Simultaneous detection of six RNA plant viruses affecting tomato crops using a single digoxigenin-labelled polyprobe

A polyprobe for the simultaneous detection by non-isotopic molecular hybridisation has been developed to detect any of the following six viruses causing important economic losses in tomato crops: Tomato spotted wilt virus, Tomato mosaic virus, Pepino mosaic virus, Cucumber mosaic virus, Potato Y virus and Parietaria mottle virus. The polyprobe detected all six viruses with similar sensitivity to that obtained using individual riboprobes. In addition, we evaluated the possible use of the tissue-printing as a sample preparation technique applied to routine diagnosis of tomato plants with the polyprobe. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effect of Cooling Rate to 5°C,Straw Size and Farm on Fertilizing Ability of Cryopreserved Rabbit Sperm

High fertility and prolificacy in rabbits are currently only achieved using fresh sperm. This study was conducted to determine if the cooling rate to 5°C, the straw size and the farm where artificial inseminations are performed have an impact on the fertilizing ability of rabbit sperm cryopreserved with an extender containing dimethyl sulphoxide (DMSO; 1.75 m ) and sucrose (0.05 m ). Slow cooling to 5°C improved neither fertility rate (58 vs 56% kindling rate for fast and slow cooling, respectively) nor prolificacy (6.5 vs 8.7 total born for slow and fast cooling, respectively; p < 0.05) compared to fast cooling rate to 5°C. The straw size did not have an effect on either fertility or prolificacy (47 vs 57% kindling rate and 6.3 vs 6.8 total born for sperm loaded into 0.25 and 0.5 ml straws, respectively). In addition, similar results were obtained between farms (46–57% kindling rate and 4.9–6.7 total born), although this effect should be studied further. In conclusion, with this extender, slow cooling does not present a beneficial effect on sperm fertilizing ability and either 0.25 or 0.5 ml straws can be used to freeze the sperm, obtaining similar results after artificial insemination. In addition, similar results were obtained between farms when using cryopreserved sperm, and these results were lower than those obtained after artificial insemination with fresh semen. Therefore, new approaches are needed to improve the results obtained when cryopreserved sperms are used before this type of sperm can be used for commercial purposes.     

Characterization of Fusarium mangiferae isolates from mango malformation disease in Southern Spain

During the last years, Fusarium strains have been isolated from shoots and inflorescences of mango trees affected with floral and vegetative malformation in different orchards from the Axarquía region (south of Spain), highlighting the identification of Fusarium mangiferae. With the aim of elucidate epidemiological aspects and design more efficient control strategies, population diversity among the strains of F. mangiferae associated with MMD in Spain was determined by ap-PCR, RAPD-PCR, vegetative compatibility groups (VCGs) and mating type analyses. Three different VCGs were found among the Fusarium mangiferae Spanish isolates, two of them showing similar ap–PCR and RAPD profiles. PCR with primers specific for the mating type (MAT) alleles resulted in amplification of the MAT-2 allele fragment among the majority of the isolates, there being only two isolates MAT-1. This population diversity suggests at least three possible independent introductions of the pathogen into the Axarquía region.     

Classification: Optimum cut-off screen value for foot-and-mouth disease (FMD) immunity evaluation

Circulating antibody levels are used in a dichotomous classification scheme for determining the foot-and-mouth disease (FMD) immune status of cattle. Such a classification scheme may result in 4 categories of animals based on the true state of nature of the animals: the truly protected; the truly unprotected; the falsely protected; the falsely unprotected. The 2 last categories are misclassifications, the frequency of which depend on the specific cut-off value selected for the screening procedure.

A methodology for selecting, from all possible values, the optimal antibody-titer cut-off value is applied in such a way that the number of misclassifications is minimized. This methodology is extended for situations where false protection is considered as being more damaging than false unprotection. Bayes' theorem is used for the evaluation of the performance of the neutralization test, in screening the immune status of cattle. The sensitivity and specificity of the classification scheme, and its ability to predict protection and unprotection are used to evaluate the classification scheme. Methodology is provided for developing estimates of the true FMD protection rate and its variance in cattle populations.     

Computed tomographic retrograde positive contrast cystography and computed tomographic excretory urography characterization of a urinary bladder diverticulum in a dog

A one‐year‐old intact male German shepherd dog was referred with a 3‐month history of dysuria and pollakiuria. Physical examination revealed a large firm mass in the caudal abdomen. Findings from survey radiography, negative contrast cystography, computed tomographic (CT) retrograde positive contrast cystography, and CT excretory urography were consistent with a large urinary bladder diverticulum. An exploratory laparotomy revealed a normal wall appearance in the ventral compartment (true bladder) and marked thinning of the wall in the dorsal compartment (diverticulum). Both ureters inserted into the ventral compartment. The dorsal compartment was excised and histopathology confirmed the diagnosis of urinary bladder diverticulum.     

