Identification and Confirmation of Virulence Factor Production from , a Causal Agent of Root Rot in Pulses.

is an aggressive pathogen of pulse crops and a causal agent in root rot disease that negatively impacts Canadian agriculture. This study reports the results of a targeted metabolomics-based profiling of secondary metabolism in an 18-strain panel of cultured axenically in multiple media conditions, in addition to an in planta infection assay involving four strains inoculated on two pea cultivars. Multiple secondary metabolites with known roles as virulence factors were detected which have not been previously associated with , including fungal decalin-containing diterpenoid pyrones (FDDPs), fusaoctaxins, sambutoxin and fusahexin, in addition to confirmation of previously reported secondary metabolites including enniatins, fusarins, chlamydosporols, JM-47 and others.

MIBiG 4.0: advancing biosynthetic gene cluster curation through global collaboration.

Specialized or secondary metabolites are small molecules of biological origin, often showing potent biological activities with applications in agriculture, engineering and medicine. Usually, the biosynthesis of these natural products is governed by sets of co-regulated and physically clustered genes known as biosynthetic gene clusters (BGCs). To share information about BGCs in a standardized and machine-readable way, the Minimum Information about a Biosynthetic Gene cluster (MIBiG) data standard and repository was initiated in 2015.

Pathogenicity and Metabolomic Characterization of and Challenge in Barley under Controlled Conditions.

Barley is the third most important cereal crop in terms of production in Canada, and Fusarium head blight (FHB) is one of the main fungal diseases of barley. FHB is caused by a species complex of Fusaria, of which Schwabe is the main causal species of FHB epidemics in Canada. Field surveys show that two or more species often co-exist within the same field or grain sample, and is reported as another important species in barley.

Debunking the Myth of T-2/HT-2 Toxin Production.

is commonly detected in field surveys of head blight (FHB) of cereal crops and can produce a range of trichothecene mycotoxins. Although experimentally validated reports of strains producing T-2/HT-2 trichothecenes are rare, is frequently generalized in the literature as a producer of T-2/HT-2 toxins due to a single study from 2004 in which T-2/HT-2 toxins were detected at low levels from six out of forty-nine strains examined. To validate/substantiate the observations reported from the 2004 study, the producing strains were acquired and phylogenetically confirmed to be correctly assigned as ; however, no evidence of T-2/HT-2 toxin production was observed from axenic cultures.

Chromosome-level draft genome sequences of three isolates of the toxigenic fungus showing structural rearrangements.

The whole genomes of three strains were sequenced using Oxford Nanopore Technologies' MinION and assembled into complete, chromosome-level assemblies. The genome consists of eight conserved chromosomes, with evidence of inter-chromosomal structural rearrangements between strains.

CRISPR-Cas9 Gene Editing and Secondary Metabolite Screening Confirm C16 Biosynthetic Gene Cluster Products as Decalin-Containing Diterpenoid Pyrones.

is a causal organism of Fusarium head blight in cereals and maize. Although a few secondary metabolites produced by are considered disease virulence factors, many molecular products of biosynthetic gene clusters expressed by during infection and their associated role in the disease are unknown. In particular, the predicted meroterpenoid products of the biosynthetic gene cluster historically designated as "C16" are likely associated with pathogenicity.

Untargeted metabolomics screening reveals unique secondary metabolite production from section .

section is comprised of many species that infect a broad diversity of important crop plants and cause post-harvest spoilage. section species, such as and , are prolific producers of secondary metabolites that act as virulence factors of disease and are mycotoxins that accumulate in infected tissues-metabolites that can vary in their spectrum of production between individuals from the same fungal species. Untargeted metabolomics profiling of secondary metabolite production using mass spectrometry is an effective means to detect phenotypic anomalies in secondary metabolism within a species.

MIBiG 3.0: a community-driven effort to annotate experimentally validated biosynthetic gene clusters.

With an ever-increasing amount of (meta)genomic data being deposited in sequence databases, (meta)genome mining for natural product biosynthetic pathways occupies a critical role in the discovery of novel pharmaceutical drugs, crop protection agents and biomaterials. The genes that encode these pathways are often organised into biosynthetic gene clusters (BGCs). In 2015, we defined the Minimum Information about a Biosynthetic Gene cluster (MIBiG): a standardised data format that describes the minimally required information to uniquely characterise a BGC.

