Roundup Research Articles

Abstract P168: The Unholy Link: The Herbicide Glyphosate Contributes to Hypertension, Gut Microbiota Dysbiosis, and Sympathetic Overdrive

Introduction: Glyphosate, marketed as Roundup®, is a widely used potent herbicide in the US. Glyphosate inhibits 5-enolpyruvylshikimate-3-phosphate synthase of the shikimate pathway, a non-mammalian signaling pathway present in plants and microbes. Therefore, glyphosate has been deemed safe for humans. However, humans are holobionts with symbiotic gut microbiota susceptible to gut dysbiosis when exposed to glyphosate. Because gut microbiota is an important contributor to hypertension (HTN), we hypothesized that exposure to glyphosate leads to gut dysbiosis and HTN. Methods: Dahl salt-sensitive (S) rats were administered either vehicle or glyphosate (175 mg/kg/day) in drinking water for three weeks ad libitum. In one group of rats (N=4/group), sympathetic nerve activity was assessed using in situ whole nerve recordings and by measuring circulating norepinephrine (NE) levels by ELISA. In separate rats (N=6–8/group), blood pressure (BP) was measured once a week for three weeks by radiotelemetry, and circulating NE was determined by ELISA. At the endpoint, fecal microbiome analyses were performed using the Illumina and Oxford Nanopore platforms. Results: Exposure to glyphosate for just one week was sufficient to significantly increase systolic (173 mmHg vs 160 mmHg, p<0.0001) and mean arterial pressure (MAP) (148 mmHg vs 141 mmHg, p<0.0002), but not diastolic BP of S rats. Through week three, BP continued to rise in glyphosate-treated rats with greater differences in the systolic (195 mmHg vs 176 mmHg, p<0.0001), diastolic (142 mmHg vs 135 mmHg, p<0.0001), and MAP (168 mmHg vs 155 mmHg, p<0.0001). This was accompanied by increased sympathetic nerve activity and serum NE as early as one week of exposure to glyphosate (5.85 ng/mL vs 4.77 ng/mL, p<0.001), which continued at week three of glyphosate exposure (13.31 ng/mL vs 6.16 ng/mL, p<0.001), indicative of adrenergic overdrive. 16S data showed that glyphosate altered the composition of commensal gut microbiota, reducing bacterial α-diversity (Shannon entropy; p<0.05) and β-diversity (Bray-Curtis; p<0.02). Whole metagenomic analysis revealed a higher abundance of several bacterial species, including Candidatus spp., Erysipelaclostridium ramosum , and Clostridium sporogenes in the glyphosate-treated rats. Conclusions: Glyphosate exposure elevated BP, increased sympathetic nerve activity, and altered gut microbiota composition. This is the first proof-of-concept study to demonstrate that glyphosate contributes to HTN.

Climatic fluctuations alter the preference of stingless bees (Apidae, Meliponini) towards food contaminated with acephate and glyphosate

The global decline of pollinators has become a major concern for the scientific community, policymakers, and the general public. Among the main drivers of diminishing bee populations is the widespread use of agrochemicals. To gain a comprehensive understanding of the foraging dynamics of bees at agrochemical-contaminated areas, it is essential to consider both environmental conditions and the specific foraging ecology of bee species. For the first time, we conducted a semi-field study to investigate whether stingless bees exhibit a preference for food contaminated with agrochemicals compared to non- contaminated food, under natural weather conditions. Colonies of Plebeia lucii Moure, 2004 were placed in a greenhouse and subjected to a preference test, where bees were given the freedom to choose between contaminated or non-contaminated food sources following a preliminary training period. Within the greenhouse, we placed feeders containing realistic concentrations of an insecticide (acephate: 2 mg a.i./L), a herbicide (glyphosate: 31.3 mg a.i./L), or a mixture of both, alongside non-contaminated food. Environmental variables (temperature, humidity, and light intensity) were monitored throughout the experiment. At higher temperatures, the foragers preferred food containing the mixture of both agrochemicals or uncontaminated food over the other treatments. At lower temperatures, by contrast, the bees preferred food laced with a single agrochemical (acephate or glyphosate) over uncontaminated food or the agrochemical mixture. Our findings indicate that agrochemical residues in nectar pose a significant threat to P. lucii colonies, as foragers do not actively avoid contaminated food, despite the detrimental effects of acephate and glyphosate on bees. Furthermore, we demonstrate that even minor, natural fluctuations in environmental conditions can alter the colony exposure risk. Despite the interplay between temperature and bees' preference for contaminated food, foragers consistently collected contaminated food containing both agrochemicals, whether isolated or in combination, throughout the whole experiment.

