Widespread distribution of kiwifruit bacterial canker caused by the European Pseudomonas syringae pv. actinidiae genotype in the main production areas of Portugal

In Portugal, in 2010, after the first recorded occurrence of Pseudomonas syringae pv. actinidiae (Psa) in a few kiwifruit orchards, and following subsequent records of suspected symptoms at the beginning of 2011, an extensive survey concerning Psa was carried out. One hundred and sixty-two samples (trunks, twigs, leaves, flowers, buds) presenting symptoms were collected from the main Portuguese kiwifruit areas and submitted to phyto-bacteriological analyses to verify possible relationships between the symptoms/damage observed and the presence of Psa in kiwifruit orchards. The results confirmed the widespread presence of the pathogen in the main production areas of Portugal, where different Actinidia spp. cultivars are grown. Genetic investigations revealed that all the Psa strains detected belong to the European genotype. Preliminary considerations concerning pathogen dissemination are made

Genetic Diversity of Pseudomonas syringae pv. actinidiae Strains from Different Geographic Regions in China

Pseudomonas syringae pv. actinidiae causes kiwifruit bacterial canker, with severe infection of the kiwifruit plant resulting in heavy economic losses. Little is known regarding the biodiversity and genetic variation of populations of P. syringae pv. actinidiae in China. A collection of 269 strains of P. syringae pv. actinidiae was identified from 300 isolates obtained from eight sampling sites in five provinces in China. The profiles of 50 strains of P. syringae pv. actinidiae and one strain of P. syringae pv. actinidifoliorum were characterized by Rep-, insertion sequences 50, and randomly amplified polymorphic DNA polymerase chain reaction (PCR). Discriminant analysis of principal coordinates, principal component analysis, and hierarchical cluster analysis were used to analyze the combined fingerprints of the different PCR assays. The results revealed that all isolates belonged to the Psa3 group, that strains of P. syringae pv. actinidiae from China have broad genetic variability that was related to source geographic region, and that Chinese strains can be readily differentiated from strains from France but are very similar to those from Italy. Multilocus sequence typing of 24 representative isolates using the concatenated sequences of five housekeeping genes (cts, gapA, gyrB, pfk, and rpoD) demonstrated that strain Jzhy2 from China formed an independent clade compared with the other biovars, which possessed the hopH1 effector gene but lacked the hopA1 effector gene. A constellation analysis based on the presence or absence of the four loci coding for phytotoxins and a cluster analysis based on the 11 effector genes showed that strains from China formed two distinct clades. All of the strains, including K3 isolated in 1997 from Jeju, Korea, lacked the cfl gene coding for coronatine. In contrast, the tox-argK gene cluster coding for phaseolotoxin was detected in K3 and in the biovar 1 strains (K3, Kw30, and Psa92), and produced a false-positive amplicon for the hopAM1-like gene in this study. To date, only one biovar (biovar 3) is represented by the strains of P. syringae pv. actinidiae from China, despite China being the center of origin for kiwifruit

Genetic diversity and population structure of Pseudomonas savastanoi, an endemic pathogen of the Mediterranean area, revealed up to strain level by the MLVA assay

© 2020, Società Italiana di Patologia Vegetale (S.I.Pa.V.). Pseudomonas savastanoi is a bacterial species included in the Pseudomonas syringae complex. It is further sub-typed in pathovars which are the causal agents of a group of diseases of woody plants, such as the “knot disease” on olive and oleander and the bacterial canker on ash. Given its long-established presence in the Mediterranean area, the pathogen causing the afore-mentioned diseases can be considered endemic. Here, an MLVA approach was developed to assess the genetic relationships among and within those pathovars, with a specific focus on P. savastanoi pv. savastanoi. By analyzing the genome of the P.savastanoi pv. savastanoi strain NCPPB 3335 (accession n° CP008742), 14 Tandem Repeat (TR) loci were identified and the corresponding primers were designed and used for the amplification of genomic DNAs from 84 strains belonging to Pseudomonas savastanoi pathovars. Data were analyzed using different approaches, such as hierarchical clustering, STRUCTURE, and k-means clustering with DAPC to evaluate the effectiveness of the assay in describing pathovars and population structure of the pathogen. Results reveal a very complex articulation of genetic relationships, as expected from a long-time evolving pathogen, while providing the possibility to discriminate the pathovars between each other. At intra-pathovar level, the MLVA assay clusters isolates mainly according to their hosts and geographic origin. This resulted particularly useful in the identification and tracking of P. savastanoi populations at local level.11n

