Arsenic-contaminated soils phytotoxicity studies with sunflower and sorghum

Background, Aim and Scope  Environmental pollution caused by arsenic (As) is a major ecological problem. There has been intense worldwide effort to find As-hyperaccumulating plants that can be used in phytoremediation—the green-plant-assisted removal of chemical pollutants from soils. For phytoremediation, it is natural to prefer cultivated rather than wild plants, because their agriculture is well known. This study was conducted to evaluate the tolerance of common sunflower(Helianthus annuus L.) and sugar sorghum(Sorghum saccharatum Pers.) for soil-As contents of 10–100 mg As kg^-1 soil, with sodium arsenite as a model contaminant. Methods  Plants were grown in a growth chamber for 30 days. Microfield experiments were conducted on experimental plots. To study the phytoremediation effect of the auxins indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), we treated 1- and 3-day-old plant seedlings with water solutions of the auxins (concentrations 10^-5, 10^-7, and 10^-9 g l^-1). The soil and plant-biomass samples were analyzed for total As by using the color reaction of ammonium molybdate with As. Results and Discussion  Phytotoxicity studies showed that 100 mg As kg^-1 soil poisoned sunflower and sorghum growth by 50%. There was a linear correlation between soil-As content and As accumulation in the plants. Laboratory experiments showed that the soil-As content was reduced two- to threefold after sunflower had been grown with 10–100 mg As kg^-1 soil for 30 days. Treatment of sunflower and sorghum seedlings with IAA and 2,4-D at a concentration of 10^-5 g l^-1 in microfield experiments enhanced the phytoremediation two- to fivefold as compared with untreated control plants. The best results were obtained with 3-day-old seedlings. Conclusion, Recommendation and Outlook  (a) Sunflower and sorghum are good candidates to remediate As-polluted soils. (b) Phytoremediation can be improved with IAA or 2,4-D. (c) Mixed cropping of sorghum and sunflower may be another way of improving phytoremediation.     

Effects of a 10-year conservation programme on the genetic diversity of the Pottoka pony--new clues regarding their origin

Here, we present the results of a genetic analysis of 463 Pottoka ponies corresponding to four generations, using 17 microsatellite markers. Ten years after the beginning of the Pottoka conservation programme, the values for the genetic diversity of the breed are still high and stable, indicating the success of the programme. We found null alleles in Pottoka for the ASB23, HMS3 and HTG10 microsatellites. Together with information obtained from other pony breeds from the Iberian Peninsula, this finding indicates that these microsatellites should not be used for phylogenetic analyses or parentage tests, at least for these breeds. The high heterozygosity exhibited by this breed in comparison to other ponies, together with its genetic proximity to the centroid of the allele frequencies, suggest that Pottoka allele frequencies are close to those initially exhibited by the ancestors of current European ponies. The results obtained in the current work, together with results from previous studies of ponies and horses from the Iberian Peninsula, corroborate the idea of a unique origin of all ponies from the European Atlantic Area. In contrast, our results do not corroborate the idea that these are derived from a domestication event in the Iberian Peninsula, nor that they have incorporated ancient Iberian horse genes into their genetic pool to a larger extent than other horse breeds.     

The fate of arsenic in soils adjacent to an old mine site (Bustarviejo,Spain): mobility and transfer to native flora

Background, aim, and scope  

The mobility of arsenic in soils and its transfer to other environmental components present significant environmental risks. The management of polluted land is determined by the availability, mobility, and transfer of inorganic pollutants to different ecosystem compartments. In this paper, the fate of arsenic at this mining site has been evaluated to determine future management practises to minimise such risk.     

Plant extracts, spices, and essential oils inactivate Escherichia coli O157:H7 and reduce formation of potentially carcinogenic heterocyclic amines in cooked beef patties

Meats need to be heated to inactivate foodborne pathogens such as Escherichia coli O157:H7. High-temperature treatment used to prepare well-done meats increases the formation of carcinogenic heterocyclic amines (HCAs). We evaluated the ability of plant extracts, spices, and essential oils to simultaneously inactivate E. coli O157:H7 and suppress HCA formation in heated hamburger patties. Ground beef with added antimicrobials was inoculated with E. coli O157:H7 (10(7) CFU/g). Patties were cooked to reach 45 °C at the geometric center, flipped, and cooked for 5 min. Samples were then taken for microbiological and mass spectrometry analysis of HCAs. Some compounds were inhibitory only against E. coli or HCA formation, while some others inhibited both. Addition of 5% olive or apple skin extracts reduced E. coli O157:H7 populations to below the detection limit and by 1.6 log CFU/g, respectively. Similarly, 1% lemongrass oil reduced E. coli O157:H7 to below detection limits, while clove bud oil reduced the pathogen by 1.6 log CFU/g. The major heterocyclic amines 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were concurrently reduced with the addition of olive extract by 79.5% and 84.3% and with apple extract by 76.1% and 82.1%, respectively. Similar results were observed with clove bud oil: MeIQx and PhIP were reduced by 35% and 52.1%, respectively. Addition of onion powder decreased formation of PhIP by 94.3%. These results suggest that edible natural plant compounds have the potential to prevent foodborne infections as well as carcinogenesis in humans consuming heat-processed meat products.     

