Use of Coniothyrium minitans transformed with the hygromycin B resistance gene to study survival and infection of Sclerotinia sclerotiorum sclerotia in soil

A Coniothyrium minitans strain (T3) co-transformed with the genes for β-glucuronidase (uidA) and hygromycin phosphotransferase (hph), the latter providing resistance to the antibiotic hygromycin B, was used to investigate the survival and infection of sclerotia of Sclerotinia sclerotiorum by C. minitans over time in four different soils. Infection of sclerotia was rapid in all cases, with the behaviour of transformant T3 and wild type parent A69 being similar. Differences were seen between the soils in the rate of infection of sclerotia by C. minitans and in their indigenous fungal populations. Amendment of agar with hygromycin B enabled the quantification of C. minitans in soil by dilution plating where there was a high background of other microorganisms. In Lincoln soil from New Zealand, which had a natural but low population of C. minitans the hygromycin B resistance marker allowed the umambiguous discrimination of the applied transformed isolate from the indigenous hygromycin B sensitive one. In this soil, although the indigenous C. minitans population was detected from sclerotia, none were recovered on the dilution plates, indicating the increased sensitivity of C. minitans detection from soil using sclerotial baiting. C. minitans was a very efficient parasite, being able to infect a large proportion of sclerotia within a relatively short time from an initially low soil population. The addition of hygromycin B to agar also allowed the detection of C. minitans from decaying sclerotia by inhibiting secondary fungal colonisers. This is the first report to show that fungi colonising sclerotia already infected by C. minitans mask the detection of C. minitans from sclerotia rather than displacing the original parasite

Ascospore release and survival in Sclerotinia sclerotiorum

The release and survival of ascospores of a UK Sclerotinia sclerotiorum isolate were studied. Apothecia placed in a spore clock apparatus with different lighting regimes at 15 °C released ascospores continuously with an increasing rate for the duration of experiments (72–84 h). Spore release was not confined to light or dark periods in alternating regimes and occurred in continuous dark or light. Ascospores were released in both saturated air (90–95% rh) and at 65–75% rh. High temperature and rh were detrimental to ascospore survival but spore viability was maintained for longer periods than previously reported. The significance of these results in relation to disease control is discussed

A New Species of \u3ci\u3eMyxidium\u3c/i\u3e (Myxosporea: Myxidiidae), from the Western Chorus Frog, \u3ci\u3ePseudacris triseriata triseriata\u3c/i\u3e, and Blanchard\u27s Cricket Frog, \u3ci\u3eAcris crepitans blanchardi\u3c/i\u3e (Hylidae), from Eastern Nebraska: Morphology, Phylogeny, and Critical Comments on Amphibian \u3ci\u3eMyxidium\u3c/i\u3e Taxonomy

During March 2001-April 2004, 164 adult anurans of 6 species (47 Rana blairi, 35 Rana catesbeiana, 31 Hyla chrysoscelis, 31 Pseudacris triseriata triseriata, 11 Bufo woodhousii, and 9 Acris crepitans blanchardi) from Pawnee Lake, Lancaster County, Nebraska, were surveyed for myxozoan parasites. Of these, 20 of 31 (65%) P. triseriata triseriata and 1 of 9 (11%) A. crepitans blanchardi were infected with a new species of Myxidium. Myxidium melleni n. sp. (Myxosporea) is described from the gallbladder of the western chorus frog, P. triseriata triseriata (Hylidae). This is the second species of Myxidium described from North American amphibians. Mature plasmodia are disc-shaped or elliptical 691 (400-1,375) × 499 (230-1,200) × 23 (16-35) μm, polysporic, producing many disporic pansporoblasts. The mature spores, 12.3 (12.0-13.5) × 7.6 (7.0-9.0) × 6.6 (6.0-8.0) μm, containing a single binucleated sporoplasm, are broadly elliptical, with 2-5 transverse grooves on each valve, and contain two equal polar capsules 5.2 (4.8-5.5) × 4.2 (3.8-4.5) μm positioned at opposite ends of the spore. Myxidium melleni n. sp. is morphologically consistent with other members of Myxidium. However, M. melleni n. sp. was phylogenetically distinct from other Myxidium species for which DNA sequences are available. Only with improved morphological analyses, accompanied by molecular data, and the deposit of type specimens, can the ambiguous nature of Myxidium be resolved. Guidelines for descriptions of new species of Myxidium are provided

