Horticulture

The magnitude of the potential acidity of the soil depends on the type and amount of clay and organic matter. Although there is some inverse parallelism between acidity saturation and base saturation, these terms are not complementary because they have different denominators . Contrary to Al toxicity and acidity saturation, there is no direct association between base saturation and crop yields. Instead, a minimum base saturation threshold is defined such that, above it, no soil acidity problems are detected . Therefore, recommended target base saturation levels must be defined locally for each crop type . Most lime requirement methods used in temperate regions target soil pH , estimating with locally-calibrated models the lime rate required to raise the soil pH to a specific level . In acid tropical soils, maximum yields can be obtained with a pH as low as 5, depending on other soil chemical properties , and raising the pH to higher values can result in a loss of soil structure and other problems . Therefore, a target pH level is seldomly used, and when used, it must be defined locally . There is a negative exponential relationship between exchangeable acidity and soil pH . Very high exchangeable acidity values are only found in soils with a low pH,garden grow bags but not all soils with a low pH have high exchangeable acidity.

Exchangeable acidity approaches 0 at a pH above 5.5, and there is virtually no exchangeable acidity at a pH above 6 . Therefore, a target pH between 5.5 and 6 should be high enough to address Al toxicity problems.Acid tropical soils have very low plant-available phosphorus because of the high P fixation capacity of Fe and Al oxides often present in weathered tropical soils. Liming has the associated benefit of increasing P availability, which might result in significant yield responses, particularly when P fertilization is low . However, liming can only provide short-term relief to P deficiencies in soils with low P reserves . Therefore, phosphorus availability is not considered a direct target of liming, and lime requirement models do not consider it. However, the increase in P availability can be an important reason for observing a yield response to lime.The lime recommendation models were evaluated using data from four soil incubation studies that measured the effect of liming on exchangeable acidity and ECEC or acidity saturation . Studies only measuring the effect of liming on pH were not considered. Soil incubation studies are experiments in which soil samples are mixed with different lime treatments and incubated under controlled conditions for about a month to ensure that all lime reacts with the soil. The liming effect is assessed by measuring chemical soil properties before and after each treatment. The data from Kamprath , Cochrane et al. , and Ananthacumaraswamy and Baker were readily available, but Teixeira et al. soil data were not. The Teixeira et al. data was reconstructed in two steps: the initial soil properties were back solved from lime requirement formulas and lime rates, and the final soil properties were estimated using the regression formulas provided in the supplementary information .

Table 1 describes the main features of these four data sets. We have shown important differences in lime rates and prediction accuracy depending on the target soil property and model . When the target is to ameliorate the Al toxicity of the soil by neutralizing its acidity saturation to a certain level, both Kamprath and Cochrane et al. models provided reasonable accuracy . Nevertheless, the new model formulated in Eq. 12 offers improved accuracy and the advantage of being sustained by a formal mathematical derivation that can be expanded . Similarly, the base saturation model also has great prediction accuracy, particularly for target base saturation levels of around 50% . In contrast, no model based on a target pH can deliver accurate results without additional soil tests, and they need to be developed locally . The model presented here is the only model based on a target acidity saturation with greater accuracy than the original Cochrane et al. model . The authors of the ACID4, NuMaSS, and MG5 models claimed that they modified the Cochrane et al. model to improve the accuracy for their target region. However, there are no available data supporting those statements. The decreased accuracy that we found may partly be because we did not have access to these data. However, we believe that these more complex models suffer from overfitting to the datasets used to build them. In other words, they may perform better in particular regions, but this has come at the expense of general validity. Conversely, the new model is more robust than previous models because it is based on mathematical foundations and strong empirical relations. These relations are consistent through a wide range of soils from different regions.

However, we observed a small incubation study effect in the relations shown in Figure 3, which might be a consequence of the soil region or, more likely, because of the incubation study per se . Experimental results have an error component, including systematic errors that are consistent within one experiment but differ between experiments, introducing statistical bias. This bias can be reduced with standardized procedures. However, lime incubation studies are not fully standardized, and they differ in the incubation time and temperature, liming materials, and water additions, among other variables. For instance, we excluded data from an incubation study that used tap water rather than distilled water to keep the soil samples moist during the incubation because control treatments had significantly more exchangeable Ca2+ and less exchangeable acidity than the initial conditions. A more thorough standardization of experimental procedures for measuring liming effects would help the development of general models for lime requirement estimation. A novel feature of the model formulated in Eq. 12 is that the lime factor is a continuous function of TAS. The Cochrane et al. model modifies the lf depending on TAS and the initial acidity saturation, using a discontinuous rule with two fixed lf . However, the proposed rule does not always improve accuracy, not even in their data, as shown by the points with incorrect lf . In the MG5 and NuMaSS methods, the lf depends on clay content or activity. Our review does not show evidence for a need to adjust the lf as a function of clay, despite the wide range of clay content and soils included in the four soil incubation studies used here. Adjusting the lf and lime rates by clay content might be a work-around to account for differences in soil bulk density when the method returns lime rates in tons per hectare without directly including the soil bulk density in the formulas. Nevertheless, clay type and content could be considered in future corrections of the TAS method, particularly if there are high deviations in the association between lime rate and Δexch. acid and Δexch. bases. It seems counter intuitive that, while both the acidity saturation and base saturation models are highly accurate on their target,tomato grow bags the lime requirement they predict can be so sharply different . These differences highlight the importance of identifying the soil chemical property most associated with the crop yield response to liming. It might be that reaching a given level for some property, such as a base saturation of 70% or a pH of 6, guarantees that all soil acidity problems are solved without leading to over liming problems. However, this approach can also result in lime requirement estimates that are much too high , which might be particularly problematic when lime is expensive, and its manipulation cumbersome. The alternative is to target the most limiting factor for crop yield, which is frequently Al toxicity in acid tropical soils . However, this approach can under predict lime requirements when Al toxicity is the only target but not the acidity problem most limiting crop yields.

A comprehensive approach would predict the lime rate needed to tackle every potential acidity problem while considering other management alternatives. However, crop responses to other acidity problems, such as calcium and magnesium deficiencies, are unclear, and their liming requirements have not been defined. Thus, more research on crop responses to lime in soils with these specific acidity problems is needed to develop a lime requirement method that tackles them all. Lime requirement models can be useful for strategic research on potential lime use in tropical regions where liming is still a rare practice and the experimental evidence is scarce . These models estimate the lime rate needed to reach a target soil condition based on readily available standard soil data . Such information could be used together with the crop response to that soil condition to estimate the effect of liming on crop yield. For instance, there is ample evidence of the association between acidity saturation and crop yields . Therefore, the expected yield response to lime can be predicted by estimating what fraction of the maximum yield is observed at the current acidity saturation level while assuming that the final yield after liming is the inverse of that fraction. If data on lime and grain prices is available, such functions can be used to get a first approximation of the potential profitability of liming. Such analysis can help identify regions where liming investments might be more successful, pinpointing national governments and private sector efforts. However, this does not mean that the readily available soil data used by the models reviewed here has sufficient quality for farm-level recommendations . Therefore, farm-level lime requirements would be more accurate when based on soil properties measurements or additional local soil-quality indicators, such as soil color, soil texture, or presence of specific plant species . The soil properties used by the lime requirement models reviewed here are wet-lab measurements, which are costly and may be inaccessible for farmers in the tropics. Therefore, farmers in the tropics could benefit from cost effective, quick tests for lime requirement prediction, but these need to be developed locally.Worldwide, biodiversity is declining at unprecedented rates, threatening species persistence as well as the benefits humans gain from ecosystems. These benefits, known as ecosystem services, have become an increasingly important argument for biodiversity conservation. The economic and other benefits from ecosystems can motivate conservation action, and are more and more being used in payment for ecosystem service schemes. Once an economic value of the service has been determined, it can be captured in commercial markets or quantified in terms comparable with economic services and manufactured capital. These economic values can then potentially be used to support biodiversity conservation within policies. The use of ecosystem services arguments for justifying biodiversity conservation is, however, not without risk or controversy. Many experimental studies show that biodiversity increases the magnitude and/or stability of ecosystem functioning , and that most species contribute to ecosystem functioning in some way. However, such studies do not consider the costs of maintaining or promoting biodiversity, even though costs are generally a limiting factor for implementing real-world conservation policies. When the economic pay-off from ecosystem services is the main factor motivating conservation, the cost-effective action is to conserve the subset of species that provide the greatest return at relatively short timescales. Because real-world communities are almost invariably dominated by a small number of species that often respond readily to conservation management, we hypothesize that in real-world landscapes the majority of the services is provided by a relatively small number of species; that these species are generally common, and that threatened species rarely contribute to present ecosystem service delivery; and that the most important ecosystem-service-providing species can be easily enhanced by simple management actions that are insufficient to support threatened species. Support for these hypotheses would suggest that delivery of ecosystem services is insufficient as a general argument for biodiversity conservation. Here we test these hypotheses using data from 90 studies and 1,394 crop fields on crop-visiting bee communities from five continents. Pollination is an important ecosystem service. The economic contribution of pollinators to crop production is significant, and bees are generally considered the most important pollinators of crops. We find that wild bee communities contribute on average over $3,000 ha 1 to the production of insect-pollinated crops. However, a limited subset of all known bee species provides the majority of pollination services because, across different crops, years and large bio geographical regions, crop-visiting bee communities are dominated by a small number of common species and rarely contain regionally threatened species.

Besides reactive oxygen species , ATs also inhibit the production of reactive nitrogen species , particularly nitric oxide , as well as their associated oxidative processes. Furthermore, release of pro-inflammatory mediators and adhesion molecules is suppressed by ATs via targeting of the respective signaling pathways, e.g., the arachidonic acid and the tumor necrosis factor -α, nuclear factor -κB pathways. Aside from cardioprotection, the anti-inflammatory property of ATs also contributes to their anti-obesity effect and action in adipose tissue. Studies using AT-rich fruit extracts showed that ATs prevented the upregulation of inflammatory response in adipose tissue, when triggered with consumption of a high-fat diet, by regulating the NF-κB stress signaling pathway. ATs also reduce the level of pro-inflammatory cytokines and modulate the expression of adipocytokines. Although studies using cell lines and animal models support the anti-obesity effect of ATs, a causal relationship between AT consumption and reduction of body mass index has yet to be conclusively established by clinical studies. This could be due to one or more parameters that are associated with different clinical studies,plastic plant pot such as the amount and types of ATs ingested, the food matrix of ATs, and the inherent difference of the study subjects.

In addition to their antioxidant and anti-inflammatory activities, ATs act in cancer chemoprevention by inducing terminal differentiation of tumor cells and thus impeding tumorigenesis. Prevention of malignant cell transformation and inhibition of cancer cell proliferation by ATs have also been reported. ATs can further interfere with cancer development by activating caspases and inducing apoptosis of cancer cells. However, the cancer chemotherapeutic effect of ATs still requires strong supporting evidence from clinical studies. In addition, the bio-conversion of ATs by human intestinal micro-biome should be rigorously investigated. Similar to other AT-rich fruits, the pomegranate also has a cardioprotective effect by targeting two major causal factors of atherosclerotic lesion and cardiovascular disease, namely the accumulation of cholesterol and oxidized lipids, and arterial macrophage foam cell formatio. Pomegranate juice and fruit extracts are also associated with antidiabetic activities, largely due to the antioxidant properties of HTs and ATs in reducing oxidative stress and lipid peroxidation. Supplements of pomegranate juice or fruit extracts at 1 g/kg/day for five weeks, effectively enhanced endothelial NO synthase expression, as well as plasma nitrate and nitrite levels in plasma in obese Zucker rats fed with an atherogenic diet. In addition, supplements of juice at 100 mg/kg/day or 300 mg/kg/day for four weeks prevented the development of high blood pressure in diabetic rats. Consumption of pomegranate juice rich in ATs and HTs also lowered systolic and diastolic blood pressure in hypertensive patients.

