Characterization and pathogenicity assessment of Plectosphaerella species associated with stunting disease on tomato and pepper crops in Italy

This study follows a survey carried out in 2012 and 2013 on tomato and pepper crops in the Foggia province (southern Italy), for morphological, molecular and pathogenicity analyses of Plectosphaerella fungi. The Plectosphaerella genus includes several species that are pathogens of horticultural plants. The survey identified tomato and pepper crops that showed abundant wilt, leaf yellowing, and discolouration and necrosis of roots, plus collar and stem symptoms. Different fungi including Plectosphaerella spp. were isolated from tissues with and without symptoms. Subsequent molecular and morphological studies identified first records of P. citrulli infecting tomato and pepper, and P. pauciseptata and P. ramiseptata infecting pepper. Pathogenicity testing confirmed that most isolated species of Plectosphaerella caused symptoms on tomato and pepper, with P. ramiseptata the most aggressive. On the basis of these data, it is considered that Plectosphaerella species may cause stunting disease in tomato and pepper

Characterization and pathogenicity of Plectosphaerella spp. collected from basil and parsley in Italy

From 2012–2016 plants of basil (Ocimum basilicum L.) and parsley (Petroselinum sativum Hoffm.) showing decline symptoms were collected from local markets in Foggia Province (southern Italy) and assessed to determine their main fungal pathogens. These plants showed symptoms including leaf yellowing, necrotic lesions on stems, collar and roots, and in some cases, stunting of entire plants. Mycological analyses revealed fungal isolates mainly belonging to the Plectosphaerella genus. Molecular and morphological studies identified four species of Plectosphaerella: Plectosphaerella cucumerina, P. pauciseptata, P. plurivora, and P. ramiseptata. To understand the pathogenic roles of these fungi, and five other reference Plectosphaerella spp., pathogenicity tests were performed in vitro and in-vivo using, respectively, detached leaves and 30-d-old plants of basil (cv. Napoletano) and parsley (cv. Gigante di Napoli). All the fungal species isolated produced host symptoms, including necrotic spots, parenchymatic patches, hydropic areas, collar and root discolouration on leaves and young plants, with varying severity. The most aggressive species on both plants were P. pauciseptata and P. ramiseptata, while P. alismatis, P. citrulli, P. cucumerina, P. delsorboi, P. melonis, and P. plurivora gave less disease severity on both plants. This is the first report worldwide of P. cucumerina, P. pauciseptata, and P. ramiseptata as pathogens of parsley, and P. pauciseptata, P. plurivora, and P. ramiseptata as pathogens of basil

Comparison of the performance of carcinogenic HPV typing of the Roche Linear Array and Qiagen LiquiChip® HPV assays

BACKGROUND: Cervical cancer is caused by high-risk types of human papillomavirus (HPV). DNA testing of such high-risk types of HPV could improve cervical screening.The aim of the study was to compare the sensitivities and positive predictive values of two commercially available typing assays (Qiagen LQ and Roche LA) and to comparatively assess the distribution of HPV types with these two assays. METHODS: The study population comprised 311 ASCUS + women with abnormal pap tests who were HCII positive and who were admitted to three European referral gynecology clinics between 2007 and 2010 (Madrid, Marseille and Milan). All patients underwent LQ and LA tests. RESULTS: The sensitivity of the two assays for HPV typing was 94% for LQ and 99% for LA (compared with HCII). The overall concordance between LQ and LA was 93%. The three prevalent genotypes, HPV16, HPV18, and HPV31, were identified with a high concordance using the two assays: kappa 0.93, 0.83, and 0.91, respectively. Mixed genotypes were more frequently detected by LA than by LQ: 52% vs. 18%, respectively (p < .0001). CONCLUSIONS: These assays have a good clinical sensitivity for detecting HPV types in CIN2+ patients and allow the virus type to be detected in the same experiment. Our study revealed no significant difference between LQ and LA for CIN2+ or CIN3+ diagnosis, indicating similar distributions of HPV types and a mixed genotype detection that is higher for LA than for LQ

Occurrence fungi causing black foot on young grapevines and nursery rootstock plants in Italy

