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This mixed-methods study aims to investigate the bias in peer feedback. Thirty-two English as an additional language learners gave each other anonymous feedback on their texts. Half of the participants received feedback from their teacher disguised as peer feedback, while the other half received actual peer feedback. Data were collected through drafts of two essays, feedback, and a questionnaire. Results indicate that although participants reported trusting teacher feedback more, the quantitative uptake of feedback was similar regardless of the source. Data analysis suggests that the teacher's and peers' social representation plays a more significant role in uptake than the feedback itself.

BACKGROUND: Fungal plant pathogens have dynamic genomes that allow them to rapidly adapt to adverse conditions and overcome host resistance. One way by which this dynamic genome plasticity is expressed is through effector gene loss, which enables plant pathogens to overcome recognition by cognate resistance genes in the host. However, the exact nature of these loses remains elusive in many fungi. This includes the tomato pathogen Cladosporium fulvum, which is the first fungal plant pathogen from which avirulence (Avr) genes were ever cloned and in which loss of Avr genes is often reported as a means of overcoming recognition by cognate tomato Cf resistance genes. A recent near-complete reference genome assembly of C. fulvum isolate Race 5 revealed a compartmentalized genome architecture and the presence of an accessory chromosome, thereby creating a basis for studying genome plasticity in fungal plant pathogens and its impact on avirulence genes. RESULTS: Here, we obtained near-complete genome assemblies of four additional C. fulvum isolates. The genome assemblies had similar sizes (66.96 to 67.78 Mb), number of predicted genes (14,895 to 14,981), and estimated completeness (98.8 to 98.9%). Comparative analysis that included the genome of isolate Race 5 revealed high levels of synteny and colinearity, which extended to the density and distribution of repetitive elements and of repeat-induced point (RIP) mutations across homologous chromosomes. Nonetheless, structural variations, likely mediated by transposable elements and effecting the deletion of the avirulence genes Avr4E, Avr5, and Avr9, were also identified. The isolates further shared a core set of 13 chromosomes, but two accessory chromosomes were identified as well. Accessory chromosomes were significantly smaller in size, and one carried pseudogenized copies of two effector genes. Whole-genome alignments further revealed genomic islands of near-zero nucleotide diversity interspersed with islands of high nucleotide diversity that co-localized with repeat-rich regions. These regions were likely generated by RIP, which generally asymmetrically affected the genome of C. fulvum. CONCLUSIONS: Our results reveal new evolutionary aspects of the C. fulvum genome and provide new insights on the importance of genomic structural variations in overcoming host resistance in fungal plant pathogens.

Erysiphe necator is an obligate fungal pathogen that causes grape powdery mildew, globally the most important disease on grapevines. Previous attempts to obtain a quality genome assembly for this pathogen were hindered by its high repetitive DNA content. Here, chromatin conformation capture (Hi-C) with long-read PacBio sequencing was combined to obtain a chromosome-scale assembly and a high-quality annotation for E. necator isolate EnFRAME01. The resulting 81.1 Mb genome assembly is 98% complete and consists of 34 scaffolds, 11 of which represent complete chromosomes. All chromosomes contain large centromeric-like regions and lack synteny to the 11 chromosomes of the cereal PM pathogen Blumeria graminis. Further analysis of their composition showed that repeats and transposable elements (TEs) occupy 62.7% of their content. TEs were almost evenly interspersed outside centromeric and telomeric regions and massively overlapped with regions of annotated genes, suggesting that they could have a significant functional impact. Abundant gene duplicates were observed as well, particularly in genes encoding candidate secreted effector proteins. Moreover, younger in age gene duplicates exhibited more relaxed selection pressure and were more likely to be located physically close in the genome than older duplicates. A total of 122 genes with copy number variations among six isolates of E. necator were also identified and were enriched in genes that were duplicated in EnFRAME01, indicating they may reflect an adaptive variation. Taken together, our study illuminates higher-order genomic architectural features of E. necator and provides a valuable resource for studying genomic structural variations in this pathogen. IMPORTANCE Grape powdery mildew caused by the ascomycete fungus Erysiphe necator is economically the most important and recurrent disease in vineyards across the world. The obligate biotrophic nature of E. necator hinders the use of typical genetic methods to elucidate its pathogenicity and adaptation to adverse conditions, and thus comparative genomics has been a major method to study its genome biology. However, the current reference genome of E. necator isolate C-strain is highly fragmented with many non-coding regions left unassembled. This incompleteness prohibits in-depth comparative genomic analyses and the study of genomic structural variations (SVs) that are known to affect several aspects of microbial life, including fitness, virulence, and host adaptation. By obtaining a chromosome-scale genome assembly and a high-quality gene annotation for E. necator, we reveal the organization of its chromosomal content, unearth previously unknown features of its biology, and provide a reference for studying genomic SVs in this pathogen.