Efficiency of Mesocosm-Scale Constructed Wetland Systems for Treatment of Sanitary Wastewater Under Tropical Conditions

Subsurface-flow constructed wetlands technology (SSFW) has been used successfully for treating sanitary wastewater throughout North America and Europe. However, treatment wetland technologies have not been used extensively in the tropics. To advance tropical studies, a pilot-scale SSFW was constructed on the campus of the University of the Atlantic in Barranquilla, Colombia. The systems performance was monitored from January to July of 2009. The treatment system consisted of a 760-L septic tank followed by three mesocsom-scale subsurface-flow constructed wetlands in parallel arrangement. Clarified wastewater was batch loaded to each unit at a rate of 53 L/m²/day to affect a hydraulic retention time of approximately 3 days. One of the treatment units served as a non-planted control (gravel only), while the other two treatment units were planted with either Eriochloa aristata or Eleocharis mutata. The objective of this study was to evaluate the comparative efficacy of the treatment units (planted vs. unplanted), with respect to their abilities to augment treatment of septic tank effluent (sanitary wastewater). Monitored parameters included plant biomass, oxidation?Creduction potential, chemical oxygen demand (COD), temperature, dissolved oxygen, pH, ammonia?Cnitrogen (NH ₄ ⁺ ?CN) nitrate?C and nitrite?Cnitrogen (NO₃?CN, NO₂?CN), phosphates (PO ₄ ^? ), and coliform bacteria. Total biomass (dry matter) was 2.84 and 0.87 Kg/m² for E. aristata and E. mutata, respectively. Redox potential in the plant rizospheres averaged ?172 mV (±164.1) in E. aristata, 29 mV (±251.1) in E. mutata, and 32 mV (±210.5) in the unplanted control. COD removal was superior in planted vs. non-planted systems (>75% vs. 47%). Ammonia and total phosphorus removal averaged 69% and 85%, respectively, in planted systems versus 31% and 59% in the unplanted system. Removal of total and fecal coliforms averaged 96%. Results of this pilot study revealed that SSFW technology in the tropics can provide significant removal of organic matter, nutrients, and bacteria from clarified sanitary wastewater.     

Daily and Annual Variation in Digestive Enzymes – Amylase and Basic and Acid Proteases – in Gilt‐head Sea Bream,Sparus aurata

This study examines the daily and annual changes in amylase and basic and acid protease activities in gilt‐head sea bream, Sparus aurata. During the experiment, the fish were kept in cages for production under natural temperature and photoperiod conditions and fed with a commercial diet. To study the effect of photoperiod and temperature, the experiments were conducted during six different months that were chosen based on temperature and photoperiod. To determine daily changes in enzymes activities, for every 3 h during 24 h period, six fish were randomly selected and sacrificed. The annual results for acid protease activity show a rhythm with 6‐mo period (bimodal rhythm) with acrophases in May and November. The activities of pancreatic enzymes, which include basic protease and amylase, exhibited parallel changes with two peaks of activity in January and October without a rhythmic pattern. The daily changes in enzyme activities are significant only in May, June, and November for basic proteases and May, October, and November for amylase (P < 0.05). The observed variations suggest a varying capacity for digestion depending on the day and the year and could aid in the study of seasonal feeding protocols.     

Purification, N-terminal sequencing and diagnostic value of the major antigens of Ornithodoros erraticus and O. moubata

To enhance the specificity and sensitivity of serological detection of swine exposed to Ornithodoros erraticus or O. moubata, we purified the 158, 186, 215 and 260 kDa antigens from the former species and the designated (owing to their MW and charge) 19C, 17A, 20A1 and 20A2 antigens of the latter by HPLC and gel electroelution methods. All the O. erraticus antigens share epitopes and are difficult to purify individually by reverse phase and ion-exchange chromatography due to their molecular similarity. Tested individually by ELISA, all of them give the same optical densities (OD) with anti-O. erraticus sera, and these ODs are always lower with anti-immature than with anti-adult sera. Although immature and adult specimens have the same antigens, immature forms induce more anti-carbohydrate antibodies than adults. This is the reason for the lower ODs of the anti-immature sera against purified antigens, since these latter antigens essentially react with anti-peptide antibodies (hence, increasing the specificity and sensitivity of the serology). The N-terminus of the 260 kDa antigen shows 80-90% similarity with the hemoglobin alpha-chain of many mammals. The antigens of O. moubata are proteins that are very different from one another and are, therefore, readily purified by ion exchange chromatography. The 20A1 antigen appears to be the most immunogenic and is recognized equally by anti-immature and anti-adult sera. This antigen does not give false positive reactions with the negative control sera analyzed and its N-terminus region shares 46.2% homology with the alpha-chain of the C3 component of rabbit complement.     

