Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques

Background: Tomato brown rugose fruit virus (ToBRFV) is a highly infectious tobamovirus that causes severe disease in tomato (Solanum lycopersicum L.) crops. In Italy, the first ToBRFV outbreak occurred in 2018 in several provinces of the Sicily region. ToBRFV outbreak represents a serious threat for tomato crops in Italy and the Mediterranean Basin. Methods: Molecular and biological characterisation of the Sicilian ToBRFV ToB-SIC01/19 isolate was performed, and a sensitive and specific Real-time RT-PCR TaqMan minor groove binder probe method was developed to detect ToBRFV in infected plants and seeds. Moreover, four different sample preparation procedures (immunocapture, total RNA extraction, direct crude extract and leaf-disk crude extract) were evaluated. Results: The Sicilian isolate ToB-SIC01/19 (6,391 nt) showed a strong sequence identity with the isolates TBRFV-P12-3H and TBRFV-P12-3G from Germany, Tom1-Jo from Jordan and TBRFV-IL from Israel. The ToB-SIC01/19 isolate was successfully transmitted by mechanical inoculations in S. lycopersicum L. and Capsicum annuum L., but no transmission occurred in S. melongena L. The developed real-time RT-PCR, based on the use of a primer set designed on conserved sequences in the open reading frames3, enabled a reliable quantitative detection. This method allowed clear discrimination of ToBRFV from other viruses belonging to the genus Tobamovirus, minimising false-negative results. Using immunocapture and total RNA extraction procedures, the real-time RT-PCR and end-point RT-PCR gave the same comparable results. Using direct crude extracts and leaf-disk crude extracts, the end-point RT-PCR was unable to provide a reliable result. This developed highly specific and sensitive real-time RT-PCR assay will be a particularly valuable tool for early ToBRFV diagnosis, optimising procedures in terms of costs and time

The Relationship of Immune Status to the Efficacy of Immunotherapy in Preventing Tumour Recurrence in Mice

The immunotherapeutic value of tumour extracts or B.C.G. in preventing either the occurrence of primary tumours or the recurrence of tumours in surgically resected animals has been examined. A transplantable methylcholanthrene induced tumour in DBA/2J mice was used. Neither tumour extract nor chemically modified extract was effective in preventing tumour growth in immunized animals, even though the mice demonstrated measurable levels of cell mediated tumour immunity at the time of tumour challenge. The frequency of tumour recurrence after resection of small tumours (about 1·0 g) was significantly lowered by treatment of the mice with a combination of B.C.G. and either modified or unmodified tumour extract. The frequency of recurrence after resection of large tumours (about 2·5 g) was not affected by any form of immunotherapy although the survival time of treated animals was significantly prolonged. The immunological status of animals with small and large tumours was examined and it was shown that mice with 1·0 g tumours have unimpaired mitogen responsiveness and measurable tumour specific immunity, whereas mice bearing large tumours (2·5 g) have a markedly impaired immune system

Nature reappraisers, benefits for the environment: A model linking cognitive reappraisal, the “being away” dimension of restorativeness and eco-friendly behavior

In the last decades, an increasingly prominent role has been given to the motivational factors that can promote pro-environmental behavior. In this contribution, we focus on the role of the individual’s ability to shape the emotions originating from nature in engaging in pro-environmental behavior. In particular, we expect that an emotion regulation strategy as cognitive reappraisal should positively predict pro-environmental behavior, through enhanced perceived restorativeness attributed to the natural environment in terms of the experience of “being away.” One-hundred and fifteen visitors to an urban park (Parco Nord Milano) filled out a questionnaire including measures of cognitive reappraisal, the experience of “being away,” and pro-environmental behaviour while in the park. Results confirmed that cognitive reappraisal was positively and significantly related to pro-environmental behavior. Importantly, the indirect effect of cognitive reappraisal on pro-environmental behavior through the experience of “being away” was significant. Findings suggest the importance of implementing interventions aimed at promoting the habitual use of cognitive reappraisal to enhance the experience of “being away” and, thus, sustain pro-environmental behavior

Detection of Parietaria Mottle Virus by RT-qPCR: An Emerging Virus Native of Mediterranean Area That Undermine Tomato and Pepper Production in Southern Italy

Parietaria mottle virus (PMoV) is considered an emerging virus in many countries of the Mediterranean basin, especially on tomato and pepper crops. Symptoms on tomato leaves and fruits can be easily confused with those induced by cucumber mosaic virus (CMV) with necrogenic satellite RNA (CMV-satRNA), tomato spotted wilt virus (TSWV) or tomato mosaic virus (ToMV). Mixed infection of these viruses has been also reported in some tomato cultivars, with an increase in the complexity of the symptoms and severity of the disease. Although a specific serum and riboprobes have been produced, nowadays no sensitive diagnostic methods are available for the rapid PMoV detection. Here, we have developed a RT-qPCR assay with the aim to establish a more sensitive and specific method for PMoV detection. Specific primers and TaqMan probe were designed and in silico tested with all PMoV isolates available in GenBank. Moreover, this method was evaluated on tomato naturally infected samples from Sicily region (Italy). Results obtained showed that the RT-qPCR assay developed in this work is extremely sensitive, in fact, it is able to detect as few as 10 PMoV RNA copies in tomato total RNA; moreover, it will be a particularly valuable tool for early detection of PMoV. Furthermore, the analyzes on field samples show how this pathogen is increasingly present in tomato crops in the last years, helping to undermine the Italian horticultural sector

