Bemisia tabaci （Hemiptera： Aleyrodidae） nymphal feeding in cotton （Gossypium hirsutum） leaves

We used brightfield electron microscopy （BEM）, differential interference contrast microscopy （DICM）, scanning electron microscopy （SEM）, transmission electron microscopy （TEM）, and confocal laser scanning microscopy （CLSM） to investigate the stylet pathways of Bemisia tabaci during nymphal feeding behavior in cotton leaves beginning with penetration of the abaxial leaf surface and ending with stylets in sieve tubes in phloem tissues. Most nymphal stylets within salivary sheaths penetrating leaf tissues made complex turns and developed more than one salivary sheath branch before ending in sieve tubes. The external morphology of the salivary sheaths and their routes between and through leaf cells are described during the present study. Results showed the presence of the stylet within the sieve tubes. B. tabaci nymphs may remove stylets and feed in different sieve tubes. Ten short movies showing the progression of the stylet penetrations from adaxial surface to the sieve tubes are attached to Figures 8-15. The report and movies can be viewed from the internet. Download the movies to a local drive in your computer first for fast upload. The movies are posted on the website http：//www.ars.usda.gov/Services/docs.htm？docid=14629. The movies can be used as a teaching aid in biology classes.

Fate and transport of theβ-adrenergic agonist ractopamine hydrochloride in soil–water systems

The feed additive ractopamine hydrochloride was fortified at four concentrations into batch vials containing soils that differed in both biological activity and organic matter（OM）.Sampling of the liquid layer for 14 days demonstrated that ractopamine rapidly dissipated from the liquid layer. Less than 20% of the fortified dose remained in the liquid layer after 4 hr,and recoveries of dosed ractopamine ranged from 8 to 18% in the liquid layer at 336 hr. Sorption to soil was the major fate for ractopamine in soil：water systems, i.e., 42%–51% of the dose at14 days. The major portion of the sorbed fraction was comprised of non-extractables; a smaller fraction of the sorbed dose was extracted into water and acetone, portions which would be potentially mobile in the environment. Partitioning coefficients for all soils suggested strong sorption of ractopamine to soil which is governed by hydrophobic interactions and cation exchange complexes within the soil OM. Ractopamine degradation was observed, but to mostly non-polar compounds which had a higher potential than ractopamine to sorb to soil. The formation of volatiles was also suggested. Therefore, despite rapid and extensive soil sorption,these studies indicated a portion of ractopamine, present in manures used to fertilize soils,may be mobile in the environment via water-borne events.

Ractopamine up take by alfalfa (Medicago sativa) and wheat (Triticum aestivum) from soil

Ractopamine is a beta adrenergic agonist used as a growth promoter in swine, cattle and turkeys. To test whether ractopamine has the potential to accumulate in plants grown in contaminated soil, a greenhouse study was conducted with alfalfa（Medicago sativa） and wheat（Triticum aestivum） grown in two soils having different concentrations of organic matter（1.3% and 2.1%）, amended with 0, 0.5, and 10 μg/g of ractopamine. Plant growth ranged from 2.7 to 8.8 g dry weight（dw） for alfalfa, and 8.7 to 40 g dw for wheat and was generally greater in the higher organic matter content soil. The uptake of ractopamine in plant tissues ranged from non-detectable to 897 ng/g and was strongly dependent on soil ractopamine concentration across soil and plant tissue. When adjusted to the total fortified quantities, the amount of ractopamine taken up by the plant tissue was low, 〈 0.01% for either soil.

Development of indirect competitive fluorescence immunoassay for 2,2′,4,4′-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels

An indirect competitive fluorescence immunoassay using a DNA/dye conjugate as antibody multiple labels was developed on 96-well plates for the identification and quantification of 2,2＇,4,4＇-tetrabromodiphenyl ether （BDE-47） in aqueous samples. A hapten, 2,4,2＇- tribromodiphenyl ether-4＇-aldehyde, was synthesized, and was conjugated to bovine serum albumin to form a coating antigen. Specific recognition of the antigen by anti-PBDE antiserum was confirmed by a surface plasmon resonance measurement. In the immunoassay, the coating antigen was adsorbed on a 96-well plate first, and a sample, antiserum and biotinylated goat anti-rabbit secondary antibody were then added and reacted sequentially. A biotinylated, double-stranded DNA with 219 base pairs was attached to the secondary antibody by using streptavidin as a molecular bridge. In situ multiple labeling of the antibody was accomplished after addition of a DNA-binding fluorescent dye, SYBR Green I. The working range of the immunoassay for the BDE-47 standard was 3.1-390 ~tg/L, with an IC50 value of 15.6 Ixg/L. The calculated LOD of the immunoassay is 0.73 Ixg/L. The immunoassay demonstrated relatively high selectivity for BDE-47, showing very low cross-reactivity （〈 3%） with BDE-15, BDE-153 and BDE-209. With a spiked river water sample containing 50 Izg/L BDE-47, quantification by the immunoassay was 41.9 ~tg/L, which compared well with the standard GC-ECD method （45.7 Ixg/L）. The developed immunoassay provides a rapid screening tool for polybrominated diphenyl ethers in environmental samples.