Horticulture

As well as direct interaction and disturbance, avoidance of interspecific chemical cues and resource competition have the potential to alter pollinator foraging movements. Global human population growth is putting greater pressure on agricultural production. There is concern over how to meet the increasing demand for food, while at the same time safeguarding ecosystems and biodiversity. In the future, land currently under agricultural production will have to be more intensively managed to increase yields and/or more land will have to be converted to agriculture. Given the negative impact agriculture has already had on biodiversity, it is important that future steps to increase production be made environmentally sustainable. In the last 50 years, the fraction of agricultural production requiring biotic pollination has more than tripled. When compared with crops that are not pollinator-dependent, those that are moderately pollinator-dependent have shown slower growth in yield and faster expansion in area from 1961 to 2006. Almond is a mass-flowering, varietally self-incompatible crop species, hydroponic nft gully highly dependent on biotic pollination. Almond orchards are generally planted with alternating rows of two or more varieties.

Planting a single variety per row facilitates harvest, but complicates pollination because pollen must be transferred between rows to achieve fruit set. To allow for management activities, trees between rows are further apart than those within the same row . Apis mellifera tend to forage within a tree and then move down the same row, probably because less effort is required to move to the next tree in the same row or because the rows act as visual markers that influence movement. This foraging pattern means A. mellifera tend to move more incompatible pollen, limiting their pollination effectiveness. In almond, we investigated whether the presence of nonApis bees affected the behaviour and pollination service of the dominant pollinator species, A. mellifera. Often almond orchards are isolated from natural habitat and non-Apis bees can be completely absent. Therefore, we were able to compare A. mellifera behaviour and pollination effectiveness in diverse bee communities with orchards lacking non-Apis bees. Here, we refer to pollinator effectiveness as the probability an ovule is fertilized following a single visit. We complemented our intensive field sampling with observations in a controlled cage environment, where A. mellifera were introduced along with the blue orchard bee Osmia lignaria Say. We hypothesized that where non-Apis bees were present, such as in sunflower, interspecific interactions would cause A. mellifera to more frequently move between rows. We further hypothesized that an increase in between-row movements by A. mellifera would increase their pollination effectiveness and increase fruit set .

Our findings show that increased pollinator diversity can synergistically increase pollination service through species interactions that alter the behaviour and resulting functional quality of a dominant pollinator species. This highlights a largely unexplored facilitative component of the benefit of biodiversity to ecosystem services. Total bee visitation rates were similar between orchard types, and the visitation rate of A. mellifera was lower in orchards with non-Apis bees. However, A. mellifera pollination effectiveness was greater and fruit set was higher when non-Apis bees were present. Apis mellifera increased their proportion of movement between tree rows when non-Apis bees were present, thereby improving pollination effectiveness. More successful pollen tube growth translated into significantly higher fruit set in the orchards where non-Apis bees were present. The greater proportion of between row movements by A. mellifera individuals most probably resulted in the deposition of more compatible pollen, since pollen from the same variety generally does not set fruit . This synergistic effect of the presence of non-Apis bees suggests that maintaining biodiversity in agricultural ecosystems could provide unrecognized benefits, and it offers exciting opportunities for the integration of more diverse pollination systems to improve the longer-term sustainability of crop production for almond and similar crops. Unlike in sunflower, very few direct interactions were observed between A. mellifera and non-Apis bees. One potential mechanism for the increased proportion of inter-row movement of A. mellifera in the presence of other non-Apis bees is linked to resource depletion. Because some non-Apis bees can fly at lower temperatures than A. mellifera, and therefore earlier in the day, it may be that if A. mellifera begin foraging and find flowers have already been depleted then they increase the distance of their foraging flights. Alternatively, it may be related to the scent marks left by non-Apis bees on the flowers.

Apis mellifera have been shown to avoid visiting flowers that have been marked by bumble bees, and it may be that as well as avoiding the flowers they also shift their foraging location. Given that A. mellifera tend to move down the same row, if a response to perceived resource competition is to shift foraging locations, this could be expected to involve movement across rows. However, at present the mechanism for the increased between-row movements when non-Apis bees are present is unknown. In the controlled cage environment, the steeper increase in the number of pollen tubes initiated with increasing pollen deposition in the mixed cage and the higher fruit set supports the findings from the open orchards that more compatible pollen is being moved when pollinator communities are diverse. Because there was only one cage per treatment, the data from the cages are only descriptive, but they do support the findings from the open orchards. Future work should attempt to replicate similar treatments. The greater fruit set in the mixed cage when the visitation rate was lower than in the A. mellifera cage may be due to the slightly higher proportion of between-row movements in the mixed cage. Although the differences in movement were not great, the pollen tube data suggest that more pollen was moving between varieties in the mixed cage. The size of the cages limited the opportunity for between-tree flights. This and the relatively low visitation rate of O. lignaria in the cages compared with the open orchards may explain why the difference in pollen tubeformation and fruit set between the cages was less than in the open orchards. The price of California almond in 2011 was approximately $1.79 per pound and production was estimated at 2670 lbs per acre . If a 5 per cent increase in fruit set as calculated from the controlled cage environment translated into an equivalent increase in production, farmers would make $239 more per acre . This estimate from cages may be a lower bound compared with open orchards. Sampling effects and complementarity have been the primary explanations for a positive relationship between biodiversity and ecosystem function. Here, we show a different mechanism, possibly due to interspecific competition, where community composition alters the behaviour of a service-providing organism with a positive knock-on effect for the ecosystem service. Our results show alterations in A. mellifera foraging behaviour when a diverse community of other bees are present, and suggest almond yield can be increased by encouraging wild bees in the orchards. As such, dutch buckets for sale natural habitat near almond orchards should be conserved to protect wild bee communities. The availability of A. mellifera is not predicted to increase at the same rate as demand for their services in agriculture. Thus, increasing the pollination effectiveness of A. mellifera and conserving wild pollinator communities could help increase crop yields. The synergistic combination of A. mellifera and non-Apis bees represents a sustainable way to improve crop pollination services, but the generality of such effects still need to be tested across multiple crop systems.The two coffee species of commercial value, Coffea arabica and Coffea canephora , both originate from Africa; the former has generally preferred taste qualities, and the latter exhibits higher yield and pest resistance . Coffea arabica dominates global coffee landscapes, accounting for 60% of the total coffee volume produced . Although coffee’s center of origin lies in Ethiopia, major global dispersal of the bean occurred when Arab and European traders introduced the beverage to Western Europe in the early 1500s . By the latter half of the 1800s, coffee plantations of both C. arabica and C. canephora flourished throughout the American tropics; by the 1970s, coffee’s cultivation dominated more than 8.8 million hectares of tropical landscapes. Between 1970 and 1990, the global coffee area and average yields increased by 25% , and global production increased by 58% .

Interestingly, although the global area decreased to 10.2 million ha between 1990 and 2010 , production still climbed 36%, which is evidence of an overall intensification in several key countries , coffee abandonment in others , and the rapid expansion of high-yield coffee in new countries . Brazil, for instance, saw a 112% jump in production with only a 12% increase in coffee area between 1996 and 2010, growth spurred by intensification that resulted in an 89% yield increase over that period , and recognition from coffee experts that production there has been highly industrialized . Since the mid-1980s, exports of robusta coffee have increased by 92%, led by a number of Asian countries, with Vietnam being the prime example, exhibiting hand-in-hand increases in both area and intensification . Robusta yields there soared from a historical average of 450 kg per ha prior to the 1950s to 1558 kg per ha by 2004 , more than double the global yield average at the time, which reveals that a species shift alone does not explain yield increases. Given that coffee area decreased globally by 9% between 1990 and 2010, whereas world production increased by 36%, we posit that intensification is one of the major drivers of shifting coffee cultivation practices. A closer look reveals that the shift in production between 1990 and 2010 was regional: 45% of the nations exhibiting decreases were in Africa, whereas Asian countries accounted for 35% of those with increased production . When the first comprehensive studies of coffee and biodiversity emerged in 1996, the top three producing countries were Brazil, Colombia, and Indonesia. Currently, Brazil, Vietnam, and Indonesia top the list, accounting for 57% of the 8.2 million metric tons produced in 2010. In Vietnam alone, the cultivated area increased by 731%, yields by 45%, and the total production by 1102% between 1990 and 2010 . In contrast, the past 20 years reveal coffee area declines exceeding 20% in Ecuador, Colombia, Côte d’Ivoire, Mozambique, Madagascar, Tanzania, and Rwanda . The contrasting and heterogeneous changes in global coffee cultivation result from multiple factors, including region-specific economic development patterns, political conflict, cultural practices, land values, wages, and labor. For example, deforestation accompanied increases in coffee area in Vietnam, Indonesia, Nepal, and Panama . In contrast, in places where the coffee area has declined, such as Costa Rica and Ecuador, the expansion of high-yield agriculture has caused a decrease in coffee prices, which has, in turn, resulted in the abandonment of marginal agricultural lands in combination with increased land prices due to urbanization. Higher land values due to exurbanization often displace coffee cultivation in places like Panama’s Boquete and Chiriquí regions, Costa Rica, and Guatemala— areas now popular as retirement destinations . In a number of countries, waves of political and social instability have reduced investment in coffee cultivation , but sustained global prices after 2005 have spurred expansion in other countries . In other regions, the draw of better urban wages or displacement by other cash crops, such as cacao , has reduced the area of coffee production. Despite variation in global coffee production, the majority of coffee is still produced by smallholders managing less than 10 ha of coffee , as has been documented in Asia and in Africa . Likewise, in Central America, smallholders represent 85% of coffee producers but control only 18% of coffee production lands . In some coffee-producing countries, such as Rwanda, coffee farm sizes are so small that the majority of farms are measured by the number of coffee bushes instead of in ha , whereas in many Mesoamerican smallholder farms, stand densities are as high as 6700 coffee bushes per ha . These patterns in farm size tend to shift, depending on coffee prices and government incentives, as is evidenced in Latin America, where a decrease in the number of large estates and an increase in the number of smallholders and microproducers occurred directly after the 1999 coffee crisis, when coffee prices dropped to century lows . In the Costa Rican coffee district of Agua Buena, the proportion of farmland dedicated to coffee production diminished from 52% to 24% between the years 2000 and 2009, whereas the proportion of pasture land increased from 31% to 50%, largely because of basement level international coffee prices .