The bioactivities of pomegranate ATs have mostly been studied in the context of pomegranate juice and fruit extracts. However, these AT-rich sources are abundant in vitamin C, carotenoids, and HTs, which can also contribute to the ascribed bioactivities. To this end, “white” pomegranate cultivars are available, which lack the accumulation of ATs in leaves, fruits and flowers. Comparative analysis of the white and AT-rich pomegranate cultivars will be informative as to the specific role of ATs in the health-promoting activities of pomegranate juice and fruit extracts. ETs are abundant in pomegranate fruit peel, as well as juice and extracts that are produced commercially from the whole fruit. Besides pomegranate, ETs are also present in a wide range of medicinal and food plants, such as many berries, nuts, and herbs. As such, the knowledge of bio-transformation and bio-conversion of pomegranate ETs by human and microbial enzymes will inform nutritional and pharmacological studies on ETs that are isolated from other plants. EA, a hydrolysis product of ETs , can be detected in the blood circulation of healthy volunteers within half an hour of ingesting pomegranate juice or extracts. The human gut micro-biota can further convert EA into urolithins prior to their absorption by the intestinal cells. It was shown that urolithin concentrations reached up to 18.6 µM in the plasma of healthy volunteers after consuming pomegranate juice for five consecutive days. Aromatase that converts androgens to estrogens has been considered to be a therapeutic target for treating the hormone-sensitive type of breast cancer. Although several urolithins exhibited anti-aromatase activities in a placental microsome-based enzyme assay, only UB competitively inhibited and most effectively suppressed the aromatase activity in an aromatase-over expressing breast cancer cell line.

Consistent with “in cell” aromatase-inhibitory activity, UB also significantly arrested testosterone-induced proliferation of MCF-7aro cells, suggesting an underlying mechanism of aromatase inhibition. Interestingly, estrogen-induced proliferation of MCF-7aro cells was also inhibited by urolithins. However, inhibition was was likely effected through mechanisms independent of the aromatase activity. Besides breast cancer, urolithins can also reduce the risks of another sex steroid hormone-related cancer, endometrial cancer. UA arrested cell cycle at the G2/M phase and regulated the expression of cell cycle-related proteins at this phase. On the other hand, UA was shown to act as an estrogen agonist and modulate the estrogen-receptor α -dependent gene expression in the ER-positive endometrial cancer cells. It remains to be determined whether and how estrogen signaling could be involved in the suppression of endometrial cancer cell growth by urolithins. The canonical Wnt/β-catenin signaling pathway has been shown to activate T-cell factor transcription and function in colon carcinogenesis. To determine the potential involvement of urolithins in colon carcinogenesis through Wnt signaling, phenolic extracts of several fruits rich in ETs, including pomegranate, strawberry , and Jamun berry , as well as pure EA and UA compounds, were tested in the human embryonic kidney 293T cells that express a reconstructed canonical Wnt signaling pathway. Although the fruit extracts and chemicals all inhibited Wnt signaling, only UA exhibited an IC50 value that was physiologically relevant in the lumen of colons when taking enterohepatic circulation of urolithins into consideration. During the initiation stage of the HT-29 colon cancer cells, urolithins A–D inhibited TCDD-induced, CYP1-mediated EROD activities. At the cancer progression stage, UA and UB impaired the proliferation of HT-29 cells and led to cell cycle arrest at the G2/M phase, as well as activation of CDKN1A expression. In addition, treatment with urolithins resulted in activation of caspases 3, 8, and 9, suggesting that urolithins induced both the extrinsic and intrinsic apoptotic pathways in HT-29 cells. UA, UB, and mUA inhibited the proliferation of the T24 human bladder cancer cells in vitro, with IC50 values of 43.9, 35.2, and 46.3 µM, respectively, comparable to the UA inhibition of Wnt signaling at an IC50 of 39 µM. The transcript and protein levels of Phospho-p38 mitogen-activated protein kinase were increased by the urolithin treatment, while those of MAP kinase kinase kinase1 and Phospho-c-Jun were decreased in the T24 cells. Furthermore, these urolithins reduced the level of H2O2-induced oxidative stress and induced apoptosis through activation of caspase 3 and PPAR-γ protein expression. Inflammatory responses involving the activation of neutrophils and monocytes play a central role in the development of cardiovascular diseases. Intriguingly,nursery pots the number of free hydroxyl functional groups on urolithins appeared to impact how they modulate the inflammatory functions of neutrophils.

Among the urolithins tested , UA exhibited the most potent antioxidant activities against the release of ROS from the pro-inflammatory triggered neutrophils. UB significantly affected several inflammatory biomarkers that are associated with cardiovascular events, by inhibiting the production of interleukin 8 and metalloproteinase-9 , and preventing the shedding of selectin CD62L triggered by the pro-inflammatory factor cytochalasin A/formyl-met-leu-phenylalanine . UC , on the other hand, inhibited the release of elastase, a pro-inflammatory mediator responsible for extracellular matrix degradation, from the f-MLP-stimulated neutrophils. In addition to recruitment of neutrophils, monocyte adhesion to endothelial cells represents another key event in inflammatory responses. A mixture of UA and UB restricted the adhesion of Tamm-Horsfall protein-1 monocytes to the human umbilical vein endothelial cells. UA glucuronide , but not UA, UB or UB glucuronide, inhibited monocyte adhesion to TNFα-stimulated human aortic endothelial cells ; a dosage-dependent inhibition of TNFα-induced migration of endothelial cells was also shown for the above-mentioned urolithins. NO plays multifaceted roles in combatting cardiovascular diseases. Although UA, UB, and UB glucuronide did not show any effect individually at 15 µM on NO bio-availability, a mixture of the three urolithins at equal concentrations activated the expression of eNOS after a 5-min incubation and increased NO production in primary HAECs after a 24-h incubation. Overall, these in vitro studies with neutrophils, monocytes, and NO suggested the potential anti-inflammatory and cardiovascular-protective functions of urolithins. Based on the urinary excretion of urolithins by healthy volunteers after ingesting ET-rich foods or fruit extracts, three urolithin metabolic types have been defined, including A , B , and 0. Interestingly, the population of the human gut bacteria Gordonibacter urolithinfaciens correlated positively with the in vivo production of UA, but inversely with that of UB and isoUA. A recent study observed an interlinked relationship among gut dysbiosis , ET metabolism, and obesity. A relatively high percentage of metabotype B was found in the overweight-obese group, while metabotype A had a higher presentation in the normo weight than the overweight-obese group. In addition, G. urolithinfaciens levels were higher in the metabotype A than the metabotype B individuals. Further investigations should provide a mechanistic understanding of how consuming the polyphenol precursors of urolithins, in the presence of UA-producing bacteria, may reduce the risks of diseases associated with obesity. Large inter-individual variations in the cardiovascular risk biomarkers were observed in healthy overweight-obese individuals after consuming pomegranate supplement. However, after clustering the different urolithin metabotypes in these individuals, improved blood lipid profiles were evident in the metabotype B group, in a dose-dependent fashion, while there was no significant effect in the metabotype A group. Interestingly, several metabotype 0 individuals shifted to metabotype A or B after consuming pomegranate extracts. Together with the study by Selma et al. , these results suggest that consumption of pomegranate extracts may have personalized effects that are associated with the gut micro-biota and the urolithin metabotypes of the individuals. Of the various pomegranate phenolic metabolites exhibiting anti-Alzheimer activities in in vitro assays, only urolithins, including UA, UB, mUA, and methyl-urolithin B , were predicted to be capable of crossing the blood-brain barrier. Methylation of urolithins by the mammalian enzymes may further improve their lipophilicity and facilitate the penetration of the blood-brain barrier.Corroborating with the computational predictions, urolithins reduced the production of the neurotoxic, fibrillogenic β-amyloid peptide in vitro. Significant improvement of the survival/mobility of Caenorhabditis eleganswas also achieved by mUB treatment. However, empirical evidence is still needed to verify the roles of urolithins in preventing Alzheimer’s disease in vivo. Advanced protein glycation products have been implicated in the development of chronic diseases, such as diabetes and Alzheimer’s disease. The inhibitory effects of pomegranate fruit extract, punicalagin, EA, gallic acid , UA, and UB toward the early, middle, and late stage of protein glycation were compared using in vitro assays. Although all phenolic metabolites showed anti-glycation activities, pomegranate extract, punicalagin, and EA were more effective than GA, UA, and UB in preventing the glycation of proteins by fructose. The mechanistic basis underlying UA’s anti-aging activities was explored recently using C. elegans, mammalian cells, and rodent models. Feeding C. elegans with UA led to a dosage-dependent extension of the worm’s lifespan. It was shown that UA could activate mitophagy and eliminate damaged mitochondria in C. elegans, mammalian cells derived from muscle or intestinal tissues, and muscles of young and old rodents. UA was also found to be bio-available in the skeletal muscle and could improve muscle function in mice and rats. The role of urolithins in enhancing mitochondrial function and muscle quality presents promising dietary leads for improved mobility in the elderly population. Pomegranate fruits, leaves, flowers, and seeds have all been used in traditional herbal medicine for treating various illness. However, bio-active metabolites present in pomegranate fruit peel and juice have received considerable more attention than those found in other tissues. Bio-activities of the phytochemicals accumulating in tissues other than fruit should be investigated more rigorously in the future. In addition, detailed metabolite profiling and characterization of different pomegranate cultivars, grown under non-standard conditions , can be carried out to explore further the phytochemical diversity in pomegranate.

Our central claim is that the causal status effect derives from the supporting role that causal information plays in explanation. Although this idea is a long-standing theme in the study of categorization , relatively little research has directly examined how explanatory goodness relates to categorization. In this paper, we study the relationship between explanatory goodness and categorization. We demonstrate that the magnitude of the causal status effect depends on the quality of the explanation in which the causal information is embedded. We first briefly review the causal status hypothesis and present arguments that the causal status effect depends on explanatory goodness. Then we present a study to support this claim. Finally, we discuss the implications of this study for categorization more broadly. Intuitively,plastic flower buckets wholesale some components of a concept do indeed appear more important for categorization judgments. As Medin and Shoben point out, it is much easier to imagine a robin that is not red than it is to imagine a robin that lacks the appropriate genetic structure of robins: imagining a robin with, say, zebra DNA, would make many other typical properties of robins implausible .

There is a long history of research on categorization demonstrating that some features associated with category members are particularly important for categorization judgments . One prominent perspective on centrality is based on the theory- or knowledge-based view of conceptual structure . According to this framework, knowledge of a category is, in important ways, like a scientific theory, comprising a “host of mental explanations” . In this view, category knowledge is seen, not in terms of a prototypical member or collection of exemplars, but rather in terms of “an explanatory principle common to category members . Accordingly, categorization is typically viewed as an inference to the best explanation. Murphy and Medin illustrate this idea with a well-known example: a man at a party who jumps into a pool fully clothed in a business suit would probably be classified as intoxicated, not because he is similar to the prototype or to instances of a “drunken behavior” category, but because being intoxicated is the best explanation for his behavior . Despite the importance of the theory-based view for orienting research on categorization and centrality, the framework leaves unspecified the specific constraints related to background knowledge that influence classification decisions.

One important response to this challenge is the idea that information that participates in causal relations is of greatest centrality . In Ahn’s causal status hypothesis, causes are weighted more heavily in categorization decisions than are the corresponding effects. For example, in the simple case of a single causal relationship between two features, such as having wings and flying , having wings would be given greater weight than flying, and would more greatly influence categorization decisions. Why should causes play a more important role than the corresponding effects in the underlying principles of a category? According to Ahn, et al.,causal properties may be seen as generating other features: e.g., DNA produces external features such as hair color. Features such as DNA may thus be regarded psychologically as most defining or diagnostic of category membership, because they form part of the essence or core of a concept. It follows, Ahn, et al. claim, that possessing the most central features would provide better evidence for category membership than more superficial features. While not disputing the empirical validity of the causal status effect, and the critical role of causal information for many if not most knowledge-dependent categorization decisions, we worry that the exclusive focus on causal knowledge risks obscuring the supporting role that causal information plays in explanatory processes.As Lombrozo notes, explanations can’t be reduced to just the supporting causal information, as explanations entail a set of factors that go beyond the causal information per se .

For one thing, there are many kinds of causal structures —e.g., causal chains versus feedback systems—that draw on different kinds of domain knowledge. An additional factor is the preference for simple or parsimonious causal explanations over those that would invoke more assumptions . In a recent study that illuminates the relationship between the causal status effect and explanatory structure, Lombrozo has shown that the strength of the causal status effect depends partly on the type of explanation— mechanistic or teleological —in which the causal knowledge is embedded. In one experiment, participants were presented with a novel category characterized by two features and were told that one feature causes the other . Participants were then asked to explain why the animal has the second feature , and to estimate the probability that an object missing one of the features was a member of the category. Participants who provided a mechanistic explanation showed a larger causal status effect than those who provided a teleological explanation . In addition to the type of explanation, the causal status effect may also depend on the quality of an explanation. We suggest that some of the puzzling results obtained in studies of the causal status hypothesis might be best explained in terms of explanatory goodness. In one such study , Ahn, Kim, Lassaline, and Dennis gave participants a standard and two alternatives. Participants were instructed to choose which of the two alternatives should be categorized with the standard. One standard read: “This object has a high-intensity light bulb and a pouch that can contain liquid because it was designed to kill bugs.” The first alternative shared the cause, but not the effects ; the other alternative shared the effects, but not the cause . Ahn, et al. predicted that people would prefer to match an alternative to the standard on the basis of the shared cause, rather than the shared effects. Although the overall results confirmed the prediction, inspection of the results reveals that only half the items conformed to the prediction. Table 1 shows the materials used in the study and, for each item, the percentage of choices for the matching cause. While there is a noticeable causal status effect for three of the items,black flower buckets two items exhibit chance responding, and one even suggests a preference for the shared effect alternative. We suggest that the item variability can best be understood in terms of the explanatory role of the causal property being asserted for each item. One hallmark of a good functional explanation is that the intended function is causally connected to the effects to be explained. Ideally, the facts can then be understood as subsumed under a general causal law . In addition, good explanations exhibit breadth—the extent to which an explanation accounts for most if not all of the available facts ; and depth—the extent to which the local explanation fits within a larger explanatory framework, and can itself be explained . These considerations can shed light on why some items in Ahn et al.’s study conformed to their prediction, while others did not. Consider the item with the strongest causal status effect, the first item . In this case, a clear, general causal relationship links the intention with the effects: it’s generally known that bugs are attracted to light, and that bug-killing agents typically take the form of liquid pesticides. Second, the explanation is broad and deep .

