Effect of production process and high-pressure processing on viability of Listeria innocua in traditional Italian dry-cured coppa

In this study the effect of the application of High Pressure Treatment (HPP) combined with four different manufacturing processes on the inactivation of Listeria innocua, used as a surrogate for L. monocytogenes, in artificially contaminated coppa samples was evaluated in order to verify the most suitable strategy to meet the Listeria inactivation requirements needed for the exportation of dry-cured meat in the U.S. Fresh anatomical cuts intended for coppa production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with Listeria innocua using a mixture of 5 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing Listeria spp. The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14\ub0C. Listeria innocua was enumerated on surface and deep samples post contamination, resting, ripening and HPP treatment. The results of this study show how the reduction of the microbial load on coppa during the production process did not vary among three companies (P>0.05) ranging from 3.73 to 4.30 log CFU/g, while it was significantly different (P<0.01) for the fourth company (0.92 log CFU/g). HPP treatment resulted in a significant (P<0.01) deep decrease of L. innocua count with values ranging between 1.63-3.54 log CFU/g with no significant differences between companies. Regarding superficial contamination, HPP treatment resulted significant (P<0.01) only in Coppa produced by two companies. The results highlight that there were processes less effective to inhibit the pathogen; in particular for company D an increase of L. innocua count was shown during processing and HPP alone cannot be able to in reaching the Listeria inactivation requirements needed for exportation of dry-cured meat in the U.S. According to the data reported in this paper, HPP treatment increases the ability of the manufacturing process of coppa in reducing Listeria count with the objective of a lethality treatment

Effect of production process and high-pressure processing on viability of Salmonella spp. in traditional Italian dry-cured coppa

The aim of the study was to investigate the combined effect of the manufacturing process followed by HPP treatment on the inactivation of Salmonella spp. in artificially contaminated coppa samples, in order to verify the ability of the combined processes to achieve the objective of a 5-log reduction of Salmonella spp. needed for exportation to the U.S. Fresh anatomical cuts intended for coppa production were supplied by four different delicatessen factories located in Northern Italy. Raw meat underwent experimental contamination with Salmonella spp. using a mixture of 3 strains. Surface contamination of the fresh anatomical cuts was carried out by immersion into inoculum containing Salmonella spp. The conditions of the HPP treatment were: pressure 593 MPa, time 290 seconds, water treatment temperature 14\ub0C. Surface and deep samples were performed post contamination (T0), end of the cold phase (T1), end of process (Tend), and after HPP treatment (postHPP) and Salmonella spp. Enumerated. The results of this study show a significant reduction of Salmonella spp. all through the production process (P<0.01) for all companies, followed by an additional reduction of bacterial counts due to HPP treatment (P<0.01), both in superficial and deep contaminations (P<0.01). The superficial overall reduction resulted of 1.58 to 5.04 log CFU/g during the production process. HPP treatment resulted in a significant (P<0.01) superficial and deep decrease in Salmonella spp. enumeration varying from 0.61 to 4.01 log and from 1.49 to 4.13 log. According to the data presented in this study, only the combined approach of coppa manufacturing process followed by HPP treatment always led to a 5-log reduction of Salmonella spp. required by USDA/FSIS guidelines

MOLECULAR IDENTIFICATION OF VENTURIA ASPERATA FROM ATYPICAL SCAB-LIKE SYMPTOMS ON APPLES IN ITALY

Similar apple scab symptoms were first observed at the end of July in 2012 in the Northern Italy (Cesena) on fruits of apple cultivar Modì carrying the Rvi6 major resistance gene to Venturia inaequalis. The aim of this work was to identify the causal agent of the atypical scab-like symptoms by molecular techniques. Symptomatic fruits were collected during May in one orchard in 2015. Ten monoconidial isolates were obtained through recovering the conidia from about 10 fruits. Conidial suspension was then streaked on Petri dishes of water agar amended with streptomycin sulfate. After 24 h of incubation at 20°C, single germinated spores were selected under stereomicroscope, then picked up and placed on PDA amended with three antibiotics. The isolates were cultivated at 20°C until molecular characterization together with the reference strain of Venturia asperata. Amplification of ITS fragments was carried out for specifically amplify rDNA of V. asperata, V. inaequalis and Venturia pirina. Approximately 4-5 hyphae were removed from each isolate and transferred without DNA extraction to the PCR tube with the addition of BSA. DNA amplification was obtained for all isolates by primers specific for V. asperata, while no amplification was observed using primers specific for V. inaequalis and V. pirina. These results point out the presence of V. asperata from the atypical scab-like symptoms but further studies are in progress to have a more precise identification of the pathogen

