Comparison of the TBARS Assay and BODIPY C11 Probes for Assessing Lipid Peroxidation in Red Deer Spermatozoa     

ÁE Domínguez‐Rebolledo  F Martínez‐Pastor  MR Fernández‐Santos  E Del Olmo  A Bisbal  JL Ros‐Santaella  JJ Garde 《Reproduction in domestic animals》2010,45(6):e360-e368

Several methods are used to measure lipid peroxidation (LPO) in spermatozoa. The objective of this study was comparing the thiobarbituric acid reactive species (TBARS) method and the BODIPY 581/591 C₁₁ (B581) and BODIPY 665/676 C₁₁ (B665) fluorescent probes to measure induced peroxidative damage in thawed epididymal spermatozoa from Iberian red deer. Samples from three males were thawed, pooled, diluted in PBS, incubated at room temperature and assessed at 0, 3, 6 and 24 h under different experimental conditions: Control, hydrogen peroxide (H₂O₂) 0.1 mm or 1 mm , or tert‐butyl hydroperoxide (TBH) 0.1 mm or 1 mm . LPO was assessed by the TBARS assay [malondialdehyde (MDA) detection] and by the fluorescence probes B581 and B665 (microplate fluorimeter and flow cytometry). Increasing MDA levels were only detectable at 1 mm of TBH or H₂O₂. Both fluorescence probes, measured with fluorometer, detected significant increases of LPO with time in all treatments, except Control. Flow cytometry allowed for higher sensitivity, with both probes showing a significant linear relationship of increasing LPO with time for all oxidizing treatments (p < 0.001). All methods showed a good agreement, except TBARS, and flow cytometry showed the highest repeatability. Our results show that both B581 and B665 might be used for LPO analysis in Iberian red deer epididymal spermatozoa, together with fluorometry or flow cytometry. Yet, the TBARS method offered comparatively limited sensitivity, and further research must determine the source of that limitation.  相似文献   

Horizontal transmission of pseudorabies virus in cattle     

R A Crandell  G M Mesfin  R E Mock 《American journal of veterinary research》1982,43(2):326-328

Pseudorabies virus (PRV) was not transmitted horizontally from 3 PRV-infected calves to 2 contact control calves during 4 days of comingling in experiment 1. Although these contact control calves developed clinical signs of pseudorabies when infected intranasally with PRV in experiment 2, they did not transmit PRV to a second pair of contact control calves. However, 1 of 2 pigs comingled with these 4 calves seroconverted. During both experiments, moderate amounts (10(2) to 10(5) TCID50) of PRV were present in the nasal secretions of the infected calves during the contact periods. All infected calves traumatized their nares or periorbital tissue. Infected calves developed a nonsuppurative meningoencephalitis mainly involving the brain stem. Four of the 5 infected calves had nonsuppurative ganglioneuritis and acute lymphoid necrosis of germinal centers. Virus could not be recovered from nasal and tonsillar swab samples from contact-control calves and pigs.  相似文献   

Effects of Cryopreservation on Bull Spermatozoa Distribution in Morphometrically Distinct Subpopulations     

J Rubio-Guillén  D González  JJ Garde  MC Esteso  MR Fernández-Santos  JE Rodríguez-Gíl  N Madrid-Bury  A Quintero-Moreno 《Reproduction in domestic animals》2007,42(4):354-357

Assisted sperm morphometry analysis (ASMA) was used in this study to determine the effects of cryopreservation on bull spermatozoa distribution in morphometrically distinct subpopulations. Ejaculates were collected from five bulls and were divided. One portion was diluted at 30 degrees C in a skim milk-egg yolk medium, containing glycerol. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours. After thawing, sperm smears were prepared as described above. All slides were air dried and stained with Hemacolor. The sperm-head dimensions for a minimum of 200 sperm heads were analysed from each sample by means of the Sperm-Class Analyser (SCA), and the mean measurements recorded. Our results showed that applying the ASMA technology and multivariate cluster analyses, it was possible to determine that three separate subpopulations of spermatozoa with different morphometric characteristics coexist in bull ejaculates (large, average and small spermatozoa). The mean values of each sperm head dimension among the three subpopulations of spermatozoa were significantly different (p < 0.001). Besides, there were significant (p < 0.001) differences in the distribution of these three sperm subpopulations between fresh and thawed samples. Thus, the percentage of representation of the subpopulation that includes those spermatozoa whose dimensions are the biggest, decreased from 52.06% in extended fresh samples to 15.51% in the thawed ones. Contrarily, the percent of representation of the subpopulation containing the smallest spermatozoa, increased from 8.70% in extended fresh samples to 34.04% in the thawed ones. In conclusion, the present study confirms the heterogeneity of sperm head dimensions in bull semen, heterogeneity that vary through the cryopreservation procedure.  相似文献   

Effects of recurrent selection for grain yield on plant and ear traits of five maize populations     

Theodore M. Crosbie  James J. Mock 《Euphytica》1980,29(1):57-64

Summary Maize (Zea mays L.) breeders are interested in the effects of recurrent selection for grain yield on other traits. Changes in plant traits could alter agronomic acceptability of the populations under selection, and observed improvements in grain yield could be explained by changes in ear traits. We evaluated changes in combining ability for plant and ear traits of BS10(FR), BS11(FR), BSSS(R), BSCB1(R), and Lancaster Surecrop associated with recurrent selection for grain yield.Recurrent selection procedures generally did not change plant and ear heights or date of silking of testcrosses of populations or of the population crosses, BS10(FR)×BSS11(FR) and BSSS(R)×BSCB1(R). Grain yield improvements, however, were associated with increases in ear-sink size.Journal Paper No. J 9517 of the Iowa Agriculture and Home Economics Exp. Stn., Ames, Iowa. Project 2152.  相似文献   

