Assessment of bioelectrical phase angle as a predictor of nutritional status in patients with Crohn's disease: A cross sectional study

Background & aims: The assessment of body composition (BC) can be used to identify malnutrition in patients with Crohn's disease (CD). The aim of this study was to evaluate the nutritional status of CD patients by assessing BC, phase angle (PhA) and muscle strength. Differences in disease duration and medications were also considered. Methods: Consecutive adult CD patients aged 18–65 years were enrolled in this cross-sectional study. Disease activity was clinically defined by the Crohn's Disease Activity Index (CDAI) in the active and quiescent phases. All participants underwent anthropometry, BC and handgrip-strength (HGS) measurements; additionally, blood samples were taken. Data from CD patients were also compared with age-, sex- and BMI-matched healthy people. Results: A total of 140 CD patients with a mean age of 38.8 ± 13.9 years and a mean body weight of 64.9 ± 12 kg were recruited and compared to controls. The findings showed that all nutritional parameters, especially PhA and HGS, were lower in CD patients than in controls, and these parameters were substantially impaired as disease activity increased. Active CD patients had a lower body weight and fat mass than both the quiescent and control groups. PhA was negatively correlated with age (r = −0.362; p = 0.000) and CDAI (r = −0.135; p = 0.001) but was positively associated with fat free mass (FFM) (r = 0.443; p = 0.000) and HGS (r = 0.539; p = 0.000). Similarly, serum protein markers were lower in the active CD group than in the quiescent group (p < 0.05). Disease duration and medications did not significantly affect nutritional status. Conclusions: BIA-derived PhA is a valid indicator of nutritional status in CD patients, and its values decreased with increasing disease activity. Additionally, small alterations in BC, such as low FFM, and reduced HGS values can be considered markers of nutritional deficiency. Therefore, the assessment of BC should be recommended in clinical practice for screening and monitoring the nutritional status of CD patients

Detection and identification of phytoplasmas in two vineyards located in a restricted geographic area in Italy

A survey to verify the presence of grapevine yellows was carried out in a restricted area of the Marche region (central east Italy) in a small geographic area. In two locations six grapevine varieties were examined and phytoplasmas belonging to four ribosomal groups were detected by PCR/RFLP analyses. The surveyed varieties were Passerina, Pecorino, Merlot, Sangiovese, Fedit and Montepulciano. The different phytoplasmas detected were present in a scattered manner in all the varieties, but only Passerina, Pecorino and Sangiovese where shown to be infected by “bois noir” (16SrXII-A) phytoplasmas. Considering the landscape shape and the age of the vineyards the source of BN infection and of the other phytoplasmas is very likely in the surrounding environment or vineyards

Isolation of diverse phytoplasmas from symptomatic grapevine samples

Phytoplasmas from different ribosomal groups were isolated from grapevine samples in which “flavescence dorée” or “bois noir” phytoplasmas had been identified. The results indicate that the developed medium is not phytoplasma specific and supports the growth of phytoplasmas which cannot be detected by other molecular methods and are very likely present in a very low titre in the endobiome of the plants. The applied method discriminates the presence of bacterial contamination already in the liquid phase, and the colony purification technique allows excluding the contamination

Molecular variability and host distribution of ‘candidatus phytoplasma solani’ strains from different geographic origins

The knowledge of phytoplasma genetic variability is a tool to study their epidemiology and to implement an effective monitoring and management of their associated diseases. ‘Candidatus Phytoplasma solani’ is associated with “bois noir” disease in grapevines, and yellowing and decline symptoms in many plant species, causing serious damages during the epidemic outbreaks. The epidemiology of the diseases associated with this phytoplasma is complex and related to numerous factors, such as interactions of the host plant and insect vectors and spreading through infected plant propagation material. The genetic variability of ‘Ca. P. solani’ strains in different host species and in different geographic areas during the last two decades was studied by RFLP analyses coupled with sequencing on vmp1, stamp, and tuf genes. A total of 119 strains were examined, 25 molecular variants were identified, and the variability of the studied genes was linked to both geographic distribution and year of infection. The crucial question in ‘Ca. P. solani’ epidemiology is to trace back the epidemic cycle of the infections. This study presents some relevant features about differential strain distribution useful for disease monitoring and forecasting, illustrating and comparing the phytoplasma molecular variants identified in various regions, host species, and time periods

Eggplant Little Leaf-Associated Phytoplasma Detection in Seedlings under Insect-Proof Conditions