Now, consider the worst-performing item: “This painting has four pillars and is red because the painter intended to draw a dog.” The causal relation between four pillars and the intention to draw a dog is extremely weak, and there is no clear explanatory framework that would link the intention to paint a dog with any of the facts. Of course, these are after-the-fact suggestions. To make this account plausible, what is needed is a manipulation of explanatory goodness. In the current study, we test the hypothesis that the magnitude of the causal status effect depends on explanatory quality: we predict that the better the explanation in which causal information is embedded, the stronger the causal status effect. The design was a 2 × 2 mixed factorial with explanatory quality , manipulated within-subjects and task order,manipulated as a between-subjects counterbalancing variable.The materials consisted of 24 stimulus sets, each based on a standard “fact-explanation” pair, which comprised a set of facts joined with a potential explanation for those facts. In the explanation-rating task the facts were presented as short sentences describing a particular situation, which participants were instructed to assume as true; and the explanation was presented as a separate sentence describing a potential explanatory account of those facts. For example, one set of facts read: “This object has a high-intensity light bulb and a pouch that can contain liquid”; and the associated explanation read: “This object was designed to kill bugs.” For the categorization task, the facts and explanation were combined into a single sentence, following Ahn, et al. . Of the 24 standards, 16 were test items, and 8 were fillers. The fillers were constructed to minimize the likelihood of demand characteristics in the categorization task . Of the 16 test items, 6 standards were taken without modification from the materials used in the Ahn, et al. study described above. In addition to the six standards from Ahn et al.’s study, we constructed an additional ten items, five of which were designed to express good explanations; and five to express poor explanations. The good-explanation items were designed so that the explanation specified a general and plausible causal relation that accounted for all of the stated facts . . These additional ten standards were adapted from materials used in studies of category-based inference by Sloman and by Patalano, Chin-Parker, and Ross . The experiment consisted of two paper-and-pencil tasks, counterbalanced for order across participants: evaluating explanatory quality and carrying out a two-alternative forced-choice categorization task. For the explanation rating task, participants were given all 24 standards, each arranged on a separate page of the test booklet. The facts were presented at the top of each page in a separate sentence, with the candidate explanation presented immediately below the facts. Participants were instructed to read the facts and the explanation, and then to provide a rating of how good or satisfying they found the explanation to be on a scale from 1 to 7 . They were then instructed to briefly explain their response. The categorization task was identical to that of Experiment 4 in Ahn, et al. as described earlier.Specifically, it was a forced-choice task between two alternatives, in which participants were instructed to choose the one alternative that should be categorized with the target. Each triad—a standard together with both alternatives—was presented on a separate page of the test booklet, with the standard at the top of the page, and the alternatives below it. For the test items, one alternative shared the cause with the standard, but not the effects; the other alternative shared the effects, but not the cause. Sample items are presented in Table 2.The eight filler items were constructed to vary the pattern so as to minimize the possibility of task demands. To this end, for four fillers, the alternatives both shared the cause with the standard, but differed on the effects; and for the remaining four fillers, both alternatives shared the effects, but differed on the cause. For each participant, item order was randomized in both tasks; for the forced choice task, left-right presentation of alternatives was also randomized.Participants completed the tasks at their own pace: the categorization task required roughly 15 minutes to complete; the explanation-rating task, 25 minutes. The results of this study provide support for the main claim of this paper: that the causal status effect derives from the role that causal information plays in an explanation; and specifically, that the magnitude of the causal status effect depends on the goodness of the supporting explanation. This finding adds support to the position that explanatory structure is important in categorization . According to this explanation-based perspective, categorization is treated as an inference to the best explanation. This perspective suggests that a primary research focus should be on investigating properties that characterize the quality of explanations, and the role that those properties play in cognitive tasks, such as categorization. A possible objection to our study would be that we simply constructed explanations that were inconsistent with participants’ prior knowledge. Ahn et al., found that the causal status effect can be eliminated if the stated causal relationship contradicts background knowledge.

What are the axes of this olfactory space? Or, in other words, do odors associated with certain parts of this space have different perceptual or physicochemical properties? Previous studies found axes that correlated with perceptual odor pleasantnes and physicochemical properties such as molecular boiling point and acidity . We checked for associations with all of these properties, and supporting previous findings found that points corresponding to pleasant and unpleasant odors occupied different parts of the space. We note that this analysis of pleasantness rankings was based only on odor components from tomato and strawberry data sets, for which these rankings were available. Thus, the pleasant-unpleasant odor axis can be identified even solely using fruit odor components. The direction most associated with a change in pleasantness value is marked by the red line in Fig. 4 . For odor mixtures produced by individual fruit samples, we use the “overall liking” rating assigned by humans to fruit samples as a measure of pleasantness . To test how well the identified pleasantness axes can predict measured pleasantness rankings for novel samples,procona valencia buckets we regenerated this axis using only strawberry samples and use it to predict pleasantness ranking for tomato samples. The correlation was significant, with correlation coefficient R = 0.34 and P = 0.01 .

The pleasant- ness values could also be assigned to individual odor components based on the correlation between the odor concentration in a mixture and mixture pleasantness computed over samples . This measure of pleasantness produced an even stronger correlation between pleasantness and odor coordinates within the space,R = 0.66 and P = 3×10−7 . We also computed these correlation values using different odor components from those used to generate the pleasant-unpleasant axis for odor components in Fig. 4C. Specifically, we used two-thirds of randomly selected odor components to generate this axis; we computed the correlation value using the remaining components. The pleasantness axis for odor components had a similar orientation to the one for mixtures . In addition to the pleasantness axis, we could also find axes that were strongly associated with two other properties: molecular boiling point—which is probably a reflection of volatility—and acidity, both of which showed significant correlations . We assigned acidity for individual odors as the correlation coefficient be- tween its concentration and fruit sample acidity measurement. Be- cause the space is essentially 2D, the three axes of odor pleasantness, acidity, and molecular boiling point are not independent. In other words, knowing the coordinates along the molecular boiling point and acidity axes, one can predict the position along the pleasantness axis. That is, the identified mapping to a sphere in a hyperbolic space makes it possible to predict, with correlation R = 0.34 for natural mixtures and with R = 0.66 for monomolecular odors, how perceptually pleasant these odors are based on their projections on the acidity and volatility axes.

The observation that the odor mixtures can be mapped onto a continuous metric space is consistent with a previous vector-based model of human olfactory perception . This model posits that the perception of odor mixtures is based on a combination of the mixture components or, in other words, that there is an underlying set of coordinates that can represent olfactory mixtures. Previous analysis of the Dravnieks database containing human perceptual descriptions of > 120 monomolecular odors showed that the perceptual space is likely to be curved. Qualitatively, the points were found to form a “potato-chip” surface . This can be a signature of the hyperbolic space; potato-chip or saddle-like surface have a negative curvature and serve as an everyday example of hyperboloid surfaces. To quantitatively test whether the perceptual space is described by hyperbolic geometry, we applied the Betti curve method to the Dravnieks database. First, we found that Euclidean spaces were not consistent with measured Betti curves . The first Betti curve could not be matched to the data in terms of its area for any dimensionality of the Euclidean space . Second, we found that the full hyperbolic space of varying dimensions could match the area of the first Betti curve. However, only hyperbolic spaces with small number of dimensions could also simultaneously match the area of the second Betti curve . The 3D hyperbolic space produced the best fit, with larger dimensions yielding increasing deviations. Hyperbolic spaces with dimensions nine and above could be excluded with P < 0.034. The third Betti curve was essentially zero and is not shown here.

One may notice that the first and second Betti curves were not as regular as in the case of odorants and contained multiple peaks. It turns out that the biphasic nature of the Betti curve could be explained by the nonuniform distribution of points across the two angles . Unlike in the case of olfactory stimulus spaces that are sampled approximately uniformly, here, the distribution of points obtained using MDS is not uniform and clusters in one-half of the space. Sampling points from this embedding yields biphasic Betti curves that match those derived from perceptual data . Specifically, P values for L1 differences between Betti curves derived from data and MDS fits were P = 0.32 , P = 0.20 , P = 0 , and P = 0.06 . The MDS distances also better correlated with perceptual distances when we carried out MDS in the hyperbolic space compared to Euclidean space . Our results highlight the importance of hyperbolic curved geometry for understanding how natural odors are represented in the nervous system. Overall, we find that both the statistics of natural odor mixtures and human odor perception can be mapped onto hyperbolic spaces. In the natural environment, hierarchical biochemical networks produce odor components. Hierarchical networks can often be approximated by trees and, therefore, by hyperbolic spaces . We find that most natural odor components fall near the boundary of the observed hyperbolic space, corresponding to leaves of the trees . At the perceptual level, we also found hyperbolic organization. However, in this case, the odors selected for the Dravnieks database did not sample the human perceptual space uniformly . Hyperbolic perceptual organization is likely to be general across different sensory modalities. There are two reasons for this. First, neural networks that give rise to perception are hierarchically organized, and as we have seen in Fig. 1, this can lead to hyperbolic geometry. Second, individual neurons have limited response ranges. Because of response saturation, small changes in neural responses near their limit correspond to exponentially large changes in the input values. This compressive mapping is similar to the Poincare disk representation of the hyperbolic space. There is evidence that visual, haptic,procona buckets and auditory perceptual spaces are all hyperbolic . Adding olfactory perception to this list could help explain why humans can map odors to auditory pitch and to colors. Noteworthy is the low dimensionality of both the physical odor space and perceptual odor spaces. In both cases, the curved space contains approximately three dimensionsdespite the fact that the data vary in > 50 dimensions associated with different samples of natural odor mixtures and according > 100 perceptual descriptors. The low dimensionality of the environmental odor space could be a general property of natural odors because it occurred for odors as diverse as fruit and mammalian urine odors. Note that all four natural odor data sets were described by the same 3D hyperbolic space with exactly the same radius . This property could make it easier to represent data from different data sets within the same space. For example, odors from strawberry and tomato could be represented jointly within a single 3D space .