Summary. Young grapevine plants with decline and wood necrosis symptoms were collected from vineyards and nurseries in the Apulia and Molise regions, Italy, from 2013 to 2015. Isolations of fungi were prepared from 45 diseased grapevine plants, and the cultures were identified. Several species commonly associated with Petri disease, Botryosphaeria dieback, and black foot disease were isolated. A detailed study was carried out, and 182 isolates resembling Cylindrocarpon-like asexual forms were identified through morphological characterisation and DNA analysis of internal transcribed spacer regions 1 and 2 of the rRNA gene and the partial β-tubulin gene. Dactylonectria torrensensis and Ilyonectria liriodendri were identified based on morphological features and the partial histone 3 gene, so these fungi can be defined as the causal agents of black foot on grapevine for the first time in Italy. Thelonectria blackeriella is also described as a new species, through morphological characterisation and multigenic analysis using sequence data for five loci (large subunit RNA, internal transcribed spacers, β-tubulin, actin, RNA polymerase II subunit 1). This new species was associated with black foot symptoms according to preliminary pathogenicity tests, with representative isolates of each of the three species. Pathogenicity tests showed that these species can cause black streaking in the wood of 1-year-old grapevine rootstock shoots. The identification of D. torresensis, I. liriodendri and T. blackeriella from young grapevine plants and rooted rootstock highlights the importance of black foot disease in Italy, which has previously been overlooked

Microbial diversity of traditional Sicilian cheeses

Traditional Sicilian cheeses are manufactured in small size farms with raw milk from animals of indigenous breeds and without the addition of starter cultures. In order to transform milk into cheese, the presence of lactic acid bacteria (LAB) is required. The main sources of desirable LAB are generally the milk, the rennet, the equipment used during processing and the dairy environment. In the last years, the microbial characterisation of traditional Sicilian cheeses, such as Caciocavallo Palermitano, Protected Designation of Origin (PDO) Pecorino Siciliano and PDO Vastedda della valle del Belìce have been the object of different studies conducted by our research group. To this purpose, the aim of the present study was to describe the microbial population of traditional Sicilian cheeses. The analysis of the microbial diversity of these cheeses revealed the presence of several dairy LAB at dominant levels, while the undesired microorganisms, including coliforms and coagulase-positive staphylococci (CPS), were at very low densities. Overall, the presence of the pathogenic bacteria Listeria monocytogenes and Salmonella spp. was never detected. This may be due to the quality of milk, the optimal maintenance of wooden vats and on following good production practices. Furthermore, the technological characterisation of the LAB found in these cheeses and in raw materials showed interesting dairy properties, including acidification capacity, diacetyl formation, autolytic properties and the ability to inhibit undesired bacteria

Evolution of the lactic acid bacterial biofilms on different wood typologies applied for the production of vats intended for cheese making

The present work was carried out to evaluate the lactic acid bacterial biofilms formation on wooden vat produced with seven different woods, derived from trees species grown in Sicily. The biofilm formation on the wooden surfaces was performed as reported by Gaglio et al. Microbiological and scanning electron microscopy analyses did not show differences in terms of microbial levels and composition within the neoformed biofilms (Fig.1). The specific investigation of Salmonella spp., Listeria monocytogenes, Escherichia coli and positive-coagulase staphylococci did not generate any colony for all vats before and after microbial activation. Lactic acid bacteria (LAB) populations dominated the surfaces of all vats and the highest concentration of approximately 7.63 Log CFU/cm2 was registered for thermophilic cocci. All colonies of different morphologies were isolated and characterized for their physiological and biochemical characteristics before being investigated genetically at strain level by RAPD-PCR and species level by a polyphasic approach consisting of 16S rRNA gene sequencing. As reported in a preview investigation conducted on wooden vat surfaces. we found species within the genera Enterococcus, Lactobacillus, Lactococcus, and Streptococcus. The species most frequently present were Lactobacillus fermentum and Lactococcus lactis. The technological characterization of the LAB found at high numbers on to the surfaces of the wooden vats showed interesting dairy properties. In order to deeply investigate on the safety of the wooden vat, a test of artificial contamination on new Calabrian chestnut vats was carried out showing that the acidity and the bacteriocin generated by LAB represent efficient barriers to their adhesion. This study highlights the importance to use the wooden vats for traditional cheese production and provides evidences for their safety in terms of bacterial community evolution

Formation and Characterization of Early Bacterial Biofilms on Different Wood Typologies Applied in Dairy Production

The main hypothesis of this work was that Sicilian forestry resources are suitable for the production of equipment to be used in cheese making and indigenous milk lactic acid bacteria (LAB) are able to develop stable biofilms providing starter and nonstarter cultures necessary for curd fermentation and cheese ripening, respectively. Hence, the present work was carried out with deproteinized whey to evaluate LAB biofilm formation on different woods derived from tree species grown in Sicily. Microbiological and scanning electron microscopy analyses showed minimal differences in microbial levels and compositions for the neoformed biofilms. The specific investigation of Salmonella spp., Listeria monocytogenes, Escherichia coli, coagulase-positive staphylococci (CPS), and sulfite-reducing anaerobes did not generate any colony for all vats before and after bacterial adhesion. LAB populations dominated all vat surfaces. The highest levels (7.63 log CFU/cm2) were registered for thermophilic cocci. Different colonies were characterized physiologically, biochemically, and genetically (at strain and species levels). Six species within the genera Enterococcus, Lactobacillus, Lactococcus, and Streptococcus were identified. The species most frequently present were Lactobacillus fermentum and Lactococcus lactis. LAB found on the surfaces of the wooden vats in this study showed interesting characteristics important for dairy manufacture. To thoroughly investigate the safety of the wooden vat, a test of artificial contamination on new Calabrian chestnut (control wood) vats was carried out. The results showed that LAB represent efficient barriers to the adhesion of the main dairy pathogens, probably due to their acidity and bacteriocin generation. IMPORTANCE This study highlights the importance of using wooden vats for traditional cheese production and provides evidence for the valorization the Sicilian forest wood resources via the production of dairy equipment