Physiological responses of the stingless bee Partamona helleri to oral exposure to three agrochemicals: impact on antioxidant enzymes and hemocyte count.

Agrochemicals pose significant threats to the survival of bees, yet the physiological impacts of sublethal doses on stingless bees remain poorly understood. This study investigated the effects of acute oral exposure to three commercial formulations of agrochemicals [CuSO4 (leaf fertilizer), glyphosate (herbicide), and spinosad (bioinsecticide)] on antioxidant enzymes, malondialdehyde content (MDA), nitric oxide (NO) levels, and total hemocyte count (THC) in the stingless bee Partamona helleri. Foragers were exposed to lethal concentrations aimed to kill 5% (LC5) of CuSO4 (120μgmL-1) or spinosad (0.85μgmL-1) over a 24-h period. Glyphosate-exposed bees received the recommended label concentration (7400μgmL-1), as they exhibited 100% survival after exposure. Ingestion of CuSO4 or glyphosate-treated diets by bees was reduced. Levels of NO and catalase (CAT) remained unaffected at 0h or 24h post-exposure. Superoxide dismutase (SOD) activity was higher at 0h compared to 24h, although insignificantly so when compared to the control. Exposure to CuSO4 reduced glutathione S-transferase (GST) activity at 0h but increased it after 24h, for both CuSO4 and glyphosate. MDA levels decreased after 0h exposure to CuSO4 or spinosad but increased after 24h exposure to all tested agrochemicals. THC showed no difference among glyphosate or spinosad compared to the control or across time. However, CuSO4 exposure significantly increased THC. These findings shed light on the physiological responses of stingless bees to agrochemicals, crucial for understanding their overall health.

Association of urinary glyphosate levels with iron homeostasis among a representative sample of US adults: NHANES 2013–2018

PurposeGlyphosate and glyphosate-based herbicides (GBH), widely used globally, were initially considered harmless to humans. Experimental studies have suggested that these substances can disrupt iron homeostasis by interfering with iron uptake or triggering inflammatory responses. However, their potential impact on human iron homeostasis remains underexplored. Approach and resultsWe analyzed data from 5812 participants aged three and older from the 2013 to 2018 NHANES. We investigated the relationships between urinary glyphosate levels, oral iron intake, and markers of iron homeostasis, including serum iron, unsaturated iron-binding capacity (UIBC), total iron-binding capacity (TIBC), transferrin saturation, ferritin, and transferrin receptor. Higher urinary glyphosate levels were positively associated with oral iron intake (β = 1.310, S.E. = 0.382, P = 0.001). A one-unit increase in the natural logarithm (ln)-glyphosate was associated with lower serum iron (β = − 4.236, 95 % CI = − 6.432 to − 2.039, P < 0.001) and ferritin (β = − 9.994, 95 % CI = − 17.342 to − 2.647, P = 0.009), and higher UIBC (β = 5.431, 95 % CI = 1.061–9.800, P = 0.018) and transferrin receptor levels (β = 0.139, 95 % CI = 0.015–0.263, P = 0.029). Increasing glyphosate exposure was associated with significant decreases in serum iron and ferritin across exposure quintiles (trend P-values = 0.003 and 0.018, respectively). ConclusionsHigher glyphosate exposure is associated with reduced iron availability, suggesting potential disruptions in iron absorption. These findings underscore the need for further research into the health implications of glyphosate exposure on iron homeostasis.