The Italian inter-laboratory study on the detection of Pseudomonas syringae pv. actinide

A severe form of bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae (Psa), has been detected in all the main areas of cultivation of kiwifruit (Actinidia deliciosa and A. chinensis). Since 2010 several research groups have been assessing methods and procedures to detect and identify Psa, both from symptomatic and symptomless host material. In 2011, a study to compare Psa diagnostic methods was performed with reference to Psa strains and related pathovars, and with plant extracts or DNA obtained from healthy and naturally infected leaves, pollen or wood. The study revealed the strengths and the weaknesses of the assessed methods. The procedure included screening tests for Psa detection and for identification of Psa colonies. The methods assessed were bacterial isolation on generic and semi-selective media, PCR analysis (single, duplex and rep-PCR assay, the latter for identification only). The results highlighted the best performance of semi-selective with respect the generic media; the usefulness of the direct-PCR as screening tests for Psa detection; and the greater specificity of duplex-PCR and sensitivity of simple-PCR. The use of semi-selective medium for isolation and of two PCR-based methods - in parallel - for Psa detection are suggested. Both rep-PCR and duplex-PCR, were found to be specific, and are recommended as an identification test for this pathogen

Impact of COVID-19 Lockdown, during the Two Waves, on Drug Use and Emergency Department Access in People with Epilepsy: An Interrupted Time-Series Analysis

BACKGROUND: In 2020, during the COVID-19 pandemic, Italy implemented two national lockdowns aimed at reducing virus transmission. We assessed whether these lockdowns affected anti-seizure medication (ASM) use and epilepsy-related access to emergency departments (ED) in the general population. METHODS: We performed a population-based study using the healthcare administrative database of Tuscany. We defined the weekly time series of prevalence and incidence of ASM, along with the incidence of epilepsy-related ED access from 1 January 2018 to 27 December 2020 in the general population. An interrupted time-series analysis was used to assess the effect of lockdowns on the observed outcomes. RESULTS: Compared to pre-lockdown, we observed a relevant reduction of ASM incidence (0.65; 95% Confidence Intervals: 0.59-0.72) and ED access (0.72; 0.64-0.82), and a slight decrease of ASM prevalence (0.95; 0.94-0.96). During the post-lockdown the ASM incidence reported higher values compared to pre-lockdown, whereas ASM prevalence and ED access remained lower. Results also indicate a lower impact of the second lockdown for both ASM prevalence (0.97; 0.96-0.98) and incidence (0.89; 0.80-0.99). CONCLUSION: The lockdowns implemented during the COVID-19 outbreaks significantly affected ASM use and epilepsy-related ED access. The potential consequences of these phenomenon are still unknown, although an increased incidence of epilepsy-related symptoms after the first lockdown has been observed. These findings emphasize the need of ensuring continuous care of epileptic patients in stressful conditions such as the COVID-19 pandemic

Antidepressants Drug Use during COVID-19 Waves in the Tuscan General Population: An Interrupted Time-Series Analysis