Control strategies for Colletotrichum acutatum on strawberries in North Italy

Studies were conducted during three fruiting seasons on anthracnose susceptible cv \u2018Onda\u2019 (first year) and cv \u2018Alba\u2019 (second and third year) strawberry cultivars in the Romagna Region of Italy. In the spring, four and two weeks before bloom, the plants were inoculated by spraying each plant with a 10 ml-volume of a C. acutatum conidial suspension (106 conidia ml-1). To test the in vivo efficacy of a newer fungicide compared with older ones used in conventional spray programs the following fungicides were used: a combination of cyprodinil (37.5% a.i.) and fludioxonil (25% a.i.) (trade name Switch) at 0.8 kg ha-1; a combination of boscalid (26.7% a.i.) and pyraclostrobin (6.7% a.i.) (trade name Signum) at 1.8 kg ha-1; azoxystrobin (23.2% a.i.) (trade name Ortiva) at 0.9 l ha-1. Two treatments were applied: the first a week before full-bloom and the second at the full-bloom stage. The timing of Signum fungicide application was also evaluated in the same fields but in an other batch of fruits inoculated with C. acutatum as described above. Berries were harvested in threefold, examined and categorized as marketable or diseased fruit. Marketable fruit was stored for 2 days at 2\ub0C and another 3 days at 20\ub0C and examined at each time interval. Treatments with pyraclostrobin+boscalid or cyprodinil+fludioxonil lead to higher marketable yields, significantly reducing the infections with respect to controls in all years, with an efficacy ranging from 50 % to 81%. Azoxystrobin reduced the infections significantly with respect to controls but less effectively than pyraclostrobin+ boscalid and cyprodinil+fludioxonil. Two applications of Signum during bloom were able to control anthracnose, although only one treatment near harvest may be useful when wheatear conditions are favourable to C. acutatum development

Ten years of field trials on grey mold control on strawberries

The region of Romagna (north-central Italy) is one of Italy\u2019s most typical areas for growing strawberries, with over 400 hectares of cultivated surface area. Over two-thirds of this crop consists of strawberry beds in open fields in which the onset of phyto-sanitary problems depends above all on climatic trends. In this context the development and diffusion of various pathogenic agents is more likely; these agents can lead to consistent product losses in the field as well as during the post-harvest period. The most common pathogen is Botrytis cinerea, a grey mold agent with ubiquitous inoculation in the areas where strawberries are commonly grown. The research reported is a synthesis of ten years (1998-2007) of field experimentation, aimed at evaluating the efficiency of various active ingredients used against B. cinerea. During the ten year period of experimentation the older generation of active ingredients was evaluated (procimidone) as well as more recent ones (pyrimethanil, mepanipirym, fenhexamide, cyprodinil-fludioxonil, and azoxystrobin), and finally the most recent phyto-iatric acquisitions (boscalid-pyraclostrobin). The most effective fungicides for controlling strawberry grey mold and limiting financial loss in all phases were pyrimethanil, cyprodinil-fludioxonil, mepanipirym and boscalid-pyraclostrobin (average effectiveness 85-95%). fenhexamide and procimidone were less effective than the aforementioned ones (average effectiveness 70-80 %) and finally azoxystrobin was modestly effective (50-60 %)

Verifiche sull'attivit\ue0 di ziram e dodina contro la bolla del pesco in Emilia Romagna

RIASSUNTO A seguito della segnalazione di non sempre soddisfacenti prestazioni della dodina nella lotta alla bolla del pesco in Emilia-Romagna, sono state condotte in tre aree della regione dal 2004 al 2009 prove parcellari per verificare l\u2019attivit\ue0 del fungicida a confronto con ziram, che come la stessa dodina \ue8 da parecchi anni un prodotto di riferimento nella difesa dalla malattia. Si \ue8 operato eseguendo i trattamenti nelle epoche tradizionali (tra la caduta delle foglie e la fioritura), valutando la protezione a livello di germogli e foglie. Le prove hanno evidenziato un diverso comportamento dei due prodotti: efficacia della dodina costantemente elevata e quasi sempre superiore rispetto a quella dello ziram nelle verifiche condotte a Bologna (in presenza di una elevata pressione infettiva); risultati opposti in quelle di Ravenna e Forl\uec in cui (in presenza di una pressione media) la dodina ha controllato solo parzialmente la malattia, mentre lo ziram ha sempre mostrato una soddisfacente protezione. Le cause della diversa risposta dei due prodotti nelle prove condotte nelle due aree non sono facilmente spiegabili. Se l\u2019insoddisfacente protezione da parte dello ziram a Bologna pu\uf2 essere giustificata dalla elevata pressione infettiva, quella altrettanto scarsa della dodina potrebbe essere collegata o a un dosaggio non sempre pieno (es. a Ravenna) oppure a una eventuale riduzione di sensibilit\ue0 di Taphrina deformans nell\u2019aree di Ravenna e Forl\uec, dove lo stesso prodotto \ue8 stato a lungo diffusamente utilizzato. SUMMARY COMPARISON OF THE ACTIVITY OF ZIRAM AND DODINE IN CONTROL OF PEACH LEAF CURL (TAPHRINA DEFORMANS) For about 20 years dodine has been successfully used in Italy to control peach leaf curl (Taphrina deformans (Berk.) Tul.) but cases of unsatisfactory field activity were recently reported in Emilia Romagna region. Therefore field plot trials were carried out to test the activity of dodine in comparison with ziram, the most used fungicide in peach protection from T. deformans. The trials were conducted in peach orchards over six years in Bologna and Ravenna/Forl\uec areas, applying the two products 2-4 times between leaf fall and blooming. Dodine and ziram showed different protection in the two considered areas: the first performed very well in Bologna trials (with high disease pressure) and almost always better than ziram, whereas in Ravenna and Forl\uec ones better control was always exerted by ziram while dodine showed an unsatisfactory activity. The poor control performed by ziram may be explained by the high disease severity whereas the poor results of dodine may be related to a low application rate or to its possible reduction of activity against T. deformans because of intensive use of this fungicide after its introduction in peach leaf curl control in early \u2018nineties

