大熊猫冷冻精液解冻速度和解冻液中添加Pentyoxyfilline对其解冻后活力的影响

建立大熊猫的精子库,进行远距离圈养大熊猫种群间的人工授精和遗传物质的转运,维持遗传多样性,是目前大熊猫遗传管理的优先方法。要成为最有效的工具,精子库保存的精子解冻后的活力必须很好。本文对大熊猫冷冻精液的解冻速度和解冻液中添加化学激活剂Pentyoxyfilline(PF)后的精子活力进行了试验。试验用的精液采自11只成年大熊猫,精液冷冻速度为每分钟-40℃～-100℃。试验Ⅰ:将冷冻精液放入3种不同温度的水浴中解冻:(1)22℃(慢速解冻);(2)37℃(中速解冻)(3)50℃(快速解冻)。将冷冻前精子活力(78 1±2 9%)和解冻后的平均精子活力进行比较,快速解冻后的精子活力(57 5±5 4%)显著地降低(P<0 05),而中速解冻的精子活力(67 5±3 1%)和慢速解冻的精子活力(73 33±2 1%)与冷冻前的活力接近。试验Ⅱ:使用中速解冻方法解冻精液后,分别加入最终浓度为0mM、1mM、5mM和10mM的PF,然后分别保温15min和24h。在PF(0mM、1mM、5mM和10mM)中分别孵育15min的解冻精子活力,运动状态,活率和顶体正常率在试验期的90min内都很相似(P>0 05)。在1mMPF中孵育24h的精子活力没有变化(P>0 05)。在5mM和10mMPF中孵育过的精子活力(5mM:24 0±4 7%;10mM19 5±3 6%)比没有加PF的对照组的精子活力(38 3±5 2%)显著地低(P<0 05)。而且,在10mM     

Influence of Boar and Semen Parameters on Motility and Acrosome Integrity in Liquid Boar Semen Stored for Five Days

Ninety ejaculates from a total of 76 AI boars were extended in Beltsville Thawing Solution (BTS). Boar identity, breed, weight of the ejaculate and sperm concentration were registered. Motility and acrosome integrity were assessed after storage at 16–18°C for 6, 30, 54, 78, and 102 h. Storage time had a significant influence on both motility (p < 0.01) and acrosome integrity (p < 0.001). The Least Square Means for percentage of motility showed a small decline from 79.8% after 6 h of storage to 78.4% at 102 h. Motility at 78 and 102 h was significantly different from motility at 6 h (p < 0.05). The percentage of sperm cells with normal acrosomes declined throughout the experiment. The Least Square Means for 6, 30, 54, 78, and 102 h of storage were 93.9%, 90.6%, 88.0%, 84.8%, and 78.2%, respectively. The decrease in acrosome integrity from one storage time to the next was highly significant throughout the trial (p < 0.001). There was a significant influence of boar (p < 0.001) and sperm concentration (p < 0.01) on motility, while acrosome integrity was affected only by boar (p < 0.001). Breed of the boars and weight of the ejaculate did not influence the dependent variables.     

Relation of Chronotype to Sleep Complaints in the General Finnish Population

Individuals show variation in their preference for the daily timing of activities. In this study the authors analyzed whether chronotypes associate with sleep duration and sleep-related complaints. The authors used the National FINRISK Study 2007 Survey data on 3696 women and 3162 men, representative of the Finnish population aged 25 yrs and older, for the assessment of chronotype and self-reported sleep. Evening types experienced insomnia symptoms, had nightmares, and had used recently hypnotics significantly more often than other chronotypes among both men and women. In a multinominal logistic regression model predicting insufficient sleep, the association of eveningness with insufficient sleep was not abolished after adjustment for sex, age, and sleep duration. The prevalence of short sleepers was significantly higher in evening types among men than among women, whereas that of long sleepers was significantly higher in evening types among both men and women, as compared with the other chronotypes. These results indicate that eveningness predisposes individuals to a range of sleep complaints. (Author correspondence: ilona.merikanto@helsinki.fi)     

