Chestnut and mimosa tannin silages: Effects in sheep differ for apparent digestibility, nitrogen utilisation and losses

This paper reports effects of chestnut and mimosa tannins on N utilisation in sheep. Tannins were added to grass either at ensilage or incorporated into grass silage at feeding. The study used an 8 × 5 incomplete Latin Square design with eight mature wether sheep and five 21-day periods. Tannin additions reduced in vivo apparent digestibilities of dry matter (DM), organic matter (OM) and neutral detergent fibre (aNDFom) compared with the untreated control silage (P<0.001). Reductions ranged from 7.6% for DM to 8.5% for aNDFom. Chestnut compared to mimosa tannin silages produced higher values for DM intake (734 g/day versus 625 g/day) and in vivo digestibility for DM, OM and aNDFom (0.66, 0.68 and 0.69 versus 0.61, 0.63 and 0.62; P<0.001). A substantial shift occurred in the pattern of N excretion in sheep fed the tannin versus control silages. As a proportion of daily N intake, urinary N losses were lower (56.4 g/100 g N versus 65.1 g/100 g N intake) and faecal N losses were higher (40.2 g/100 g N versus 29.8 g/100 g N intake) for sheep fed the tannin silages compared with those fed the control grass silage (P<0.001). Nitrogen intake was higher in sheep fed the chestnut compared to mimosa tannin silages (16.2 g/day versus 13.4 g/day; P<0.001), reflecting the lower DM intake of sheep fed the mimosa silages. However, faecal N loss was lower for chestnut compared to mimosa tannin silage fed sheep (38.2 g/100 g N versus 42.1 g/100 g N intake; P<0.01), resulting in higher N retentions with the chestnut compared to the mimosa silage fed sheep (5.49 g/100 g N versus 1.38 g/100 g N intake). Faecal N losses were also higher when tannins were added during ensiling rather than at feeding (P<0.05). Although there was no overall effect of tannins on N retention in mature wether sheep, it is likely that productive ruminants with higher protein requirements would retain more N from silages containing chestnut tannins. Tannins added externally to grass silages may generate some benefits on N utilisation and environmental N excretions in sheep fed the silages.

A tangled tale of two teal : population history of the grey Anas gracilis and chestnut teal A. castanea of Australia

Two Australian species of teal (Anseriformes: Anatidae: Anas), the grey teal Anas gracilis and the chestnut teal A. castanea, are remarkable for the zero or near-zero divergence recorded between them in earlier surveys of mitochondrial DNA (mtDNA) diversity. We confirmed this result through wider geographical and population sampling as well as nucleotide sampling in the more rapidly evolving mtDNA control region. Any data set where two species share polymorphism as is the case here can be explained by a model of gene flow through hybridization on one hand or by incomplete lineage sorting on the other hand. Ideally, analysis of such shared polymorphism would simultaneously estimate the likelihood of both phenomena. To do this, we used the underlying principle of the IMa package to explore ramifications to understanding population histories of A. gracilis and A. castanea. We cannot reject that hybridization occurs between the two species but an equally or more plausible finding for their nearly zero divergence is incomplete sorting following very recent divergence between the two, probably in the mid-late Pleistocene. Our data add to studies that explore intermediate stages in the evolution of reciprocal monophyly and paraphyletic or polyphyletic relationships in mtDNA diversity among widespread Australian birds.

Unrelated helpers neither signal contributions nor suffer retribution in chestnut-crowed babblers

Alloparental care by distant/nonkin that accrue few kin-selected benefits requires direct fitness benefits to evolve. The pay-to-stay hypothesis, under which helpers contribute to alloparental care to avoid being expelled from the group by dominant individuals, offers one such explanation. Here, we investigated 2 key predictions derived from the pay-to-stay hypothesis using the chestnut-crowed babbler, Pomatostomus ruficeps, a cooperatively breeding bird where helping by distant/nonkin is common (18% of nonbreeding helpers). First, we found no indication that distant or nonkin male helpers advertised their contributions toward the primary male breeder. Helpers unrelated to both breeders were unresponsive to provisioning rates of the dominant male, whereas helpers that were related to either the breeding male or to both members of the pair were responsive. In addition, unrelated male helpers did not advertise their contributions to provisioning by disproportionately synchronizing their provisioning events with those of the primary male breeder or by provisioning nestlings immediately after him. Second, no helper, irrespective of its relatedness to the dominant breeders, received aggression when released back into the group following temporary removal for 1-2 days. We therefore find no compelling support for the hypothesis that pay-to-stay mechanisms account for the cooperative behavior of unrelated males in chestnut-crowned babblers.