Studies have been widely carried out on drying techniques and equipment. The behavior of clays subjected to drying stages requires further research. This study aims to evaluate the behavior of the clay ceramic body during the different stages of the drying cycle and match the characteristics of the raw materials with the final quality of the pieces. In this work, the behavior of three different clays in terms of drying performance was studied. The clays were selected, and their chemical, mineralogical, and particle size characteristics were measured. The mixture design (DoE) developed 10 formulations and was processed through vacuum extrusion. The samples were subjected to forced drying cycles of 180 min by varying the temperature from 30 to 90 °C and air speed from 1.5 to 4.0 m/s. At the end of the cycle, the retraction was a determining factor for the crack probability indicator, where formulations that obtained ∼10% retraction in the dry zone showed losses >25%. On the other hand, it is could be stated that claystone assists the drying process of the ceramic piece, minimizing losses. The drying sensitivity coefficient (k-factor) presented values that reproduce the number of losses during drying, proving to be a valid tool to relate clay properties, drying conditions, and losses in this process. In this case, drying process losses >25% were observed when the k-factor was from 1.6.

Fulvia fulva and Dothistroma septosporum are closely related apoplastic pathogens with similar lifestyles but different hosts: F. fulva is a pathogen of tomato, whilst D. septosporum is a pathogen of pine trees. In 2012, the first genome sequences of these pathogens were published, with F. fulva and D. septosporum having highly fragmented and near-complete assemblies, respectively. Since then, significant advances have been made in unravelling their genome architectures. For instance, the genome of F. fulva has now been assembled into 14 chromosomes, 13 of which have synteny with the 14 chromosomes of D. septosporum, suggesting these pathogens are even more closely related than originally thought. Considerable advances have also been made in the identification and functional characterization of virulence factors (e.g., effector proteins and secondary metabolites) from these pathogens, thereby providing new insights into how they promote host colonization or activate plant defence responses. For example, it has now been established that effector proteins from both F. fulva and D. septosporum interact with cell-surface immune receptors and co-receptors to activate the plant immune system. Progress has also been made in understanding how F. fulva and D. septosporum have evolved with their host plants, whilst intensive research into pandemics of Dothistroma needle blight in the Northern Hemisphere has shed light on the origins, migration, and genetic diversity of the global D. septosporum population. In this review, we specifically summarize advances made in our understanding of the F. fulva-tomato and D. septosporum-pine pathosystems over the last 10 years.

Objective: The treatment of Hemophilia A and B has significantly evolved. Study design: we conducted a retrospective, observational study among patients with HA and HB managed in a Pediatric Hematology and Oncology Unit for almost 22 years for a total of 34 patients. Results: Until 2009, more than 80 % of patients were treated only on demand, while in 2022,100 % of patients with moderate and severe disease were receiving continuous prophylaxis. The median age at the start of prophylaxis passed from 10,9 to 1,65 years. In 2022, only one patient was on Standard-half-life-rFVIII, 7 patients were on Extended Half-Life (EHL)-rFVIII/rFIX and 7 patients were on Emicizumab; 1 patient was on Concizumab. The mean ABR passed from 5,7 in the period 2001–2009 to 0,8 in the period 2019–2022. We performed 106 ultrasonographic exams in 19 patients to evaluate the HEAD-US score: a percentage of 45,3 % of these exams revealed a score of 0 and the HEAD-US score resulted inferior in patients who started prophylaxis at an age below the median age at the initiation of prophylaxis. The score showed a tendency to correlate with the number of previous hemarthrosis. In our cohort, the overall prevalence of inhibitors is 0,09 among all patients, and 0,14 in severe HA/HB. 4 patients needed a Central Venous Line (CVL), and 3 patients developed at least one severe complication. Before 2017, we observed 4 cases of life-threatening bleeding in children less than 2 years old not on prophylaxis. Results: our real-life experience reflects the changes in hemophilia management in the last two decades. In the future, other drugs are expected.

