Comparison of image analysis and direct measurement of UPOV taxonomic characteristics for variety discrimination as determined over five growing seasons, using industrial chicory as a model crop

Tests for distinctness, uniformity and stability (DUS) are an essential component of variety registration and granting plant breeders’ rights. This study, performed on industrial chicory (Cichorium intybus L.), included (1) an evaluation of the discriminating power of the UPOV characteristics and (2) a comparison of the root shape between the UPOV-based characteristics and characteristics determined via image analysis. Five consecutive growing periods were studied to assess the long-term stability of the parameters. Lack of discriminating power of characteristics for examining distinctness may impede breeding progress by failing to discriminate novel candidate varieties. However, a balance needs to be set so that newly introduced characteristics or new evaluation methodologies are not too powerful as to damage the protection of registered varieties. The results showed great differences in the discriminating power of the UPOV characteristics. The elimination of some of these low-discriminating characteristics would probably result in savings, but would have no consequences on the decisions taken by DUS testing authorities. Image analysis-derived characteristics could either replace currently-used parameters or provide additional characteristics with good discriminating power, determined in an objective and standardized way. Here we discuss possible implications for variety registration.     

Small Demodex populations colonize most parts of the skin of healthy dogs

Background – It is unproven that all dogs harbour Demodex mites in their skin. In fact, several microscopic studies have failed to demonstrate mites in healthy dogs. Hypothesis/Objectives – Demodex canis is a normal inhabitant of the skin of most, if not all, dogs. This hypothesis was tested using a sensitive real‐time PCR to detect Demodex DNA in the skin of dogs. Animals – One hundred dogs living in a humane society shelter, 20 privately owned and healthy dogs and eight dogs receiving immunosuppressive or antineoplastic therapy. Methods – Hair samples (250–300 hairs with their hair bulbs) were taken from five or 20 skin locations. A real‐time PCR that amplifies a 166 bp sequence of the D. canis chitin synthase gene was used. Results – The percentage of positive dogs increased with the number of sampling points. When a large canine population was sampled at five cutaneous locations, 18% of dogs were positive for Demodex DNA. When 20 skin locations were sampled, all dogs tested positive for mite DNA. Our study indicates that Demodex colonization of the skin is present in all dogs, independent of age, sex, breed or coat. Nevertheless, the population of mites in a healthy dog appears to be small. Demodex DNA was amplified from all 20 cutaneous points investigated, without statistically significant differences. Conclusions and clinical importance – Using a real‐time PCR technique, Demodex mites, albeit in very low numbers, were found to be normal inhabitants of haired areas of the skin of healthy dogs.     

Isolation and characterisation of Shiga toxin-producing Escherichia coli O157:H7 from calves in Argentina

Four hundred and twenty-two calves were examined for intestinal carriage of Shiga toxin-producing Escherichia coli O157:H7 using conventional plating. Two (0.5%) E. coli O157 were recovered. They were compared with 96 Argentine strains of different origin by pulsed-field gel electrophoresis, phage typing and PCR-RFLP of stx2 genes. One strain isolated from a calf, was closely related with 18 strains of clinical origin.     

Effects of mycorrhizal inoculation of shrubs from Mediterranean ecosystems and composted residue application on transplant performance and mycorrhizal developments in a desertified soil

Arbuscular mycorrhizal inoculation and composted residue application are being assayed to aid restoration of desertified areas under Mediterranean climate. The particular objective of the present study was to assess the short-term (8 months) effects on the initial stages of plant performance and on mycorrhizal propagule release, key factors to decide further developments in the restoration process. Mycorrhizal inoculation, with Glomus intraradices, was practised during nursery production of representative shrub species from Mediterranean ecosystems and composted residues were added to soil before transplanting to a desertified area in southern Spain. Pistacia lentiscus, Rhamnus lycioides, Olea europaea subsp. sylvestris and Retama sphaerocarpa, key species from the natural succession in the target area, were the test plants. Mycorrhizal inoculation, and in some cases compost addition, improved the ability for nutrient acquisition by plants upon transplanting in the field. The number of "infective" mycorrhizal propagules was higher in soil around mycorrhiza-inoculated shrubs than that around the corresponding non-inoculated controls. The organic amendment significantly increased propagule production in the rhizosphere of mycorrhiza-inoculated plants. The number of mycorrhizal spores was relatively low in soil around transplants, being hardly affected by treatments. Only three distinguishable glomalean spore morphotypes were found, belonging to the species Glomus geosporum, G. contrictum and Scutellospora calospora, with very few unidentified spores, corroborating the low diversity in degraded ecosystems. An increased development of the extramatrical AM mycelium was found in soil around the roots of the four mycorrhiza-inoculated test plants, probably the main source of AM fungal propagules in the ecosystem at this stage of plant development. In conclusion, the tailored AM inoculation assayed was functioning under field conditions to enhance nutrient acquisition by the target indigenous shrubs and, in interaction with organic amendments, promoted mycorrhizal propagule production in soil, critical factors to benefit further stages of the revegetation process.     

Resveratrol content of Palomino fino grapes: influence of vintage and fungal infection

The objective of this study was to determine the influence of certain factors on the resveratrol content of Palomino fino grapes, cultivated in the Jerez-Xérèz-Sherry area, at the moment of harvest. The results show that the resveratrol content is highly influenced by the climatic conditions prior to the period of maturation of the fruit. On the other hand, the gray mold pressure in the vineyard, a fungal infection caused by Botrytis cinerea, increased the resveratrol contents at the early stages of fungal development. When Botrytis development was extensive, the resveratrol content tended to decrease in the juice but tended to increase in the skin. Physiological stress of the plant leads to increases in the resveratrol content, caused as much by the climatic conditions of the vintage as by biotic factors. In this case resveratrol is present mainly in the glycosylated form.     

