Honeybees use propolis as a natural pesticide against their major ectoparasite.

Honeybees use propolis collected from plants for coating the inner walls of their nest. This substance is also used as a natural antibiotic against microbial pathogens, similarly to many other animals exploiting natural products for self-medication. We carried out chemical analyses and laboratory bioassays to test if honeybees use propolis for social medication against their major ectoparasite: Varroa destructor. We found that propolis is applied to brood cells where it can affect the reproducing parasites, with a positive effect on honeybees and a potential impact on Varroa population. We conclude that propolis can be regarded as a natural pesticide used by the honeybee to limit a dangerous parasite. These findings significantly enlarge our understanding of behavioural immunity in animals and may have important implications for the management of the most important threat to honeybees worldwide.

The effects of raw propolis on Varroa-infested honey bee (Apis mellifera) workers.

Self-medication plays a major role in the behavioral defense against pathogens and parasites that animals have developed during evolution. The conditions defining this adaptive behavior are: (1) contact with the substance in question must be deliberate; (2) the substance must be detrimental to one or more parasites; (3) the detrimental effect on parasites must lead to increased host fitness. Recent studies have shown that A. mellifera colonies are able to increase resin foraging rates when infested by V. destructor, whereas further investigations are needed for evidence of parasite and host fitness. In order to understand whether Varroa-infested colonies could benefit from increasing levels of resin, we carried out laboratory bioassays to investigate the effects of propolis on the fitness of infested workers. The longevity and energetic stress of adult bees kept in experimental cages and artificially infested with the mite were thus monitored over time. At the same time, in vitro experiments were performed to study the contact effects of crude propolis on Varroa mites. Our results clearly demonstrate the positive effects of raw propolis on the lifespan of Varroa-infested adult bees. A low narcoleptic effect (19-22%) of raw propolis on phoretic mites after 5 h was also observed. In terms of energetic stress, we found no differences between Varroa-free and Varroa-infested bees in terms of the daily sucrose solution demand. Our findings seem to confirm the hypothesis that resin collection and propolis use in the hive represent an example of self-medication behavior in social insects.

Effect of maturation and cold storage on the organic acid composition of myrtle fruits.

BACKGROUND: The effect of maturation and senescence on the chemical composition of two myrtle cultivars was studied in mature, overripe and cold-stored fruits in order to find the most appropriate harvesting period and best storage technology for industrial purposes. RESULTS: After cold storage at 10 °C for 15 days, berry weight loss ranged from 12.5 to 18.4%, with the highest losses in less mature fruits. Titratable acidity decreased during maturation and cold storage in both cultivars. Reducing and total sugars increased during maturation. Anthocyanin concentration increased during maturation but decreased in overripe berries. The major organic acids in myrtle fruits were quinic, malic and gluconic acids. In fresh and cold-stored fruits, malic acid rose to 3 g kg(-1) and decreased thereafter. Quinic acid peaked at 90 or 120 days after bloom and decreased thereafter to reach low concentrations in mature fruits. CONCLUSION: Cold storage for 15 days at 10 °C does not affect myrtle fruit quality for liqueur production. Anthocyanin concentration is the best indicator of harvest time for industrial purposes. Gluconic acid concentration is high in mature, overripe and cold-stored berries. This parameter can be used as a marker of the onset of fruit senescence.

Three years monitoring survey of pesticide residues in Sardinia wines following integrated pest management strategies.

This paper reports the results of a pesticide monitoring survey on wine grapes from the 2008-2010 vintage from vineyards grown according to integrated pest management strategies. A multi-residue gas chromatography-mass spectrometry method in electron ionization and chemical ionization mode has been used for the determination of 30 pesticides in wine samples. The analytical method showed good recoveries and allowed a good separation of the selected pesticides. Repeatability and intermediate precision showed good results with CV < 20 %. The instrumental method limits of determination (LOD) and of quantification (LOQ) were below the maximum residue levels set in wine. The analysis of the wines showed that pesticide residues were below the instrumental LOQ, and most of them were undetectable (

Determination of Pesticide Residues in IV Range Artichoke (Cynara cardunculus L.) and Its Industrial Wastes.

