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Atlante per l'identificazione del polline di oltre 50 fanerofite presenti nel territorio della Riserva Naturale "Bosco della Fontana" (Marmirolo, provincia di Mantova)

In alcune aree geografiche, gli allergeni minori, in particolare i panallergeni, possono essere determinanti per l'insorgenzae/o l'amplificazione dei sintomi di molti pazienti. Tra i panallergeni biochimicamente più studiati ci sono le profiline. In questo lavoro, analizziamo quanto le profiline siano effettivamente un importante componente atmosferico e se la loro presenza è correlata alla concentrazione del polline.

Scopo del presente lavoro è analizzare una serie storica ventennale di concentrazioni giornaliere del polline delle Fagaceae, registrate nell'area urbana di Sassari, e studiare le relazioni con i parametri meteorologici temperatura e umidità dell'aria e precipitazioni. L'analisi ha riguardato una serie storica di 23 anni (1986-2008). Le concentrazioni polliniche sono state rilevate a scala giornaliera da un campionatore Spore-Trap, Burkard e i dati meteorologici sono stati rilevati da una stazione ubicata in prossimità del campionatore di polline. In particolare, sono state analizzate le variazioni della data di inizio e di fine della stagione pollinica, il momento del picco di massima concentrazione e la durata della stagione pollinica. In generale, negli anni considerati, l'inizio della stagione pollinica si è verificato tra la fine di marzo (30/3/2003) e la fine di aprile (28/4/1988-1990); la data di fine stagione è stata osservata tra la fine di maggio (29/5/1986) e la seconda decade di luglio (19/7/2000); mentre il momento del picco di massima concentrazione pollinica si è verificato tra la fine di aprile (25/4/89) e l'inizio di giugno (6/6/99). Nei ventitré anni analizzati, la durata della stagione pollinica è risultata molto variabile con un valore medio pari a 69 giorni, una durata minima di 40 giorni nel 1994 e una massima di 97 nel 2002. Il valore medio della concentrazione pollinica annua è stato di 5336 (granuli/m3), il minimo di 550 (granuli/m3) nel 1986 e il valore massimo di 8678 (granuli/m3) nel 2001. Le date in cui si è verificato il picco di massima concentrazione di polline nei vari anni è risultato positivamente correlato con la temperatura dell'aria, mentre la concentrazione di polline è risultata correlata negativamente con l'umidità relativa e le precipitazioni. I risultati ottenuti in questo lavoro, confermano quanto riportato da altri autori sulle caratteristiche delle stagioni polliniche delle Fagaceae in ambiente Mediterraneo.

Laboratory, field and semi-field experiments have been carried out on the predatory mites Kampimodromus aberrans, Amblyseius andersoni, Typhlodromus pyri and Phytoseius finitimus in order to optimize biological control strategies against phytophagous mites in apple orchards. The first experiment concerned the augmentative releases of K. aberrans in organic and conventional apple orchards. The most frequent insecticides used in organic orchards were pyrethrins and spinosad, whereas neonicotinoids, OPs and IGRs were used in conventional orchards. K. aberrans releases were successful and predatory mites were significantly higher in released than in control plots. Moreover, populations were high in organic orchards compared to conventional orchards. Amblyseius andersoni was naturally occurring in all orchards. In 2010, no effect of K. aberrans release and orchard management were observed on A. andersoni populations but in 2011 A. andersoni population densities were low in released plots compared to control plots. In organic orchards located at Spresiano releases were made on two cultivars, i.e., Florina and Golden Delicious. In both 2010 and 2011 releases, K. aberrans population densities were higher in release than in control plots of both cultivars. K. aberrans population increased during the three years on Florina but not in Golden Delicious orchard. On the latter A. andersoni population densities were lower in released than in control plots. In conventional orchards K. aberrans did not establish probably because of a series of non-selective insecticide and fungicide