Serum N-Glycan Changes in Rats Chronically Exposed to Glyphosate-Based Herbicides.

Glyphosate, the active ingredient in many herbicides, has been widely used in agriculture since the 1970s. Despite initial beliefs in its safety for humans and animals due to the absence of the shikimate pathway, recent studies have raised concerns about its potential health effects. This study aimed to identify glycomic changes indicative of glyphosate-induced toxicity. Specifically, the study focused on profiling N-glycosylation, a protein post-translational modification increasingly recognized for its involvement in various disorders, including neurological conditions. A comprehensive analysis of rat serum N-glycomics following chronic exposure to glyphosate-based herbicides (GBH) was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results revealed significant changes in the N-glycan profile, particularly in sialylated and sialofucosylated N-glycans. The analysis of N-glycans across gender subgroups provided insights into gender-specific responses to GBH exposure, with the male rats exhibiting a higher susceptibility to these N-glycan changes compared to females. The validation of significantly altered N-glycans using parallel reaction monitoring (PRM) confirmed their expression patterns. This study provides novel insights into the impact of chronic GBH exposure on serum N-glycan composition, with implications for assessing glyphosate toxicity and its potential neurological implications.

Changes in bumblebee queen gut microbiotas during and after overwintering diapause.

Bumblebees are key pollinators with gut microbiotas that support host health. After bumblebee queens undergo winter diapause, which occurs before spring colony establishment, their gut microbiotas are disturbed, but little is known about community dynamics during diapause itself. Queen gut microbiotas also help seed worker microbiotas, so it is important that they recover post-diapause to a typical community structure, a process that may be impeded by pesticide exposure. We examined how bumblebee queen gut microbiota community structure and metabolic potential shift during and after winter diapause, and whether post-diapause recovery is affected by pesticide exposure. To do so, we placed commercial Bombus impatiens queens into diapause, euthanizing them at 0, 2 and 4 months of diapause. Additionally, we allowed some queens to recover from diapause for 1 week before euthanasia, exposing half to the common herbicide glyphosate. Using whole-community, shotgun metagenomic sequencing, we found that core bee gut phylotypes dominated queen gut microbiotas before, during and after diapause, but that two phylotypes, Schmidhempelia and Snodgrassella, ceased to be detected during late diapause and recovery. Despite fluctuations in taxonomic community structure, metabolic potential remained constant through diapause and recovery. Also, glyphosate exposure did not affect post-diapause microbiota recovery. However, metagenomic assembly quality and our ability to detect microbial taxa and metabolic pathways declined alongside microbial abundance, which was substantially reduced during diapause. Our study offers new insights into how bumblebee queen gut microbiotas change taxonomically and functionally during a key life stage and provides guidance for future microbiota studies in diapausing bumblebees.

Perseverance of management is needed – Efficient long-term strategy of Reynoutria management