The Department of Education projects that enrollments will grow by 0.7 percent in the current fiscal year. The education budget request remarks that “additional funding proposed by the Governor will allow local districts and charter schools to decide how to best improve the education opportunities of their students.” As required by Amendment 23, the inflationary increase to the State Education Fund amounts to $8.9 million. The Department of Education proposed to allocate much of this increase to “categorical programs,” which are educational initiatives tailored to specific groups of students including those with disabilities, ESL students, at-risk students, and gifted and talented students, as well as specialized programs such as technical training, health education, and school transportation. The final budget included funds to support many of these initiatives with a $78.12 million increase in spending from the prior year. This number is below the increase requested in the governor’s budget, hydroponic dutch buckets but does represent an investment in education when many competing priorities exist. In dollar amounts, the proposed increase in higher education funding is similar to that for K-12 education.

Increasing higher education spending by $86.9 million is almost a 10 percent increase from the current fiscal year’s funding level. Among the notable initiatives in the higher education budget are $73.1 million to improve college affordability and outcomes. This includes a $59.1 million allocation to state college governing boards “to address affordability for all students by moderating higher education tuition increases” . Additional funds indirectly distributed to students include $13.9 million for need-based financial aid and work-study programs and $4.0 million for the Colorado Opportunity Scholarship Initiative. The budget also includes lines for new programs such as the $1.5 million proposed for an Emergency Completion and Retention Grant, which is a one-time benefit for students experiencing an unexpected financial hardship, and $5.0 million for a new Occupational Credential Capacity Grant program, which supports “high-demand post-secondary certificate credentials.” The General Assembly elected to fund these college affordability efforts, among other programs, by providing more funding than the governor’s budget requested. The final appropriation for Higher Education was in excess of $1.0 billion, a 12.1 percent increase, which is greater than the governor’s requested increase of 9.7 percent.

In dollar amounts, this increased outlay is among the largest in the approved budget after the $115 million increase approved for Human Services. After K-12 education, health care spending represents the second largest budgetary outlay. Similar to the education budget, legislators approved health care funding at a level slightly less than Governor Hickenlooper’s request. The Department of Health Care Policy and Financing is slated to receive increased General Fund appropriations totaling nearly $100 million, despite an expected decline in the state’s Medicare caseload. Among the new spending initiatives regarding health care include $89.4 million for Medicaid, the Child Health Plan Plus , and the Medicare Modernization Act. Provider rate adjustments proposed in the budget total $10.3 million, which constitute a standard increase of 0.77 percent. The final budget increased health care spending by $81.8 million . In percentage terms, the department with the largest proposed budget increase is Personnel at nearly 17 percent. Funds were also included to hire six full-time employees to assist with the state archives and the address confidentiality program, which assists individuals who have experienced domestic violence. The legislature approved an increase about 12.5 percent, which is the fifth largest increase in the final budget. Departments with an increase in spending in excess of this amount, which were greater than the governor’s proposals, include Treasury, Public Safety, Local Affairs, and Human Services. As seen in Table 3, the largest deviation from the governor’s proposal is the substantial increase in Treasury spending. This increase is primarily attributable to $225 million in approved appropriations from a separate bill as an unfunded liabilities distribution to PERA. Conversely, the department with the largest proposed cut to its budget is Department of Labor and Employment, which the governor’s budget request proposes to reduce by 13.5 percent. The approved budget for Labor and Employment imposes a cut of 8.9 percent, which is the only department’s budget reduced by the General Assembly for this fiscal year.

While not reducing the budget from the prior year, data reported in Table 3 show that legislators ultimately provided less funding than the governor proposed for six departments, while the remaining 13 departments received funding in excess of Governor Hickenlooper’s request. Similar to previous years, other departments with large budgets include Human Services , Corrections , and the courts . The Human Services budget proposed salary increases for staff as well as imposing a 1 percent increase in community provider rates . The budget also proposed nearly $10.0 million in General Fund spending for the Office of Behavioral Health, which provides numerous services including court-ordered health evaluations and outpatient treatment. The proposed correctional budget sought additional funds to address capacity issues since the prison population is projected to increase slightly next year in a modest reversal of an overall downward trend. More than $18.0 million is slated to go toward a variety of health-related programs providing treatment and benefits to current prisoners. Nearly $2 million would be used to hire 14.5 additional full-time employees with the Colorado State Patrol. The department justified the need for additional highway patrol officers in order to reach their goal of equally dividing officer time between reactive and proactive policing. An additional $1.2 million from marijuana tax revenue will fund a special unit within the Colorado Bureau of Investigation to investigate illegal marijuana trafficking.An updated letter from Governor Hickenlooper to the JBC on March 19, 2018 stated that the revised economic forecasting data indicated an additional $548 million available to for state legislators to spend. The governor’s office proposed allocating nearly half of this amount to transportation needs across the state. Transportation funding has been a source of controversy in recent years, as the legislature appeared to have the votes to refer a modest tax increase to voters for consideration in the prior election year. However, a small group of Republicans in the Senate blocked the measure from progressing out of committee. Transportation funding remains one of the key issues on the legislature’s agenda as the session moves toward a close. As in 2017, Governor Hickenlooper’s final State of the State address called for a transportation tax to be placed on the fall ballot. Last year, bato bucket the General Assembly came close to referring a measure to the voters that would increase the state’s sales tax from 2.9 percent to 3.52 percent. This revenue would be used to secure bonds for local and statewide transportation projects. The referendum passed the House with all Democrats voting in favor in addition to four Republicans. Although it appeared likely that the measure would similarly pass in the Senate, it was defeated in committee. Governor Hickenlooper encouraged lawmakers to revisit the transportation tax increase in the current session by arguing, “We need to be even more ambitious. It’s time we look at a long-term solution with a sustainable funding source. There’s broad agreement — across party lines. Coloradans deserve the opportunity to vote on whether we need new resources and where they should come from. It’s time to go to the voters” . The legislature is currently considering a number of alternatives, but it appears likely that the parties will reach some type of funding agreement soon. The current compromise will likely involve a one-time spending allocation to transportation as the governor proposed along with an agreement to defer any ballot measure to increase taxes for transportation funding to 2019 or beyond . However, it is possible that a citizen’s initiative to raise transportation taxes may nonetheless qualify for the ballot. Coloradans are generally not supportive of such proposals. Voters rejected increased cigarette taxes in 2016, and Coloradans twice opposed income and sales tax increases to fund education in 2011 and 2013. Neither outcome was close. In fact, since TABOR was ratified into the state’s constitution in 1992, no proposed tax increase on the statewide ballot has passed with the exception of an increase in cigarette taxes when the electorate approved Amendment 35 in 2004. Voters did, however, approve the tax levels for the new retail marijuana market in 2013.

If the transportation tax initiative is delayed for at least another year, there will be a different governor in office when voters decide whether the addressing state’s transportation problems merit a tax increase. Based upon the party’s positions in the legislature, one would expect that the Republican candidate for governor would oppose a transportation tax increase, while a Democratic candidate would likely campaign in support of the measure. The 2018–2019 state budget provides substantial financial investments in existing and new programs across a wide array of policy areas. Governor Hickenlooper identified education, transportation, and corrections as among the issues worthy of greater funding. A General Fund budget of $11.42 billion exceeds the budget from the prior fiscal year by nearly 8 percent. This increase in spending is made possible by a confluence of factors including a strong economy, federal tax reform, and state budgetary reform from the prior year affecting how the hospital provider fee pertains to the requirements imposed by TABOR. Yet TABOR remains, as its creators intended, a powerful vehicle to limit government revenue and spending. While the state budget increases funding for nearly every state department, a prosperous economy would normally allow for even greater investments in areas of need. Sustained population growth has expanded the tax base, but the state government remains restricted in the amount of revenue it can collect and spend. Citizen initiatives to increase taxes for education and transportation will appear on the ballot in the 2018 midterm election, but Coloradans have rarely demonstrated a willingness to increase taxes. Accordingly, the constraints imposed by TABOR have curtailed even greater investments in state programs that a strong economy and budgetary reform have helped facilitate.A quick history of ecology might begin with the idea that Tansley’s post-WWI botanical surveys ushered in the modern science of ecology, although its basic vision was articulated half a century earlier by Haeckel, in what was a simple statement recognizable even today in its modern manifestation . The application of mathematics to the field was pioneered by the threesome of Lotka, Volterra, and Gausse, in the 1920s and the conceptualization of an ecosystem as a hierarchical energy transfer system by Lindeman in 1942, arguably the foundation on which the Odum brothers successfully launched what we call ecosystems ecology today . The famous Hairston and colleagues paper, perhaps marking the birth of modern community ecology, generated a popular framework that continues generating interesting research questions, all formulated around the central idea that terrestrial communities are regulated by the predators eating the herbivores. Finally, Richard Levins’s lifelong commitment to viewing both ecological communities and ecosystems as complex systems presaged the current popularity of combining recent developments in the field of complex systems with the persistent search for basic framings in community ecology . This is the historical skeleton on which the current essay is built, framed within the Hairston and colleagues hypothesis in the context of Levins’s insistence on a complex systems approach. The most enduring question of Hairston and colleagues remains popular today, indeed takes on new meaning in the context of modern complex systems. Is a community controlled from above or below, from predators or from competition among herbivores for plant resources ? Certainly ecologists have not been credulous to the point of oversimplifying either predator–prey relationships or interspecific competition; however, the simple question “Is control from above or below?” seems a persistent underlying framework for many research programs. The agroecosystem, by its very nature, refuses to entertain this question, at least in its classical form. If the objects of attention are overused by the herbivores , it is not a system that is regarded as one we should allow to persist . Rather, if the system is to persist at all, the herbivores must be regulated from above. Indeed the standard practical agronomic question “How can we control the pests?” if translated into its true ecological meaning would be “How can we cause the system to switch from control from below to control from above?” Control from below has come to mean much more than seems to have been the intent of Hairston and colleagues , especially in light of what we now understand about plant chemistry and its relationship to herbivory .