We could not combine data from other data sets because, for example, there were no overlapping components between fruit odors and mouse urine data sets. It is possible that representing all possible natural odors will increase the dimensionality of the overall space. Another possibility is that introducing odors from different sources will “fill in” the inner part of the hyperbolic space. The natural odors considered here mapped onto a surface of a hyperbolic space. Odors produced by biochemical pathways of different complexity are likely to map to surfaces with a different radius, filling in the space. This possibility is especially interesting because it would provide a link to the filled 3D hyperbolic space that we find for perceptual data, which was obtained using diverse classes of odors. At the same time, the perceptual odor mapping reveals that odors tested so far concentrate on one side of the space , whereas natural odor components cover their respective space rather uniformly . These analyses thus suggest perceptual coordinates that are yet to be explored. The match in dimensionality between the environmental and perceptual spaces would not have been expected a priori. The matching dimensionality between the input and perceptual spaces can help avoid nonlinear distortions that would necessarily arise when mapping two nonlinear spaces of different dimensionality. These distortions are known to exist in vision where we perceive distances in a compressed way: The moon appears disproportionately closer to us than would be based on the actual Euclidean distance . We also plot equidistant and parallel lines differently, which is one of the key signatures of the hyperbolic space. Similar distortions arise in the haptic space . The matching geometry between the input and perceptual spaces in olfaction may therefore serve to minimize these distortions in odor perception. Overall, the ability of the perceptual system to resolve points in the low-dimensional odor space would depend on the number and tuning properties of sensory receptors . We followed procedures from to generate Betti curves for samples taken from spaces with different geometries. This renders the algorithm’s results invariant under monotonic transformation of values, for example, due to nonlinearities introduced at the measurement stage. However, this property can also be used to assign a distance between points based on the correlation in the activity of two units in a network or, as in our case, between two odors across different samples. All monotonic functions will yield the same result. We chose Di j = −|Ci j|, where Di j is the assigned distance between odors, and |Ci j| is the absolute value of the correlation coefficient of odor concentrations among a set of points. This definition ensures that stronger correlations corresponded to tighter connections and smaller geometric distances, as in. The first three Betti curves turn out to be quite sensitive measures of the distance matrices and can be used to find underlying geometries consistent with the data. In addition to random spaces, we screened two kinds of geometric structures: Euclidean spaces of different dimensionality and hyperbolic space [we used the hyperbolic ball model with curvature z = 1] with different parameters. In each space, we uniformly sampled points based on the metric of the space. In a d-dimensional Euclidean space, the points were uniformly distributed in a d-cube with Euclidean distance. For a d-dimensional hyperbolic ball model, we used partial space by setting the minimal radius Rmin and maximal radius Rmax for the ball. This choice of the model was motivated by the fact that hyperbolic space approximates hierarchical tree-like networks, with odors reflecting leaves—the neighborhood of the surface. We use the differences between descriptions across odors, because in this case the absolute value of the descriptor matter, unlike in the case of odors where correlations were a more appropriate measure. When fitting the data using geometric models, no noise was added to distances in models. We also tested the sensitivity of the Betti curves to noise in pairwise perceptual distances between odors. This was done by computing perceptual distances based on randomly selected subset of 120 out of the total 146 descriptors. The variability in the resultant distance values was proportional to the mean distance . Importantly, the relative error in the integrated Betti values across these samples was the same as the relative error of the distances themselves . Thus, although the Betti curve construction evaluates data structure globally, it is not driven by variability in larger distances. In the case of the perceptual dataset, we found that the full hyperbolic space better described the data rather than a shell, and therefore the minimal radius was set to zero. We optimized maximal radius of the hyperbolic model, which is a measure of its curvature, to fit the integrated Betti value of the first Betti curve.

Lack of nutrition education for students was the third most commonly cited barrier to successfully providing additional servings of fresh fruit. Although it was intended that the program include a nutrition education component, schools were provided with only 1 cent per meal served for all nonfood expenses related to the program, including nutrition education and promotion. Yet, in spite of the limited funding, over half of the schools did report using some nutrition education or promotional materials.Although slightly more than half of nutrition directors had implemented some form of staff training, 40% reported that staff needed more training in the areas of fruit and vegetable handling and management and nutrition education in order to more effectively market fruits and vegetables to students. Nutrition, Family and Consumer Science advisors are poised to provide this kind of education and training.The 10 cents per meal the program provided was substantially less than the amount required to cover a supplementary serving of fruit. Without considering the labor and other costs associated with serving additional fruit,plastic planters wholesale the cost of the fruit alone was 3 cents higher than the 10 cent reimbursement, 4 cents above the 9 cents designated for food cost.

The cost per serving for apples, oranges and bananas is 12 to 15 cents, whereas the other fruits offered in the program range from 14 to 25 cents per serving . The 1 cent intended for all other costs, including labor, education and administrative expenses, was totally inadequate; labor costs for school food service departments usually account for 30% to 50% of expenses, and costs associated with storage and produce loss from perishability add to the expense of handling fresh produce. If the California Fresh Start Program were to offer a greater variety of fruits and higher-quality fruit, as students would prefer, fruit consumption would likely increase, leading to substantial increases in labor costs and costs of facility modification, as well as produce costs. Most nutrition directors reported that the program reimbursement was inadequate to cover the cost of implementing the program. Those that did report nonfood expenses spent an average of $2,784, primarily on small equipment and educational materials. Nutrition directors identified cost as the main barrier to providing a greater variety of fruits. They tended to serve fruit whole to reduce waste, increase shelf life and reduce labor costs associated with chopping fruit. They served apples, bananas and oranges most frequently because they are the most affordable. Student favorites such as strawberries cost much more . To keep costs down, nutrition directors obtain their fruit whenever possible through the commodity food and Department of Defense programs.

According to the nutrition directors, they could serve a greater variety of produce if these programs offered a more consistent supply and greater variety of fruits and vegetables. Despite the perceived inadequacy of the reimbursement, many nutrition directors thought the program was helpful financially and made it easier for them to provide more fruits because enrollment in the breakfast program increased. Higher participation rates in school breakfast might help to improve the bottom line by bringing in more federal reimbursement dollars. Participation increased slightly during the program, but the increase was too low to be statistically significant; it is not clear what would have happened over a longer period of time. During the program, more California grown fruit was sold and distributed because the quantities of fruit and varieties of fruit purchased by schools increased. Using production records from schools participating in the program evaluation and interviews with selected produce distributors, we were able to estimate the proportion of produce the schools purchased that was California grown .If all schools in California were to increase fresh produce offerings at breakfast, annual school purchases of California-grown fruit would increase by an estimated 26 million servings, valued currently at approximately $4 million per year. These estimates are based on an average increase of at least one-third of a serving of fresh fruit per school breakfast meal served, an assumption that 47% of the fresh fruit served would be California grown, and an average cost per serving of 15 cents. Given this scenario, an additional $8.3 million would be spent on fresh produce, of which approximately $3.9 million would be spent on California grown produce and $4.4 million would be spent on fresh produce from other sources.

If students’ favorite fruits, which are primarily California grown, were served more often, our findings suggest students would take even more fruit at breakfast and therefore the increase in the value of school purchases of California grown produce would be even higher. According to school nutrition directors, produce vendors were a program asset, making it possible to increase fresh fruit servings by providing dependable service, high quality and reasonable prices, and providing access to locally grown produce. While nutrition directors were enthusiastic about using more locally grown produce, some felt that they needed a go-between to procure the produce from local growers. Some reported that there were few, if any, local growers, and even if there were multiple local growers, it would be difficult to work directly with them. Directors expressed concern about supply, distribution, dependability, food safety and cost issues when dealing directly with local growers. Produce vendors were seen as necessary intermediaries between the schools and the growers as some directors felt that food service departments are not equipped to deal with many small growers. They preferred to continue dealing with the major suppliers with whom they have ongoing relationships. Increased purchase of California grown produce can be a win-win for schools, students, distributors and farmers. Food distributors indicate that they prefer to purchase fresh fruit from California farmers, when available, as it is more affordable than fruit imported from out of state. Schools benefit from these savings, and farmers benefit from an increased market demand. The ultimate beneficiary is the student, whose increased consumption of fruit will contribute to long-term health. While the California Fresh Start Program was designed to provide more fruits and vegetables to students, its application to the breakfast program led to an overwhelming emphasis on fruit, and it is therefore impossible to draw conclusions about its potential impact on vegetable consumption from our data. However, other studies have indicated that increasing student vegetable consumption at other times in the school day presents greater challenges than are found with fruit . A systematic review of 27 school-based programs designed to increase fruit and vegetable intake found that although the programs moderately improved fruit intake, they had minimal impact on vegetable consumption. The authors called for additional studies to address barriers to changing dietary behavior, particularly in relation to vegetables . As implemented,plastic plant pot the California program was effective in increasing the amount of fruit, particularly the amount of fresh fruit, offered to and taken by California school children each day. Further, the variety of fruits offered, especially those that were fresh, increased substantially. The success of the program demonstrated that schools can have a positive impact on students’ consumption of fruit, which is particularly important since produce consumption at school is lower than at home. Piloting the program also provided lessons for the future implementation and expansion of such a program. Given the well-documented health risks that poor nutrition poses for California’s school children and, at the same time, the likelihood that a school fruit program may decrease children’s intake of unhealthy snacks at school , it is critical to closely examine those lessons. The significant increase in the number of fruit servings students took at breakfast during the program was observed even in the absence of adequate funding to promote the effort or to upgrade facilities and equipment so that the fruit could be served in a way that would make it more attractive to students. Infusing additional resources for training, technical support and facilities upgrades, including, for example, improvements in storage capacity, adequacy and attractiveness of cafeteria seating, and creative presentation such as the use of salad-type fruit bars and point of-service displays, could lead to even more substantial increases in the servings students take.

The program also was successful at shifting student consumption away from fruit juice toward fresh fruit. When schools serve less juice at breakfast, students take more fresh produce. Fruits are a healthier option than juice because of their higher levels of fiber and associated micronutrients. While the program resulted in a doubling of fruits offered to students at breakfast and a doubling of fresh fruit taken, a limitation of the study is the lack of assessment of the amount of fruit consumed. While not assessed in this study, improved variety and appeal in produce offerings, improved facilities, and more nutrition education could all potentially result in higher total consumption as well. Our evaluation of the program suggested that school food service personnel faced a dilemma: If they took steps to improve the variety, presentation and promotion of fruit, their labor and food costs would increase beyond the 10 cents per meal provided by the program. At the same time, more students would likely take more fruit servings, particularly servings of fresh fruit, thereby further straining food service budgets. Our study suggests that additional financial resources will be required to ensure that most or all students take the recommended two servings of fruits and vegetables at breakfast. If the program reimbursement were increased to 15 cents to better cover estimated actual costs and if all eligible schools participated, the total reimbursement figure would be about $26 million per year. School programs would further benefit from additional funding to make food service facilities adequate to store, prepare and serve fruits and vegetables in a safe and appealing manner. Interestingly, the USDA’s Food and Nutrition Service recently estimated that the final meal pattern ruling mandating increases in whole grain and fruits in the School Breakfast Program would necessarily increase the cost of food and labor by 14 cents, an amount similar to our estimate for fruit . As part of the Healthy, Hunger-Free Kids Act of 2010, an extra 6 cents per meal in reimbursement was provided for school nutrition programs that complied with the mandate to increase the kinds and amounts of fruits and vegetables. As the final provisions of this act are implemented in schools across the nation, it will be important to evaluate school programs. California has recognized the need to reverse the trend toward poor youth diets and has acknowledged the responsibility of schools to promote health. Unfortunately, lack of financial resources led the state to discontinue funding for the California Fresh Start Program after the pilot. However, new programs can benefit from lessons learned from the California program. There are currently changes taking place in school nutrition policy at the federal level. Partnerships among influential organizations and sectors, including growers, schools, public health agencies and others have been suggested as a lynchpin of the National Action Plan of the National Fruit and Vegetable Alliance . The health of future generations depends upon our commitment to ensuring that everything possible is done to help today’s youth adopt healthy food habits.Aging can be modulated by genes and lifestyle. For instance, specific gene variants of insulin-like growth factor-1 receptor and fork head box O3A are associated with longer lifespan in centenarians. In terms of lifestyle, one of the most studied interventions that delay aging is caloric restriction , which can increase lifespan in organisms ranging from yeasts to primates. Diet composition also influences the aging process, with low protein diets and high phytochemical intake being associated with a longer lifespan. Notably, a recent analysis suggests that the heritability of human longevity is below 10% , indicating that lifestyle choices play a major role in influencing aging and longevity. Since interventions such as CR and dieting are difficult to implement and maintain over a long period, interest has focused on identifying molecules that produce effects similar to CR . This endeavor is based on the observation that signaling pathways that are modulated by CR, including 5’ adenosine-monophosphate-activated protein kinase , mammalian target of rapamycin and sirtuin-1, can be targeted by small organic compound. Activation of these pathways induces autophagy, mitochondrial bio-genesis and expression of antioxidant and detoxifying enzymes, which together can improve cellular function. In a manner similar to CR, several organic compounds labeled as CR mimetics promote physiological functions and reduce the development of chronic diseases, thus improving both health and longevity.

Viable and non-viable seeds in yellow starthistle seed heads are easily distinguishable based on color and shape. Because yellow starthistle requires pollination to produce viable seeds , non-viable seeds represent pollen limitation occurring during the 4-hour period that the flowers were exposed to pollinators. All seeds were counted to compare ratios of viable to non-viable seeds. Any seed predation was noted, and when possible, the seed predator was identified.All analyses were done in R 2.15.1 . Because each site had an AM, Mid-Day, and PM observation event, there were a total of 36 observation events, each with unique wind and temperature recordings, and visit observations of the 15 bee morphotypes. From these, we calculated the total number of bee visitors, total number of bee morphotypes, Shannon diversity of morphotypes, and morphotype evenness. Shannon diversity and evenness were calculated using the R package vegan. The spatial autocorrelation of all bee visitor response variables was assessed by Mantel tests in R package ade4,plastic pots large using the average values for each time of day at each site. Spatial autocorrelation was not detected .

