Application of isotope dilution analysis for the evaluation of extraction conditions in the determination of total selenium and selenomethionine in yeast-based nutritional supplements

Isotope dilution analysis (IDA) has been used to quantify total selenium, total solubilized selenium, and the selenomethionine (SeMet) amount in yeast and yeast-based nutritional supplements after acid microwave digestion and different enzymatic extraction procedures. For this purpose, both a (77)Se-enriched SeMet spike, previously synthesized and characterized in our laboratory, and a (77)Se(VI) spike were used. In the analysis of the nutritional supplements, the SeMet spike was added to the sample and extracted under different conditions, and the (78)Se/(77)Se and (80)Se/(77)Se isotope ratios were measured as peak area ratios after high-performance liquid chromatography (HPLC) separation and inductively coupled plasma mass spectrometry (ICP-MS) detection. The formation of SeH(+) and mass discrimination were corrected using a natural SeMet standard injected every three samples. Similarly, total solubilized selenium was measured in the extracts after enzymatic hydrolysis using the (77)Se-enriched SeMet as a spike by direct nebulization without a chromatographic separation. To establish a mass balance, total selenium was also determined by IDA-ICP-MS on the yeast tablets after microwave digestion using (77)Se(VI) as a spike. Results showed that all enzymatic procedures tested were able to solubilize total selenium quantitatively from the solid. However, the recovery for the species SeMet, the major selenium compound detected, was seriously affected by the enzymatic procedure employed and also by the matrix composition of the supplement evaluated. For the yeast sample, SeMet recovery increased from 68 to 76% by the combined use of driselase and protease. For the nutritional supplements, the two most effective procedures appeared to be protease and driselase/protease, with a SeMet recovery ranging from 49 to 63%, depending upon the supplement evaluated. In the case of in vitro gastrointestinal enzymolysis, the results obtained showed 26-37% SeMet recovery, while the rest of selenium was solubilized as other unknown compounds (probably Se-containing peptides).     

Spice crops agroforestry systems in the East Usambara Mountains,Tanzania: growth analysis

A scarcity of cultivation land calls for more intensive and productive land use in the East Usambara Mountains in NE Tanzania. Spice crops could generate cash in higher parts of the mountains, but the present cultivation methods are depleting the valuable forest resources. The trial was established at the end of 2000 to find out how the two popular cash crops, cardamom (Elettaria cardamomum (L.) Maton.) and black pepper (Piper nigrum L.), normally grown under the natural forest, will produce in intensive agroforestry system with two multipurpose farm trees, Grevillea robusta A.Cunn. and nitrogen fixing Gliricidia sepium Jacq. Results from 6 years showed that cardamom produced better with grevillea than in natural forest; 5.5 times more in the fourth year than the average in the area. The Land Equivalent Ratios for black pepper and cardamom showed that pepper intercropped with grevillea produced 3.9 times more than in monoculture whereas cardamom intercropped with grevillea and pepper produced 2.3 times more than in monoculture. Gliricidia improved the nitrogen and organic matter content of the soil over the levels found in natural forest. Soil acidity was, however, preventing the plants from using the available mineral nutrients more effectively.     

Analysis of Quality Protein Changes in Nixtamalized QPM Flours as a Function of the Steeping Time

This study showed the protein changes in Quality Protein Maize (QPM H‐368C) during the traditional nixtamalization process as a function of the steeping time from 0 to 15 hr. Protein content (N × 6.25), pH, protein fractionation, reactive lysine, essential amino acids, and protein digestibility were analyzed to explain the protein quality modifications in nixtamalized corn flours (NQF). The thermoalkaline process increased significantly (P ≤ 0.05) the protein content (5.57 ± 0.86%) in NQF obtained at 3, 5, 7, 9, 11, 13, and 15 hr of steeping time compared with native corn or corn without treatment (NC). The pH values of NQF were not proportional to the steeping time and significantly different (P ≤ 0.05) between them. At 5 hr critical steeping time, the total lysine and reactive lysine content decreased severely (36 and 32%, respectively) with statistical differences (P ≤ 0.05) compared with NC. On the other hand, the tryptophan content decreased significantly (P ≤ 0.05) at steeping times of 5–15 hr (38.70 ± 6.7%) compared with NC. The changes in the lysine and tryptophan content were not proportional to the steeping time. The protein recovery in the albumin and globulin fraction diminished (P ≤ 0.05) with respect to raw corn. The protein recovery for γ‐zeins, glutelin‐like proteins, glutelins, and residue increased. A significant (P ≤ 0.05) decrease was found in the essential amino acids in NQF with 3–7 hr of steeping time compared with NC. Equally important was the reduction in protein digestibility observed in NQF steeped at long steeping times (11–15 hr) with significant (P ≤ 0.05) differences compared with NC. The protein solubility distribution along the steeping step and the essential amino acids location, specifically lysine in corn kernel, could explain partially the protein quality changes observed in this research. Finally, these results contribute to reconciling discrepancies associated with the protein quality modifications in nixtamalized corn reported previously in literature.     

