Electrochemical RNA genosensors for toxic algal species: enhancing selectivity and sensitivity

Harmful algal blooms (HABs) are becoming more frequent as climate changes, with tropical species moving northward. Monitoring programs detecting the presence of toxic algae before they bloom are of paramount importance to protect aquatic ecosystems, aquaculture, human health and local economies. Rapid and reliable species identification methods using molecular barcodes coupled to biosensor detection tools have received increasing attention over the past decade as an alternative to the impractical standard microscopic counting-based techniques. This work reports on a PCR amplification-free electrochemical genosensor for the enhanced selective and sensitive detection of RNA from multiple Mediterranean toxic algal species. For a sandwich hybridization (SHA), we designed longer capture and signal probes for more specific target discrimination against a single base-pair mismatch from closely related species and for reproducible signals. We optimized experimental conditions, viz., minimal probe concentration in the SHA on a screen-printed gold electrode and selected the best electrochemical mediator. Probes from 13 Mediterranean dinoflagellate species were tested under optimized conditions and the format further tested for quantification of RNA from environmental samples. We not only enhanced the selectivity and sensitivity of the state-of-the-art toxic algal genosensors but also increased the repertoire of toxic algal biosensors in the Mediterranean, towards an integral and automatic monitoring system.

Fishing gear selectivity for Lates niloticus L., Oreochromis niloticus L. and Rastrineobola argentea P. in Lakes Victoria, Kyoga and Nabugabo

Catch effort data on which fisheries management regulations are sometimes based are not available for most lakes in Uganda. However, failure to regulate fishing gears and methods has been a major cause of collapse of fisheries in the country. Fisheries have been damaged by destructive and non-selective fishing gears and methods such as trawling and beach seining, by use of gill nets of mesh size which crop immature fish and by introduction of mechanised fishing. Selectivity of gears used to crop Lates niloticus L.(Nile perch), Oreochromis niloticus L. (Nile tilapia) and Rastrineobola argentea Pellegrin (Mukene) which are currently the most important commercial species in Uganda were examined in order to recommend the most suitable types, sizes and methods that should be used in exploiting these fisheries. Gill nets of less than 127 mm mainly cropped immature Nile tilapia and Nile perch. To protect these fisheries, the minimum mesh size of gill nets should be set at 127 mm. Seine nets of 5 mm caught high proportions in immature Mukene while those of 10 mm caught mainly mature Mukene. When operated inshore, both sizes caught immature Nile perch and Nile tilapia as by-catch. To protect the Mukene fishery and avoid catching immature bye-catch, a minimum mesh size of the Mukene net should be 10 mm operated as Lampara type net offshore, but since most fishermen have been using 5 mm seine nets for over five years the minimum size should not be allowed to drop below 5 mm pending further thorough investigations. Beach seining and trawling are destructive to fisheries and should be prohibited until data that may justify their use is available.

Is Australia ready for triple gene herbicide stack technology?

Integration of multiple herbicide-resistant genes (trait stacking) into crop plants would allow over the top application of herbicides that are otherwise fatal to crops. The US has just approved Bollgard II® XtendFlex™ cotton which has dicamba, glyphosate and glufosinate resistance traits stacked. The pace of glyphosate resistance evolution is expected to be slowed by this technology. In addition, over the top application of two more herbicides may help to manage hard to kill weeds in cotton such as flax leaf fleabane and milk thistle. However, there are some issues that need to be considered prior to the adoption of this technology. Wherever herbicide tolerant technology is adopted, volunteer crops can emerge as a weed problem, as can herbicide resistant weeds. For cotton, seed movement is the most likely way for resistant traits to move around. Management of multiple stack volunteers may add additional complexity to volunteer management in cotton fields and along roadsides. This paper attempts to evaluate the pros and cons of trait stacking technology by analysing the available literature in other crop growing regions across the world. The efficacy of dicamba and glufosinate on common weeds of the Australian cotton system, herbicide resistance evolution, synergy and antagonisms due to herbicide mixtures, drift hazards and the evolution of herbicide resistance to glyphosate, glufosinate and dicamba were analysed based on the available literature.