treatments. However, the use of spinosad was associated to a decline of predatory mite populations in organic orchards. The second experiment considered interspecific predation among K. aberrans, A. andersoni, T. pyri and P. finitimus. All four predators fed with heterospecific larvae were able to survive some weeks laying a number of eggs. A. andersoni consumed more T. pyri and P. finitimus than K. aberrans larvae and its fecundity was higher on the first two prey species. A diet based on T. pyri larvae was associated to the highest conversion rate of food into egg biomass; longevity was higher for predatory mite females fed with K. aberrans larvae. K. aberrans preyed more P. finitimus than T. pyri and A. andersoni larvae and its fecundity resulted higher on the first species. The conversion of food into egg biomass did not show precise trend while the longevity of K. aberrans resulted higher when fed with A. andersoni. Typhlodromus pyri consumed more P. finitimus and K. aberrans than A. andersoni larvae and laid more eggs on the first two species. Phytoseius finitimus preyed more K. aberrans and T. pyri than A. andersoni larvae but fecundity was not affected by prey; predatory mite females fed with A. andersoni larvae were more efficient in converting food into egg biomass but those fed with K. aberrans larvae lived longer. In terms of predation rate and fecundity, A. andersoni seems to be advantaged over T. pyri, T. pyri over K. aberrans, and A. andersoni, K. aberrans and T. pyri over P. finitimus. The performance of A. andersoni, K. aberrans and T. pyri in terms of predation rate and fecundity proved to be better of P. finitimus larvae than on other prey. The low prey consumption and fecundity of the latter suggest that is disadvantaged in interspecific predation. The comparison between pollen and prey diets confirmed the positive effect of pollen on the fecundity of all four predatory mite species. The third experiment deals with the effects of insecticides, frequently applied in apple orchards, on K. aberrans in field and laboratory conditions. Insecticides including chlorpyrifos, thiacloprid, acetamiprid, lufenuron, indoxacarb, methoxyfenozide and etofenprox were tested. In field studies etofenprox was the most detrimental pesticide to predatory mites. It caused dramatic effects on K. aberrans populations and induced spider mite outbreaks. In other trials tau-fluvalinate and spinosad did similar effects. In laboratory studies we assessed lethal and sub-lethal effects for each pesticide tested. Spinosad, tau-fluvalinate, thiamethoxam, clothianidin and imidacloprid were included. Etofenprox, spinosad with tau-fluvalinate proved to be harmful insecticides to K. aberrans in terms of lethal effects. Neonicotinoids did not affect survival so much but their sub-lethal effects were more prominent in terms of fecundity. Other pesticides such as chlorpyrifos, indoxacarb, lufenuron and methoxyfenozide also significantly reduced fecundity in K. aberrans at a lower level. In the fourth study we explored the role of pollen in alleviating the effects of pesticides on K. aberrans at individual and population levels. In the first part we tested biological and demographic parameters of K. aberrans on pollen and Panonychus ulmi. K. aberrans was reared individually on P. ulmi and on pollen. Both food sources allowed for predatory mites survival, development and reproduction. Pollen was preferred food as compared to P. ulmi. Total fecundity and survival rates of K. aberrans were higher on pollen than on tetranychids. Developmental times of protonymphs, deutonymphs and adults of K. aberrans were longer on P. ulmi than on pollen. Life-table parameters of K. aberrans were positively affected by pollen. The second part deals with the interactions of pesticides, pollen and K. aberrans in semi-field conditions. Experimental factors were: insecticide application and pollen application . Predatory mites were released on 3-4 shoots per plant 7 days before insecticide applications. Typha latifolia pollen was used. Insecticide application, particularly spinosad, determined a reduction in K. aberrans abundance. Pollen applications were associated to an increase in predatory mite population. Significant