Mark R. Ellersieck

Restoring a maize root signal that attracts insect-killing nematodes to control a major pest

When attacked by herbivorous insects, plants emit volatile compounds that attract natural enemies of the insects. It has been proposed that these volatile signals can be manipulated to improve crop protection. Here, we demonstrate the full potential of this strategy by restoring the emission of a specific belowground signal emitted by insect-damaged maize roots. The western corn rootworm induces the roots of many maize varieties to emit (E)-β-caryophyllene, which attracts entomopathogenic nematodes that infect and kill the voracious root pest. However, most North American maize varieties have lost the ability to emit (E)-β-caryophyllene and may therefore receive little protection from the nematodes. To restore the signal, a nonemitting maize line was transformed with a (E)-β-caryophyllene synthase gene from oregano, resulting in constitutive emissions of this sesquiterpene. In rootworm-infested field plots in which nematodes were released, the (E)-β-caryophyllene-emitting plants suffered significantly less root damage and had 60% fewer adult beetles emerge than untransformed, nonemitting lines. This demonstration that plant volatile emissions can be manipulated to enhance the effectiveness of biological control agents opens the way for novel and ecologically sound strategies to fight a variety of insect pests.

Identification of Fecal Escherichia coli from Humans and Animals by Ribotyping

ABSTRACT Fecal pollution of water resources is an environmental problem of increasing importance. Identification of individual host sources of fecal Escherichia coli, such as humans, pets, production animals, and wild animals, is prerequisite to formulation of remediation plans. Ribotyping has been used to distinguish fecalE. coli of human origin from pooled fecal E. coli isolates of nonhuman origin. We have extended application of this technique to distinguishing fecal E. coli ribotype patterns from human and seven individual nonhuman hosts. Classification accuracy was best when the analysis was limited to three host sources. Application of this technique to identification of host sources of fecal coliforms in water could assist in formulation of pollution reduction plans.

Increased survival of western corn rootworm on transgenic corn within three generations of on-plant greenhouse selection.

To delay evolution of insect resistance to transgenic crops producing Bacillus thuringiensis (Bt) toxins, nearby “refuges” of host plants not producing Bt toxins are required in many regions. Such refuges are expected to be most effective in slowing resistance when the toxin concentration in Bt crops is high enough to kill all or nearly all insects heterozygous for resistance. However, Bt corn, Zea mays, introduced recently does not meet this “high-dose” criterion for control of western corn rootworm (WCR), Diabrotica virgifera virgifera. A greenhouse method of rearing WCR on transgenic corn expressing the Cry3Bb1 protein was used in which approximately 25% of previously unexposed larvae survived relative to isoline survival (compared to 1–4% in the field). After three generations of full larval rearing on Bt corn (Constant-exposure colony), WCR larval survival was equivalent on Bt corn and isoline corn in greenhouse trials, and the LC50 was 22-fold greater for the Constant-exposure colony than for the Control colony in diet bioassays with Cry3Bb1 protein on artificial diet. After six generations of greenhouse selection, the ratio of larval recovery on Bt corn to isoline corn in the field was 11.7-fold greater for the Constant-exposure colony than the Control colony. Removal from selection for six generations did not decrease survival on Bt corn in the greenhouse. The results suggest that rapid response to selection is possible in the absence of mating with unexposed beetles, emphasizing the importance of effective refuges for resistance management.

Comparison of Ribotyping and Repetitive Extragenic Palindromic-PCR for Identification of Fecal Escherichia coli from Humans and Animals

ABSTRACT This report compares the performances of two popular genotypic methods used for tracking the sources of fecal pollution in water, ribotyping and repetitive extragenic palindromic-PCR (rep-PCR). The rep-PCR was more accurate, reproducible, and efficient in associating DNA fingerprints of fecal Escherichia coli with human and animal hosts of origin.

Effects of temperature on the performance of finishing swine: II. Effects of a cold, diurnal temperature on average daily gain, feed intake, and feed efficiency

