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Atrazine and the hypothalamo-pituitary-gonadal axis in sexually maturing precocial birds: studies in male Japanese quail.

The herbicide atrazine is a putative endocrine disruptor. The present studies investigated the effects of atrazine in male Japanese quail during sexual maturation. Atrazine was administered for two weeks in the diet or systemically to birds under long photoperiods. Atrazine had no effect on mortality but depressed both feed intake and growth (average daily gain [ADG] in g/day) at dietary concentrations of 1000 ppm. Atrazine in the diet at 10 ppm, but at no other concentrations, increased testes weight and gonadal-somatic-index and decreased the seminiferous tubule diameter-to-testis weight ratio. However, there were no effects on absolute tubule diameter, relative stage of testicular development, or the presence of a lumen. Atrazine in the diet at 1000 ppm increased circulating concentrations of testosterone but this effect was not observed consistently in all studies. Dietary atrazine at 10 ppm increased circulating concentrations of estradiol. Moreover, in one study, atrazine at 1000 ppm in the diet decreased circulating concentrations of luteinizing hormone. Atrazine administered systemically exerted no effect on indices of growth or reproduction. Atrazine did not mimic the effects of either estradiol or tamoxifen in male quail; thus, atrazine did not exhibit overt estrogenic or anti-estrogenic activity. Conversely, atrazine augmented the effects of testosterone and estradiol on testis regression, presumably by increasing the negative-feedback effects of these sex-steroids on follicle stimulating hormone secretion. It is concluded that atrazine up to 1000 ppm in the diet may exert some effects on reproductive development in sexually maturing male birds, but these are inconsistent and modest.

Animals↗

Biological granulated activated carbon fluidized bed reactor for atrazine remediation.

To show that an adsorbing biofilm carrier (GAC) can be advantageous for atrazine bioremediation over a non-adsorbing carrier, fluidized bed (FB) reactors were operated under atrazine limiting concentrations using Pseudomonas sp. strain ADP as the atrazine degrading bacteria. The following interrelated subjects were investigated: 1) atrazine adsorption to GAC under conditions of atrazine partial penetration in the biofilm, 2) differences in atrazine degradation rates and 3) stability of atrazine biodegradation under non-sterile anoxic conditions in the GAC reactor versus a reactor with a non-adsorbing biofilm carrier. Results from batch adsorption tests together with modeling best described the biofilm as patchy in nature with covered and non-biofilm covered areas. Under conditions of atrazine partial penetration in the biofilm, atrazine adsorption occurs in the non-covered areas and is consequently desorbed at the base of the biofilm substantially increasing the active biofilm surface area. The double flux of atrazine to the biofilm in the GAC reactor results in lower effluent atrazine concentrations as compared to a FB reactor with a non-adsorbing carrier. Moreover, under non-sterile denitrification conditions, atrazine degradation stability was found to be much higher (several months) using GAC as a biofilm carrier while non-adsorbing carrier reactors showed sharp deterioration within 30 days due to contamination of non-atrazine degrading bacteria.

Adsorption↗

Atrazine mineralization potential in two wetlands.

The fate of atrazine in agricultural soils has been studied extensively but attenuation in wetland systems has received relatively little attention. The purpose of this study was to evaluate the mineralization of atrazine in two wetlands in central Ohio. One was a constructed wetland, which is fed by Olentangy River water from an agricultural catchment area. The other was a natural fen (Cedar Bog) in proximity to atrazine-treated cornfields. Atrazine mineralization potential was measured by 14CO2 evolution from [U-ring-14C]-atrazine in biometers. The constructed wetland showed 70-80% mineralization of atrazine within 1 month. Samples of wetland water that were pre-concentrated 200-fold by centrifugation also mineralized 60-80% of the added atrazine. A high extent of atrazine mineralization (75-81% mineralized) was also associated with concentrated water samples from the Olentangy River that were collected upstream and downstream of the wetland. The highest levels of mineralization were localized to the top 5 cm zone of the wetland sediment, and the activity close to the outflow at the Olentangy wetland was approximately equal to that near the inflow. PCR amplification of DNA extracted from the wetland sediment samples showed no positive signals for the atzA gene (atrazine chlorohydrolase), while Southern blots of the amplified DNA showed positive bands in five of the six Olentangy wetland sediment samples. Amplification with the trzD (cyanuric acid amidohydrolase) primers showed a positive PCR signal for all Olentangy wetland sediment samples. There was little mineralization of atrazine in any of the Cedar Bog samples. DNA extracted from Cedar Bog samples did not yield PCR products, and the corresponding Southern hybridization signals were absent. The data show that sediment microbial communities in the Olentangy wetland mineralize atrazine. The level of activity may be related to the seasonality of atrazine runoff entering the wetland. Comparable activity was not observed in the Cedar Bog, perhaps because it does not directly receive agricultural runoff. Qualitatively, the detection of the genes was associated with measurable mineralization activity which was consistent with the differences between the two study sites.

