In Vitro Anthelmintic Activity of Sea Buckthorn (Hippophae rhamnoides) Berry Juice against Gastrointestinal Nematodes of Small Ruminants

Simple Summary Gastrointestinal nematodes are included worldwide among the most prominent parasites of small ruminants. In past decades, the control of these nematodes mainly relied on the use of synthetic anthelmintic drugs. However, nowadays the exclusive use of anthelmintic drugs is considered an obsolete and unsustainable control strategy due to the onset of anthelmintic-resistant sheep gastrointestinal nematode strains and the issues linked with the environmental pollution and residues in food of animal origin of synthetic anthelmintic drugs. Among alternative or complementary methods, the use of plants endowed with anthelmintic properties has been identified as a valuable option. As a part of this approach, this study evaluated the in vitro anthelmintic properties of two commercial sea buckthorn (Hippophae rhamnoides) berry juices on sheep gastrointestinal nematodes. Both H. rhamnoides berry juices tested in this study showed interesting anthelmintic properties in vitro. The obtained results are promising regarding the use of sea buckthorn berry juice as a potential tool for the control of gastrointestinal nematodes in small ruminants. Gastrointestinal nematodes are one of the major threats in small ruminant breeding. Their control is difficult due to the development of anthelmintic resistance, and the search for new molecules endowed with anthelmintic activity (AH) is considered a priority. In this context, we evaluated the in vitro AH activity of two commercial sea buckthorn (Hippophae rhamnoides) berry juices, namely SBT and SBF. The in vitro evaluation was based on the egg-hatch test and larval exsheathment assay at different concentrations. Data were statistically analysed, and the EC50 was calculated. Chemical analyses were performed to evaluate the total polyphenol content of the juices and chemical profile of the most represented compounds. The role of the polyphenolic fraction in the anthelmintic activity of the juices was also assessed. At the highest concentrations, the activity of SBT was high in both tests and comparable to that observed in the thiabendazole-treated positive controls, while SBF showed a lower efficacy. Glycosylated isorhamnetin and quercetin were the most represented polyphenolic compounds in both juices. In conclusion, both H. rhamnoides berry juices tested in this study showed interesting anthelmintic properties in vitro

Characterization of the endophytic bacterial community of Bituminaria bituminosa plant grown in vitro and its interaction with the plant extract

IntroductionBituminaria bituminosa is a medicinal plant recognized for its phytochemicals, such as furanocoumarins, pterocarpans, and flavonoids. Since the secondary metabolism is influenced by the plant-endophyte interactions, the endophytic bacterial community of B. bituminosa was explored and the possible interactions with the plant were described.Materials and methodsDifferent bacterial strains were isolated from different organs of in vitro plants as shoots, roots, and seeds. The bacterial strains were identified and phenotypically characterized for different traits; strains were also exposed to different concentrations of B. bituminosa plant extract showing different susceptibility, probably determined by different secondary metabolites produced by the plant in the different organs (i.e. aerial parts and roots).Results and discussionBacterial strains showed different phenotypic characteristics; the 6 detected haplotypes were dominated by a single species related to Stenotrophomonas rhizophila. Endophytes isolated from the aerial parts produced a higher indole-3-acetic acid (IAA) amount than those of the roots, while all strains were unable to produce biosurfactants and antagonistic activity toward the other strains. The research opens new perspectives for future analysis addressed to test the susceptibility of the endophytic bacterial community of B. bituminosa toward the pure compounds extracted from the plants, and to investigate the role of these compounds on the distribution of endophytes within the different plant tissues

Do changes in Lactuca sativa metabolic performance, induced by mycorrhizal symbionts and leaf UV-B irradiation, play a role towards tolerance to a polyphagous insect pest?