Public perceptions of forests across Italy: An exploratory national survey

In a context of progressive expansion of the Italian forest area, we present the results of a national survey exploring public perception of forests across different geographical scales in Italy. Perceptions of forests are assessed in rela-tion to popular beliefs on relevant environmental issues such as countering climate change, protecting biodiversity, and promoting social cohesion and environmental education. Participants (N = 1059) living in five different regions of Northern (Trentino-Alto Adige/Südtirol, Piemonte), Central (Lazio, Molise) and Southern Italy (Puglia), were recruited in the survey and completed a paper-and-pencil questionnaire. Survey questions regarded the estimated percentage of forest cover, the perceived importance of different environmental issues and of different material and non-material forest products, as well as partici-pants’ perceptions regarding connectedness to nature. Results revealed a gen-eralized tendency to overestimate the extension of forest surface area in the participants’ region, in Italy, and in the European Union. Results also showed high scores for participants’ perceived importance of environmental issues, such as climate change and biodiversity protection, and in their belief that forests could play a positive role in addressing these issues and providing im-portant outcomes and benefits for the quality of human life, such as health and well-being or social cohesion

Road Thermal Collector for Building Heating in South Europe: Numerical Modeling and Design of an Experimental Set-Up

The combination/integration of renewable energy and storage systems appears to have significant potential, achieving high-energy results with lower costs and emissions. One way to cover the thermal needs of a building is through solar energy and its seasonal storage in the ground. The SMARTEP project aims to create an experimental area that provides for the construction of a road solar thermal collector directly connected to a seasonal low-temperature geothermal storage with vertical boreholes. The storage can be connected to a ground-to-water heat pump for building acclimatization. This system will meet the requirements of visual impact and reduction of the occupied area. Nevertheless, several constraints related to the radiative properties of the surfaces and the lack of proper thermal insulation have to be addressed. The project includes the study of several configurations and suitable materials, the set-up of a dynamic simulation model and the construction of a small-scale road thermal collector. These phases allowed for an experimental area to be built. Thanks to careful investigation in the field, it will be possible to identify the characteristics and the best operation strategy to maximize the energy management of the whole system in the Mediterranean area

Are community gardening and horticultural interventions beneficial for psychosocial well-being? A meta-analysis

Recent literature has revealed the positive effect of gardening on human health; however, empirical evidence on the effects of gardening-based programs on psychosocial well-being is scant. This meta-analysis aims to examine the scientific literature on the effect of community gardening or horticultural interventions on a variety of outcomes related to psychosocial well-being, such as social cohesion, networking, social support, and trust. From 383 bibliographic records retrieved (from 1975 to 2019), seven studies with a total of 22 effect sizes were selected on the basis of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Meta-analytic findings on 11 comparisons indicate a positive and moderate effect of horticultural or gardening interventions on psychosocial well-being. Moderation analysis shows a greater effect size in individualistic than collectivistic cultures. A greater effect size was also observed in studies involving community gardening compared to horticultural intervention. Nevertheless, an effect of publication bias and study heterogeneity has been detected. Despite the presence of a large number of qualitative studies on the effect of horticulture/gardening on psychosocial well-being, quantitative studies are lacking. There is a strong need to advance into further high-quality studies on this research topic given that gardening has promising applied implications for human health, the community, and sustainable city management

Spread of tomato brown rugose fruit virus in sicily and evaluation of the spatiotemporal dispersion in experimental conditions

Tomato brown rugose fruit virus (ToBRFV) is an emerging pathogen that causes severe disease in tomato (Solanum lycopersicum L.) crops. The first ToBRFV outbreak in Italy occurred in 2018 in several Sicilian provinces, representing a serious threat for tomato production. In the present work, the spatiotemporal displacement of ToBRFV in Sicily was evaluated, analyzing a total of 590 lots of tomato seed, 982 lots of plantlets from nurseries and 100 commercial greenhouses. Furthermore, we investigated the ToBRFV spreading dynamic in a greenhouse under experimental conditions. Results showed several aspects related to ToBRFV dispersion in protected tomato crops. In detail, an important decrease of the ToBRFV-infected seed and plantlet lots was detected. Regarding the examined commercial greenhouses, ToBRFV still appears to be present in Sicily, although there has been a decrease during monitoring. In experimental conditions, it was demonstrated that the presence of few infected plants are sufficient to damage the entire crop in a short time, reaching almost 100% of infection