Nutrient stress can have opposite effects on the ability of plants to tolerate foliar herbivory and floral herbivory.

Discovering how organisms respond to the combinations of stressors they face in their environment is an enduring challenge for ecologists. A particular focus has been how natural enemies and abiotic stressors faced by plants may interact in their effect on the ecology and evolution of plant defense strategies. Here, we report on the results of an experiment measuring how reproduction in the clonal herbaceous plant horsenettle (Solanum carolinense) is affected by damage by leaf-feeding and by flower-feeding herbivores-as well as how horsenettle's tolerance of these different types of herbivory may be altered by nutrient stress. Leaf herbivory by lace bugs reduced horsenettle's seed production and root growth, and the relative impacts were greater in fertilized than in nutrient-stressed plants. In contrast, simulated-floral herbivory reduced seed production to a similar degree in fertilized and nutrient-stressed plants. However, compensation for floral herbivory through increased root growth occurred to a much greater extent in the fertilized than in the nutrient-stressed plants. These results can be explained in terms of the limiting resource model of plant tolerance, with leaf damage interpreted as exacerbating carbon limitation in the fertilized plants and floral damage ameliorating carbon limitation in the fertilized plants. These results can be extended to predicting patterns in the field: Although plants in a nutrient-poor environment may have overall low fitness, they are likely to be more tolerant of leaf herbivores-though this benefit may be countered by lower tolerance of any floral herbivores that share the environment.

Plant susceptibility to a shared herbivore is reduced by belowground competition with neighbors.

Spatial variation in plant community composition is an important driver of variation in susceptibility to herbivores. In close proximity, certain neighbors can attract or repel herbivores to a focal plant ("associational effects"). Neighboring plants may also compete for resources, modifying their phenotype in ways that affect susceptibility to herbivores. To test whether and how competition contributes to associational effects, we manipulated the sharing of belowground resources among plant neighbors (spotted Joe Pye weed and common boneset) that serve as alternate hosts for an herbivorous beetle. In the field, the beetle Ophraella notata laid more eggs and inflicted more damage on plants of both species that were released from belowground competition with neighbors. Competition also weakened the effects of neighbor identity during field trials, reducing associational susceptibility. When beetles were forced to choose between the two host species in cage trials, competition again reduced beetle use of Joe Pye weed as a secondary host. To test the role of plant traits related to herbivore defense and nutrition, we quantified leaf protein, specific leaf area, and trichomes, and conducted behavioral assays on leaf disks. Beetles did not distinguish between Joe Pye weed treatments at the leaf disk level, and competition did not impact specific leaf area and protein. Trichome density was higher in both species in the preferred treatment. Overall, our results suggest that belowground interactions between plants may mediate the strength of associational effects, as secondary hosts become more attractive when released from competition with primary host plants.

Myocardial Iba1, MHC class II, and CD3 are diffusely increased in canine myocarditis: A step toward antemortem myocarditis diagnostics.

Canine myocarditis is a rare but serious health concern, potentially causing heart failure and death. Antemortem diagnosis is hampered by the numerous causes, nonspecific course, and dearth of diagnostic criteria. Currently, definitive diagnosis can only be made after death. The current human diagnostic gold standard is endomyocardial biopsy pairing cardiac histopathology with immunohistology to enhance detection of often-multifocal disease. We evaluated immune response markers in the canine heart to establish similar immunohistologic criteria. We hypothesized that myocardial major histocompatibility complex class II (MHCII), cluster of differentiation 3 (CD3), and ionized calcium binding adapter molecule 1 (Iba1), markers increased in human myocarditis, would be increased in canine myocarditis cases. Archived paraffin-embedded myocardial tissue from 22 histopathologically confirmed cases of adult and juvenile myocarditis and 23 controls was analyzed by immunohistochemistry for MHCII, CD3, and Iba1, and the fraction of myocardium with labeling was determined. All 3 markers were significantly increased compared with controls across the entire section: Iba1, 10.1× (P < .0001, Mann-Whitney U test); MHCII, 3.04× (P = .0019); and CD3, 4.4× (P = .0104). To mimic off-target biopsy, samples from 2 mm2 outside of inflammatory foci were analyzed, and these showed significant increases in Iba1 by 3.2× (P = .0036, Mann-Whitney U test) and CD3 by 1.2× (P = .0026). These data show diffusely increased immune response markers with canine myocarditis, with detection potentially independent of tissue sampling. Thus, endomyocardial biopsy and immunohistochemical detection of MHCII, CD3, and Iba1 may permit sensitive antemortem diagnosis of canine myocarditis.

