First report of pseudopithomyces mori causing leaf spot on mandarin in Italy

In July 2023, a leaf spot-like disease was observed on approximately 15% plants in a mandarin (Citrus reticulata) orchard in Calabria, Southern Italy. The symptomatic leaves displayed irregular, dark brown lesions surrounded by a yellow halo. Fifteen symptomatic leaves from five trees were collected and surface sterilized in 2% NaOCl. Lesion pieces (3 × 3 mm) were plated on potato dextrose agar (PDA) with antibiotics and incubated at 24 °C for 5 days. Three isolates morphologically identical were obtained from different plants, and purified as required. The colonies on PDA were cottony, first hyaline, then salmon pink (front) to orange (reverse). Conidia were ellipsoid, light brown, with 3–4 transverse septa, usually divided by a longitudinal septum, and 23.5 ± 5.5 × 13.69 ± 4.79 μm. The morphological features were consistent with those of Pseudopithomyces sp. (Ariyawansa et al. 2015). Identification was confirmed for the representative strain AP77 by sequencing the internal transcribed spacer (ITS) region and portion of translation elongation factor 1 alpha (TEF1) gene with primers ITS1/ITS4 and EF-983 F/EF-2218R, respectively (Jayasiri et al. 2019). Sequences were deposited in GenBank with accession numbers OR911598 (ITS) and OR921209 (TEF1). BLASTn analyses showed their 99% homology with Pseudopithomyces mori strain MFLUCC 18-1630 (GenBank No. MW183777 and MW063153, respectively). Phylogenetic analyses of the concatenated sequences confirmed the clustering with P. mori strain MFLUCC 18-1630 (Tennakoon et al. 2022). The pathogenicity was tested on 10 healthy citrus leaves, which were slightly injured with a sterile scalpel, inoculated by a drop of suspension (106 conidia/ml) or sterile water (control), and maintained at 24 °C in a humid chamber. Seven days after inoculation, lesions like those observed in the field occurred only on inoculated leaves. The same fungus was reisolated from infected leaves, fulfilling Koch’s postulates. To our knowledge, this is the first report of leaf spot on mandarin leaves caused by P. mori in Italy

Evaluation of a quality improvement intervention to reduce anastomotic leak following right colectomy (EAGLE): pragmatic, batched stepped-wedge, cluster-randomized trial in 64 countries

Background Anastomotic leak affects 8 per cent of patients after right colectomy with a 10-fold increased risk of postoperative death. The EAGLE study aimed to develop and test whether an international, standardized quality improvement intervention could reduce anastomotic leaks. Methods The internationally intended protocol, iteratively co-developed by a multistage Delphi process, comprised an online educational module introducing risk stratification, an intraoperative checklist, and harmonized surgical techniques. Clusters (hospital teams) were randomized to one of three arms with varied sequences of intervention/data collection by a derived stepped-wedge batch design (at least 18 hospital teams per batch). Patients were blinded to the study allocation. Low- and middle-income country enrolment was encouraged. The primary outcome (assessed by intention to treat) was anastomotic leak rate, and subgroup analyses by module completion (at least 80 per cent of surgeons, high engagement; less than 50 per cent, low engagement) were preplanned. Results A total 355 hospital teams registered, with 332 from 64 countries (39.2 per cent low and middle income) included in the final analysis. The online modules were completed by half of the surgeons (2143 of 4411). The primary analysis included 3039 of the 3268 patients recruited (206 patients had no anastomosis and 23 were lost to follow-up), with anastomotic leaks arising before and after the intervention in 10.1 and 9.6 per cent respectively (adjusted OR 0.87, 95 per cent c.i. 0.59 to 1.30; P = 0.498). The proportion of surgeons completing the educational modules was an influence: the leak rate decreased from 12.2 per cent (61 of 500) before intervention to 5.1 per cent (24 of 473) after intervention in high-engagement centres (adjusted OR 0.36, 0.20 to 0.64; P < 0.001), but this was not observed in low-engagement hospitals (8.3 per cent (59 of 714) and 13.8 per cent (61 of 443) respectively; adjusted OR 2.09, 1.31 to 3.31). Conclusion Completion of globally available digital training by engaged teams can alter anastomotic leak rates. Registration number: NCT04270721 (http://www.clinicaltrials.gov)