Eggplant, or brinjal, is one of the most consumed and important tropical solanaceous veg- etable crops grown worldwide. Little leaf is a disease associated with the presence of phytoplasmas especially widespread in brinjal in India. To clarify the epidemiology of this disease, a verifica- tion of its transmission through seeds to seedlings and their progeny derived from symptomatic mother plants was performed. Brinjal seeds field-collected in the Dharwad district of Karnataka State, India, were sowed in a greenhouse under insect-proof conditions. DNA was extracted from seedlings and their progeny and from symptomatic plant samples collected in the field. The first- and second-generation seedlings obtained *under these conditions were tested at various time points after germination by amplification of the 16S rRNA gene of phytoplasmas. The amplicons obtained were subjected to restriction fragment length polymorphism (RFLP) analysis and sequencing for the identification of detected phytoplasmas. Ribosomal groups 16SrI, 16SrII, 16SrIII, 16SrV, 16SrVI, and 16SrXII were identified. Moreover, a number of fruits produced from the first-generation seedlings showed precocious seed germination, and the young seedlings resulted as phytoplasma-positive. The seed transmission of phytoplasmas in eggplants for two subsequent generations highlights the risk of additional sources of infection of the disease represented by asymptomatic and infected seedlings in the presence of insect vectors. The seed transmission could explain the continuous presence of epidemic outbreaks of phytoplasmas in brinjal cultivations in several cultivation areas

Survey for ‘Candidatus Liberibacter’ and ‘Candidatus Phytoplasma’ in Citrus in Chile

The considerable economic losses in citrus associated with ‘Candidatus Liberibacter’ and ‘Candidatus Phytoplasma’ presence have alerted all producing regions of the world. In Chile, none of these bacteria have been reported in citrus species. During the years 2017 and 2019, 258 samples presenting symptoms similar to those associated with the presence of these bacteria were examined. No detection of ‘Ca. Liberibacter’ associated with “huanglongbing” disease was obtained in the tested samples; therefore, this quarantine pest is maintained as absent in Chile. However, 14 plants resulted positive for phytoplasmas enclosed in subgroups 16SrV-A (12 plants) and 16SrXIII-F (2 plants). Although they have been found in other plant species, this is the first report of these phy-toplasmas in citrus worldwide

Small RNA Profiling of Aster Yellows Phytoplasma-Infected Catharanthus roseus Plants Showing Different Symptoms

Micropropagated Catharantus roseus plants infected with 'Candidatus Phytoplasma asteris' showed virescence symptoms, witches' broom symptoms, or became asymptomatic after their planting in pots. Nine plants were grouped into three categories according to these symptoms, which were then employed for investigation. The phytoplasma concentration, as determined by qPCR, correlated well with the severity of symptoms. To reveal the changes in the small RNA profiles in these plants, small RNA high-throughput sequencing (HTS) was carried out. The bioinformatics comparison of the micro (mi) RNA and small interfering (si) RNA profiles of the symptomatic and asymptomatic plants showed changes, which could be correlated to some of the observed symptoms. These results complement previous studies on phytoplasmas and serve as a starting point for small RNA-omic studies in phytoplasma research

Molecular identification of ‘Candidatus Phytoplasma palmicola’ associated with coconut lethal yellowing in Equatorial Guinea

During the past two decades, a high mortality of coconut palms was observed in the coastal areas of Equatorial Guinea. Reportedly, the palm population has been reduced by 60%-70%, and coconut production has decreased accordingly. To identify the cause of the mortality, a survey was carried out in April 2021 in various localities of the coconut belt. Molecular analyses carried out on 16S rRNA and secA genes detected phytoplasma presence in the majority of the samples. Sequencing and BLAST search of the 16S rRNA gene sequences showed >99% identity of the detected phytoplasmas to 'Candidatus Phytoplasma palmicola'. The RFLP analyses of 16S ribosomal gene using Tru1I and TaqI enzymes led to assign these phytoplasmas to subgroup 16SrXXII-A. In all samples that tested positive, including one from a hybrid coconut palm and two from oil palm the same phytoplasma was identified. The phylogenetic analyses of 16S rRNA and secA genes confirmed respectively 99.98%-100% and 97.94%-100% identity to 'Ca. P. palmicola'. RFLP analyses using MboII enzyme on the secA gene amplicon differentiated the phytoplasma found in Equatorial Guinea from those present in Ghana and Ivory Coast. The Equatorial Guinean phytoplasma strain resulted to be identical to the strains from Mozambique, confirming the presence of a geographic differentiation among phytoplasma strains in the coastal areas of Western and Central Africa. The identified phytoplasma is different from the 'Ca. P. palmicola' strains found in Ghana and Ivory Coast and represents the first identification a 16SrXXII-A strain in Equatorial Guinea and in Central Africa. Strict monitoring and surveillance procedures for early detection of the pathogen are strongly recommended to reduce its impact and further spread in the country and permit the recovery of coconut plantations