In Italy, during the COVID-19 waves two lockdowns were implemented to prevent virus diffusion in the general population. Data on antidepressant (AD) use in these periods are still scarce. This study aimed at exploring the impact of COVID-19 lockdowns on prevalence and incidence of antidepressant drug use in the general population. A population-based study using the healthcare administrative database of Tuscany was performed. We selected a dynamic cohort of subjects with at least one ADs dispensing from 1 January 2018 to 27 December 2020. The weekly prevalence and incidence of drug use were estimated across different segments: pre-lockdown (1 January 2018-8 March 2020), first lockdown (9 March 2020-15 June 2020), post-first lockdown (16 June 2020-15 November 2020) and second lockdown (16 November 2020-27 December 2020). An interrupted time-series analysis was used to assess the effect of lockdowns on the observed outcomes. Compared to the pre-lockdown we observed an abrupt reduction of ADs incidence (Incidence-Ratio: 0.82; 95% Confidence-Intervals: 0.74-0.91) and a slight weekly decrease of prevalence (Prevalence-Ratio: 0.997; 0.996-0.999). During the post-first lockdown AD use increased, with higher incidence- and similar prevalence values compared with those expected in the absence of the outbreak. This pandemic has impacted AD drug use in the general population with potential rebound effects during the period between waves. This calls for future studies aimed at exploring the mid-long term effects of this phenomenon

An Innovative Superconducting Magnetic Trap for Probing β-decay in Plasmas

The main aim of Plasmas for Astrophysics Nuclear Decays Observation and Radiation for Archaeometry (PANDORA) project is to build a compact and flexible magnetic plasma trap where plasma reaches a densityne∼ 1011–1013 cm−3, and a temperature, in units ofkT,kTe∼ 0.1–30 keV in order to measure, for the first time, nuclearβ-decay rates in stellar-like conditions. One of the most important aspects of an ECR Ion Source (ECRIS) is its magnetic system. In this paper, the numerical design of the PANDORA magnetic system is presented and validated by using the commercial simulators OPERA and CST Studio Suite, showing an excellent agreement between each other in terms of axial and radial field profiles. In conjunction to the magnetic system design, the overall injection system, including the microwave lines for plasma heating and the isotopes injection schemes with a focus on the developments of the oven for solid elements, has been conceived and will be discussed

Design study of a HPGe detector array for β-decay investigation in laboratory ECR plasmas

In the frame of the PANDORA project, a new experimental approach aims at measuring in-plasma β-decay rate as a function of thermodynamical conditions of the environment, namely a laboratory magnetized plasma able to mimic some stellar-like conditions. The decay rates (expected to change dramatically as a function of the ionization state) will be measured as a function of the charge state distribution of the in-plasma ions. The new experimental approach aims at correlating the plasma environment and the decay rate. This can be performed by simultaneously identifying and discriminating—through an innovative multi-diagnostic system working synergically with a γ-ray detection system —the photons emitted by the plasma and γ-rays emitted after the isotope β-decay. In this study, the numerical simulations supporting the design of the γ-ray detector array, including a statistical significance study to check the feasibility of measuring the in-plasma decay rates, are presented. Geant4 simulations focused on the design of the array of γ-ray detectors and the evaluation of total efficiency depending on the detector type and the optimal displacement of detectors around the trap (including collimation systems and shielding). The simulation results showed that, due to technical limitations in the number of apertures that can be created in the magnetic trap, the best compromise is to use 14 HPGe (70% of relative efficiency) detectors surrounding the magnetic trap. The HPGe detectors were chosen for their excellent energy resolution (0.2% @ 1 MeV), since the harsh environment (the background is represented by the intense plasma self-emission) strongly affects the signal-to-background ratio. Once determined the total photopeak efficiency (0.1–0.2%), the sensitivity of the PANDORA experiment was checked in a "virtual experimental run," by exploring the measurability of isotope decay rates for the first three physical cases of PANDORA: 176Lu, 134Cs and 94Nb. The preliminary results demonstrated the feasibility of the measurement in terms of the signal-to-background ratio and significance that it is possible to reach. The results indicated that experimental run durations could take from several days to 3 months, depending on the isotope under investigation, thus shedding new light on the role of weak interactions in stellar nucleosynthesis

