Further studies on the analysis of DSP toxin profiles in galician mussels

Further studies on mussel samples from Galicia, Spain, have revealed the presence of okadaic acid (OA), dinophysistoxin 2 (DTX2), and the fatty acid acyl esters of both of these toxins as the "DTX3" complex. Measurements were performed with an improved in situ method for the formation of 9-anthryldiazomethane (ADAM) derivatives followed by liquid chromatography with fluorescence detection. Base hydrolysis of DTX3 toxins gave free OA and DTX2, which were determined following ADAM derivatization. Results were confirmed by liquid chromatography/mass spectrometry analyses, and in most of the samples, free DTX2 was the most abundant toxin. However, the OA/DTX2 ratio in the DTX3 conjugated form was different, with OA being the most abundant in all cases. This difference could be due to different rates of metabolism of OA and DTX2 to the acyl esters or due to contamination of the shellfish by the two toxins at different points in time, resulting in less acyl ester formation for one toxin versus the other. The second possibility would be reasonable if two different source organisms were producing the toxins.     

Data analysis for molecular characterization of plant genetic resources

Characterizations of plant genetic resources based on molecular markers have been increased in the last years. Studies using a broad range of markers applied on hundreds of plant species are the theoretical basis for inferring genetic diversity to propose both breeding and conservation strategies. Despite increased importance of molecular characterization in plant genetic resources, there is scarce information about analysis of this type of data. To fill this gap of information, this review discuss the rationale behind analyses achieved to study genetic relationship among accessions (within and between groups) and to identify accession, and also discuss the adequacy of some analyses and/or parameters for specific purposes. Genetic diversity within groups may be either quantified for the whole group (parameters to choose will depend on type of marker), or quantified and visualized for the relationships among individuals. Quantification parameters will be chosen depending on type of marker, reproduction mode and relatedness of individuals. Visualization is achieved by hierarchical and non-hierarchical methods. Genetic diversity between groups should be quantified either by analysis of molecular variance, or Nei’s parameters, or Wright’s F-statistics. Efficiency of accession identification can be evaluated by maximal probability of identical match by chance and number of resolved genotypes.     

Functional markers delimiting a <Emphasis Type="Italic">Medicago</Emphasis> orthologue of pea symbiotic gene <Emphasis Type="Italic">NOD3</Emphasis>

Symbiotic gene mutated in the pea (Pisum sativum L.) line RisfixC is a determinant of the number of symbiotic root nodules. In parallel to a sharp increase in nodule number, its mutational inactivation brings about the insensitivity of nodulation to the ambient nitrate level (Nts trait). Using the established localization to the SYM2-NOD3 region of the pea linkage group I, functional PCR markers were developed for the orthologous region on the chromosome 5 of the model species Medicago truncatula. Owing to the conservation of the binding regions of the designed primers, pea orthologues were successfully amplified with 60% of the primer pairs tested. When applied to a mapping pea population from the cross of the line RisfixC x Afghanistan L1268 (sym2), the new markers allowed to localize the supernodulation mutation within 2.5 cM confidence interval in the pea genome. The placement of the functional markers on the M. truncatula chromosome 5 confined the orthologous gene location to eight overlapping BACs spanning approximately 710 kbp (positions 37,755,678–38,467,472). The narrowed list of the annotated Medicago genes in combination with the published data on their symbiotic and nitrate regulation can be used for the candidate gene identification, together with the requirements imposed by the known function in nodule number initiation and nitrate sensing. In addition, the new markers are applicable for tracking the RisfixC allele in breeding programmes aimed at the improvement of symbiotic performance.     

Stress responses in adult cattle due to surgical dehorning using three different types of anaesthesia