Untargeted Metabolomic Profiling of Fungal Species Populations.

This chapter describes protocols for the development of consensus chemical phenotypes or "metabolomes" of fungal populations using ultra-high pressure liquid chromatography coupled to high resolution mass spectrometry (UPLC-HRMS). Isolates are cultured using multiple media conditions to elicit the expression of diverse secondary metabolite biosynthetic gene clusters. The mycelium and spent culture media are extracted using organic solvents and profiled by ultra-high pressure chromatography coupled with a high resolution Thermo Orbitrap XL mass spectrometer with the ability to trap and fragment ions to general MS spectra.

Evolution of the Ergot Alkaloid Biosynthetic Gene Cluster Results in Divergent Mycotoxin Profiles in Sclerotia.

Research into ergot alkaloid production in major cereal cash crops is crucial for furthering our understanding of the potential toxicological impacts of upon Canadian agriculture and to ensure consumer safety. An untargeted metabolomics approach profiling extracts of sclerotia from four different grain crops separated the strains into two distinct metabolomic classes based on ergot alkaloid content. Variances in alkaloid profiles were correlated to genetic differences within the gene of the ergot alkaloid biosynthetic gene cluster from previously published genomes and from newly sequenced, long-read genome assemblies of Canadian strains.

A metabolomic study of vegetative incompatibility in Cryphonectria parasitica.

Vegetative incompatibility (VI) is a form of non-self allorecognition in filamentous fungi that restricts conspecific hyphal fusion and the formation of heterokaryons. In the chestnut pathogenic fungus, Cryphonectria parasitica, VI is controlled by six vic loci and has been of particular interest because it impedes the spread of hypoviruses and thus biocontrol strategies. We use nuclear magnetic resonance and high-resolution mass spectrometry to characterize alterations in the metabolome of C.

Apicidin biosynthesis is linked to accessory chromosomes in Fusarium poae isolates.

Background: Fusarium head blight is a disease of global concern that reduces crop yields and renders grains unfit for consumption due to mycotoxin contamination. Fusarium poae is frequently associated with cereal crops showing symptoms of Fusarium head blight. While previous studies have shown F.

Accessory Chromosome-Acquired Secondary Metabolism in Plant Pathogenic Fungi: The Evolution of Biotrophs Into Host-Specific Pathogens.

Accessory chromosomes are strain- or pathotype-specific chromosomes that exist in addition to the core chromosomes of a species and are generally not considered essential to the survival of the organism. Among pathogenic fungal species, accessory chromosomes harbor pathogenicity or virulence factor genes, several of which are known to encode for secondary metabolites that are involved in plant tissue invasion. Accessory chromosomes are of particular interest due to their capacity for horizontal transfer between strains and their dynamic "crosstalk" with core chromosomes.

Transcriptome analysis implicates secondary metabolite production, redox reactions, and programmed cell death during allorecognition in Cryphonectria parasitica.

The underlying molecular mechanisms of programmed cell death associated with fungal allorecognition, a form of innate immunity, remain largely unknown. In this study, transcriptome analysis was used to infer mechanisms activated during barrage formation in vic3-incompatible strains of Cryphonectria parasitica, the chestnut blight fungus. Pronounced differential expression occurred in barraging strains of genes involved in mating pheromone (mf2-1, mf2-2), secondary metabolite production, detoxification (including oxidative stress), apoptosis-related, RNA interference, and HET-domain genes.

The effect of curve running on distal limb kinematics in the Thoroughbred racehorse.

During racing, injury is more likely to occur on a bend than on a straight segment of track. This study aimed to quantify the effects of galloping at training speeds on large radius curves on stride parameters and limb lean angle in order to assess estimated consequences for limb loading. Seven Thoroughbred horses were equipped with a sacrum-mounted inertial measurement unit with an integrated GPS, two hoof-mounted accelerometers and retro-reflective markers on the forelimbs.

Water-jet-assisted liposuction for the treatment of lipedema: Standardized treatment protocol and results of 63 patients.

Background: Lipedema is a condition of painful increase in subcutaneous fat affecting almost exclusively women. Several studies have examined the effectiveness of liposuction in the treatment of lipedema, but none has focused on water-jet-assisted liposuction technique.

