Effects of thiobencarb herbicide to an alga (Nannochloris oculata) and the cladoceran (Daphnia magna)

Chronic toxicity studies were conducted with an algae (Nannochloris oculata) and the cladoceran (Daphnia magna) to determine their relative sensitivities to the thiocarbamate herbicide thiobencarb (S-4-chlorobenzyl diethylthiocarbamate). Most of the algal populations were initially affected by exposure to the herbicide. Thiobencarb concentrations higher than 0.5 mg/L significantly reduced algal densities after 24-h exposure. The 24-h static EC50 in D. magna was 3.01 mg/L. The sublethal effects of 0.3, 0.37, 0.5, 0.75, and 1.5 mg/L of thiobencarb concentrations on the survival, reproduction, and growth of D. magna were monitored for 21 days. The parameters used to determined the effect of the herbicide on D. magna were mean total young per female; mean brood size; days to first brood; intrinsic rate of natural increase (r); growth; and survival. Reproduction was significantly reduced at thiobencarb concentrations of 0.30 mg/L and higher while survival was affected after exposure to 0.75 and 1.5 mg/L of the pesticide. The r value decreased with increasing concentrations of thiobencarb. Growth, as measured by body length, was depressed significantly after exposure to all herbicide concentrations tested.     

Effects of low concentrations of the phenylurea herbicide diuron on biofilm algae and bacteria

A system of recirculating channels was used in this study to examine the long-term effects (29 d) of environmentally realistic concentrations of the herbicide diuron (from 0.07 to 7 μg L⁻¹) on biofilm communities. The autotrophic activity of biofilms was affected by this herbicide, as reflected by a marked decrease in the photosynthetic efficiency. Diuron exposure also increased chlorophyll-a content and reduced the biovolume of diatom taxa at low concentrations. The effects on bacteria were also remarkable. Bacterial abundance was reduced after a week of exposure to the herbicide at a range of concentrations. Effects were on the number of live bacteria and on the increase in the leucine-aminopeptidase activity. It is suggested that inputs of herbicides to the river ecosystem at low concentrations may cause a chain of effects in the biofilm, which include inhibitory effects on algae but also indirect effects on the relationships between biofilm components.     

Pollution-induced community tolerance (PICT) and seasonal variations in the sensitivity of phytoplankton to atrazine in nanocosms.

Algae communities exposed to a herbicide like atrazine (PS II inhibitor) are expected to be selected and to be more tolerant to the herbicide than unexposed communities (pollution-induced community tolerance, PICT). The PICT may be an ecotoxicological tool for detecting this selective action of chronic pollution, and this method has been applied to several toxicants in experimental systems and in field studies. But the detection of PICT with PS II inhibitors has sometimes been variable. This work was done to study the long-term effects of exposure to atrazine (10 microg/l) and the PICT responses of phytoplankton communities in repeated outdoor nanocosms. Phytoplankton communities were sampled in Lake Geneva at different periods of the year and the effects of atrazine were analysed by studying community structure, biomass and primary production, and by measuring tolerance to atrazine in a short-term physiological test based on 14C incorporation. We find that PICT is a sensitive method for measuring effects. Even atrazine concentrations causing little restructuring induced tolerance in most of our experiments. But the short- and long-term responses of phytoplankton to atrazine varied between experiments, probably due to the initial compositions of the communities and environmental factors associated with seasonal parameters. The selection and detection steps of PICT to atrazine thus vary greatly with environmental conditions and the physiological adaptations of algae to the herbicide. To monitor risk assessment in aquatic systems, PICT studies applied to algae, must be investigated in the light of seasonal contaminations and seasonal events and successions.     

The herbicide paraquat induces alterations in the elemental and biochemical composition of non-target microalgal species

Huge quantities of pesticides are dispersed in the environment, affecting non-target organisms. Since paraquat affects the photosynthetic process, the biochemical composition of affected species should be altered. The effect of paraquat on Chlamydomonas moewusii, a freshwater non-target species, was studied. After 48 h of herbicide exposure, growth rate, dry weight, and chlorophyll a and protein content were affected by paraquat concentrations above 0.05 μM. C/N ratio was also affected due to a decrease in nitrogen content in the dry biomass, while the carbon content remained constant for all paraquat concentrations assayed. Enzymes involved in nitrogen assimilation were affected by paraquat, being nitrate reductase activity more sensitive to paraquat than nitrite reductase. Based on the results obtained in the present study, paraquat exerts adverse effects upon a common freshwater green microalga, thus the application of this herbicide for weed control must be carried out very carefully, so that any disturbance affecting algae will have severe repercussions on higher trophic levels and on the elemental biogeochemical cycles.     