Thermoanalytical study and characterization of native starches of Paraná pine seeds (Araucaria angustiofolia, Bert O. Ktze) and European chestnut seeds (Castanea sativa, Mill)

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Attack on Chestnut-bellied Euphonia nestlings by army ants

I report some observations of a Chestnut-bellied Euphonia (Euphonia pectoralis) nest in a lowland Atlantic Forest of southeastern Brazil during the early nestling period. During 7.5 hours of observations, the nest was attended 46.3% of the time, 45.6% by the female and 0.7% by the male. Unattended periods lasted 16-38 min. Parents visited the nest most of the time together at 36-59 min intervals. There were 1.06 feeding visits per nestling per hour. The two nestlings in the nest ended up preyed upon by army ants (Labidus praedator, Ecitoninae). The low height of the nest (0.8 m) may have facilitated its detection by the ants.

Horse chestnut (Aesculus hippocastanum L.) starch: Basic physico-chemical characteristics and use as thermoplastic material

Starch isolated from non-edible Aesculus hippocastanum seeds was characterized and used for preparing starch-based materials. The apparent amylose content of the isolated starch was 33.1%. The size of starch granules ranged from 0.7 to 35 pm, and correlated with the shape of granules (spherical, oval and irregular). The chain length distribution profile of amylopectin showed two peaks, at polymerization degree (DP) of 12 and 41-43. Around 53% of branch unit chains had DP in the range of 11-20. A. hippocastanum starch displayed a typical C-type pattern and the maximum decomposition temperature was 317 degrees C.Thermoplastic starch (TPS) prepared from A. hippocastanum with glycerol and processed by melt blending exhibited adequate mechanical and thermal properties. In contrast, plasticized TPS with glycerol:malic acid (1:1) showed lower thermal stability and a pasty and sticky behavior, indicating that malic acid accelerates degradation of starch during processing. (C) 2014 Elsevier Ltd. All rights reserved.

The role of human-induced fire and sweet chestnut (Castanea sativa Mill.) cultivation on the long-term landscape dynamics of the southern Swiss Alps

Changes in fire occurrence during the last decades in the southern Swiss Alps make knowledge on fire history essential to understand future evolution of the ecosystem composition and functioning. In this context, palaeoecology provides useful insights into processes operating at decadal-to-millennial time scales, such as the response of plant communities to intensified fire disturbances during periods of cultural change. We provide a high-resolution macroscopic charcoal and pollen series from Guèr, a well-dated peat sequence at mid-elevation (832 m.a.s.l.) in southern Switzerland, where the presence of local settlements is documented since the late Bronze Age and the Iron Age. Quantitative fire reconstruction shows that fire activity sharply increased from the Neolithic period (1–3 episodes/1000 year) to the late Bronze and Iron Age (7–9 episodes/1000 year), leading to extensive clearance of the former mixed deciduous forest (Alnus glutinosa, Betula, deciduous Quercus). The increase in anthropogenic pollen indicators (e.g. Cerealia-type, Plantago lanceolata) together with macroscopic charcoal suggests anthropogenic rather than climatic forcing as the main cause of the observed vegetation shift. Fire and controlled burning were extensively used during the late Roman Times and early Middle Ages to promote the introduction and establishment of chestnut (Castanea sativa) stands, which provided an important wood and food supply. Fire occurrence declined markedly (from 9 to 5–6 episodes/1000 year) during late Middle Ages because of fire suppression, biomass removal by human population, and landscape fragmentation. Land-abandonment during the last decades allowed forest to partly re-expand (mainly Alnus glutinosa, Betula) and fire frequency to increase.