Allele frequencies of the major milk proteins in the Finnish Ayrshire and detection of a new K-casein variant

A total of 20990 Finnish Ayrshire cows were phenotyped for the major milk proteins by isoelectric focusing in polyacrylamide gels. The predominant alleles in the Finnish Ayrshire were α_S1-casein B (0.999), α_S2-casein A (0.991), β-casein A₁ (0.509) and α₂ (0.490), α₂₂-casein A (0.612) and β-lactoglobulin B (0.716). The K-casein E allele (0.307) was also rather common in the Finnish Ayrshire. A new K-casein variant (K-casein F) was demonstrated in two Finnish Ayrshire cows, a dam and a daughter.     

Relation of chronotype to sleep complaints in the general Finnish population

Individuals show variation in their preference for the daily timing of activities. In this study the authors analyzed whether chronotypes associate with sleep duration and sleep-related complaints. The authors used the National FINRISK Study 2007 Survey data on 3696 women and 3162 men, representative of the Finnish population aged 25 yrs and older, for the assessment of chronotype and self-reported sleep. Evening types experienced insomnia symptoms, had nightmares, and had used recently hypnotics significantly more often than other chronotypes among both men and women. In a multinominal logistic regression model predicting insufficient sleep, the association of eveningness with insufficient sleep was not abolished after adjustment for sex, age, and sleep duration. The prevalence of short sleepers was significantly higher in evening types among men than among women, whereas that of long sleepers was significantly higher in evening types among both men and women, as compared with the other chronotypes. These results indicate that eveningness predisposes individuals to a range of sleep complaints.     

Semen assessment, fertility and the selection of Hereford bulls for use in AI

Nineteen young Hereford bulls were used to study the relationship between semen characteristics and fertility in artificial insemination following 15 320 inseminations. Seven measures of sperm motility, morphological abnormalities, the release of hyaluronidase, ATP content and sperm head measurements were examined as predictors of fertility (49-day fixed-interval non-return rate). Two assessments of motility, three categories of abnormal spermatozoa, acrosomal changes and the release of hyaluronidase had predictive power. Multiple regression analysis showed that a combination of sperm motility after dilution in saline, motility after thawing and the proportion of coiled tails and proximal protoplasmic droplets provided the best prediction of fertility and allowed bulls to be ranked in order of observed non-return rate (%) with a Spearman correlation better than +0.80.     

Identification of Bovine Leucocyte Adhesion Deficiency (BLAD) Carriers in Holstein and Brown Swiss AI Bulls in Iran

BLAD is a hereditary disease in Holstein dairy cattle. The defective allele of CD18 gene, which is responsible for this disease, has recessive inheritance. The recessive homozygous form (BL/BL) is lethal and since carrier animals have viability, BLAD frequency increases by use of carrier bulls in Artificial Insemination (AI). BLAD carriers can be detected easily by means of polymerase chain reaction followed by restriction analysis of the amplicons. In this study DNA samples from Holstein (n = 30) and Brown Swiss (n = 10) bulls from Abbas Abad AI center (Khorasan state of Iran) were analysed. A 101-bp fragment from the polymorphic region of CD18 gene located on chromosome 1 was amplified by PCR. Restriction enzymes TaqI and HaeIII were used to identify genotypes. Digestion products were screened by electrophoresis on 8% non-denaturing polyacrylamide gel and visualized by ethidium bromide staining. Frequencies of BL/TL (carrier) genotypes in Holstein and Brown Swiss bulls were 3.33% and 0%, respectively. Our pedigree studies of the carrier bull in this experiment revealed that the mutation was inherited by him from Hawkeye bull (CANM 369995, BL). Although the elimination of BLAD-carrier bulls from the Holstein world would be the most efficient method to control this genetic disorder, many BLAD-carrier bulls are still listed commercially for AI, and BLAD is still occurring in Iran. Monitoring the prevalence of BLAD carriers in random selected herds may be helpful in judging the effectiveness of the BLAD-control program.     

