Effect of Oil Overlay on Inhibition Potential of Roscovitine in Sheep Cumulus‐Oocyte Complexes

Inhibitors of cyclin‐dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus‐oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 μm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor‐free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (?) at 38.5°C and 5% CO₂. At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/?) exhibited total expansion while in both Rosco groups (+/?) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/?). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.     

Multiresidue analysis of seven anticoagulant rodenticides by high-performance liquid chromatography/electrospray/mass spectrometry

Mice and rat populations are commonly controlled by two classes of rodenticide anticoagulants, coumarins and indandiones. However, poisoning of nontarget animals also often occurs. For cases such as these, a rapid, multiresidue method, which provides positive confirmation for both classes of anticoagulant rodenticides, is needed by diagnostic laboratories. A method was developed for the determination of seven anticoagulant rodenticides, coumafuryl, pindone, warfarin, diphacinone, chlorophacinone, bromadiolone, and brodifacoum, in diverse matrices, animal feed, cooked beef, and fruit-flavored beverages using high-performance liquid chromatography/electrospray/mass spectrometry. Detection was by MS/MS with electrospray ionization in negative mode. Confirmation was by retention time, m/z of molecular ion, and two parent-daughter transitions. Recoveries from selected the matrices ranged from 61 to 117%. Limits of quantitation were as low as 1.5-4.5 ng g-1. The developed method was rapid and provided the simultaneous confirmation and quantification of the seven anticoagulant rodenticides.     

Fate of diuron and terbuthylazine in soils amended with two-phase olive oil mill waste

The addition of organic amendments to soil increases soil organic matter content and stimulates soil microbial activity. Thus, processes affecting herbicide fate in the soil should be affected. The objective of this work was to investigate the effect of olive oil production industry organic waste (alperujo) on soil sorption-desorption, degradation, and leaching of diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] and terbuthylazine [N2-tert-butyl-6-chloro-N4-ethyl-1,3,5-triazine-2,4-diamine], two herbicides widely used in olive crops. The soils used in this study were a sandy soil and a silty clay soil from two different olive groves. The sandy soil was amended in the laboratory with fresh (uncomposted) alperujo at the rate of 10% w/w, and the silty clay soil was amended in the field with fresh alperujo at the rate of 256 kg per tree during 4 years and in the laboratory with fresh or composted alperujo. Sorption of both herbicides increased in laboratory-amended soils as compared to unamended or field-amended soils, and this process was less reversible in laboratory-amended soils, except for diuron in amended sandy soil. Addition of alperujo to soils increased half-lives of the herbicides in most of the soils. Diuron and terbuthylazine leached through unamended sandy soil, but no herbicide was detected in laboratory-amended soil. Diuron did not leach through amended or unamended silty clay soil, whereas small amounts of terbuthylazine were detected in leachates from unamended soil. Despite their higher sorption capacity, greater amounts of terbuthylazine were found in the leachates from amended silty clay soils. The amounts of dissolved organic matter from alperujo and the degree of humification can affect sorption, degradation, and leaching of these two classes of herbicides in soils. It appears that adding alperujo to soil would not have adverse impacts on the behavior of herbicides in olive production.     

Metsulfuron methyl sorption-desorption in field-moist soils

Pesticide sorption coefficients (K(d)) are generally obtained using batch slurry methods. As a consequence, the results may not adequately reflect sorption processes in field-moist or unsaturated soil. The objective of this study was to determine sorption of metsulfuron methyl, a weak acid, in field-moist soils. Experiments were performed using low density (i.e., 0.3 g mL(-)(1)) supercritical fluid carbon dioxide (SF-CO(2)) to convert anionic metsulfuron methyl to the molecular species and remove it from the soil water phase only, thus allowing calculation of sorption coefficients (K(d)) at low water contents. K(d) values for sorption of the metsulfuron methyl molecular species on sandy loam, silt loam, and clay loam soil at 11% water content were 120, 180, and 320 mL g(-)(1), respectively. Using neutral species K(d) values, the pK(a) of metsulfuron methyl, and the pH of the soil, we could successfully predict the K(d) values obtained using the batch slurry technique, which typically has a predominance of anionic species in solution during the sorption characterization. This application of supercritical fluid extraction to determine sorption coefficients, combined with sulfonylureas' pK(a) values and the soil pH, will provide an easy method to predict sorption in soil at different pH levels.     

