Development of Real-Time PCR for in wood-detection of Ceratocystis platani, the agent of canker stain of Platanus spp.

Ceratocystis platani is a quarantine fungal pathogen agent of canker stain, a destructive disease affecting Platanus. Despite its diagnosis being critical for disease control, there is still no effective diagnostic tool as all known mycological and biological detection assays are problematical. In this study we developed highly effective Real-Time PCR methods based on the use of an intercalating dye, EvaGreen, and on a Taqman probe. We designed primers and probe on the internal transcribed spacer (ITS) region of rDNA, and used them for the amplification and detection of a 95?bp C. platani amplicon. Inference of standard curves revealed that both Real-Time procedures have similar and high values of amplification efficiency when applied to a range of templates, e.g. genomic fungal DNA and DNA extracted from diseased wood. The methods were sensitive with a detection limit of 10?fg???l^?1? C. platani genomic DNA. They were specific as they did not yield any detection signal when applied to non-target fungal taxa colonizers of Platanus wood. Reliability was demonstrated through the positive detection of a collection of C. platani isolates and of wood samples collected from naturally infected trees. Robustness was positively verified through detection on artificially infected trees, which were tested at different times after death, up to 27?months. Generating a standard curve with a target-amplicon-containing plasmid enabled an absolute quantification and a comparison between the discoloured wood of resistant and susceptible genotypes. The importance of the methods for studies on pathogen epidemiology and host resistance is also discussed.     

Prevalence and risk factors of astrovirus infection in puppies from French breeding kennels

Aiming at determining the prevalence and the risk factors associated to astrovirus infection in puppy, fecal samples were collected in 316 puppies (age from 5 to 14 weeks of age) from 33 French breeding kennels. Data were registered for each puppy, including age, breed, gender, origin of the dog, and feces quality. The samples were tested by specific RT-PCR for the presence of canine astrovirus. Astroviruses were identified in 20.9% (66/316) of the puppies and in 42% (14/33) of the breeding kennels. Young puppies (i.e. <7 weeks of age) and puppies from large breeding kennels were more likely to be infected by the astrovirus. No association between the quality of feces and astrovirus infection could be determined in this survey.     

Molecular genotyping of Salmonella enterica Abortusovis by pulsed field gel electrophoresis

Genotyping of Salmonella strains is an important tool to discriminate among isolates and to improve epidemiological studies when an outbreak occurs. No phagetyping scheme is available for Salmonella enterica subsp. enterica serovar Abortusovis (SAO) and molecular methods previously used were not standardized and were time consuming. Among the DNA-based methods of genotyping, pulsed field gel electrophoresis (PFGE) is currently in use to subtype Salmonella isolates. In this study we evaluated the feasibility of genotyping of SAO by XbaI and BlnI restrictions. Separation of restricted fragments was performed by PFGE. To test the possibility to apply this methodology to epidemiological investigation, a collection of 38 SAO strains isolated in different regions of Italy were analyzed. Eighteen and 29 different PFGE profiles were defined for XbaI and BlnI digestions, respectively. The method demonstrated an adequate typing ability and an excellent discriminatory power. Results from this study show that PFGE may represent a powerful tool to discriminate within the SAO serovar, and provide useful information in support of traditional epidemiological investigations. In particular, this method could be used to identify the origin of infection during outbreaks within a single flock or in different herds.     

Digestive enzyme activity and nutrient digestibility in meagre (Argyrosomus regius) fed increasing levels of black soldier fly meal (Hermetia illucens)

The effect of partially replacing fishmeal (FM) by black soldier fly larvae meal (Hermetia meal—HM) in meagre (Argyrosomus regius) diet was evaluated for nutrient digestibility and digestive enzyme activity. For that purpose, triplicate groups of fish (18.0 ± 0.02 g) were fed during 48 days either a control diet (CTR), without HM, or one of three diets including 100, 200 and 300 g/kg of HM, replacing 17, 35 and 52% of FM, respectively. Apparent digestibility coefficients (ADCs) of dry matter, energy, protein, lysine, isoleucine, leucine, phenylalanine, alanine, glutamate, glycine and serine presented a decreasing response with increased HM inclusion. Chitin ADC was null, independent of dietary HM inclusion. Total alkaline protease activity increased while trypsin activity decreased with dietary HM inclusion. No intestinal chitinolytic activity was detected. Intestinal alkaline protease zymogram revealed nine bands with proteolytic activity against casein, with molecular weights ranging between 15 and 75 kDa. Anti‐protease activity in the intestine was not affected by dietary inclusion of HM compared to the CTR diet. Overall, it is concluded that replacement of up to 17% FM with HM (100 g/kg HM inclusion level) in meagre diets has no major adverse effects in diet digestibility and digestive enzyme activity.     

Residents and urban green spaces: The case of Bari

The total area of public green spaces in the city of Bari, Italy is more limited than in most other Italian cities (2.9 m²/inhabitant). This fact makes it an interesting subject for research into the general perception that the residents of the city have of green spaces and their behaviour patterns when using them.