Experimental and numerical investigation of magneto-plasma optical properties toward measurements of opacity relevant for compact binary objects

Electromagnetic transients known as kilonovae (KN), are among the photonic messengers released in the post-merger phase of compact binary objects, for example, binary neutron stars, and they have been recently observed as the electromagnetic counterpart of related gravitational-wave (GW) events. Detection of the KN signal plays a fundamental role in the multi-messenger astronomy entering in a sophisticated GW-detecting network. The KN light curve also delivers precious information on the composition and dynamics of the neutron-rich post-merger plasma ejecta (relying on r-process nucleosynthesis yields). In this sense, studying KN becomes of great relevance for nuclear astrophysics. Because of the highly heterogeneous composition, plasma opacity has a great impact both on radiative transport and spectroscopic observation of KN. Theoretical models attempting in encoding the opacity of this system often fail, due to the complexity of blending plethora of both light- and heavy-r nuclei transition lines, requesting for more complete atomic database. Trapped magneto-plasmas conceived in PANDORA could answer to these requests, allowing experimental in-laboratory measurements of optical properties and opacities, at plasma electron densities and temperatures resembling early-stage plasma ejecta’s conditions, contributing to shed light on r-process metallic species abundance at the blue-KN diffusion time. A numerical study has been recently performed, supporting the choice of first physics cases to be investigated and the design of the experimental setup. In this article, we report on the feasibility of metallic plasmas on the basis of the results from the systematic numerical survey on optical spectra computed under non-local thermodynamic equilibrium (NLTE) for several light-r nuclei. Results show the great impact of the NLTE regime of laboratory magneto-plasmas on the gray opacity contribution contrasted with those under the astrophysical LTE assumption. A first experimental attempt of reproducing ejecta plasma conditions has been performed on the operative Flexible Plasma Trap (FPT) at the INFN-LNS and here presented, together with first plasma characterization of density and temperature, via non-invasive optical emission spectroscopy (OES). The measured plasma parameters have supported numerical simulations to explore optical properties of NLTE gaseous and metallic plasmas, in view of the near-future plasma opacity measurements through spectroscopic techniques. The novel work so far performed on these under-dense and low-temperature magneto-plasmas, opens the route for the first-time to future in-laboratory plasma opacity measurements of metallic plasma species relevant for KN light curve studies

Pseudomonas syringae pv. actinidiae (PSA) Isolates from Recent Bacterial Canker of Kiwifruit Outbreaks Belong to the Same Genetic Lineage

Intercontinental spread of emerging plant diseases is one of the most serious threats to world agriculture. One emerging disease is bacterial canker of kiwi fruit (Actinidia deliciosa and A. chinensis) caused by Pseudomonas syringae pv. actinidiae (PSA). The disease first occurred in China and Japan in the 1980s and in Korea and Italy in the 1990s. A more severe form of the disease broke out in Italy in 2008 and in additional countries in 2010 and 2011 threatening the viability of the global kiwi fruit industry. To start investigating the source and routes of international transmission of PSA, genomes of strains from China (the country of origin of the genus Actinidia), Japan, Korea, Italy and Portugal have been sequenced. Strains from China, Italy, and Portugal have been found to belong to the same clonal lineage with only 6 single nucleotide polymorphisms (SNPs) in 3,453,192 bp and one genomic island distinguishing the Chinese strains from the European strains. Not more than two SNPs distinguish each of the Italian and Portuguese strains from each other. The Japanese and Korean strains belong to a separate genetic lineage as previously reported. Analysis of additional European isolates and of New Zealand isolates exploiting genome-derived markers showed that these strains belong to the same lineage as the Italian and Chinese strains. Interestingly, the analyzed New Zealand strains are identical to European strains at the tested SNP loci but test positive for the genomic island present in the sequenced Chinese strains and negative for the genomic island present in the European strains. Results are interpreted in regard to the possible direction of movement of the pathogen between countries and suggest a possible Chinese origin of the European and New Zealand outbreaks