In a subsequent 2 mo study of the effects of the nutrient bar on CMR markers in individuals across a range of BMIs, only those with low inflammation at baseline as assessed by high sensitivity C-reactive protein < 14.3 nmol/L responded comparably to those in the earlier trial, with not only increased HDL cholesterol and large HDL particles but also a trend toward increased high molecular weight adiponectin and a decrease in other CMR factors at 2 weeks, sustained at 2 months . In particular, a shift in low density lipoprotein particle subfractions toward a less atherogenic profile was evident in the non-inflamed group . Although the participants with overweight or obesity and CRP > 14.3 nmol/L did not show this response, they did experience an upward trend in adiponectin by 2 months. These results suggest that there may be a continuum of metabolic responsiveness to this nutrient supplement that is slowed in the face of the chronic low-level inflammation commonly observed with obesity and insulin resistance. A 6 month study of this nutrient bar in obese adolescents with non-eosinophilic asthma showed improved lung function at 2 months, but favorable movement in cardiometabolic biomarkers only began to emerge at 6 months.It is not known whether more sensitive biomarkers of early metabolic change may be capable of detecting short-term effects of the nutrient bar supplementation in persons at CMR.

Past metabolomics studies in obese adolescents and adults have identified strong positive associations between baseline levels of branched chain, aromatic, sulfur, and gluconeogenic amino acid metabolites and parameters of inflammation and insulin resistance. Similarly, planting gutter elevated levels of specific ceramide species have been shown to associate with inflammation, dyslipidemia and insulin resistance in both adult and adolescent obesity. The relative sensitivity of these biomarkers to reflect moderate changes in dietary intake of polyphenols, essential lipids, fiber and vitamin/minerals, remains incompletely understood. In the present randomized, controlled, non-blinded trial, a two month intervention with exercise and nutrition counseling alone or with nutrient bar supplementation was performed in a high CMR cohort of adolescent /parent adult caretaker family units to determine 1) cross-sectional relationships in both adolescents and adults between traditional CMR biomarkers and amino acid and ceramide metabolites and 2) longitudinal changes within groups in the same CMR biomarkers following the lifestyle +/- nutrient bar intervention.Anthropometric and clinical evaluation: All anthropometric measures were performed in duplicate in the clinical research center and if not within 10% agreement, were repeated a third time. The reported measure is an average of the two closest numbers. Height was measured with a stationary stadiometer.

Weight was measured using a digital electronic scale and the BMI and waist to height ratio were calculated. Waist circumference was measured at end expiration to the nearest mm with a Gulick II Plus tape midway between the lowest border of the rib cage and the upper border of the iliac crest.Blood pressure and Resting Heart Rate: Each was measured in triplicate after 5 minutes sitting quietly with readings taken at least one minute apart. An automatic digital blood pressure monitor was used with cuff size adjusted for arm size. Traditional CMR biomarkers: Fasting blood samples were drawn and processed in the UCSF Benioff Children’s Hospital Oakland Clinical Research Center and sent to ARUP Diagnostic Laboratories for: standard lipid profile [total triglyceride, total cholesterol, and cholesterol within HDL and LDL , glucose, insulin, 25 hydroxy Vitamin D level and CRP. TG to HDL ratio and non-HDL were calculated. Fasting insulin and glucose were used to calculate the Homeostasis Model Assessment of Insulin Resistance Index according to the formula: fasting insulin x fasting glucose /22.5. Lipoprotein particle sub-classes were analyzed by an ion mobility procedure that sensitively and directly measures concentrations of lipoprotein particle subfractions. High molecular weight adiponectin was measured by solid-phase sandwich ELISA . Metabolomic analyses: 1) Targeted analyses of 42 amine-containing metabolites consisting of 20 major amino acids, and secondary metabolites of arginine and cysteine whose levels are sensitive to inflammation and oxidative stress were performed on stored samples preserved at -70o Fahrenheit. Briefly, plasma was acidified with 5% perchloric acid containing 8 stable isotope internal standards. Acid-soluble supernatant was used for strong-cation exchange solidphase extraction to capture cationic amine- containing metabolites. Extracted metabolites were further derivatized with isopropylchloroformate.

Derivatives of metabolites were resolved using Agilent 1260 ultra-high pressure liquid chromatography and eluted with a gradient of water and isopropanol . An Agilent 6490 triple quadrupole mass spectrometer was used to detect resolved analytes and quantify them using authenticated external and internal standards. 2) Sphingolipidomics by electrospray tandem mass-spectrometry by validated techniques was used to identify sphingolipid metabolites, including ceramides.De-identified baseline and study completion data points, paired by participant study ID, were entered into SPSS . Descriptive analyses of the study cohort were summarized and results for Teens and PACs, as well as for combined CONT and INT groups, were compared by unpaired Student t-test. Continuous physical and metabolic variables were tested for normality by examining the skewness, kurtosis and the Shapiro Wilk tests and transformed as necessary before analysis. Most of the variables in our data were normally distributed. Log transformations were conducted for the continuous physical and metabolic variables that were skewed to make them as normal as possible. The Shapiro-Wilk tests show that all of the transformed variables except two are approximately normal. The measures of metabolite concentrations used for principal component analysis were Z-transformed to render them normally distributed on the same scale with mean of zero and standard deviation of one. Pre-post change in absolute metabolite levels were compared by repeated samples paired t-test. Baseline z-scores of metabolites were subjected to principal component analysis without rotation. PCA is an unsupervised analysis that aims to decrease the complexity of data by reducing variables to a smaller number of principal components . Each of the PCs were vectors of metabolite contributions. A direct oblimin rotation was used and 6 factors before the bend in the scree plot , and eigenvalues >1 were retained. Component scores for each participant were calculated with a standardized scoring coefficient. A PCA model with oblique rotation was tested to examine the factor correlation matrix. Since none of the factor correlations were over ± 0.32, indicating factor correlations are not driven by the data, we estimated a model with orthogonal rotation to reveal a simplified structure with interpretable factor loadings. To identify metabolite patterns of interest, bivariate Pearson-correlation analysis was performed between percent changes in metabolites and clinical biomarkers of CMR and with lipoprotein particle subfraction distributions separately. A generalized estimating equation procedure determined the significance of longitudinal changes in PC scores and their contributing metabolites using age and gender as co-variates. Wald Chi-square tests determined the significance of pairwise differences in treatment responses. Statistical analyses were performed using IBM SPSS Statistics version 25 and Rstudio version 1.1.456, and p-values <0.05 were considered statistically significant for the differences in traditional biomarkers. A Bonferroni correction for multiple comparisons adjusted the p-value set as significant to <0.002 for the correlational analyses with amino acid and ceramide metabolites.The study cohort was predominantly female, 61% Hispanic , 25% Nonhispanic Black, 8% Hispanic Black. and 6% Nonhispanic White, gutter berries with more Hispanic Not Black than Nonhispanic Black participants randomized by chance to the INT group . Full assessment of physical, behavioral, metabolic, and metabolomic status was conducted at baseline and study completion on the cohort of 36 participants . There was excellent attendance in both INT and CONT groups with all Teens and PAC participating in more than 78% of group sessions and 100% of baseline and follow-up assessment visits. There was considerable prevalence of obesity, dyslipidemia, inflammation and insulin resistance in all participants and mild hypertension in adult participants at baseline . Mean baseline BMI was over 30 mg/m2 in all participants, did not differ by sex, but was significantly higher in the CONT group relative to INT. The mean CRP was over 28.6 nmol/L in all participants but higher in Teen participants assigned to the CONT group than to the Teen INT group, and between the combined CONT and INT groups. Despite these random differences in BMI and CRP, central adiposity , blood pressure, lipids, insulin resistance and inflammation were similar in both groups, suggesting closely matched CMR profiles.

Although in population studies, Waist Circ is greater in males as compared with females, only one of the 18 PAC was male; while his Waist Circ fell below the adult female PAC mean, it was not the lowest recorded among PAC. The average Waist Circ for Teen males vs female Teens did not differ significantly so Waist Circ data are not reported by sex. Compliance with nutrient bar intake by self-report was 85.8 ± 11.1% and 86.7 ± 13.8% among INT group adults and Teens respectively.The quality of self-report diets assessed by Block food frequency questionnaire was poor in all participants at baseline . Modest but significant improvement in CONT PAC was evident in categories of saturated fat intake and added sugar but trends in the same direction did not reach significance in INT PAC or in Teens from either group. Average daily servings of dietary fruit , vegetables, not including potatoes , and fiber were comparably low in both CONT and INT groups and did not change in any age subgroup . Self-report activity increased in all participants in both groups . Weight was stable, even in the INT group despite the addition of 220 kcal in two daily nutrient bars. Good compliance with nutrient bar intake was suggested by a significant increase in plasma 25 hydroxy Vitamin D in the INT group PAC and Teens, but neither CONT subgroup. There were no other consistent effects of nutrient bar supplementation evident in traditional CMR biomarkers, nor were there significant within- or between group changes in any anthropometric measures in combined CONT and INT groups. Subgroup analysis of PAC and Teens separately showed some favorable changes in both CONT and INT groups. PAC CONT participants decreased Waist Circ . Among Teen participants, CRP trended lower in CONT but did not reach significance . Among Teens, systolic blood pressure decreased significantly by 7% in INT only, trending upward in CONT Teens with a significant between-group difference.Previous bar supplementation trials showed that chronic inflammation was associated with slower response in metabolic improvement. All participants, both Teens and PACs in this study, met criteria for obesity and had baseline CRP > 14.3 nmol/L.To test the hypothesis that subtle shifts in metabolism may precede changes in traditional biomarkers, a targeted analysis of plasma ceramides and amino acids was performed. The top six PCs extracted, each composed of distinct sets of linked metabolites, explained 59.3% of the total variance in the data set . PC1, which explained 16.7% of the variance, was composed exclusively of ceramides and ceramide-1-phophates; PC2 of amino acid metabolites; PC3 of sphingomyelins; PC4 of dihydroceramide species, glucosylceramide, and one specific ceramide subspecies Cer 18:1; PC5 of biomarkers of meat consumption and transsulfuration and polyamine metabolites ; and PC6 of amino acids in antioxidant defense .We next determined how increased physical activity by itself or with nutrient bar supplementation modulated these baseline metabolomic parameters. Analysis of PC change was performed with adjustments of potential covariates including age, gender, and baseline CRP. While there was no change in the plasma total ceramide pool, results showed significant divergence in several key ceramide species within PC1 between CONT and INT. Results show significant increases in C14:0 , C20:0 , C22:0 and C24:1 ceramides and a nearly significant increase in C16:0 in CONT. In contrast, C24:1 ceramide significantly decreased in the INT group by 18%; all other ceramides trended lower or remained unchanged . Analysis of group by time interactions showed significant between group differences in all of these C14:0, C16:0, C20:0, C22:0 and C24:1 ceramide species. Sphinganine and sphingosine are essential substrates for ceramide synthesis through de novo synthesis and salvage pathways. In CONT only, levels of sphinganine and sphingosine increased significantly by 59% and 50%, respectively resulting in significant pairwise differences between CONT and INT groups .Sphingosine-1-phosphate is a terminal breakdown product of ceramide and an important anti-inflammatory and vascular signaling lipid mediator associated with lipoproteins, particularly HDL, in plasma. As shown in Table 5, S1P levels increased differentially in both CONT and more so INT participants. The S1P increase in the INT group differed from the degree of change in CONT at p = 0.007. Table 6 lists amino acid metabolites with significant within group and between group changes.