One of the most problematic invasive species in Europe are knotweeds from genus Reynoutria (Fallopia) which have significant negative impact on the native communities as well on human activities. Therefore, they are a target of many control programmes. Due to their high regeneration potential, their management is problematic, and only chemical treatment is reported to be sufficiently effective. The aim of this paper was to describe and analyse the patterns of Reynoutria invasion under long-term chemical treatment with glyphosate-based herbicide in The Morávka river floodplain, Czech Republic. The data covers 17 years of management which started with the European project “Preservation of alluvial forest habitats in the Morávka river basin”. We focus on (i) assessment of Reynoutria distribution during long-term management, (ii) analysis of the change of distribution according to the habitat, and (iii) discussion of the optimal management strategy based on the long-term data. Distribution data was obtained using GNSS field mapping. Before the start of the study in 2007, Reynoutria stands covered 29% of the study area (96.9 ha). As a result of systematic whole area chemical management, the extent decreased to 19.6% (65.3 ha) in 2009, and even reached 14.5% (48.2 ha) in 2013, three years after its end. Due to implementation of local chemical management in the following years, the area of Reynoutria was maintained at similar level, with minimum value 41.8 ha in 2018 and a slight increase in recent mapping in 2023. Beside the extent, the structure and coverage of invaded sites was analysed. There was a clear trend of fragmentation of larger polycormons with high coverage into many smaller and less dense ones as a result of chemical spraying. The average size of Reynoutria stand decreased from 0.61 ha in 2007 to half in 2013 (0.32 ha) to 0.15 ha in 2023. Testing of the effects of time, habitat, and biotope did not reveal significant differences of changes of extent and abundance over different environments (forest, open, bare ground), which indicates that there are no differences in reaction to management in the studied habitat and vegetation types. Our study provides a robust and unique overview of the invasion, reinvasion, and suppression dynamics for an important invasive species. If herbicide management is used, chemical treatment must be quite long-term as even three years of intensive glyphosate foliar spray application was not sufficient for the complete eradication of Reynoutria. Therefore, we propose the following procedure for effective chemical management of Reynoutria: 1) In largely infested sites, the first step is to reduce the distribution of Reynoutria stands to isolated polycormons. This phase can last 3–5 years. 2) After reaching the state of sparse distribution of Reynoutria, we recommend herbicide application only in periods of every 3–5 years depending on the local context and rate of regrowth. 3) At sites exposed to soil disturbances, where the soil is contaminated by fragments of Reynoutria rhizomes, there is a need to apply herbicide immediately to target newly resprouting individuals.

Evolving dual-trait EPSP synthase variants using a synthetic yeast selection system

The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) functions in the shikimate pathway which is responsible for the production of aromatic amino acids and precursors of other essential secondary metabolites in all plant species. EPSPS is also the molecular target of the herbicide glyphosate. While some plant EPSPS variants have been characterized with reduced glyphosate sensitivity and have been used in biotechnology, the glyphosate insensitivity typically comes with a cost to catalytic efficiency. Thus, there exists a need to generate additional EPSPS variants that maintain both high catalytic efficiency and high glyphosate tolerance. Here, we create a synthetic yeast system to rapidly study and evolve heterologous EPSP synthases for these dual traits. Using known EPSPS variants, we first validate that our synthetic yeast system is capable of recapitulating growth characteristics observed in plants grown in varying levels of glyphosate. Next, we demonstrate that variants from mutagenesis libraries with distinct phenotypic traits can be isolated depending on the selection criteria applied. By applying strong dual-trait selection pressure, we identify a notable EPSPS mutant after just a single round of evolution that displays robust glyphosate tolerance (Ki of nearly 1 mM) and improved enzymatic efficiency over the starting point (~2.5 fold). Finally, we show the crystal structure of corn EPSPS and the top resulting mutants and demonstrate that certain mutants have the potential to outperform previously reported glyphosate-resistant EPSPS mutants, such as T102I and P106S (denoted as TIPS), in whole-plant testing. Altogether, this platform helps explore the trade-off between glyphosate resistance and enzymatic efficiency.

Greenhouse study and interviews indicate glyphosate residue via feed-feces-fertilizer route is a risk for horticultural producers using manure-based fertilizer