Identification of a homolog of Arabidopsis DSP4 (SEX4) in chestnut: its induction and accumulation in stem amyloplasts during winter or in response to the cold_

Oligosaccharide synthesis is an important cryoprotection strategy used by woody plants during winter dormancy. At the onset of autumn, starch stored in the stem and buds is broken down in response to the shorter days and lower temperatures resulting in the buildup of oligosaccharides. Given that the enzyme DSP4 is necessary for diurnal starch degradation in Arabidopsis leaves, this study was designed to address the role of DSP4 in this seasonal process in Castanea sativa Mill. The expression pattern of the CsDSP4 gene in cells of the chestnut stem was found to parallel starch catabolism. In this organ, DSP4 protein levels started to rise at the start of autumn and elevated levels persisted until the onset of spring. In addition, exposure of chestnut plantlets to 4 °C induced the expression of the CsDSP4 gene. In dormant trees or cold-stressed plantlets, the CsDSP4 protein was immunolocalized both in the amyloplast stroma and nucleus of stem cells, whereas in the conditions of vegetative growth, immunofluorescence was only detected in the nucleus. The studies indicate a potential role for DSP4 in starch degradation and cold acclimation following low temperature exposure during activity–dormancy transition.

Evaluation of the influence of visual parameters on wave transmission velocity in sawn chestnut timber

Non-destructive, visual evaluation and mechanical testing techniques were used to assess the structural properties of 374 samples of chestnut (Castanea sativa). The principal components method was applied to establish and interpret correlations between variables obtained of modulus of elasticity, bending strength and density. The static modulus of elasticity presented higher correlation values than those obtained using non-destructive methods. Bending strength presented low correlations with the non-destructive parameters, but there was some relation to the different knot ratios defined. The relationship was stronger with the most widely used ratio, CKDR. No significant correlations were observed between any of the variables and density.

Genetic characterization of chestnut (Castanea sativa Mill.) orchards and traditional nut varieties in El Bierzo, a glacial refuge and major cultivation site in Northwestern Spain

This paper presents a detailed genetic study of Castanea sativa in El Bierzo, a major nut production region with interesting features. It is located within a glacial refuge at one extreme of the distribution area (northwest Spain); it has a centenary tradition of chestnut management; and more importantly, it shows an unusual degree of genetic isolation. Seven nuclear microsatellite markers were selected to analyze the genetic variability and structure of 169 local trees grafted for nut production. We analyzed in the same manner 62 local nuts. The selected loci were highly discriminant for the genotypes studied, giving a combined probability of identity of 6.1 × 10−6. An unprecedented density of trees was sampled for this project over the entire region, and nuts were collected representing 18 cultivars marketed by local producers. Several instances of misclassification by local growers were detected. Fixation index estimates and analysis of molecular variance (AMOVA) data are supportive of an unexpectedly high level of genetic differentiation in El Bierzo, larger than that estimated in a previous study with broader geographical scope but based on limited local sampling (Pereira-Lorenzo et al., Tree Genet Genomes 6: 701–715, 2010a). Likewise, we have determined that clonality due to grafting had been previously overestimated. In line with these observations, no significant spatial structure was found using both a model-based Bayesian procedure and Mantel’s tests. Taken together, our results evidence the need for more fine-scale genetic studies if conservation strategies are to be efficiently improved.