An ideotype of maize   总被引：9，自引：0，他引：9  

J. J. Mock  R. B. Pearce 《Euphytica》1975,24(3):613-623

Summary We have proposed breeding a maize (Zea mays L.) ideotype that will maximally utilize an optimum production environment. This environment should include: a) adequate moisture; b) favorable temperatures throughout the growing season; c) adequate fertility; d) high plant densities; e) narrow row spacings; and f) early planting dates. The maize ideotype that should produce optimally when grown in such an environment would be characterized by: a) stiff, vertically-oriented leaves above the ear (leaves below the ear should be horizontally oriented); b) maximum photosynthetic efficiency; c) efficient conversion of photosynthate to grain; d) short interval between pollen shed and silkemergence; e) ear-shoot prolificacy; f) small tassel size; g) photoperiod insensitivity; h) cold-tolerance in germinating seeds and young seedlings (for genotypes grown in areas where early-planting would require planting in cold, wet soils); i) as long a grain-filling period as practically possible; and j) slow leaf senescence.Journal Paper No. J-8108 of the Iowa Agriculture and Home Economics Experiment Station. Ames, IA 50010. Projects 1836 and 1990.  相似文献   

Development of a multiplex TaqMan real-time RT-PCR assay for simultaneous detection of Asian prunus viruses, plum bark necrosis stem pitting associated virus, and peach latent mosaic viroid     

Liming Lin  Ruhui Li  Margarita Bateman  Raymond Mock  Gary Kinard 《European journal of plant pathology / European Foundation for Plant Pathology》2013,137(4):797-804

Asian prunus viruses (APV 1, APV 2 and APV 3), Plum bark necrosis stem pitting associated virus (PBNSPaV) and Peach latent mosaic viroid (PLMVd) are pathogens that infect Prunus species. A single-tube multiplex, TaqMan real-time RT-PCR assay was developed for the simultaneous detection and identification of these pathogens. The protocol includes amplification and detection of a fluorogenic cytochrome oxidase gene (COX) as an internal control. The results of the multiplex TaqMan RT-PCR assay correlated with those from conventional RT-PCR, with a 10-fold increase in sensitivity in the multiplex real-time format. The efficiency and accuracy of the assay was evaluated by testing stone fruit trees from positive control collections and several orchard locations. Several mixed infections of target pathogens were detected in peach orchard samples. This assay is simple, rapid and cost-effective and can be used by quarantine and certification programs where numerous stone fruit trees need to be tested for these pathogens.  相似文献   

Photopolymerization and Mass-Independent Sulfur Isotope Fractionations in Carbon Disulfide     

JJ Colman  X Xu  MH Thiemens  WC Trogler 《Science (New York, N.Y.)》1996,273(5276):774-776

Irradiation of gaseous carbon disulfide [CS2(g)] at 313 nanometers produces a dark brown aerosol of (CS2)x. Its thermal decomposition products include disulfur (S2), carbon monosulfide (CS), and (CS)x. The photopolymerization process is accompanied by a large mass-independent isotopic fractionation of sulfur (a 5 to 10 per mil sulfur-33 excess and a 61 to 84 per mil sulfur-36 deficit). Excess sulfur-33 has been observed in several classes of meteorites. Photochemical production of (CS2)x may be important in the origin and evolution of cosmochemical environments such as the presolar nebula, meteorites, asteroids, and planetary atmospheres.  相似文献   

DNA Status on Thawed Semen from Fighting Bull: A Comparison Between the SCD and the SCSA Tests     

F Martínez-Pastor  M del Rocío Fernández-Santos  ÁE Domínguez-Rebolledo  MC Esteso  JJ Garde  ; Biology of Reproduction Group 《Reproduction in domestic animals》2009,44(3):424-431

The assessment of sperm chromatin status is compulsory in a complete spermiogram. Here we applied the sperm chromatin structure assay (SCSA) and the sperm chromatin dispersion (SCD) test to assess the chromatin status of three fighting bulls. Cryopreserved semen (two straws/bull) were analysed by duplicate after thawing and after 6 h at 37°C with and without oxidative stress (1 m m FE²⁺). Results (SCD: percentage of spermatozoa with halo; SCSA: SD-DFI, %DFI and HDS) were analysed for differences between bulls and treatments, sensitivity and specificity (receiver operating characteristic curves) and repeatability (repeatability coefficients as 2SD of duplicate differences).%DFI for the three bulls was below 2% at 0 h, indicating no risk for fertility according to previous reports. It increased slightly for two of the bulls after FE²⁺ treatment (%DFI < 5%) and more pronouncedly for the other bull (C, %DFI∼10%), which merits further investigation. SCD rendered higher percentage of halos for bull C, but could not discriminate between samples with and without oxidizing treatment (AUC: 0.52). SCSA (%DFI) showed a high discriminating ability between treatments (AUC: 0.96). The repeatability coefficient was also higher for SCD (5.9) than for %DFI (1.8), indicating lower repeatability for SCD. Overall, %DFI might be the most useful parameter for assessing sperm chromatin on fighting bull. SCD might yield different information than SCSA, hence further research is warranted.  相似文献