BackgroundThe herbicide glyphosate is the most widely used active ingredient in pesticides globally. Residues have been found in people, livestock, food and animal feed, and in the environment, but little is known about glyphosate residue in manure-based fertilizer. We describe a feed-feces-fertilizer route of glyphosate contamination with negative impacts for horticultural production. This exposure can harm sensitive plants, such as tomato, and pose a risk to effective waste disposal and nutrient cycling along principles of the circular economy.We review the use and history of glyphosate and present a mixed methods research based on a real-world case from Finland where glyphosate residue in poultry manure fertilizer was suspected of inhibiting commercial organic tomato production. To test the fertilizer, we grew 72 ‘Encore’ variety tomato plants for 14 weeks in a climate-controlled greenhouse according to the practices of the commercial grower. To ascertain awareness and potential contamination mitigation measures, we contacted five fertilizer companies with sales of biogenic fertilizer in Finland, two farming organizations, a feed company, and two government organizations working on nutrient cycling and agricultural circular economy.ResultsThe total harvest of tomatoes grown with fertilizer with the higher content of glyphosate residue was 35% smaller and the yield of first-class tomatoes 37% lower than that of the control, with lower glyphosate concentration. Two of the five fertilizer companies identified poultry manure as a source of glyphosate contamination. Companies with awareness of pesticide residues reported interest in establishing parameters for pesticide residues.ConclusionsThe extent of glyphosate contamination of recycled fertilizers is unknown, but this study shows that such contamination occurs with negative impacts on crop production. Lack of testing and regulation to ensure that recycled fertilizers are free from harmful levels of glyphosate or other pesticides creates risks for agricultural producers. The issue is particularly acute for certified organic producers dependent on these products, but also for sustainable transitions away from mineral fertilizers in conventional farming. The example from Finland shows that a model of co-production between fertilizer producers and state regulatory agencies to establish safe limits can benefit both fertilizer producers and their customers.

Potential Role of Glyphosate, Glyphosate-Based Herbicides, and AMPA in Breast Cancer Development: A Review of Human and Human Cell-Based Studies.

The potential connection between exposure to glyphosate and glyphosate-based herbicides (GBHs) and breast cancer risk is a topic of research that is rapidly gaining the public's attention due to the conflicting reports surrounding glyphosate's potential carcinogenicity. In this review, we synthesize the current published biomedical literature works that have explored associations of glyphosate, its metabolite, aminomethylphosphonic acid (AMPA), and GBHs with breast cancer risk in humans and human cell-based models. Using PubMed as our search engine, we identified a total of 14 articles that were included in this review. In the four human studies, urinary glyphosate and/or AMPA were associated with breast cancer risk, endocrine disruption, oxidative stress biomarkers, and changes in DNA methylation patterns. Among most of the 10 human cell-based studies, glyphosate exhibited endocrine disruption, induced altered gene expression, increased DNA damage, and altered cell viability, while GBHs were more cytotoxic than glyphosate alone. In summary, numerous studies have shown glyphosate, AMPA, and GBHs to have potential carcinogenic, cytotoxic, or endocrine-disruptive properties. However, more human studies need to be conducted in order for more definitive and supported conclusions to be made on their potential effects on breast cancer risk.

Allosteric Regulation of Pyruvate Kinase Enables Efficient and Robust Gluconeogenesis by Preventing Metabolic Conflicts and Carbon Overflow.

Glycolysis and gluconeogenesis are reciprocal metabolic pathways that utilize different carbon sources. Pyruvate kinase catalyzes the irreversible final step of glycolysis, yet the physiological function of its regulation is poorly understood. Through metabolomics and enzyme kinetics studies, we discovered that pyruvate kinase activity is inhibited during gluconeogenesis in the soil bacterium Bacillus subtilis . This regulation involves an extra C-terminal domain (ECTD) of pyruvate kinase, which is essential for autoinhibition and regulation by metabolic effectors. Introducing a pyruvate kinase mutant lacking the ECTD into B. subtilis resulted in defects specifically under gluconeogenic conditions, including inefficient carbon utilization, slower growth, and decreased resistance to the herbicide glyphosate. These defects are not caused by the phosphoenolpyruvate-pyruvate-oxaloacetate futile cycle. Instead, we identified two significant metabolic consequences of pyruvate kinase dysregulation during gluconeogenesis: increased carbon overflow into the medium and failure to expand glycolytic intermediates such as phosphoenolpyruvate (PEP). In silico analysis revealed that in wild-type cells, an expanded PEP pool enabled by pyruvate kinase regulation is critical for the thermodynamic feasibility of gluconeogenesis. Our findings underscore the importance of allosteric regulation during gluconeogenesis in coordinating metabolic flux, efficient energy utilization, and antimicrobial resistance.

