Fingerprinting Chemical Markers in the Mediterranean Orange Blossom Honey: UHPLC-HRMS Metabolomics Study Integrating Melissopalynological Analysis, GC-MS and HPLC-PDA-ESI/MS.

(1) Background: Citrus honey constitutes a unique monofloral honey characterized by a distinctive aroma and unique taste. The non-targeted chemical analysis can provide pivotal information on chemical markers that differentiate honey based on its geographical and botanical origin. (2) Methods: Within the PRIMA project "PLANT-B", a metabolomics workflow was established to unveil potential chemical markers of orange blossom honey produced in case study areas of Egypt, Italy, and Greece. In some of these areas, aromatic medicinal plants were cultivated to enhance biodiversity and attract pollinators. The non-targeted chemical analysis and metabolomics were conducted using ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS). (3) Results: Forty compounds were disclosed as potential chemical markers, enabling the differentiation of the three orange blossom honeys according to geographical origin. Italian honey showed a preponderance of flavonoids, while in Greek honey, terpenoids and iridoids were more abundant than flavonoids, except for hesperidin. In Egyptian honey, suberic acid and a fatty acid ester derivative emerged as chemical markers. New, for honey, furan derivatives were identified using GC-MS in Greek samples. (4) Conclusions: The application of UHPLC-HRMS metabolomics combined with an elaborate melissopalynological analysis managed to unveil several potential markers of Mediterranean citrus honey potentially associated with citrus crop varieties and the local indigenous flora.

Precision Beekeeping Systems: State of the Art, Pros and Cons, and Their Application as Tools for Advancing the Beekeeping Sector.

The present review aims to summarize the more recent scientific literature and updated state of the art on the research effort spent in adapting hardware-software tools to understand the true needs of honeybee colonies as a prerequisite for any sustainable management practice. A SWOT (Strengths, Weaknesses, Opportunities, and Threats) analysis was also performed with the aim of identifying the key factors that could support or impair the diffusion of precision beekeeping (PB) systems. Honeybee husbandry, or beekeeping, is starting to approach precision livestock farming (PLF), as has already happened in other animal husbandry sectors. A transition from the current paradigm of rational beekeeping to that of precision beekeeping (PB) is thus expected. However, due to the peculiarities of this species and the related farming practices, the PB technological systems (PB systems) are still undergoing a development process that, to some extent, limits their large-scale practical application. Several physical-chemical (weight, temperature, humidity, sound, gases) and behavioral traits (flight activity, swarming) of the hive are reviewed in light of the evolution of sensors, communication systems, and data management approaches. These advanced sensors are equipped with a microprocessor that records data and sends it to a remote server for processing. In this way, through a Wireless Sensor Network (WSN) system, the beekeeper, using specific applications on a personal computer, tablet, or smartphone, can have all the above-mentioned parameters under remote control. In general, weight, temperature, and humidity are the main hive traits monitored by commercial sensors. Surprisingly, flight activity sensors are rarely available as an option in modular PB systems marketed via the web. The SWOT analysis highlights that PB systems have promising strength points and represent great opportunities for the development of beekeeping; however, they have some weaknesses, represented especially by the high purchasing costs and the low preparedness of the addressed operators, and imply some possible threats for beekeeping in terms of unrealistic perception of the apiary status if they applied to some hives only and a possible adverse impact on the honeybees' colony itself. Even if more research is expected to take place in the next few years, indubitably, the success of commercial PB systems will be measured in terms of return on investment, conditioned especially by the benefits (higher yields, better colonies' health) that the beekeeper will appraise as a consequence of their use.

A Multipurpose Leguminous Plant for the Mediterranean Countries: Leucaena leucocephala as an Alternative Protein Source: A Review.

In tropical and subtropical regions, as well as in the internal and/or marginal Mediterranean areas, one of the most important problems related to animal production is represented by the inadequate nutritional supplies. The low productivity of the animals, often connected to reduced annual growth, is, in fact, not infrequently attributable to the low nitrogen content and the high fiber content of the local plant species and crop residues that constitute the base ingredients of the rations commonly adopted by farmers. The use of the supplementation with arboreal and shrub fodder, although often containing anti-nutritional factors and toxins that limit its use, could be a profitable way to alleviate the nutritional deficiencies of the basic diets. Leucaena leucocephala (Lam.) De Wit is native to Central America and widely naturalized in the majority of Latin American countries. It is a legume suitable for tropical and subtropical environments including the countries of the Mediterranean area. Moreover, its spread is desirable if we consider the multiple uses to which it is suitable, the considerable amount of biomass produced, and its role in preserving the environment. The aim of this work was to highlight the characteristics of Leucaena that can justify its wide diffusion. A structured analysis of strengths and weaknesses was performed accordingly. Being a good protein source for feeding livestock, it could be a species to be introduced in the inland areas of the Mediterranean countries as an alternative protein source; the limit represented by the presence of anti-nutritional factors could be overcome by feed processing and by launching targeted research programs.