interaction between pollen applications was observed: pollen application mitigated the effect of chlorpyrifos (because of its compensation of sublethal effects on fecundity) but no effect was observed on spinosad treated plants. In the last work mechanisms of resistance to chlorpyrifos due to mutations in insensitive Acetylcholinesterase (AChE) in K. aberrans was investigated. AChE cDNA was cloned. The cloning strategy relied on the AChE sequence of Tetranychus urticae. To obtain the homologous cDNA sequence in K. aberrans, degenerate primers were designed on conserved functional domains of the annotated AChEs in Metaseiulus occidentalis genome, and combined with 5’-3’ rapid amplification of cDNA ends. When the cloned AChE sequence was compared between K. aberrans sensible and resistant strains, only a Glycine to Serine (G119S) substitution was detected while both AChE sequences had a Phenylalanine residue at 331 position. These results excluded the F331W mutation as responsible for chlorpyrifos target site resistance in K. aberrans but it opened interrogatives about the role of G119S. Indeed the G119S substitution had been already described in mosquitoes Anopheles gambiae and Culex pipiens where it led to AChE1 insensitivity or reduced AChE1 activity. On the contrary, the same aminoacid change in T. urticae AChE sequence was associated with a more moderate decrease in chlorpyrifos response. So a species-specific effect of this substitution cannot be kept out. In any case, site oriented sequencing of AChE cDNA, confirmed the presence of G119S substitution in further three resistant strains of the mite predator as well as the absence of F331W mutation. At the same time, two additional sensible strains, displayed a Glycine at the position 119, instead of Serine, in their AChE sequence. Altogether, these findings might suggest that the G119S polymorphism was involved in the polygenic control of the resistant phenotype. In addition or alternatively, the mutated AChE might be in linkage with one of the genetic determinants which affect the insecticide sensitivity in K. aberrans.

Analisi dell’allergenicità di polline di Citrus clementina e valutazione della cross-reattività con polline di graminacee Rosa Anna Iorioa, Elisabetta Calamellib, Chiara Pulab, Giampaolo Riccib, Donatella Serafini-Fracassinia and Stefano Del Ducaa* aDepartment of Biology E. S., University of Bologna, via Irnerio 42, 40126, Bologna, Italy. stefano.delduca@unibo.it bDepartment of Pediatric Allergology of the Women's, Children's and Adolescents' Health Department of the Gozzadini Children's Hospital Sant'Orsola-Malpighi, University of Bologna, via Massarenti 11, 40138, Bologna, Italy. Il consumo regolare di frutta e verdura è incoraggiato poiché alla base di una dieta giornaliera sana e in grado di migliorare la salute e prevenire o ridurre l'incidenza di varie malattie; sfortunatamente, circa 1/5 della popolazione dei paesi Occidentali soffre di allergie alimentari. L’allergia agli agrumi non è molto frequente come altre allergie alimentari e polliniche (www.foodallergyitalia.org), ma quando presente, spesso è associata a pollinosi grazie ad un fenomeno di cross-reattività, per cui i pollini sarebbero la causa di una sensibilizzazione per via respiratoria che potrebbe predisporre all’allergia verso alimenti che contengono proteine omologhe a quelle sensibilizzanti polliniche. Circa il 10% dei bambini con allergia alimentare mostra sintomi dopo l’ingestione di arance, mentre circa il 17% di pazienti adulti italiani soffre di sindrome orale allergica (SOA) dopo l’ingestione di frutta e verdura. E’ stata documentata l’associazione, in oltre il 90% dei casi, tra sensibilizzazione IgE mediata in pazienti con sintomatologia clinica dopo ingestione di arance e positività dello skin prick test (SPT) per graminacee ed olivo. In questo lavoro i due allergeni maggiori delle arance, Cit s 1 (Germin-like Protein, GLP) e Cit s 2 (profilina), sono risultati correlati alla presenza di sensibilizzazione rispettivamente per il polline di olivo e cipresso (Cit s 1) e per il polline di platano (Cit s 2). La valutazione clinica ha previsto l’identificazione ed il reclutamento di pazienti allergici /sensibilizzati agli agrumi in una popolazione di soggetti con pollinosi (allergia respiratoria) tramite l’anamnesi di allergia alimentare, SPT con estratti commerciali per agrumi e pollini potenzialmente cross-reagenti, nonché Prick by Prick test (PPT) con frutti maturi (polpa). Il siero di alcuni dei pazienti reclutati è stato utilizzato in prove ELISA ed in western blotting dimostrando di avere anticorpi specifici per alcune proteine del polline di Citrus clementina, che in funzione del peso molecolare si potrebbe ipotizzare essere Cit s 1 e Cit s 3 (non-specific Lipid Transfer Protein, nsLTP). Lo studio della reattività crociata con i pollini si è focalizzato sull’analisi dell’espressione dei geni Cit s ben caratterizzati in frutto anche in polline di Citrus c., in modo da valutarne il potenziale allergenico e la cross-reattività con polline di graminacee, in particolare polline di Triticum aestivum. La ricerca di agenti sensibilizzanti in polline è continuata con i) la valutazione dell’espressione di due isoforme della fosfolipasi secretoria (PLAα, PLAβ), enzimi chiave nella sintesi degli eicosanoidi pro-infiammatori; ii) la valutazione dell’attività dell’enzima transglutaminasi (TGasi), che modificando post-traduzionalmente la fosfolipasi ne aumenterebbe l’attività; iii) la capacità dei pollini di produrre specie reattive dell’ossigeno (ROS). E’ quindi ragionevole approfondire la ricerca di questi fattori importante nella patogenesi delle allergie respiratorie in pollini in grado di suscitare sensibilizzazione e cross-allergie agli agrumi.

The study presents a palaeoenvironmental reconstruction based on pollen, no-pollen palynomorphs (= NPPs) and microcharcoals analyses on samples of two cores (PL1 e PL2) taken at Piano Locce ( 1225 m a.s.l., Barisciano, L'Aquila - Italy), within a multidisciplinary programme. Piano Locce is a tectono-karstic depression on the southern side of the “Massiccio del Gran Sasso d’Italia” mountain and it is a hollow that trapped the sediments eroded from the soils of the sorrounding mountains and the wind carried pyroclastic ashes. Methods – Core PL1, drilled in 2003, 80 m long without reaching the limestone at the base, was sampled for pollen analysis following the stratigraphy to obtain an ample vegetational/climatic reconstruction. Core PL2, drilled in 2008, 10 m long, was sampled approximately every 5-10 cm to obtain a detailed reconstruction of the floristic-vegetational/climatic events and of human impact during the Late Pleistocene and the Holocene. For this study 91 of the 260 samples taken from the two cores were studied: 10 from PL1 core (from 0 to 5.14 m) and 81 from PL2 core. The finds analysis, the data exposition and the results interpretation principles followed the current standards. For microcharcoals a particle-count method that divides them in 5 classes was carried out. Chronology – Two datings based on chemical analysis of volcanic minerals are available: core PL1 is dated 170.000-200.000 years BP at the bottom and core PL2 is dated 36.000 years BP at 5 m depth. Results, Discussion, Conclusions – Pollen analysis of Piano Locce series (46.150 pollen finds in the two cores) provides the history of vegetal landscape from 36.000 years BP, plus former data. The finding of high percentage of Artemisia pollen, with Chenopodiaceae and Poaceae and low percentage of arboreal plants suggests, for the lower part of the sequence, a steppe vegetation environment referable to the Last Glacial. In this part some zones may be included in the Last Glacial Maximum (minimum of woody plants pollen), in the interstadial Bolling-Allerod (low Artemisia, relative increase and differentiation of trees/shrubs) and in the Younger Dryas (arboreal plants decrease and Artemisia rise). The transit from the Last Glacial to the Holocene is testified by an arboreal plants increase and the affirmation of mixed Oak-wood (at about 1.50 m depth in PL2). Data suggests that in the area a grass-land grazed quite precociously persisted during the time. Wood always remained far from Piano Locce depression and it was formed alternatively by conifers and broadleaves and subsequently by the predominance of Oak-wood broadleaves. Beech-wood is little