Ninety-six crossbred barrows and gilts weighing 90 +/- .67 kg were used during a 21-d study to determine the effects of a hot, diurnal temperature (H; 22.5 to 35 degrees C) compared with a constant, thermoneutral temperature (TN; 20 degrees C) and the effects of sex (barrows vs gilts) on performance. A secondary objective included the determination of weight loss as a result of a 24-h fast immediately after the 21-d feeding study of commingled vs not commingled hogs of both environmental treatments (TN and H). Pigs housed in the hot, diurnal temperature gained 16.3% more slowly (P less than .001;.77 vs. .92 kg/d) than those in the constant, thermoneutral environment. Feed intake (FI) for the H pigs was 10.9% less (P less than .001; 3.01 vs 3.38 kg/d) than that for the TN pigs. The H pigs gained 17.6 g/d less and consumed 43.5 g/d less feed for every C degrees above 20 degrees C; however, no differences were observed for feed efficiency (F/G; 3.86 vs 4.19 kg for the TN and H pigs, respectively). Average daily gain and feed/gain (F/G) were not affected by sex. Likewise, no significant interactions of temperature x sex were observed for ADG, FI, or F/G. Weight loss (shrinkage) during the 24-h fast was not affected by commingling; however, the H pigs lost 17.5% more weight (P less than .05) than the TN pigs (3.82 vs 3.25%, respectively).

Sex and dose-dependent effects of developmental exposure to bisphenol A on anxiety and spatial learning in deer mice (Peromyscus maniculatus bairdii) offspring.

Bisphenol A (BPA) is a widely produced, endocrine disrupting compound that is pervasive in the environment. Data suggest that developmental exposure to BPA during sexual differentiation of the brain leads to later behavioral consequences in offspring. Outbred deer mice (Peromyscus maniculatus bairdii) are an excellent animal model for such studies as they exhibit well-defined sex- and steroid-dependent behaviors. Here, dams during gestation and lactation were fed with a phytoestrogen-free control diet, the same diet supplemented with either ethinyl estradiol (0.1 ppb), or one of the three doses of BPA (50 mg, 5 mg, 50 μg/kg feed weight). After weaning, the pups were maintained on control diet until they reached sexual maturity and then assessed for both spatial learning capabilities and anxiety-like and exploratory behaviors. Relative to controls, males exposed to the two upper but not the lowest dose of BPA demonstrated similar impairments in spatial learning, increased anxiety and reduced exploratory behaviors as ethinyl estradiol-exposed males, while females exposed to ethinyl estradiol, but not to BPA, consistently exhibited masculinized spatial abilities. We also determined whether dams maintained chronically on the upper dose of BPA contained environmentally relevant concentrations of BPA in their blood. While serum concentrations of unconjugated BPA in controls were below the minimum level of detection, those from dams on the BPA diet were comparable (5.48±2.07 ng/ml) to concentrations that have been observed in humans. Together, these studies demonstrate that developmental exposure to environmentally relevant concentrations of BPA can disrupt adult behaviors in a dose- and sex-dependent manner.

Holding Thermal Receipt Paper and Eating Food after Using Hand Sanitizer Results in High Serum Bioactive and Urine Total Levels of Bisphenol A (BPA)

Bisphenol A (BPA) is an endocrine disrupting environmental contaminant used in a wide variety of products, and BPA metabolites are found in almost everyone’s urine, suggesting widespread exposure from multiple sources. Regulatory agencies estimate that virtually all BPA exposure is from food and beverage packaging. However, free BPA is applied to the outer layer of thermal receipt paper present in very high (∼20 mg BPA/g paper) quantities as a print developer. Not taken into account when considering thermal paper as a source of BPA exposure is that some commonly used hand sanitizers, as well as other skin care products, contain mixtures of dermal penetration enhancing chemicals that can increase by up to 100 fold the dermal absorption of lipophilic compounds such as BPA. We found that when men and women held thermal receipt paper immediately after using a hand sanitizer with penetration enhancing chemicals, significant free BPA was transferred to their hands and then to French fries that were eaten, and the combination of dermal and oral BPA absorption led to a rapid and dramatic average maximum increase (Cmax) in unconjugated (bioactive) BPA of ∼7 ng/mL in serum and ∼20 µg total BPA/g creatinine in urine within 90 min. The default method used by regulatory agencies to test for hazards posed by chemicals is intra-gastric gavage. For BPA this approach results in less than 1% of the administered dose being bioavailable in blood. It also ignores dermal absorption as well as sublingual absorption in the mouth that both bypass first-pass liver metabolism. The elevated levels of BPA that we observed due to holding thermal paper after using a product containing dermal penetration enhancing chemicals have been related to an increased risk for a wide range of developmental abnormalities as well as diseases in adults.