Carbon Footprint of Mediterranean Pasture-Based Native Beef: Effects of Agronomic Practices and Pasture Management under Different Climate Change Scenarios.

A better understanding of soil organic carbon (SOC) dynamics is needed when assessing the carbon footprint (CFP) of livestock products and the effectiveness of possible agriculture mitigation strategies. This study aimed (i) to perform a cradle-to-gate CFP of pasture-based beef cattle in a Mediterranean agropastoral system (ii) and to assess the effects on the CFP of alternative tillage, fertilizing, and grazing practices under current (NCC) and future climate change (CC) scenarios. Minimum (Mt) and no-tillage (Nt) practices were compared to current tillage (Ct); a 50% increase (Hf) and decrease (Lf) in fertilization was evaluated against the current (Cf) rate; and rotational grazing (Rg) was evaluated versus the current continuous grazing (Cg) system. The denitrification-decomposition (DNDC) model was run using NCC as well as representative concentration pathways to investigate the effects of farm management practices coupled with future CC scenarios on SOC dynamics, N2O fluxes, and crop yield. Within NCC and CtCf, an emission intensity of 26.9 ± 0.7 kg CO2eq per kg live body weight was estimated. Compared to Ct, the adoption of Mt and Nt reduced the CFP by 20% and 35%, respectively, while NtHf reduced it by 40%. Conservation tillage practices were thus shown to be effective in mitigating greenhouse gas emissions.

Sowing Date and Seeding Rate Affect Bioactive Compound Contents of Chickpea Grains.

Chickpea grains may represent an alternative to soybean meals and energetic concentrates in animal feeding, as their nutritional value can help to increase the sustainability of livestock systems. Unfortunately, the presence of bioactive compounds with anti-nutritional effects can prevent its direct use, especially in mono-gastrics. It is known that the synthesis of these compounds depends on genetic expression, which is also influenced by growth conditions. The objective of this two-year study was to assess the effect of sowing date (winter versus spring) and seeding rate (70 versus 110 seeds m-2) on the accumulation of soluble carbohydrates, α-galactosides, trypsin inhibitors, and inositol phosphates in the grains of two Kabuli cultivars, in the Mediterranean climate. The results showed that seeds collected from winter sowing contained more trypsin inhibitors than those seeded in spring (+ 4%, on average), reaching values between 16.1 and 18.6 TIU mg protein-1. The seeding rate affects only the α-galactosides content, which increases (+9%) at lower densities (70 seeds m-2). These findings suggest that agronomic management can be used to modulate the content of some anti-nutritional factors in the seeds, even though the genetic characteristics and phenotypic expression, in relation to the climatic conditions, seem to deeply affect the content of all the bioactive compounds investigated.

The Effects of Diet Formulation on the Yield, Proximate Composition, and Fatty Acid Profile of the Black Soldier Fly (Hermetia illucens L.) Prepupae Intended for Animal Feed.

The black soldier fly (BSF; Hermetia illucens L.) is a very promising insect species due to the ability to convert low-value substrates in highly nutrient feed. This work aimed to study the effect of three nominally isoenergetic diets containing plant ingredients such as barley, alfalfa, and wheat byproducts, formulated to be higher in nonfiber carbohydrates (TMD1), fibers (TMD2), and protein (TMD3) in comparison to an extensively genetic modified cereal (cornmeal)-based diet (C), on the growth, yield, and nutritive traits of BSF prepupae (BSFPs). Three growing trials with four biological replicates were carried out. Proximate and fatty acid analyses were performed on the diets and BSFPs. Feed conversion ratios (FCR), dry matter and nutrient yields, and apparent concentration factors (aBCF) for fatty acids (FAs) were calculated. Diet formulation had a substantial effect on the survival, development rate, and larval yield, but the FCR was unaffected. The BSFPs fed TMD3 did not result in a higher crude protein content in comparison to the C or TMD2 diets. Despite the leveled fat content of the diets, BSFPs reared on TMD1 were highest in fat, saturated FA, and fat yield. An apparent bioconcentration factor (aBCF) value lower than unity that was found for the unsaturated FA suggests that the BSFPs inefficiently absorb them from the diet or possibly turn them into saturated FA. However, the unsaturated FA accumulation in BSFPs depended on the levels that were found in the diet, which suggested some possibilities for the FA profile modulation. Overall, the TMD2 performed well despite the low-value of its main ingredients and high fiber content and can be considered to be a feasible option for the mass rearing of BSFPs that are intended for animal feed.

