Giovanna Piluzza

Correlations between phenolic content and antioxidant properties in twenty-four plant species of traditional ethnoveterinary use in the Mediterranean area

Context: Scientific information on antioxidant properties and phenolic content of less widely used plants can be useful. Therefore, the assessment of such properties remains an interesting and useful task, particularly for finding new sources for natural antioxidants, functional foods, and nutraceuticals. Objective: As knowledge about antioxidant properties and phenolic content of many plant species used as traditional plant remedies is limited, we determined in vitro the total antioxidant activity and the phenolic content of several plant species traditionally used for ethnoveterinary practices. Materials and methods: For 24 extracts (70% acetone) from wild and cultivated plant species traditionally used for health care of animals we determined the Trolox equivalent antioxidant capacity (TEAC) by the two assays 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the 2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS). The phenolic content was determined by the Folin Ciocalteu method. Results: Total phenolics, calculated as gallic acid equivalent (GAE), showed variation ranging from 3.18 (Allium sativum L. (Liliaceae)) to 147.68 (Pistacia lentiscus L. (Anacardiaceae)) mgGAE/g dry weight (DW). High TEAC values corresponded to high phenolic content, while plants with low antioxidant activity exhibited low total phenolic content. The TEAC determined through each assay and total phenolic content were positively correlated, R2 = 0.9152 and R2 = 0.8896, respectively, for DPPH and ABTS assay. Discussion and conclusion: These findings suggest that phenolic content could be used as an indicator of antioxidant properties. The results of this study encourage investigations on Mediterranean plant species as sources of antioxidants.

The dynamics of phenolic concentration in some pasture species and implications for animal husbandry

BACKGROUND Although secondary metabolites have often been studied considering their primary role as plant-defensive and their effect on consumers detrimental, their effects are usually dose-dependent, so it is important to consider their content at all phenological phases and in different plant organs. In this study the content of total phenolics (TotP), non-tannic phenolics (NTP) and proanthocyanidins (PA) was determined in natural populations of pasture species at defined phenological phases and in different plant organs and also in pathogen-infected plants. RESULTS Variation in TotP, NTP and PA content was observed in all species, phenological phases and plant organs. Leaf blades were the organs with the highest content in all species. The content of all examined compounds increased in infected leaves, reaching the maximum when 20% of plant tissues were attacked and decreasing at higher infection levels. CONCLUSION Delphinidin was found as an appropriate standard related to the PA structure of each plant species. PA concentrations were always in the range considered beneficial for animals, not exceeding 60 g delphinidin equivalent kg(-1) dry matter. The Folin-Ciocalteu and butanol/HCl/Fe(3+) assays remain the most practical methods for screening large numbers of different plant species for TotP and PA.

Relationships between pasture legumes, rhizobacteria and nodule bacteria in heavy metal polluted mine waste of SW Sardinia

Local populations of the pasture legumes Astragalus hamosus, Lotus edulis, Lotus ornithopodioides, Medicago ciliaris and Scorpiurus muricatus from heavy metal polluted and unpolluted sites in Sardinia were compared for tolerance to Zn, Cd and Pb in hydroponics. Tolerance of plants to heavy metals varied significantly depending on the species, origin of the population and metal. The species L. edulis, L. ornithopodioides and M. ciliaris possessed higher metal tolerance and were used in a pot experiment with Zn, Cd and Pb polluted mine waste. Seeds were inoculated with the metal tolerant plant growth-promoting rhizobacterium Variovorax paradoxus 5C-2 or/and with the corresponding symbiotic nodule bacteria containing the enzyme 1-aminocyclopropane-1-carboxylate deaminase. Co-inoculation with the bacteria had synergistic and additive effects on nodule number, root growth and uptake of elements (N, P, Ca, Mg, Na, Mn, Zn and Pb) in shoots of L. edulis and L. ornithopodioides. Shoot biomass and uptake of K, Fe and Cd was increased by a combined inoculation of L. edulis. The ratio between shoot and root contents of Pb in L. ornithopodioides was above 1, suggesting a characteristic trait of hyperaccumulating species. The results suggest that the development of metal tolerant and efficient plant-bacteria systems might be useful for phytostabilization and revegetation of mine wastes.

Uses of plants, animal and mineral substances in Mediterranean ethno-veterinary practices for the care of small ruminants.