Sucrose amendment enhances phytoaccumulation of the herbicide atrazine in Arabidopsis thaliana

Growth in the presence of sucrose was shown to confer to Arabidopsis thaliana (thale cress or mustard weed) seedlings, under conditions of in vitro culture, a high level of tolerance to the herbicide atrazine and to other photosynthesis inhibitors. This tolerance was associated with root-to-shoot transfer and accumulation of atrazine in shoots, which resulted in significant decrease of herbicide levels in the growth medium. In soil microcosms, application of exogenous sucrose was found to confer tolerance and capacity to accumulate atrazine in Arabidopsis thaliana plants grown on atrazine-contaminated soil, and resulted in enhanced decontamination of the soil. Application of sucrose to plants grown on herbicide-polluted soil, which increases plant tolerance and xenobiotic absorption, thus appears to be potentially useful for phytoremediation.     

Effects of a sulfonylurea herbicide on the soil bacterial community

Sulfonylurea herbicides are widely used on a wide range of crops to control weeds. Chevalier® OnePass herbicide is a sulfonylurea herbicide intensively used on cereal crops in Algeria. No information is yet available about the biodegradation of this herbicide or about its effect on the bacterial community of the soil. In this study, we collected an untreated soil sample, and another sample was collected 1 month after treatment with the herbicide. Using a high-resolution melting DNA technique, we have shown that treatment with Chevalier® OnePass herbicide only slightly changed the composition of the whole bacterial community. Two hundred fifty-nine macroscopically different clones were isolated from the untreated and treated soil under both aerobic and microaerobic conditions. The strains were identified by sequencing a conserved fragment of the 16S rRNA gene. The phylogenetic trees constructed using the sequencing results confirmed that the bacterial populations were similar in the two soil samples. Species belonging to the Lysinibacillus, Bacillus, Pseudomonas, and Paenibacillus genera were the most abundant species found. Surprisingly, we found that among ten strains isolated from the treated soil, only six were resistant to the herbicide. Furthermore, bacterial overlay experiments showed that only one resistant strain (related to Stenotrophomonas maltophilia) allowed all the sensitive strains tested to grow in the presence of the herbicide. The other resistant strains allowed only certain sensitive strains to grow. On the basis of these results, we propose that there must be several biodegradation pathways for this sulfonylurea herbicide.     

Biodegradation of the herbicide glyphosate by filamentous fungi in platform shaker and batch bioreactor

The biodegradation conducted by microorganisms on herbicide glyphosate (N-phosphonomethylglycine) was investigated. Five strains of filamentous fungi belonging to the Fusarium genre were grown on Czapeck medium without phosphorous and supplemented with the addition of glyphosate. The assays were conducted to determine the ability of use as a phosphorous source, the inhibition caused by presence of herbicide, and the biodegradation in shaker and bioreactor by Fusarium strains. It was observed that the herbicide did not show any negative effect on microrganisms by quantity of the biomass. Among the strains tested, no inhibition was noted by the addition of glyphosate even at a high concentration. All strains studied were able to biodegrade it and use the herbicide as a phosphorous source. The formation of consortium was not better than the strains tested in pure culture. The biodegradation in the bioreactor was better than in the shaker. However, there wasn't any influence on biodegradation rate by changing the amount of oxygen in the system.     

Biodegradation of the herbicide glyphosate by filamentous fungi in platform shaker and batch bioreactor

The biodegradation conducted by microorganisms on herbicide glyphosate (N-phosphonomethylglycine) was investigated. Five strains of filamentous fungi belonging to the Fusarium genre were grown on Czapeck medium without phosphorous and supplemented with the addition of glyphosate. The assays were conducted to determine the ability of use as a phosphorous source, the inhibition caused by presence of herbicide, and the biodegradation in shaker and bioreactor by Fusarium strains. It was observed that the herbicide did not show any negative effect on microrganisms by quantity of the biomass. Among the strains tested, no inhibition was noted by the addition of glyphosate even at a high concentration. All strains studied were able to biodegrade it and use the herbicide as a phosphorous source. The formation of consortium was not better than the strains tested in pure culture. The biodegradation in the bioreactor was better than in the shaker. However, there wasn't any influence on biodegradation rate by changing the amount of oxygen in the system.     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N2-ethyl-N?-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79-82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53-83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Biodegradation of s-triazine herbicide atrazine by Enterobacter cloacae and Burkholderia cepacia sp. from long-term treated sugarcane-cultivated soils in Kenya

