A Collaborative Evaluation of LC-MS/MS Based Methods for BMAA Analysis: Soluble Bound BMAA Found to Be an Important Fraction

Exposure to β-N-methylamino-l-alanine (BMAA) might be linked to the incidence of amyotrophic lateral sclerosis, Alzheimer’s disease and Parkinson’s disease. Analytical chemistry plays a crucial role in determining human BMAA exposure and the associated health risk, but the performance of various analytical methods currently employed is rarely compared. A CYANOCOST initiated workshop was organized aimed at training scientists in BMAA analysis, creating mutual understanding and paving the way towards interlaboratory comparison exercises. During this workshop, we tested different methods (extraction followed by derivatization and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis, or directly followed by LC-MS/MS analysis) for trueness and intermediate precision. We adapted three workup methods for the underivatized analysis of animal, brain and cyanobacterial samples. Based on recovery of the internal standard D₃BMAA, the underivatized methods were accurate (mean recovery 80%) and precise (mean relative standard deviation 10%), except for the cyanobacterium Leptolyngbya. However, total BMAA concentrations in the positive controls (cycad seeds) showed higher variation (relative standard deviation 21%–32%), implying that D₃BMAA was not a good indicator for the release of BMAA from bound forms. Significant losses occurred during workup for the derivatized method, resulting in low recovery (<10%). Most BMAA was found in a trichloroacetic acid soluble, bound form and we recommend including this fraction during analysis.     

Floristic diversity of valonia oak silvopastoral woodlands in Greece

Valonia oak (Quercus ithaburensis subsp. macrolepis (Kotschy) Hedge and Yaltirik), a deciduous species grown in the xerothermic conditions of the east Mediterranean zone of Europe, forms woodlands of open canopy that permits adequate light to reach the ground thus sustaining an exceptionally rich flora. In Greece, these woodlands are mainly used for livestock grazing and considered important silvopastoral systems. Nowadays, most of them are abandoned and degraded as a result of inappropriate management. For the effective evaluation and sustainable management of these woodlands and the establishment of reliable monitoring systems, a detailed analysis of their plant diversity characteristics across Greece is needed. This paper aims at determining and comparing floristic diversity measures as well as at assessing the impact of human activities on vegetation and floristic diversity of the most characteristic valonia oak silvopastoral woodlands of Greece. It was found that the truncated log-normal model adequately predicts the species abundance data for eleven out of the twelve woodlands. From the statistical comparison of the diversity indices Q-statistic, Berger–Parker dominance index and Shannon-Wiener measure of equitability, three groups of these woodlands can be distinguished (a) the high-richness group of Pentalofos, Lesvos, Lakonia and Thesprotia, (b) the medium-richness group of Kriti Island, Alexandroupoli, Strofylia and Galaxidi, and (c) the low-richness group of Kea Island, Amfilohia, Almyros and Lilaia. However, the Rényi’s diversity ordering diagram showed that, generally, in low levels of entropy inconsistency is the rule and no clear pattern of diversity is extracted reflecting the impact of complex factors. Among them, those related to management seem to mostly determine plant diversity of valonia oak silvopastoral woodlands, with abiotic factors having an occasionally supporting role. Changes of species richness and evenness due to livestock grazing as well as acorn and wood harvesting are further discussed.     

Gastrointestinal Parasites of Stabled and Grazing Horses in Central and Northern Greece

The aim of this investigation was to compare parasites found in feces from stabled and grazing horses in Greece. For this, a total of 223 fecal samples were collected from horses from various parts of Central and Northern Greece. One hundred fifty were stabled and 73 were grazing horses, used for riding, working, or breeding. One hundred came from seven riding clubs, 73 from one breeding farm, and 50 were work horses from five farms. Parasitologic investigation was performed by applying three fecal examination techniques (Teleman's, zinc flotation, and stained smears) to detect ova, larvae, and oocysts of parasites. It was found that 77 the 223 horses (34.5%) in the study were infected with one or more parasite species. In the stabled horses, the most common parasites detected were eggs of strongyles, Strongyloides spp, Anoplocephala spp, Habronema spp, and Parascaris equorum and oocysts of Eimeria spp and Cryptosporidium spp; in the grazing horses, Anoplocephala spp. Strongyles were significantly more prevalent in the group of stabled horses than in the other group (P < .05). The results of the current study demonstrate that parasite infection is highly prevalent in horses in Greece.     