Management of Branched Broomrape in Field Processing Tomato Crop

In recent years, there has been a considerable increase in land area used for tomato (Lycopersicon esculentum Mill.) in many countries around the world. The essential role is played by Italy at a worldwide level as the country with the third biggest production of tomatoes for processing. Phelipanche ramosa (L) Pomel, commonly known as branched broomrape, is a root holoparasitic weed for many crops, particularly for the processing tomato. Due to its physical and metabolic overlap with the crop, its underground parasitism, and hardly destructible seed bank, the control of this parasite in the field is difficult. Results of research studies, many of them on environmental-friendly methods such as preventive, agronomic, and biological carried out in southern Italy, are discussed and summarized. The results can constitute a relevant basis for further experimental studies

A long-term posttraumatic study in the rat median nerve crush injury model

The possibility that posttraumatic regeneration may eventually lead rat peripheral nerve fibers back to normal is still under debate. While it has been shown that, after a sufficiently long posttraumatic time, the number of regenerated rat peripheral nerve fibers can return to normal levels and the animal can regain normal pre-lesion function, no information regarding long-term changes in size parameters of regenerated nerve fibers is still available. To fill this gap, 24-week posttraumatic changes in myelinated axon and nerve fiber diameter, myelin thickness and g-ratio (axon diameter/fiber diameter), distal to a nerve crush (axonotmesis lesion) of the rat median nerve were assessed by stereology. Results showed that, while as expected fiber number returned to normal values at week-24, both axon and fiber diameter and myelin thickness were still significantly lower at week-24 in comparison to pre-lesion values. Finally, g-ratio, which persisted unmodified along the regeneration process, eventually resulted to be significantly reduced at week-24 in comparison to pre-lesion value. Based on these results, the hypothesis that regenerated rat peripheral nerve fibers are able to spontaneously return to normal, provided that a sufficiently long time recovery post-axonotmesis is allowed, is rejected

Polyphasic characterization of microbiota of “Mastredda”, a traditional wooden tool used during the production of PDO Provola dei Nebrodi cheese

11openInternationalItalian coauthor/editorThe biofilms of the wooden tables used for the acidification of the curd were investigated for PDO Provola dei Nebrodi cheese, a traditional stretched cheese made in eastern Sicily (southern Italy) from raw cows’ milk. To this purpose the wooden tables of four dairy facilities were analysed for their microbiota by scanning electron microscopy (SEM) analysis and a combined culture-independent and -dependent microbiological approach. SEM inspection showed an almost continuous biofilm formation. MiSeq Illumina analysis identified 8 phyla, 16 classes, 25 orders, 47 families and 50 genera. Corynebacterium, Bifidobacterium and lactic acid bacteria (LAB) were detected in all samples. In particular, the LAB genera detected on all wooden tables were Lactobacillus, Streptococcus and Lactococcus. LAB dominated the surfaces of all wooden tables with levels higher than 7.0 Log CFU/cm2. In particular, the LAB found at the highest levels were mesophilic cocci. Coagulase positive staphylococci, Salmonella spp., Listeria monocytogenes and Shiga-toxigenic Escherichia coli were never detected. Twenty-seven dominating LAB strains were identified within the genera Enterococcus, Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Levilactobacillus, Lactococcus, Leuconostoc, Pediococcus and Streptococcus. This work showed that the wooden table used during the production of PDO Provola dei Nebrodi cheese is a safe system and a microbiologically active toolopenBusetta, Gabriele; Garofalo, Giuliana; Mangione, Guido; Botta, Luigi; Franciosi, Elena; Di Gerlando, Rosalia; Todaro, Massimo; Licitra, Giuseppe; Scatassa, Maria Luisa; Gaglio, Raimondo; Settanni, LucaBusetta, G.; Garofalo, G.; Mangione, G.; Botta, L.; Franciosi, E.; Di Gerlando, R.; Todaro, M.; Licitra, G.; Scatassa, M.L.; Gaglio, R.; Settanni, L