In this study soils from sugarcane-cultivated fields were screened for bacterial species capable of atrazine (6-chloro-N²-ethyl-N⁴-isopropyl-1,3,5-triazine-2,4-diamine) degradation due to long exposure of the soils to this herbicide. To enrich for atrazine degraders, Minimal Salt Medium containing atrazine as the sole N source and glucose as the C source was inoculated with soils impacted with this herbicide and incubated. Bacterial growth was monitored by measuring optical density. The degradation of atrazine was followed by measuring residual atrazine in liquid cultures over a given time period by high performance liquid chromatography. Bacterial strains isolated from the enrichment cultures were characterized by biochemical tests and identified by 16S rRNA gene sequencing. Two bacterial strains coded ISL 8 and ISL 15 isolated from two different fields were shown to have 94 and 96% 16S rRNA gene sequence similarity to Burkholderia cepacia respectively. Another bacterial sp., ISL 14 was closely related to Enterobacter cloacae with a 96% 16S rRNA gene sequence similarity. There was not much difference between the extents of atrazine degradation by the enrichment cultures with communities (79–82% applied amount) from which pure strains were isolated and the pure strains themselves in liquid cultures that showed a degradation of 53–83% of applied amount. The study showed existence of bacterial strains in different sugarcane-cultivated fields which can use atrazine as a nitrogen source. The bacterial strains isolated can be used to enhance the degradation of atrazine in contaminated soils where atrazine is still considered to be recalcitrant.     

Variations of bacterial community structure in flooded paddy soil contaminated with herbicide quinclorac

The denaturing gradient gel electrophoresis (DGGE) method was applied to determine the relative genetic complexity of microbial communities in flooded paddy soil treated with herbicide quinclorac (3,7-dichloro-8-quinoline-carboylic acid). The results obtained showed a significant effect of quinclorac on the development of bacterial populations in soils contaminated with different concentrations of the herbicide at the early time after application. In general, however, the number of populations of the same soil sample treated with the same concentration of the quinclorac differed obviously with increasing incubation time within the early 8 weeks. The scale of differences in banding patterns-showed that the microbial community structures of the quinclorac-treated and non-quinclorac-treated soils were not significantly different after 21 weeks of incubation. Quantification, as demonstrated in this paper, was studied by establishing dose-response relationships. Significant pattern variations were quantified. Prominent DGGE bands were excised, cloned and sequenced to gain insight into the identities of predominant bacterial populations. The majority of DGGE band sequences were related to bacterial genera Clostridium, Sphingobacterium, Xanthomonas and Rhodococcus.     

A microcosm system to evaluate the toxicity of the triazine herbicide simazine on aquatic macrophytes

This study evaluates the effects of the triazine herbicide simazine in an outdoor pond microcosm test system that contained two submerged rooted species (Myriophyllum spicatum and Elodea canadensis) and two emergent rooted species (Persicaria amphibia and Glyceria maxima) over a period of 84 days. Simazine was applied to the microcosms at nominal concentrations of 0.05, 0.5 and 5 mg/L. General biological endpoints and physiological endpoints were used to evaluate herbicide toxicity on macrophytes and the algae developing naturally in the system.Concentration-related responses of macrophytes and algae were obtained for the endpoints selected, resulting in a no observed ecologically adverse effect concentration (NOEAEC) at simazine concentrations of 0.05 mg active ingredient/L after 84 days. E. canadensis was the most negatively affected species based on length increase, which was consistently a very sensitive parameter for all macrophytes. The experimental design presented might constitute a suitable alternative to conventional laboratory single-species testing.     