To test for the effect of land use type on each of the response variables we used a generalized linear mixed model using the R package lme4. We designated land use type, bloom category of flowering patch, observation time period, wind, and temperature as fixed effects and site as a random effect. Natural land use and AM observation time period were the model baselines for the categorical variables of land use type and observation time. Shannon diversity and evenness were fit with Gaussian distributions while all other variables were fit with Poisson distributions. In comparing the ratios of viable seeds to total seeds vs. the ratio of viable seeds to counted stigmas, we found that there was a strong correlation between these metrics. To look at the effect of land use type on seed-set, we therefore decided to utilize the ratio of viable seeds to total seeds in each seed head that did not experience seed predation, because of error in counting the number of stigmas . We then used a generalized linear mixed model fit with a Binomial distribution, with land use type as a fixed effect and site as a random effect. Finally, we tested for an effect of floral visitor observations on yellow starthistle seed set at each site. We averaged the number of visits from each morphotype across temporal observation events at the same site. Morphotypes that averaged at least one visit per 30 minute observation window were included as fixed effects in alinear mixed model fit with a binomial distribution, with site as a random effect and the ratio of viable to total seeds as the response variable. We also modeled the effects of total bee visitation, morphotype richness, and morphotype diversity on seed set ratios.

Our results show that rates of bee visitation and seed set vary among urban, agricultural, and natural landscapes, demonstrating the importance of land use in the dynamics of plant-pollinator interactions. We suggest that these effects are at least in part explained by floral availability, a vital bee resource, which can be highly variable among different land use types. For example, in August there are few plants in flower besides yellow starthistle in the natural areas of Contra Costa County, California, whereas in urban and agricultural areas there are many exotic plants and supplementary inputs available . From pantrapping of bee specimens in the region , we know that total bee abundance is highest in the spring in natural areas. However, towards the end of the summer when yellow starthistle is in flower, there is little difference in collected bee abundance between human-altered landscapes and natural areas, and human-altered areas may even exhibit overall higher bee abundance. Our results of bee visitation to yellow starthistle support this pattern. Agricultural areas have large populations of managed honey bee colonies, so one would predict visitation to yellow starthistle by honey bees to be positively associated with surrounding agricultural land use. By contrast for native bees , the highest rates of visitation to yellow starthistle were in sites with more surrounding urban land use.

Urban gardens have many exotic plants, often selected for aesthetic purposes, many of which are in flower later in the season than most California native plants. In addition, many of the plants in urban areas both directly and indirectly receive supplementary resources, particularly water, that further extend their flowering time. Even though agricultural areas also have supplementary resources, the main crop in flower in East Contra Costa County later in the season is maize, which is wind pollinated. There may be multiple impacts of exotic plants in urban areas. By filling the phenological flowering gap noted above, they may help attract even larger populations of bees into the urban landscape. In addition, bees in urban sites may be behaviorally more likely to visit non-native plants due to the increased encounters they have with novel plants. In agricultural and natural landscapes, a positive correlation between pollinator visitation and seed set is typical. Surprisingly in our system, in human-altered landscapes, higher total observed bee visitation did not result in higher proportions of seed set, as would be expected. In fact, urban areas, despite receiving the highest rates of native bee visitation, exhibited the lowest rates of seed set. Conversely, natural areas, which received the lowest amount of total bee visitation, had the highest rates of seed set. We suggest 2 possible explanations for this discrepancy between pollinator visitation and rates of seed set: 1) pollinator efficiency; and/or 2) the composition of the local flowering community. Depending on the plant, certain pollinator species are much more effective than other. For example, Osmia, Habropoda, and Apis, have been found to produce varying amounts of seed set as a result of a single visit to blueberry, but these results vary slightly depending on the blueberry variety. In the case of yellow starthistle, it is likely that the most frequent visitors are perhaps not the most efficient. When we directly compared average seed set at each site against visitation rates, we found a significant positive association with the medium hairy leg bees. The medium hairy leg bee morphotype includes those species which fall in both the Tribes Emphorini and Eucerini. Emphorini are known to largely be oligolectic , meaning they specialize on certain plant groups, which theory suggests would make them more efficient pollinators than generalists. The medium hairy leg bee morphotype was not significantly associated with any of the land use typesIt was also the only group that was observed most frequently during morning sampling, perhaps reflecting a difference in when yellow starthistle is most receptive to pollination.

Despite the overwhelming abundance of honey bees in agriculture areas, we did not observe higher seed set in those regions, consistent with the observation that honey bees can be poorer pollinators than other species.It is also important to note that this study used a morphotype classification,black plastic nursery pots and there may be multiple species that fit within the same morphotype that provide varying degrees of pollination service. It is possible there are rare, but highly efficient, pollinators that were rarely observed during the sampling period, or were lumped together with a more frequently observed morphotype. An alternative explanation for the lack of an association between floral visitation and seed set is that higher plant diversity in urban and agricultural areas may decrease pollinator efficiency. Previous research has shown that invasive alien plants can have a negative effect on native plant communities by acting as attractors for pollinators, or decreasing pollinator efficiency by providing a wider range of resources for pollinators to visit, with the consequence that visitors transfer pollen from non con-specifics, potentially clogging stigmas and reducing pollination success. In this case, our target plant, yellow starthistle is indeed considered an invasive alien plant, but the hypothesis of it being in a novel diverse community could lead to a similar effect on the frequency and quality of pollination services that it receives. In sites where there are many other potential plants to visit and accompanying decreased floral fidelity leading to diverse pollen loads, one predicts decreased pollinator efficiency. Abundant sources of exotic plant pollen could occur in areas where there is a greater diversity of nearby plants for pollinators to visit. This explanation might account for the observation that shield-tipped small dark bees were negatively correlated with seed set. We selected yellow starthistle as the target plant for this study because of its ubiquitous distribution, reliance on pollination, and its attraction for a wide set of visitors; it is also a highly invasive and undesirable plant. Previous research on yellow starthistle has found that its invasion can be facilitated other non-native pollinator species such as the honey bee, Apis mellifera, and the starthistle bee, Megachile apicalis, which is included in the medium striped hairy belly bee morphotype. However, the abundance of bees in both of these 2 morphotypes were most closely associated with agricultural areas, which did not have the highest rates of seed set as would be predicted by visitation alone. Our results indicate clearly that bee visitation in human-altered landscapes can be higher than that in comparable natural areas, especially towards the end of the flowering season when there are few resources available in natural landscapes.

Because the response of bee visitors to land use change depends on species specific requirements and these pollinators also have variable effects on plants, understanding the effect of land use change on pollination services requires knowledge not only of which pollinator groups shift to the human-altered landscapes, but also the rate of pollination that those groups have on the plant species in those landscapes. Future research will benefit from looking at a wider range of plants with a different range of target pollinators and that flower earlier in the year to better tease out these hypotheses. If the patterns of bee visitation and seed set that we observed are indeed consistent across other plant species, the novel plant communities created in these human-altered landscapes and the generalist bee species that are favored in such landscapes will lead to a reduction in overall pollination services.Diet and lifestyle patterns play a significant role in the development of Metabolic Syndrome and the diseases it precipitates, namely cardiovascular disease and type 2 diabetes mellitus. In particular, MetS development is associated with the consumption of foods typical of the Western diet, including red meat and fried foods. The Western diet is characterized by excess intake of energy, readily available carbohydrates and fats that result in postprandial hyperglycemia and hyperlipidemia. Three large scale prospective cohort studies support the relationship of postprandial lipemia and risk of cardiovascular disease. Postprandial inflammation and oxidative stress are postulated to be products of hyperglycemia and lipemia and are associated with impaired insulin sensitivity. Impaired insulin responses to Western meals exaggerate postprandial hyperglycemia and lipemia, creating a vicious cycle leading to vascular dysfunction and damage that, over time, likely augments risk for CVD. Reducing transient fluctuations of these postprandial responses is one lifestyle modification for the prevention and management of MetS. The addition of certain foods to the diet may attenuate these postprandial responses. Foods rich in dietary fiber have been shown to modulate postprandial lipemia and aid in glycemic control.Antioxidant-rich foods have been shown to attenuate postprandial oxidative status markers and increase plasma antioxidant capacity. Few studies have investigated the postprandial response to beans and only one has studied black beans. These few studies have demonstrated that different types of beans elicit different biological responses. This may be in part due to dosing differences, polyphenol bio-availability, and endogenous factors. For example, polyphenols modulate digestive enzymes, eliciting beneficial effects on the postprandial responses. Anthocyanidins in particular, which are found in black beans, have been shown to inhibit α–amylase, maltase, and sucrase activity, which lower postprandial blood glucose. Further, metabolic health benefits of beans are likely related to the functional effects of their inherent dietary fiber and polyphenol content, the latter being questioned as an effect of the general antioxidant properties of polyphenols or a specific effect of the polyphenol structure. Nowadays, it is popular to isolate and sell functional components of foods as dietary supplements and many supplements are marketed for their “antioxidant” properties.

At the time of this study, the trees in the control and treatment bioswales were fully established and approaching mature size. Measurements recorded the differences in surface runoff dynamics and pollutant reduction rates, as well as tree and shrub growth. This study provides new information on the long-term effectiveness of engineered bioswales in a region with a Mediterranean climate. The water collection system was installed in 2007 to collect composited samples from natural runoff . In this study, surface runoff samples from the control site were collected at a high frequency using grab samples to better observe pollutant concentration dynamics for each experiment throughout a storm hydrograph. A test run was conducted on 10 October 2013 to determine the optimal runoff sampling time intervals and the number of samples needed to capture the peak and total loadings. For the test run, the water soluble fertilizer was applied to both of the sites at a rate of 2.24 g m−2 . Grab samples were collected at a 10-min frequency over a two hour irrigation period from the control site. Water samples were collected immediately before the runoff was directed to the underground tank.

The water samples were coarse filtered during sample collection with coffee filters to remove large tree leaves,square plant pots grass clippings and large soil particles. Based on the results of the test run, the water sampling frequency for subsequent trials was extended to six hours with a variable sampling interval to better characterize the runoff pollutant pattern. The composite water sample from the treatment site was used for calculating the total loading of the treatment site where little surface runoff occurred in this study. A 5.1 cm diameter and 0.9 m long PVC drainage pipe was vertically installed into the middle of the treatment bioswale to collect a representative water sample for monitoring pollutants concentration dynamics in the bioswale. The treatment site water samples were not affected by successive flow from the control site because the treatment site was located upslope of the control site. The water sample collected from the control site was surface runoff, which was not affected by subsurface flow of the treatment site because of the site’s relatively flat surface. The pH ranged from 7.90 to 8.24 for the control site and 8.07 to 8.20 for the treatment site. The relatively high pH values indicate that alkalinity was not elevated by the ESM used in this bioswale. The high pH and alkalinity results from the relatively poor quality of the irrigation water originating from groundwater. The groundwater sources are from marine sedimentary rocks of the Coastal Range, which have high salt and carbonate levels.

The irrigation water supply had an average pH value of 8.79, and its hardness was as high as 890 ppm. Another factor that contributed to the accumulation of salts in the soil was the 2013–2014 drought, which resulted in very little soil leaching in the year prior to this study . Without leaching from winter precipitation, the salts accumulated in the surface soil layer. The cations were mainly Na+ , Ca2+, and Mg2+, and the anions were mainly Cl−, SO4 2−, and HCO3 −. Concentrations of these major cations and anions did not show significant variation from sample to sample, indicating that their dominant source is the irrigation water. The high TIC in all water samples reflects the high carbonate concentrations of the groundwater used for irrigation. On average, the bioswale with ESM reduced pollutants carried in surface runoff by more than 99.5%. The average pollutant loading reduction rate was 99.6% , 99.5% , and 99.4% for the LPL, MPL, and HPL experiments, respectively . This treatment bioswale had slightly higher pollutant reduction rates as compared to the bioswale with ESM installed adjacent to a parking lot in a previous study. The parking lot bioswale reduced the nutrients by 95.3% and organic carbon by 95.5%. The peak pollutant concentration reduction rates found in this study were a minimum of 53% higher than those reported in the parking lot study. One possible explanation for this difference is that the trees and shrubs in the bioswale were more extensive and older than the tree in the parking lot site. Tree and shrub roots can function as a biofilter, where pollutants are immobilized, transformed, or degraded.