Agriculture↗

Salicylate activity. 2. Potentiation of atrazine.

Atrazine [6-chloro-N-ethyl-N'-(1-methylethyl)-1,3,5-triazine-2,4-diamine] inhibits photosystem II (PSII) and is commonly used to control weeds in maize. It has been found that addition of sodium salicylate (sodium 2-hydroxybenzoate; NaSA) increased the postemergence herbicidal activity of atrazine against dicotyledonous weeds. NaSA also potentiated the activity of bentazon, another PSII-inhibiting herbicide. NaSA increased atrazine activity when applied either as a tank mix or up to 96 h prior to atrazine application. Other salicylates and the plant disease resistance inducers acibenzolar-S-methyl [benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester] and 2,6-dichloroisonicotinic acid also increased atrazine activity. Among the compounds tested, 3-chloro-5-fluorosalicylate, 4-chlorosalicylate, or 2,6-dichloroisonicotinic acid combined with atrazine yielded the greatest increase in herbicidal activity. Potentiation of atrazine by NaSA was greater at higher temperatures (35 and 25 > 15 degrees C). Also, greater potentiation was observed as the light level decreased. In darkness, NaSA alone or in combination with atrazine caused plant death, whereas atrazine alone had little effect. NaSA increased atrazine activity on npr1-2, an Arabidopsis mutant compromised in SA-induced disease resistance. Atrazine activity was also potentiated by NaSA on the ethylene insensitive mutant ein2-1. This indicates that atrazine potentiation is independent of either salicylate-induced disease resistance or ethylene perception.

Arabidopsis↗

Correspondence of salivary and plasma concentrations of atrazine in rats under variable salivary flow rate and plasma concentration.

The stability of the saliva/plasma (S/P) concentration ratio of atrazine was determined under varying conditions of salivary flow rate and plasma concentration of atrazine in Sprague-Dawley rats. In the salivary flow study, whole saliva samples were collected at different salivary flow rates while the plasma concentration of atrazine was maintained at a steady-state level of approximately 150 micrograms/L. In the plasma level study, whole saliva samples were collected at two steady-state plasma concentrations of atrazine (50 and 250 micrograms/L), while salivary flow rate was maintained at a relatively constant level. In both studies, atrazine concentrations in whole saliva and arterial plasma demonstrated a consistent relationship, but salivary concentrations were always lower than those of arterial plasma. Linear regression analysis demonstrated that the S/P concentration ratio of atrazine was not significantly different for salivary flow rates ranging from 23 to 92 microL/min/kg body weight, and did not vary for atrazine plasma concentrations between 30 and 433 micrograms/L. The S/P concentration ratio of atrazine was relatively constant throughout each experimental period (0.68 +/- 0.1 and 0.70 +/- 0.11 for salivary flow and plasma level studies, respectively) and did not differ significantly between rats. When data from both studies were pooled, salivary concentrations were highly correlated with plasma concentrations (r2 = .94). It is concluded that under these experimental conditions, the stability of the S/P concentration ratio of atrazine is not affected by variations in salivary flow rate or atrazine plasma concentrations. The results from this study support the conclusion that atrazine salivary concentrations can be used to predict plasma levels of atrazine in rats.

Analysis of Variance↗

Enhanced degradation of deethylatrazine in an atrazine-history soil of Iowa.

The degradation of deethylatrazine (DEA), a major metabolite of atrazine, was studied by using radiotracers in soils with two different atrazine histories. DEA degradation was enhanced in soils which had received long-term exposure to atrazine (atrazine-history soil) compared with soils that had not received long-term atrazine exposure (no-history soil). After 60 days of incubation, mineralization of DEA to 14CO2 in the atrazine-history surface soil was twice that in the no-history surface soils, with 34% and 17% of the applied 14C-DEA as CO2, respectively. In surface soils, 25% of the applied 14C remained as DEA in the atrazine-history soil, compared with 35% in the no-history soil. Microbial plate counts indicated an increase in numbers of bacteria and fungi in soils incubated with DEA compared to control soils. No significant difference in total microbial respiration was seen among atrazine-history and no-history soils incubated with DEA, but DEA-treated soils had greater microbial respiration than untreated control soils after 6 days. A 14C-most-probable-number procedure was used to enumerate specific DEA degraders. A greater number of DEA degraders were indicated in atrazine-history subsurface soil compared with all other soils in this study (p < 0.05). From this study, it appears that an increase in microbial activity contributes to decreased persistence and increased degradation of DEA in soils that have had long-term exposure to atrazine at field application rates, compared to soils with no long-term exposure. Decreased persistence of this major metabolite of atrazine in atrazine-history soils is important in that there will be less available for movement in surface runoffs.