represented, probably due to local climatic factors. The combination of Cultivated/cultivable plants begins to be significant in the Medium Holocene and it is obvious in the Upper Holocene. This is testified by Castanea, Juglans, Olea, Vitis e cereals. Prunus dulcis was found in the most superficial samples. NPPs (1.797 finds identified in PL2 core) in the Upper Pleistocene show mainly algal microremains (Pediastrum and Botryococcus colonies and Spirogyra spores) attesting a lacustrine sedimentation, while coprophilus fungi spores appears in good percentage during the Holocene, in a terrestrial environment with periodical water puddles. This fungi presence, like Sporormiella-type, Sordiaria-type, Apiosordiaria and Valsaria-type, in levels caracherized by Cichoriodeae, Asteroideae, Cirsium-type, Galium-type, Ranunculaceae, Stellaria-type pollen suggests the hypothesis that Piano Locce depression was exploited as a grazing area, first by wild animals, then by domestic ones. Microcharcoals (analyzed in PL2 core) are less represented in the lower part of the sequence, maybe due to the presence of humans living in caves and shelters settled in areas at lower quotes than Piano Locce. The presence of bigger sized particles testifies nearer fires, natural or caused by the hunter-gatherers that hunted in the area. During the Holocene the sum of microcharcoals increases, according to the evidence of anthropization provided by pollen and no-pollen palynomorphs.

The study presents the data obtained by palynological and microantrachological analyses of samples taken from the Medieval settlement recently unearthed (excavations directed by P. Pensabene) in the southern area of the renowned Villa del Casale of Piazza Armerina (Enna-Sicily). The aim of the work is to reconstruct the history of the natural and cultural vegetal landscape of the settlement and its sorroundings and to compile, based on pollen flora, a list of plants useful to restore the green setting of the site. Materials and methods – 42 samples from the Medieval settlement and from a related vertical profile were studied, covering a time span from the Byzantine age to modern age, plus a recent control sample (moss sample). Chronology is based on archaeological and somewhere palynological evidence. Results – The samples showed a satisfactory pollen concentration, a sufficient state of preservation and a remarkable floristic diversity (about 236 pollen taxa were identified). On the whole pollen shows an open hilly Mediterranean cultural landscape, largely devoted to meadows and pastures, testified by Cichorioideae, Gramineae and many other herbs (e.g. Aster type, Anthemis type, Leguminosae, Mentha type, Umbelliferae). Human influence is testified by a very strong forest clearance, already occurred in the Byzantine fase. Pollen flora anyway testifies wood remnants from different vegetation belts (e.g. Abies, Betula, Carpinus, Castanea, Fagus, Fraxinus ornus, Myrtus, deciduous Quercus, Q. cf. cerris, Q. cf. virgiliana, Quercus cf. ilex, Pinus cf. halepensis, P. indiff., Phyllirea, Pistacia, Ulmus, Tilia) and fresh water plant communities (Alnus, Butomus umbellatus, Carex, Myriophyllum, Nymphaea, Populus, Sagittaria, Salix, Typha angustifolia,etc.). Besides woody plants suitable for timber, pollen testified a number of useful plants, cultivated or exploited in the wild, for food, decoration or other uses (e.g. Avena-Triticum group, Beta, Corylus, Cynara cf., Hordeum group, Mentha, Nerium oleander, Olea, Platanus, Pinus cf. pinea, Prunus, Secale cereale, Thymus, Vitis ). Some records in the Middle age samples are worth noting: Pistacia vera (pistachios) and Syzygium aromaticum (cloves). Microcharcoals were consistent due to the continuous human activity in the settlement. Conclusion – The pollen history of the site shows a continuity of tradition during the time. Changes occurred, but the different cultures that followed one another during the time seem to have progressively intermingled in the practices and uses concerning plants and plant landscape. The most strong change occurs in the recent moss sample, affected by cultivation of different olive trees (compared with the past ones) and exotic plants ( Eucalyptus).

