Differential Stem Proteomics and Metabolomics Profiles for Four Wheat Cultivars in Response to the Insect Pest Wheat Stem Sawfly.

Common wheat (Triticum aestivum L.) is a global staple crop, and insect pests can impact grain yield. The wheat stem sawfly (Cephus cinctus, WSS) is a major wheat pest, and while partial resistance has been deployed by breeding for a solid-stem trait, this trait is affected by environment. Here, a proteomics and metabolomics study was performed on four wheat cultivars to characterize a molecular response to WSS infestation. The cultivars Hatcher (hollow-stem partially tolerant), Conan (semisolid-stem-resistant), and Denali and Reeder (hollow-stem-susceptible) were infested with WSS, and changes in stem proteins and metabolites were characterized using liquid chromatography-mass spectrometry. The proteome was characterized as 1830 proteins that included five major biological processes, including metabolic processes and response to stimuli, and the metabolome (1823 metabolites) spanned eight chemical superclasses, including alkaloids, benzenoids, and lipids. All four varieties had a molecular response to WSS following infestation. Hatcher had the most distinct changes, whereby 62 proteins and 29 metabolites varied in metabolic pathways involving enzymatic detoxification, proteinase inhibition, and antiherbivory compound production via benzoxazinoids, neolignans, and phenolics. Taken together, these data demonstrate variation in the wheat stem molecular response to WSS infestation and support breeding for molecular resistance in hollow-stem cultivars.

GC-MS Metabolomics to Evaluate the Composition of Plant Cuticular Waxes for Four Triticum aestivum Cultivars.

Wheat (Triticum aestivum L.) is an important food crop, and biotic and abiotic stresses significantly impact grain yield. Wheat leaf and stem surface waxes are associated with traits of biological importance, including stress resistance. Past studies have characterized the composition of wheat cuticular waxes, however protocols can be relatively low-throughput and narrow in the range of metabolites detected. Here, gas chromatography-mass spectrometry (GC-MS) metabolomics methods were utilized to provide a comprehensive characterization of the chemical composition of cuticular waxes in wheat leaves and stems. Further, waxes from four wheat cultivars were assayed to evaluate the potential for GC-MS metabolomics to describe wax composition attributed to differences in wheat genotype. A total of 263 putative compounds were detected and included 58 wax compounds that can be classified (e.g., alkanes and fatty acids). Many of the detected wax metabolites have known associations to important biological functions. Principal component analysis and ANOVA were used to evaluate metabolite distribution, which was attributed to both tissue type (leaf, stem) and cultivar differences. Leaves contained more primary alcohols than stems such as 6-methylheptacosan-1-ol and octacosan-1-ol. The metabolite data were validated using scanning electron microscopy of epicuticular wax crystals which detected wax tubules and platelets. Conan was the only cultivar to display alcohol-associated platelet-shaped crystals on its abaxial leaf surface. Taken together, application of GC-MS metabolomics enabled the characterization of cuticular wax content in wheat tissues and provided relative quantitative comparisons among sample types, thus contributing to the understanding of wax composition associated with important phenotypic traits in a major crop.

On-farm harvest timing effects on alfalfa weevil across the Intermountain West region of the United States.

Alfalfa (Medicago sativa L.) is an economically important commodity in the Intermountain Western United States. A major concern for alfalfa producers in this region is the alfalfa weevil (Hypera postica Gyllenhal). Insecticide resistance development coupled with regulatory changes in pesticide use has resulted in renewed interest by producers in non-chemical control methods such as cultural control. One such cultural control method is early harvest, which consists of producers timing their harvests early in the season to decrease alfalfa weevil damage. This method is thought to be effective by exposing weevil larvae to adverse conditions before significant damage occurs. Still, early harvest can be difficult to employ because recommendations are often vague. To better understand how early harvest impacts both alfalfa weevils and their natural enemies and how producers are using this method across the Intermountain Western United States, we conducted a study in alfalfa production fields in Colorado, Montana, and Wyoming over three growing seasons. We determined that the timing of the initial alfalfa harvest spanned more than 1 month across fields, and alfalfa plant stage at harvest ranged from late vegetative to early bloom. Harvest was more impactful on reducing alfalfa weevil densities the earlier it was implemented. Removing windrows in a timely manner is likely useful to further decrease alfalfa weevil densities. Harvest timing was not associated with parasitism rates of alfalfa weevil, but higher parasitism rates were associated with lower post-harvest alfalfa weevil densities. This work has increased our understanding of early harvest in an on-farm setting and to improve recommendations for producers across the Intermountain Western United States.

Evaluation of outcome associated with feline trauma: A Veterinary Committee on Trauma registry study.

