Salivary Amylase Induction by Tannin-Enriched Diets as a Possible Countermeasure Against Tannins

Tannins are characterized by protein-binding affinity. They have astringent/bitter properties that act as deterrents, affecting diet selection. Two groups of salivary proteins, proline-rich proteins and histatins, are effective precipitators of tannin, decreasing levels of available tannins. The possibility of other salivary proteins having a co-adjuvant role on host defense mechanisms against tannins is unknown. In this work, we characterized and compared the protein profile of mice whole saliva from animals fed on three experimental diets: tannin-free diet, diet with the incorporation of 5% hydrolyzable tannins (tannic acid), or diet with 5% condensed tannins (quebracho). Protein analysis was performed by one-dimensional gel electrophoresis combined with Matrix-Assisted Laser Desorption Ionization-Time of Flight mass spectrometry to allow the dynamic study of interactions between diet and saliva. Since abundant salivary proteins obscure the purification and identification of medium and low expressed salivary proteins, we used centrifugation to obtain saliva samples free from proteins that precipitate after tannin binding. Data from Peptide Mass Fingerprinting allowed us to identify ten different proteins, some of them showing more than one isoform. Tannin-enriched diets were observed to change the salivary protein profile. One isoform of α-amylase was overexpressed with both types of tannins. Aldehyde reductase was only identified in saliva of the quebracho group. Additionally, a hypertrophy of parotid salivary gland acini was observed by histology, along with a decrease in body mass in the first 4 days of the experimental period

Comparison of Electrophoretic Protein Profiles from Sheep and Goat Parotid Saliva

Saliva provides a medium for short-term adaptation to changes in diet composition, namely, the presence of plant secondary metabolites. Salivary proteins have biological functions that have particular influence on oral homeostasis, taste, and digestive function. Some salivary proteins, such as proline-rich proteins, are present in browsers but absent in grazers. Despite the significance of salivary proteins, their expression patterns in many herbivores are unknown. We investigated the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile of parotid salivary proteins from two domesticated species, one a grazer, the sheep, Ovis aries, and the other a mixed feeder, the goat, Capra hircus, both fed on the same conventional diet. With 12.5% polyacrylamide linear gels, we observed uniform patterns of salivary proteins within the two species. In the goat profile, 21 major bands were observed, and 19 in the sheep profile. Each band was subjected to peptide mass fingerprinting for purposes of identification, allowing for 16 successful protein identifications. Marked differences were observed between the species in the region of 25–35 kDa molecular weights: one band was present in significantly different intensities; three bands were present only in goats; and one band was present only in sheep. This is the first report of a comparison of the protein salivary composition of sheep and goats and suggests that future research should be conducted to reveal a physiological function for salivary proteins related to the differences in feeding behavior of these species

NIRS potential use for the determination of natural resources quality from dehesa (acorn and grass) in Montanera system for Iberian pigs.

NIRS technology has been used as an alternative to conventional methods to determinate the content of nutrients of acorns and grass from dehesa ecosystem. Dry matter (DM), crude fat (CF), crude protein (CP), starch, total phenolic compounds (TP), α-tocopherol, γ-tocopherol, fatty acids, neutral detergent fiber (NDF), total antioxidant activity (TAA) and total energy (TE) were determined by conventional methods for later development of NIRS predictive equations. The NIR spectrum of each sample was collected and for all studied parameters, a predictive model was obtained and external validated. Good prediction equations were obtained for moisture, crude fat, crude protein, total energy and γ-tocopherol in acorns samples, with high coefficients of correlation (1-VR) and low standard error of prediction (SEP) (1-VR=0.81, SEP=2.62; 1-VR=0.92, SEP=0.54; 1-VR=0.86, SEP=0.47; 1-VR=0.84, SEP=0.2; 1-VR=0.88, SEP=5.4, respectively) and crude protein, NDF, α-tocopherol and linolenic acid content in grass samples (1-VR=0.9, SEP=1.99; 1-VR=0.87, SEP=4.13; 1-VR=0.76, SEP=10.9; 1-VR=0.82, SEP=0.6, respectively). Therefore, these prediction models could be used to determinate the nutritional composition of Montanera natural resources

Assessing foraging strategies of herbivores in Mediterranean oak woodlands: a review of key issues and selected methodologies.