Although data were not available for below ground biomass, a more extensive rooting system and associated microorganisms in this study’s bioswale could be partially responsible for its improved performance. Another possible explanation is the difference in pollutant inflows. The primary pollutant source in the parking lot study was from atmospheric deposition, with lower concentrations when compared to the fertilizer rates applied in this study. Storm water BMPs, such as bioswales, are reported to have higher removal rates when treating storm water with high inflow concentrations. Concentration of Zn, Cd, Ni, Cu, and Pb are key water quality concern parameters. They were excluded from this analysis because their concentrations were below detection levels in the irrigation source waters and these metals are not identified as impairments in the study area.The interpretation of results from this study is subject to some limitations. Pollutants can leave the system via infiltration deeper into the soil and potentially enter the groundwater. Deep leaching can be an important flow path affecting the fate of pollutants, and was not included in the scope of this study. Caution should be taken regarding the potential for groundwater contamination when considering the use of ESM in bioswale projects. In this experiment, the trees were eight years old and their root systems were well established. Trees received excess surface irrigation runoff during the hot/dry summer. Because the ESM in this study was 75% lava rock it may not retain enough moisture for tree roots during long dry periods. Trees in bioswales with ESM may require more irrigation than trees in native soils, especially for establishment. In this study, the pollutants were artificially added to the system by using dissolved fertilizer. Actual storm runoff includes pollutants from atmospheric deposition and has a more complex mixture of pollutants. These factors introduce uncertainty in extrapolating the pollutant reduction efficiency of the bioswale to other sites. It is unclear whether all of the pollutants retained by the bioswale were fully retained by the vegetation and soil, or if a portion of these pollutants were only temporarily immobilized in the system by the soil-tree root system. The bioswale system tested in this study was eight years old, relatively young when compared to its 20 to 30 year life expectancy. Long-term monitoring of system performance is needed to document bioswale performance over longer time periods typical of urban green infrastructure. Additional research is needed that follows the fate and transport of pollutants after infiltration. In particular, chemical analyses of soil and tree samples are needed to understand the fate and transport of the pollutants in the bioswale system. The length of lifespan of a particular normal cell of any organism is predetermined.

Similarly, the length of lifespan of all organisms is pre-determined by their genetic makeup and their external and internal environments and diet-related factors specific to an organism. Therefore, the length of lifespan can be increased or decreased by manipulating the environment, diet and genetic factors only by small extent. The length of lifespan of the organisms can also be impacted by differential rates of senescence of cells and organs that ultimately lead to the death of the organisms. The differential rates of cellular senescence are influenced by several confounding factors,plastic potting pots such as external and internal environments, diet and genetic factors. Because of these confounding factors that can impact rates of progression of degenerative changes in the organs, it is almost impossible to study aging in the absence of organ pathology. Based on numerous studies on aging in vertebrates and invertebrates, a recent informative review has suggested that oxidative stress theory of aging can only be applied to conditions in which age associated pathologies are included. Furthermore, it was suggested that in environment with minimal stress, oxidative damage plays little role in aging. This suggestion can be argued on the fact that little oxidative damage may take longer time to deregulate protective transcription factors, adaptive responses to stressors, and repair mechanisms, and thereby extending the lifespan of the organisms more than that produced by higher oxidative damage which can deregulate above biological functions in shorter time. Using vertebrate and invertebrate models, some major biochemical and genetic factors that are associated with aging processes have been identified. They include increased oxidative stress and chronic inflammation, decreased adaptive response to stressors, post-translational protein modifications, mitochondrial dysfunction, decreased of proteasome and lysosomal-mediated proteolytic activity, shortening of telomeres and transcriptional deregulation. Among these, the theory of oxidative stress is most extensively investigated in various experimental models, using pharmacological agents, antioxidants, anti-inflammatory agents and deletion of one or more antioxidant enzymes as well as of mitochondrial complexes. Depending upon the experimental models, experimental designs, and substrate used to assay oxidative stress and criteria of oxidative stress, the role of oxidative stress in aging has been substantiated or questioned. We hypothesized that increased oxidative stress may be one of the primary early events that causes chronic inflammation, transcriptional deregulation, post-translational protein modifications, mitochondrial dysfunction, decreased of proteasome and lysosomal-mediated proteolytic activity and shortening of telomeres. Invertebrate models, such as Caenorhabditis elegans has been extensively used to evaluate the role of oxidative stress in aging primarily due to shorter lifespan of about 3 days and ease of genetic manipulation. This review analyzes recent published studies on C. elegans on the role of oxidative stress in determining the length of lifespan by generating mutants that show suppression of mitochondrial function or lack of superoxide dismutase .

Caenorhabditis elegans has been extensively used to investigate the role of oxidative stress in aging by measuring the length of lifespan. Mitochondria are considered the major sites for the production Reactive oxygen species , although ROS are also produced outside the mitochondria. In order to demonstrate the impact of oxidative stress, several mutants of C. elegans were generated. They include mutations in four clock genes , mutation in the iron sulfur protein of mitochondrial complex III, mutation in the gene NUO-6 and mutation in the gene daf-2. The effects of mutations on oxidative stress and lifespan are summarized in Table 1. The clk-1 gene encodes an enzyme that is necessary for the biosynthesis of ubiquinone that is required by the mitochondria to generate energy. Mutation in the clk-1 gene increases the life span by slowing down mitochondrial activity due to reduced availability of ubiquinone. This slowing of the electron transport chain would reduce oxidative stress. The role of reduced oxidative stress in extending the lifespan is further supported by the fact that overexpression of clk-1 gene in wild-type C. elegans increased mitochondrial activity and shortened the lifespan. A mutation in the iron sulfur protein of mitochondrial complex III causes low oxygen consumption, reduced oxidative stress and increased lifespan. Mutation in the daf-2 gene which codes for a member of insulin receptor family increased lifespan and enhanced resistance to oxidative stress. In this daf-2 mutant, expression of the SOD-3 gene, which encodes mitochondrial Mn-superoxide dismutase, was much higher than in the wild type. This implies that the increased levels of SOD-3 in the daf-2 mutant reduced oxidative stress and thereby increased lifespan. Mutation in the gene NUO-6 which encodes complex I of mitochondria increases life span of C. elegans by decreasing the mitochondrial function. Mutation in the age-1 increased lifespan by two folds. This mutant worm had increased catalase and Cu/Zn SOD activities which may account for the increased resistance to the paraquat, a superoxide generating chemical. The mutants C. elegans support the view that the levels of oxidative stress is one of the important determinant factors in determining the length of lifespan.Superoxide anions are produced enzymatically outside the mitochondria by different oxidases and nonenzymatically inside the mitochondria. SOD detoxifies superoxide to hydrogen peroxide , which is converted to water and oxygen by catalase. There are five superoxide dismutase isoforms SOD-1, SOD-2, SOD-3, SOD-4 and SOD-5 in C. elegans. However, in most organisms there are only 3 SODs. SOD-1 is present in the cytoplasm and represents the majority of SOD activity, whereas SOD-2 and SOD-3 are present in mitochondria.

As we collect soil metagenome sequence data, we need to improve how we mine such datasets. For example, the way we conduct BLAST searches might overlook valuable information, while the unassembled reads might be too short for annotating genes with confidence. Thus, we might well need to develop new assembly and annotation algorithms. Another challenge is how to integrate different kinds of omics data, including metatranscriptomics and metaproteomics, to better understand functional processes of soil microbial communities. Metagenome sequence data, while informative, provides information about genes with the potential for being expressed, but cannot determine which ones are functional. Also, because we sequence total DNA, it is not possible to distinguish genes from actively growing cells from those in dormant or dead cells. Perhaps some analyses should be reserved for that fraction of DNA from active community members—for example, by extracting DNA that is allowed to incorporate stable isotopes or bromodeoxyuridine during replication. Ultimately,large pots plastic combining these approaches should enable us to gain a better understanding of which microbes are alive and active, and which enzymes and pathways function in soil microbial communities under different conditions.

Then we can begin to truly comprehend soil microbial communities from the microscopic to the global scale.Traumatic brain injury accounts for approximately 90% of brain injuries, and is associated with cognitive dysfunction and long-term disability. As a result of domestic incidents, military combat, traffic accidents and sports, TBI can compromise broad aspects of neuronal function. Patients often experience problems in the domains of learning, memory and affective functions that can profoundly influence quality of life. Existing therapeutic strategies for TBI have not been successful in counteracting the heterogeneous TBI pathology nor improving the quality of life of patients. Hence, identifying interventions with broad applicability seems necessary for effective management of TBI. Dietary polyphenols have significant positive effects on brain health via protecting neurons against injury and enhancing neuronal function. Evidence supports the neuromodulatory effects of flavonoid-rich blueberry, particularly in promotion of brain plasticity, and counteracting behavioural deficits. In the United States, demand for blueberries has increased, with 2017 fresh per capita consumption of 1.79 pounds/person. Several reports indicate that blueberry dietary supplementation improves memory, learning and general cognitive function, and protects against neuronal injury associated with stroke.

Moreover, it has been shown that blueberries possess potent antioxidant capacity through their ability to reduce free radical formation or upregulating endogenous antioxidant defenses. These studies suggest that blueberry supplementation can have the potential to be used to overcome the broad pathology of TBI. Given the lack of information about the effects of blueberry intake immediately after TBI, we have performed studies to assess the effects of blueberry extracts during the acute phase of TBI. Evidence suggests that TBI is characterized by dysfunction in synaptic plasticity, elevated levels of free radicals, plasma membrane dysfunction, which can contribute to the behavioural dysfunction. Oxidative stress is part of the pathology of TBI and compromises neuronal function. In particular, excessive free radical formation leads to accumulation of lipid oxidation by-products such as 4-hydroxynonenal with subsequent impairments in plasma membrane fluidity, receptor signaling across the membrane to deteriorate synaptic plasticity and reduce neuronal excitability. Deficiencies in brain derived neurotrophic factor reduce the brain plasticity necessary to cope with the effects of TBI. BDNF activates cAMP-responsive elementbinding protein , a multifaceted transcriptional regulator involved in synaptic plasticity essential for learning and memory. BDNF is known to bind to TrkB receptors, leading to activation of Ca2+/calmodulin-dependent protein kinase II , required for synaptic processes involved in behaviour. Several observations indicate that the flavonoids exert action through modulation of signaling pathways to promote synaptic and neuronal function.

Accordingly, in the current study, we investigated whether blueberry supplementation would counteract TBI pathology by involving BDNF related pathways involved in synaptic plasticity and oxidative stress to influence cognitive behaviours. Freeze-dried high bush whole blueberry fruit powder . This blend contained bio-active phytocompounds , and other macro- and micronutrients . Diet supplemented with 5% w/w BB was mixed with pulverized standard rodent chow . 1.6% fructose, 1.45% glucose and 0.0009% vitamin C were mixed with the standard rodent chow to match the levels of sugars in the BB supplemented diet and used as the rodent control diet . Sprague–Dawley male rats were purchased from Charles River Laboratories at 10 weeks of age and were acclimatized for vivarium 1 week prior to commencement of experimental procedures. Rats were housed in environmentally controlled conditions with 12-h light/dark cycle in a controlled room with free access to food and water. All procedures were approved by the University of California at Los Angeles Chancellor’s Animal Research Committee and were conducted with adherence to the guidelines set out by the United States National Institutes of Health Guide for the Care and Use of Laboratory Animals. After acclimatization, rats underwent fluid percussion injury or sham surgery and were pair housed to a specific diet group with either regular diet or blueberry supplemented diet for 2 weeks immediately. The BB dose was chosen based on previous in vivo studies which demonstrated that administration of blueberry offsets oxidative stress and reverses cognitive impairment. The groups were: Sham-RD as control group, TBI-RD, and TBI-BB. The rats were subjected to fluid percussion injury or sham surgery. Rats were subjected to learning on Barnes maze at post-injury day 14 for 5 days, and after a 2-day interval, memory was assessed at PID 21. Rats were tested for anxiety-like behaviour on elevated plus maze on PID 22 . All behavioural assessments were conducted between 9:00 and 13:00 hours. Rats were provided with diets prepared daily and fed ad libitum in powder form. To determine the voluntary food intake , food was weighed daily to measure consumption in each cage. Since the rats were pair-housed, food intake was divided by two to yield an approximate intake/rat.We employed our standard lateral fluid percussion injury protocol as described earlier. Briefly, 3% isoflurane was provided in a chamber , and then maintained with 2–2.5% isoflurane via nose cone while rats were in a stereotaxic frame. Body temperature was controlled by a heating pad. Under aseptic surgical conditions, a midline skin incision was made to expose the skull. Using a high-speed drill ,square planter pots craniotomy was made 3.0 mm posterior to bregma and 6.0 mm lateral of midline to expose the intact dura. A hollow plastic injury cap was placed over the craniotomy, secured with dental acrylic cement and was later filled with 0.9% saline. When the dental cement hardened, the anesthesia was discontinued and the rat was attached to the FPI device via the head cap. At the first response of hind-limb withdrawal to a paw pinch, rats received a moderate fluid percussion pulse . Upon resumption of spontaneous breathing the head cap was removed and the skin was sutured. Neomycin was applied on the suture and the rats were placed in a heated recovery chamber to be fully ambulatory before being returned to their cages. The sham animals were prepared using the identical surgically procedure but without the fluid pulse. Barnes maze testing was performed 2 weeks after experimental TBI with two trials per day with a 5-min test period . For learning assessment, rats were given two trials per day for five consecutive days at approximately the same time every day. Subsequently, memory retention was assessed at post-injury day 21. Latency to finding the escape hole and search strategies were analyzed for each trial.