Stable isotopes in tissues discriminate the diet of free-living wild boar from different areas of central Italy.

The use of isotopic signatures in animal tissues provides information on the environment where they are living and, notably, on their diet. Carbon and, whenever possible, nitrogen stable isotope analyses were performed in animal hairs, muscles and fat. Particularly, we analyzed both carbon and nitrogen isotopic compositions (δ13C and δ15N) on wild boar samples across three different areas of central Italy (Latium region): Tyrrhenian Coast (TC), Maremma (MA) and Central Plains (CP). The agricultural habits of these areas imply that, in winter, no crops are available for wild boars, which feed mainly on acorns and natural feeds (tubers, earthworms etc.). In addition, the three areas were influenced by oak masting. One of these areas (CP) was characterised by the spreading of corn during the hunting season to attract the animals. For each area, we sampled 10 animals aged between 12 and 24 months and balanced by gender. Anenrichment of δ13C in CP area, where corn was used, was observed in all the analysed tissues in comparison to other areas (MA and TC). In CP area, enriched values of δ15N were also observed in all the tissues. The research demonstrates that both δ13C andδ15N in free-living wild boar tissues are influenced by sampling area. According to feeding habits of the species and wildlife management (feed supplementation), the differences observed in δ13C and δ15Nare based on the specific feeding regime; particularly the use of corn in wintertime. Furthermore, the research highlights and discusses diversities and relationships among δ13C and δ15N in the hair, fat and muscles of free-living wild boar.

Analysis of Quality-Related Parameters in Mature Kernels of Polygalacturonase Inhibiting Protein (PGIP) Transgenic Bread Wheat Infected with Fusarium graminearum.

Fusarium head blight, caused by the fungus Fusarium graminearum, has a detrimental effect on both productivity and qualitative properties of wheat. To evaluate its impact on wheat flour, we compared its effect on quality-related parameters between a transgenic bread wheat line expressing a bean polygalacturonase inhibiting protein (PGIP) and its control line. We have compared metabolic proteins, the amounts of gluten proteins and their relative ratios, starch content, yield, extent of pathogen contamination, and deoxynivalenol (DON) accumulation. These comparisons showed that Fusarium significantly decreases the amount of starch in infected control plants, but not in infected PGIP plants. The flour of PGIP plants contained also a lower amount of pathogen biomass and DON accumulation. Conversely, both gluten and metabolic proteins were not significantly influenced either by the transgene or by fungal infection. These results indicate that the transgenic PGIP expression reduces the level of infection, without changing significantly the wheat seed proteome and other quality-related parameters.

Aflatoxin B1 and fumonisin B1 affect the oxidative status of bovine peripheral blood mononuclear cells.

Mycotoxins are secondary metabolites having a high cytotoxic potential. They are produced by molds and released in food and feed. To date, the mechanisms underlying the mycotoxin-induced cytotoxicity have not been fully clarified. The induction of oxidative stress, as a possible mechanism, has been postulated. This in vitro study was focused on the effect of two widely occurring mycotoxins, aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)), on the oxidative status of bovine peripheral blood mononuclear cells (PBMC) incubated for 2 and 7 days at different levels of AFB(1) (0, 5 and 20 µg/ml) and FB(1) (0, 35 and 70 µg/ml). Reactive oxygen metabolites (ROM), intracellular thiols (SH), malondialdehyde (MDA) and gene expression of cytoplasmic superoxide dismutase (SOD) and glutathione peroxidase (GSHPX-1) were measured on PBMC after incubation. The highest concentration of AFB(1) and all concentrations of FB(1) caused an increase (p<0.05) of intracellular ROM without any time dependent effect. Intracellular SH decreased with 20 µg AFB(1)/ml (p<0.05) and the effect was particularly marked after 7 days of exposure. Intracellular SH were not affected by FB(1) even though a lower (p<0.05) SH level after 2 days exposure than after 7 days was observed. MDA increased (p<0.05) in AFB(1) or FB(1) treated PBMC. The exposure to FB(1) for 7 days increased MDA (p<0.05) only in cells treated with 70 µg/ml. Exposure of PBMC to AFB(1) reduced SOD mRNA while FB(1) decreased both SOD and GSHPX-1 mRNA abundance. These results demonstrate that, even though by different mechanisms, AFB(1) and FB(1) may induce cytotoxicity through an impairment of the oxidative status of PBMC.