Molecular diagnosis of Bursaphelenchus xylophilus (Steiner & Buhrer) Nickle (Nematoda Aphelenchida Parasitaphelenchidae) in presence of polyphenols and other inhibitory compounds.

The pine wood nematode Bursaphelenchus xylophilus (Aphelenchida Parasitaphelenchidae) infects conifers, in particular pines, causing pine wilt disease. This nematode is considered a quarantine organism according to the EPPO and AQISQ guidelines. For this reason, several procedures for an affordable diagnostic test have been developed based on LAMP technique. In this work we shown how this technique behaved in presence of compounds that exert an inhibitory effect on DNA polymerases, a common problem that could be found in DNA preparations obtained from samples included in plant matrixes  LAMP technique was found able to detect presence of its specific DNA target in almost all cases studied but we found that the addiction of polyphenolic compounds present in some woody matrixes to DNA preparations could affect significantly on the sensibility of diagnostic test, while resinous material had a lesser influence on amplification

BIOLOGICAL CONTROL OF ACLEES SP. CF. FOVEATUS AND FIRST RECOVERY OF AN ASSOCIATE BEAUVERIA BASSIANA STRAIN

Aclees sp. cf. foveatus (Coleoptera: Curculionidae) is spreading in Central Italy, causing severe infestation on fig trees. There are very few information for this pest and no natural enemy is reported. Here, we report the first recovery of a natural enemy associated with this invasive weevil, a strain of the entomopathogenic fungus Beauveria bassiana. The potential use of entomopathogenic fungi and nematodes as biocontrol agents was tested against adults in laboratory trials. In agree with the detection in nature, only treatments with B. bassiana were able to control the insects. This result opens new frontiers for the environmental friendly control strategies against this weevil

Effects of cryopreservation and freeze-drying on proteases enzymatic activity of entomopathogenic strains of Beauveria bassiana (Balsamo) Vuillemin and Metarhizium anisopliae (Metchnikoff) Sorokin.

Beauveria bassiana and Metarhizium anisopliae are fungal species known for their entomopathogenic activity; they represent an important alternative to chemical control of many pest insects and appropriate preservation methods are thus necessary to maintain their properties over time. In the present study the effects of cryopreservation and freeze­drying were investigated on virulence determinants such as proteases and specifically on subtilisin-type Pr 1 and Pr 2 that seem to play a predominant role in insect fungal penetration. Three Metharizium anisopliae strains (M 13/ I05, M 13/ I12 and M 13/ I33) and three Beauveria bassiana strains (B 13/ I03, B 13/ I57 and B 13/ I63) were tested. Data obtained from our experiments show that the decrease of Pr 1 and Pr 2 enzymatic activity in B. bassiana and M. anisopliae strains 7 months after storage does not affect their entomopathogenic ability against larvae of Tenebrio molitor compared to the fresh cultures before storage. The role of Pr 1 and Pr 2 as predominant factors in the entomopathogenicity of both fungal species is discussed. No significant differences were found in any strain tested for morphological features using both preservation methods

A new aphid subspecies on the endemic Cyprus cedar Cedrus brevifolia: Cinara cedri brevifoliae ssp.n. (Aphididae Lachninae)

A subspecies of Cinara cedri Mimeur, C. cedri brevifoliae ssp.n., is described from apterous viviparous females. The authors report on the discovery of this new taxon in Cyprus on the endemic cedar Cedrus brevifolia (Hooker fil.) Henry, suggesting that the aphid is co-endemic with its host conifer. Morphological evaluation is provided in order to support this conclusion and a key to separate the Cedar Cinara species is given. Moreover, new insights on the distribution of C. cedri in the Mediterranean area were gained by molecular analysis. Gene sequences were deposited in Genbank and the type specimens located in the collection (A.B.) of CREA-Research Center for Agrobiology and Pedology, Florence, Italy

EVALUATION OF SHEEP GRAZING EFFECTS ON NEMATODE COMMUNITY, INSECT INFESTATION AND SOIL FERTILITY IN SWEET CHESTNUT ORCHARDS: A CASE OF STUDY

In sweet chestnut cultivation, important ecosystem in Europe, the moderate sheep grazing practice may have some beneficial effects in organic farming. Its impact, on soil fertility, soil nematode community and level of fruit infested by tortricids and curculionids was evaluated in a case of study in Tuscany (Central Italy). An organic farm with sheep grazing system (SG) based on long-term field application was compared with an adjacent organic farm with poultry manure system (PM). In both managements, no treatment to control pests and diseases was applied and 0.19 kg /plant of nitrogen were distributed. SG increased twice the amount of organic C, total N and available P than PM. Moreover, the salinity effect of uric acid significantly increased the pH values in SG (from 5.0 to 6.2). The greatest accumulation of organic matter under SG provided stability in nematode community structure and the plant parasitic nematodes decreased. The analysis on the nematodes emerged from Galleria mellonella larvae, revealed that the abundance of native entomopathogenic nematodes was low and not effective to control insect pests. The sheep demonstrated to be the effective predators of tortricid and weevil larvae and in November, the total infestation was significantly lower in SG (34.3%) than PM (54.3%)