Three search strategies were identified using following categorization: spatial, peripheral, and random using data recorded with AnyMaze software. EPM test was performed to assess anxiety-like behavior 2 weeks after experimental TBI with two trials as described in the Supplemental Information. Specifically, rats were individually placed in the closed arm of the EPM apparatus and permitted free exploration for 5 min during which their movements were camera recorded. The behaviors scored were time spent and number of entries into the open arm using automated video tracking system. EPM testing was conducted after Barnes maze memory test. Upon completion of the experiment, hippocampal tissues were harvested frozen in dry ice, and stored at –80 °C until use for immunoblotting. The left side hippocampus were homogenized in a lysis buffer containing 20 mM Tris–HCl , 137 mM NaCl, 1% NP40, 10% glycerol, 1 mM phenylmethylsulfonylfluoride , 10 μg/ml aprotinin, 0.1 mM benzethonium chloride, 0.5 mM sodium vanadate. Total protein was then determined using a BCA Protein Assay kit , using bovine serum albumin as standard. Equal amounts of protein were separated by sodiumdocecylsulphate-polyacrylamide gels and then transferred onto polyvinylidene difluoride membranes . Membranes were probed with anti-actin or anti-BDNF, anti-pCREB, anti-CREB, anti-CaMKII, anti-4- hydroxynonenal followed by secondary antibody . Immunoreactive proteins were visualized using enhanced chemiluminescence reagents . Band intensities were quantified using Image J32 Software. β-actin was used as an internal control for normalization western blot such that data were standardized according to β-actin values. Blots for each experimental group were normalized to ShamRD values within the same gel. Protein data are expressed as mean ± standard error of the mean . Body weight data expressed as mean ± standard deviation. Statistical analysis was performed by software GraphPad Prism 7.04. A level of 5% probability was considered as statistically significant. The Barnes maze learning data analysis were analyzed by repeated measures analysis of variance . Protein results are expressed as percentage of Sham-RD group.In the present study, we found that BB supplementation can attenuate important aspects of the acute TBI pathology. We report that BB supplementation immediately following TBI mitigates behavioural deficits in spatial learning and memory. BB supplementation counteracted the effects of TBI on proteins associated with the action of BDNF on plasticity and behaviour. In addition, BB supplementation counteracted the increase of the end product of lipid peroxidation, 4-HNE. The results showing that markers of neuronal plasticity and lipid peroxidation change in proportion to memory performance suggest a possible association between these molecular parameters and behaviour. Taken together, the present findings emphasize the beneficial effects of BB supplementation in fostering brain plasticity in the TBI pathology. In agreement with previous reports, we found that TBI impairs spatial learning as evidenced by an increase in latency in the Barnes maze, while BB supplementation decreased latency time to find the escape hole at each training day. We assessed the use of spatial learning strategies in our paradigm to provide a complementary measure of cognitive function less dependent on motor behaviour. Interestingly, we found that BB supplementation appeared to counteract a lost capacity of TBI rats to employ spatial leaning cues. This information together with results of the shorter latencies strongly suggest that BB supplementation protects TBI animals from a loss in spatial learning performance. In this regard, recent functional neuroimaging study in humans has established a connection between BB intake and cognitive function. Further, in the elevated plus maze test, rats exposed to TBI showed a tendency to reduce time spent in the open arms, which encompasses with clinical reports that psychiatric disorders are often observed in TBI patients. TBI-induced behavioural deficits probably stems from the impairments in BDNF-TrkB signaling that has been implicated in various cognitive and affective disorders. We cannot ascertain the cellular identity of the reported protein alterations. Although neuronal cells are the primary locus for learning and memory processing, non-neuronal cell types such as astrocytes and microglia can also contribute to these alterations. Moreover, it known that astrocytes and microglia provide support to synaptic transmission that is fundamental for neuronal function involved in cognitive processing.In the present investigation, we also found that TBI significantly reduced levels of hippocampal BDNF, and that BB dietary supplementation normalized these the levels. Previous report indicated that deficiencies in BDNF signaling is associated with impairments in cognition. Alternatively, cognition is strongly reliant on long-term potentiation and hippocampal BDNF, and the interaction between BDNF and its tyrosine kinase receptor is required for induction of LTP. Previously we have shown the protective effects of BDNF on the TBI pathology. Presently, our findings show that BB supplementation counteracted the BDNF reduction induced by TBI, paralleling improvements in cognitive function. It is well established that BDNF regulates synaptic plasticity and learning through interaction with the transcription factor CREB. Interestingly, our results also showed that BB supplementation normalized levels of CREB in TBI animals, and that these changes were proportional to changes in BDNF levels.

Long-term climatic cycles, such as El Niño Southern Oscillation events, could also play a role, particularly in rainfed systems that we may expect to exhibit stronger variability than their irrigated counterparts. ENSO phases have been shown to induce synchrony in masting systems, but knowledge of ENSO effects on crop plants is largely limited to annual crops. As a primary source of climate variation in Brazil and Iran, ENSO could be a cause of periodic yield in Brazilian tangerine and Iranian apricot . Climate and pest cycles may also interact to produce complex longer-term cycles in crops. If future work confirms a biological basis of long cycles in some crops at a national scale, it would be profitable to understand the extent to which these longer-term crop fluctuations reflect exogenous forcing versus endogenous feed backs . In conclusion, we have found that perennial crops frequently exhibit alternate bearing even at a national scale and that this is especially pronounced in wind-pollinated crops. This pattern is remarkable, given the general assumption that management practices have come to outweigh the ecological drivers that would synchronize country-wide production. Our results suggest that historic yield data present a thus far underused resource for further analyses on the mechanisms of reproductive synchrony across time, space and taxa.

Future work could explore the intraspecific and intraregional differences in synchrony and the degree of overlap and divergence between patterns in natural and managed systems. We encourage strengthened collaborations between theoretical ecologists,drainage gutter applied ecologists and horticulturalists for the mutual benefits of achieving an enhanced understanding of the mechanisms of synchronous interannual variability and promoting stable yields and farmer livelihoods.Bees and other flower-visiting animals provide essential pollination services to many US crops and to wild plant species . Bees contributed an estimated 11% of the nation’s agricultural gross domestic product in 2009 , equal to $14.6 billion per year . Of this, at least 20% is provided by wild pollinators that depend on suitable land for nesting and foraging . As the consumption of specialty fruit and vegetable crops has grown , the demand for pollination services has increased. However, the supply of managed honey bees has not kept pace , due to management challenges and colony losses over the last decade . There is growing evidence that wild, unmanaged bees can provide effective pollination services where sufficient habitat exists to support their populations . They can also contribute to the long-term stability of crop pollination, thereby reducing the risk of pollination deficits from variable supply or activity of honey bees . As a result, wild pollinators should be integrated into crop pollination management plans as a supplement or alternative to managed bees . Despite the agricultural importance of wild bees, there is increasing evidence that multiple species are declining in range or abundance. Some of the most important crop pollinators, such as bumble bees , have declined over past decades in the United States . Among the numerous threats to wild bees, including pesticide use, climate change, and disease , habitat loss seems to contribute to most observed declines .

Indeed, a National Research Council committee on the status of pollinators in North America reported that conserving and improving habitats for wild bees is important for ensuring continued pollination services and food security . Recognizing both the growing need for pollination services and increasing threats to wild bees, a recent presidential memorandum called for a national assessment of the status of wild pollinators and available habitat in the United States . The resulting report sets a goal of 7 million acres of land for pollinators over the next 5 y . However, there has been no assessment at the national level of the current status of native pollinator habitat, where and at what rate this habitat is being degraded, and the impact of these changes on bee populations and the pollination services they provide. A national assessment is challenging because plant–pollinator interactions and dynamics occur at relatively fine spatial scales. Wild bee populations are largely determined by the spatial distribution of habitat resources within their foraging range , and this varies from ∼100–2,000 m . Accordingly, most of our understanding of native bee populations is at the scale of landscapes and local sites. Several field-based assessments of habitat resources for native bee species have been developed at landscape scales . However, the required cost and time to scale this type of field assessment to cover all habitat types and bee species nationwide is logistically challenging and prohibitively expensive. When field observations are lacking, careful use of expert derived data has been shown to provide informative estimates that enable habitat assessments , including studies on pollination . Use of expert opinion may therefore be an efficient path to an initial nationwide assessment of pollinator habitat and abundance in the absence of consistent data across different land categories.

Such an approach must include careful treatment of uncertainty that may arise from differences in expertise among regions, authorities, taxa, and so on . Indeed, a robust analysis of uncertainty, and its implications for assessment findings, is a useful result in itself. It can help orient research toward addressing the most important gaps in our national knowledge of wild bees and their importance for crop pollination. Here, we use a published model of bee abundance and expert knowledge to assess the status, trends, and impact of wild bee abundance and associated uncertainties across the coterminous United States. The spatially explicit model predicts a relative index of wild bee abundance based on local nesting resources and the quality of surrounding forage . We parameterize the model with expert-derived estimates of nesting and forage quality for each of the main land-use types in each of the major ecoregions to construct a probability distribution for each parameter that captures estimates by multiple experts and their uncertainty. We first validate model predictions with bee collections and observations from a variety of landscape settings. We then map bee abundance, its uncertainty, and the agricultural demand for pollination across the United States to address the following questions: What are the current status and trends of wild bee abundance across the coterminous United States? What land use changes have driven these trends over a 5-y period ? Which regions and crops experience relatively low bee abundance compared with crop pollination demands? How does uncertainty in our knowledge affect these predictions? Responses to these questions will inform future research efforts and policy decisions to conserve native bees at the national level and can help guide a coordinated and ongoing nationwide assessment of wild bees.Our model predicts generally high abundances of wild bees in areas rich in resources such as chaparral and desert shrublands,plastic gutter intermediate abundances in temperate forest and grassland/rangelands, and lower abundances in most agricultural areas . Patterns of wild bee abundance and expert uncertainty seem correlated . In fact, whereas most areas with low bee abundance also present low uncertainty, only 5% of areas with high bee abundance have low uncertainty. This suggests that experts are more individually or collectively certain about uniformly poor bee habitats than they are about higher-quality habitats , which can vary in quality over time and space . Between 2008 and 2013, wild bee abundance was consistent in 67% of the US land area . However, our model indicates decreases in 23% of the United States , and these decreases were highly likely in 9% of the United States . Most of the areas of likely decrease occurred in agricultural regions of Midwestern and Great Plains states and in the Mississippi river valley. Eleven states [Minnesota, Texas , Wisconsin , South Dakota , North Dakota , Illinois, Missouri, Nebraska, Oklahoma, Kansas, and Louisiana] collectively accounted for 60% of the areas of predicted decrease in wild bee abundance. Over the 5-y period in these states, corn and grain cropland increased 200% and 100%, respectively, and mostly replaced grasslands and pasture . Bee abundance increased in 10% of the United States and the increase was highly likely in 3% of the country .