Mariotti Lippi, M. and Mercuri, A., 1992. Palynology of a resin from an Egyptian coffin of the second century B.C. Rev. Palaeobot. Palynol., 71: 207-218. A palynologicai analysis was carried out on a small amount of resin from the inside of an anthropoid Egyptian stone coffin from the second century B.C. Pinus halepensis Miller is the best represented resiniferous plant (5.3%) in the pollen spectrum and the botanical origin of the resin can be ascribed to it. The only group of pollen which is relatively abundant in the spectrum is Poaceae (29.6%). This high percentage suggests that grasses were in bloom during the period the resin was collected (late spring or summer). Pollen of Quercus ilex, Olea europaea, Ostrya carpil~(folia, Castanea sativa were also found. Pollen grains of Hordeum and Avena/Triticum types together with those of Plantago, Urtica, Chenopodiaceae, Rumex and Arwmisia represent anthropogenic indicators. A phytogeographical interpretation of the found pollen assemblage suggests the eastern part of the Mediterranean as the most probable area v, here the resin was collected. The presence of Cedrus libani pollen suggests an origin from south-eastern Turkey, north-western Syria or northern Lebanon. The flora of this area is in accordance with the pollen assemblage found in the studied resin.

Le proteine allergeniche della mela presentano delle caratteristiche comuni tra loro: - Tutte le proteine allergeniche sono codificate da famiglie geniche composte da un numero variabile di geni, gli isoallergeni, di cui per ognuna esitono numerose varianti alleliche. Questa presenza ridondante di sequenze Mal d nel genoma del melo, ed in particolare su cromosomi omologhi, sembra essere dovuta alla natura allopoliploide del melo e questo fa anche supporre che il numero di isoallergeni noti sia destinato a crescere ulteriormente. - Tre classi di proteine su quattro (Mal d 1, Mal d 2 e Mal d 3) appartengono alle proteine PR e quindi sono in qualche modo correlate alla difesa della pianta da vari tipi di stress, sia biotici che abiotici. - Alcuni fattori comuni a molti allergeni come la stabilità a bassi pH, alle alte temperature e a proteolisi o la capacità di trasportare lipidi o altri ligandi potrebbero influenzarne l’allergenicità. Nonostante l’aumento delle conoscenze in questi ultimi anni, non è stato ancora chiarito quali isoallergeni siano effettivamente espressi nel frutto e quali siano i veri responsabili della reazione allergica. Questo è un punto chiave nel problema delle allergie alla mela ma anche alla frutta e agli ortaggi in generale. Nella impostazione della ricerca su cui abbiamo qui riferito, si è scelto un approccio multidisciplinare che prevede l’integrazione, a fianco delle biotecnologie, di discipline come l’allergologia e la pomologia (Figura 8). Attraverso questi studi si tenterà di associare ad ogni locus genico un determinato potenziale allergenico e quindi, individuando le sequenze di isoallergeni associate ad un basso grado di allergenicità, si potrà anche orientare il miglioramento genetico del melo verso l’ottenimento di varietà a bassa allergenicità. Una volta individuati dei marker per la bassa allergenicità, si potranno cercare genotipi interessanti per questo carattere anche esplorando le collezioni dell’antico germoplasma del melo. Un aspetto però da non sottovalutare è la doppia faccia della medaglia ovvero il ruolo delle proteine responsabili dell’allergenicità; molte di queste, infatti, sono coinvolte nella risposta della pianta ad attacchi di patogeni e altri stress. Questa considerazione rende inevitabile la preoccupazione che l’auspicata riduzione dell’allergenicità, possa comportare una maggiore suscettibilità agli stress indesiderabile per ovvi motivi. Per ora è solamente un interrogativo. Considerando i notevoli effetti benefici delle mele sulla salute degli individui e considerando il notevole interesse che, negli ultimi anni, il mercato riserva alla nutraceutica e agli alimenti correlati al benessere, non è da escludere che lo studio del carattere riguardante l’ipoallergenicità dei frutti possa essere interessante non solo per quanto riguarda la salute umana ma anche per un possibile riscontro economico