There is no evidence of GLRaV-3 transovarial passage . While laboratory-based transmission studies have been informative, there is still a need for field-based transmission experiments to accurately estimate expected vector transmission efficiency and disease incubation time within the host. Information is lacking regarding how soon disease symptoms will appear after vector-borne infections are initiated, or how soon crop quality will be affected. The goal of our study was to obtain information about vector-borne transmission of GLRaV-3 and subsequent disease progression under commercial vineyard conditions. We performed a controlled GLRaV-3 transmission study into mature V. vinifera cv. Cabernet franc vines in Napa Valley, CA USA. We used first instars of the vector Pseudococcus maritimus , a mealybug species that is a common vineyard pest and native to North America . We performed a concurrent laboratory study, with identical experimental design, to compare pathogen transmission efficiency under controlled laboratory and field conditions. In the field study, dutch buckets system we estimated transmission efficiency and monitored time to GLRaV-3 detection via molecular diagnostics, appearance of symptoms, and effects of disease on berry quality.

Our findings are key to informing sound management practices with respect to understanding spread and progression of disease in a commercial vineyard. Furthermore, we provide a previously missing link between controlled laboratory studies and realistic vineyard conditions.Virus-infected dormant cuttings of V. vinifera cv Cabernet Sauvignon were used as source of GLRaV-3 in our transmission experiments. Foundation Plant Services at the University of Califtornia, Davis provided accession LV89-01 from their Virus Source Vineyard, which is known to be infected with genetic variant group III of GLRaV-3, Grapevine virus B , and Grapevine fleck virus . This accession was chosen because genetic variant group III of GLRaV-3 is common in Napa Valley . Plant cuttings were cut to three buds each, treated with RootBoost rooting hormone, planted in 1:1 vermiculite: perlite, and kept on a mist bench for 6 weeks, until a few leaves were produced and roots were approximately 2.5 cm long. Cuttings were then removed from the mist bench and transplanted to 10 cm pots with a growth medium consisting of 2:1:1 SuperSoil: perlite: sand , and kept in the greenhouse until used as virus sources in transmission experiments. GLRaV-3 infection was confirmed by molecular diagnostics prior to use for source material in transmission studies. Virus-free dormant V. vinifera cvPinot noir cuttings were also provided by Foundation Plant Services, collected during winter dormancy 2011, and propagated in the same manner as the virus-infected source cuttings. We used Ps. maritimus as the mealybug vector, which is difficult to maintain in insect colonies; therefore we relied on field collections for experimental inoculations. To obtain virus-free first instar Ps. maritimus mealybugs, third instar females were collected from a vineyard in Pope Valley, CA in May 2011, and allowed to mature and oviposit in the laboratory.

The third instar females were collected from underneath the bark of the trunks and cordons of mature grapevines and placed into gel capsules for transport to the laboratory. The mealybugs were immediately transferred to 100 mm petri dishes, each containing one piece of 70 cm Whatman filter paper. The mealybugs were kept in darkened conditions at 25 °C: 20 °C, 16: 8 h day: night temperatures- females were removed and discarded after oviposition. The filter papers containing ovisacs were pinned to the underside of the aforementioned infected source plants, which were then kept in a growth chamber until the first instar mealybug crawlers hatched. Approximately 72 h after hatching on the infected source plants, mealybugs were transferred to mature vines in the vineyard and to uninfected vines in the laboratory, for a 48 h inoculation access period. The timing of hatching led us to perform field inoculations on 18 July 2011, which coincided with the emergence of the new Ps. maritimus generation in Napa Valley. Twenty replicate source vines were propagated and used, with one to five recipient test vines inoculated per source plant in each inoculation experiment . All recipient test vines were treated with an insecticide upon completion of the inoculation access period.The experimental field inoculations were located in three rows of a vineyard block of V. vinifera cv. Cabernet Franc clone 01 grafted to 110R rootstock, obtained from Duarte Nursery and planted in Oakville, Napa Valley, CA in 1994. No vines in the experimental area were symptomatic for grapevine leaf roll disease prior to our experimental inoculations.

To confirm initial GLRaV-3-free status prior to inoculations, three petioles were collected from each experimental vine in July 2011 before inoculations were performed, for diagnostic testing . The block consisted of 8315 vines planted at 588 vines per hectare. Row spacing was 1.8 m, and vine spacing was 1.5 m, with a vertical shoot positioning trellis system and bilateral pruning. Row direction was northwest-southeast. Drip irrigation was provided using one 3.8 – L·h–1 emitter every 1.5 m. Experimental vines were spaced every third vine, and treatments were fully randomized. The three treatments included inoculations with no leaf cages, inoculations using mesh leaf cages, and negative controls for which no experimental manipulation was performed. Each treatment included 30 replicate vines, for a total of 90 experimental vines. The experiment comprised an area including 360 total vines, including the 90 experimental vines plus the spacer vines. The spacer vines were monitored periodically throughout the study for symptoms of grapevine leaf roll disease. A survey for any signs of mealybugs was performed in October 2012. On 11 October 2012, 15 months post inoculation, a commercial testing service collected and analyzed material from some vines that were symptomatic for grapevine leaf roll disease in the experiment and tested for a broad panel of known grape pathogens: GLRaV-1, GLRaV-2, GLRaV-2 strain Red Globe, GLRaV-3, GLRaV-4, GLRaV-4 strain 5, GLRaV-4 strain 6, GLRaV-4 strain 9, GLRaV-7, Syrah virus 1, Grapevine virus A, GVB, Grapevine virus D, Grapevine fanleaf virus, Xylella fastidiosa, GFkV, Rupestris stem pitting-associated virus, Rupestris stem pitting-associated virus strain Syrah, and Grapevine red blotch-associated virus. The vineyard block was not treated with insecticide prior to inoculations during the 2011 growing season. For inoculations, ten Ps. maritimus first instar insects were gently moved with a paintbrush from leaves of infected source plants onto the underside of one fully expanded mid-height leaf, located on a vertical cane growing from a middle spur on the south cordon of each grapevine. For the caged treatment, a cloth mesh cage was placed over the inoculated leaf and secured at the petiole using a twist tie. For the uncaged treatment, no covering was used on the inoculated vine. The experimental area was commercially treated with spirotetramat insecticide on 20 July 2011, after a 48 h inoculation access period. After inoculations the experimental area was managed following standard commercial practices.

Three months after inoculations, dutch buckets the petiole of the inoculated leaf was collected on 14 October 2011 for diagnostic testing. In the instance where that petiole had fallen off the vine or could not be found, a petiole near the inoculated leaf was collected; inoculated petioles were missing from 9 of 60 inoculated vines. Immediately following the first appearance of symptoms in 2012 and 2013, petioles were collected from each experimental vine and tested for presence of GLRaV-3. Petioles were collected from each experimental vine in September 2014, and tested for the presence of GLRaV-3, GVB, and GFkV. On each sampling date, three petioles were collected from each vine and pooled for diagnostic testing. If a vine had symptomatic leaves at the time of sample collection, symptomatic leaves were preferentially collected over asymptomatic leaves. During each growing season in 2011 through 2014 , experimental vines were surveyed regularly for visible leafroll disease symptoms, beginning immediately after inoculations. On each survey date vines were marked as either asymptomatic or symptomatic, with surveys beginning in May and continuing through October. Shortly after symptoms first emerged in 2012, a detailed symptom survey of each symptomatic vine was performed to determine possible variation in disease symptom severity among vines and if there was an association between location of inoculation and initial appearance of symptoms within vines. For this survey, the position of each spur and the number of symptomatic and asymptomatic leaves on each spur were recorded. In Year Two, berry quality of all vines was measured three times during the weeks immediately preceding commercial harvest. Degrees Brix , pH, and titratable acidity were measured on 31 August, 21 September, and 3 October 2012, and harvest was 4 October 2012. In Year Three, berry quality of a randomly selected subset of 30 vines was measured on 28 August and 14 September, and harvest was 14 September 2013. The 30 vines were evenly divided between uninfected negative controls, uninfected and infected vines from the caged inoculation treatment, and uninfected and infected vines from the uncaged inoculated treatment. For berry quality analysis, on each sampling date approximately 200 berries were collected from each vine to minimize variance in measurements . Within each grapevine, berries were collected from the top, middle, and bottom of each harvestable cluster of grapes and pooled for laboratory analysis. All samples were processed by Constellation Laboratories in Califtornia, USA. Total soluble solids as °Brix were measured using an Atago refractometer, and pH was measured using an Orion pH meter. Titratable acidity of the juice was measured via direct titration with 0.1 N NaOH, using phenolphthalein as an indicator.To compare field inoculations in an established commercial vineyard with controlled laboratory studies, a laboratory experiment including the same treatments was performed, using the same collection of Ps. maritimus and same known infected source plants. Ps. maritimus first instars were moved from known infected source plants to uninfected recipient test plants on 15 July 2011 and allowed a 48 h inoculation access period, after which plants were treated with a contact insecticide. Mealybugs on half of the recipient test plants were caged on the underside of one leaf using clip cages, as described by Tsai et al. , and mealybugs remained uncaged on the other half of the recipient test plants. Thirty replicate recipient test plants were used for each treatment, caged and uncaged, for a total of 60 recipient test plants. Additionally, a replicate of 30 uninoculated plants was kept in the greenhouse following inoculations to assure that all GLRaV-3 infections were due to our experimental inoculations. Following inoculations, plants were kept in a greenhouse with 16:8 light: dark using supplemental lighting as needed, and 25 °C: 22 °C day: night temperatures, with regular fertilizer and insecticide treatments. Because disease symptoms under greenhouse conditions are not reliable for diagnosis of GLRaV-3, we used molecular testing to confirm infection with GLRaV-3. Four months after inoculations, at which time new GLRaV-3 infections can be reliably detected using molecular diagnostics, three petioles per inoculated test plant were collected and stored at −80 °C until testing for GLRaV-3 .To test whether the newly infected field vines could be a source of GLRaV-3 one season after mealybug inoculations, a transmission experiment was performed in the laboratory from cuttings of these newly infected fieldvines. Ps. maritimus were not used because of the abovementioned difficulty in obtaining virus-free first instars for transmission experiments. Instead we used first instars of Planococcus ficus, which are easily maintained in colonies and therefore can be ready for use in transmission studies at any time. Furthermore, Pl. ficus is a known vector of GLRaV-3 . Field cuttings were collected on 4 October 2012 and the stem bases were placed in flasks of water. First instar Pl. ficus were allowed a 24 h acquisition access period on the field cuttings, then transferred to the underside of a leaf of virus- free V. vinifera cv. Pinot noir recipient test vines; ten insects per recipient test vine were confined using a leaf cage for a 24 h inoculation access period. Following inoculations, plants were treated with a contact insecticide and then kept in a greenhouse for four months until petiole sample collection for diagnostic detection of GLRaV-3. For this experiment, a randomly selected subset of experimental field vines of each treatment was tested as a potential GLRaV-3 source. In total, nine symptomatic vines were tested; five from the caged inoculation treatment and four from the open inoculation treatment, and seven recipient test vines were inoculated in the laboratory from each symptomatic field vine. One of these 63 recipient test vines died before petiole sample collection to test for infection with GLRaV-3.