: The increased ultraviolet radiation (UV) due to the altered stratospheric ozone leads to multiple plant physiological and biochemical adaptations, likely affecting their interaction with other organisms, such as pests and pathogens. Arbuscular mycorrhizal fungi (AMF) and UV-B treatment can be used as eco-friendly techniques to protect crops from pests by activating plant mechanisms of resistance. In this study, we investigated plant (Lactuca sativa) response to UV-B exposure and Funneliformis mosseae (IMA1) inoculation as well as the role of a major insect pest, Spodoptera littoralis. Lettuce plants exposed to UV-B were heavier and taller than non-irradiated ones. A considerable enrichment in phenolic, flavonoid, anthocyanin, and carotenoid contents and antioxidant capacity, along with redder and more homogenous leaf color, were also observed in UV-B-treated but not in AMF-inoculated plants. Biometric and biochemical data did not differ between AMF and non-AMF plants. AMF-inoculated plants showed hyphae, arbuscules, vesicles, and spores in their roots. AMF colonization levels were not affected by UV-B irradiation. No changes in S. littoralis-feeding behavior towards treated and untreated plants were observed, suggesting the ability of this generalist herbivore to overcome the plant chemical defenses boosted by UV-B exposure. The results of this multi-factorial study shed light on how polyphagous insect pests can cope with multiple plant physiological and biochemical adaptations following biotic and abiotic preconditioning

Celiachia e Sport: valutazione delle problematiche e degli strumenti disponibili.

Le diagnosi di celiachia in Italia e nel mondo, negli ultimi anni, sono aumentate significativamente: ciò non significa necessariamente che la celiachia sia in crescita ma molto probabilmente che, con l’aumento delle conoscenze e della sensibilità tra gli operatori sanitari, migliorano le possibilità diagnostiche della malattia. Anche gli atleti professionisti e gli sportivi, in genere, non sfuggono alla problematica e i preparatori atletici, gli allenatori, i team di medicina dello sport e tutti i professionisti che lavorano in tale ambito devono confrontarsi con una malattia che può presentarsi in maniera insidiosa, compromettendo notevolmente le prestazioni atletiche, e che richiede una adeguata gestione dell’alimentazione, al momento unica terapia della malattia. Il presente lavoro è stato articolato in diverse parti. Nella prima parte è stata effettuata un’approfondita ricerca bibliografica, riguardante i diversi aspetti connessi con la malattia celiaca, la dieta senza glutine e l’alimentazione del soggetto sportivo, in modo tale da poter comprendere le possibili problematiche, sia nutrizionali che pratiche, relative all’alimentazione gluten-free, in soggetti con tale patologia, che svolgono attività fisica di resistenza a moderata o elevata intensità. Nella seconda parte del lavoro è stato sviluppato un questionario, formulato sulla base delle conoscenze acquisite durante la prima parte del lavoro, che poi è stato somministrato a soggetti con i suddetti requisiti, con lo scopo di indagare le difficoltà effettivamente riscontrate. Successivamente, stati delineati i possibili strumenti a disposizione dei soggetti celiaci che potrebbero essere utilizzati per superare le problematiche riscontrate, con una maggiore attenzione nei riguardi del soggetto sportivo. Infine, nell’ultima parte del lavoro, è stato sviluppato un piano nutrizionale basato su due giornate tipo di un atleta aerobico, utilizzando in una versione cereali contenenti glutine e nell’altra cereali privi di glutine, in modo tale da andare a valutare le possibili carenze e gli sbilanciamenti nutrizionali a cui i soggetti sportivi celiaci possono andare in contro. La nutrizione dell’atleta celiaco presenta indubbiamente maggiori complessità rispetto all’atleta sano, ma se l’intolleranza al glutine viene gestita correttamente, non determina limitazioni nell’attività sportiva e nel raggiungimento della massima prestazione. 2. ABSTRACT In the last decades, the diagnosis of celiac disease, in Italy and worldwide, has increased significantly. This raise is related both to an effective increase of the cases and to an improvement of the diagnostic possibilities of the disease. The rising number of diagnosis necessarily leads to an increasing frequency of celiac disease even in professional athletes and athletes in general. So athletic trainers, coaches, the sports medicine team and other professionals have to face daily this disorder, that can be insidious and that requires a strictly dietary management. The present study includes several steps. In the first part, a literature research was carried out and it concerns many aspects related to celiac disease, gluten free diet, celiac disease in sports, nutrition of celiac athletes and difficulties and risk for these people. In the next part of the study a questionnaire was developed on the bases of the knowledges acquired in the previous step of the work and administered to several celiac athletes. The purpose of the survey was to determine both nutritional and practical risks and issues, in order to outline strategies and tools that can help these people to manage the association between sport and disease. In the last part of the study a nutritional plan based on two typical days of a resistance athletes was developed, first using aliments containing gluten and after aliments without gluten. The purpose was to investigate if gluten free diet can lead to nutritional deficiency and imbalances. The nutrition of celiac athletes several critical points, but if it is correctly managed, it doesn’t determine limitation in sports