Fourth-range products are those types of fresh fruit and vegetables that are ready for raw consumption or after cooking, and belong to organic or integrated cultivations. These products are subject to mild post-harvesting processing procedures (selection, sorting, husking, cutting, and washing), and are afterwards packaged in packets or closed food plates, with an average shelf life of 5-10 days. Artichokes are stripped of the leaves, stems and outer bracts, and the remaining heads are washed with acidifying solutions. The A LC-MS/MS analytical method was developed and validated following SANTE guidelines for the detection of 220 pesticides. This work evaluated the distribution of pesticide residues among the fraction of artichokes obtained during the industrial processing, and the residues of their wastes left on the field were also investigated. The results showed quantifiable residues of one herbicide (pendimethalin) and four fungicides (azoxystrobin, propyzamide, tebuconazole, and pyraclostrobin). Pendimethalin was found in all samples, with the higher values in leaves 0.046 ± 8.2 mg/kg and in field waste 0.30 ± 6.7 mg/kg. Azoxystrobin was the most concentrated in the outer bracts (0.18 ± 2.9 mg/kg). The outer bracts showed the highest number of residues. The industrial waste showed a significant decrease in the number of residues and their concentration.

Determination of wine aroma compounds by dehydration followed by GC/MS.

A new extraction method for the analysis of the volatile fraction of white and red wines has been developed and validated. A dehydration step with MgSO4 separated an aroma compounds-rich alcoholic-glycerine layer. Spiked samples showed good recoveries in the range between 75 and 120% with CVmax% of 17, except for 2-phenylethanol and y-butyrolactone for which recoveries in red wines were under 60%, and for monoethylsuccinate, where recoveries averaged 50 and 60% in white and red wines, respectively. Method repeatability and intermediate precision showed good CVmax% with minimum and maximum values between 7.7 and 24, and between 18.7 and 25.0, respectively. The average determination coefficients were greater than 0.99 with CVmax% of 13. The instrumental LOD and LOQ were, in all cases, under 0.05 mg/L, except for 2,3 butanediol (0.20 mg/L). Overall, the presence of wine matrix affected aroma compounds responses in GC/MS analysis. These observations indicate that the use of a matrix-matched calibration curve is mandatory to obtain reliable quantitative data.

Influence of Salting Technology on the Diffusion of NaCl in Swordfish (Xiphias gladius) Fillets.

Swordfish is the most widespread billfish in the aquatic environment. The industrial processing of swordfish fillets involves salting, drying, and smoking steps. Salting techniques, dry or wet, are the most common method of fish preservation. This work evaluated salt diffusion in swordfish fillets after traditional dry salting and wet industrial injection salting methods. The data obtained from the dry salting studies highlighted that the salt diffusion process in swordfish meat was an unfavorable process depending on the contact time with the salt/meat. Moreover, irregularly shaped fillets negatively affected the salt migration in the different areas, leading to inhomogeneous and possibly unsafe final products. On the contrary, wet injection salting was suitable for processing swordfish fillets. As a result, the final products had a homogeneous salt concentration, maintained the organoleptic characteristics and health benefits for a long period, and achieved a longer shelf-life. Furthermore, the water activity (aw) values detected for the different processed fillets confirmed the physicochemical features of the final products and allow the classification of safe products. Moreover, injection salting is a quick process compatible with industrial production times.

Effect of High-Pressure Processing on Fresh Sea Urchin Gonads in Terms of Shelf Life, Chemical Composition, and Microbiological Properties.