Methods: A standardized treatment protocol for liposuction in lipedema, which was established over the course of 15 years, is presented.

Transcriptomic and Exometabolomic Profiling Reveals Antagonistic and Defensive Modes of Action Against .

The mycoparasite ACM941 is under development as a biocontrol organism against , the causative agent of Fusarium head blight in cereals. To identify molecular factors associated with this interaction, the transcriptomic and exometabolomic profiles of and GZ3639 were compared during coculture. Prior to physical contact, the antagonistic activity of correlated with a response heavily dominated by upregulation of polyketide synthase gene clusters, consistent with the detected accumulation of corresponding secondary metabolite products.

Enniatin Production Influences Virulence on Potato Tubers, but not on Durum Wheat or Peas.

is a generalist pathogen responsible for diseases in numerous crop species. The fungus produces a series of mycotoxins including the cyclohexadepsipeptide enniatins. Mycotoxins can be pathogenicity and virulence factors in various plant-pathogen interactions, and enniatins have been shown to influence aggressiveness on potato tubers.

Assessment of clinical and computed tomographic findings for association with the outcome of intraoral cheek tooth extraction in horses and ponies.

Objective: To describe clinical and CT findings for horses and ponies undergoing intraoral cheek tooth extraction and assess potential associations between these features and outcome of the procedure.

Animals: 74 horses and 7 ponies.

Procedures: Medical records were searched to identify horses and ponies that underwent CT and intraoral extraction of ≥ 1 cheek tooth with standing sedation.

The Effect of Training on Stride Duration in a Cohort of Two-Year-Old and Three-Year-Old Thoroughbred Racehorses.

Objective gait monitoring is increasingly accessible to trainers. A more comprehensive understanding of 'normal' gait adaptations is required. Forty two-year-old thoroughbred racehorses were recruited when entering training and followed for 22 months.

Maxillary nerve blocks in horses: an experimental comparison of surface landmark and ultrasound-guided techniques.

Objective: The aim of this preliminary proof-of-concept study was to evaluate and compare the success and complication rate of infiltration of the maxillary nerve of cadaver heads using previously described surface landmarks, standard ultrasound and a novel needle guidance positioning ultrasound system (SonixGPS).

Study Design: Prospective, anatomical, method-comparison study.

Animals: Thirty-eight equine cadaver heads.

Imaging quasiperiodic electronic states in a synthetic Penrose tiling.

Quasicrystals possess long-range order but lack the translational symmetry of crystalline solids. In solid state physics, periodicity is one of the fundamental properties that prescribes the electronic band structure in crystals. In the absence of periodicity and the presence of quasicrystalline order, the ways that electronic states change remain a mystery.

Contribution of industry funded post-marketing studies to drug safety: survey of notifications submitted to regulatory agencies.

Objectives:  To investigate the practice of post-marketing studies in Germany during a three year period and to evaluate whether these trials meet the aims specified in the German Medicinal Products Act.

Design:  Survey of notifications submitted to German regulatory agencies before post-marketing studies were carried out, 2008-10.

Setting:  Notifications obtained through freedom of information requests to the three authorities responsible for registering post-marketing studies in Germany.

IMAGING DIAGNOSIS-IDIOPATHIC HYOID MALFORMATION IN AN 8-YEAR-OLD WELSH SECTION D HORSE.

An 8-year-old gelding was evaluated for intermittent dorsal displacement of the soft palate (DDSP) at exercise. Standing laterolateral radiographs demonstrated an abnormal crossed appearance of the stylohyoid bones. Computed tomography (CT) confirmed abnormal positioning of the left stylohyoid bone and articulations with both the left thyrohyoid and ceratohyoid bones.

Cheek Tooth Extraction Via a Minimally Invasive Transbuccal Approach and Intradental Screw Placement in 54 Equids.

Objective: To describe (1) preoperative findings and surgical technique, (2) intraoperative difficulties, and (3) postoperative complications and long-term outcome of equine cheek tooth extraction using a minimally invasive transbuccal screw extraction (MITSE) technique.

Study Design: Retrospective case series.

Animals: Fifty-four equids; 50 horses, 3 ponies, and 1 mule.