Dimethylformamide Improves the In vitro Characteristics of Thawed Stallion Spermatozoa Reducing Sublethal Damage

A total of 42 ejaculates were used in the experiment; six ejaculates per stallion, obtained from seven Pure Spanish stallions (PRE), were split and frozen in freezing media with different concentrations and combinations of cryoprotectant (CPA): (i) Cáceres (skim milk based extender) containing 2.5% glycerol (2.5GL), (ii) Cáceres containing 1.5% glycerol and 1.5% dimethylformamide (1.5%GL–1.5%DMFA), (iii) Cáceres extender supplemented with 1.5% glycerol and 2.5% dimethylformamide (1.5%GL–2.5%DMFA) and (iv) Cáceres extender supplemented with 4% dimethylformamide (4%DMFA). After at least 4 weeks of storage in liquid nitrogen (LN), straws were thawed and semen analysed by computer‐assisted sperm analysis and flow cytometry (membrane lipid architecture (Merocyanine 540), integrity and sublethal damage (YoPro‐1) and mitochondrial membrane potential (JC‐1)). After thawing, better results were observed in samples frozen in 4%DMFA or in combinations of 1.5%GL–2.5%DMFA, in fact total motility increased by 16% in the 4%DMFA group compared to 2.5%GL (P < 0.05). Also, there was an increment in the percentage of progressive motile sperm in the 1.5%GL–2.5%DMFA group (9.8% 2.5GL vs 19% in the 1.5%GL–2.5%DMFA group p < 0.05); also, samples frozen in the 4%DMFA group had more intact (YoPro‐1 negative) sperm post‐thawing, 29.3% in 2.5%GL vs 36.7% in 4%DMFA group (p < 0.05). Membrane lipid architecture was not affected by any of the cryoprotectants tested, while samples frozen in 4%DFMA had a lower percentage of mitochondria with lower membrane potential. It is concluded that DMFA improves the outcome of cryopreservation of stallion spermatozoa mainly reducing sublethal cryodamage.     

Genotyping Shorthorn cattle for generalised glycogenosis

OBJECTIVE: To develop a procedure for routine genotyping of Shorthorn cattle for the generalised glycogenosis allele in exon 18 of the acidic alpha-glucosidase gene. PROCEDURE: Allele-specific amplification and double mismatch amplification procedures for the discrimination of the exon 18 alleles were evaluated using leucocytes and hair roots as sources of target DNA. RESULTS: Allele-specific amplification was effective for genotyping Shorthorn cattle at the 2454 site when purified DNA was used as target for the polymerase chain reaction. However, when the target DNA was derived from hair roots, differences in the relative yield of wild-type and mutant amplicons were observed. The double mismatch amplification procedure was effective in genotyping all subjects, independent of the source of DNA. The unique cleavage sites for Drd I and PshA I within exon 18 are present and absent respectively in the wildtype amplicon, and are lost and acquired, respectively, in the mutant amplicon. In addition, the Drd I and PshA I mismatching cleavage sites incorporated into the primers serve as internal controls for Drd I and PshA I cleavage. CONCLUSION: The double Drd I/PshA I mismatch amplification procedure using hair root samples as the source of DNA is a robust method for genotyping Shorthorns for generalised glycogenosis.     

Inoculum Densities of Fusarium oxysporum f. sp. vasinfectum and Meloidogyne incognita in Relation to the Development of Fusarium Wilt and the Phenology of Cotton Plants (Gossypium hirsutum)

ABSTRACT Development of Fusarium wilt in upland cotton (Gossypium hirsutum) usually requires infections of plants by both Meloidogyne incognita and Fusarium oxysporum f. sp. vasinfectum. In this study, the soil densities of M. incognita and F. oxysporum f. sp. vasinfectum and the incidence of Fusarium wilt in three field sites were determined in 1982-1984. Multiple regression analysis of percent incidence of Fusarium wilt symptoms on population densities of M. incognita and F. oxysporum f. sp. vasinfectum yielded a significant fit (R (2) = 0.64) only on F. oxysporum f. sp. vasinfectum. Significant t-values for slope were also obtained for the interaction of M. incognita and F. oxysporum f. sp. vasinfectum, but densities of M. incognita and F. oxysporum f. sp. vasinfectum were also related on a log(10) scale. The physiological time of appearance of first foliar symptoms of Fusarium wilt, based on a degree-days threshold of 11.9 degrees C (53.5 degrees F), was used as a basis for determining disease progress curves and the phenology of cotton plant growth and development. Effects of Fusarium wilt on plant height and boll set were determined in three successive years. Increases in both of these plant characteristics decreased or stopped before foliar symptoms were apparent. Seed cotton yields of plant cohorts that developed foliar wilt symptoms early in the season (before 2,000 F degree-days) were variable but not much different in these years. This contrasted with cohorts of plants that first showed foliar symptoms late in the season (after 2,400 F degree-days) and cohorts of plants that showed no foliar symptoms of wilt. Regression analyses for 1982-1984 indicated moderate to weak correlations (r = 0.16-0.74) of the time of appearance of the first foliar symptoms and seed cotton yields.     