Abstract Studies have proven that the use of waste in the production of clay ceramics is common for achieving the technical performance that is necessary for the application of these by-products. However, it is essential to analyze the feasibility of using these materials in terms of productivity and economic gains, in line with sustainability. The objective of this study is to determine the cost-benefit ratio of using rice husk ash in the production of clay ceramics. The methodological process consisted of a comparative analysis on an industrial scale between the formulation with up to 15% by volume of rice husk ash and the standard formulation previously used by the company. The economic aspects included the costs/consumption of raw materials, transport, and energy. The productivity throughout the process and technical characteristics of the final product were also evaluated. The costs of raw materials showed a financial gain of 7.0%, whereas the productivity analysis revealed that the greatest economic gain was in energy consumption during drying (13%).

Review: Resenha: LI, M. Researching and Teaching Second Language Writing in the Digital Age. Palgrave Macmillan: Cham, Switzerland, 2021. Disponível em: https://doi.org/10.1007/978-3-030-87710-1_4

Summary: Intra-tumoral phenotypic heterogeneity promotes tumor relapse and therapeutic resistance and remains an unsolved clinical challenge. Decoding the interconnections among different biological axes of plasticity is crucial to understand the molecular origins of phenotypic heterogeneity. Here, we use multi-modal transcriptomic data—bulk, single-cell, and spatial transcriptomics—from breast cancer cell lines and primary tumor samples, to identify associations between epithelial-mesenchymal transition (EMT) and luminal-basal plasticity—two key processes that enable heterogeneity. We show that luminal breast cancer strongly associates with an epithelial cell state, but basal breast cancer is associated with hybrid epithelial/mesenchymal phenotype(s) and higher phenotypic heterogeneity. Mathematical modeling of core underlying gene regulatory networks representative of the crosstalk between the luminal-basal and epithelial-mesenchymal axes elucidate mechanistic underpinnings of the observed associations from transcriptomic data. Our systems-based approach integrating multi-modal data analysis with mechanism-based modeling offers a predictive framework to characterize intra-tumor heterogeneity and identify interventions to restrict it.

Abstract Background Fungal plant pathogens have dynamic genomes that allow them to rapidly adapt to adverse conditions and overcome host resistance. One way by which this dynamic genome plasticity is expressed is through effector gene loss, which enables plant pathogens to overcome recognition by cognate resistance genes in the host. However, the exact nature of these loses remains elusive in many fungi. This includes the tomato pathogen Cladosporium fulvum, which is the first fungal plant pathogen from which avirulence (Avr) genes were ever cloned and in which loss of Avr genes is often reported as a means of overcoming recognition by cognate tomato Cf resistance genes. A recent near-complete reference genome assembly of C. fulvum isolate Race 5 revealed a compartmentalized genome architecture and the presence of an accessory chromosome, thereby creating a basis for studying genome plasticity in fungal plant pathogens and its impact on avirulence genes. Results Here, we obtained near-complete genome assemblies of four additional C. fulvum isolates. The genome assemblies had similar sizes (66.96 to 67.78 Mb), number of predicted genes (14,895 to 14,981), and estimated completeness (98.8 to 98.9%). Comparative analysis that included the genome of isolate Race 5 revealed high levels of synteny and colinearity, which extended to the density and distribution of repetitive elements and of repeat-induced point (RIP) mutations across homologous chromosomes. Nonetheless, structural variations, likely mediated by transposable elements and effecting the deletion of the avirulence genes Avr4E, Avr5, and Avr9, were also identified. The isolates further shared a core set of 13 chromosomes, but two accessory chromosomes were identified as well. Accessory chromosomes were significantly smaller in size, and one carried pseudogenized copies of two effector genes. Whole-genome alignments further revealed genomic islands of near-zero nucleotide diversity interspersed with islands of high nucleotide diversity that co-localized with repeat-rich regions. These regions were likely generated by RIP, which generally asymmetrically affected the genome of C. fulvum. Conclusions Our results reveal new evolutionary aspects of the C. fulvum genome and provide new insights on the importance of genomic structural variations in overcoming host resistance in fungal plant pathogens.

This mixed-methods study aims to investigate the bias in peer feedback. Thirty-two English as an additional language learners gave each other anonymous feedback on their texts. Half of the participants received feedback from their teacher disguised as peer feedback, while the other half received actual peer feedback. Data were collected through drafts of two essays, feedback, and a questionnaire. Results indicate that although participants reported trusting teacher feedback more, the quantitative uptake of feedback was similar regardless of the source. Data analysis suggests that the teacher’s and peers’ social representation plays a more significant role in uptake than the feedback itself.