Parental exposure to the herbicide diuron results in oxidative DNA damage to germinal cells of the Pacific oyster Crassostrea gigas

Chemical pollution by pesticides has been identified as a possible contributing factor to the massive mortality outbreaks observed in Crassostrea gigas for several years. A previous study demonstrated the vertical transmission of DNA damage by subjecting oyster genitors to the herbicide diuron at environmental concentrations during gametogenesis. This trans-generational effect occurs through damage to genitor-exposed gametes, as measured by the comet-assay. The presence of DNA damage in gametes could be linked to the formation of DNA damage in other germ cells. In order to explore this question, the levels and cell distribution of the oxidized base lesion 8-oxodGuo were studied in the gonads of exposed genitors. High-performance liquid chromatography coupled with UV and electrochemical detection analysis showed an increase in 8-oxodGuo levels in both male and female gonads after exposure to diuron. Immunohistochemistry analysis showed the presence of 8-oxodGuo at all stages of male germ cells, from early to mature stages. Conversely, the oxidized base was only present in early germ cell stages in female gonads. These results indicate that male and female genitors underwent oxidative stress following exposure to diuron, resulting in DNA oxidation in both early germ cells and gametes, such as spermatozoa, which could explain the transmission of diuron-induced DNA damage to offspring. Furthermore, immunostaining of early germ cells seems indicates that damages caused by exposure to diuron on germ line not only affect the current sexual cycle but also could affect future gametogenesis.

Effect of sugarcane cropping systems on herbicide losses in surface runoff

Herbicide runoff from cropping fields has been identified as a threat to the Great Barrier Reef ecosystem. A field investigation was carried out to monitor the changes in runoff water quality resulting from four different sugarcane cropping systems that included different herbicides and contrasting tillage and trash management practices. These include (i) Conventional - Tillage (beds and inter-rows) with residual herbicides used; (ii) Improved - only the beds were tilled (zonal) with reduced residual herbicides used; (iii) Aspirational - minimum tillage (one pass of a single tine ripper before planting) with trash mulch, no residual herbicides and a legume intercrop after cane establishment; and (iv) New Farming System (NFS) - minimum tillage as in Aspirational practice with a grain legume rotation and a combination of residual and knockdown herbicides. Results suggest soil and trash management had a larger effect on the herbicide losses in runoff than the physico-chemical properties of herbicides. Improved practices with 30% lower atrazine application rates than used in conventional systems produced reduced runoff volumes by 40% and atrazine loss by 62%. There were a 2-fold variation in atrazine and >10-fold variation in metribuzin loads in runoff water between reduced tillage systems differing in soil disturbance and surface residue cover from the previous rotation crops, despite the same herbicide application rates. The elevated risk of offsite losses from herbicides was illustrated by the high concentrations of diuron (14mugL-1) recorded in runoff that occurred >2.5months after herbicide application in a 1st ratoon crop. A cropping system employing less persistent non-selective herbicides and an inter-row soybean mulch resulted in no residual herbicide contamination in runoff water, but recorded 12.3% lower yield compared to Conventional practice. These findings reveal a trade-off between achieving good water quality with minimal herbicide contamination and maintaining farm profitability with good weed control.

Hyperspectral sensing to detect the impact of herbicide drift on cotton growth and yield