Areas of likely increase in bee abundance were found in northern ND, eastern Washington and Pennsylvania , southern Montana, parts of several states in the Great Plains, and in southeastern coastal areas . In these areas, grasslands, pastures, and corn/soy fields were converted to higher-quality habitat, such as shrublands or fallow crop fields .Bee abundance maps can be interpreted as the potential “supply” of pollination services from wild bees. To compare this measure of supply to potential agricultural demand, we calculated the area of pollinator-dependent crops, weighted by each crop’s degree of pollinator dependence, for each US county in 2013 . By comparing the two maps, we identified counties with relatively high supply of wild bees and relatively low demand and, conversely, where high demand occurs in counties with relatively low supply . We identified 139 counties where high demand and low supply coincide and 39 counties where this difference was particularly extreme . All of the 139 counties with a pollinator disparity had relatively low uncertainty for 2013 bee abundance , which indicates that there is high confidence in this mismatch. These counties tend to contain either a significant percentage of area that consists of highly pollinator-dependent crops [e.g., almonds, blueberries, and apples in California , Oregon, and WA, respectively] or large amount of less-dependent crops . To examine changes in the relationship between wild bee supply and pollination demand, we combined the two trend maps . We found that 106 counties have simultaneously experienced increases in demand for pollination services and decreases in wild bee abundance . This represents 54% of the 195 counties that have experienced substantial changes in pollination demand . In 27 of these counties, declines in supply were highly likely , whereas in the remaining 79 counties declines were less certain . In counties of West Coast states and Michigan, increases in demand were mostly driven by increases in specialty crops such as almonds, cherries, blueberries, apples, watermelons, and squash. In contrast, demand increases in the Great Plains and Mississippi Valley were driven by increases in crops, such as sunflower, canola, soybeans, and cotton, with moderate to low pollinator dependency. Trends in our measures of supply and demand vary widely among individual crops . Most crops that require animal pollination have expanded in area between 2008 and 2013, whereas the predicted supply of wild bees in many of these cropped areas has declined. Specialty crops, such as pumpkins, blueberries, peaches, apples, and watermelons, are among the crops that present the strongest mismatch between changes in supply and demand. Others, such as canola, have experienced increases in both supply and demand. Of particular concern for future abilities to meet pollination demands, crops that are most dependent on pollinators tend to have experienced simultaneous declines in supply and increases in demand.Our study is the first to our knowledge to map the status and trends of wild bees and their potential impacts on pollination services across the coterminous United States. By combining a spatial model with expert knowledge, we find highly heterogeneous patterns of both predicted abundance of wild bees and our uncertainty regarding those predictions. We also identify counties and crops of potential concern, where declines in wild bee abundance oppose increased need for crop pollination. These analyses form an important step toward a nationwide understanding of the status of wild pollinators. They can also help focus attention and future research toward regions of high uncertainty and to direct management efforts to areas of major concern. Our mapped index of bee abundance clearly shows that areas of intense agriculture are among the lowest in predicted wild bee abundance. Our predictions are also relatively certain in these areas . This reflects consensus among experts about the low suitability of intensively managed agricultural land for wild bees and is supported by an abundance of previous research on the negative effects of intensive agriculture on bee populations . Areas of bee abundance where declines are most certain tend to have experienced additional conversion of natural land covers to crops, especially corn . These results reinforce recent evidence that increased demand for corn in bio-fuel production has intensified threats to natural habitats in corn-growing regions . For example, a recent land-use simulation found that expansion of annual bio-fuel crops could reduce pollinator abundance and diversity at the state level .

The walnut industry submitted its research results for a heart health claim to the U.S. Food and Drug Administration , and the almond industry submitted its as part of a petition filed by the International Tree Nut Council Nutrition Research and Education Foundation to the FDA for a heart health claim for nuts. The FDA approved a qualified health claim for walnuts, and another for almonds and other selected nuts, on July 15, 2003, which states: “Scientific evidence suggests but does not prove that eating 1.5 ounces per day of as part of a diet low in saturated fat and cholesterol may reduce the risk of heart disease.” Each of these four commodity groups has used news releases and public relations to publicize the nutrition and health benefits of their products. The messages for walnuts and avocados have been communicated almost entirely through third parties such as magazines, newspapers, doctors, nutritionists or other credible sources, rather than paid advertising. The advertising emphasis for walnuts and avocados has been on quality, taste and recipes. The Almond Board of California initially relied on public relations to disseminate its message on the health and nutritional benefits of almonds. Following FDA approval of their qualified health claim, their research results were incorporated into paid advertising and promotion, almost always featuring a health message.

Likewise, in 2003 the California Strawberry Commission introduced a promotion campaign called “Be Well — Get the Red Edge” , which targets health and nutrition professionals,greenhouse ABS snap clamp and consumer and trade media. Commodity groups have found that they can stretch their promotion budgets by partnering with other groups. After FDA approval of the qualified heart health claim for nuts, the Almond Board of California partnered with the American Heart Association and now makes liberal use of the AHA logo in almond advertising. The California Walnut Commission formed a partnership with the Spanish Heart Foundation, and distributed more than 40,000 brochures and samples during “Heart Week” in Spain. The foundation also includes recipes and other information concerning California walnuts on its Web site. The California Avocado Commission is also leveraging its research results by partnering with health organizations, including the American Diabetes Association, UCLA Nutrition Education, American Association of Diabetes Educators, American Dietetic Association and IDEA Health and Fitness Association. Imports can easily create a “free rider” problem for U.S. commodity programs. California avocado producers spent more than $182 million on market development between 1961 and 2003 . They were understandably upset to see producers in other countries taking advantage of state promotional efforts when the national market share of imported avocadoes increased from less than 3% prior to 1990 to about 34% from 1998 through 2002. With the 2002 passage of the Hass Avocado Promotion and Research Order , all Hass avocados sold in the United States, including imports, are assessed 2.5 cents per pound to fund advertising, promotion, research and data dissemination.

Increased avocado demand due to HAPO promotion will offset much of the price impact of increased imports, and importers should enjoy attractive returns from their promotion dollars. Carman estimated that returns for importers’ spending on advertising and promotion ranged from $2.09 to $6.31 per dollar spent, depending on the level of imports and the effectiveness of Hass avocado advertising.An important and often overlooked benefit of mandated marketing programs, in addition to having an organized commodity group, is the value of the information they gather, organize and disseminate. A first-of-its-kind innovation for commodity groups was the establishment, by the Hass Avocado Board , of a Web-based program designed to exchange crop and marketing information among 100 packers and over 20,000 producers from the five HAB members — California, Chile, Dominican Republic, Mexico and New Zealand. The HAB Web site notes: “The program goal is to develop collaborative strategies to achieve an orderly flow of the 750 million pounds of fruit sent annually into the U.S. marketplace.” This Web-based program collects, tracks, analyzes and disseminates information relevant to selling Hass avocados in the U.S. market. It provides all players in the U.S. market with 24-hour access to critical market data that drives decisions about growing, shipping, distribution and marketing. The HAB market information program is an exciting development in produce marketing, made possible by recent worldwide developments in communication networks. It provides a dramatic reversal of trends that have reduced the availability and timeliness of market and price information for produce markets.

The widespread availability of marketing information and data is a theoretical requirement for competitive markets. It will be interesting to see to what degree improvements in marketing efficiency made possible through HAB flow to producers funding the program. If successful, the program template can be extended to other commodities and countries, with benefits flowing to producers and consumers worldwide.Drosophila suzukii , commonly known as the spotted-wing drosophila, is an invasive pest of small fruit and berry production native to Southeast Asia. Starting in 2008, this species underwent a rapid geographic range expansion, spreading across North America, South America, and Europe. Following its arrival to the USA, growers in California, Oregon, and Washington reported yield losses of up to 50% in raspberries and blackberries, 40% in blueberries, and 33% in cherries. Unlike other Drosophila species that typically breed on overripe and rotting fruit, D. suzukii females readily lay eggs into firm, still-ripening fruit using specialized serrated ovipositors. After hatching, larvae continue to feed and develop within the fruit, causing rapid decay and increased susceptibility to subsequent infestation by other drosophilids. Efforts to control D. suzukii and prevent infestation have led to a drastic increase in the use of broad-spectrum insecticide applications in susceptible host crops. Specifically, three main classes of insecticide–organophosphates, pyrethroids, and spinosyns–have shown high efficacy and now form the basis of most conventional D. suzukii management programs. Carbamates and some diamides have also shown moderate efficacy against D. suzukii, but the use of these materials is more limited throughout North America and Europe. The continued reliance on chemical sprays to control this pest, combined with its high fecundity and rapid development time, have elicited concerns that insecticide resistance could quickly develop. For organic growers, the risk of resistance is even greater. Currently, spinosad is the only insecticide approved by the Organic Materials Review Institute with high efficacy against D. suzukii, meaning that growers must rotate spinosad sprays with low-efficacy materials such as pyrethrins and azadirachtin in order to comply with label requirements. Consequently, D. suzukii in organic fields are primarily challenged by only one class of insecticide further increasing the risk of resistance. Indeed, recent work by Gress and Zalom reported low to moderate levels of spinosad resistance in D. suzukii collected from commercial caneberry fields near Watsonville, CA. Specifically, this study utilized dose-response analysis of adult D. suzukii to quantify the degree of resistance and found that Watsonville females exhibited LC50s 5–8 times higher than females from a second, untreated location in California. This finding demonstrates an immediate need to expand our understanding of resistance evolution in D. suzukii and to develop novel tools that aid in the early detection and assessment of resistant populations so that appropriate management actions can be implemented. When decreased susceptibility to an insecticide is first detected in a pest field population,snap clamps ABS pvc pipe clip laboratory selection studies, often referred to as resistance risk assessments, represent a critical first step towards understanding the potential for resistance to develop. These studies are performed by exposing large numbers of individuals from the suspect field population to moderately lethal concentrations of insecticide for multiple generations such that the most susceptible individuals are eliminated and the most tolerant survive. Dose-response analyses are performed both before and after implementing the selection protocol, and the change in LC50 is used to quantify the response to selection and better understand the potential for resistance to develop. This approach, however, is not without its limitations. For example, because laboratory colonies contain limited genetic variation relative to large field populations, these studies can understate the evolutionary potential for resistance to develop in a field setting.

Conversely, selection imposed in laboratory settings may be more intense and consistent than in the field, leading to possible overestimation of resistance development. Nevertheless, an increase in resistance due to laboratory selection provides unambiguous evidence that further loss of susceptibility in the field is possible. Additionally, if laboratory selection is successful and resistant colonies are generated, these strains can assist researchers in the development of novel tools for detecting, monitoring, and managing resistance in the field. For example, resistant colonies can be used to identify the molecular mechanism that produces the resistant phenotype, and these markers can, in some instances, be used to track spatiotemporal patterns of resistance development in the field. Laboratory-generated resistant colonies can also be used to assess whether insecticide resistance in a particular population is associated with fitness costs , and this information can be used to develop programs to manage resistance in the field. The presence of such costs under benign conditions would suggest that efforts to temporarily eliminate the target insecticide from spray programs could successfully maintain susceptibility in the field, at least in the short-term. After decreased spinosad susceptibility in the Watsonville, CA population was identified, Gress and Zalom performed a resistance risk assessment by continuing to expose adults from this population to a discriminating dose of spinosad using a standard glass vial bio-assay protocol developed for D. suzukii . After implementing this technique for 5 generations, Gress and Zalom found that LC50 values increased by 86% for males and 49% for females. This selection protocol, however, is highly labor intensive and costly, making it impractical for long-term selection programs which are needed to generate and maintain resistant lines. Additionally, this method requires that researchers kill off much of their adult population each generation, increasing the risk that selection lines will be lost in the process. Here, we describe a simple larval bio-assay that overcomes these challenges and requires little effort or cost beyond what is already needed for basic laboratory stock maintenance. We then perform a series of larval dose-response bio-assays to calibrate the selection protocol and identify baseline susceptibility for three commonly used insecticides. Finally, we implement the larval bio-assay protocol to perform resistance risk assessments using the same Watsonville, CA field strain previously tested by Gress and Zalom. Changes in the susceptibility at both the larval and adult life stages were quantified, and results from this study are compared to those obtained with the original adult selection method.First, groups of n = 20 mated females were transferred into plastic 6 oz Drosophila stock bottles containing fresh Bloomington standard Drosophila cornmeal diet. Females were allowed to lay eggs for four days before being transferred into new bottles with fresh diet. Once females were removed, 400 μL of insecticide solution was pipetted onto the surface of the diet, and each bottle was shaken sideto-side to distribute the solution. This ensured that all larvae were uniformly exposed to the insecticide, both through external contact and internally while feeding on the treated diet. Control bottles were treated with either ddH2O or acetone, depending on the insecticide treatment , but otherwise were handled using the same protocol as experimental bottles. All bottles were then re-plugged with a cotton stopper and stored in a climate-controlled walk in chamber at 22–23˚C and 14–10 light-dark cycle. Because the first larvae typically crawl out of the diet to pupate after 6 days under these conditions , treatment timing is critical to ensure that all larvae are exposed. Bottles were monitored for up to 18 days following treatment for the emergence of adult D. suzukii, and all newly enclosed adults were transferred to fresh bottles and counted.To quantify baseline susceptibility of D. suzukii larvae, we used the larval bio-assay protocol to perform dose-response analyses for three insecticide AIs, each representing a different insecticide class, commonly used against this pest. This work was conducted using a susceptible laboratory strain of D. suzukii originally collected from USDA Wolfskill Germplasm Repository in Winters, CA in fall of 2017. Wolfskill is an experimental, mixed-fruit orchard, and crops at this location receive no insecticide treatment. This strain of D. suzukii was previously used as a susceptible control by Gress and Zalom when quantifying resistance in the Watsonville population. The three formulated insecticides used in this study were malathion , spinosad , and zeta-cypermethrin .