Paracentrotus lividus is a widespread sea urchin species appreciated worldwide for the taste of its fresh gonads. High-pressure processing (HPP) can provide a thermal equivalent to pasteurization, maintaining the organoleptic properties of the raw gonads. This study evaluated HPP technology's effect at 350 MPa and 500 MPa on microbial inactivation and biochemical characteristics of P. lividus gonads. HPP at 350 MPa resulted in a higher decrease in protein and free amino acids associated with a loss of olfactory, color, and gustatory traits and a visual alteration of the texture. On the other hand, gonad samples stored for 40 days after treatments at 500 MPa showed a good organoleptic profile similar to fresh gonads. Furthermore, only 500 MPa effectively reduced mesophilic bacteria contamination among the two HPP treatments carried out. Total lipids increased during storage; however, the SAFA/PUFA rate was homogeneous during HPP trials ranging from 2.61-3.91 g/100 g. Total protein decreased more than 40% after HPP at 350 MPa, whereas, after 500 MPa, it remained stable for 20 days. The amount of free amino acid constantly decreased during storage after HPP at 350 MPa and remained constant at 500 MPa. HPP can effectively remove the bacterial flora and inactivate enzymes, maintaining the properties of the fresh sea urchin gonads.

Influence of Different Light Sources on the Biochemical Composition of Arthrospira spp. Grown in Model Systems.

Arthrospira platensis and Arthrospira maxima are prokaryotic microalgae commercially marketed as spirulina. The pigments extracted from these algae are widely used for cosmetic and nutraceutical applications. This work aimed to evaluate the influence of three light-emitting lamps (white, orange and blue) on the growth and biomass composition of two strains of A. platensis (M2 and M2M) and one of A. maxima. The obtained data show strain- and light-dependent responses of the microalgae. In addition, white and orange lights led to a similar overall effect by increasing the levels of chlorophyll a and carotenoids. However, exposure to orange light resulted in the highest dry weight (5973.3 mg L-1 in M2M), whereas white light stimulated an increase in the carbohydrate fraction (up to 42.36 g 100 g-1 in A. maxima). Conversely, blue light led to a constant increase in the concentration of phycocyanin (14 g 100 g-1 in A. maxima) and a higher content of proteins in all strains. These results provide important environmental information for modulating the growth of different spirulina strains, which can be used to address the synthesis of biochemical compounds of strategic importance for the development of new nutraceutical foods.

Influence of fenamidone, indoxacarb, pyraclostrobin, and deltamethrin on the population of natural yeast microflora during winemaking of two sardinian grape cultivars.

The influence of fenamidone ((S)-1-anilino-4-methyl-2-methylthio-4-phenylimidazolin-5-one), pyraclostrobin (methyl 2-[1-(4-chlorophenyl)pyrazol-3-yloxymethyl]-N-methoxycarbanilate), indoxacarb (methyl 7-Chloro-2,5-dihydro-2-[[(methoxycarbonyl) [4- (trifluoromethoxy) phenyl] amino] carbonyl] indeno[1,2-e][1,3,4] oxadiazine-4a(3H)-carboxylate), and deltamethrin ([cyano-[3-(phenoxy)phenyl]methyl] 3-(2,2-dibromoethenyl)-2,2-dimethylcyclopropane-1-carboxylate) on spontaneous fermentation carried out by natural yeast grapes microflora, was studied during the wine-making process. Aliquots of pesticide standard solutions were added to the grapes before crushing, to reach a concentration equal or half the maximum residue limit (MRL). Vinifications were performed, with maceration (R), or without maceration (W). During the wine-making process, samples were taken at the beginning (one hour after grapes crushing), at the middle and at the end of the spontaneous fermentation process. At half the MRL concentration, deltamethrin affected Pichia sp. population with a decrease of almost 50 %, while fenamidone decreased Candida sp., Candida stellata at 83, and 36%, respectively. Metschnikowia pulcherrima population decreased in all samples when compared to the control. Experiments at MRL levels showed a strong reduction for all non-Saccharomyces yeast species, when grapes had been treated with pyraclostrobin, fenamidone, and deltamethrine, except for Candida sp. which was found to have been affected only by fenamidone residues. Growth zone inhibition test showed only an in vitro activity of pyraclostrobin over Kloeckera spp., C. stellata, and M. pulcherrima. Microvinification experiments produced wines with no differences concerning S. cerevisiae population as well as production of ethanol and residual sugars. Experiments showed that at the end of the fermentation process pesticides were adsorbed by the lees and grape skins, and no pesticides residue was detectable in wine.

Chlorpyrifos residues levels in fruits and vegetables after field treatment.

Chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate) was applied with three different formulations on oranges, peaches, tomatoes, wine and table grapes, and its behaviour was evaluated after field treatment. The formulations applied were emulsifiable concentrates (EC), microencapsulates (ME), and wettable granules (WG). The residues were similar in all crops studied in the EC and WG experiments, except peaches with WG treatment, the residue amount was lower than EC values. Tomatoes which were grown in greenhouse showed similar residues in all treatments just after treatment. Wine and table grapes showed different decline curves in the EC experiments ascribable to the different growing technology. Instrumental limit of determination (LOD) and limit of quantification (LOQ) for all matrices were 0.01, and 0.03 mg kg⁻¹, respectively. Repeated treatments showed that Chlorpyrifos can accumulate leading to residue levels at the preharvest interval (PHI) over the maximum residue level (MRL), especially on oranges and peaches. Among the formulates used ME showed the higher risk of residues over the MRL at harvest.

Effect of the Technological Process from Vine to Wine on Pesticide Residues in Vernaccia di Oristano Cultivar.

Vernaccia is a white grape mostly used to produce a distinct wine protected by the controlled designation of origin (DOC) recognition. It is very susceptible to fungal disease, and it is subjected to a defined management protocol in the field. Winemaking could influence pesticide residues through different mechanisms. This work investigated the influence on pesticide residues of the winemaking process at the industrial level of the wine Vernaccia di Oristano. Thirty-five samples of grapes, two musts, and two vines (both liquid fraction and pellets) were analyzed by using a validated multiresidue LC-MS/MS (Liquid Chromatography with tandem mass spectrometry) method. Data obtained showed the presence in grapes juice of 16 pesticides (8 not allowed in the EU) with mandipropamid and mepanipyrim, the only ones with values higher than their MRL (maximum residue level). Pesticide residues decrease in must was related to the dilution effect due to mixing the grape samples. However, pellets analysis also confirmed the high affinity of pesticides for the suspended material (fenhexamid), whereas the increase in wine to a re-solubilization process from the lees during the fermentation step. The present paper highlighted the effectiveness of the technological process of winemaking to decrease pesticide residues compared to the raw material.

In Vitro Activity of Several Essential Oils Extracted from Aromatic Plants against Ascosphaera apis.

The use of natural substances such as essentials oils against bee pathogens is of great interest as an alternative to traditional methods based on synthetic compounds like antibiotics and fungicides, in order to minimize the risk of having toxic residues in hive products and to prevent the development of resistance phenomena. This study evaluated the inhibitory, fungicidal and sporicidal activity of ten essential oils extracted from aromatic plants against Ascosphaera apis, the etiological agent of chalkbrood, an invasive honey bee mycosis. The most effective essential oils were Thymus herba-barona, Thymus capitatus and Cinnamomum zeylanicum, which showed values of minimum fungicidal concentration and minimum sporicidal concentration ranging from 200 to 400 ppm. Carvacrol was the main component of Thymus capitatus and Thymus herba-barona oils, whereas cinnamic aldehyde prevailed in Cinnamomum zeylanicum oil. Further in-apiary studies will allow the evaluation of side effects on bees and residues in hive products.

Flash flood simulation and valve behavior of Mytilus galloprovincialis measured with Hall sensors.

Mussels close their shell as a protective strategy and the quantification of this behavioral marker may represent an alarm signal when they are exposed to environmental stressors. In the present study, we investigated the ability of the Mediterranean mussel Mytilus galloprovincialis to recover and then the resilience or inertia of valve activity after a pulsing exposition to diverse levels of salinity (5, 10, 20, and 35 PSU as reference value). The trial simulated an event of drastic and sudden reduction of seawater salinity thus mimicking an event of flash flood from intense rain. Valve gaping and movements were measured in continuous cycle for 10 days using a customized magneto-electric device which uses Hall sensors. Results showed that under normal conditions of salinity (35 PSU), the general pattern of valve movements was a continuously open state with sporadic spikes indicating a closing motion. At salinity of 5, PSU mussels reacted by closing their valves, leading to a 77% mortality on the 4th day. At salinity of 10, PSU animals were observed with closed valves for the entire duration of the exposure and no mortality occurred, they showed a significant reduction in the valve activity once the reference value of salinity was re-established. In contrast, salinity of 20 PSU did not trigger a significant behavioral response. Interestingly, there no define rhythms of valve movements were recorded during salinity challenges.