Cladosporium fulvum is a fungal pathogen that causes leaf mould of tomato. The reference genome of this pathogen was released in 2012 but its high repetitive DNA content prevented a contiguous assembly and further prohibited the analysis of its genome architecture. In this study, we combined third generation sequencing technology with the Hi-C chromatin conformation capture technique, to produce a high-quality and near complete genome assembly and gene annotation of a Race 5 isolate of C. fulvum. The resulting genome assembly contained 67.17 Mb organized into 14 chromosomes (Chr1-to-Chr14), all of which were assembled telomere-to-telomere. The smallest of the chromosomes, Chr14, is only 460 kb in size and contains 25 genes that all encode hypothetical proteins. Notably, PCR assays revealed that Chr14 was absent in 19 out of 24 isolates of a world-wide collection of C. fulvum, indicating that Chr14 is dispensable. Thus, C. fulvum is currently the second species of Capnodiales shown to harbour dispensable chromosomes. The genome of C. fulvum Race 5 is 49.7 % repetitive and contains 14 690 predicted genes with an estimated completeness of 98.9%, currently one of the highest among the Capnodiales. Genome structure analysis revealed a compartmentalized architecture composed of gene-dense and repeat-poor regions interspersed with gene-sparse and repeat-rich regions. Nearly 39.2 % of the C. fulvum Race 5 genome is affected by Repeat-Induced Point (RIP) mutations and evidence of RIP leakage toward non-repetitive regions was observed in all chromosomes, indicating the RIP plays an important role in the evolution of this pathogen. Finally, 345 genes encoding candidate effectors were identified in C. fulvum Race 5, with a significant enrichment of their location in gene-sparse regions, in accordance with the 'two-speed genome' model of evolution. Overall, the new reference genome of C. fulvum presents several notable features and is a valuable resource for studies in plant pathogens.

Powdery mildews comprise a large group of economically important phytopathogenic fungi. However, limited information exists on their mitochondrial genomes. Here, we assembled and compared the mitochondrial genomes of the powdery mildew pathogens Blumeria graminis f. sp. tritici, Erysiphe pisi, and Golovinomyces cichoracearum. Included in the comparative analysis was also the mitochondrial genome of Erysiphe necator that was previously analysed. The mitochondrial genomes of the four Erysiphales exhibit a similar gene content and organization but a large variation in size, with sizes ranging from 109800 bp in B. graminis f. sp. tritici to 332165 bp in G. cichoracearum, which is the largest mitochondrial genome of a fungal pathogen reported to date. Further comparative analysis revealed an unusual bimodal GC distribution in the mitochondrial genomes of B. graminis f. sp. tritici and G. cichoracearum that was not previously observed in fungi. The cytochrome b (cob) genes of E. necator, E. pisi, and G. cichoracearum were also exceptionally rich in introns, which in turn harboured rare open reading frames encoding reverse transcriptases that were likely acquired horizontally. Golovinomyces cichoracearum had also the longest cob gene (45 kb) among 703 fungal cob genes analysed. Collectively, these results provide novel insights into the organization of mitochondrial genomes of powdery mildew pathogens and represent valuable resources for population genetics and evolutionary studies.

Mineral extraction of coal in the carboniferous region of southern Santa Catarina (Brazil) plays a significant role in the regional economy. However, this activity has severe environmental impacts, with approximately 65% of the extracted mineral being discarded as a rejected material (deposited in landfills). The identification of the technological potential of these materials, based on the geological aspects of the extraction site and the beneficiation operations applied to obtain coal, provides the opportunity to add value to different residual fractions that can be reused. Thus, waste valorization, the main objective of this work, has recently become a strategy for the application of these minerals in the production of clay ceramics using a systematic approach named CPQvA, which means “classification, potentiality, quantity/viability, and applicability”. The use of these materials as secondary mineral sources can avoid the deposition of these materials in industrial landfills and help to reduce the pressure on landfills, which receive an average of four million tons of material annually. In this study, the residual fraction, part of the tailing from coal beneficiation, known as coal fines, was evaluated for formulation valorization in clay ceramics. This residual fraction was classified as non-hazardous (class II-A, non-inert). X-ray fluorescence spectrometry, X-ray diffraction, and thermogravimetric analysis were performed to characterize the moisture content, particle-size distribution, and coal content to be used in the development of different formulations using the residual fraction of coal tailings (ranging from 0 to 40%) that are of technological interest to the sector. Processing parameters, such as firing at different temperatures (750, 800, 850, and 900 °C), were also correlated with these formulations. The results were compared with those of a reference ceramic formulation from the local productive arrangement of Morro da Fumaça (Arranjo Produtivo Local Cerâmica Vermelha de Morro da Fumaça). The various relationships between the materials were characterized in terms of their thermal shrinkage, water absorption, and mechanical resistance. Leaching and solubilization environmental tests revealed that both the industrial standard formulation and formulations with the application of the residual fraction were classified as non-hazardous materials. Thus, the method of using a mining residual fraction in the formulation of clay ceramics proved to be beneficial for the circular economy in the regional mineral sector through productive and environmental gains; the primary mineral resource and energy consumptions and the impacts related to waste generation were reduced. The results of this study can be applied to similar situations in other parts of the world.