Screening of postharvest agricultural wastes as alternative sources of peroxidases: characterization and kinetics of a novel peroxidase from lentil ( Lens culinaris L.) stubble

Aqueous crude extracts of a series of plant wastes (agricultural, wild plants, residues from sports activities (grass), ornamental residues (gardens)) from 17 different plant species representative of the typical biodiversity of the Iberian peninsula were investigated as new sources of peroxidases (EC 1.11.1.7). Of these, lentil (Lens culinaris L.) stubble crude extract was seen to provide one of the highest specific peroxidase activities, catalyzing the oxidation of guaiacol in the presence of hydrogen peroxide to tetraguaiacol, and was used for further studies. For the optimum extraction conditions found, the peroxidase activity in this crude extract (110 U mL(-1)) did not vary for at least 15 months when stored at 4 °C (k(inact) = 0.146 year(-1), t(1/2 inact) = 4.75 year), whereas, for comparative purposes, the peroxidase activity (60 U mL(-1)) of horseradish (Armoracia rusticana L.) root crude extract, obtained and stored under the same conditions, showed much faster inactivation kinetics (k(inact) = 2.2 × 10(-3) day(-1), t(1/2 inact) = 315 days). Using guaiacol as an H donor and a universal buffer (see above), all crude extract samples exhibited the highest peroxidase activity in the pH range between 4 and 7. Once semipurified by passing the crude extract through hydrophobic chromatography on phenyl-Sepharose CL-4B, the novel peroxidase (LSP) was characterized as having a purity number (RZ) of 2.5 and three SDS-PAGE electrophoretic bands corresponding to molecular masses of 52, 35, and 18 kDa. The steady-state kinetic study carried out on the H(2)O(2)-mediated oxidation of guaiacol by the catalytic action of this partially purified peroxidase pointed to apparent Michaelian kinetic behavior (K(m)(appH(2)O(2)) = 1.87 mM; V(max)(appH(2)O(2)) = 6.4 mM min(-1); K(m)(app guaicol) = 32 mM; V(max)(app guaicol) = 9.1 mM min(-1)), compatible with the two-substrate ping-pong mechanism generally accepted for peroxidases. Finally, after the effectiveness of the crude extracts of LSP in oxidizing and removing from solution a series of last-generation dyes present in effluents from textile industries (1) had been checked, a steady-state kinetic study of the H(2)O(2)-mediated oxidation and decolorization of Green Domalan BL by the catalytic action of the lentil stubble extract was carried out, with the observation of the same apparent Michaelian kinetic behavior (K(m)(appGD) = 471 μM; V(max)(appGD)= 23 μM min(-1)). Further studies are currently under way to address the application of this LSP crude extract for the clinical and biochemical analysis of biomarkers.     

Quantification of Bacillus thuringiensis Cry1Ab protein in tissues of YieldGard (MON810) corn hybrids tested at multiple field locations in India

Transgenically expressed Bacillus thuringiensis insecticidal-protein Cry1Ab was quantified in target tissues of insect feeding of several YieldGard^® corn hybrids. The Cry1Ab protein is intended to protect corn plants from two economically important stem borers, Chilo partellus and Sesamia inferens. A total of seven YieldGard hybrids, all with MON810 event, were field-tested in a total of fourteen locations during the dry season (October–March) of 2005/2006 and wet season (May–October) of 2006. S. inferens and C. partellus oviposit on leaves of young corn plants, 15–60 days after emergence (DAE). The neonates initially feed by scraping the leaf lamina before migrating to bore into the stem. Thus high concentrations of Cry1Ab in whorl leaf and stem tissues would ensure effective control of the borers. The mean tissue Cry1Ab concentrations during the oviposition window of the borers (15–60 DAE), ranged from 50.05 to 21.01 ppm in whorl leaf, and between 9.26 and 3.47 ppm in stem tissue during the same period in the dry season of 2005/06. Similarly, Cry1Ab concentrations in whorl leaf and stem between 15 and 60 DAE during the wet season of 2006 ranged between 19.30 to 11.08 and 14.28 to 4.69 ppm, respectively. The baseline-sensitivity data of these insects to Cry1Ab in laboratory assays was determined. The concentrations of Cry1Ab in the target tissues as studied in seven YieldGard hybrids tested suggest effective management of the two borers. This paper also provides a summary of the expression of the Cry1Ab gene in various genetic backgrounds.