Our modification accounts for the effect of temperature on the survival and fertility of D. suzukii in calculating population growth of the organism. Typically, researchers have introduced elements of environmental stochasticity to matrix models to study environmental effects on population trajectories. However, our approach relies on temperature-dependent estimations of age specific fecundities and survival that are determined by models fit to lift table data generated for multiple temperatures. Our environmentally dependent matrix model is unique in that it does not rely on simulation of environmental effects on populations, but the matrix itself is recalculated at each iteration in direct response to environmental input. Model predictions were run under environmental conditions from different regions to illustrate variation between and within study sites in different years. These simulations make important predictions about age structure and population trends that have implications for pest management both in a broad sense and with regional specificity. This modeling tool may improve current management practices by predicting pest pressure independent of trap catches or samples of infested fruit. We also see potential applications of this model for research in other fields of study and for broadening the understanding of how pests interact with the environment.

The population projection model was written in the open source statistical environment R version 3.0.2. The model calculated the matrix based on mean temperature input. Briefly, nft hydroponic the matrix calculations were based on age-specific regressions of temperature-dependent population parameters as highlighted by Tochen et al.. Whereas immature lift stages of D. suzukii may experience different environmental conditions than adults because these lift stages are completed within the fruit, in this study, ambient air temperatures were used to predict population dynamics for all lift stages. To return age-specific population vectors for 50 age-classes of D. suzukii for each test case, a vector of mean daily temperatures for each site was input into the R statistical interface. The biofix, or the point where the model began in the spring, was determined using methods described in Tochen et al.. Biofix essentially described the earliest point in the season when the temperature allows the population to increase. Calculations for population estimates were initiated on the biofix date of 2 February in Parlier and 1 April in Wilmington and Salem . In Pergine and Sant’Orsola, estimates were initiated on 6 April . The population matrices were initiated with 100 flies in the population vector for 41–50 day-old females based on the assumption that females of this age group represent flies that would be emerging from diapause in spring. The log transformed sum of D. suzukii from all lifte stages for each day represented the total population estimate except where age distributions are considered. For daily age distribution of D. suzukii from Parlier, Salem and Wilmington during 2013, 1–3 day-old D. suzukii were classified as eggs, 4–7 day-olds were larvae, 8–9 day-olds were pupae, and 10–50 day-olds were classified as adults.

Among the most important assumptions of the model are that populations of D. suzukii would not be limited by host availability, are not density dependent, do not exhibit Allee effects, and that response to current temperature is not dependent on previous temperature exposure.Seasonal weekly trap catches of D. suzukii were recorded in all study sites, except Riva del Garda, but model estimations for this location was included because the climate here is much different from the other locations studied in Italy. Trap counts were pooled data from commercial blueberry fields in Wilmington ; unsprayed apricot, blackberry, blueberry, cherry, peach, and citrus orchards in Parlier ; commercial blueberry fields and surrounding blackberry vegetation in Salem ; strawberry, blackberry, cherry and blueberry fields in Pergine ; and unsprayed strawberry and raspberry fields in Sant’Orsola . In Wilmington and Salem, traps were made of clear plastic cups, ca. 1 liter in volume each. Each trap had 6–15 entrance holes 4.5– 9 mm in diameter. Trap baits in Wilmington consisted of a yeast and sugar water mixture containing 6 g yeast and 40 g sugar dissolved in 710 ml water. In Salem, traps were baited with 100– 200 ml natural apple cider vinegar and 1–3 ml unscented liquid soap to break water surface tension. In Parlier the traps were made to the specifications of the ‘‘Haviland Trap’’ design for D. suzukii monitoring. A 750-ml plastic container served as the basin for each trap. A 7.5-cm diameter hole was cut in the lid, over which a piece of 0.6-cm wire mesh was attached. Each trap was covered with a Pherocon trap cover , which had a built-in wire hanger. Each trap was filled with 250– 300 ml of apple cider vinegar with 15 ml of unscented soap added as a surfactant to each container of vinegar. In Trentino the containers were 1000-ml graduated white polyethylene bottles filled with 200 ml apple cider vinegar . All traps were placed near the fruiting level of host plants or on stable surfaces in shaded areas and were checked weekly.

The contents of each trap were collected into a separate container that was taken to the laboratory for processing, and at the same time, the traps were refilled with fresh apple cider vinegar and unscented soap, as described above, in the field. The liquid and contents from each trap sample were strained in the laboratory and the numbers of adult SWD collected were recorded by gender. All data from traps were analyzed to display mean weekly D. suzukii per trap for each of the regions. Mean daily temperatures for all seasons are presented together with trap catches.In Wilmington, winter and late dormant mean temperatures were never below 0uC and reached 20uC multiple times prior to 31 April . At this site, mean high summer temperatures exceeded 25uC multiple times during July, and mean low temperatures were below 10uC during November. In Parlier, winter and late dormant temperatures were never below 0uC and were higher than observed in both Salem and Wilmington during this period . Here temperatures were above 25uC multiple times prior to 31 April. Mean daily temperatures were consistently above 25uC during June–September, and daily means in Parlier sometimes exceeded 33uC. In Parlier, temperatures dropped to below 10uC during November. In Salem, early-season temperatures were warmer during January and February of 2012, compared to 2013 . Temperatures observed from March through May were slightly higher during 2013 compared to 2012. In Salem, winter and late dormant temperatures were never below 0uC or above 15.3uC until 31 April 2013 . Daily mean temperatures gradually increased to 25uC during July, after which daily mean temperatures dropped to below 10uC during November. For the Italian sites Riva del Garda had higher late dormant and fall temperatures than Pergine and then by Sant’Orsola . Summer temperatures were similarly ranked higher in Riva del Garda, followed by Pergine and then Sant’Orsola during 2013. In Pergine during winter 2012 , late dormant temperatures were frequently below 0uC and were as high as 20uC before 31 April. Fluctuations of temperature were more pronounced in 2012 in Pergine, compared to 2013 . Very low temperatures were recorded in Pergine from 3–12 February 2012, followed by relatively warm temperatures from 24 February to 13 March. Temperatures were also comparatively low from 9–11 April 2012. In Pergine, daily mean temperatures increased to 25uC during July, after which they dropped to below 10uC during November. Mean temperatures were well below 0uC during December.Population estimates using temperature data indicate that D. suzukii populations are able to increase to high levels in all of the studied locations . When comparing early-season population estimates between Wilmington, Parlier, and Salem , hydroponic gutter the population estimates were highest in 2013 in Wilmington followed by Parlier and then Salem. However, the population estimate for Salem surpassed Wilmington by 15 June and surpassed Parlier on 16 July, as Salem population estimates continued to climb while the latter sites experienced declining populations after reaching the first peak of their bimodal distributions. In Parlier, the early-season population peaked on 16 June, subsequently decreasing to a low on 10 September before increasing to a second population peak on 9 November, then decreasing again as winter progressed.

In Wilmington, the population curve peaked on 21 June, then the population curve declined slightly for an extended period, followed by a second period of population increase beginning on 19 September to a population peak in November. In Salem, populations consistently increased from 25 April to a peak on 22 October, followed by a steep decrease. When comparing population estimates between seasons for the initial harvest period of early- to mid-season blueberries in Salem , the majority of model outputs for this period estimated greater populations for 2013. When comparing populations along the elevation gradient of the three Italian sites, higher early-season populations were predicted at the lowest elevation Riva del Garda, followed by Pergine and then Sant’Orsola . In Pergine, greater population numbers were estimated for the majority of the growing season during 2013 compared to 2012 . In all model predictions, immature lift stages comprised by far the majority of the population, except at the beginning or end of the season when adults tended to dominate . One exception was Wilmington, where temperatures remained favorable for reproduction into the late fall so that immature stages remained a majority of the population . In Salem, fall temperatures initially caused cessation of reproduction, leaving a majority of adults, but December temperatures allowed for some reproductive activity to occur . In the early spring, a higher relative percentage of adults occurred due to the overwintering adults that were initiating their first reproduction. In part, this was an artifact of initiating the model with only older adult females. In the fall, environmental conditions became unfavorable for reproduction but may not have had strong effects on adult survival. Overall, no populations reached a completely stable age structure, but the highest relative stability for each site occurred in the middle of the season. Stability of age structure was the highest in Wilmington, followed by Parlier and finally Salem, which had a high degree of instability . Demarcation of distinct generations was very clear for the first part of the season in Parlier and Salem , but during the mid season at these sites and in Wilmington , it was very difficult to distinguish individual generations to distinguish complete generations from partial generations.In Wilmington, D. suzukii counts were first recorded on 5 May 2013 at one fly per trap with an erratic increase to a peak in numbers at 26 flies per trap on 26 July . After this period, the trap numbers gradually decreased to six flies per trap until 4 December, at which point D. suzukii trapping was discontinued. In Parlier, two population peaks were found during the crop season, one during the early part of the season, followed by a long mid-summer period without fly captures, and a second peak during the latter portion of the season . Adult D. suzukii were first caught on 19 March 2013 at one fly per trap and increased to a high of six flies per trap on 16 May, after which they decreased to zero on 27 July. The trap numbers remained at this level until 19 September, after which numbers continued to increase into December. In Salem and Wilmington only one population peak was observed during the summer period . During 2012 in Salem , D. suzukii trap counts consistently increased starting on 5 July from one fly per trap per week to a maximum average of 17 flies per trap on 6September. During 2013 in Salem , the first D. suzukii trap counts were observed on 30 May at an average of three flies per trap per week and gradually increased until 10 September, when a maximum of 27 flies per trap was observed. The first trap counts during 2013 were therefore consistently recorded four weeks before those found in 2012 and higher levels of flies were found in traps during 2013 in Salem. In Pergine and Sant’Orsola, fly counts were first observed 23 June 2013 , and on 7 July 2013 in respectively. In Italy, one population peak was visible each year for Pergine and Sant’Orsola. In 2012 in Pergine, the first flies were trapped on 25 July, approximately four weeks before those caught during 2013.