Prion's Progress: Patterns and Rates of Molecular Evolution in Relation to Spongiform Disease

Modification of the cellular prion protein has been correlated with the acquisition of several neurodegenerative diseases, including kuru, scrapie, bovine spongiform encephalopathy (BSE), and Creutzfeldt–Jakob disease (CJD). Sequence conservation and amino acid identity are known to influence the efficacy of interspecific transmission. We analyzed patterns of interspecific genetic variation with a view toward identifying features related to disease transmission. The reconstructed gene trees and amino acid tree were compared with the species tree, and all discordances observed were related to the species barrier of disease transmission. The rates of synonymous substitution, nonsynonymous substitution, and nucleotide content were determined for the protein-coding gene. Substitutions implicated in each of the prion diseases were found to occur in regions of the protein that are least variable across all species—opposite to the pattern of variability expected from interaction with an infectious pathogen. Amino acid residues related to the species barrier form a single cluster associated with the first alpha-helical domain of the protein. Residues related to sporadic and hereditary human prion disease form two separate clusters, associated with the second and third alpha-helical domains. Taken together, these results are consistent with the view that prion diseases arise from accidents in protein folding, rather than infection with an undiscovered virus-like particle. We speculate that the differences in disease phenotype between transmissable and hereditary forms could result from interactions between different parts of the protein during propagation. Received: 18 April 1997 / Accepted: 17 October 1997     

Fermentation Rates and Ethanol Accumulation in Relation to Flooding Tolerance in Rhizomes of Monocotyledonous Species

Under anoxia an accumulation of ethanol took place in all species.Lactic fermentation was found to be of less importance in everycase. The amount of ethanol accumulated depended on the abilityof the rhizome to eliminate it from its tissues. Despite greatvariations in fermentative capacity as seen in the ADH measurementsthe wetland species did not accumulate ethanol to concentrationsgreater than 30 µmol g^–1 fresh weight. This representeda plateau above which the ethanol concentration did not riseeven with continued anaerobic incubation. No such plateau ofethanol accumulation was found in the dryland species Iris germanicawhich accumulated ethanol steadily reaching concentrations of70 µmol g^–1 fresh weight. If ethanol is toxic to higher plant tissues then the steadystate condition of low ethanol accumulation found in wetlandspecies will minimize this danger. Key words: Ethanol, Flooding, Rhizomes     

Nitrogen-Use Efficiency in Relation to Different Forms and Application Rates of Se in Lettuce Plants

The main objective of this work was to determine whether nitrogen-use efficiency (NUE) was affected by the application of different forms and dosages of selenium (Se), to ascertain the influence of this trace element in a biofortification program in lettuce plants. The parameters analyzed were biomass, NO₃ ⁻ concentration, and total reduced N as well as those defining NUE in plants: total nitrogen accumulation (TNA), nitrogen efficient ratio (NER), nitrogen-utilization efficiency (NUtE), and nitrogen-uptake efficiency (NUpE). According to our results, application of Se affected NUE in lettuce plants. With the application of selenite as well as selenate NO₃ ⁻ uptake was reduced, thus diminishing the NUpE and the foliar concentration of this anion. In addition, selenate application at a rate of 20 μM and selenite at 5 μM induced N utilization, reflected by an increase in NER and NUtE; this result coincides with augmented biomass production. Notably, our results indicate that when Se is applied at high rates, selenite is far more phytotoxic, this being associated with a higher reduction of NUE in these plants.     

Endo-[beta]-Mannanase Activity from Individual Tomato Endosperm Caps and Radicle Tips in Relation to Germination Rates

Endo-[beta]-mannanase is hypothesized to be a rate-limiting enzyme in endosperm weakening, which is a prerequisite for radicle emergence from tomato (Lycopersicon esculentum Mill.) seeds. Using a sensitive, single-seed assay, we have measured mannanase activity diffusing from excised tomato endosperm caps following treatments that alter the rate or percentage of radicle emergence. Most striking was the 100- to more than 10,000-fold range of mannanase activity detected among individual seeds of highly inbred tomato lines, which would not be detected in pooled samples. In some cases a threshold-type relationship between mannanase activity and radicle emergence was observed. However, when radicle emergence was delayed or prevented by osmoticum or abscisic acid, the initial increase in mannanase activity was unaffected or even enhanced. Partially dormant seed lots displayed a bimodal distribution of activity, with low activity apparently associated with dormant seeds in the population. Gibberellin- and abscisic acid-deficient mutant seeds exhibited a wide range of mannanase activity, consistent with their variation in hormonal sensitivity. Although the presence of mannanase activity in the endosperm cap is consistently associated with radicle emergence, it is not the sole or limiting factor under all conditions.     