Although repetition is at the core of many different approaches to language learning, either implicitly or not, using this pedagogic practice in the additional language classroom is still negatively perceived by some teachers (Bygate and Samuda, 2005). For contemporary research, on the other hand, the use of repetition is not incongruous with communicative additional language teaching approaches that bring the use of tasks to the forefront. The use of immediate repeated tasks can benefit learners because it allows the possibility of repeating slightly altered tasks in a meaningful way. Bearing this in mind, this paper describes three immediate repetition tasks that focus on the speaking skill aiming at developing both fluency and accuracy. All are inherently suited for the additional language classroom and can be easily adapted to better suit specific contexts.

This article aims to analyze whether formal instruction influences Brazilian speakers' perception of the English high back vowels contrast. There have been a few L2 pieces of research that focused on the instruction of specific vowel contrasts. Previous studies indicate that a single L1 category seems to be a source of difficulty to L2 vowel discrimination. However, some of these investigations did not focus on the role of instruction to such discrimination. The participants of the present study were 17 Brazilian speakers of Portuguese as L1, beginning learners of English, divided into experimental and control groups. The study included a perception pretest, a pronunciation instruction class, taught only to the experimental group, and a perception posttest. Results showed that experimental and control groups obtained similar results. Based on that, some factors were pointed to possibly explain this outcome, such as the duration of the pronunciation instruction, the possibility of participants learning with the pretest itself, the duration of the data collection, the participants' possible assimilation of the target contrast into a single category, and the interference of the mid central vowel /?/ used as a distractor in the data collection. On the other hand, a qualitative analysis revealed that all participants in the experimental group found the pronunciation instruction helpful. Such findings seem to agree entirely or in part with other similar studies' results.

Powdery mildews are notorious fungal plant pathogens but only limited information exists on their genomes. Here we present the mitochondrial genome of the grape powdery mildew fungus Erysiphe necator and a high-quality mitochondrial gene annotation generated through cloning and Sanger sequencing of full-length cDNA clones. The E. necator mitochondrial genome consists of a circular DNA sequence of 188,577 bp that harbors a core set of 14 protein-coding genes that are typically present in fungal mitochondrial genomes, along with genes encoding the small and large ribosomal subunits, a ribosomal protein S3, and 25 mitochondrial-encoded transfer RNAs (mt-tRNAs). Interestingly, it also exhibits a distinct gene organization with atypical bicistronic-like expression of the nad4L/nad5 and atp6/nad3 gene pairs, and contains a large number of 70 introns, making it one of the richest in introns mitochondrial genomes among fungi. Sixty-four intronic ORFs were also found, most of which encoded homing endonucleases of the LAGLIDADG or GIY-YIG families. Further comparative analysis of five E. necator isolates revealed 203 polymorphic sites, but only five were located within exons of the core mitochondrial genes. These results provide insights into the organization of mitochondrial genomes of powdery mildews and represent valuable resources for population genetic and evolutionary studies.

We prove spectral stability results for the $ curl curl $ operator subject to electric boundary conditions on a cavity upon boundary perturbations. The cavities are assumed to be sufficiently smooth but we impose weak restrictions on the strength of the perturbations. The methods are of variational type and are based on two main ingredients: the construction of suitable Piola-type transformations between domains and the proof of uniform Gaffney inequalities obtained by means of uniform a priori $ H^2 $-estimates for the Poisson problem of the Dirichlet Laplacian. The uniform a priori estimates are proved by using the results of V. Maz'ya and T. Shaposhnikova based on Sobolev multipliers. Connections to boundary homogenization problems are also indicated.