Adsorption of pesticides from water by functionalized organobentonites

Replacement of natural inorganic cations of clay minerals with organic cations has been proposed as a strategy to improve the adsorptive capacity of clay minerals for organic compounds, including pesticides. The organic cations most commonly used for this purpose have been quaternary ammonium ions containing alkyl or aryl chains without specific functional groups. In this work, we evaluated the ability of two bentonites (SWy-2 and SAz-1) exchanged with four natural organic cations containing diverse functional groups (L-carnitine, L-cysteine ethyl ester, L-cystine dimethyl ester, and thiamine) as adsorbents of pesticides varying in their chemical structures (simazine, hexazinone, triadimefon, alachlor, carbaryl, and imazethapyr). For comparison purposes, the adsorptive properties of two "classical" organobentonites, hexadecyltrimethylammonium- and phenyltrimethylammonium-exchanged bentonites, were also determined. Most organobentonites displayed higher affinity for the pesticides than the untreated bentonites, but the improvement in adsorption capacity varied depending on the characteristics of the pesticide and the interlayer organic cation. Triadimefon, carbaryl, and imazethapyr displayed the highest affinity for carnitine (K(f) = 229-2377)-, thiamine (K(f) = 83-354)-, and cystine (K(f) = 96-100)-treated bentonites, respectively, whereas alachlor was adsorbed similarly by all organobentonites. In general, pesticide adsorption-desorption hysteresis was greater for adsorbents with the highest adsorption capacities. The results demonstrate that selective modification of smectitic clay minerals with natural organic cations containing appropriate functional groups can be a useful strategy to improve their performance for the removal of specific pesticides from the environment.     

Follow-up of mercury levels in fish,human hair and urine in the Madeira and Tapajós basins,Amazon, Brazil

Informal economy of gold mining has contaminated some important river basins in Amazon. Follow-up studies on critical compartments showed some areas with high Hg levels in fish as well as in human hair samples. Average Hg in piscivorous fish in the Madeira river itself was 846 ppb (N=284) with a maximum of 3921 ppb. Mercury in fish from non polluted areas in this basin shows high variability, even for single species. A seasonal variation in Hg content was observed, with higher values at the end of the dry season. In the upper Tapajós basin comparable values were found for fish but with a definite decreasing trend downstream. Average value for piscivorous fish in the whole Tapajós basin is 482 ppb (N=122) with a maximum value of 3770 ppb. Hair Hg was higher in fishing villages in the Tapajós (average: 17 ppm; with N=432 and maximum value of 176 ppm) than in the Madeira (average: 9 ppm; N=169; maximum 71 ppm), and data from some areas of the Tapajós suggest a decrease with time. Mercury was much higher in urine of goldshop workers in Santarém (low Tapajós) than in Alta Floresta (high Tapajbs) and show a decreasing trend in both cases, probably related to the significant decline in gold mining activities during the study period (1986–1994).     

Bioavailability of organoclay formulations of atrazine in soil

Pesticide formulations based on organoclays have been proposed to prolong the efficacy and reduce the environmental impact of pesticides in soil. This research addressed the question of whether atrazine in organoclay-based formulations is irreversibly sorbed or is bioavailable for bacterial degradation in soil. Different cations of l-carnitine (CAR), tyramine (TYRAM), hexadimethrine (HEXADIM), phenyltrimethylammonium (PTMA), hexadecyltrimethylammonium (HDTMA), and Fe(III) were incorporated into Na-rich Wyoming montmorillonite (SWy-2) and Ca-rich Arizona montmorillonite (SAz-1) at 100% of the cation exchange capacity (CEC) of the clays as a strategy to enhance the affinity of the clay minerals for atrazine. A Buse loam soil from Becker, MN, was treated with three organoclay-based formulations of 14C-atrazine or free herbicide and incubated for 2 weeks. To determine the bioavailability of 14C-atrazine, the soil was inoculated with Pseudomonas sp. strain ADP, which rapidly mineralizes atrazine. At day 0, and after a 2 week incubation, mineralization and the amount of 14C-atrazine residues distributed between the aqueous-extractable, methanol-extractable, and bound fractions in the soil were determined to characterize the availability of nonaged and aged atrazine residues. By the end of the 2 week incubation, the microorganisms had mineralized >80% of the initial readily available (water-extractable) and >70% of the less readily available (methanol-extractable) 14C-atrazine in the soil. Bound residues increased from <4% at day 0 to ～17% after the 2 week incubation for both the formulated and free forms of atrazine. The results of these incubation experiments show that the bioavailabilities of atrazine were similar in the case of the organoclay formulations and as free atrazine. This indicated that whereas more atrazine was sorbed and less likely to be transported in soil, when formulated as organoclay complexes, it was ultimately accessible to degrading bacteria, so that the herbicide is likely to be naturally attenuated by soil microorganisms.     