Comparison of the Chemical and Sensorial Evaluation of Dark Chocolate Bars

As it mimics olfactory perception, headspace analysis is frequently used for examination of products like chocolate, in which aroma is a key feature. Chemical analysis by itself, however, only provides half the picture, as final consumer’s perception cannot be compared to that of a Gas Chromatography-Mass Spectrometry (GC-MS) port, but rather to a panel test assessment. The aim of the present study was the evaluation of combined chemical (by means of headspace solid-phase microextraction and GC-MS) and panel test data (by means of a sensory evaluation operated by 6 untrained panelists) obtained for 24 dark chocolate bars to assess whether these can discriminate between bars from different brands belonging to different commercial segments (hard discount, HD; supermarket, SM; organic bars, BIO). In all samples, with the only exception of one supermarket bar (in which esters exhibited the highest relative abundance), pyrazines were detected as the most abundant chemical class (HD: 56.3–74.2%; BIO: 52.0–76.4%; SM: 31.2–88.9%). Non-terpene alcohols, aldehydes, and esters followed as quantitatively relevant groups of compounds. The obtained data was then subjected to hierarchical cluster (HCA) and principal component (PCA) analysis. The statistical distribution of samples obtained for the chemical data did not match that obtained with panelists’ sensorial data. Moreover, although an overall ability of grouping samples of the same commercial origin was evidenced for hard discount and supermarket bars, no sharp grouping was possible

Plant volatiles in the agri-food sector with particular regard to Cannabis sativa L. and wine traceability