ETHNO-PHARMACOLOGICAL RELEVANCE The cultural heritage of Sardinian shepherds is rapidly vanishing and survives in the memory of elderly people. The objective of our study was not only to report the usage of plants and their preparation for administration but also the use of other remedies of different origin arising from traditional ethno-veterinary knowledge, as Sardinian shepherds were used to employ plants, animals, minerals and combinations of several substances to prepare remedies for prophylaxis or therapy on their animals. MATERIALS AND METHODS The work was carried out in rural areas of the island of Sardinia (Italy) by interviewing shepherds and filling questionnaires in order to record ethno-veterinary practices traditionally used for animal health care. RESULTS Ethno-veterinary remedies traditionally utilised for treatments of small ruminants against ecto-and endo-parasites, gastrointestinal diseases, viral and bacterial diseases, wounds, sprains and bruises were identified. Non herbal remedies outnumbered the herbal ones, as usually plant species were mainly used for the care of cattle and equines. A total of 150 ethno-veterinary uses were documented for the treatment of 33 animal conditions, a detailed account of the formulations and their administration to sheep and goats was provided. Herbal remedies involved the use of twenty two spontaneous species and seven cultivated species. CONCLUSIONS This study identifies remedies used in ethno-veterinary practices for small ruminants care in Sardinia, the second major Mediterranean island which has agro-pastoral activities dating back to Neolithic. Moreover, the danger of losing oral traditions, and the increasing attention towards traditional remedies as potential sources of natural products for improving animal health and welfare, support the interest of our survey.

Condensed tannin accumulation and nitrogen fixation potential of Onobrychis viciifolia Scop. grown in a Mediterranean environment

BACKGROUND Sainfoin (Onobrychis viciifolia Scop.) is a forage legume found in temperate areas but is less widespread in Mediterranean environments. Compared with other perennial legumes, it has the advantage of containing condensed tannins (CT) that can be important for their implications on ruminant nutrition and health. Data on nitrogen (N) fixation by sainfoin in the literature originate from very different environments and only a few field data are available, so it is important to improve knowledge on the N fixation potential of this species, particularly under a Mediterranean climate. Here the accumulation pattern of polyphenolic compounds (total, non-tannic polyphenols and CT) and the N fixation potential of sainfoin were studied in order to contribute to its valorisation for sustainable farming management in Mediterranean environments. RESULTS CT concentrations were always in the range considered beneficial for animals, not exceeding 50 g delphinidin equivalent kg⁻¹ dry matter (DM). The regression of aerial fixed N on aerial DM showed a relationship of 22 kg fixed N t⁻¹ aerial DM in a Mediterranean environment. CONCLUSION A wider exploitation of sainfoin is suggested for production under rain-fed conditions, thus enlarging the limited set of available perennial legumes suitable for Mediterranean environments.

Characterization of Sardinian cork oak (Quercus suber L.) genetic resources for economically important traits

Considering the very high socio economic value of cork oak (Quercus suber L.) in Mediterranean areas, a better knowledge of local genetic resources is necessary in order to reach a good balance between the conservation issues and the need of an efficient cork production network. In such frame, local germplasm of Q. suber from the main cork production areas of Sardinia was analysed for DNA and isozyme polymorphisms and for cork quality in relation to some environmental parameters. A total number of 24 sample stands of cork oak were selected, representing typical vegetation, sylvicultural and cork quality features within the eight identified cork growing areas in Sardinia. Considerable variation was found between cork oak stands of the different areas. Results of the principal component analysis performed on the cork quality data and environmental characters of the eight areas, showed that the first three components explain 72.2% of the variation. The major characters involved in this differentiation were cork quality characters such as dimensional recover, moisture and Mg content of the cork and also elevation of stands m a.s.l. for the first component. The second component appears to be determined by some climatic parameters (average annual temperature and average of the minimum temperatures of coldest month) and by the Fe and Zn in the cork. For the third component, major characters involved in differentiation are average annual rainfall, bark thickness and Ca content in the cork.