The Relative Growth Rates of Three Plankton Diatoms in Relation to Underwater Radiation and Temperature

Relative growth rates of three freshwater plankton diatoms-Asterionellaformosa, Fragilaria crotonensis, and Tabellaria flocculosa var.asterionelloides-are described from cultures suspended at variousdepths and during several seasons in the lake Windermere. Seasonalvariation in rates recorded near the surface (i m. depth) isinterpreted in terms of seasonal changes in temperature anddaylength. Rates recorded for Asterionella and Fragilaria aregenerally similar, but are approximately twice the rates obtainedwith Tabellaria. Depth profiles of relative growth rates areof similar form in all species, and normally show lightsaturationnear the surface. The shape of profiles for Asterionella isin good agreement with estimates of photosynthesis integratedover the growth periods. The parallelism between photosyntheticand relative growth rates of Asterionella is further illustratedfrom laboratory experiments: an approximate interconversion,under certain conditions, is given.     

The role of the bovine growth hormone receptor and prolactin receptor genes in milk, fat and protein production in Finnish Ayrshire dairy cattle

We herein report new evidence that the QTL effect on chromosome 20 in Finnish Ayrshire can be explained by variation in two distinct genes, growth hormone receptor (GHR) and prolactin receptor (PRLR). In a previous study in Holstein-Friesian dairy cattle an F279Y polymorphism in the transmembrane domain of GHR was found to be associated with an effect on milk yield and composition. The result of our multimarker regression analysis suggests that in Finnish Ayrshire two QTL segregate on the chromosomal region including GHR and PRLR. By sequencing the coding sequences of GHR and PRLR and the sequence of three GHR promoters from the pooled samples of individuals of known QTL genotype, we identified two substitutions that were associated with milk production traits: the previously reported F-to-Y substitution in the transmembrane domain of GHR and an S-to-N substitution in the signal peptide of PRLR. The results provide strong evidence that the effect of PRLR S18N polymorphism is distinct from the GHR F279Y effect. In particular, the GHR F279Y has the highest influence on protein percentage and fat percentage while PRLR S18N markedly influences protein and fat yield. Furthermore, an interaction between the two loci is suggested.     

Changes of Exponential Growth Rates in Relation to Differentiation of Geotrichum candidum in Submerged Culture

Growth and differentiation of the imperfect fungus Geotrichum candidum were followed in submerged cultures containing a simple synthetic glucose salt medium. Uptake of glucose, ammonium and oxygen from the medium were measured during the entire growth perod. In 0.1% glucose the fungus grows with one exponential growth phase until all the glucose has been consumed. The arthrospores are formed in the stationary phase. In 0.5% glucose the growth curve has two exponential growth phases, one with a doubling time of 1.8 h and a second one with a doubling time of 4.9 h. The second exponential growth phase, which starts when less than 15% of the glucose and less than 30% of the ammonium have been consumed, is shown to be the sporulation phase. During this growth phase the oxygen saturation in the culture remained constant at about 50%.     