Fipronil and Fipronil Sulfone Distribution in Chicken Feathers and Eggs after Oral and Dermal Exposure.

This work aimed to investigate the bio-distribution and the persistence of fipronil and its primary metabolite fipronil sulfone after oral and dermal administration by simulating natural farming conditions. Fipronil and fipronil sulfone detection and quantification were performed in different poultry matrices using an LC-MS/MS method coupled with modified QuEChERS extraction. After oral administration, fipronil was detected in feathers at each sampling time, in eggs for 28 days, and in the internal organs at the end of the experiment. After dermal administration, high levels of fipronil were detected in feathers, accounting for 195.85 ± 8.54 mg/kg, which were reduced by a third after 60 days. No traces of fipronil were detected in the eggs or internal organs. In addition, fipronil sulfone showed remarkable residues in all samples in trial 2. The data obtained confirmed that inappropriate use of unauthorized pesticides can lead to severe contamination of entire poultry farms. The contemporary presence of fipronil sulfone in feathers and eggs associated with the lack of fipronil in eggs suggests recent dermal contamination or past oral contamination. Moreover, simultaneous analysis of hens' feathers and eggs could represent a new method to improve large-scale monitoring programs and animal welfare, limiting their slaughter.

Zoxamide accumulation and retention evaluation after nanosuspension technology application in tomato plant.

BACKGROUND: Low water solubility of pesticide requires formulations with high levels of stabilizers and organic solvents. Moreover, only 0.1% of the applied pesticides formulation reaches the target, while 99.9% spreads in the surrounding environment. Therefore, there is the need for more efficient and environmentally sustainable alternatives. RESULTS: Zoxamide (ZO) nanosuspension was prepared through a media milling technique by using the stabilizer polysorbate 80. The thin and acicular crystals obtained, showed particle size of 227 nm, polydispersion index of 0.247 and zeta potential of -28 mV. Dimensional data and morphology of ZO nanocrystals alone, on tomato leaves and berries, were confirmed by scanning electron microscopy. The reduction in size for ZO crystals obtained after the milling process increased pesticide water solubility till 39.6 mg L-1 , about 1.6 the solubility obtained with a conventional commercial formulation. Field and dip contamination trials performed on tomato plants showed the nanosuspension's ability to increase ZO deposition and accumulation versus a coarse ZO suspension and commercial formulation, respectively. CONCLUSIONS: The nanoformulation proposed, resulted in low cost and was easy to make. Moreover, the organic solvent-free composition together with a low surfactant addition assured a minor environmental impact. Finally, the increased retention and deposition of the fungicide can reduce the amounts of ZO formulation applied to tomatoes. © 2021 Society of Chemical Industry.

Integrated environmental evaluation of heavy metals and metalloids bioaccumulation in invertebrates and seaweeds from different marine coastal areas of sardinia, mediterranean sea.

In this work, three gastropods Patella vulgata, Osilinus turbinata, and Tahis clavigera, one echinoderm Parancetrotus lividus, one coelenterate Anemonia sulcata, and two seaweed Padina pavonica, and Cystoseira mediterranea were collected from three different marine areas of Sardinia in the Mediterranean sea and studied for heavy metals and metalloid content and accumulation trends. Inductively coupled plasma optical emission spectrometry (ICP-OES) was used for the determination of Al, AS, B, Ba, Be, Cd, Co, Cr, Cu, Fe, Hg, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Te, Ti, V and Zn in the selected samples. The results showed that gastropods were capable of accumulating Al, Ba, Cu, Fe, Sr, and Zn; seaweeds can better concentrate Al, Fe, and Zn than all other species. At the same time, echinoderms and coelenterate had limited ability to store specific metals, showing a much more homogeneous distribution. PCA analysis allowed us to discriminate among the sites and the species. Cala Zafferano was the area with the higher values of accumulation of all metals in all species as expected, considering its proximity to industrial sites. The results of the analysis showed clearly that heavy metal and metalloid accumulation was different for each species studied. Therefore, for a correct environmental assessment of a given area, a comprehensive approach is strongly recommended by exploiting the different properties of both accumulation and concentration of the metals by different aquatic species.