OBJECTIVE: To evaluate outcome (survival to discharge) among trauma types (blunt, penetrating, both) in cats. Secondary objectives were to evaluate for associations between trauma type, injury severity, and the diagnostics and interventions selected by primary clinicians. DESIGN: Retrospective evaluation of veterinary trauma registry data. SETTING: Veterinary Committee on Trauma (VetCOT) veterinary trauma centers (VTCs). ANIMALS: A total of 3895 feline trauma patients entered in the VetCOT trauma registry from April 1, 2017 to December 31, 2019. INTERVENTIONS: Data collected included patient demographics, trauma type, Abdominal Fluid Score (AFS), Animal Trauma Triage (ATT) score, surgical intervention, glide sign on Thoracic Focused Assessment with Sonography for Trauma, Triage, and Tracking (TFAST), pleural effusion on TFAST, modified Glasgow Coma Scale (mGCS), and outcome (survival to discharge). MEASUREMENTS AND MAIN RESULTS: Data from 3895 cats were collected over a 30-month period. Incidence of trauma types was as follows: blunt, 58% (95% confidence interval [CI]: 56%-59%); penetrating, 35% (95% CI: 34%-37%); and combination, 7.4% (95% CI: 6.7%-8.3%). Differences in survival incidence among the trauma types were identified: blunt, 80% (95% CI: 78%-81%); penetrating, 90% (95% CI: 89%-92%); and combined, 68% (95% CI: 63%-74%) (P < 0.01). Cats in the penetrating trauma group had the lowest proportion of severe injuries (6%) and highest proportion of mGCS of 18 (89%); cats with combined trauma had the highest proportion of severe injuries (26%) and lowest proportion of mGCS of 18 (63%). Point-of-care ultrasound and surgery were not performed in the majority of cases. When surgery was performed, the majority of blunt cases' procedures occurred in the operating room (79%), and the majority of penetrating cases' procedures were performed in the emergency room (81%). CONCLUSIONS: Cats suffering from penetrating trauma had the best outcome (survival), lower ATT scores, and higher mGCS overall. Cats that sustained a component of blunt trauma had a lower survival rate, higher ATT scores, and the highest proportion of mGCS <18.

Comparison of clinical outcomes in cases of blunt, penetrating, and combination trauma in dogs: A VetCOT registry study.

OBJECTIVE: To evaluate outcome (survival to discharge) among trauma types (blunt, penetrating, both) in dogs. The secondary objective was to evaluate if other trauma registry parameters differ between trauma types and influence survival. DESIGN: Retrospective evaluation of veterinary trauma registry data. SETTING: Veterinary Committee on Trauma (VetCOT) identified Veterinary Trauma Centers (VTCs). ANIMALS: A total of 20,289 canine trauma patients with data entered in the VetCOT trauma registry from April 1, 2017 to December 31, 2019 INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Data were obtained from the VetCOT database and included patient demographics, trauma type (blunt, penetrating, both), Animal Trauma Triage (ATT) score, modified Glasgow Coma Scale (mGCS), abdominal fluid score (AFS), loss of glide sign on thoracic focused assessment with sonography for trauma (TFAST), pleural effusion on TFAST, surgical procedure performed and in what location (emergency room vs operating room), and outcome. Data from 20,289 dogs were collected over a 30-month period. The most common type of trauma was penetrating (10,816, 53.3%), followed by blunt (8360, 41.2%) and then combined blunt and penetrating trauma (1113, 5.5%). Dogs suffering only penetrating trauma had a 96.5% survival rate, blunt trauma had an 89.5% survival rate, and combined trauma had an 86.3% survival rate. Dogs suffering from both types of trauma had higher ATT scores, lower mGCS scores, and were more likely to be admitted to the ICU. Trauma type, mGCS, and ATT score were found to be associated with survival. CONCLUSIONS: The present study highlights that dogs suffering from a combination of blunt and penetrating trauma are more likely to suffer moderate to severe injuries, have lower survival rates, and are more likely to be admitted to the ICU compared to dogs suffering from only blunt or penetrating trauma. Trauma type, mGCS, and ATT scores were found to be associated with survival in all groups.

Survey of Wheat Stem Sawfly (Hymenoptera: Cephidae) Infesting Wheat in Eastern Colorado.

From 2012 through 2020, a survey of wheat stem sawfly, Cephus cinctus Norton, was conducted in wheat (Triticum aestivum L.) fields in Eastern Colorado. In 2013, results showed sawfly infestations concentrated in the northern part of the state with only a few highly infested sites, with 38 of the 94 sampled sites having any infestation (five of which had >50% infestation levels). By 2020 sawfly had been found in all eastern counties sampled, and 72 of the 106 sites sampled were found to contain sawfly (11 of which had >50% infestation levels). The spread of this pest across the Colorado wheat-growing region will have lasting economic effects. The information gathered from this and future surveys will inform wheat variety development and aid in management decisions made by growers across the state.