Comparison of Serum Bisphenol A Concentrations in Mice Exposed to Bisphenol A through the Diet versus Oral Bolus Exposure

Background: Bisphenol A (BPA) is a widely produced endocrine-disrupting chemical. Diet is a primary route of exposure, but internal exposure (serum concentrations) in animals and humans has been measured only after single oral bolus administration. Objective: We compared serum concentrations of BPA over a 24-hr period after oral bolus administration or ad libitum feeding in mice and assessed for buildup with dietary exposure. Methods: Adult female mice were administered [dimethyl-d6]-BPA (BPA-d6) as a single oral bolus (20 mg/kg body weight) or fed a diet containing 100 mg BPA-d6/kg feed weight ad libitum for 1 week. Serum concentrations were analyzed using isotope dilution liquid chromatography coupled with electrospray tandem mass spectrometry and compared between exposure groups over the first 23 hr and after 7 days of dietary exposure. Results: Maximum concentration (Cmax) for BPA-d6 during the first 24 hr was reached at 1 hr and 6 hr for oral bolus and diet groups, respectively. Relative BPA-d6 bioavailability (unconjugated BPA-d6) was higher in diet-exposed mice than in the bolus group despite a relative lower absorption, a phenomenon consistent with an inhibitory effect of food on first-pass hepatic metabolism. In mice with ongoing dietary exposure, unconjugated BPA-d6 was higher on day 7 than on day 1. Conclusions: This is the first report of serum BPA concentrations in an animal model exposed to this chemical via the diet. Although bolus administration of BPA-d6 led to peak concentrations within 1 hr, Cmax for diet-exposed mice was delayed for several hours. However, bolus administration underestimates bioavailable serum BPA concentrations in animals—and presumably humans—than would result from dietary exposure. Exposure via diet is a more natural continuous exposure route than oral bolus exposure and is thus a better predictor of BPA concentrations in chronically exposed animals and humans.

Role of Egg Density on Establishment and Plant-to-Plant Movement by Western Corn Rootworm Larvae (Coleoptera: Chrysomelidae)

Abstract The effect of egg density on establishment and dispersal of larvae of the western corn rootworm, Diabrotica virgifera virgifera LeConte, was evaluated in a 3-yr field study. Implications of these data for resistance management plans for Bt crops are discussed. Viable egg levels of 100, 200, 400, 800, and 1,600 eggs per infested plant were evaluated in 2000, 2001, and 2002. A 3,200 viable egg level was also tested in 2001 and 2002. All eggs were infested on one plant per subplot in a field that was planted to soybean, Glycine max (L.), in the previous year. For each subplot, the infested plant, three plants down the row, the closest plant in the adjacent row of the plot, and a control plant at least 1.5 m from any infested plant (six plants total) were sampled. In 2000, there were five sample dates between egg hatch and pupation, and in 2001 and 2002, there were six sample dates. On each sample date, four replications of each egg density were sampled for both larval recovery and plant damage. Initial establishment on a corn plant seemed to not be density-dependent because a similar percentage of larvae was recovered from all infestation rates. Plant damage and, secondarily, subsequent postestablishment larval movement were density-dependent. Very little damage and postestablishment movement occurred at lower infestation levels, but significant damage and movement occurred at higher infestation rates. Movement generally occurred at a similar time as significant plant damage and not at initial establishment, so timing of movement seemed to be motivated by available food resources rather than crowding. At the highest infestation level in 2001, significant movement three plants down the row and across the 0.76 m row was detected, perhaps impacting refuge strategies for transgenic corn.

Stylet Penetration by Adult Homalodisca coagulata on Grape: Electrical Penetration Graph Waveform Characterization, Tissue Correlation, and Possible Implications for Transmission of Xylella fastidiosa

Abstract The sharpshooter Homalodisca coagulata (Say) (Hemiptera: Cicadellidae: Cicadellinae) is an exotic vector of the Pierce’s disease (PD) bacterium, Xylella fastidiosa (Wells et al.), that was first observed in California in 1989. H. coagulata has since greatly increased the threat of PD to the grape industry as well as stone fruit, nursery, and ornamental industries in California. This is the first in a series of articles that together describe how sharpshooter stylet penetration behaviors (especially intricate stylet activities, salivation, and ingestion) control transmission (i.e., acquisition and inoculation) of X. fastidiosa. Herein, we categorized and characterized alternating current electrical penetration graph (EPG) waveforms from glass-winged sharpshooter stylet penetration on petiole of susceptible grape (‘Cabernet Sauvignon’), paying special attention to waveform fine structures that are likely to be the key to detecting the instant of inoculation. We also correlated waveforms with salivary sheath termini in grape tissues. For the first time in any EPG study of leafhopper or planthopper feeding, we demonstrate through case studies of individual probes how to follow the process of stylet penetration step by step as it is occurring, including salivary sheath branching and when the stylets first puncture a xylem cell. Finally, we discuss the implications of our findings for understanding the transmission mechanism of X. fastidiosa, in comparison with hypothesized mechanisms in the literature.