Using Swain's (1985) output hypothesis as a basis, this article investigated the effect an immediate repeated oral task had on the performance of participants. Two groups of beginner learners of English as an additional language in Brazil performed a decision-making oral task in groups. Drawing from Lynch and MacLean's (2001) carousel task, the learners changed groups and repeated the oral task in three successive cycles. A qualitative analysis of two learners' oral production is discussed in terms of accuracy. Furthermore, their perception on the immediate repeated task is also examined.

ABSTRACT The objective of this study was to evaluate whether the addition of different levels of capsaicin in the diet of female turkeys has beneficial effects on growth performance and the antioxidant and oxidant status in serum and meat. A total of 150 female turkeys were distributed in a completely randomized design with three treatments with five replicates per treatment and ten birds per experimental unit. Treatments were identified as control (without additive); T400 – basal diet containing 400 mg/kg of pepper extract per kg of feed; and T800 – basal diet supplemented with 800 mg/kg of pepper extract per kg of feed. Growth performance was measured on days 1 and 20 of the experiment. Blood samples were collected at 20 days of the experiment for analysis of the oxidant and antioxidant status, and at 20 days, five birds were euthanized per treatment for the analysis of the oxidant and antioxidant status of the meat. The turkeys fed diet supplemented with capsaicin had lower feed intake and better feed:gain ratio. Turkeys in the T800 treatment showed a reduction in serum levels of reactive oxygen species (ROS) and thiobarbituric acid, and this same effect was observed for ROS in their breast meat. Capsaicin supplementation improves feed conversion and reduces feed intake without altering weight gain in female turkeys. Furthermore, the addition of 800 g/ton reduces lipid peroxidation and protein oxidation in the serum and reduces protein oxidation in broiler turkey meat.

ABSTRACT The objective of this study was to evaluate whether the addition of the enzymes phytase and xylanase, isolated or associated, in the liquid form after feed pelletization could improve energy utilization and digestibility of calcium and phosphorus by broiler chickens. Three experiments were performed using 120 birds each, divided into five treatments with eight replicates per group (n = 3), identified as: experiment 1 (xylanase: control, 1000 IU, 1500 IU, 2000 IU, 2500 IU), experiment 2 (phytase: control, 500 FTU, 1000 FTU, 1500 FTU, 2000 FTU), experiment 3 (xylanase + phytase: control, 3000 IU + 500 FTU, 3000 IU + 1000 FTU, 3000 IU + 1500 FTU, 3000 IU + 2000 FTU). Samples for digestibility tests were collected at 14 to 21 days of age. Therefore, the inclusion of liquid phytase and liquid phytase + xylanase after pelletization in broiler diets has become a relevant way to reduce the inclusion of inorganic P, which can reduce the cost of feed and P excretion in the environment. Furthermore, it is an interesting strategy to avoid enzyme denaturation in the pelleting process.

Infectious keratitis is a sight-threatening condition that is usually an ocular emergency. The visual outcome depends on prompt and accurate clinical management as well as geographic and epidemiological awareness. We conducted a retrospective observational study to define the epidemiological and laboratory profile, as well as the clinical course of bacterial keratitis in a tertiary hospital in São Paulo over 21 years. Information about age, sex, predisposing factors, topical and surgical treatment, visual acuity, ulcers’ classification, bacterioscopy, culture, and antibiotic sensitivity tests were collected. This study included 160 patients. The mean age was 65.1 ± 18.4 years and risk factors were identified in 83.1 % of the patients. Empirical topical fortified cephalosporin with an aminoglycoside or fourth-generation fluoroquinolone was curative for 66.2 % of the cases. The mean treatment duration was 22.5 ± 9 days. The mean variation of visual acuity was -0.25 logMAR, p < 0.001. Culture revealed 64 % of Gram-positive bacteria. All Gram-positive bacteria were sensitive to cephalothin, vancomycin, and quinolones. All Gram-negative bacteria were sensitive to gentamicin, tobramycin, amikacin, and ciprofloxacin. These findings reinforce the importance of prompt empirical treatment of severe corneal ulcers with a fortified cephalosporin and aminoglycoside or a fourth-generation fluoroquinolone as there are equally effective. Collected data was insufficient to evaluate resistance of ocular infections over time in this population.