Thaumatotibia leucotreta (Meyrick) (Lepidoptera Tortricidae) intercepted for the first time in Italy

In June 2014, Thaumatotibia leucotreta (Meyrick) (Lepidoptera Tortricidae) was found during the inspection of the Italian Phytosanitary Service of Tuscany Region in the port of Leghorn, in a container from South Africa containing Navel oranges. The degree of damage is high, reaching the 2% of the harvested fruits. This is the first interception of this polyphagous pest for Italy

Oscheius tipulae in Italy: Evidence of an Alien Isolate in the Integral Natural Reserve of Montecristo Island (Tuscany)

Montecristo Island is an integral natural reserve of the Tuscan Archipelago National Park (Central Italy), characterized by a peculiar assemblage of flora and fauna, with several endemic taxa, and also with a high number of alien species. During a soil survey, we found an alien Oscheius tipulae Lam &Webster, 1971 isolate, phylogenetically close to others from South America. In this article, we examined the possible pathways of introduction of this nematode. Because of the high number of alien plants in this protected area and the low desiccation survival ability of O. tipulae, we hypothesized that the presence of this alien nematode isolate may be related to the soil of introduced plants, although historical association with plant-associated invertebrates is also possible. Further studies with more populations and marker molecules are necessary to investigate the distribution of O. tipulae and the possible impact on this natural reserve

FIRST RECORD OF THE PEST ARTONA (FUSCARTONA) MARTINI EFETOV, 1997 (LEPIDOPTERA ZYGAENIDAE PROCRIDINAE ARTONINI)IN EUROPEAN TERRITORY

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Stoccaggio di uova ospiti a basse e ultrabasse temperature e studi preliminari sulla crioconservazione degli Imenotteri ooparassitoidi

Artropodi e Nematodi comprendono una quota preponderante del numero di specie animali del nostro pianeta e gli studi svolti nell’ambito del Dottorato si inseriscono in un quadro generale di ricerche sulla crioconservazione, avviate a partire dal 2005 presso il Centro di Ricerca per l’Agrobiologia e la Pedologia di Firenze del CRA, mirate alla definizione di materiali e procedure di permeabilizzazione specie-specifiche e all’identificazione degli stadi ottimali per trattamenti di vitrificazione di organismi inclusi in questi due Phyla, con il fine ultimo di realizzare le condizioni per la creazione di “banche genetiche crioconservate”, nelle quali stoccare a lungo termine specie di interesse agroforestale e ambientale (Irdani et al. 2006; Cosi et al. 2008; Irdani et al. 2011; Roversi et al. 2013). Le indagini di cui si riferisce hanno riguardato 2 sistemi modello: 1. Fitofago ospite: Leptoglossus occidentalis Heidemann (Heteroptera Coreidae) • Ooparassitoide: Gryon pennsylvanicum (Ashmead) (Hymenoptera Platygastridae) 2. Fitofago ospite: Galleria mellonella (L.) (Lepidoptera Pyralidae) • Ooparassitoide: Trichogramma brassicae (Bezdenko) (Hymenoptera Trichogrammatidae). Nel primo sistema, dopo la caratterizzazione molecolare del ceppo di Gryon pennsylvanicum (Ashmead), introdotto in Europa dal Canada nell’ambito di un programma di “lotta biologica classica” (Roversi et al. 2011), con riferimento alle due problematiche sopracitate, sono state valutate sia le potenzialità di stoccaggio delle uova di Leptoglossus occidentalis per il successivo utilizzo per la parassitizzazione da parte del Platigastride che le possibilità di permeabilizzazione del corion delle uova del Coreide, quale primo passo per la messa a punto di un protocollo di crioconservazione per uova di ospiti contenenti embrioni di Imenotteri endoparassitoidi, attualmente non disponibile per nessuna specie di questo gruppo. Nel secondo sistema è stata verificata la possibilità di applicare la modalità di permeabilizzazione inserita nel protocollo di crioconservazione messo a punto per gli embrioni di Galleria mellonella (L.) (Roversi et al. 2008), per il trattamento di uova di questo lepidottero contenenti embrioni dell’ooparassitoide Trichogramma brassicae (Bezdenko)