A NMR study of water distribution in hardwoods at several equilibrium moisture contents

The water state of one tropical (Robinia coccinea) and two temperate (Acer saccharum and Fagus grandifolia) hardwoods was determined at different equilibrium moisture contents (EMC) during desorption at 25°C. NMR technique was used to separate different components of water in wood. The species studied presented different structures, which were apparent on the spin–spin relaxation T₂ values. Three different water components were separated: slow T₂ (liquid water in vessel elements), medium T₂ (liquid water in fiber and parenchyma elements) and fast T₂ (bound or cell wall water). The NMR results showed that even at equilibrated conditions a region exists where loss of liquid water and bound water takes place simultaneously. This region will vary according to the wood structure. Finally, liquid water was present at EMC lower than the fiber saturation point, which contradicts the concept of this point when considered as a bulk property of wood.     

Effects of a plastic cover on soil moisture change in a Mediterranean climatic regime

Water deficit is a serious problem for most agricultural crops, especially in arid–semiarid regions, and limits sustainable development. Production can be improved by reducing evapotranspiration and loss of infiltrated water by the use of a plastic cover. We monitored soil moisture fortnightly over 1 yr using a neutron probe near four trees in an olive grove (Olea europaea, var. Arbequina), two of which had a plastic cover buried in the topsoil around them. These trees were monitored using three neutron probe access tubes per tree to compare the behaviour of soil moisture over time with two other identically instrumented and nearby trees with no cover. Analysis was based on the resultant moisture profiles. The plastic cover retained moisture and increased soil water residence time. During the dry season, the amount of water retained in the plots was at most 15–20% greater in the mid‐season and at least 5–6% greater at the end of season than in the central part of the plots near the trunk. The plastic cover was effective to ca. 50 cm with maximum water content near the soil surface. During the wet season, the cover did not affect soil water. Soil moisture was greater near the tree trunk as a result of stemflow and throughfall.     

Genetic diversity of apricot revealed by a set of SSR markers from linkage group G1

Eight polymorphic simple sequence repeat (SSR) markers located in the G1 linkage group of apricot (Prunus armeniaca L.) were previously developed and evaluated in a small set of cultivars. Those primers were used for studying variability in 77 apricot cultivars belonging to five different geographical groups, such as Chinese, Asian (Irano-Caucasian and Central Asian), North American, Mediterranean and Western European as well as Middle European cultivars. Six of the markers were polymorphic and revealed a total of 71 alleles ranging from 5 (aprigms11) to 20 (aprigms1) alleles per locus with a mean value of 11.83 alleles per locus. In conclusion, the SSR loci located in the G1 linkage group show a level of polymorphism which is similar to loci dispersed throughout the entire genome. The total number of alleles and the number of unique alleles were the highest in Chinese apricots and the lowest in Middle European cultivars. Heterozygosity also showed a decrease from Asia and China to Middle Europe. No association could have been observed between any SSR markers tested and plum pox virus (PPV) resistant phenotype of cultivars. PPV resistant cultivars did not form a separate clade on the dendrogram obtained by UPGMA cluster analysis. Middle European and Chinese cultivars formed separate clusters while other genotypes formed smaller multiple sub-groups or scattered among different clusters. Our results support previous hypotheses on the origin of PPV resistance in North American apricots. The allele data was also presented in a form that allowed the easy observation of allele frequencies in each geographical group at each locus. Using this data field, differences and similarities between cultivar groups can be easily assessed. The analysis demonstrated the links between the North American and Mediterranean apricot germplasm and confirmed that the Chinese and Eastern European cultivars are distantly related.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.     

Supplementation of Citric Acid and Amino Acid Chelated Trace Elements in Low‐Fish Meal Diet for Rainbow Trout Affect Growth and Phosphorus Utilization

This study was conducted to evaluate the effect of citric acid (CA) and amino acid chelated trace elements (AA) on the growth, phosphorus (P) utilization efficiency and loading in rainbow trout fed a diet formulated with a low‐fish meal (FM) concentration. A low‐FM basal diet with or without monocalcium phosphate supplementation served as positive and negative control correspondingly. Dietary treatments consisted of the basal diet supplemented either with 1% CA, AA (equivalent to 40 Zn, 20 Mn, and 4 Cu mg/Kg of diet instead of inorganic trace element mix) or a combination of both. Duplicate groups of 35 fish (13.2 g) were fed until apparent satiation during 12 wk. Group of fish fed diets supplemented with 1% CA or AA had a final growth and feed utilization similar to that observed in fish fed the positive control diet (P > 0.05). P absorption was increased with the addition of CA or AA. P retention efficiency rates for these diets were higher in comparison with the rest of the treatments consequently showing the lowest P loading values (P < 0.05). The results indicate that addition of 1% CA or AA, to a low‐FM diet without inorganic P supplementation enhanced fish growth, P retention, and decreased P load to the environment.     