Approccio anatomopatologico di uno schwannoma maligno in Carassius auratus

Gli schwannomi sono neoplasie derivanti dalle guaine dei nervi periferici. Nelle specie ittiche sono stati descritti in pesce rosso (Carassius auratus), dentice (famiglia Lutianidae), salmone argentato (Oncorhynchus kisutch), bicolor damselfish (Pomacentrus partitus) e sperlano americano (Osmerus mordax). Generalmente queste neoplasie hanno un comportamento biologico benigno; tuttavia rari casi di schwannoma maligno sono stati descritti in pesce persico (Perca fluviatilis) e salmone argentato (Oncorhynchus kisutch). L'elevata incidenza degli schwannomi in alcune popolazioni ittiche ha suggerito una possibile eziologia virale, confermata solo nel damselfish. La distinzione tra schwannoma e neurofibroma in patologia animale e umana rimane ad oggi ancora un problema diagnostico e l\u2019approccio ultrastrutturale \ue8 stato indicato come strumento d\u2019elezione per differenziare le due neoplasie. Un pesce rosso adulto di 11 cm di lunghezza presentava una massa multinodulare esofitica bianco-rosata nella regione dorsale che coinvolgeva la pinna dorsale; il soggetto \ue8 stato sottoposto ad indagini ultrasonografiche che hanno evidenziato un\u2019area a margini netti di forma ovalare ad ecostruttura parenchimatosa omogenea delle dimensioni di 22 x19x17 mm. L\u2019esame color Doppler ha evidenziato una intensa vascolarizzazione intralesionale. Dopo sedazione ottenuta tramite immersione del soggetto in soluzione acquosa con anestetico MS-222, \ue8 stato eseguito esame citologico della massa. Nei preparati colorati con May Gr\ufcnwald-Giemsa si osservavano numerose cellule mesenchimali moderatamente pleomorfe sia isolate che raggruppate in piccoli ammassi irregolari, frammiste a una modica quantit\ue0 di matrice extracellulare amorfa. Tali cellule, di forma da ovale a fusata con estremit\ue0 allungate e dimensioni variabili da 70 a 200 \ub5m, avevano elevato rapporto nucleo:citoplasma, limiti cellulari indistinti e scarso citoplasma debolmente basofilo, talora microvacuolizzato. I nuclei, centrali e di forma da ovale a sigariforme, mostravano cromatina granulare con nucleoli spesso multipli. Sul fondo dei preparati erano presenti numerosi eritrociti, leucociti di derivazione ematica, brandelli di capillari e detriti cellulari. Il sospetto citologico di neoplasia mesenchimale maligna, le notevoli dimensioni della massa e le caratteristiche ecografiche suggestive di marcata angiogenesi e potenziale rapido accrescimento, hanno fatto propendere per l\u2019eutanasia del soggetto. Successivamente, la massa \ue8 stata sottoposta ad approfondimento istologico ed ultrastrutturale. All\u2019istologia la neoplasia si mostrava non incapsulata e rivestita da un sottile epitelio di derivazione cutanea; era caratterizzata da una ricca vascolarizzazione con presenza di aree emorragiche e macrofagi carichi di emosiderina. Il tessuto neoplastico era intensamente cellulare e composto da cellule fusate con nuclei a palizzata disposte in fasci paralleli o in spirali secondo il tipo Antoni A e intercalate da tessuto connettivo. Numerose cellule neoplastiche presentavano segni di atipia cellulare come la frequente presenza di nucleoli multipli. Le figure mitotiche erano rare. Il tessuto neoplastico mostrava un comportamento localmente invasivo andando ad infiltrare profondamente il muscolo scheletrico della regione dorsale. L\u2019ultrastruttura ha messo in evidenza alcune caratteristiche morfologiche analoghe a quelle descritte negli schwannomi maligni. In conclusione, si tratta della prima descrizione citologica corredata di indagine ecografica di uno schwannoma in una specie ittica; la forte concordanza fra reperti citologici e istologici suggerisce l\u2019impiego costante della metodica citologica per finalit\ue0 diagnostiche anche in queste specie