Attract-and-kill strategy. Laboratory studies on hatched larvae of Culex pipiens

The attract‐and‐kill strategy is a new pest management technique that presupposes the intelligent combination of an attracting agent (e.g. pheromone) and a killing agent (e.g. insecticide). In the present study, the potential combination of the microencapsulated synthetic oviposition pheromone 6‐acetoxy‐5‐hexadecanolide with an insecticide has been tested. Initially, polyurea microcapsules containing 6‐acetoxy‐5‐hexadecanolide, the synthetic mixture of diastereomers of the oviposition pheromone of the mosquito species Culex quinquefasciatus Say (Diptera: Culicidae), were studied. Laboratory bioassays were performed to confirm the bioactivity of the microencapsulated pheromone on the oviposition activity of Culex pipiens L. biotype molestus Førskal (Diptera: Culicidae) with the aim of determining the optimum dose for oviposition response. Its effect was dose dependent, revealing an optimum dose of 300 mg of dried microcapsules. Attractancy over time was also studied. The microencapsulated pheromone was found to be sufficiently attractive to gravid female mosquitoes for a period of 40 days. Finally, the combination of the synthetic pheromone with the control agent temephos showed both an acceptable oviposition activity and sufficient larvicidal effect. Copyright © 2007 Society of Chemical Industry     

A wild blueberry-enriched diet (Vaccinium angustifolium) improves vascular tone in the adult spontaneously hypertensive rat

The effect of a wild blueberry-enriched diet on vasoconstriction and vasorelaxation was examined in the adult, 20-week-old spontaneously hypertensive rat (SHR) after 8 weeks of a control (C) or an 8% wild blueberry (WB) diet. Nitric oxide (NO)- and cyclooxygenase (COX)-mediated aortic responses were examined ex vivo with the agonists L-phenylephrine (Phe) and acetylcholine (Ach), in the absence or presence of the NO synthase (NOS) inhibitor L-NG-monomethyl arginine (L-NMMA) or the COX inhibitor mefenamic acid (MFA). The vasoconstriction elicited by Phe was reduced in the WB group, attributed to the NO pathway, favoring a lower vascular tone under basal conditions. Acetylcholine-induced vasorelaxation in the WB group was possibly mediated through the COX, but not the NO pathway. These findings document the potential of wild blueberries to modify major pathways of vasomotor control and improve the vascular tone in the adult SHR with endothelial dysfunction.     

Composition and antioxidant activity of the essential oils of Xylopia aethiopica (Dun) A. Rich. (Annonaceae) leaves, stem bark, root bark, and fresh and dried fruits, growing in Ghana

The chemical composition of the essential oils obtained from the leaves, the barks of the stem and the root, as well as from the fresh and dried fruits of Xylopia aethiopica, growing in Ghana, was investigated by gas chromatography/mass spectrometry analyses. Kovats indices, mass spectra, and standard compounds were used to identify a total of 93 individual compounds. The monoterpene hydrocarbons formed the main portion in all studied samples. beta-Pinene was predominant in all cases, while trans-m-mentha-1(7),8-diene was the main compound in the essential oils of the leaves and the barks of roots and stems. Their potential antioxidant activity was also investigated and found to be significant in scavenging superoxide anion radical.     

Effect of solution conductivity on the volatile constituents of Origanum dictamnus L. in nutrient film culture

The chemical composition of the essential oils obtained from leaves and bracts of hydroponically cultivated Origanum dictamnus L. (Cretan dittany), growing under various electrical conductivity (EC) levels (2.0, 4.0, and 6.0 mS/cm), was studied, using the nutrient film technique (NFT). The analysis of the essential oil content was achieved by GC-MS technique, and totals of 41 and 38 different compounds were identified in both cases of large-leaved and narrow-leaved samples of leaves and bracts, respectively. Differences in the composition content and of the percentage of each of the constituents in the two studied samples (i.e., large-leaved and narrow-leaved) and within the essential oils of leaves and bracts in both samples were observed. Carvacrol and p-cymene were identified as the main constituents in all essential oils, whereas thymoquinone was found in higher percentage in the essential oils of large-leaved than in narrow-leaved plants. The results obtained from GC-MS analysis were submitted to chemometric analysis, and a phenotypic similarity of the essential oils of narrow-leaved O. dictamnus was observed, whereas the essential oils of large-leaved O. dictamnus showed two separate subgroups.