Influence of soil pH-sorption interactions on imazethapyr carry-over

Soil pH affects imazethapyr sorption-desorption, which in turn can affect persistence and bioavailability. Long-term imazethapyr carry-over has been observed in soil that is below pH 6.5, resulting in significant sugarbeet damage. Imazethapyr concentration decreased rapidly in field soil, regardless of pH. Despite similar amounts of imazethapyr remaining in aged soils at different pH levels, there were differences in bioavailability, which can be explained by sorption-desorption. At low pH more imazethapyr was sorbed than at high pH, but it readily desorbed. At high pH less imazethapyr was sorbed initially, but it did not readily desorb. Thus, after 3 months, the remaining imazethapyr in low-pH soil was desorbable and bioavailable, resulting in injury to canola and sugarbeet. Liming aged, low-pH soil released bound imazethapyr residues, which would then be degraded and result in less carry-over.     

Processes affecting the dissipation of the herbicide isoxaflutole and its diketonitrile metabolite in agricultural soils under field conditions

Two-year field dissipation studies were conducted in three soil types in Minnesota to examine the processes affecting the dissipation of the herbicide isoxaflutole and its phytotoxic diketonitrile metabolite (DKN) under relatively cool, wet soil conditions. Plots of cuphea were treated with isoxaflutole and potassium bromide, a nonsorbed, nondegraded tracer. Replicate soil cores were collected six times during the growing season to a depth of 1 m, and the bromide or herbicide concentration was measured in each of five depth increments. The dissipation half-life (DT50) of isoxaflutole + DKN was 8-18 days in each soil. Bromide and herbicide concentrations were low at depths >40 cm throughout the study, and herbicide concentrations in soil 100 days after application were usually undetectable. Simulation modeling using Hydrus-1D for the loam soil suggested that plant uptake was an important mechanism of dissipation.     

Onset of luteal activity in foaling and seasonally anoestrous mares treated with artificial light.

Sixty-four mares (27 foaling, 37 barren or maiden), mainly Finnhorses, were subjected to treatment with 14.5 h of light and 9.5 h of darkness, starting at the beginning of December. The onset of cycling in non-foaling mares was estimated by weekly serum progesterone determinations. All of the non-foaling mares cycled in the middle of March. They started to cycle on average in the middle of February, 11.1 weeks after the beginning of the trial. There were statistically significant differences in relation to breed (Finnhorses started to cycle 2 weeks later than warm blooded, p less than 0.02) and in relation to age (brood mares with mean age 10 years, started to cycle 10 days later than those 3 years old, p less than 0.03). However, there was no statistical significant relation to previous lactation, although lactating mares lactating up to the previous autumn started to cycle 7 days later than dry mares (p less than 0.15). Artificial insemination of 14 mares in the 2nd oestrus of the year, in March, resulted in 12 foalings in the subsequent year (86%). The following winter, all pregnant mares (N = 27) were exposed to the same kind of light treatment, starting on 1st December. The 1st mare foaled at the end of January. The time from foaling to 1st post partum ovulation was significantly longer (17.0 days) in foalings taking place within 10 weeks from the beginning of the light treatment period, than in foalings occurring after more than 10 weeks of lighting (12.1 days) (p less than 0.01).