Engineering the metabolism of the phenylurea herbicide chlortoluron in genetically modified Arabidopsis thaliana plants expressing the mammalian cytochrome P450 enzyme CYP1A2

Transgenic Arabidopsis thaliana plants were generated by introduction of the human P450 CYP1A2 gene, which metabolizes a number of herbicides, insecticides and industrial chemicals. Transgenic A. thaliana plants expressing CYP1A2 gene showed remarkable resistance to the phenylurea herbicide chlortoluron (CTU) supplemented either in plant growth medium or sprayed on foliar parts of the plants. HPLC analyses showed a strong reduction in CTU accumulation in planta supporting the tolerance of transgenic lines to high concentrations of CTU. Besides increased herbicide tolerance, expression of CYP1A2 resulted in no other visible phenotype in transgenic plants. Our data indicate that CYP1A2 can be used as a selectable marker for plant transformation, allowing efficient selection of transgenic lines in growth medium and/or in soil-grown plants. Moreover, these transgenic plants appear to be useful for herbicide resistance as well as phytoremediation of environmental contaminants.     

Chitosan/glyphosate formulation as a potential,environmental friendly herbicide with prolonged activity

Although there are many reports on the dangers posed by glyphosate, this herbicide is still one of the most popular and widely used total weed killers. Therefore, great effort should be made to minimize exposure to this herbicide and limit its losses into the environment. The aim of this study was to prepare a new formulation consisted of various molecular weight chitosans with glyphosate and their evaluation toward active substance release, phytotoxicity, and preliminary herbicidal efficiency. The phytotoxicity study of the obtained chitosan/glyphosate formulations was determined based on the Organisation for Economic Co-operation and Development 208 guideline. Among all tested formulations evaluated for phytotoxicity, that containing the highest molecular weight of chitosan was found to be the least toxic, showing simultaneously the most effective herbicidal activity against selected weeds. The release rate of glyphosate from the obtained formulations was dependent on the molecular weight and viscosity of chitosan.     

Growth rate effects, responses of antioxidant enzymes and metabolic fate of the herbicide Propanil in the aquatic plant Lemna minor

Propanil (3,4-dichloropropionanilide) is a selective contact pesticide, recommended for post-emergence use in rice. This herbicide may end up in surface waters and present potential risk for aquatic vascular plants. Therefore, its toxicity was evaluated on Lemna minor L., an aquatic plant regularly used for toxicological studies, during time- and concentration-dependent exposure. Toxicity assessments were based on inhibition of growth of L. minor cultures after 24 days. The obtained results showed that the growth of Lemna was affected by the herbicide. The responses of the guaiacol peroxidase (G-POD) and glutathione S-transferase (GST) involved in the xenobiotic metabolism and antioxidative system were also investigated following Propanil exposure. Our results showed that Propanil has not induced enzymatic antioxidative defenses of L. minor. Both 3,4-dichloroaniline (3,4-DCA) and 3,4-dichloroacetanilide are the major metabolites in this plant. On the contrary, only 3,4-DCA was found in culture media after 4 days. Probably, the enzymatic hydrolysis by acyl acylamidase and the acetylation by acetyl-CoA are the major pathways for these transformation products, respectively. The results of this study showed that the selected aquatic plant has the potential to accumulate and metabolize rice herbicide, like Propanil. Based on these toxicity data this herbicide should impair the establishment of non-target aquatic plants.     

Predator cues and an herbicide affect activity and emigration in an agrobiont wolf spider

Animals use chemical cues for signaling between species. However, anthropogenic chemicals might interrupt this natural chemical information flow, with potential impacts on predator-prey interactions. Our goal was to explore how Buccaneer® Plus, a common herbicide similar to Round-up® (active ingredient glyphosate), affected the interactions between intraguild predators. The wolf spider Pardosa milvina (Hentz, 1844) is numerically dominant in agricultural systems across the eastern United States, and often falls prey to or competes with the larger wolf spider, Hogna helluo (Walckenaer, 1837) and/or the carabid beetle, Scarites quadriceps (Chaudoir, 1843). We tested the effects of chemical cues from these intraguild predators and exposure to herbicide on the activity, emigration, and survival of P. milvina using a full-factorial laboratory experiment. Both predator cues and herbicide led to a decrease in movement by P. milvina. However, although H. helluo cues alone decreased movement, S. quadriceps cues only decreased movement when combined with herbicide. These results indicate that predation risk and herbicide application likely interact in complex ways to affect the movement of a major arthropod predator in agricultural systems, and thus may have complex effects on the food web.     