Resistance monitoring in codling moth: a need for standardization

BACKGROUND: A wide range of methods has been used for monitoring resistance in Cydia pomonella L. However, the results from specific tests are not always concordant and they generate doubts over the extent to which the data represent the field situation. Therefore, the variability in the expression of resistance mechanisms and in resistance ratios between developmental stages and their reliability in a field situation were studied. RESULTS: Neonate larvae showed the highest insecticide resistance, except to spinosad. Resistance ratios were from 2 to 9.5, depending on the insecticide and the strain tested, when compared with the susceptible strain. Spinosad exhibited the highest resistance ratio for diapausing larvae (4.4, 12.2 and 4.4 for Raz, Rdfb and RΔ strains, respectively). Enzymatic ratios were higher in adults than in neonates or diapausing larvae, and the highest enzymatic ratios were observed for mixed‐function oxidases (7.3–36.7 for adults of resistant strains). CONCLUSIONS: The sensitivity of different instars depends on the insecticides used, and there is a differential expression of resistance mechanisms between stages. For the population considered in this study, tests on neonates provided the most consistent results to the field situation, but adult response to chlorpyrifos‐ethyl and azinphos‐methyl in the laboratory was also consistent with the field results. Copyright © 2008 Society of Chemical Industry     

Geographic variation in pheromone component ratio and antennal responses,but not in attraction,to sex pheromones among fall armyworm populations infesting corn in Mexico

The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), is one of the most important pests of corn in Latin America. This insect presents two strains with behavioural and genetic differences. In Mexico, both strains are present, and at least two different FAW populations have been reported within the corn-strain. The objective of this study was to gather evidence of whether pheromonal communication varies among different S. frugiperda populations infesting corn in Mexico. First, we investigated any qualitative or quantitative difference in the composition of sex pheromones among populations; second, we studied whether male antennal responses to pheromone components vary among populations; and, finally, we investigated whether males from a region can discriminate between a synthetic pheromone blend characteristic of their region and blends formulated with the ratio of pheromone compounds emitted by females from other populations. Sex pheromone components were sampled by solid-phase microextraction and identified by gas chromatography coupled to mass spectrometry. Females from all populations consistently released three compounds: (Z)-9-tetradecenyl acetate (Z9-14:OAc), (Z)-11-hexadecenyl acetate (Z11-16:OAc) and (Z)-7-dodecenyl acetate (Z7-12:OAc). Our results showed that Z9-14:OAc and Z7-12:OAc evoked the highest and most variable antennal responses among the populations studied compared to Z11-16:OAc. However, males did not discriminate between local pheromone blends and those formulated from other populations in a field test. These results show that although there is geographic variation in the ratio of pheromone components and in the peripheral reception of them, males were not differentially attracted to different pheromone blends in the field.     

Tree effects on the chemical topsoil features of oak, beech and pine forests

We aimed to study tree effects on the chemical properties of forest soils. We compared soil features of three types of forest ecosystems, each with four stands (replicates): beech forests (Fagus sylvatica), oak forests (dominated by Quercus pyrenaica) and pine plantations (Pinus sylvestris). Five samples from the top 10 cm of soil were taken per stand, from which pH, organic matter content (O.M.), total nitrogen (N) and available calcium (Ca²⁺), magnesium (Mg²⁺), potassium (K⁺) and sodium (Na⁺) were determined. Litter layer depth was measured at each soil sampling point. We also measured tree density and crown diameters at each stand. Our results indicated that soil samples from the four pine plantation stands were more similar while oak and beech stands were characterised by great variability in terms of soil properties and leaf litter depth. Although the identity of the dominant tree species significantly influenced several topsoil chemical properties (increase in pH and available cations in oak forests and higher organic matter and total nitrogen in beech and pine ecosystems), there were other important factors affecting soil features that may be taken under consideration. Differences between soil properties of the three types of forest ecosystems were mainly related to the characteristics of the litter layer and less related to the tree layer structure. Finally, the establishment of pine plantations in naturally deciduous tree areas made the topsoil features more homogeneous.     