Landscape-dependent effects of varietal mixtures on insect pest control and implications for farmer profits.

Intraspecific plant diversity can significantly impact insect herbivore populations in natural systems. Yet, its role as an insect pest control strategy in agriculture has received less attention, and little is known about which crop traits are important to herbivores in different landscape contexts. Moreover, empirical economic analyses on the cost-effectiveness of varietal mixtures are lacking. We used varietal mixtures of Brassica oleracea crops on working farms to examine how two metrics of intraspecific crop diversity, varietal richness and number of plant colors (color richness), affect crop damage and the incidence and abundance of two insect pest species: Pieris rapae and Phyllotreta spp. We evaluated the context-dependency of varietal mixtures by sampling early- and late-season plantings of B. oleracea crops in farms across a gradient of landscape composition. We developed crop budgets and used a net present value analysis to assess the impact of varietal mixtures on input and labor costs, crop revenues, and profit. We found context-dependent effects of varietal mixtures on both pests. In early-season plantings, color richness did not affect Phyllotreta spp. populations. However, increasing varietal richness reduced Phyllotreta spp. incidence in simple landscapes dominated by cropland, but this trend was reversed in complex landscapes dominated by natural habitats. In late-season plantings, color richness reduced the incidence and abundance of P. rapae larvae, but only in complex landscapes where their populations were highest. Varietal richness had the same effect on P. rapae larvae as color richness. Unexpectedly, we consistently found lower pest pressure and reduced crop damage in simple landscapes. Although varietal mixtures did not affect crop damage, increasing color richness corresponded with increased profits, due to increased revenue and a marginal reduction in labor and input costs. We demonstrate varietal mixtures can significantly impact pest populations, and this effect can be mediated by intraspecific variation in crop color. However, the strength and direction of these effects vary by season, landscape composition, and pest species. The association between varietal color richness and profitability indicates farmers could design mixtures to enhance economic returns. We recommend additional research on the benefits of intraspecific trait variation for farmers.

Genetic impairments in folate enzymes increase dependence on dietary choline for phosphatidylcholine production at the expense of betaine synthesis.

Although single nucleotide polymorphisms (SNPs) in folate-mediated pathways predict susceptibility to choline deficiency during severe choline deprivation, it is unknown if effects persist at recommended intakes. Thus, we used stable isotope liquid chromatography-mass spectrometry (LC-MS) methodology to examine the impact of candidate SNPs on choline metabolism in a long-term, randomized, controlled feeding trial among pregnant, lactating, and nonpregnant (NP) women consuming 480 or 930 mg/d choline (22% as choline-d9, with d9 indicating a deuterated trimethyl amine group) and meeting folate-intake recommendations. Variants impairing folate metabolism, methylenetetrahydrofolate reductase (MTHFR) rs1801133, methionine synthase (MTR) rs1805087 [wild-type (WT)], MTR reductase (MTRR) rs1801394, and methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase-formyltetrahydrofolate synthetase (MTHFD1) rs2236225, influenced choline dynamics, frequently through interactions with reproductive state and choline intake, with fewer genotypic alterations observed among pregnant women. Women with these variants partitioned more dietary choline toward phosphatidylcholine (PC) biosynthesis via the cytidine diphosphate (CDP)-choline pathway at the expense of betaine synthesis even when use of betaine as a methyl donor was increased. Choline intakes of 930 mg/d restored partitioning of dietary choline between betaine and CDP-PC among NP (MTHFR rs1801133 and MTR rs1805087 WT) and lactating (MTHFD1 rs2236225) women with risk genotypes. Overall, our findings indicate that loss-of-function variants in folate-metabolizing enzymes strain cellular PC production, possibly via impaired folate-dependent phosphatidylethanolamine-N-methyltransferase (PEMT)-PC synthesis, and suggest that women with these risk genotypes may benefit from choline intakes exceeding current recommendations.-Ganz, A. B., Shields, K., Fomin, V. G., Lopez, Y. S., Mohan, S., Lovesky, J., Chuang, J. C., Ganti, A., Carrier, B., Yan, J., Taeswuan, S., Cohen, V. V., Swersky, C. C., Stover, J. A., Vitiello, G. A., Malysheva, O. V., Mudrak, E., Caudill, M. A. Genetic impairments in folate enzymes increase dependence on dietary choline for phosphatidylcholine production at the expense of betaine synthesis.