Induction of resistance and enhancing agronomic performance in grapevines under greenhouse and in open fields by applications of plasma activated water

The exposure of water to a cold atmospheric pressure plasma (CAP) enables the production of plasma activated water (PAW), having high content of reactive species, whose applications were tested on grapevine plants, both in greenhouse and in vineyard conditions. Two different CAPs were used for PAW production, evaluating their effectiveness as a possible mean to control plant diseases. Grapevines infected with yellows associated with the presence of phytoplasmas were treated evaluating qualitative and quantitative yield parameters, phytoplasma presence, and gene expression. The results show the capability of PAW to enhance plant defence mechanisms and, as demonstrated in the field trials, confirmed its ability to improve the health status of the treated plants. Quantitative (q)RT-PCR analyses allowed to determine the transcription level of genes involved in the plant defence response (phenylalanine ammonia-lyase, pal) and in the plant phytoalexin metabolism of PAW-treated materials. The number of symptomatic grapevine plants in vineyards was significantly reduced by the treatments. Transcriptional and post-transcriptional molecular analyses highlighted the PAW ability to enhance the expression of genes encoding the main enzymes involved in the phytoalexin biosynthetic pathway (flavonoids and stilbenes). The PAW ability to enhance some of the plant defence mechanisms also improving the health status of the treated plants was therefore experimentally demonstrated. After three years of trials the overall results demonstrated the possible use of PAW to reduce the disease severity, induce plant resistance both in open field and greenhouse, improving plant healthy status and grapevine yield production

Transcriptional profiling of phytoplasma infected plants treated with plasma activated water (PAW).

Background. Phytoplasmas are insect-transmitted plant pathogenic prokaryotes, associated with severe diseases in agronomic important crops. Management of these diseases has mainly focused on insect vector chemical control and on infected plant rouging. There is therefore a strong need for effective and friendly control strategies for phytoplasma-associated diseases and the possibility to use plasma activated water (PAW) as sustainable and effective method to them was therefore evaluated. PAW is produced by treating distilled water with atmospheric pressure plasmas, inducing the production of reactive oxygen and nitrogen species (RONS) and pH reduction. PAW has good potential for bacterial decontamination, degradation of organic compounds and was shown to positively affect plant growth. Methods. Sterile deionized water (SDW) was exposed to a nanosecond pulsed dielectric barrier discharge, operating in ambient air for 10 min treatment with a peak voltage of 19 kV and a pulse repetition frequency of 1 kHz, which induced production of nitrates, nitrites and peroxides, and a pH decrease. Phytoplasma infected and healthy periwinkles micropropagated shoots were exposed to PAW for about 25 minutes and gene expression studies were then performed. The theses used were: shoots treated with PAW, Fosetyl aluminum (as positive control) and SDW (as negative control), with an exposition of about 25 minutes. Nine shoots for each thesis were then collected at 6 different times after treatment and stored at -80\ub0C. Quantitative RT-PCR analyses were carried out to determine the expression level of genes involved in the plant defense response. Parallel experiments were carried out treating grapevine plants in vineyards previously tested for the phytoplasma presence. Treatments were performed for three years injecting into the plant vascular tissues 10-20 ml of PAW or sterile distilled water (as control) on each selected plant for a total of 60 plants (40 with phytoplasmas and 20 without phytoplasmas). Results. Overexpression of selected genes involved in the phytoalexin metabolism was detected in the periwinkles micropropagated shoots treated with PAW in comparison with the shoots treated with Fosetyl-Al and distilled water. In the field trials, in a relevant number of cases, the PAW-treated symptomatic plants showed reduction of symptoms, while the SDW-treated and untreated plants did not show symptom reduction. No phytotoxicity was observed in the PAW treated grapevine and periwinkle plants. Conclusion. The results obtained showed the capability of PAW to enhance plant defence mechanisms and, as demonstrated in the field trials, confirmed its ability to improve the health status of the treated plant