Volatile Organic Compounds emitted by plants are a mixture of lipophilic compounds that exert a role in plant communication with other organisms as well as in plant defence against unfavourable environmental conditions. Due to their roles, the analysis of both spontaneously emitted volatiles and essential oil, may represent an important tool to be exploited in various important sectors, including agricultural and food ones, since it can provide a series of information on plant behaviour. The first section of the dissertation consists of a brief introduction on plant volatiles, both spontaneously emitted and EO, as they represent a recurrent theme of the thesis. In the second chapter, variations in the volatile emission as a response to environmental factors were evaluated. Indeed, the former study investigated the potential impacts of the UV-B exposition and the inoculation with the mycorrhizal fungi Funneliformis mosseae on the volatile chemical profile of Lactuca sativa L., an important annual herb widely cultivated as leaf vegetables. Results highlighted UV-B exposure as an important abiotic factor capable of influencing plant secondary metabolism. Conversely, mycorrhization, which could not be considered a biotic factor, but rather a mutualistic symbiosis, was not able to activate changes in the biosynthetic pathways involved in the volatile compound production in lettuce plants. The following two studies of this chapter evaluated the effect of narrowband LED light supplementation to ambient light on basil volatile metabolism. The former evaluated the chemical composition of the spontaneous volatile emission of two different varieties of sweet basil (Tigullio, TI and Red Rubin, RR) while the latter the variations of the chemical composition and hydrodistillation yield of the essential oil obtained only from Tigullio. Concerning volatile compounds spontaneously emitted from basil varieties, monochromatic wavebands have been shown to stimulate changes in the metabolic pathways, mainly in green-leafed basil, but also in red-leafed ones. In fact, the spontaneous volatile emission of basil plants was influenced by the light treatments, suggesting a practical application for greenhouse cultivation since it was able to confer a better pleasantness to the Tigullio cultivar, increasing the linalool content and reducing the biosynthesis of eugenol, which is considered one of the major unpleasant compounds in sweet basil. LED light supplementation was also able to affect the chemical composition of the essential oil obtained from O. basilicum cv. Tigullio. In particular, blue and green lights were more efficient in enhancing the biosynthesis of sesqui- and diterpenes, while red and white lights determined an increase of oxygenated monoterpenes, mainly of linalool and 1,8-cineole. Since basil plants producing good amounts of linalool are characterised by the highest consumer acceptability, the present study suggests, once again, a practical application of light modulation for greenhouse cultivation. The fourth study of this section aimed to evaluate the presence of a pattern relating the chemical composition of EOs and volatile organic compounds spontaneously emitted by flowers and leaves of 12 species of Salvia L. to their geographical origin, as well as to their clade distribution. Therefore, since both the EOs and the spontaneous volatile emission obtained from flowers and leaves were evaluated, we were able to highlight which analysis was more suitable to show this correlation. A great variability was found in the chemical volatile profiles of the analysed Salvia species, which, however, seems to be strictly related to their genetic heritage and, in turn, to their geographical origin. The multivariate statistical analyses, indeed, evidenced a slight correspondence between the EO chemical compositions and both the geographical origin and the clade distribution of Salvia species, while the same trend was not found for both the flowers and the leaves emissions. Given that all the species analysed in the present study were cultivated under the same environmental conditions, the similarities and differences shown in the volatile profiles could be considered genetically determined. Finally, the last study of the second chapter deals with the investigation of the spontaneous volatile emissions of inflorescences and leaves of Olea europaea accessions, collected in different municipal areas of Toscolano Maderno territory and belonging to different autochthon cultivars, in order to evaluate the influence of cultivar, environment, and their interaction on the plant volatile profile. The interaction between cultivar and environment has proved to be the most effective factor able to affect the volatile emissions of both inflorescences and leaves, which, instead, were not significantly influenced by single factors. Multivariate statistical treatments, however, evidenced a tendency for the samples to gather according to the area of provenance rather than by cultivar, and a better partitioning was obtained by considering the volatile emissions of the inflorescences. The third chapter of this PhD thesis aims to increase the knowledge of Cannabis sativa L. EO, a valuable by-product obtained from hemp inflorescences, which are usually discarded by the main supply chains of this crop. Hemp EO has attracted increasing interest thanks to its various applications in different fields. However, its chemical composition and productivity seem to be influenced by various environmental and meteorological conditions and these factors, besides the lack of standardisation of growing and operating conditions, make the comparison among the data reported in diverse studies quite challenging. Thus, the objective of the first study of this section was to investigate the chemical composition and the yield of the EO obtained from eleven industrial genotypes of C. sativa, cultivated and collected for two consecutive growing seasons, under the same cultural conditions. The results of the present study showed a significant influence of both the genotype and the year of cultivation, as well as of their interaction on the EO chemical profiles and yields. The second study of this chapter, instead, aimed to evaluate the effect of the phenological development on the production of the EO obtained from the inflorescences of two genotypes that in the previous study gave interesting results: Codimono (monoecious) and Fibrante (dioecious). Terpenes and cannabinoids represented the major detected chemical classes. Among the former class, sesquiterpene hydrocarbons were the prevalent components in the EOs of all phenological stages even though they showed a tendency to decrease during the plant phenological development. Conversely, oxygenated sesquiterpenes and phytocannabinoids showed a trend to increase during the plant growth, likewise the EO productivity. Therefore, despite its variability, hemp EO is consistently characterized by terpenes and phytocannabinoids and, among terpenes, sesquiterpenes are the most variable components, both in their hydrocarbon and oxygenated forms. Taking this into account, the third study of this section deals with the evaluation of the ability of the EOs obtained from two industrial genotypes of C. sativa, characterized by different chemical compositions, to counteract the microglia-mediated neuroinflammation using a human microglia cell model (HMC3), and both the volatile oils exhibited good outcomes in terms of anti-inflammatory activity. Finally, the last work of chapter 3, aimed to investigate the in vitro inhibitory activity of the essential oils obtained from leaves and inflorescences of two genotypes of C. sativa L. against 21 antibiotic-resistant field strains of Staphylococcus pseudintermedius, a bacterium responsible for the canine pyoderma and otitis externa. Finally, the fourth chapter reported different applications of volatile analysis and other analytical investigations in the food sector. A particular focus was dedicated to the traceability and authenticity of wine, which represent important factors for consumers’ and producers’ protection. Therefore, the aims of the first two studies of this section were the evaluation of volatile and multi-elemental analysis for the traceability of Italian wines coming from different regions. Volatile organic compounds, constituting important wine features able to influence consumer acceptability, have been reported by the literature as an important tool for wine authentication. In the present study, different chemometric approaches were applied to evaluate the ability of volatile fingerprints to discriminate Italian wines according to their region of origin. Multivariate statistical methods, in particular PCA and HCA, exhibited a primary separation of the wines according to their type (red and white) and a good separation of red wines according to their geographical provenance. Conversely, LDA did not return successful outcomes. Inorganic elements due to their greater stability have been widely used in the food industry for tracing the wine origin. Indeed, the preliminary results of the second work of this chapter evidenced the ability of multi-elemental analysis associated with supervised statistical methods to discriminate Italian wines according to their geographical origin. This study represented a starting point for future investigations involving an increased number of samples from each region, necessary to confirm the discriminating power of the proposed method. Volatile analysis associated with other phytochemical and sensorial analyses was also applied for the evaluation of the Hibiscus rosa-sinensis-flavoured beer and Citrus-based products discussed in the present chapter. Finally, in the last work of this dissertation, the EOs obtained from different Salvia L. species were investigated for their antimicrobial activity against Listeria monocytogenes, an important gram-positive bacterium able to contaminate food products causing human listeriosis. The obtained encouraging results suggest possible applications of the analysed EOs in the food industry in different delivery systems that can enhance the bio-efficacy of the EOs while mitigating the challenge posed by their strong aroma