Escherichia coli transfer from simulated wildlife feces to lettuce during foliar irrigation: A field study in the Northeastern United States.

Wildlife intrusion has been associated with pathogen contamination of produce. However, few studies have examined pathogen transfer from wildlife feces to pre-harvest produce. This study was performed to calculate transfer coefficients for Escherichia coli from simulated wildlife feces to field-grown lettuce during irrigation. Rabbit feces inoculated with a 3-strain cocktail of non-pathogenic E. coli were placed in a lettuce field 2.5-72 h before irrigation. Following irrigation, the E. coli concentration on the lettuce was determined. After exclusion of an outlier with high E. coli levels (Most Probable Number = 5.94*108), the average percent of E. coli in the feces that transferred to intact lettuce heads was 0.0267% (Standard Error [SE] = 0.0172). Log-linear regression showed that significantly more E. coli transferred to outer leaves compared to inner leaves (Effect = 1.3; 95% Confidence Interval = 0.4, 2.1). Additionally, the percent of E. coli that transferred from the feces to the lettuce decreased significantly with time after fecal placement, and as the distance between the lettuce and the feces, and the lettuce and the sprinklers increased. These findings provide key data that may be used in future quantitative risk assessments to identify potential intervention strategies for reducing food safety risks associated with fresh produce.

Assessing the impact of fire on the spatial distribution of Larrea tridentata in the Sonoran Desert, USA.

In southwestern American deserts, fire has been historically uncommon because of insufficient continuity of fuel for spreading. However, deserts have been invaded by exotic species that now connect the empty space between shrubs to carry fire. We hypothesized that fire would change the spatial distribution of surviving Larrea tridentata shrubs. We established two study plots, one each in a burned and unburned area, and recorded location and living status of all shrubs. We performed univariate and bivariate point pattern analyses to characterize the impact of fire on the overall distribution of shrubs. Additionally, we used a simple wildfire model to determine how close we could come to reconstructing the observed spatial pattern of living and dead shrubs. We found a hyper-dispersed pattern of shrubs at finer scales and a random pattern at broader scales for both the unburned plot and for the living and dead shrubs combined in the burned plot, the latter providing an approximation of the pre-burn distribution of shrubs. After fire, living shrubs showed a clustered pattern at scales >2.5 m, whereas dead shrubs were randomly distributed, indicating that fire caused a change in the spatial pattern of the surviving shrubs. The fire model was able to partially reconstruct the spatial pattern of Larrea, but created a more clustered distribution for both living and dead shrubs. Our study reinforces the key role of fire in altering landscapes that had not been habituated to fire, and suggests the existence of potential cascading effects across the entire plant community.

In-gallery social behaviors of the ambrosia beetle, Xylosandrus germanus (Coleoptera: Curculionidae).