Montados are agro-silvo-pastoral ecosystems, typical of the Southwest Iberian Peninsula, of high socio-economic and conservation importance,where grazing is a dominant activity. Montados are characterized by na open tree canopy of Quercus sp. and a diverse undercover of shrubs and grasslands that constitute the plant food resources for grazing animals. Plant food resources of Montados are highly variable, both spatially and seasonally, in quantity and quality. Reliable and easy to use methods to monitor grazing are necessary to allow proper understanding of foraging strategies of grazing animals and to set sustainable grazing management. We describe the main characteristics of the plant food resources available for grazing animals, striking its variability, and revise the potential of using N-alkanes and saliva

Sheep and goat saliva proteome analysis: A useful tool for ingestive behavior research?

Sheep and goats differ in diet selection, which may reflect different abilities to deal with the ingestion of plant secondary metabolites. Although saliva provides a basis for immediate oral information via sensory cues and also a mechanism for detoxification, our understanding of the role of saliva in the pre-gastric control of the intake of herbivores is rudimentary. Salivary proteins have important biological functions, but despite their significance, their expression patterns in sheep and goats have been little studied. Protein separation techniques coupled to mass spectrometry based techniques have been used to obtain an extensive comprehension of human saliva protein composition but far fewer studies have been undertaken on animals' saliva. We used two-dimensional electrophoresis gel analysis to compare sheep and goats parotid saliva proteome. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to identify proteins. From a total of 260 sheep and 205 goat saliva protein spots, 117 and 106 were identified, respectively. A high proportion of serum proteins were found in both salivary protein profiles. Major differences between the two species were detected for proteins within the range of 25–35 kDa. This study presents the parotid saliva proteome of sheep and goat and highlights the potential of proteomics for investigation relating to intake behavior research

Effect of condensed tannin ingestion in sheep and goat parotid saliva proteome

Saliva appears as a defence mechanism, against potential negative effects of tannins, in some species of animals which have to deal with these plant secondary metabolites in their regular diets. This study was carried out to investigate changes in parotid saliva protein profiles of sheep Ovis aries)and goats (Capra hircus), induced by condensed tannin ingestion. Five Merino sheep and five Serpentina goats were maintained on a quebracho tannin enriched diet for 10 days. Saliva was collected through catheters inserted on parotid ducts and salivary proteins were separated by two dimensional gel electrophoresis. Matrix assisted Laser desorption ionization – time of flight(MALDI-TOF)and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to identify the proteins whose expression levels changed after tannin consumption. Although no new proteins appeared, quebracho tannin consumption increased saliva total protein concentration and produced changes in the proteome of both species. While some proteins were similarly altered in both species parotid salivary protein profile, sheep and goats also presented species-specific differences in response to tannin consumption

Progress in analytical approaches integrating Livestock and Biodiversity to identify HNV Montados

The World Congress Silvo-Pastoral Systems 2016 aims to gather researchers from different disciplines, practitioners and policy makers at different governance levels that deal with the management and sustainability of silvo-pastoral systems. In this way the congress will create a fertile context to progress through interdisciplinarity research approaches that can help translate scientific knowledge into new adaptive management solutions, and thus bridge from science to practice. The aim is also to gather and compare knowledge from silvo-pastoral systems around the world, which share drought as a limiting factor, so that they can be discussed and evaluated

Progress in Identifying High Nature Value Montados: Impacts of Grazing on Hardwood Rangeland Biodiversity

Due to their complex structure and traditional low-intensity management, Portuguese oak woodland rangelands known as montados are often considered high nature value (HNV) farming systems, and as such, they may be deemed eligible for subsidies and incentives by governmental and nongovernmental agencies. Too little is known about how the HNV concept might be applied to conserve complex silvopastoral systems. These systems, due to their structural and functional complexity at multiple scales, tend to support high levels of biodiversity. Montados are in sharp decline as a result of the rapid specialization of land management that, through simplification, undermines multifunctionality. Understanding how changes in management influence these systems and their biodiversity is needed for prioritizing conservation efforts and for ensuring they remain HNV systems. On the basis of a field survey in 58 plots distributed among 29 paddocks on 17 farms, we conducted an integrated analysis of the relationship between grazing intensity and biodiversity in montados of similar biophysical and structural characteristics. Data on management were obtained through interviews, and biodiversity data (vegetation, macrofungi, birds, herpetofauna) were obtained through specific field protocols. Additional spatial data, such as soil characteristics, slope, land cover, and linear landscape elements, were also analyzed. The results show no overall biodiversity variation as a result of different management practices. However, different groups of species react differently to specific management practices, and within a pasture, grazing impacts are heterogenous. In low grazing intensity plots, macrofungi species richness was found to be higher, while bird species richness was lower. Using tree regeneration as proxy for montado sustainability, results show less tree regeneration in areas with higher forage quality and more intense grazing. Pathways for future progress are proposed, including creating areas within a paddock that attract grazing away from where regeneration is desired