On the way to cyanobacterial blooms: impact of the herbicide metribuzin on the competition between a green alga (Scenedesmus) and a cyanobacterium (Microcystis)

The hypothesis that exposure to a common and widely applied photosynthesis-inhibiting herbicide, metribuzin, would alter the outcome of the competitive battle between susceptible green algae (Scenedesmus obliquus) and tolerant cyanobacteria (Microcystis aeruginosa) was tested. In a long-term (17 d) experiment, Scenedesmus and Microcystis populations as well as mixtures that started with different inoculum composition (i.e. 3:1, 1:1 and 1:3 Scenedesmus:Microcystis) were grown in the absence or presence of metribuzin (100 microg l-1). In the absence of metribuzin, Scenedesmus was competitively superior and out-competed Microcystis regardless the initial composition of the mixed communities. However, this competitive outcome was reversed completely in the presence of metribuzin, where despite growth inhibition Microcystis became dominant. Hence, photosynthesis-inhibiting herbicides may not only affect algal community structure, but also provide cyanobacteria founder populations a window for dominance and thus play an important role in promoting cyanobacteria blooms.     

The use of glyphosate as the sole source of phosphorus or carbon for the selection of soil-borne fungal strains capable to degrade this herbicide

The herbicide glyphosate was used as a selection agent for isolation of fungal strains capable to degrade phosphorus-to-carbon bond from standard sandy-clay soil. The studies have shown that the herbicide used in Martin medium as a sole source of phosphorus br carbon caused the decrease of the fungal population and substantially changed strain composition, thus selecting those which are able to degrade glyphosate.     

Stress reactions in Vitis vinifera L. following soil application of the herbicide flumioxazin

In order to evaluate the stress effects of flumioxazin (fmx) on grapevine, a non-target plant (Vitis vinifera L.), physiological parameters such as carbohydrate content, water status or nitrogenous metabolites were investigated on fruiting cuttings and plants grown in vineyard. In the leaves of cuttings, the soil-applied herbicide induced stress manifestations including a decrease of the dry weight percentage and the soluble carbohydrate content during the first week after treatment. Thereafter, a decrease of the osmotic potential was observed, as well as a decrease of total protein content and a parallel accumulation of free amino acids, including proline. Altogether, these results suggest that soil-applied fmx induced a stress in grapevines, leading to leaf proteolysis. However, this stress was partially recovered 3 weeks after herbicide application, suggesting that the cuttings were capable to adapt to the fmx exposure. In the vineyard, the flumioxazin effects were still significant 5 months after the treatment, particularly in the CH cv. They included a decrease of the leaf dry weight percentage and soluble carbohydrate content, as well as an increase of the osmotic potential. The decrease of leaf soluble carbohydrates may have dramatic consequences for the berry growth and the reserve constitution. Moreover, treated plants were characterized by a decrease of the free amino acid content and an accumulation of ammonium, while the protein level did not significantly increase, suggesting a degradation of amino acids. The alteration of carbon and nitrogen status after herbicide treatment may affect the grapevine vigour in a long term.     

Population dynamics of Digitaria spp submitted to selection pressure by herbicides in sugarcane crop

The objective of this research was to study population dynamics of the weed crabgrass, genus Digitaria, submitted to selection pressure by herbicides currently applied in sugarcane crops in Brazil. In the first experiment two crabgrass species (Digitaria nuda and Digitaria ciliaris) and eight herbicide treatments applied in preemergence were used, and control percentage was evaluated at 7, 14, and 21 days after herbicide application (DAA). In the second experiment the level of tolerance through dose-response curve was determined for the species D. nuda and D. ciliaris, to the herbicides imazapyr, tebuthiuron, ametryne, and metribuzin. All the herbicides studied were efficient in controlling D. ciliaris, however, for D. nuda the best results were obtained only with ametryne, metribuzin, and isoxaflutole. The relation (T/S) between the rate required to reduce plant dry biomass (GR50) at 21 DAA of D. nuda and D. ciliaris was 16 for imazapyr and 6.3 for tebuthiuron, showing differential susceptibility of species; however for ametryne the rate T/S of 1.1 showed that D. nuda was not tolerant to this herbicide. For metribuzin, at 1.92 kg a.i. ha(-1), reduction of dry biomass was 80 and 90% to D. nuda and D. ciliaris, respectively. Even being controlled by metribuzin, D. nuda presented a higher level of tolerance to this herbicide, what was confirmed by the relationship T/S 14.4. As general conclusion of the research, it can be stated that the species D. nuda is more tolerant to ALS inhibiting herbicides and substituted urea, when compared with D. ciliaris; probably, D. nuda was selected by repetitive use of these herbicides.