Species proportions by area in mixtures of Scots pine (<Emphasis Type="Italic">Pinus sylvestris</Emphasis> L.) and European beech (<Emphasis Type="Italic">Fagus sylvatica</Emphasis> L.)

Scots pine (Pinus sylvestris L.) and European beech (Fagus sylvatica L.) dominate many of the European forest stands. Also, mixtures of European beech and Scots pine more or less occur over all European countries, but have been scarcely investigated. The area occupied by each species is of high relevance, especially for growth evaluation and comparison of different species in mixed and monospecific stands. Thus, we studied different methods to describe species proportions and their definition as proportion by area. 25 triplets consisting of mixed and monospecific stands were established across Europe ranging from Lithuania to Spain in northern to southern direction and from Bulgaria to Belgium in eastern to western direction. On stand level, the conclusive method for estimating the species proportion as a fraction of the stand area relates the observed density (tree number or basal area) to its potential. This stand-level estimation makes use of the potential from comparable neighboring monospecific stands or from maximum density lines derived from other data, e.g. forest inventories or permanent observations plots. At tree level, the fraction of the stand area occupied by a species can be derived from the proportions of their crown projection area or of their leaf area. The estimates of the potentials obtained from neighboring monospecific stands, especially in older stands, were poorer than those from the maximum density line depending on the Martonne aridity index. Therefore, the stand-level method in combination with the Martonne aridity index for potential densities can be highly recommended. The species’ proportions estimated with this method are best approximated by the proportions of the species’ leaf areas. In forest practice, the most commonly applied method is an ocular estimation of the proportions by crown projection area. Even though the proportions of pine were calculated here by measuring crown projection areas in the field, we found this method to underestimate the proportion by 25% compared to the stand-level approach.     

Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species

Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n?=?40–46). Three M. hispanica isolates (Hi3/N1, 2n?=?32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n?=?40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200?bp, 955?bp and 399?bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650?bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.     

Fermentation as a bio-process to obtain functional soybean flours

The effect of fermentation on the antioxidant compounds [vitamins C and E, total phenolic compounds (TPC), and reduced glutathione (GSH)], and antioxidant capacity [superoxide anion scavenging activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), inhibition of phosphatidylcholine (PC) peroxidation, and Trolox equivalent antioxidant capacity (TEAC)] of soybean (Glycine max cv. Merit) was studied. Fermentation was carried out in solid state in cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, Bacillus subtilis, and Lactobacillus plantarum and in liquid state either in cracked seeds or milled soybean flours fermented naturally by only the microorganisms present in the seeds or by inoculation with L. plantarum. Vitamin C was not detected in the studied samples. Fermentation caused a decrease in vitamin E activity, except when cracked seed was fermented with A. oryzae, R. oryzae, or B. subtilis that increased 31, 30, and 89%, respectively. Fermentation produced an increase in TPC content and did not affect or reduce the GSH content. Fermentation decreased SOD-like activity drastically, while PRTC increased except when it was carried out naturally in cracked seed. TEAC values rose sharply when soybeans were fermented with B. subtilis. Processed soybean extracts inhibited PC peroxidation in comparison with the control assay. On the basis of the results obtained, the relative contributions of vitamin E, TPC, and GSH to antioxidant capacity were calculated and results showed a very high TPC contribution and a low contribution of GSH and vitamin E activity. Optimum results for functional soybean flours were achieved when fermentation was carried out with B. subtilis inoculum.     

Detection of enzootic nasal tumor virus (ENTV) in a sheep flock in southern Brazil

Enzootic nasal tumor (ENT) is a contagious neoplasm associated with enzootic nasal tumor virus (ENTV), which may induce disease in sheep (ENTV-1) and goats (ENTV-2). This study aimed to describe the occurrence of ENT in two Texel sheep (Ovis aries) from a 75-sheep flock, located in the city of Gravataí, southern Brazil. Animals used to be purchased from different origins, and no specific tests for disease monitoring or quarantine procedure were performed. Affected animals presented respiratory distress, anorexia with severe weight loss, and mucopurulent unilateral nasal discharge. Necropsy was performed in both animals and nasal cavity masses were observed. Histopathological analysis demonstrated an epithelial neoplasm compatible with nasal adenocarcinoma. PCR using a protocol that amplifies a 591 bp sequence of 5’LTR-gag region of ENTV1 was performed followed by DNA sequencing. Both samples were positive, and the sequences obtained presented highest identity (97%) with ENTV strain TN28 (GenBank accession number MH899613) detected in a Texel sheep from Scotland. This is the first report of ENTV-1 leading to enzootic nasal tumor in sheep in Latin America, which confirms the presence of the retrovirus in sheep flocks in the Brazilian territory.