Direct competition occurs only among the betilids species, while indirect competition occurs with P. coffea. Gutierrez et al. considered symmetric competition among the betilids, but new data enables parametrization of asymmetric competition in dyadic contests the success rate of which depends on their biology . The different success rates are included in the model via the parameter ı , which weights the proportion of contest that are successful for the dif-ferent combinations of species. No information is available on rare triadic interactions, and hence is not considered in our analysis. When a second species of a betilid parasitoid enters a berry previously colonized by another parasitoid species, it attacks the immature stages of both CBB and the first parasitoid. Facultative oviposition and host-feeding interactions between betilid species are summarized in Table 4 .Simulations for Colombia had a five year time horizon corresponding to the normal crop cycle . In Brazil, the crop cycle is four years. In Colombia, two coffee harvests occur per year, a main one in September-November and a minor harvest in April-June . At Londrina PR , plastic flower pots only one coffee harvest occurs per year that may occur during June through September .

The ratio of parasitoid released was 1 per 1 CBB infested berry. In Colombia, the first berries colonized by CBB females appear around 5300 dd after planting , and at 7500 dd in Brazil. The times of berries first appear are reference times for parasitoid release. In Colombia, an initial release of 11 adults/plant of each betilid species occurs at 6790 dd and corresponds to a ratio of 1:1 parasitoid adults to CBB infested berries. For P. coffea, the initial release was 5 adults/plant corresponding to a ratio of 1:1 parasitoid females to CBB infested berries at time 5678 dd. For Brazil, the initial release of each of the betilid parasitoids at 8500 dd is 4 adults/plant corresponding to a 1:1 ratio of adult females of each betilid species to CBB infested berries. For P. coffea, 6 adults were released at 7500 dd for a 1:1 ratio of parasitoids to CBB infested berries. Single and multiple parasitoid releases during the crop cycle were also evaluated. Two strategies for multiple releases were evaluated: the first consisted of one release at the start of each coffee production cycle when CBB attack begins and the second in had releases at the start and end of each production cycle. The release rates were one female parasitoid per infested berry. The season long damage reduction for each scenario is computed as the ratio of CBB infested berries with control/CBB infested berries without control, while the proportion of parasitoid colonization rate by each species was computed as the ratio of CBB infested berries with parasitoid/total CBB infested berries. A constant immigration rate of 0.25 CBB females per day and plant was used in the model, while no immigration was assumed for the parasitoids. Dyadic contests, facultative host-feeding and facultative hyperparasiThism as appropriate for each species were examined for all combinations of parasitoid species interactions .

The effects of interspecific competition were analyzed using the number of adult parasitoids of each species produced per plant as affected by the presence of competing parasitoid species.The effects of weather at five locations on the coffee agroecosystem were analyzed for the 24 = 16 combinations of the four parasitoids including the null set. The different combinations in the model were characterized using Boolean presence , absence values. The efficacy of each parasitoid species on CBB control was further evaluated using permutations of the years of weather data from the five locations obtained using the allPerms procedure of the “permute” package in the statistical software R . For Colombia, this resulted in a random subset of 24 combinations of yearly weather data and 16 combinations of parasitoids . A similar procedure was used to generate 128 scenarios for Brazil.Negative binomial mixed regression models were fit to simulation outputs. This method accounts for over dispersion and skewness in the data, saThisfies the assumptions of the parametric analysis, and allows interpretation of the stochastic effect of the combinations of years of weather obtained by the shuffling procedure. In the analysis, the Boolean variables are the independent variables, while the cumulative number of CBB infested berries·year−1 in the simulation runs are the dependent variables. The effects of species-specific biology such as facultative host-feeding, facultative hyperparasiThism and dyadic contests were critical underlying factors affecting the output of the model .

The simulation runs were used to estimate the effects of parasitoid species on CBB infestation levels and oneachother.Akaike’s information criteria was used to select the final model for the different combinations of parasitoids, and chi-square goodness of fittests were used to assess the significance level.The dynamics of CBB infested berries in the absence of the parasitoids are depicted in Fig. 2a and predict CBB infestation of berries of 64.67%. Because parasitoid releases at the beginning and at the end of each harvest period for each species acting alone gave only slightly better control than the single release strategy , only the results for single releases are illustrated in Fig. 2. A single release of C. stephanoderis at 10 CBB infested berries/plant resulted in a 21.06% reduction of CBB infested berries on the plant and an 11.35% reduction in infestetation in shed berries on the ground. Compared to C. stephanoderis, a single release of C. hyalinipennis resulted in a 16.93% reduction in infestation on the plant and 8.23% reduction in infestation in shed berries, whilst a release of P. nasuta yielded 16.83% and 7.59% reductions respectively. In contrast, a single release of the eulophid P. coffea resulted in a 47.38% reduction in CBB infested berries on the plant and a 46.85% reduction in infestation in shed berries. Operating singly, the order of effectiveness of the parasitoids is P. coffea >> C. stephanoderis > C. hyalinipennis = P. nasuta , but none of the parasitoids provided high levels of control. Simulation of CBB infested berries available for attack by the betilid species shows asynchrony between seeking adult parasitoids and the low availability of CBB infested berries . The low level of control by single releases of the betilids is the consequence of the lag between the peaks of colonization of CBB infested berries attacked by the betilids and the late emergence of searching betilid females at the end of the ripening season . of Regressions of CBB infested berries·year−1 on the presence the parasitoid species under multiple climatic conditions shows that only P. coffea has a significant impact in reducing CBB infestation levels , a level of control that is clearly insufficient.From a human perspective, control of coffee berry borer using effective augmentative and/or classical biological control is desirable, but this outcome appears not to be possible in Central and South America using the three commonly introduced parasitoids . Of these parasitoids, only P. nasuta established in Colombia , and only C. stephanoderis and P. nasuta established in Brazil . A fourth parasitoid, C. hyalinipennis was considered for introductions, but laboratory studies revealed it is a facultative predator and hyperparasite on immature stages of the other betilids and conspecifics, plastic garden container it super parasiThism of CBB larvae, and it does not attack CBB adults . Our simulation results confirm the impact of these detrimental effects.We sought to determine why the establishment of the parasitoid species in the face of high CBB infestations in Colombia and Brazil has been difficult, and why no combination of the parasitoids shows promise for effective control of CBB. We first review the simulation results and then relate them to ecological theory. In sharp contrast to what is observed in the field, we assumed that all of the parasitoids established in the field and are attacking CBB and interacting with each other. This discrepancy with the facts suggests deficiencies in our knowledge of the biology of the species, and the omission in the model of the limiting effects of harvesting, crop management and cultural controls, and alternate hosts. For these reasons, our analysis of the parasitoids for control of CBB must be viewed as heuristic.

Earlier simulation results by Gutierrez et al. and here suggest that only the eulophid adult endo-parasitoid can reduce CBB populations significantly but not sufficiently. Unlike the betilids, P. coffea attacks multiple CBB females as they are boring into coffee berries and effectively stops CBB reproduction. Furthermore, P. coffea does not host-feed on siblings, allowing rapid growth of its populations during the berry-ripening period. Studies with P. coffea in Costa Rica and Mexico using sleeve and field cages support our simulation results. In Costa Rica, Espinoza et al. released a 1:10 ratio of P. coffea adults to CBB infested berries in entomological sleeves and obtained 95% parasitization of CBB adults and a 3 − 5.6 fold decrease in berry damage. In a similar study in México, Infante et al. obtained P. coffea parasiThism rates of 79%. Unfortunately, cage results cannot be compared with open field releases such as those of Castillo et al. who obtained a very low progeny from the original number of female adults liberated. In a two year survey of CBB parasitoids in Western Kenya, only 17 P. coffea adults were reared from 69,500 CBB infestedberries . These differences in the field studies suggest that crop management, harvest intensity, weather and other unexplored factors must play large roles that will be explored in a forthcoming report.Gutierrez et al. found that the betilids released singly or in combination assuming high immigration and search rates gave poor control of CBB. They attributed the result to a low numerical parasitoid response relative to CBB as each betilid female colonizes only one berry and only partial mortality of the available hosts may occur. Our results confirm this interpretation. Further, the oviposition rate of betilids estimated from field data ranged from 0.055 to 0.076 eggs·female−1·dd−1 which is about six fold lower than the oviposition rate of 0.348 eggs CBB female−1·dd−1 . In addition, host-feeding by betilid adults on CBB eggs and larvae is common and reduces the future availability of preferred stages for parasiThism. Under laboratory conditions, C. stephanoderis females fed honey had 80% greater longevity and decreased host feeding, and this practice has been proposed to improve the survival in the field after release and to improve control of CBB, especially when preys are scarce . In Colombia, high densities of betilid females emerge at the end ofthe berry ripening seasons when CBB infested berries are in short supply. Thus, despite predicted high parasitization rates, a bottleneck occurs between harvests that reduces adult betilid densities to levels that are slow to rebuild in the face of rapidly increasing CBB populations. This result was verified by Benassi in the states of Espiritu Santo and São Paulo, Brazil who observed that low densities of wasps at the beginning of the harvest period required several months to increase significantly. Benavides et al. suggested that parasitoid releases should be made during the middle of the ripening season, but high CBB damage rates normally occur by this time.Whatever strategy a species evolves, it must assure its continuity in time, and hence its traits must assure “adaptedness” to the biotic and abiotic conditions to minimize the likelihood of extinction . Further, Gutierrez and Regev proposed that evolved strategies may have little to do with maximizing the commonness of the species in the environments. Rather, adapted individuals and the species they comprise must via their genetics, take a long-term view of their resource management problem, and that tradeoffs are based on sufficiency or constrained optimization behavior rather than maximization rules. On the theoretical r-K continuum , coffee and CBB have r-selected strategies, while the parasitoids have intermediate ones. The well-documented biology of coffee, CBB and its parasitoids suggests that poor control of CBB is not unexpected because coffee is a long-lived shrub that produces high numbers of fruit, and few seeds need survive CBB attack to provide replacements . Furthermore, CBB is polyphagous in its home range in Africa with coffee being but one of its hosts . Similarly, the linkages of the parasitoids to CBB are not particularly tight because the parasitoids also have other hosts and CBB and the parasitoids spread the risk of extinction through polyphagy . As a result, little by way of regulation of CBB occurs resulting in high infestation levels in coffee in the Americas and in its native range in Africa.

