Holistic screening of collapsing honey bee colonies in Spain: a case study.

BACKGROUND: Here we present a holistic screening of collapsing colonies from three professional apiaries in Spain. Colonies with typical honey bee depopulation symptoms were selected for multiple possible factors to reveal the causes of collapse. RESULTS: Omnipresent were Nosema ceranae and Lake Sinai Virus. Moderate prevalences were found for Black Queen Cell Virus and trypanosomatids, whereas Deformed Wing Virus, Aphid Lethal Paralysis Virus strain Brookings and neogregarines were rarely detected. Other viruses, Nosema apis, Acarapis woodi and Varroa destructor were not detected. Palinologic study of pollen demonstrated that all colonies were foraging on wild vegetation. Consequently, the pesticide residue analysis was negative for neonicotinoids. The genetic analysis of trypanosomatids GAPDH gene, showed that there is a large genetic distance between Crithidia mellificae ATCC30254, an authenticated cell strain since 1974, and the rest of the presumed C. mellificae sequences obtained in our study or published. This means that the latter group corresponds to a highly differentiated taxon that should be renamed accordingly. CONCLUSION: The results of this study demonstrate that the drivers of colony collapse may differ between geographic regions with different environmental conditions, or with different beekeeping and agricultural practices. The role of other pathogens in colony collapse has to bee studied in future, especially trypanosomatids and neogregarines. Beside their pathological effect on honey bees, classification and taxonomy of these protozoan parasites should also be clarified.

Honeybee colony collapse due to Nosema ceranae in professional apiaries.

Honeybee colony collapse is a sanitary and ecological worldwide problem. The features of this syndrome are an unexplained disappearance of adult bees, a lack of brood attention, reduced colony strength, and heavy winter mortality without any previous evident pathological disturbances. To date there has not been a consensus about its origins. This report describes the clinical features of two professional bee-keepers affecting by this syndrome. Anamnesis, clinical examination and analyses support that the depopulation in both cases was due to the infection by Nosema ceranae (Microsporidia), an emerging pathogen of Apis mellifera. No other significant pathogens or pesticides (neonicotinoids) were detected and the bees had not been foraging in corn or sunflower crops. The treatment with fumagillin avoided the loss of surviving weak colonies. This is the first case report of honeybee colony collapse due to N. ceranae in professional apiaries in field conditions reported worldwide.

Semipreparative chiral supercritical fluid chromatography in the fractionation of lansoprazole and two related antiulcer drugs enantiomers.

The semipreparative chiral separation of lansoprazole and two related compounds (pantoprazole and rabeprazole) using supercritical fluid chromatography (SFC) is presented in this work. Different loads were evaluated in order to obtain high enantiomeric purities and production rates. The volumes injected were 1, 2 and 4 mL. The concentrations of the racemic mixtures were 3 and 6 g/L for lansoprazole and 1.5 g/L for pantoprazole and rabeprazole. In all the cases, the recoveries, for a purity higher than 99.9%, were better for the second eluted enantiomer than for the first one. This fact conditioned the production rate of the first eluted enantiomer that, considering a fixed purity, was always lower than that obtained for the other one. In the case of lansoprazole it was possible to obtain 0.025 and 0.090 mg/min of the first and second eluted enantiomer, respectively, with an enantiomeric purity of 99.9%. For rabeprazole enantiomers 0.037 and 0.062 mg/min, and in the case of pantoprazole the results were better (0.062 and 0.122 mg/min) due to the higher resolution.

Sample preparation methods to analyze fipronil in honey by gas chromatography with electron-capture and mass spectrometric detection.

Several sample preparation methods have been assayed to analyze residues of fipronil in honey. After diluting the honey, liquid-liquid extraction (LLE) with organic solvents, and solid-phase extraction (SPE) on commercial cartridges or Florisil-packed column have been tested at three concentration levels: 0.01, 0.1 and 1 mg/kg. LLE with n-hexane or SPE on a Florisil column resulted to be the most suitable procedures to analyze fipronil at trace concentrations. Fipronil was quantified by gas chromatography (GC) with electron-capture detection (ECD) or mass spectrometric (MS) detection after conventional or matrix-matched calibration. The detection limit of fipronil in honey samples was below 1 microg/kg. The degradation of fipronil in honey has also been studied, being identified three minor degradation products in the extracts.

Use of SPE-GC/EIMS for residue analysis in wine elaborated from musts spiked with formulations of chlorpyriphos-methyl, methiocarb, dicofol, and cyproconazol.

The likely presence in wine of residues of the active ingredient and its degradation products, besides the byproducts and excipients of the commercial formulation, has been investigated for four pesticides. Formulations containing chlorpyriphos-methyl, methiocarb, dicofol, and cyproconazol were added to must, which was subjected to a usual vinification. The wines elaborated from must spiked with the formulation of chlorpyriphos-methyl contained two pyridinol compounds in addition to excipients such as alkylbenzenes, naphthalene, and methylnaphthalenes. Methiocarb was hydrolyzed to yield the corresponding phenol, and various unidentified compounds related to cyproconazol were observed in wine. The residues of the dicofol-containing formulation resulted to be dechlorination products; impurities from its commercial formulation were also detected in must and wine extracts. White wines contained higher amounts of residues than red wines. The residues were detected after an SPE followed by GC/EIMS in the scan mode. The concentrations of the active ingredients were determined by a matrix-matched calibration to avoid quantitative errors arising from the matrix.