Skeletal muscle fiber size in untrained and endurance-trained horses.

The mean area and minimal diameter of 3 histochemically determined myofiber types (1, 2A, and 2B; myosin ATPase in acid buffer) were calculated in middle gluteal muscle biopsy specimens from 62 stallions, 47 Andalusians and 15 Arabians, ranging in age from 6 to 12 years. Fourteen Andalusians and 7 Arabians were untrained, and the remainder were actively endurance-trained. The 6-month training schedules involved walking, slow trotting, and cantering. Fourteen Andalusians were moderately endurance-trained, whereas the other 19 Andalusians and 8 Arabians were strongly endurance-trained. Significant differences were not recorded between untrained and endurance-trained Arabians with respect to the area (type 1, 3,194 +/- 869 microns 2 and 3,150 +/- 370 microns 2; type 2A, 3,819 +/- 890 microns 2 and 3,380 +/- 356 microns 2; and type 2B, 4,872 +/- 962 microns 2 and 4,417 +/- 646 microns 2) or minimal diameter (type 1, 52.2 +/- 7.4 microns and 52.8 +/- 3.1 microns; type 2A, 58.1 +/- 6.7 microns and 55.0 +/- 2.8 microns; and type 2B, 65.3 +/- 6.4 microns and 63.4 +/- 4.3 microns) of the 3 fiber types, nor between untrained and endurance-trained Andalusians with respect to the area (untrained, 3,990 +/- 690 microns 2; moderately endurance-trained, 3,882 +/- 347 microns 2; and strongly endurance-trained, 3,758 +/- 510 microns 2) and minimal diameter (untrained, 58.1 +/- 4.7 microns; moderately endurance-trained, 59.7 +/- 2.7 microns; and strongly endurance-trained, 58.7 +/- 4.5 microns) of 2A fibers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Determination of sulfamethazine and trimethoprim in liquid feed premixes by HPLC and diode array detection, with an analysis of the uncertainty of the analytical results.

Sulfamethazine (SMZ) and trimethoprim (TMP) are antibacterials used in veterinary practice. This paper describes a method for their determination in veterinary liquid feed premixes that is based on liquid chromatography with diode array detection. Gradient elution with methanol and ammonium acetate achieved excellent separation of the two analytes within 15 min without any interference from the matrix. Absorbance of the column effluent was monitored at 264 nm for SMZ and at 230 nm for TMP. Detailed analyses of the uncertainties of determinations afford estimated expanded uncertainties of, respectively, 0.2 and 0.1 w/v % for typical SMZ and TMP concentrations of 10.7 and 2.1 w/v %, respectively. At the lower end of the calibrated range of the method, the dominant source of uncertainty is the preparation of standards and the construction of the calibration line.     

Pregnancy Rates in Repeat-breeder Heifers Following Multiple Artificial Inseminations during Spontaneous Oestrus

Hormonal asynchronies during oestrus, related to the presence of suprabasal plasma-progesterone (P4) concentrations and a delayed ovulation, interfere with the fertility of repeat-breeder heifers (RBH). Since tubal dysfunction can occur in connection with hormonal asynchronies and constrained availability of fertile spermatozoa at the time of ovulation, the present study tested the hypothesis that frequent sperm deposition from onset of oestrus to ovulation may improve pregnancy rates in RBH. Five RBH and five virgin heifers (VH; controls) were repeatedly artificially inseminated (AI) at 6 h intervals from onset of oestrus to spontaneous ovulation. Hormone analyses revealed suprabasal P₄ concentrations and a delay in the occurrence of the luteinising hormone (LH) surge, but a normal cortisol profile in RBH. Compared with controls, RBH presented longer interval from onset of oestrus to ovulation, and therefore, received more AIs. Pregnancy rates in RBH reached control levels (60%; NS), indicating that the hypothesis might be correct. Pregnancy rates in VH were below the expected range, presumably attributed to a deleterious influence of the frequent handling. The study suggests that pregnancy rates can be improved in RBH by frequent AI in relation to spontaneous ovulation. However, this practice of repeated manipulations, while seeming not to show adverse effects, lacks practicality for routine use.     

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Physiological Response of Multiple Contrasting Rice (Oryza sativa L.) Cultivars to Suboptimal Temperatures

The physiological response of multiple rice cultivars, eighteen initially and eight cultivars later on, to suboptimal temperatures (ST) conditions was investigated in laboratory and outdoor experimental conditions. Treatment with ST decreased growth in different extents according to the cultivar and affected the PSII performance, determined by chlorophyll fluorescence fast‐transient test, and stomatal conductance, regardless the experimental condition. Two groups of cultivars could be distinguished on the base of their growth and physiological parameters. The group of cultivars presenting higher growths displayed optimal JIP values, and higher instantaneous water use efficiency (WUE_i), due to a lower Gs under ST, unlike cultivars showing lower growth values, which presented worse JIP values and could not adjust their Gs and hence their WUE_i. In this work, we detected at least two cultivars with superior tolerance to ST than the cold tolerant referent Koshihikari. These cultivars could be used as parents or tolerance donors in breeding for new crop varieties. On other hand, positive and significant correlations between data obtained from laboratory and outdoor experiments suggest that laboratory measurements of most of the above mentioned parameters would be useful to predict the response of rice cultivars to ST outdoor.     