A Pilot Study to Evaluate Exposure to Polyoxyethylene Tallow Amine (POEA), a Glyphosate Formulation Adjuvant, in a US pregnant population

A Pilot Study to Evaluate Exposure to Polyoxyethylene Tallow Amine (POEA), a Glyphosate Formulation Adjuvant, in a US pregnant population

Vitamin E alleviates glyphosate-based herbicide-induced progesterone secretion inhibition and oxidative stress increase in chicken primary granulosa cells

Glyphosate-based herbicides (GBH) are the most extensively used herbicides worldwide. Despite a presumed nondangerousness for animals, several studies reported negative effects after a GBH exposure in several animal models including birds, notably on reproductive functions. Several studies concerning the advantages of Vitamin E (VE) for antioxidant activity but also growth and reproduction have been reported in birds. However, it remains unclear whether VE could alleviate the negative effect of GBHs on chicken ovarian cells. Here we exposed chicken primary granulosa cells (GCs) from F1 and F3/4 follicles to growing doses of GBH (0.036, 0.36, 3.6, and 36 gly eq/L), with or without VE supplementation (1 mg/L) and investigated cell viability, proliferation, oxidative stress and steroidogenesis. GBH exposure did not affect F1 and F3 GCs viability but it increased cell proliferation only in F1 GCs and this effect was not altered by VE. In both F1 and F3/4 GCs, GBH exposure increased total oxidant status (TOS), reduced total antioxidant status (TAS) and consequently increased index of oxidative stress (OSI) in dose dependent manner. This latter effect for GBH 36 mg eq gly/L was totally abolished in response to VE. In both F1 and F3/4 GCs, GBH exposure reduced progesterone secretion in a dose dependent manner and this effect with GBH 0.36 and 1.8 mg eq glyphosate/L was alleviated by VE. However, we did not observe any effect of GBH and VE on the gene expression of several components of the steroidogenesis process. Taken together, these results show that GBH may have endocrine disruptor effects, and that these effects might be alleviated by antioxidant VE supplementation.

Perinatal Exposure to Glyphosate or a Commercial Formulation Alters Uterine Mechanistic Pathways Associated with Implantation Failure in Rats.

Perinatal exposure to a glyphosate-based herbicide (GBH) or its active ingredient, glyphosate (Gly), has been demonstrated to increase implantation failure in rats. This study investigates potential mechanisms of action, analyzing uterine preparation towards the receptive state. Pregnant Wistar rats (F0) were treated orally with GBH or Gly (3.8 and 3.9 mg Gly/kg/day, respectively) from gestational day (GD) 9 until weaning. Adult F1 females became pregnant and uterine samples were collected on GD5 (preimplantation period). Histomorphological uterine parameters were assessed. Immunohistochemistry was applied to evaluate cell proliferation and protein expression of estrogen receptors (ERα and ERβ), cell cycle regulators (PTEN, cyclin G1, p27, and IGF1R-α), and the Wnt5a/β-catenin/FOXA2/Lif pathway. Both GBH and Gly females showed increased stromal proliferation, associated with a high expression of ERs. Dysregulation of PTEN and cyclin G1 was also observed in the Gly group. Reduced gland number was observed in both groups, along with decreased expression of Wnt5a/β-catenin/FOXA2/Lif pathway in the glandular epithelium. Overall, GBH and Gly perinatal exposure disrupted intrinsic uterine pathways involved in endometrial proliferation and glandular function, providing a plausible mechanism for glyphosate-induced implantation failure by compromising uterine receptivity. Similar effects between GBH and Gly suggest the active principle mainly drives the adverse outcomes.

Decreased activity in zebrafish larvae exposed to glyphosate-based herbicides during development-potential mediation by glucocorticoid receptor.