ABSTRACT Erysiphe necator is an obligate fungal pathogen that causes grape powdery mildew, globally the most important disease on grapevines. Previous attempts to obtain a quality genome assembly for this pathogen were hindered by its high repetitive DNA content. Here, chromatin conformation capture (Hi-C) with long-read PacBio sequencing was combined to obtain a chromosome-scale assembly and a high-quality annotation for E. necator isolate EnFRAME01. The resulting 81.1 Mb genome assembly is 98% complete and consists of 34 scaffolds, 11 of which represent complete chromosomes. All chromosomes contain large centromeric-like regions and lack synteny to the 11 chromosomes of the cereal PM pathogen Blumeria graminis. Further analysis of their composition showed that repeats and transposable elements (TEs) occupy 62.7% of their content. TEs were almost evenly interspersed outside centromeric and telomeric regions and massively overlapped with regions of annotated genes, suggesting that they could have a significant functional impact. Abundant gene duplicates were observed as well, particularly in genes encoding candidate secreted effector proteins. Moreover, younger in age gene duplicates exhibited more relaxed selection pressure and were more likely to be located physically close in the genome than older duplicates. A total of 122 genes with copy number variations among six isolates of E. necator were also identified and were enriched in genes that were duplicated in EnFRAME01, indicating they may reflect an adaptive variation. Taken together, our study illuminates higher-order genomic architectural features of E. necator and provides a valuable resource for studying genomic structural variations in this pathogen. IMPORTANCE Grape powdery mildew caused by the ascomycete fungus Erysiphe necator is economically the most important and recurrent disease in vineyards across the world. The obligate biotrophic nature of E. necator hinders the use of typical genetic methods to elucidate its pathogenicity and adaptation to adverse conditions, and thus comparative genomics has been a major method to study its genome biology. However, the current reference genome of E. necator isolate C-strain is highly fragmented with many non-coding regions left unassembled. This incompleteness prohibits in-depth comparative genomic analyses and the study of genomic structural variations (SVs) that are known to affect several aspects of microbial life, including fitness, virulence, and host adaptation. By obtaining a chromosome-scale genome assembly and a high-quality gene annotation for E. necator, we reveal the organization of its chromosomal content, unearth previously unknown features of its biology, and provide a reference for studying genomic SVs in this pathogen.

Worldwide, Ascochyta blight is caused by a complex of host-specific fungal pathogens, including Ascochyta pisi, Didymella pinodes, and Didymella pinodella. The application of foliar fungicides is often necessary for disease management, but a better understanding of pathogen prevalence, aggressiveness, and fungicide sensitivity is needed to optimize control. Leaf and stem samples were obtained from 56 field pea production fields in 14 counties in North Dakota from 2017 to 2020 and isolates were collected from lesions characteristic of Ascochyta blight. Based on fungal characteristics and sequencing the ITS1-5.8S-ITS2 region, 73% of isolates were confirmed to be D. pinodes (n = 177) and 27% were A. pisi (n = 65). Across pathogens, aggressiveness was similar among some isolates in greenhouse assays. The in vitro pyraclostrobin sensitivity of all D. pinodes isolates collected from 2017 to 2020 was lower than that of the three baseline isolates. Sensitivity of 91% of A. pisi isolates collected in 2019 and 2020 was lower than the sensitivity of two known sensitive isolates. Resistance factors (Rf) from mean EC50 values of pyraclostrobin baseline/known sensitive isolates to isolates collected from 2017 to 2020 ranged from 2 to 1,429 for D. pinodes and 1 to 209 for A. pisi. In vitro prothioconazole sensitivity of 91% of D. pinodes isolates collected from 2017 to 2020 was lower than the sensitivity of the baseline isolates and 98% of A. pisi isolates collected from 2019 to 2020 was lower than the sensitivity of the known sensitive isolates. Prothioconazole Rf ranged from 1 to 338 for D. pinodes and 1 to 127 for A. pisi. Based on in vitro results, 92% of D. pinodes and 98% of A. pisi isolates collected displayed reduced-sensitivity/resistance to both fungicides when compared to baseline/known sensitive isolates. Disease control under greenhouse conditions of both pathogens provided by both fungicides was significantly lower in isolates determined to be reduced-sensitive or resistant in in vitro assays when compared to sensitive. Results reported here reinforce growers desperate need of alternative fungicides and/or management tools to fight Ascochyta blight in North Dakota and neighboring regions.