Halophyte Artemisia caerulescens L.: Metabolites from In Vitro Shoots and Wild Plants

Halophyte plants are potential resources to deal with the increasing soil salinity determined by climatic change. In this context, the present study aimed to investigate the germplasm conservation of Artemisia caerulescens collected in the San Rossore Estate (Pisa, Italy) through in vitro culture, biochemical properties, and the phytochemical composition of the volatile fraction of both in vitro shoots and different organs of wild plants (leaves, young and ripe inflorescences). The best medium tested for the shoot proliferation was MS, with the addition of 1 μM BA. Total polyphenol content and antioxidant activity were noticeable in both the inflorescences, while leaves and in vitro shoots showed lower amounts. Concerning the phytochemical investigation, the headspaces (HSs) and the essential oils (EOs) were characterized by oxygenated monoterpenes as the main chemical class of compounds in all samples, and with α- and β-thujone as the major constituents. However, the EOs were characterized by noticeable percentages of phenylpropanoids (23.6–28.8%), with brevifolin as the unique compound, which was not detected in the spontaneous volatile emissions of the same parts of the wild plant. Good amounts of EOs were obtained from different organs of the wild plant, comprising between 0.17% and 0.41% of the young and ripe inflorescences, respectively

Halophyte <i>Artemisia caerulescens</i> L.: Metabolites from In Vitro Shoots and Wild Plants