Improving seed yield in red clover through marker assisted parentage analysis

Red clover (Trifolium pratense L.) is an important perennial forage crop in Europe. Unfortunately, seed production of modern cultivars is often unsatisfactory and breeding progress for higher seed yield is slow. We evaluated whether marker assisted parentage analysis could identify progeny plants with two high seed-yielding parents in a diploid selection trial. Furthermore, we studied if breeding for high seed yield could be speeded up by selecting progeny plants with two high seed-yielding parents, compared to traditional selection on the mother plant only. In this study, 111 genotypes with excellent vegetative traits from a first cycle selection trial were allowed to pollinate each other in isolation. After seed harvest, ten high seed-yielding plants were identified and their seeds were used to grow ten half-sib progeny populations, with 1,121 plants in total. Two multiplex primer sets targeting 18 SSR loci were designed and used to fingerprint parental and progeny plants. Parentage was reliably determined for 1,083 progeny plants: 135 plants were the result of a cross between two high seed-yielding parents. In a 2-year seed yield trial, the seed yield of these 135 progeny plants was compared to that of a control group of 216 progeny plants from the same ten high seed-yielding mother plants but with no selection on the male contribution, as is current practice. Compared to the control group, progeny plants with two high seed-yielding parents had significantly higher seed yields, 23.0 and 75.9 % higher in 2012 and 2013 respectively, and a significantly higher thousand seed weight, 6.6 and 5.7 % higher in 2012 and 2013 respectively. Seed yield was strongly correlated with flower abundance (r = 0.643) and plant volume (r = 0.593 in 2012 and r = 0.509 in 2013) and negatively correlated with susceptibility to mildew (r = ?0.145). We conclude that breeding programs aiming at increasing seed yield in diploid red clover cultivars would strongly benefit from the integration of marker assisted parentage analysis to identify those progeny plants that are the result of a cross between two high seed-yielding plants. This goal cannot be achieved with traditional family selection where only the seed yield of the mother plant is known.     

A long-term in vitro culture of bovine epithelial cells on collagen rafts

In the present work, we established and characterized a 3D functional polarized primary bovine oviduct epithelial cells (BOECs) culture on free-floating type I collagen hydrogels (rafts) at an air-liquid interface (ALI). Intercellular junctions, ultrastructural cellular morphology and the expression of the OVGP1 closely recapitulated those of the in vivo epithelium lining. These morphological and physiological epithelial cell features were maintained under standard DMEM/F12 with 10% foetal bovine serum culture medium for at least 28 days of ALI culture. The versatility of the BOECs raft cultures should allow testing of toxicity compounds, in vitro evaluation of physiological or pathological oviductal states, and the study of epithelial-mesenchymal interactions that are critical for the maintenance of oviductal homeostasis.     

Subclinical ketosis in dairy cows: prevalence and risk factors in grazing production system

The purpose of this study was to estimate the prevalence of Subclinical ketosis (SCK) between 4 and 19 days in milk (DIM) in a grazing production system and investigate the importance of potential risk factors for SCK. This cross‐sectional study was conducted in dairy cows (n = 107), which had more of two parities. The concentration of β‐hydroxybutyric (BHB) in blood was quantified through a hand‐held meter. Potential risk factors evaluated were calving interval (CI), milk yield in previous lactation, metritis, dystocia, calf sex (male), parity (≤3 vs. ≥4) and pre‐partum body condition score (BCS ≤ 3.5 vs. ≥3.75). Prevalence of SCK was 10.3% (95% CI 4.7–15) between 4 and 19 DIM. Risk factors identified were the occurrence of both metritis and pre‐partum BCS ≥ 3.75. Cows with metritis had 4.9 (95% CI 1.17–20.98) times more risk of developing SCK than cows without metritis. And the cows with pre‐partum BCS ≥ 3.75 had 5.25 (95% CI 1.32–21.11) times more risk of developing SCK than cows with pre‐partum BCS ≤ 3.5. Metritis could induce a lower feed intake and promote the development of SCK. High pre‐partum BCS could induce a greater mobilization of body reserves altering liver function and aggravating post‐partum NEB. The results are indicative of the expected prevalence of SCK in grazing production system. Factors associated could help to identify cattle at risk of SCK and improve the management of strategies to limit the effects.     

Quantitative determinations of chemical compounds with nutritional value from inca crops:Chenopodium quinoa (‘quinoa’)

Quantitative determinations of total and soluble proteins, total and free sugars, starch, total lipids, tanins, ash (Ca, Na, K, Fe, and P), and caloric value were carried out on quinoa flour.Results show that the amount of soluble proteins was higher than the standard value for wheat and maize and was very close to that of barley's. The yield of free sugars like glucose (4.55%), fructose (2.41%) and sucrose (2.39%) were also of importance.Iron and calcium levels were higher than the reported values for maize and barley. The same occurred for the caloric value (435.5 Kcal/100 g).The content of saponins was also examined since its effect on red blood cells of group A and O has been related as a potential problem of the Andes population.From the chemical analysis a more complete view about quinoa as human food was presented.     

Antagonistic effect of gibberellic acid and boron on protein and carbohydrate metabolism of soybean germinating seeds

Lots of soybean seeds (Glycine max L. Merr. cv MG 13 G2) were pretreated with solutions of boron (1 or 2 μg/ml) or/and gibberellic acid (GA₃) (0.1 μg/ml) during three hours, and germinated in a culture chamber at 24°±1°C. Germination and seedling growth were inhibited by boron. GA₃ activated both processes and partially reversed the inhibitory effect of the microelement. This protective effect of GA₃ against the negative action of boron is particularly due to the antagonism GA₃ /boron on protein and carbohydrate metabolism during the seed germination process.