Evidence that active transmission of porcine cysticercosis occurs in Venezuela

There is a paucity of quantitative data on the status of porcine cysticercosis in Venezuela, information which is essential for understanding the level of disease transmission. This study was, therefore, conducted in a typical small rural community in Yaracuy State, Venezuela, where previous cases of human Taenia solium taeniasis/cysticercosis had been reported and where the free-ranging pig management practices and the lack of rudimentary sanitary facilities indicated an obvious risk for transmission of the disease. Serum samples from 52 village pigs were screened by enzyme-linked immunosorbent assays for anti-cysticercal antibodies (Ab-ELISA), using T. solium cyst fluid as the antigen and the HP10, monoclonal antibody-based, antigen trapping ELISA for parasite antigen (HP10 Ag-ELISA). Significantly, a high proportion of the animals (65.4% for the Ab-ELISA and 42.3% for the HP10 Ag-ELISA) were sero-positive. Five of the pigs, which were selected on that basis of positive tongue palpation, were killed for autopsy, and large numbers of viable cysticerci were found in the carcases. This unequivocal documentation of porcine cysticercosis in Venezuelan pigs presents clear evidence that T. solium is actively transmitted in Venezuela. Further detailed studies and implementation of appropriate control measures are therefore indicated.     

Expression of vascular endothelial growth factor and endothelial nitric oxide synthase is increased in the placenta of sheep at high altitude in the Andes

Fetal weight and the placenta of sheep at high altitude (HA) are affected by hypoxia. Placental changes (an increase in placental size and vascularization) are greater in ewes from populations that have lived for several generations at HA than in those exposed during just 1 gestation. This study investigated placental expression of vascular endothelial growth factor (VEGF) and endothelial nitric oxide synthase (eNOS), 2 molecules involved in placental angiogenesis that could be upregulated by hypoxia. Two groups of ewes were maintained at HA (3589 m) during pregnancy: HA-native ewes (group HH) and ewes native to lowlands but moved to HA immediately after the diagnosis of pregnancy (group LH). A control group (LL) was kept at sea level. Near term, placentomes were removed, weighed, and processed for immunohistochemical detection of VEGF and eNOS, as well as for vascular area measurement. Placental weight was significantly higher in the HH group than in the LH and LL groups; between the latter 2 groups there was no significant difference. The placental area occupied by vasculature was significantly greater in both the HA groups than in the LH group; the number of placentomes was greatest in the LL group. The density of VEGF and eNOS in the placentome tissue was significantly greater in both HA groups than in the LL group. Although the density of VEGF was significantly lower in the HH group than in the LH group, no differences were observed in eNOS density between the HH and LH animals. These results demonstrate that chronic hypoxia upregulates the expression of placental VEGF and eNOS, suggesting an important role of these molecules in the placental response to HA hypoxia. In addition, an attenuated response to hypoxia in VEGF synthesis may be part of the long-term process of adaptation to HA.     

d24-hour changes in circulating prolactin, follicle-stimulating hormone, luteinizing hormone and testosterone in male rats subjected to social isolation

BACKGROUND: This work analyzes the effect of social isolation (a mild stressor) on the 24-h variation of pituitary-testicular function in young Wistar rats, assessed by measuring circulating levels of prolactin, FSH, LH and testosterone. METHODS: Animals were either individually caged or kept in groups (4-5 animals per cage) under a 12:12 h light-dark cycle (lights on at 0800 h) for 30 days starting on day 35 of life. Rats were killed at 4-h intervals during a 24-h cycle, beginning at 0900 h. RESULTS: Isolation brought about a decrease in prolactin, LH and testosterone secretion and an increase of FSH secretion. In isolated rats the 24-h secretory pattern of prolactin and testosterone became modified, i.e., the maximum in prolactin seen in control animals at the beginning of the activity span was no longer detected, whereas the maximum in circulating testosterone taking place at 1700 h in controls was phase-delayed to 2100 h in isolated rats. CONCLUSION: Social isolation affects the 24-h variation of pituitary-testicular function in young rats. Secretion of prolactin, LH and testosterone decreases, and secretion of FSH increases, in isolated rats. The maximum in prolactin seen in group-caged rats at the beginning of the activity span is not observed in isolated rats. The maximum in circulating testosterone taking place at the second part of the rest span in controls is phase-delayed to the light-dark transition in isolated rats.