The east Asian ambrosia beetle Xylosandrus germanus (Blanford) was first detected in the United States in 1932. It now occurs across much of eastern North America and parts of the Pacific Northwest. It attacks a broad range of stressed, woody hosts including ornamental and orchard species. The foundress tunnels into the sapwood of hosts where it cultures a symbiotic fungus as food for its offspring. A few other ambrosia beetles have been shown to possess a facultatively eusocial structure among gallery members, but this has not been described for Xylosandrus spp. Using a novel artificial diet arena, we quantified the behaviors of X. germanus larvae and adults (foundress and mature offspring) over 10 wk inside their galleries. Foundresses were responsible for constructing the gallery. They also initially tended the fungal garden and brood but eventually spent most of their time blocking the gallery entrance. Larvae were mainly observed to feed, crawl, or be inactive within the gallery, regardless of the absence or presence of adult siblings. Adult female offspring were primarily inactive, likely due to dormancy. Adult male offspring actively crawled and attempted to mate with their sisters before eventually dispersing out of the gallery. Cooperative hygienic behaviors (removal of frass, cannibalism of dead nest mates, grooming siblings) were observed but a division of labor among offspring was not clear. Rather, foundress behaviors were mostly distinct from offspring behaviors, particularly as the gallery aged. Because no overlap in generations occurred, X. germanus displays a quasisocial structure.

Complex Interactions Between Weather, and Microbial and Physicochemical Water Quality Impact the Likelihood of Detecting Foodborne Pathogens in Agricultural Water.

Agricultural water is an important source of foodborne pathogens on produce farms. Managing water-associated risks does not lend itself to one-size-fits-all approaches due to the heterogeneous nature of freshwater environments. To improve our ability to develop location-specific risk management practices, a study was conducted in two produce-growing regions to (i) characterize the relationship between Escherichia coli levels and pathogen presence in agricultural water, and (ii) identify environmental factors associated with pathogen detection. Three AZ and six NY waterways were sampled longitudinally using 10-L grab samples (GS) and 24-h Moore swabs (MS). Regression showed that the likelihood of Salmonella detection (Odds Ratio [OR] = 2.18), and eaeA-stx codetection (OR = 6.49) was significantly greater for MS compared to GS, while the likelihood of detecting L. monocytogenes was not. Regression also showed that eaeA-stx codetection in AZ (OR = 50.2) and NY (OR = 18.4), and Salmonella detection in AZ (OR = 4.4) were significantly associated with E. coli levels, while Salmonella detection in NY was not. Random forest analysis indicated that interactions between environmental factors (e.g., rainfall, temperature, turbidity) (i) were associated with likelihood of pathogen detection and (ii) mediated the relationship between E. coli levels and likelihood of pathogen detection. Our findings suggest that (i) environmental heterogeneity, including interactions between factors, affects microbial water quality, and (ii) E. coli levels alone may not be a suitable indicator of food safety risks. Instead, targeted methods that utilize environmental and microbial data (e.g., models that use turbidity and E. coli levels to predict when there is a high or low risk of surface water being contaminated by pathogens) are needed to assess and mitigate the food safety risks associated with preharvest water use. By identifying environmental factors associated with an increased likelihood of detecting pathogens in agricultural water, this study provides information that (i) can be used to assess when pathogen contamination of agricultural water is likely to occur, and (ii) facilitate development of targeted interventions for individual water sources, providing an alternative to existing one-size-fits-all approaches.

Youth Willingness to Purchase Whole Grain Snack Packs from New York City Corner Stores Participating in a Healthy Retail Program.

Corners stores in low-income communities are a promising setting to intervene in youth whole grain intake. One strategy that may encourage whole grain intake is if corner stores were to pair and sell whole grain snacks in combination with either a liked fruit or vegetable and an optional condiment (i.e., a whole grain snack pack). This study examined youth in terms of their (1) liking of fruits, vegetables, and whole grain snacks; (2) perceptions about which fruits and vegetables pair best with whole grain snacks; and (3) willingness to pay for a whole grain snack pack. One-time intercept surveys were conducted with 10-18-year-olds (n = 402) who visited a New York City (NYC) corner store (n = 34) participating in the City Harvest Healthy Retail Program. On average, youth were willing to spend $2.38 (SD $4.32) on a whole grain snack pack. Higher overall liking scores for vegetables and whole grain snacks were associated with willingness to spend 24.4% (95% confidence interval (CI): 11.5-38.7%) and 21.6% (95%CI: 5.2-40.6%) more on whole grain snack packs, respectively. In conclusion, youth are receptive to purchasing whole grain snack packs from NYC corner stores participating in a healthy retail program.