A reliable interpretation of date consumption based on the number of seeds can also be hampered by the possibility of trading crushed dates. Such dates might have been destoned, as could be demonstrated by the Kellis agricultural account book, which mentions a price of 2.5 times that of ordinary dates . Crushed dates with their seeds still present were found in the Coptic monastery of Phoebammon . Finally, we also have to consider the possibility of the supply of destoned dates and olives. The recovery of secreted plant remains is facilitated by the sampling of concentrations of feces, such as found in cesspits. Judging by the scarcity of water, it is not likely that fl ushed latrines were present in Berenike and Shenshef. During the excavation seasons, relieving oneself is done either in special tents used as lavatories or behind tamarisk hillocks, both situated at isolated spots to diminish the nuisance of fl ies. If this was practiced in a similar way in Roman times, clone rack it implies that only a fraction of fruits such as pomegranate and fig would be retrieved later on by archaeological research from the dump areas in the city proper.The original vegetation in the vicinity of Roman Berenike would not have differed much from the present one.

Climatic change that resulted in modern aridity came about between the First and Fourth Dynasties, in which the period from 2350 to 500 BC, prior to the Persian and Greek rule, was exceptionally dry . In the Eastern Desert, most of the vegetation would have been confined to the wadi branches and the coastal plain. Wadi branches offered occasional runoff water and had a more permanent supply of underground water, and in the coastal plain the vegetation could benefit from seepage and morning dew. Just like today, the sparse herbaceous vegetation had a limited carrying capacity for wild and domestic animals in most years. This picture is supported by the description of the Eastern Desert by Theophrastus and Pliny , who stated that there were no trees except the acacia, which was even sparsely present due to the lack of water. Although the analysis of wood samples from Berenike, Shenshef, and Kalalat does not allow a quantitative interpretation of possible shifts in the arboreal part of the local vegetation, some interesting conclusions can be deduced from the composition of the samples. The charcoal analysis from an early Ptolemaic industrial dump area of brick-making debris revealed that the dominant species was mangrove , in association with a minor proportion of members of the goose foot family . It is most likely, that we are dealing here with Suaeda monoica, the only woody chenopod of a reasonable size that is still present in small populations on the coastal plain. Its massive exploitation for firewood during the earliest habitation period of Berenike can be explained by the clearance of the local area to facilitate the layout of the area meant for habitation and harbor facilities.

This cutting down offered a temporary supply of low-quality firewood, as was evidenced by several fire experiments. This kind of exploitation is supported by the puffing structures in the charcoal of the Goose foot species, which indicates that fresh, living wood was used . Also in Roman contexts, charcoal and even worked wood and wood chips of Avicennia marina are represented in reasonable quantities. A second mangrove species, Rhizophora mucronata Lam., is also evidenced by wood remains , confirming that a mixed mangrove vegetation was present, possibly even at a closer distance to Berenike than that of today. The most dominant tree in Roman contexts that has been used for making charcoal is the acacia. This tree is still highly valued by nomads as its dense wood easily ignites, has a high caloric value, and does not easily fall apart in a charred condition. Judging by the present vegetation, it is most likely that A. tortilis was used as a local source of fuel. Additionally, charcoal from other acacia species might have been imported from the relatively densely wooded Gebel Elba. It remains an open question as to what extent local acacia trees were exploited. If only dead branches were gathered, as is the current practice among the nomads, the exploitation would have had no impact on their proportion in the vegetation. At the same time, such a selective exploitation would not have matched the demand. Harvesting of most of the tree is even possible, as the plant coppices well, so that there is no need for replanting. But if trees are exploited that way, a harvesting schedule based on a 10-year rotation period is necessary . Moreover, a reasonably dense population should have been available to fulfill the demand, a situation that certainly did not exist. Over exploitation of trees would have resulted in their virtual disappearance within a short period of time. In that case, the current trees have to be considered as recolonized specimens, whose spread must have been a gradual process because the mountain ridges are natural barriers for wind dispersal, and the alternative dispersal by camels takes some time.

According to Oedekoven , many wadis in the Eastern Desert and the Sinai still bear names of trees that once grew there but were intensively cut in the distant past when forests in the Nile Valley were cut in favor of agricultural land. The relict character of the sugar date , the twisted acacia , and the leafl ess tamarix in the Eastern Desert is mentioned by Zahran and Willis . The relict nature of these trees is supported by Hobbs , who refers to bedouins who remember former times in which many more trees were present in particular parts of the desert. Large-scale cutting by local tribes is considered the main reason for over exploitation of densely wooded areas, and prolonged drought periods have a negative impact on the tree cover as well.The Diversity of Cultivated Plants. The cultivated plant species that have been found at Berenike and Shenshef can be categorized according to their possible use: cereals, pulses, vegetables, edible fruits, condiments, oil-yielding plants, and plants used for dyeing and tanning . This categorization is based on the proxy value of the recovered plant remains Although many species can be used in medical treatments, this category is left aside as no clear evidence is available as yet for such use. This is not to say, of course, that as the occasion arose, particular plants might have been used in such a way. Five different cereals have been found at Berenike and Shenshef: hulled six-row barley , hard wheat , sorghum , Johnson grass , and rice . Barley, hard wheat, and sorghum can be considered as staple foods. Despite the relatively low numbers of Johnson grass that were recovered from both sites, the correlation between this potential weed and sorghum is weak, so that it is justifiable to consider it as an independent crop. Also the number of rice kernels and rice chaff is limited, but their presence in both early and late habitation layers indicate that this exotic cereal must have been of special importance. The supply of rice might partly have been intended for an Indian community that possibly dwelled at Berenike in the early Roman period, 4×8 tray grow as suggested by substantial quantities of unearthed Indian-made fine wares and a Tamil-Brahmi graffiti . Papyrological evidence, on the other hand indicates that, at least in the early Roman period, rice was also available in the Fayum. Transport of rice from Berenike further northward to Rome seems, therefore, most likely. Both barley and wheat are represented by grains and threshing remains. The threshing remains consist of chaff and sometimes also fragments of the culm. A quantitative comparison between barley and wheat can be done with respect to both the grains and the rachis nodes. The lemma and palea of both cereals are heavily fragmented in most samples, making it almost impossible to use these fragments for a quantitative comparison of barley and wheat. As for rachis fragments as a basis of comparison, it makes sense to quantify the rachis nodes and not the rachis internodes because the spikelets, in which the grains develop, are attached to the rachis nodes. Therefore, the number of rachis nodes is interchangeable with the number of grains. In comparing six-row barley with hard wheat, each rachis node of barley corresponds with three-grain kernels and each rachis node of hard wheat with an average number of three grain kernels that develop within a spikelet. As shown in Table 4.1, the numbers of grains and rachis nodes are not consistent.

The numbers of grains are relatively low with the exception of barley grains from the early habitation period, which even outnumber the rachis nodes. The rachis nodes seem to present a more reliable picture, all the more so because their numbers are much higher. Based on these rachis nodes, the general picture for Berenike is that the proportion of barley to wheat is 0.6 to 0.8. In Shenshef, barley predominates and its proportion to wheat is 1.2 to 1.0. In contrast with barley and wheat, the inflorescence of sorghum is a panicle in which the fertile spikelets are almost sessile. With the exception of a single grain kernel, all the threshing remains of sorghum that were found consisted of lemmas and paleas only. Lemmas and paleas of sorghum are tougher than those of barley and wheat and for this reason are mostly complete specimens. Hence, the quantification of sorghum could be based on the counting of the lemmas and paleas . Sorghum was only available during the second habitation period. The single chaff fragment recovered from an early context is considered as an intrusion. Judging by the absolute numbers of barley and wheat, the highest consumption of these cereals can be ascribed to the first and second centuries AD. In the second habitation period, barley and wheat were partly replaced by sorghum. Although there is some overlap with respect to the dating of fourth to early sixth centuries AD, it seems that the consumption of cereals had diminished in the last part of this habitation period, which is especially true for Berenike. Although cereals might have been locally cultivated on a limited scale, in which sorghum in particular might have produced some decent yields in years with sufficient winter rains, the bulk of these staple grains would have been obtained from the productive Nile Valley. Based on the analysis of the cereals and their accompanying weed flora, it appears that samples from Berenike that resemble those from Shenshef the most originate from trench BE94-1 , trench BE95-10 and trench BE96- 16 . These trenches are located in the dump area in the northwestern part of the site and all samples are dated to the fi fth century and early sixth centuries AD, the period that coincides with the occupation of Shenshef. The group of pulses is represented by the lentil , the white lupin , the faba bean , the bitter vetch , the mung bean , the grass pea , the chickpea , and the Abyssinian pea . Only the lentil, the white lupin, the faba bean, and the bitter vetch are recorded from both Berenike and Shenshef, and their number of plant remains indicates that the lentil and the white lupin were the primary staple. So far, the other four pulses are only found at Berenike and include import from abroad. This is true for the mung bean, which originates from India, and the Abyssinian pea, which originates from Ethiopia. Like the cereals, the pulses that were part of the staple foods also would have been brought in from the Nile Valley. Vegetables are underrepresented at Berenike and Shenshef, as is the case in most archaeobotanical records. This is especially true for green vegetables, which are mostly harvested before seeds are produced, such as lettuce . Seeds from this species have been found only occasionally and might concern seed stock or, more likely, are produced by bolted lettuce. This makes it difficult to decide to what extent such vegetables were available to the inhabitants of Berenike and Shenshef.