Phylogeny of fish-infecting Calyptospora species (Apicomplexa: Eimeriorina)

There are numerous species of apicomplexans that infect poikilothermic vertebrates, such as fishes, and possess unique morphological features that provide insight into the evolution of this important phylum of parasites. Here, the relationship of the fish-infecting Calyptospora species to other coccidians was investigated based on DNA sequence analysis. Genetic data from the small subunit ribosomal DNA region of the genome were obtained for three of the five nominal species in the genus Calyptospora. Phylogenetic analyses supported a monophyletic lineage sister to a group composed of mostly Eimeria species. The monophyly of Calyptospora species supports the validity of the family Calyptosporidae, but the sister relationship to Eimeria species might also suggest the Eimeriidae be expanded to encompass Calyptospora. The validity of the family Calyptosporidae has been questioned because it is delineated from the Eimeriidae largely based on life cycle characteristics and sporocyst morphology. In general, Eimeria species have a homoxenous life cycle, whereas the type species of Calyptospora is heteroxenous. In the absence of experimental transmission studies, it may be difficult to demonstrate whether all Calyptospora species are heteroxenous. Other distinct morphological characteristics of Calyptospora such as an incomplete sporocyst suture, an apical opening for sporozoite release, a thin veil surrounding sporocysts supported by sporopodia, and a lack of Stieda and sub-Stieda bodies suggest there may be adequate features to delineate these taxa. Even without life cycle data for all species, the morphology and genetic data provide a means to reliably classify Calyptospora species. Placement in either the Calyptosporidae or Eimeriidae is discussed, along with issues relating to the phylogeny of the genus Goussia

Mortality and pulmonary complications in patients undergoing surgery with perioperative SARS-CoV-2 infection: an international cohort study

Background: The impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on postoperative recovery needs to be understood to inform clinical decision making during and after the COVID-19 pandemic. This study reports 30-day mortality and pulmonary complication rates in patients with perioperative SARS-CoV-2 infection. Methods: This international, multicentre, cohort study at 235 hospitals in 24 countries included all patients undergoing surgery who had SARS-CoV-2 infection confirmed within 7 days before or 30 days after surgery. The primary outcome measure was 30-day postoperative mortality and was assessed in all enrolled patients. The main secondary outcome measure was pulmonary complications, defined as pneumonia, acute respiratory distress syndrome, or unexpected postoperative ventilation. Findings: This analysis includes 1128 patients who had surgery between Jan 1 and March 31, 2020, of whom 835 (74·0%) had emergency surgery and 280 (24·8%) had elective surgery. SARS-CoV-2 infection was confirmed preoperatively in 294 (26·1%) patients. 30-day mortality was 23·8% (268 of 1128). Pulmonary complications occurred in 577 (51·2%) of 1128 patients; 30-day mortality in these patients was 38·0% (219 of 577), accounting for 81·7% (219 of 268) of all deaths. In adjusted analyses, 30-day mortality was associated with male sex (odds ratio 1·75 [95% CI 1·28–2·40], p\textless0·0001), age 70 years or older versus younger than 70 years (2·30 [1·65–3·22], p\textless0·0001), American Society of Anesthesiologists grades 3–5 versus grades 1–2 (2·35 [1·57–3·53], p\textless0·0001), malignant versus benign or obstetric diagnosis (1·55 [1·01–2·39], p=0·046), emergency versus elective surgery (1·67 [1·06–2·63], p=0·026), and major versus minor surgery (1·52 [1·01–2·31], p=0·047). Interpretation: Postoperative pulmonary complications occur in half of patients with perioperative SARS-CoV-2 infection and are associated with high mortality. Thresholds for surgery during the COVID-19 pandemic should be higher than during normal practice, particularly in men aged 70 years and older. Consideration should be given for postponing non-urgent procedures and promoting non-operative treatment to delay or avoid the need for surgery. Funding: National Institute for Health Research (NIHR), Association of Coloproctology of Great Britain and Ireland, Bowel and Cancer Research, Bowel Disease Research Foundation, Association of Upper Gastrointestinal Surgeons, British Association of Surgical Oncology, British Gynaecological Cancer Society, European Society of Coloproctology, NIHR Academy, Sarcoma UK, Vascular Society for Great Britain and Ireland, and Yorkshire Cancer Research