Seed Shattering of Cañahua (Chenopodium pallidicaule Aellen)

Cañahua (Chenopodium pallidicaule Aellen) is a semi‐domesticated relative of quinoa (Chenopodium quinoa Willd.) with high nutritious quality. It is tolerant to frost, drought, saline soils and pests. One seed yield limitation is seed loss during the maturity stages. Two greenhouse experiments in Denmark and field experiments in Bolivia were carried out to determine seed shattering in landraces and cultivars with different growth habits. 15–21 % of the seed shattering in the fields took place whilst the plants still were flowering and 25–35 % during physiological maturity. Seed shattering varied between locations on the Bolivian Altiplano. Cañahua types with the semi‐prostrate growth (‘lasta’) had the highest seed shattering rate in the greenhouse experiments. The Umacutama landrace had lower seed shattering (1 %) than the cultivar Kullaca (7.2 %) both of the ‘lasta’ type. Under field conditions, the cultivar Illimani with the erect growth (‘saihua’) had the highest seed shattering rate (6.4–33.7 %) at both locations and at four different sowing dates. The Umacutama had the lowest rate (0.5–1.5 %). There were no significant differences between plants of the ‘lasta’ and the ‘saihua’ types. The landrace had significantly less seed loss than the cultivars. However, in the greenhouse, the landrace yield was approximately 25 % lower than the yields of the cultivars. In general, cañahua cultivars had higher yield compared to landraces, but also a higher seed shattering rate. Landraces may be used in breeding programmes to develop high‐yielding cultivars with reduced seed shattering.     

Application to soil of disinfectants through irrigation reduces Verticillium dahliae in the soil and verticillium wilt of olive

The application of disinfectants through drip irrigation could be a feasible practice against verticillium wilt (Verticillium dahliae) of olive. OX-VIRIN (activated peroxide) and OX-AGUA AL25 (quaternary ammonium compounds) are two disinfectants that have shown efficacy against V. dahliae in irrigation water and potential for reducing the disease in young olive plants. In this work, various post-planting application strategies incorporating OX-VIRIN (once a month, or twice a month on alternate or successive weeks) or OX-AGUA AL25 (once a month, or twice a month on alternate weeks) were assessed for their effect on V. dahliae in soil, disease in olive trees, and olive yield, in a 2-year pot-experiment under natural environmental conditions. The disinfectants were injected via metering pumps into a drip irrigation system that irrigated olive trees planted in V. dahliae-inoculated soil. All the application strategies significantly reduced the total inoculum density in soil compared to controls with no disinfectants and noninoculated soil. The microsclerotia density was also significantly reduced in disinfested soils by 73.6–86.8%, depending on the strategy. The symptoms and infection incidence were always lower in treatments subjected to disinfestation. The treatment with OX-AGUA AL25 applied twice a month on alternate weeks most reduced the symptoms (by 53.0%) and colonization index (by 70.8%) with respect to untreated water control. This soil disinfestation also significantly strengthened the symptom remission. Tree growth and production were negatively affected by soil inoculation (reduced by 45.6% and 88.7%, respectively), but not so by disinfectants, which even relieved the reduction in inoculated soils, especially when OX-AGUA AL25 was applied.     

Salmonid fish viruses and cell interactions at early steps of the infective cycle

A flow cytometric virus-binding assay that directly visualizes the binding and entry of infectious pancreatic necrosis virus (IPNV), infectious haematopoietic necrosis virus (IHNV) and virus haemorrhagic septicaemia virus (VHSV) to several cell lines was established. The highest efficiency of binding was shown by the BF-2 cell line and this was used to study, at the attachment level, the interactions of these cells with salmonid fish viruses in coinfections, and to further determine if the earliest stage of the viral growth cycle could explain the previously described loss of infectivity of IHNV when IPNV is present. Our results demonstrated that IPNV binds to around 88% of cells either in single or dual infections, whereas IHNV attachment always decreased in the presence of any of the other viruses. VHSV binding was not affected by IPNV, but coinfection with IHNV reduced the percentage of virus-binding cells, which suggests competition for viral receptors or co-receptors. Internalization of the adsorbed IHNV was not decreased by coinfection with IPNV, so the hypothetical competence could be restricted to the binding step. Treatment of the cells with antiviral agents, such as amantadine or chloroquine, did not affect the binding of IPNV and VHSV, but reduced IHNV binding by more than 30%. Tributylamine affected viral binding of the three viruses to different degrees and inhibited IPNV or IHNV entry in a large percentage of cells treated for 30 min. Tributylamine also inhibited IHNV cytopathic effects in a dose-dependent manner, decreasing the virus yield by 4 log of the 50% endpoint titre, at 10 mm concentration. IPNV was also inhibited, but at a lower level. The results of this study support the hypothesis that IHNV, in contrast to VHSV or IPNV, is less efficient at completing its growth cycle in cells with a simultaneous infection with IPNV. It can be affected at several stages of viral infection and is more sensitive to the action of antiviral compounds.