Determination of Dicyclohexylamine in Beeswax by Aqueous Normal Phase Liquid Chromatography Coupled With Tandem Mass Spectrometry.
Dicyclohexylamine (DCH) is an excipient present in commercial formulations of fumagillin (Fumagilin-B® and Fumidil-B®), an antibiotic which is employed in apiculture for the control of nosema disease. DCH has been demonstrated to be stable in harvested honey; however, its fate in the beehive has not been investigated. In this study, DCH residues were determined in beeswax samples collected from brood chambers and honey supers from various apiaries throughout Alberta. The determination of DCH was performed using a simple extraction procedure followed by low-temperature cleanup and finally analysis with aqueous normal phase liquid chromatography-electrospray ionization tandem mass spectrometry. The method was found to yield high accuracy and precision (94.9 ± 1.7% to 110.8 ± 6.6%) for replicate beeswax samples spiked with 4, 80, 360 and 2,000 μg kg-1 of DCH. With a limit of quantification of 1 μg kg-1, 100% of the 61 beeswax samples analyzed were found to contain residues of DCH ranging from 15 to 6,410 μg kg-1. The results indicate that DCH can accumulate in beeswax when Fumagilin-B® or Fumidil-B® is applied in apiculture and that these concentrations can vary significantly among beekeeping operations.
Evaluation of Fumagilin-B® and other potential alternative chemotherapies against Nosema ceranae-infected honeybees (Apis mellifera) in cage trial assays
Fumagilin-B® is the only currently registered chemical treatment available to combat nosema disease in apiculture. Fumagillol, the basic hydrolysis product of fumagillin, two semisynthetic fumagillin analogues, and four in-house purely synthetic compounds which were designed to mimic the mode of action of fumagillin against the methionine aminopeptidase type 2 (MetAP-2) enzyme, was observed to exhibit statistically significant biological activity against Nosema ceranae-infected caged bees. None of these compounds were, however, as effective as Fumagilin-B®. The commercially available thymol and enilconazole also exhibited activity against N. ceranae, with thymol being the most promising chemical treatment other than Fumagilin-B®. High cumulative bee mortality was associated with the therapeutic dosage of Fumagilin-B® during our study, suggesting the need for continued investigation.
THE IRON-ACETOCARMINE METHOD OF FIXING AND STAINING CHROMOSOMES
A short description of this procedure was given in Vol. 55 of the American Naturalist, pp. 573â€”574. The method has now been used almost daily for five years, and certain improvements have suggested themselves. The solution can be prepared by mixing about equal volumes of glacial acetic acid and water. To this is added powdered carmine in excess. The liquid is then â€ ? just brought to boil, cooled, and decanted. To the red solution there are then added a few drops of a solution of ferric hydrate in 50 per cent. acetic acid, to act as a mordant. The amount of iron varies with different objects. Too much produces a precipitate in a short time. The more