A questionnaire (27 questions) was presented by telephone to a representative sample (n=351) of the population of Bari. The aims of the study were: (a) to gain insight in the perception of green spaces with particular reference to those green areas within the city itself; (b) to examine behaviour during visits to parks and gardens and means of transport used to reach them; (c) to investigate what kind of relationship should be set up between the local authority and the population regarding information and participation.

The results show that respondents perceive the green areas as a life quality enhancer in accordance with some previous studies carried out in Italy and in other countries. Citizens are moreover conscious of the limitations in quality and quantity of green areas in their own city, although this result needs to be more closely analysed in the future. Patterns in the use of public green areas proved to be strictly connected with age, sex, marital status, area of residence. Another conditioning factor was the system of mobility. The results moreover underline how citizens see the public and private green areas as single beneficial system.

The research also demonstrates the potentials of applying telephone surveys in studies concerning urban forests.     

Investigating the effects of afforestation on soil erosion and sediment mobilisation in two small catchments in Southern Italy

Annual soil losses in southern Italy can exceed 100–150 t ha^− 1 year^− 1. Where erosion on agricultural land is particularly severe, land use change and afforestation are frequently seen as the most appropriate means of reducing erosion risk. However, the overall effectiveness of afforestation in reducing soil erosion remains uncertain, due to the poor development of the forest cover in some areas, leading to significant areas with sparse tree cover, and the erosional impact of forest harvesting, which commonly involves clearcutting. The study reported here addresses this uncertainty and focuses on two small catchments (W2 and W3) located in Calabria, southern Italy, for which measurements of suspended sediment yield are available. Both the catchments originally supported a rangeland vegetation cover and they were planted with eucalyptus trees in 1968. Currently, only catchment W3 supports a continuous forest cover. In catchment W2 the forest cover is discontinuous and there is a significant area of the catchment (ca. 20%) where the tree cover is sparse and the vegetation cover is dominated by natural grasses. Two additional erosion plots were established within catchment W2 in 1991, in order to explore the effect of the density of the tree cover on soil erosion. Information on the sediment yields from the two catchments and the plots for 10 storm events that occurred during the period December 2005–December 2006 and associated information on the ¹³⁷Cs and excess ²¹⁰Pb of the sediment, have been used to investigate the effectiveness of afforestation in reducing sediment mobilisation and net soil loss from the catchments involved. The results demonstrate that the areas of greatest soil loss are associated with the slopes where the tree cover is discontinuous, and that forest harvesting by clearcutting causes significant short-term increases in sediment mobilisation and sediment yield. These findings, which are consistent with previous work undertaken within the same area, emphasize the importance of vegetation cover density in influencing rates of soil loss in the study catchments. The study also provided a useful demonstration of the potential for using measurements of the ¹³⁷Cs and ²¹⁰Pb_ex content of sediment, in combination with more traditional sediment monitoring, to investigate sediment sources and to compare the sediment dynamics of catchments subjected to different land management practices.     

Comparative topical anti-inflammatory activity of cannabinoids and cannabivarins

A selection of seven phytocannabinoids representative of the major structural types of classic cannabinoids and their corresponding cannabivarins was investigated for in vivo topical anti-inflammatory activity in the Croton oil mouse ear dermatitis assay. Differences in the terpenoid moiety were far more important for anti-inflammatory activity than those at the C-3 alkyl residue, suggesting the involvement not only of cannabinoid receptors, but also of other inflammatory end-points targeted by phytocannabinoids.     

Comparison of direct and indirect genetic methods for estimating seed and pollen dispersal in Fagus sylvatica and Fagus crenata

The comparison between estimates of historical gene flow, using variance in allelic frequencies, and estimates of contemporary gene flow, using parentage assignment, is expected to provide insights into ecological and evolutionary processes at work within and among populations. Genetic variation at microsatellite loci was used to quantify genetic structure in two wind pollinated, gravity and animal-dispersed tree species (Fagus sylvatica L. and Fagus crenata Blum.) and to derive historical estimates of gene flow. The gene dispersal distances estimated assuming effective population density to be 1/4 of the observed density were ∼77 m in European beech and ∼40 m in Japanese beech. Parentage analyses and a neighbourhood model approach were used to estimate contemporary patterns of seed and pollen dispersal. Our results suggest restricted seed dispersal abilities in both European beech (δ_s = 10.5 m) and Japanese beech (δ_s = 12.4 m), with an exponential shaped seed dispersal kernel. A non-negligible rate of seed immigration (m_s = 27%) was detected in European beech sites but not in Japanese beech site. Pollen dispersal within studied sites also appeared limited (δ_p = 41.63 m in European beech and δ_p = 79.4 m in Japanese beech), despite high rate of pollen immigration (m_p = 68% in European beech and m_p = 40% m in Japanese beech). Interestingly, contemporary and historical estimates of gene flow were within the same order of magnitude (a few tens of meters).