Glyphosate-based herbicides (GBH) are a widely used group of pesticides that have glyphosate (GLY) as main active compound and are used to control a wide range of weeds. Experimental and epidemiological studies point to neurotoxicity and endocrine disruption as main toxic effects. The aim of this study was to investigate the effects of developmental exposure to GLY and GBH on locomotor behavior, and the possible contribution of GR-mediated signaling. We used zebrafish (Danio rerio) larvae in a continuous exposure regimen to GLY or GBH in the rearing medium. Alongside TL wildtype, we used a mutant line carrying a mutation in the GR which prevents the GR from binding to DNA (grs357), as well as a transgenic strain expressing a variant of enhanced green fluorescent protein (d4eGFP) controlled by a promoter carrying multiple GR response elements (SR4G). We found that acute exposure to GBH, but not GLY, activates GR-mediated signaling. Using a continuous developmental exposure regime, we show that wildtype larvae exposed to GBH display decreased spontaneous activity and attenuated response to environmental stimuli, a pattern of alteration similar to the one observed in grs357 mutant larvae. In addition, developmental exposure to GBH has virtually no effects on the behavior of grs357 mutant larvae. Taken together, our data indicate that developmental exposure to GBH has more pronounced effects than GLY on behavior at 5 dpf, and that interference with GR-mediated signaling may have a relevant contribution.

Soil degradation and herbicide pollution by repeated cassava monoculture within Thailand's conservation region.

In a national park in Northeast Thailand, agricultural land has been converted from natural forest by small-scale farmers for cassava agriculture. We hypothesise that long-termed cassava monoculture leads to the degradation of soil properties. To test the hypothesis, we conducted a five-year (2016-2020) study on the physical and chemical properties of soil in cassava farmland, and also examined the soil properties of its adjacent natural forests, as a control. The examined cassava farmland was converted from the natural forest during the five years from 2011 to 2015. The significant decrease in organic carbon and the increases in exchangeable potassium and bulk density were found in 2016, indicating that these soil properties varied quickly following the farmland conversion. On the other hand, the significant increase in soil nitrogen and the decrease in pH were found later in 2020, indicating that these soil properties were gradually altered by repeated agricultural activities, such as fertilizer application and trampling. In contrast, there were no significant differences in available phosphate, electrical conductivity, cation exchange capacity, and the soil texture (the fractions of sand, silt, and clay) among the forest and farmland soils. The cation exchange capacity was positively correlated to the fraction of clay, the organic carbon, and pH. The use of glyphosate and paraquat herbicides is prohibited within national parks in Thailand. However, in 2020, glyphosate was detected in farmland soil (up to 5.0 mg kg-1) during both the rainy and dry seasons, and glyphosate (up to 2.5 mg l-1) was detected in stream water from the farmland during the dry season at least in 2020. Soil degradation and herbicide pollution may carry a high risk of causing irreversible changes in terrestrial ecosystems. We discuss the root causes of this issue from perspectives of agricultural production, economy, and the environmental impact, and propose effective policy measures.

Early exposure to glyphosate during larval development induces late behavioural effects on adult honey bees

As the most abundant pollinator insect in crops, Apis mellifera is a sentinel species of the pollinator communities. In these ecosystems, honey bees of different ages and developmental stages are exposed to diverse agrochemicals. However, most toxicological studies analyse the immediate effects during exposure. Late effects during adulthood after early exposure to pollutants during larval development are poorly studied in bees. The herbicide glyphosate (GLY) is the most applied pesticide worldwide. GLY has been detected in honey and beebread from hives near treated crops. Alterations in growth, morphogenesis or organogenesis during pre-imaginal development could induce late adverse effects after the emergence. Previous studies have demonstrated that GLY alters honey bee development, immediately affecting survival, growth and metabolism, followed by late teratogenic effects. The present study aims to determine the late impact on the behaviour and physiology of adult bees after pre-imaginal exposure to GLY. For that, we reared brood in vitro or in the hive with sub-chronic exposure to the herbicide with the average detected concentration in hives. Then, all newly emerged bees were reared in an incubator until maturity and tested when they became nurse-aged bees. Three behavioural responses were assessed as markers of cognitive and physiological impairment. Our results show i) decreased sensitivity to sucrose regardless of the rearing procedure, ii) increased choice latency and locomotor alterations during chemotaxis and iii) impaired associative learning. These late toxicity signs could indicate adverse effects on task performance and colony efficiency.

