queensland, Australia, were examined as part of a national study conducted to determine the prevalence of diabetes and associated cardiovascular risk factors. RESULTS: Statistically significant (P<0.01) associations with vegetable and fruit intake (categorised into groups: /=4 servings per day) were observed for alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin and red-cell folate. The mean level of these carotenoids and of red-cell folate increased with increasing frequency of reported servings of vegetables and fruit, both before and after adjusting for potential confounding factors. A significant association with lycopene was observed only for vegetable intake before adjusting for confounders. CONCLUSIONS: These data indicate that brief questions may be a simple and valuable tool for monitoring vegetable and fruit intake in this population.  相似文献   
40.
Simultaneously assessing shifts in microbial community composition along landscape and depth gradients allows us to decouple correlations among environmental variables, thus revealing underlying controls on microbial community composition. We examined how soil microbial community composition changed with depth and along a successional gradient of native prairie restoration. We predicted that carbon would be the primary control on both microbial biomass and community composition, and that deeper, low-carbon soils would be more similar to low-carbon agricultural soils than to high carbon remnant prairie soils. Soil microbial community composition was characterized using phospholipid fatty acid (PLFA) analysis, and explicitly linked to environmental data using structural equations modeling (SEM). We found that total microbial biomass declined strongly with depth, and increased with restoration age, and that changes in microbial biomass were largely attributable to changes in soil C and/or N concentrations, together with both direct and indirect impacts of root biomass and magnesium. Community composition also shifted with depth and age: the relative abundance of sulfate-reducing bacteria increased with both depth and restoration age, while gram-negative bacteria declined with depth and age. In contrast to prediction, deeper, low-C soils were more similar to high-C remnant prairie soils than to low-C agricultural soils, suggesting that carbon is not the primary control on soil microbial community composition. Instead, the effects of depth and restoration age on microbial community composition were mediated via changes in available phosphorus, exchangeable calcium, and soil water, together with a large undetermined effect of depth. Only by examining soil microbial community composition shifts across sites and down the soil column simultaneously were we able to tease apart the impact of these correlates environmental variables.  相似文献   
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