Cell-based and chemical assays of the ability to modulate the production of intracellular Reactive Oxygen Species of eleven Mediterranean plant species related to ethnobotanic traditions

The aim of this study was to compare three tests for the evaluation of the antioxidant potential of eleven plant species traditionally used to cure or to enhance animal and/or human wellbeing. In addition to the ability to modulate the production of intracellular reactive oxygen species the effect of the treatment on cell viability was also considered. In the cell-based experiment DCF (2′,7′dichlorofluorescein diacetate) and in the chemical assays DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt), Pistacia lentiscus exhibited high antioxidant capacity, while Cistus creticus and Euphorbia characias were ineffective in cell-culture and displayed high activity in the chemical assays, and others (Malva sylvestris, Matricaria chamomilla, Urtica dioica) showed an opposite trend. Olea europaea (sylvestris and europaea) and Umbilicus rupestris displayed highest antioxidant efficacy in both HUVEC and HL-60 cells, and provided also appreciable activity in the chemical assays. The study gives evidence that herbal extracts may have similar antioxidant properties but different effects on cells. Therefore, it is important to combine analytical and biological testing methods in order to measure the antioxidant capacity of natural extracts. Results encourage to explore further the potential nutraceutical and functional properties of Mediterranean plant species related to ethno-botanic traditions for animal health care.

Traditional knowledge about plant, animal, and mineral-based remedies to treat cattle, pigs, horses, and other domestic animals in the Mediterranean island of Sardinia

BackgroundMediterranean farmers traditionally utilized plants, animals, and minerals sourced locally to treat their animals. Research is needed to understand at what extent such knowledge of domestic animal care still survives and to document such traditions for further developments.MethodsWe carried out our field study to recover ancient ethno-veterinary practices by means of questionnaires and interviews to farmers in rural areas of the Mediterranean island of Sardinia (Italy). Quantitative indices were used to evaluate the distribution and diversity of the acquired information.ResultsWe report here 98 sources (42 plant taxa, 14 animal-based substances, 15 minerals, and 27 other materials of various origin) emerged from the survey for the care of 41 ailments of cattle, pigs, and horses. Ethno-veterinary treatments, detailed in their formulations and applications, were used against ecto- and endo-parasites, gastrointestinal diseases, heart diseases, viral and bacterial diseases, wounds, sprains, and bruises.ConclusionOur survey can be useful to implement the use of phyto-therapeutics and other remedies of non-herbal origin for diseased animals, and, as elderly farmers held most of the knowledge, it can contribute to the conservation of Mediterranean ethno-veterinary knowledge.

Bioactive compounds and antioxidants from a Mediterranean garland harvested at two stages of maturity

Abstract Chrysanthemum coronarium L. (garland) is an herbaceous plant rich in bioactive compounds. The chemical composition, bioactive compounds and antioxidant properties of a Mediterranean garland population were investigated in different organs at two phenological stages. Antioxidant capacity varied from 7.9 (vegetative) to 14.4 (flowering) mmol Trolox equivalent antioxidant capacity 100 g−1 dry weight (DW). A significant correlation between antioxidant capacity and total phenolics and total flavonoids was found at flowering stage. LC-MS/MS analysis revealed that chlorogenic acid reached a maximum of 4.7 μg g−1 DW in leaves; flowers were high in luteolin (2.37 μg g−1 DW), whereas leaves showed a remarkable content of rutin (1.78 μg g−1 DW). Results highlight differences in bioactive compound levels and antioxidant capacity related to plant stages and organs. This research provides new insights into antioxidant activities and chemistry of garland, in view of its exploitation in areas of fodder resources, functional foods and natural antioxidants.

Cropping systems sustainability: Inoculation and fertilisation effect on sulla performances in a new cultivation area

To assess the feasibility of the sulla [Sulla coronaria (L.) Medik] forage legume in a new agroecosystem, its host-specific symbiotic interaction needs to be taken into account. This study aimed to investigate the effects of inoculation and nitrogen (N) fertiliser on productive performances and N-fixation ability of sulla established in a new habitat within a Mediterranean agropastoral area. Sulla plants, previously inoculated (with peat-based, liquid inoculants, and using soil from an existing sulla field) and unfertilised or N fertilised were evaluated in Sardinia (Italy). During 2013-2014, sulla plants were sampled at four growing stages, from vegetative stage to seed set, and shoot length, shoot dry matter (DM) yield and N content were monitored. Moreover, atom% 15N isotopic excess, proportion of N derived from the atmosphere and fixed N of sulla shoots were quantified. Inoculation and N fertilisation both affected growth, DM and N yields, and N-fixation of sulla. Compared to the best inoculated treatment, the DM yield and fixed N of the control only represented 10 to 22% and 2 to 11%, respectively. Nitrogen fertilisation caused temporary decreases in the N fixing ability of sulla. Results pointed out that rhizobial inoculation is essential for the exploitation of sulla outside its traditional cropping area.