Resident forum abstractsEVALUATION OF THROMBOELASTOGRAPHY (TEG) IN NORMAL CATS

Objective: To establish normal parameters of thromboelastography (TEG) in healthy adult cats. Background: Thromboelastography (TEG) is an in vitro test of coagulation that has been shown to be useful in humans, dogs and select species to identify and quantify alterations of hemostasis (e.g., hypercoagulable and hypocoagulable states). It has also been demonstrated to be useful in monitoring effects of anticoagulant therapies. This test has not been evaluated in cats. Methods: Blood was collected from 25 clinically normal cats by venipuncture using a 21 gauge×3 1/2 inch butterfly catheter and syringe for medial saphenous or jugular venipuncture. A single 1.8 mL sample in 3.8% Sodium Citrate (9:1) was collected from each cat. Recalcified whole blood was analyzed 30 minutes following collection with the TEG® 5000 analyzer (Haemoscope, Niles, IL). Analysis temperature was 37.6°C. TEG parameters recorded included: R‐value (represents initial fibrin formation), K (time from R to standard fixed measure of clot firmness which represents contributions of platelets and fibrinogen), maximum amplitude (MA; represents absolute clot strength), and alpha angle (α; the slope of TEG tracing which represents rate of clot formation). The coagulation index (CI) was derived from the formula generated for humans to provide an overall assessment of whether the sample was hyper‐ or hypocoagulable. Results: Values for the 25 normal cat samples are reported as mean ±2 standard deviations. R=2.97; 1.23–4.72; K=1.54, 0.38–2.71; α=70.70, 57.76–83.65; MA=58.50, 45.26–71.74 and CI=2.27, 0.07–4.46. Compared to historical information obtained on normal dogs, cats have significantly shorter R and K and larger α, MA and CI. Conclusions: TEG does have reproducible performance when used to evaluate coagulation status in normal cats. Compared to dogs, normal cats favor a hypercoagulable state. Species‐specific normal values are necessary for interpretation of TEG results. This test bears potential value for use in future experimental and clinical work to investigate hemostasis in cats receiving anticoagulant therapies or in cats suffering from diseases such as cardiomyopathy which are thought to be associated with altered coagulation status.     

甜菜夜蛾综合防治技术研究

甜菜夜蛾是一种多食性、暴食性害虫,其暴发成灾将给农业生产造成巨大损失.防治甜菜夜蛾,应加强预测预报,采取农业、物理、生物、化学和成虫诱杀等防治手段相结合的综合技术,在发生关键时期用药防治,可取得较好的防治效果.     

花椒吹绵蚧农药防治适期研究

用45%晶体石硫合剂、40%杀扑磷乳油、24%虱蚧净乳油、2.5%敌杀死乳油、40%氧化乐果乳油5种农药,在花椒吹绵蚧休眠期和若虫活动游走期进行农药防治试验。结果表明,在花椒吹绵蚧休眠期用45%晶体石硫合剂防治的最佳时期为3月中旬;花椒吹绵蚧若虫活动游走期,农药防治最佳时期为4月中旬,防治效果依次为40%杀扑磷乳油>24%虱蚧净乳油>2.5%敌杀死乳油>40%氧化乐果乳油。     

Effects of stocking on the genetic structure of brown trout,Salmo trutta,in Central Europe inferred from mitochondrial and nuclear DNA markers

Abstract Stocking has had a considerable effect on wild brown trout, Salmo trutta L., populations throughout Europe. To elucidate this impact and to outline further management strategies, the genetic structure of 25 wild populations and five hatchery stocks from Czech Republic and Slovakia were analysed using mitochondrial (control region) and nuclear DNA (microsatellites, LDH‐C1*) markers. Stocking practices have caused massive hybridisation between the Atlantic and Danube brown trout strains in the central Danube basin and have lead to a loss of among‐population divergence in Slovakia and the eastern part of Czech Republic. Comparison with studies from neighbouring countries revealed substantial differences in haplotype, allele frequencies and genetic diversity across Central Europe. Differences in stocking management and origin of breeding stocks appear to be crucial factors for the spatial variability of the genetic structure of brown trout.     