Dehorning adult cattle is a surgical procedure causing distress of varying intensities that can be reflected in behavioural changes and alterations in plasma cortisol levels. Stress responses during the dehorning process were evaluated in 18 Red Pied cows. The cows were divided into 3 groups of six and kept in tie-stall housing. Those in the first group were dehorned under general anaesthesia (GA) induced by intravenous administration of xylazine and ketamine. The second group was dehorned under sedation and local anaesthesia (SLA) induced by intramuscular administration of xylazine and local anaesthesia with lidocaine. The third group was dehorned under local anaesthesia (LA) with lidocaine. Dehorning was performed with a foetotomy wire. Blood samples were taken 0.5 h before dehorning to determine cortisol levels, and, by means of a central venous catheter inserted into the jugular vein, during surgery at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7 and 8 h post-surgery. Concurrently, occurrence of stress behaviours was assessed. Cortisol levels were measured by enzyme immunoassay (EIA). The lowest mean peak levels of plasma cortisol (82.53 +/- 6.04 nmol l(-1)), the most rapid return of plasma cortisol levels to baseline values (1.92 +/- 1.11 h), and the lowest occurrence of stress behaviours (2.38 +/- 5.83%) were noted in the SLA group. The highest mean peak levels plasma cortisol (113.86 +/- 25.65 nmol l(-1)), the slowest return of plasma cortisol levels to baseline values (3.83 +/- 2.18 h) and the most frequent occurrence of stress behaviours (65.48 +/- 28.72%) were observed in the LA group. There were significant differences between the SLA and LA groups in peak plasma cortisol levels (p = 0.011) and in occurrence of stress behaviours (p = 0.003). Sedation induced by intramuscular administration of xylazine in conjunction with local anaesthesia with lidocaine is considered the most suitable method of anaesthesia when dehorning adult cattle. Local anaesthesia with lidocaine alone was least suitable.     

Intestinal antiinflammatory activity of a lyophilized infusion of Turnera ulmifolia in TNBS rat colitis

Turnera ulmifolia is a plant popularly known in Brazil and South America as chanana. Some species of Turnera are widely used in folk medicine for different types of inflammatory diseases. In this study, the preventive intestinal antiinflammatory activity of a lyophilized infusion obtained from the aerial parts of T. ulmifolia was tested in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. The results obtained revealed that pretreatment to colitic rats with the extract, at 250 and 500 mg/kg, significantly attenuated the colonic damage induced by TNBS. This beneficial effect was associated with an improvement in the colonic oxidative status, since the infusion prevented the glutathione depletion that occurred as a consequence of the colonic inflammation. On the other hand, this antioxidant activity was confirmed in in vitro studies. In conclusion, the preventive effect exerted by the lyophilized infusion of T. ulmifolia in the TNBS model of rat colitis is probably related to its antioxidant properties, due to its flavonoids content.     

Hypolipidemic effect in cholesterol-fed rats of a soluble fiber-rich product obtained from cocoa husks

A new soluble cocoa fiber product (SCFP), obtained after enzymatic treatment of cocoa husks, was characterized and its potential health effects studied in an animal model of dietary-induced hypercholesterolemia. The SCFP was rich in soluble dietary fiber (DF) and antioxidant polyphenols. Consumption of a cholesterol-rich diet containing the SCFP as a source of DF resulted in lower food intake and body weight gain in comparison with control groups consuming cholesterol-free or cholesterol-rich diets with cellulose as DF. The cholesterol-rich diet caused remarkable hypercholesterolemia. However, the SCFP diminished the negative impact of the cholesterol-rich diet, buffering the decrease of high density lipoprotein-cholesterol, and the increase of total and low density lipoprotein-cholesterol levels, and lipid peroxidation (malondialdehyde levels) induced by the fatty diet. The SCFP also decreased triglyceride levels to values lower than those in the group fed the cholesterol-free diet. These results put forward the potential application of the SCFP as a dietary supplement or functional food ingredient.     

Chemical composition of Lippia spp. essential oil and antimicrobial activity against Aeromonas hydrophila

Phytotherapy can replace antibiotic administration as an alternative to control Aeromonas hydrophila, one of the main bacteria involved in the aetiology of farmed fish diseases. Given that plants of the Lippia spp. genus show biological potential for antimicrobial activity, this study evaluated the chemical composition of essential oils extracted from Lippia alba, Lippia origanoides and Lippia sidoides and their activity against A. hydrophila. The oils were obtained by steam distillation in a Clevenger‐type apparatus and their composition determined by gas chromatography and mass spectrometry (CG/MS). Antibacterial activity was assessed by calculating the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using broth microdilution method. The main compounds identified were geranial (25.4%) and neral (16.6%) in L. alba oil, carvacrol (40.4%) and p‐cymene (11.4%) in L. origanoides oil and thymol (76.6%) and ortho‐cymene (6.3%) in L. sidoides oil. The three Lippia species showed bacteriostatic and bactericidal action against A. hydrophila, with MICs and MBCs ranging from 1250 to 5000 μg mL^?1. Of the species tested, the best performance was obtained with essential oil of L. sidoides.