Yield loss in crops is often associated with plant disease or external factors such as environment, water supply and nutrient availability. Improper agricultural practices can also introduce risks into the equation. Herbicide drift can be a combination of improper practices and environmental conditions which can create a potential yield loss. As traditional assessment of plant damage is often imprecise and time consuming, the ability of remote and proximal sensing techniques to monitor various bio-chemical alterations in the plant may offer a faster, non-destructive and reliable approach to predict yield loss caused by herbicide drift. This paper examines the prediction capabilities of partial least squares regression (PLS-R) models for estimating yield. Models were constructed with hyperspectral data of a cotton crop sprayed with three simulated doses of the phenoxy herbicide 2,4-D at three different growth stages. Fibre quality, photosynthesis, conductance, and two main hormones, indole acetic acid (IAA) and abscisic acid (ABA) were also analysed. Except for fibre quality and ABA, Spearman correlations have shown that these variables were highly affected by the chemical. Four PLS-R models for predicting yield were developed according to four timings of data collection: 2, 7, 14 and 28 days after the exposure (DAE). As indicated by the model performance, the analysis revealed that 7 DAE was the best time for data collection purposes (RMSEP = 2.6 and R2 = 0.88), followed by 28 DAE (RMSEP = 3.2 and R2 = 0.84). In summary, the results of this study show that it is possible to accurately predict yield after a simulated herbicide drift of 2,4-D on a cotton crop, through the analysis of hyperspectral data, thereby providing a reliable, effective and non-destructive alternative based on the internal response of the cotton leaves.

3-Dichloroacetyl oxazolidine protect maize from imazethapyr herbicide injury

Safeners are an important tool used to ensure the safe using of herbicide. The objective of this paper was to investigate the protective effect of four 3-dichloroacetyl oxazolidine safeners (3-dichloroacetyl-2,2-dimethyl-1,3-oxazolidine [R-28725], racemate of 3-dichloroacetyl-2,2-dimethyl-4- ethyl-1,3-oxazolidine, and its two chiral stereoisomers) in reducing the injury caused by imazethapyr. Physiological and biochemical tests were conducted under laboratory condition, by using seed treatment with safeners and soil treatment with imazethapyr, respectively. The interaction of two safeners (R-28725 and R-stereoisomer) and imazethapyr reduced the injury of maize significantly, and also increased glutathione content, activity of glutathione S-transferases, and activity of acetolactate synthase in maize. When induced by R-stereoisomer, the GSH content in root and in shoot increased 100.7% and 73.6%, respectively. When induced by R-28725, the GST activity in vivo increased threefold and the GST activity in vitro more than doubled. The kinetic parameter Vmax of GST in the maize treated with R-28725 and R-stereoisomer increased by 102.2% and 81.9%, respectively, compared with the control. The results also showed that R-28725 and R-stereoisomer induced glutathione S-transferases affinity for the substrate of conjugation reaction significantly.

Assessment of grazing selectivity in relation to land cover: a comparison between sheep and goats

Portugal’s Northeast production of sheep and goats are based on the exploitation of landscape by-products such as spontaneous native vegetation and agriculture leftovers. Shepherds tend the flocks throughout grazing itineraries every day, crossing a mosaic of patches of varied land uses. During the journey, the shepherd acts together with the sheep and goats to select each patch in creating an ordered sequence of land uses. The focus of the research is on the land-use composition of the grazing itineraries; determinate how they depend on the patterns of the landscape mosaic. It is utilized a data set of 26 monthly herd’s itineraries, 13 of sheep and 13 of goats, to investigate the relationship of the land uses crossed by the flocks and the land uses of the landscape, evaluating the land-use preferences and selectivity of the sheep and goats. It is utilized the divergences in the time spent and distance travelled by the herds and the area of the land uses in the landscape, the chi-square test to relate the preferred land used and the season, and the discriminate analysis to distinguish the preferences and the selectivity of the herd of sheep and the herd of goats. The herds of the sheep and the goats presented different land-use preferences over the seasons and the discriminant analysis shows that they have different landscape preferences. The herd of sheep has the highest selectivity indexes for the annual irrigated crops, the agricultural complex systems and the agroforestry land uses. The highest selectivity indexes for the herd of goats were found for the deciduous forest, the agriculture with natural and semi-natural spaces and the shrublands land uses. It was concluded that the landscape management for sheep and goats herding has to be different: the agricultural land uses are essential to the flocks of sheep and the forest land uses are decisive to the flocks of goats.

Dual layer solid phase extraction based on molecular imprinting technology: Seeking a route to enhance selectivity for trace analysis of pesticide residues in olive oil.