Molecular tools to improve chestnut management: El Bierzo as a case study

The European chestnut (Castanea sativa Mill.) is a multipurpose species that has been widely cultivated around the Mediterranean basin since ancient times. New varieties were brought to the Iberian Peninsula during the Roman Empire, which coexist since then with native populations that survived the last glaciation. The relevance of chestnut cultivation has being steadily growing since the Middle Ages, until the rural decline of the past century put a stop to this trend. Forest fires and diseases were also major factors. Chestnut cultivation is gaining momentum again due to its economic (wood, fruits) and ecologic relevance, and represents currently an important asset in many rural areas of Europe. In this Thesis we apply different molecular tools to help improve current management strategies. For this study we have chosen El Bierzo (Castile and Leon, NW Spain), which has a centenary tradition of chestnut cultivation and management, and also presents several unique features from a genetic perspective (next paragraph). Moreover, its nuts are widely appreciated in Spain and abroad for their organoleptic properties. We have focused our experimental work on two major problems faced by breeders and the industry: the lack of a fine-grained genetic characterization and the need for new strategies to control blight disease. To characterize with sufficient detail the genetic diversity and structure of El Bierzo orchards, we analyzed DNA from 169 trees grafted for nut production covering the entire region. We also analyzed 62 nuts from all traditional varieties. El Bierzo constitutes an outstanding scenario to study chestnut genetics and the influence of human management because: (i) it is located at one extreme of the distribution area; (ii) it is a major glacial refuge for the native species; (iii) it has a long tradition of human management (since Roman times, at least); and (iv) its geographical setting ensures an unusual degree of genetic isolation. Thirteen microsatellite markers provided enough informativeness and discrimination power to genotype at the individual level. Together with an unexpected level of genetic variability, we found evidence of genetic structure, with three major gene pools giving rise to the current population. High levels of genetic differentiation between groups supported this organization. Interestingly, genetic structure does not match with spatial boundaries, suggesting that the exchange of material and cultivation practices have strongly influenced natural gene flow. The microsatellite markers selected for this study were also used to classify a set of 62 samples belonging to all traditional varieties. We identified several cases of synonymies and homonymies, evidencing the need to substitute traditional classification systems with new tools for genetic profiling. Management and conservation strategies should also benefit from these tools. The avenue of high-throughput sequencing technologies, combined with the development of bioinformatics tools, have paved the way to study transcriptomes without the need for a reference genome. We took advantage of RNA sequencing and de novo assembly tools to determine the transcriptional landscape of chestnut in response to blight disease. In addition, we have selected a set of candidate genes with high potential for developing resistant varieties via genetic engineering. Our results evidenced a deep transcriptional reprogramming upon fungal infection. The plant hormones ET and JA appear to orchestrate the defensive response. Interestingly, our results also suggest a role for auxins in modulating such response. Many transcription factors were identified in this work that interact with promoters of genes involved in disease resistance. Among these genes, we have conducted a functional characterization of a two major thaumatin-like proteins (TLP) that belongs to the PR5 family. Two genes encoding chestnut cotyledon TLPs have been previously characterized, termed CsTL1 and CsTL2. We substantiate here their protective role against blight disease for the first time, including in silico, in vitro and in vivo evidence. The synergy between TLPs and other antifungal proteins, particularly endo-p-1,3-glucanases, bolsters their interest for future control strategies based on biotechnological approaches.

A cytoplasmically transmissible hypovirulence phenotype associated with mitochondrial DNA mutations in the chestnut blight fungus Cryphonectria parasitica.

Mutations causing mitochondrial defects were induced in a virulent strain of the chestnut blight fungus Cryphonectria parasitica (Murr.) Barr. Virulence on apples and chestnut trees was reduced in four of six extensively characterized mutants. Relative to the virulent progenitor, the attenuated mutants had reduced growth rates, abnormal colony morphologies, and few asexual spores, and they resembled virus-infected strains. The respiratory defects and attenuated virulence phenotypes (hypovirulence) were transmitted from two mutants to a virulent strain by hyphal contact. The infectious transmission of hypovirulence occurred independently of the transfer of nuclei, did not involve a virus, and dynamically reflects fungal diseases caused by mitochondrial mutations. In these mutants, mitochondrial mutations are further implicated in generation of the attenuated state by (i) uniparental (maternal) inheritance of the trait, (ii) presence of high levels of cyanide-insensitive mitochondrial alternative oxidase activity, (iii) cytochrome deficiencies, and (iv) structural abnormalities in the mtDNA. Hence, cytoplasmically transmissible hypovirulence phenotypes found in virus-free strains of C. parasitica from recovering trees may be caused by mutant forms of mtDNA.

EVALUATION OF COLOUR AND TEXTURE OF A PORTUGUESE TRADITIONAL SWEET: “EGG CHESTNUT”

This work aimed to evaluate the colour and texture of a Portuguese traditional sweet “egg chestnuts” along a storage period of 12 days under refrigeration. For this, the colour coordinates were measured through a colorimeter and the textural parameters were evaluated by compression and puncture tests using a textutrometer. The results showed that the colour coordinates do not vary along time, corresponding to very low values of total colour difference. Hardness, chewiness and springiness decreased in the first 3 days but after that the values were maintained approximately constant. In contrast, resilience and cohesiveness did not change along time. While crust firmness increased along storage, both the flesh firmness and stickiness did not change. The adhesiveness measured through the penetration test decreased considerably from the first to the last day. Hence, it was concluded that colour is better preserved than texture along a 12 day storage period under refrigeration.