<Emphasis Type="Italic">Pulque</Emphasis>, An Alcoholic Drink from Rural Mexico,Contains Phytase. Its <Emphasis Type="Italic">in vitro</Emphasis> Effects on Corn Tortilla

Pulque is made by fermenting the agave sap or aguamiel of Agave atrovirens with a whole array of microorganisms present in the environment including several lactic acid bacteria and yeasts such as Saccharomyces cerevisiae. Ascorbic acid was determined in pulque and aguamiel, respectively. Phytase activity in lees, liquid and freeze-dried pulque was assayed by measuring the appearance of phosphate from phytate by a colorimetric method likewise phosphate from phytate present in fresh corn tortilla was measured after in vitro incubation with pulque. Iron, zinc, calcium, magnesium and selenium contents were measured in pulque and corn tortilla as well as in nixtamalized corn flour (NCF), the latter is used to make instant tortilla, since corn provides most of the energy as well as most of the phytate in the Mexican rural diet. Pulque showed phytase activity but much less ascorbic acid and iron than previously reported; additionally, phytase in pulque hydrolyzed most of phytate’s corn tortilla. Lees, which is mostly made of pulque’s microbiota, significantly accumulated iron and zinc but no selenium. NCF was fortified with iron by the manufacturers but poorly blended. There were significant differences on selenium content between tortillas samples, apparently some soils in central Mexico are selenium deficient. Moderate pulque intake appears to increase the bioavailability of iron and zinc bound by phytate in corn.     

Effect of steel-making slag as a soil amendment on arsenic uptake by radish (Raphanus sativa L.) in an upland soil

The steel-making slag (SMS), a by-product of steel manufacturing process with an alkaline pH (11–12) and high amount of iron (Fe) and calcium (Ca) oxides, was used to reduce arsenic (As) phytoextractability. The by-product was selected as an alternative to commercial Fe oxides, which can decrease plant uptake, but they are expensive if used as amendments of contaminated arable soils. SMS was applied at rates 0, 2, 4, and 8 Mg ha⁻¹ to an As (1 N HCl-extractable As 25 mg kg⁻¹) contaminated soil prepared by mixing non-contaminated soil and mine tailings and cropped to radish (Raphanus sativa L.) seeding. Calcium hydroxide (Ca(OH)₂), a common liming material in Korea, was applied at the same rates for comparison. Steel-making slag more effectively suppressed radish As uptake and increased yield than Ca(OH)₂ due to stronger As immobilization because it significantly increased extractable Fe concentration and decreased extractable As. The SMS-treated soil showed an apparent increase in As chemisorbed by Fe and Al oxides and hydroxides of surface soil, As associated at the Fe and Al oxides and hydroxides of internal surfaces of soil aggregates, and Ca-associated As. The steel-making slag can be a good soil amendment to suppress As phytoextractability and improve nutrient balance in As-contaminated soil.     

Marker-assisted selection in faba bean (Vicia faba L.)

Among the multiple applications of DNA markers in breeding, the most promising for cultivar development is marker-assisted selection (MAS). Genetic improvement by MAS has been carried out with success in several legume crops such as soybean, common bean and pea, however, in other species such as faba bean it is still in its early stages. This article provides an overview of the genomic resources and molecular markers currently available in faba bean, with an emphasis on development and application of MAS for genetic improvement of the crop. In general, genetically simple traits have received more attention than genetically complex characters encoded by multiple genes. Research has mainly focussed on developing molecular markers for selecting resistance to a parasitic weed and other major diseases. As a result, molecular breeding for resistance to crenate broomrape, ascochyta blight, rust and chocolate spot is underway, and promising results have been obtained. Recently, markers linked to a gene controlling growth habit or to select against traits affecting the nutritional value of seeds (tannins, vicine and convicine content) have also been reported, which may facilitate a more efficient selection of new cultivars free of anti-nutritional compounds. In the near future, molecular markers should be developed for many other highly sought-after traits that are difficult to breed conventionally such as frost or drought tolerance. Comparative genomics and synteny analyses with closely related legumes, together with extensive mapping of resistance gene analogs (RGAs), will reveal new candidate genes and selectable markers for use in MAS. Finally genomic tools such as macro- and microarrays may eventually become available for use in faba crop improvement.