Aiming to introduce a multiresidue analysis for the trace detection of pesticide residues belonging to organophosphorus and triazine classes from olive oil samples, a new sample preparation methodology comprising the use of a dual layer of “tailor-made” molecularly imprinted polymers (MIPs) SPE for the simultaneous extraction of both pesticides in a single procedure has been attempted. This work has focused on the implementation of a dual MIP-layer SPE procedure (DL-MISPE) encompassing the use of two MIP layers as specific sorbents. In order to achieve higher recovery rates, the amount of MIP layers has been optimized as well as the influence of MIP packaging order. The optimized DL-MISPE approach has been used in the preconcentration of spiked organic olive oil samples with concentrations of dimethoate and terbuthylazine similar to the maximum residue limits and further quantification by HPLC. High recovery rates for dimethoate (95%) and terbuthylazine (94%) have been achieved with good accuracy and precision. Overall, this work constitutes the first attempt on the development of a dual pesticide residue methodology for the trace analysis of pesticide residues based on molecular imprinting technology. Thus, DL-MISPE constitutes a reliable, robust, and sensitive sample preparation methodology that enables preconcentration of the target pesticides in complex olive oil samples, even at levels similar to the maximum residue limits enforced by the legislation.

Trammel net selectivity studies in the Algarve (southern Portugal), gulf of cadiz (Spain), Basque coutry (Spain) and cyclades islands (Greece)

Trammel net fisheries were studied in four areas: the Cantabrian Sea (Basque Country, Spain), the Algarve (Southern Portugal), the Gulf of Cádiz (Spain) and the Cyclades Islands (Greece). Surveys were carried out in order to identify trammel net métiers and to characterise the gear used. Trammel nets were among the most important gears used in the small-scale fisheries, with up to 9 different métiers identified in each area. The most important métiers in the Algarve and the Gulf of Cádiz were those for cuttlefish (Sepia officinalis) and soles (Solea senegalensis, Microchirus azevia, Synaptura lusitanica). In the Cantabrian Sea, sole (Solea vulgaris), shellfish (several species) and scorpion fish (Scorpaena spp.) métiers dominated while a variety of species where targeted in the multi-species trammel net fishery in the Cyclades. In each area, experimental trammel nets of six different types (combinations of 2 large mesh outer panel sizes and 3 small mesh outer panels) corresponding to the most common métier, were constructed and fishing trials carried out on a seasonal basis (4 seasons in the Cantabrian Sea, Algarve and Cyclades and 2 in the Gulf of Cádiz) using chartered commercial fishing vessels. Overall, 271, 360, 185 and 185 km of trammel nets were fished in the experimental fishing trials in the Cantabrian Sea, Algarve, Gulf of Cádiz and Cyclades Islands respectively.

Selectivity and efficiency of herbicides in weed control on sweet sorghum.

A carência de herbicidas registrados para o controle de plantas daninhas, em sorgo sacarino, torna-se fator limitante à produção. Objetivou-se avaliar a seletividade de herbicidas em sorgo sacarino e a eficiência no controle de plantas daninhas. O delineamento experimental foi de blocos ao acaso, com quarto repetições. Os tratamentos incluíram cultivares de sorgo sacarino (BRS 506, BRS 509 e BRS 511) e herbicidas aplicados isoladamente em pré-emergência (atrazine - 1.500 g ha-1; atrazine + s-metolachlor - 1.665 g ha-1 + 1.035 g ha-1; flumioxazin - 50 g ha-1; s-metolachlor - 1.440 g ha-1) e pós-emergência (tembotrione - 100,8 g ha-1) e de forma sequencial em pré e pós-emergência (atrazine + s-metolachlor + tembotrione - 1.665 g ha-1 + 1.035 g ha-1 + 100,8 g ha-1; atrazine + tembotrione - 1.500 g ha-1 + 100,8 g ha-1; flumioxazin + tembotrione - 50 g ha-1 + 100,8 g ha-1), mais duas testemunhas, uma capinada e outra sem capina. A fitotoxicidade ao sorgo foi de 98,0 %, 98,0 % e 100 % de tembotrione; 100 %, 98,7 % e 100 % de flumioxazin + tembotrione; 100 %, 100 % e 100 % de atrazine + s-metolachlor + tembotrione; e 98,7 %, 98,7 % e 99,7 % de atrazine + tembotrione, respectivamente para as cultivares BRS 509, BRS 506 e BRS 511. Tembotrione, flumioxazin + tembotrione, atrazine + s-metolachlor + tembotrione e atrazine + tembotrione apresentaram bom controle de Ipomoea indivisa, Digitaria ciliaris e Urochloa plantaginea, porém, não demonstraram seletividade às cultivares de sorgo. O atrazine, de modo geral, apresentou maior seletividade aos componentes de rendimento das cultivares BRS 509 e BRS 506.