Aerobic Bacterial Flora of Semen and Stallion Reproductive Tract and its Relation to Fertility Under Field Conditions

This study was initiated in order to investigate the bacterial flora of the stallion genital tract by taking consecutive samples from normal stallions in regular use. The objective was to determine whether any growth of potential pathogens, particularly P. aeruginosa and K. pneumoniae, in fresh semen and urethra was associated with the presence of inflammatory cells in the semen and whether bacterial growth had any effect on sperm morphology and pregnancy results. Sixteen stallions, only used for A.I., housed at 3 different commercial stud farms, were used. A wide variety of microorganisms was found in almost all samples from fresh semen (total 115 samples). P. aeruginosa was isolated from 46/115 (40%) of the samples and from 12 of the 16 stallions. K. pneumoniae was isolated from the semen of one stallion. Samples taken from the distal urethra after ejaculation contained fewer microorganisms than samples from fresh semen. No bacteria were found in 51% of the extended semen samples. Most of the stallions had an acceptable sperm morphology, and very few of the ejaculates contained inflammatory cells. Pregnancy results among the stallions varied, but were acceptable for most of them. There was no correlation between the frequency of samples testing positive for P. aeruginosa in raw semen and pregnancy results. kw|Keywords|k]bacterial growth k]urethra k]Pseudomonas aeruginosa k]Klebsiella pneumoniae     

Assessment of Sperm Viability by Measurement of ATP,Membrane Integrity and Motility in Frozen/Thawed Bull Semen

Control of sperm quality after commercial freezing/thawing of bull semen is still restricted to the subjective assessment of sperm motility, despite its low correlation to fertility (Söderquist et al. 1991, Kjaestad et al. 1993). Although no single in vitro method has yet been designed to predict the fertilizing ability of a given semen sample, the quantitation of viable spermatozoa (with intact plasma and acrosome membranes, and metabolically active) seems to be most promising (Woelders et al. 1991). The present report describes the use of a bioluminiscence technique to determine ATP-levels and a novel supravital stain (using fluorescent dyes) to assess the amount of viable spermatozoa in frozen/thawed semen from 3 A.I. dairy bulls with significantly different motility after thawing.     

A Comparative Study of Respiration Rates of Some Aquatic Oligochaetes in Relation to Sublethal Stress

Aquatic oligochaete species selected on the basis of pollution tolerance or intolerance as determined from field studies were used to determine respiration rates under standard conditions, after exposure to sublethal levels of environmental variables (pH, salinity, temperature) or toxicants (Cd, Hg, NaPCP) alone or in pairs. Respiration rates, critical oxygen levels and degree of regulation were not correlated with field-determined tolerance to organic pollution. Stress factors caused increased or lowered respiration rate, loss of regulation (partial or total) and sometimes shifts in critical oxygen level. Regulation was improved in the face of some stresses. Combined stresses produced additional changes in respiration rates and regulation. These responses were complex, which makes them useful indicators of specific stress factors but also indicate complex mechanisms behind the observed changes.     

Diurnal Extension Rates of Wheat Leaves in Relation to Temperatures and Carbohydrate Concentrations of the Extension Zone

The objective of this experiment was to determine how diurnalvariations in rates of leaf extension of wheat plants in anirrigated field crop were related to temperatures and carbohydrateconcentrations of the extension zone. Leaves 3, 4. 5 of themain shoot were studied as each emerged from the encirclingsheath. The carbohydrates in the extension zone of the emergingleaf were analysed by converting them to glucose-6-phosphateand then measuring the reduction of NADP in the presence ofglucose-6-phosphate dehydrogenase. Average hexose concentrations(glucose and fructose) increased each day from 4 up to 5 mgg^–1 fr. wt. and sucrose from 3 up to 7 mg g^–1 fr.wt. with the maximum in mid-afternoon; there were no differencesamong the three leaves. Concentrations of fructans were constantthroughout the day for leaves 3 and 4 but showed a mid-afternoonrise in leaf 5. The average concentrations of fructans in theextension zones increased from 0 to 5 to 11 mg g^–1 fr.wt. for leaves 3, 4, and 5 respectively and was consistent withthe conclusion that there was an increasing over-supply of carbohydratesfor growth of the shoot as the plant increased in size. Ratesof leaf extension were correlated with temperature but not withhexose concentrations. We concluded that the supply of carbohydratesdid not limit the growth of leaves under field conditions buttheir utilization in leaf growth was limited by temperature.The rates of leaf extension increased exponentially with temperatureand the relationship was described by the Arrhenius equation.The Q10 at 15 °C for leaf extension was 2.7 for leaves 4and 5 and 3.2 for leaf 3.