Serum Levels of Cardiac Markers NT‐proANP and NT‐proBNP in Brachycephalic bitches at Different Gestational Stages

The aim of this study was to determine serum levels of natriuretic peptide precursors (NT‐proANP and NT‐proBNP) during pregnancy in brachycephalic bitches. Fifteen healthy multiparous bitches were selected for this prospective study. Serum levels of NT‐proANP and NT‐proBNP were measured during anoestrous and at 14, 35, 42, 49 and 56 days (2nd, 5th, 6th, 7th and 8th weeks) of pregnancy. Fourteen animals had normal gestations, and one bitch developed single foetus syndrome. The natriuretic peptide levels of this animal were not included in this study; however, it is important to report that its NT‐proANP levels were four times greater than those of normal patients. There was no significant difference (p = 0.072) in NT‐proBNP levels between anoestrous (0.20 ± 0.10 ng/ml) and the different pregnancy weeks (0.27 ± 0.12 ng/ml). There was a positive correlation (p < 0.0001) between NT‐proANP and gestational age, and the levels of this marker increased significantly (p < 0.0001) during the 6th (0.26 ± 0.06 ng/ml), 7th (0.28 ± 0.04 ng/ml) and 8th weeks (0.29 ± 0.05 ng/ml) when compared to anoestrous (0.18 ± 0.02 ng/ml). NT‐proANP serum levels are correlated with gestational development and may be indicative of cardiovascular adaptation in canine brachycephalic pregnancy.     

A single injection of corifollitropin alfa supplemented with human chorionic gonadotropin increases follicular recruitment and transferable embryos in the rabbit

Superovulation protocols are designed to achieve maximum embryo yields. Nevertheless, ovarian response control and the quality of obtained embryos are still a challenge. On the other hand, to save the superovulated embryos until their subsequent use, it is usual to cryopreserve them, so it is also crucial to assess their cryotolerance. The aim of this study was to compare the efficacy of a single injection of corifollitropin alfa (FSH‐CTP) alone or supplemented with human chorionic gonadotropin (hCG) and to determine the impact of this stimulation on in vitro and in vivo development of fresh or devitrified embryos. Our outcomes showed that ovulation rate and recovered embryos were significantly increased when hCG was used. In vitro development of fresh and devitrified embryos and survival at birth were not significantly affected by superstimulation treatment. Results of this study suggest that a single injection of long‐acting FSH‐CTP supplemented with hCG can be effectively used in rabbits to elicit an increase in ovulation rate and number of recovered embryos. Furthermore, we demonstrated that hCG supplementation had no negative effects in embryo cryosurvival and development, showing similar survival rate at birth than FSH‐CTP alone group.     

An assessment of the role played by some oxidation-related aldehydes in wine aroma

The levels of important oxidation-related aldehydes, such as methional, phenylacetaldehyde, (E)-2-hexenal, (E)-2-heptenal, (E)-2-octenal, (E)-2-nonenal, methylpropanal, 2-methylbutanal, and 3-methylbutanal, were determined in 41 different wines belonging to different types (young whites and reds, natural sparkling wines, oxidized young whites and reds, Sherry, aged red wines, Port wines). Except (E)-2-hexenal and (E)-2-heptenal, all of them could be found at levels above threshold. Different compositional patterns were identified: Sherry wines have large amounts of branched aldehydes but not of (E)-2-alkenals, wines exposed to oxygen can have large amounts of (E)-2-alkenals but not of branched aldehydes, while aged wine and Port have relatively large amounts of both classes of compounds. Different sensory tests confirmed the active sensory role of these compounds and revealed the existence of interactions (additive or synergic) between them and with other wine volatiles. (E)-2-Alkenals are related to flavor deterioration, while branched aldehydes enhance dried fruit notes and mask the negative role of (E)-2-alkenals.     

Temporal sequence of cell wall disassembly events in developing fruits. 2. Analysis of blueberry (Vaccinium species)

Softening and pathogen susceptibility are the major factors limiting the marketing of blueberries as fresh fruits, and these traits are associated with fruit cell wall structure. However, few studies that characterize wall modifications occurring during development and ripening have been reported for this fruit. In this study the ripening-associated modifications of blueberry fruit cell walls (composition, pectin and hemicellulose solubilization, and depolymerization) at five stages of ripeness have been analyzed. Xylose was found to be the most abundant noncellulosic neutral sugar associated with fruit walls, and the observed high Xyl/Glc ratio suggested that xylans, which are usually a minor hemicellulosic fruit wall component, are abundant in blueberry. The pectic matrix showed increased solubilization at early and intermediate stages of ripening, but no changes were detected in late ripening. Furthermore, little reduction in pectin polymer size occurred during blueberry ripening. In contrast, hemicellulose levels decreased as ripening progressed, and a clear depolymerization of these components was observed. A model for cell wall degradation in this fruit is discussed.