Characterization and quantification of anthocyanins and polyphenolics in bluehHoneysuckle (Lonicera caerulea L.)

Anthocyanins and phenolics of 10 blue honeysuckle (Lonicera caerulea L.) genotypes were characterized and quantified by HPLC-DAD. Peak assignments were confirmed by low-resolution electrospray mass spectrometry. Six anthocyanins were detected with the major peak identified as cyanidin 3-glucoside. Five additional anthocyanins were characterized as cyanidin 3,5-diglucoside, cyanidin 3-rutinoside, pelargonidin 3-glucoside, peonidin 3-glucoside, and peonidin 3-rutinoside. Four polyphenolics were identified as chlorogenic acid, neochlorogenic acid, quercetin 3-rutinoside, and quercetin 3-glucoside. Two additional unidentified phenolics were characterized as flavonol and hydroxycinnamic derivatives based on UV-vis spectra. Hydroxycinnamate levels ranged from 30.4 to 156.2 mg/100 g, whereas the flavonol content ranged from 12.6 to 32.8 mg/100 g. The L. caerulea subspecies boczkarnikovae contained the highest amounts of hydroxycinnamic derivatives and flavonols.     

Performance of a short tool to assess dietary intakes of fruits and vegetables, percentage energy from fat and fibre

OBJECTIVES: We describe the methods used to develop and score a 17-item 'screener' designed to estimate intake of fruit and vegetables, percentage energy from fat and fibre. The ability of this screener and a food-frequency questionnaire (FFQ) to measure these exposures is evaluated. DESIGN: Using US national food consumption data, stepwise multiple regression was used to identify the foods to be included on the instrument; multiple regression analysis was used to develop scoring algorithms. The performance of the screener was evaluated in three different studies. Estimates of intakes measured by the screener and the FFQ were compared with true usual intake based on a measurement error model. SETTING: US adult population. SUBJECTS: For development of instrument, n=9323 adults. For testing of instrument, adult men and women in three studies completing multiple 24-hour dietary recalls, FFQ and screeners, n=484, 462 and 416, respectively. RESULTS: Median recalled intakes for examined exposures were generally estimated closely by the screener. In the various validation studies, the correlations between screener estimates and estimated true intake were 0.5-0.8. In general, the performances of the screener and the full FFQ were similar; estimates of attenuation were lower for screeners than for full FFQs. CONCLUSIONS: When coupled with appropriate reference data, the screener approach described may yield useful estimates of intake, for both surveillance and epidemiological purposes.     

In vivo investigation of changes in biomarkers of oxidative stress induced by plant food rich diets

It is well established that vegetables and fruit (VF) contain antioxidant phytochemicals. Consequently, it is expected that individuals who consume diets with a high content of VF should be better protected against oxidative cellular damage than individuals who do not, but not all data support this assumption. The objective of this study was to identify possible explanations for this conundrum. The effects of two diets that differed in VF content on markers of oxidative damage were studied. Sixty-four women participated in a 14-day dietary intervention. Subjects consumed on average either 3.6 or 12.1 servings of VF per day. The primary end points assessed were 8-hydroxy-2-deoxyguanosine (8-oxo-dG) in peripheral lymphocyte DNA and 8-isoprostaglandin F-2alpha (8-iso-PGF2alpha) excreted in urine. Subjects consuming the high versus low VF diet had lower concentrations of 8-oxo-dG (p < 0.01) and of 8-iso-PGF2alpha (p < 0.01). However, the reduction in oxidative end points by high VF was not uniform. Rather, an antioxidant effect was observed primarily in individuals whose oxidative end points at baseline were above the median for the study population. Using change in plasma carotenoids (end point minus baseline measurements) as an index of phytochemical intake, the reduction in oxidative markers was inversely proportional to change in plasma carotenoids; this effect was stronger for lipid peroxidation (p < 0.01) than DNA oxidation (p < 0.05). These findings imply that increasing exogenous antioxidant exposure may primarily benefit individuals with elevated levels of oxidative stress. Null findings do not necessarily indicate that an antioxidant compound lacks in vivo activity.     

Residue depletion study and withdrawal period for flunixin-N-methyl glucamine in bovine milk following intravenous administration

The objective of this study was to establish a withdrawal period for flunixin in milk by quantifying 5-hydroxyflunixin, the marker residue, in bovine milk as a function of time, following intravenous treatment of lactating dairy cows with flunixin-N-methyl glucamine (Banamine or Finadyne). Lactating dairy cows were dosed on three consecutive days at 2.2 mg of flunixin free acid/kg of body weight/day. Milk was collected twice daily and assayed using a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) procedure. The method was validated at concentrations in the range 0.5-250 ppb. The concentrations for 5-hydroxyflunixin measured 12 h after the last administration of drug ranged from 1.56 to 40.6 ppb for all cows. Milk concentrations for 5-hydroxyflunixin were used to establish withdrawal periods of 36 h using guidelines established by the U.S. Food and Drug Administration/Center for Veterinary Medicine and 24 h using guidelines established by the European Medicinal Evaluation Agency/Committee on Veterinary Medicinal Products.     