Chronic exposure to sub-lethal concentration of a glyphosate-based herbicide alters hormone profiles and affects reproduction of female Jundia (Rhamdia quelen)

This work was carried out to verify the effect of a glyphosate-based herbicide on Jundia hormones (cortisol, 17 beta-estradiol and testosterone), oocyte and swim-up fry production. Earthen ponds containing Jundia females were contaminated with glyphosate (3.6 mg/L); blood samples were collected from eight females from each treatment immediately before, or at 1, 10, 20 30 and 40 days following contamination. A typical post-stress rise in cortisol levels was observed at the 20th and 40th days following exposure to glyphosate. At the 40th day, 17 beta-estradiol was decreased in the exposed females. A similar number of oocytes were stripped out from females from both groups, however, a lower number of viable swim-up fry were obtained from the herbicide exposed females, which also had a higher liver-somatic index (LSI). The results indicate that the presence of glyphosate in water was deleterious to Rhamdia quelen reproduction, altering steroid profiles and egg viability. (c) 2006 Elsevier B.V. All rights reserved.

Trends in vertical movement of hexazinone herbicide considering typic eutrorthox soil of Espraiado´s stream cultivated with sugar-cane crop - local and default data scenarios.

2009

PROLINE-BASED CHIRAL STATIONARY PHASES: INSIGHTS INTO THE MECHANISM OF CHIRAL SELECTIVITY

Proline (Pro) is a unique amino acid that has been examined previously as a potential chiral selector for high-performance liquid chromatography. In recent years, a new class of promising Pro based enantioselective stationary phases has been studied and the longer peptides were found to be competitive with commercial chiral stationary phases (CSPs). Here, we aim to perform a comprehensive examination of a t-butoxycarbonyl- (t-Boc-) terminated monoproline selector. This selector was grafted through an amide linkage to an aminopropyl siloxane-terminated Si (111) wafer and to a silicon atomic force microscopy tip. To ensure a flat, homogeneous overlayer of selectors suitable for force spectrometric measurements, the prepared surfaces were characterized using XPS, AFM and contact angle measurements. Chemical force spectrometry (CFS) has been used to examine the chiral discrimination in our monoproline CSP by measuring the interaction forces between two D- or L-monoproline monolayers in water and in the presence of a series of amino acids in solution to explore the degree to which binding of amino acids impacts self-selectivity. Chemical force titration (CFT) has been used to observe the influence of variations in pH on the binding interaction of proline modified chiral surfaces. Here we aim to explore the connection between side-chain hydrophobicity and differences in the nature of the binding between different ionic forms of amino acids and the t-Boc-Pro interface, and thereby to gain insight into the mechanism of chiral selectivity. The CFS results show several trends for different proline selector/amino acid combinations and indicate that the binding characteristics of amino acid to the proline surface is strongly dependent on the amino acid side chain where hydrophilic side chain amino acids exhibit a selectivity opposite to that seen for those with hydrophobic side chains. The CFT studies also provide valuable insights into interactions between the proline selector and the amino acids under a wide range of pH conditions, indicating that protonated amine groups of alanine and serine are closely involved in the binding mechanism to proline surfaces. On the other hand, the presence of the second carboxylic group in aspartic acid plays an important role while interacting with proline.