Effects of Landscape Composition on Wheat Stem Sawfly (Hymenoptera: Cephidae) and Its Associated Braconid Parasitoids.

Several agroecological and integrated pest management strategies focus on landscape management to increase complexity and foster biodiversity. However, landscape complexity does not always enhance biological control and in some cases may lead to increased pest populations. We examined the prevalence of two Bracon parasitoids, Bracon cephi Gahan and Bracon lissogaster Muesebeck (Hymenoptera: Braconidae), and their host the wheat stem sawfly Cephus cinctus Norton, a major pest of wheat. We assessed the degree of noncrop and crop host plant use and responses to landscape composition. We found no instances of parasitism by either Bracon species in our three-year, statewide winter wheat survey but found small populations of Bracon in noncrop landscapes throughout eastern and western Colorado. We used model selection to examine how local (500 m scale) and landscape (5 km scale) cover of suitable noncrop and crop habitats potentially affects abundances of Bracon and wheat stem sawfly. Our best fit model for wheat stem sawfly suggests that a decrease in noncrop cover at the landscape scale leads to an increase in wheat stem sawfly infestation. Our best fit model for Bracon parasitism suggests an increase in wheat cover at the local level results in the greatest increase in the odds of parasitism by either species of Bracon. Herbaceous cover at local and landscape scales were also significant predictors of Bracon parasitism. The results of this study suggest that pest and natural enemies respond differently to landscape composition and these responses should be evaluated before management decisions are made.

Temperature Limits for the Brown Wheat Mite, in Colorado.

Brown wheat mites, Petrobia latens (Müller 1776, Acari: Tetranychidae), are sporadic yet economically damaging pests of winter cereals. In Colorado, their life history is closely tied to the development of winter wheat, where they are present in the field from crop planting in late September through harvest in early June. In order to withstand winter months, these mites are able to survive cold temperatures. However, the mechanisms of cold hardening and their temperature limits are unknown. This research documents the seasonal supercooling points of the brown wheat mite. Their seasonal average supercooling point stayed consistent throughout the year, never varying more than a degree from the overall average supercooling point of -17°C. The greatest variation in supercooling point was seen in the spring, during which supercooling point temperatures ranged from -9.2 to -25.5°C. We also documented the upper and lower lethal temperatures for the brown wheat mite. When comparing small nymphs to large nymph and adult stages, small nymphs were slightly more cold tolerant (lethal temperature estimates required to kill 99% of the population [LT99] were -30.8 and -30.6°C, respectively), but less heat tolerant (LT99 was 50 and 56°C, respectively).

Host Plants of the Wheat Stem Sawfly (Hymenoptera: Cephidae).

Wheat stem sawfly (Cephus cinctus Norton) is a pest of economic importance across much of the wheat (Triticum aestivum L.)-growing areas of the western Great Plains of North America as well as an ecologically important insect owing to its wide range of grass hosts. Little research has been published regarding the noncultivated native and invasive grasses attacked by this insect. Knowledge of the complete host range of C. cinctus can inform future research about potential new sources of genetic resistance, improve understanding of the biology and spread of natural enemies, and better define this insect's role in grassland and agricultural systems. The aim of this review is to compile a checklist of reported host plants of C. cinctus and present data from an extensive survey of noncultivated hosts used by C. cinctus.

Phylogeography of the Wheat Stem Sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae): Implications for Pest Management.

The wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), is a key pest of wheat in the northern Great Plains of North America, and damage resulting from this species has recently expanded southward. Current pest management practices are inadequate and uncertainty regarding geographic origin, as well as limited data on population structure and dynamics across North America impede progress towards more informed management. We examined the genetic divergence between samples collected in North America and northeastern Asia, the assumed native range of C. cinctus using two mitochondrial regions (COI and 16S). Subsequently, we characterized the structure of genetic diversity in the main wheat producing areas in North America using a combination of mtDNA marker and microsatellites in samples collected both in wheat fields and in grasses in wildlands. The strong genetic divergence observed between North American samples and Asian congeners, in particular the synonimized C. hyalinatus, did not support the hypothesis of a recent American colonization by C. cinctus. Furthermore, the relatively high genetic diversity both with mtDNA and microsatellite markers offered additional evidence in favor of the native American origin of this pest. The genetic diversity of North American populations is structured into three genetic clusters and these are highly correlated with geography. Regarding the recent southern outbreaks in North America, the results tend to exclude the hypothesis of recent movement of damaging wheat stem sawfly populations from the northern area. The shift in host plant use by local populations appears to be the most likely scenario. Finally, the significance of these findings is discussed in the context of pest management.