Halophyte plants are potential resources to deal with the increasing soil salinity determined by climatic change. In this context, the present study aimed to investigate the germplasm conservation of Artemisia caerulescens collected in the San Rossore Estate (Pisa, Italy) through in vitro culture, biochemical properties, and the phytochemical composition of the volatile fraction of both in vitro shoots and different organs of wild plants (leaves, young and ripe inflorescences). The best medium tested for the shoot proliferation was MS, with the addition of 1 μM BA. Total polyphenol content and antioxidant activity were noticeable in both the inflorescences, while leaves and in vitro shoots showed lower amounts. Concerning the phytochemical investigation, the headspaces (HSs) and the essential oils (EOs) were characterized by oxygenated monoterpenes as the main chemical class of compounds in all samples, and with α- and β-thujone as the major constituents. However, the EOs were characterized by noticeable percentages of phenylpropanoids (23.6–28.8%), with brevifolin as the unique compound, which was not detected in the spontaneous volatile emissions of the same parts of the wild plant. Good amounts of EOs were obtained from different organs of the wild plant, comprising between 0.17% and 0.41% of the young and ripe inflorescences, respectively

Volatolomics of Three South African Helichrysum Species Grown in Pot under Protected Environment

Helichrysum decorum DC, Helichrysum lepidissimum S. Moore, and Helichrysum umbraculigerum are three species traditionally used in the South African medicine. The present work deals with the investigation of the spontaneous emission and the essential oils obtained from these plants cultivated in open field under uniform conditions. Fractions of the volatile organic compounds of the three species were rich in monoterpene hydrocarbons, representing more than 70% of the total composition. Pinene isomers were the most representative compounds: &beta;-pinene in H. decorum (53.0%), and &alpha;-pinene in H. lepidissimum (67.9%) and H. umbraculigerum (54.8%). These latter two species evidenced an important amount of sesquiterpene hydrocarbons (SH) especially represented by &gamma;-curcumene (H. lepidissimum) and &alpha;- and &beta;-selinene (H. umbraculigerum). On the contrary, in the EOs, sesquiterpenes compounds prevailed, representing more than 64% of the identified fraction to reach more than 82 and 87% in H. umbraculigerum and H. lepidissimum, respectively. Although the chemical classes and their relative abundances were comparable among the three species, the individual compounds of EOs showed large differences. In fact, caryophyllene oxide (26.7%) and &gamma;-curcumene (17.4%) were the main constituents in H. decorum, and H. lepidissimum respectively, while neo-intermedeol (11.2%) and viridiflorol (10.6%) characterized H. umbraculigerum

DataSheet_1_Characterization of the endophytic bacterial community of Bituminaria bituminosa plant grown in vitro and its interaction with the plant extract.pdf

IntroductionBituminaria bituminosa is a medicinal plant recognized for its phytochemicals, such as furanocoumarins, pterocarpans, and flavonoids. Since the secondary metabolism is influenced by the plant-endophyte interactions, the endophytic bacterial community of B. bituminosa was explored and the possible interactions with the plant were described.Materials and methodsDifferent bacterial strains were isolated from different organs of in vitro plants as shoots, roots, and seeds. The bacterial strains were identified and phenotypically characterized for different traits; strains were also exposed to different concentrations of B. bituminosa plant extract showing different susceptibility, probably determined by different secondary metabolites produced by the plant in the different organs (i.e. aerial parts and roots).Results and discussionBacterial strains showed different phenotypic characteristics; the 6 detected haplotypes were dominated by a single species related to Stenotrophomonas rhizophila. Endophytes isolated from the aerial parts produced a higher indole-3-acetic acid (IAA) amount than those of the roots, while all strains were unable to produce biosurfactants and antagonistic activity toward the other strains. The research opens new perspectives for future analysis addressed to test the susceptibility of the endophytic bacterial community of B. bituminosa toward the pure compounds extracted from the plants, and to investigate the role of these compounds on the distribution of endophytes within the different plant tissues.</p