Do Best-Selected Strains Perform Table Olive Fermentation Better than Undefined Biodiverse Starters? A Comparative Study.

Twenty-seven Lactobacillus pentosus strains, and the undefined starter for table olives from which they were isolated, were characterised for their technological properties: tolerance to low temperature, high salt concentration, alkaline pH, and olive leaf extract; acidifying ability; oleuropein degradation; hydrogen peroxide and lactic acid production. Two strains with appropriate technological properties were selected. Then, table olive fermentation in vats, with the original starter, the selected strains, and without starter (spontaneous fermentation) were compared. Starters affected some texture profile parameters. The undefined culture resulted in the most effective Enterobacteriaceae reduction, acidification and olive debittering, while the selected strains batch showed the lowest antioxidant activity. Our results show that the best candidate strains cannot guarantee better fermentation performance than the undefined biodiverse mix from which they originate.

Propolis Consumption Reduces Nosema ceranae Infection of European Honey Bees (Apis mellifera).

Nosema ceranae is a widespread obligate intracellular parasite of the ventriculus of many species of honey bee (Apis), including the Western honey bee Apis mellifera, in which it may lead to colony death. It can be controlled in A. mellifera by feeding the antibiotic fumagillin to a colony, though this product is toxic to humans and its use has now been banned in many countries, so in beekeeping, there exists a need for alternative and safe products effective against N. ceranae. Honeybees produce propolis from resinous substances collected from plants and use it to protect their nest from parasites and pathogens; propolis is thought to decrease the microbial load of the hive. We hypothesized that propolis might also reduce N. ceranae infection of individual bees and that they might consume propolis as a form of self-medication. To test these hypotheses, we evaluated the effects of an ethanolic extract of propolis administered orally on the longevity and spore load of experimentally N. ceranae-infected worker bees and also tested whether infected bees were more attracted to, and consumed a greater proportion of, a diet containing propolis in comparison to uninfected bees. Propolis extracts and ethanol (solvent control) increased the lifespan of N. ceranae-infected bees, but only propolis extract significantly reduced spore load. Our propolis extract primarily contained derivatives of caffeic acid, ferulic acid, ellagic acid and quercetin. Choice, scan sampling and food consumption tests did not reveal any preference of N. ceranae-infected bees for commercial candy containing propolis. Our research supports the hypothesis that propolis represents an effective and safe product to control N. ceranae but worker bees seem not to use it to self-medicate when infected with this pathogen.

Influence of the Technological Process on the Biochemical Composition of Fresh Roe and Bottarga from Liza ramada and Mugil cephalus.

Bottarga is a high-priced delicacy with high nutritional value, and, in Italy, bottarga from mullets has been recognized to be a traditional food product. The flathead grey mullet Mugil cephalus and the thinlip grey mullet Liza ramada are the main cultured grey mullets in the Mediterranean Sea. In this study, fresh roe and bottarga from these two species were investigated to evaluate the influence of the technological process and the species on their biochemical composition and health advantages. The 1 h/200 g salting-out step did not increase the levels of NaCl in the bottarga, although it highly decreased the levels of some heavy metals like Cu and Al. Processing of fresh roe in bottarga led to an essential modification of the lipid fraction, following a general series of monousatturated fatty acid (MUFA)> poliunsutturated fatti acid (PUFA) > saturated fatty acid (SAFA) and an increase in both ω3 and ω6 in Liza ramada. Moreover, bottarga showed higher levels of squalene and cholesterol and an increased Essential Amino Acid/Total Amino Acid ratio (EAA/TAA) in both species. In addition to the nutritional benefits for the consumer, the process proposed in this study may represent a reliable tool for local producers to obtain a final bottarga with both a reproducible biochemical composition and organoleptic characteristics.