Therefore, longer-term research is required to develop non-chemical alternatives, and their adaptation will require effective integration with other methods of disease, pest, and crop management . In the post-methyl bromide era, Verticillium wilt is likely to reemerge as a major disease for conventional strawberry production. The disease already is a major problem in some organic production fields. In strawberry, symptoms begin to appear during early to mid-season, with outer leaves on infected plants turning yellow, drooping, and later turning brown and dry. Yield from these affected plants can be dramatically reduced and infected plants usually die before the end of the season . The fungus survives in the soil as microsclerotia for many years, and survives better in sandy loam soils typical of strawberry production fields in coastal Califtornia than in other types of soil . Large numbers of microsclerotia are formed in colonized Thissue of susceptible crops, and a few are formed even on non-hosts . Whether microsclerotia are formed on infected strawberry plants is not known. Resistance to Verticillium wilt is unavailable in currently used commercial cultivars and tolerance in these cultivars is low. With the phase out of methyl bromide and possible future loss or restrictions on the use of alternative fumigants, resistance to Verticillium wilt has now become a selection criterion in some breeding programs.

As a result, flower harvest buckets resistance to Verticillium wilt in locally adapted strawberry cultivars may increase over time. The concept of rotating crops to manage plant diseases is perhaps one of the oldest cultural practices in agriculture . The utility of this practice in reducing Verticillium dahliae inoculum and subsequent disease intensity has been equivocal . Microsclerotia of V. dahliae survive in the soil up to 10 years, and the extensive host range and lack of host specificity reduce the usefulness of some crop rotations for Verticillium wilt management . However, recent work has shown that rotations with broccoli dramatically reduce microsclerotial numbers and Verticillium wilt incidence in susceptible crops. If rotations of broccoli are successful in strawberry, they will be equally applicable to both conventional and organic strawberry production systems. Although the benefits of rotations are numerous and quantification of these benefits in dollar terms is difficult, simple cost-benefit analysis of adapting rotations will inevitably lead to a better understanding of their composite benefits. Such information also may lead to increased adoption of crop rotations. The objectives of this study were to determine the effect of crop rotation on soil borne fungal inoculum density, disease severity, and strawberry growth and yield; to assess the effectiveness of crop rotation in soil with no detectable Verticillium spp. to improve strawberry growth and yield; and to obtain a cost-benefit analysis of this method of managing Verticillium wilt in strawberry.

Experiments on vegetable–strawberry rotations were conducted at two sites from winter 1997 to fall 2000. One site was located at the United States Department of Agriculture, Agricultural Research Station at Salinas, CA with no history of either strawberry cultivation or Verticillium wilt ; however, the site had other soilborne pathogens such as Pythium spp., Rhizoctonia solani, binucleate Rhizoctonia, and Cylindrocarpon spp. The other site was located at the MonTherey Bay Academy in Watsonville, CA and has a history of strawberry production and disease pressure from V. dahliae as well as the above-noted root rot pathogens. Both sites lie in the major strawberry production areas of the central coast of Califtornia and prior investigations confirmed that black root rot pathogens were present and causing disease . Soil at the Salinas site was a Chualar loam with 57% sand, 31% silt, 1.1% organic matter, and a pH of 5.6. Soil at the Watsonville site was an Elder sandy loam with 62% sand, 26% silt, 0.6% organic matter, and a pH of 6.1. Average yearly precipitation at Salinas and Watsonville was 429 and 757 mm, respectively, and mean daily temperature ranged from 8.3 to 19.9°C and 8.3 to 19.7°C , respectively, during the study. The experimental design was a randomized complete block design with four replications. At both locations, two crops per year of vegetables were grown followed by rotation with annual strawberry. There were three rotation treatments at each location in 1997 and 1999. The crop sequences were broccoli-broccoli-strawberry, lettuce lettuce-strawberry and Brussels sprouts-strawberry or cauliftlower-cauliftlower-strawberry . The first cycle of vegetables, including broccoli , cauliftlower , Brussels sprouts , and lettuce , was grown in January or February and harvested in June in each year. The second cycle of vegetables was planted in July and harvested in September or October.

Because of varying maturity times, planting dates were staggered so that all crops could be harvested and incorporated at the same time. At the Watsonville site, the individual plots consisted of two beds of 7.6 m in length with a single plant line for Brussels sprouts and two seed lines on the 1-m-wide beds standard for lettuce and broccoli production in coastal Califtornia. Plant spacing was approximately 30 cm between plant lines and 20 cm between plants within a plant line for lettuce and Brussels sprouts and 10 cm between plants for broccoli. At the Salinas site, the individual plots were 9 m long and eight 1-m beds wide and consisted of two plant lines for lettuce and broccoli production in coastal Califtornia. The beds contained a single plant line for cauliftlower. Plant spacing was approximately 30 cm between plant lines and 30 cm between plants within a plant line for lettuce and cauliftlower. Plant spacing was 15 cm between plants for broccoli. In both years, 4- to 5-week-old transplants of broccoli, cauliftlower, Brussels sprouts, and lettuce were planted. At the Salinas site, however, lettuce was direct seeded in two rows per bed and thinned to a spacing of 30 cm 3 weeks after emergence. After transplanting, the experimental sites were irrigated with sprinklers for 3 to 4 days to ensure proper establishment of plants. The rotational vegetable planting cycles were timed to include two rotational crops of broccoli, cauliftlower, or lettuce per year and one Brussels sprouts crop. All experiments were maintained using standard cultural practices for coastal Califtornia vegetable production. Preplant fertilizer at 450 kg ha–1 was applied to all beds before transplanting or direct seeding, and all plots were drip-irrigated twice a week. Two side dressings at 67 kg ha–1 followed by fertigation at the rate of 11 to 22 kg ha–1 were provided during each season. At maturity, all marketable vegetables were harvested and remaining residues were flail shredded, air dried on the soil surface for 48 h, and incorporated into the soil to a depth of 15 to 20 cm using a rototiller. Three to four weeks after incorporation, the beds in all plots were reworked for the next vegetable or strawberry production cycle. The second crops of vegetables were grown and incorporated similarly.Standard cultural practices for strawberry production were followed for bed preparation . Beds in the control plots were fumigated with methyl bromide plus chloropicrin at the rate of 450 kg ha–1. Fumigants were injected at a depth of 15 to 25 cm through two hollow shanks, and black or brown high-density polyethylene tarps were used to cover the soil immediately after application. Vegetable rotation plots also were covered with black or brown high-density polyethylene tarps before planting strawberry. Two weeks after fumigation, slow release fertilizer was applied at the rate of 672 kg ha–1 at both sites to all treatments. Fertilizer was band placed about 10 to 15 cm deep in each planting row and covered with approximately 4 cm soil to prevent direct contact with plant roots. Bare-root transplants of strawberry cv. Selva were planted 15 cm apart in all plots at both locations in two rows per bed spaced 40 cm apart in mid November of 1997–98 and 1999–2000 . The drip irrigation system consisted of two drip tapes with emitters spaced 30 cm apart, placed 8 cm from the bed center at a depth of 2 to 4 cm, with an emitter flow rate of 0.87 liter min–1 at 70 kPa. Approximately 30 mm of water per week was applied to each of the experimental plots. After plants were established, round flower buckets drip irrigation was applied once or twice per week for 1.5 to 2 h depending on soil moisture conditions. Hand weeding was done periodically when necessary. All management and harvest of the strawberry crop was done by cooperating commercial growers. To determine the densities of propagules of V. dahliae and Pythium spp., soil samples were collected at the beginning and end of the rotation crop and every month after the start of strawberry production.

Soil samples were taken to a depth of 6 to 10 cm from 10 random sites in each treatment, in the center four beds at the Salinas site and in both beds at the Watsonville site. Samples from each treatment were bulked and placed in paper bags and air dried for 4 weeks on greenhouse benches. The dried soil then was assayed for V. dahliae propagules by direct plating onto improved NP-10 medium using the modified Anderson sampler technique . After 3 weeks of incubation in the dark, the plates were washed in a gentle stream of water and examined under a sThereoscope . The number of microsclerotial colonies on each plate was counted and expressed as microsclerotia per gram of dry soil. For the determination of the number of CFU of Pythium spp., 1 g of air-dried soil was added to 10-ml sterilized water blanks and vortexed to mix the soil, and 500 µl of this soil solution was distributed evenly over a petri dish containing a semiselective Pythium medium . The medium consisted of corn meal agar amended immediately after autoclaving with 0.1% Tween 20 followed by pimaricin , ampicillin , rifampicin , rose bengal , and Benomyl 50WP after the medium had cooled to 50°C . After 24 h of incubation at 25°C, the surfaces of the plates were washed free of soil under a gentle stream of water and the number of colonies counted. Plates were placed back into the incubator and subsequent counts made again after 24 h. The total colony counts were expressed as CFU g–1 of dry soil and the presented data reflects the mean of the three replicates. Prior investigations at these sites revealed that Pythium ultimum was the most commonly encounThered species, but other species such as P. irregulare were recovered as well .To determine the relative effects of different rotation treatments, strawberry plant growth was monitored by recording the plant canopy diameter of 20 arbitrarily chosen plants per replication at least twice per season in each replication. Measurements were made in both the east-west and north-south directions because there were significant directional effects early in the season. Development of Verticillium wilt at Watsonville was assessed based on foliar symptoms . The symptoms of root diseases such as black root rot complex were similar but rarely progressed past stunting with leaf chlorosis later in the season . Assessments were made at 2-week intervals starting from June until the end of the season . No Verticillium wilt was observed at the Salinas site. Twenty plants per plot were visually rated to monitor disease progress. The disease severity estimate was made based on a scale of 1 to 8, where 1 = healthy plant, 2 = moderately stunted, 3 = moderately stunted and slight outer rosette of dead leaves, 4 = moderately stunted and moderate outer rosette of dead leaves, 5 = significantly stunted and slight outer rosette of dead leaves, 6 = significantly stunted and moderate outer rosette of dead leaves, 7 = significantly stunted and significant rosette of dead leaves, 8 = dead plant. Data on the cost of production of strawberry fruit, including fumigating, preparing of land, mulching, planting, irrigating, fertigating, weeding, and fruit harvesting, were obtained from four growers that produce both strawberry and vegetables. Similarly, total yields per unit area of production, corresponding revenue earned, and so on also were obtained. Similar data were obtained for strawberry production without fumigation. The cost of producing two crops of broccoli, revenue from broccoli, and the following strawberry crop also was obtained.