Notes for genera: basal clades of Fungi (including Aphelidiomycota, Basidiobolomycota, Blastocladiomycota, Calcarisporiellomycota, Caulochytriomycota, Chytridiomycota, Entomophthoromycota, Glomeromycota, Kickxellomycota, Monoblepharomycota, Mortierellomycota, Mucoromycota, Neocallimastigomycota, Olpidiomycota, Rozellomycota and Zoopagomycota)

Compared to the higher fungi (Dikarya), taxonomic and evolutionary studies on the basal clades of fungi are fewer in number. Thus, the generic boundaries and higher ranks in the basal clades of fungi are poorly known. Recent DNA based taxonomic studies have provided reliable and accurate information. It is therefore necessary to compile all available information since basal clades genera lack updated checklists or outlines. Recently, Tedersoo et al. (MycoKeys 13:1--20, 2016) accepted Aphelidiomycota and Rozellomycota in Fungal clade. Thus, we regard both these phyla as members in Kingdom Fungi. We accept 16 phyla in basal clades viz. Aphelidiomycota, Basidiobolomycota, Blastocladiomycota, Calcarisporiellomycota, Caulochytriomycota, Chytridiomycota, Entomophthoromycota, Glomeromycota, Kickxellomycota, Monoblepharomycota, Mortierellomycota, Mucoromycota, Neocallimastigomycota, Olpidiomycota, Rozellomycota and Zoopagomycota. Thus, 611 genera in 153 families, 43 orders and 18 classes are provided with details of classification, synonyms, life modes, distribution, recent literature and genomic data. Moreover, Catenariaceae Couch is proposed to be conserved, Cladochytriales Mozl.-Standr. is emended and the family Nephridiophagaceae is introduced

Use of fungal antagonists for biological control of damping-off and scierotial pathogens

<div>Lecture given by Doctor John M. Whipps, AFRC Institute of Horticultural Research, Littlehampton at the Waite Campus, University of Adelaide, 21.9.1993</div><div><br></div><p>This lecture is part of the Campus Seminars and Distinguished Lecturer Series, Waite Campus, University of Adelaide, 1991 – 1997. </p> <a href="https://figshare.com/projects/Campus_Seminars_and_Distinguished_Lecturer_Series_Waite_Campus_University_of_Adelaide/17756">https://figshare.com/projects/Campus_Seminars_and_Distinguished_Lecturer_Series_Waite_Campus_University_of_Adelaide/17756</a

Beneficial microorganism survival on seed, roots and in rhizosphere soil following application to seed during drum priming

Priming is a technique used to improve seedling establishment of direct-seeded crops such as onion and carrot, resulting in a quick and uniform emergence. This work investigated the application of four selected beneficial microorganisms (Pseudomonas chlororaphis MA342, Pseudomonas fluorescens CHA0, Clonostachys rosea IK726d11 and Trichoderma harzianum T22) to onion and carrot seed during drum priming, and their subsequent survival and establishment in the rhizosphere once the seed was planted. Different application rates of fungi (7log(10) cfu g(-1) dry seed) and bacteria (6log(10) cfu g(-1) dry seed) were required on onion to achieve the end target of 5log10 cfu g(-1) dry seed, whereas a lower rate (5log(10) cfu g(-1) dry seed for both bacteria and fungi) was successful on carrot. Microorganism-treated seed was planted in soil in the glasshouse and root and rhizosphere soil samples were taken at 2, 4 and 8 weeks post-planting. All seed-applied microorganisms were recovered throughout the experiment, although differences in the survival patterns were seen. The bacterial isolates declined in number over time, with P. fluorescens CHA0 showing better overall survival than P. chlororaphis MA342, particularly on the roots and in the rhizosphere soil of carrot. In contrast to the bacteria, the fungal isolate C rosea IK726d11 showed good survival on both onion and carrot, and increased significantly in number throughout the 8-week period. Trichoderma harzianum T22 remained relatively constant in number throughout the experiment, but showed better survival on carrot than onion roots. Similar results were found in three different soil-types. (C) 2007 Elsevier Inc. All rights reserved