Establishing native wildflower habitats in urban settings on a low budget

ABSTRACT This study aimed to compare the costs of two methods in establishing a native wildflower habitat using transplants while controlling weeds. The first method involved no-tillage and glyphosate herbicide and the second method used dazomet herbicide with tillage. Direct expenses included the initial establishment of the transplants under two differing soil treatments and were calculated separately. Expense differences between the two methods result from the prices of the two herbicides, type of equipment used in soil preparation and the type of tasks requiring labour. Results show a high rate (85%–90%) of the transplants becoming established regardless of the method used, suggesting that both may be considered suitable. With conservation and sustainability in mind, the cost estimates provide benchmarks for different methods of planting wildflower habitats in urban settings.

Connectivity mediates the spatial ecological impacts of a glyphosate-based herbicide in experimental metaecosystems

Metacommunity ecology has shown that connectivity is important for the persistence of a species locally and across connected ecosystems, however we do not know if ecological effects in freshwater ecosystems exposed to biocides leaking from agriculture depend on metaecosystem connectivity. We experimentally replicated metaecosystems in the laboratory using gradostats as a model system. We tested the effects of connectivity, in terms of node distance from the pollutant-source, flow rate, and a glyphosate-based herbicide, on phytoplankton productivity, diversity and stability. Gradostats were composed of interconnected equally spaced nodes where resources and phytoplankton move directionally along a gradient of increasing distance from the source of the polluting herbicide. We hypothesised that ecological effects would be stronger in the node situated closer to the point of herbicide input, but that flow would suppress phytoplankton populations in distant nodes. Overall, RoundUp impacted phytoplankton productivity and stability by reducing algal biomass and abundances. This occurred especially in the node closest to the diluted herbicide point-source and under high flow, where species abundances were heavily suppressed by the effects of the rapidly flowing herbicide. At low flow on the other hand, distant nodes where buffered from the effects of the slow-moving herbicide. No differences in beta and gamma diversity among replicate metaecosystems was found; however, a significant loss of alpha diversity in all metaecosystems occurred through time until the end of the experiment. Together, these results point to the importance of considering aquatic connectivity in management plans for monitoring and mitigating unintended ecological consequences of agrochemical runoff.

Assessment of cytotoxic and genotoxic effects of glyphosate-based herbicide on glioblastoma cell lines: Role of p53 in cellular response and network analysis

Glyphosate, the world's most widely used herbicide, has a low toxicity rating despite substantial evidence of adverse health effects. Furthermore, glyphosate-based formulations (GBFs) contain several other chemicals, some of which are known to be harmful. Additionally, chronic, and acute exposure to GBFs among rural workers may lead to health impairments, such as neurodegenerative diseases and cancer. P53 is known as a tumor suppressor protein, acting as a key regulator of the cellular response to stress and DNA damage. Therefore, mutations in the TP53 gene, which encodes p53, are common genetic alterations found in various types of cancer. Therefore, this study aimed to evaluate the cytotoxicity and genotoxicity of GBF in two glioblastoma cell lines: U87MG (TP53-proficient) and U251MG (TP53-mutant). Additionally, the study aimed to identify the main proteins involved in the response to GBF exposure using Systems Biology in a network containing p53 and another network without p53. The MTT assay was used to study the toxicity of GBF in the cell lines, the clonogenic assay was used to investigate cell survival, and the Comet Assay was used for genotoxicity evaluation. For data analysis, bioinformatics tools such as String 12.0 and Stitch 5.0 were applied, serving as a basis for designing binary networks in the Cytoscape 3.10.1 program. From the in vitro test analyses, it was observed a decrease in cell viability at doses starting from 10 ppm. Comet Assay at concentrations of 10 ppm and 30 ppm for the U251MG and U87MG cell lines, respectively observed DNA damage. The network generated with systems biology showed that the presence of p53 is important for the regulation of biological processes involved in genetic stability and neurotoxicity, processes that did not appear in the TP53-mutant network.