Optimization of chick embryotoxicity bioassay for testing toxicity potential of fungal metabolites

The optimization of a simple, sensitive procedure using a chick embryotoxicity screening test (CHEST) bioassay for detection of toxic compounds is presented. Dosing protocols of eggs, using several mycotoxins (aflatoxin B1, deoxynivalenol, T-2 toxin) and appropriate controls, were evaluated for embryonic sensitivity, overall practicality of the procedure, and consistency of results. It was found that both type of carrier solvent and volume injected could significantly affect overall embryonic mortality. The chick embryo was most sensitive to the effects of toxins and solvents after 1 or 2 days of incubation; a rapid decrease in response was observed as the age of the embryo at dosing increased. Following administration of the toxins just below the shell membrane by way of a small hole (less than 0.5 mm diameter) punched in the shell, a good dose-response (% mortality) could be obtained regardless of the site of injection (except directly into the yolk), although dosing via the air sac position resulted in a slightly better statistical outcome. Although some variations in calculated LD50 values were found among repeated assays, statistical analyses showed that the differences were not due to dosing protocol but to the variations in embryo sensitivities among batches of eggs. Thus, if standard reference toxins for comparison are run concurrently, the CHEST assay can prove to be a very satisfactory model, as well as having considerable flexibility to be adapted to the needs and resources of many laboratories.     

Liquid chromatographic method for quantitation of glyphosate and metabolite residues in organic and mineral soils, stream sediments, and hardwood foliage

A liquid chromatographic method for determining glyphosate (GLYPH) and its major metabolite aminomethylphosphonic acid (AMPA) in various environmental substrates is described. Ion-exchange column chromatography is coupled with post-column ninhydrin derivatization and absorbance detection at 570 nm. Use of a valve-switching technique allowed quantitation of both analytes in a single chromatographic run and eliminated slow-eluting, coextracted interferences. The method was successfully used to quantitate GLYPH and AMPA in organic and mineral soils, stream sediments, and foliage of 2 hardwood brush species. Mean recovery efficiencies for GLYPH as determined from fortified blank field samples were as follows: bottom sediment 84%, suspended sediment 66%, organic soils 79%, mineral soils 73%, alder leaf litter 81%, salmonberry leaf litter 84%, and artificial deposit collectors 87%. Precision for GLYPH determination was good with less than 14% coefficient of variation on mean recovery for all substrates. Limits of detection were lowest for sediments (0.01 microgram/g dry mass) and highest for foliage substrates (0.10 microgram/g dry mass). Using this system, 6 samples/person/day were routinely analyzed.     

Letters

Editor's note: The AVJ welcomes letters from members in all areas of the profession on matters of importance to you. Please keep them brief ‐ to meet our space constraints. Letters will be subject to minimal editing procedures. Subject to letters complying with the AVJ's legal responsibilities, they will not be censored. Nor will individuals or groups waging'‘campaigns’ be permitted to abuse these pages. If submitting a letter intended for publication, kindly identify it as such. Letters to the Editor can be sent by mail, fax or e‐mail at the contact points listed at the start of the News Section. Writers may use a pseudonym to protect their identities ‐ but must supply the Editor with verifiable names and points of contact.     

Movement rules for herbivores in spatially heterogeneous environments: responses to small scale pattern

Foraging herbivores respond to the spatial pattern of resources at a variety of scales. At small scales of space and time, existing models capture the essence of the feeding process and successfully predict intake rates. Models that operate over larger scales have not exhibited a similar success, in part because we have a limited understanding of the rules used by animals to make decisions in spatially complex environments, or of the consequences of departing from these rules. To evaluate the rules that large herbivores use when navigating between forages, we examined movements of bighorn sheep foraging on apparent prey (alfalfa plants) in hand-constructed patches of plants. Observations of movements and path lengths were compared to simulations that used a variety of different rules-of-thumb to determine a search path. Rules used in simulations ranged from a random walk with various detection distances, to more complicated rules that solved a variant of the travelling salesman problem. Simulations of a random walk yielded movement lengths that exceeded observations by a factor of 3 for long detection distances, and by 30-fold for short detection distances. Observed move distances were most closely approximated by simulations based on a nearest-neighbor ruleover 75 % of all moves by bighorn sheep were to the closest available plant. Movement rules based on random walks are clearly inappropriate for many herbivores that typically consume visually apparent plants, and we suggest the use of a nearest-neighbor rule for modelling foraging by large herbivores.     

Hepatic serine and alanine metabolism during endotoxin-induced fever in sheep.

Time course changes in plasma amino acid concentrations and the hepatic metabolism of serine and alanine were measured in six mature wethers during endotoxin-induced fever. In separate trials, the animals' responses to injections of saline and endotoxin were measured. The endotoxin was from Escherichia coli serotype 055:B5 and was injected intravenously (4 micrograms/kg body weight). Liver biopsies were obtained from the sheep at 6 h postinjection during both endotoxin and saline injection trials. Rectal temperature in the endotoxin treated animals was increased (P less than 0.05, above that in control animals from 4.25 h to 9 h postinjection, with a maximum rise of 2.43 degrees C at 5.5 h postinjection. Glucose concentration in jugular plasma decreased (P less than 0.05) by 3 h postinjection and remained depressed throughout the 24 h postinjection sampling period. Plasma serine concentration was decreased (P less than 0.05) by 3 h postinjection. Plasma alanine concentration was decreased significantly (P less than 0.05) only at 24 h postinjection. Endotoxin injection increased (P less than 0.05) hepatic oxidation of 14C-serine (162%) and the net incorporation of 14C-serine carbon into hepatic protein (173%) and glycogen (275%). The net incorporation of 14C-alanine carbon into hepatic protein (172%) and glycogen (323%) were increased (P less than 0.05) by endotoxin injection, while alanine oxidation was not affected by endotoxin treatment (P greater than 0.05). The increased hepatic use of serine may explain, in part, the dramatic decrease in plasma concentrations of this amino acid following endotoxin injection into sheep.