Atrazine↗

Reproductive and developmental effects of atrazine on the estuarine meiobenthic copepod Amphiascus tenuiremis.

Atrazine is one of the most widely used herbicides in the United States. Atrazine concentrations in coastal environments chronically range from 90 ng/L to 46 microg/L, with rare but measured concentrations near 60 microg/L at edge-of-field conditions. Chronic atrazine effects on estuarine benthos exposed to environmentally relevant concentrations are unknown. The purpose of this research was to assess atrazine reproductive and developmental effects over multiple-generation exposures of the copepod Amphiascus tenuiremis. Copepods were chronically exposed to two environmentally relevant nominal atrazine concentrations (2.5 and 25 microg/L, and to an environmentally unrealistic concentration (250 microg/L). Chronic exposures were performed using a 96-well microplate life cycle bioassay. Individual stage I copepodites (C1, n = 60/treatment) were reared through two generations (F0 and F1) to sexual maturity and individually mated in microwells containing 200 microl of atrazine solution. Copepod survival across all treatments and generations was >95%. Atrazine did not affect development to reproductive maturity, time to egg extrusion, or time to egg hatch (p > 0.05). However, reproductive failures increased across generations with increasing atrazine concentrations. Reproductive failures in the 0-, 2.5-, 25-, and 250-microg/L atrazine treatments were 11, 11, 20, and 24% for the F0 and 4, 9, 26, and 38% for the F1, respectively. Compared to controls, total nauplii production per female was reduced by approximately 22% in F0 females exposed to 250 microg/L atrazine (p < 0.05), and by approximately 23%, approximately 27%, and approximately 32% in F1 females exposed to 2.5-, 25-, and 250-microg/L atrazine treatments, respectively (p < 0.05). The combined effect of reproductive failure and reduced offspring production significantly reduced total population growth in the F1 generation (p < 0.05) even at atrazine concentrations lower than that considered safe for seawater chronic exposure (26 microg/L).

Animals↗

Field history and dissipation of atrazine and metolachlor in Colorado.

Farmers in eastern Colorado have commented that atrazine does not provide the length of weed control that they expected in fields that have received multiple applications of the herbicide. Multiple laboratory studies suggest that atrazine dissipates more rapidly in soils with a history of atrazine use compared with soils that had not been treated with the herbicide and this could be related to the above observation. Field and laboratory studies were conducted to determine the rate of dissipation of atrazine and metolachlor in fields in Colorado. The published half-lives of atrazine and metolachlor are 60 and 56 d, respectively. In the field studies, the half-lives of atrazine and metolachlor in the top 15 cm of the soil ranged between 3.5 and 7.2 d and 17.9 and 18.8 d, respectively. In laboratory studies, the half-life of atrazine varied from 1.4 to 19.8 d with the shortest half-life occurring in soils which had been treated with atrazine for at least 5 yr. The longest half-life was in a soil that had never received atrazine. The half-life of metolachlor in these same soils varied from 10.6 to 28.2 d. There was no apparent relationship between the half-life of metolachlor and the half-life of atrazine in the laboratory studies. These results confirm farmers' observation of the shorter residual activity of atrazine in Colorado fields receiving atrazine over multiple years.

Acetamides↗

[Environmental characteristics of the atrazine in the waters in East Liaohe River Basin].

Based on the field investigation and the Laboratory experiment, and taking the Geographic Information System (GIS) techniques as the key data disposal platform, the spatial and temporal differences and the enrichment characteristics of atrazine in the surface water in East Liaohe River Basin were systematically analyzed, the effect factors on that also were interpreted. The results show that, the average atrazine content in waters in regions with glebe or without glebe were namely 9.71 microg x L(-1) and 8.854 microg x L(-1), and the lower reach of the trunk was formed as the focus filed of the atrazine in waters. July is the month that the water contained the largest atrazine with the maximum content of 18.93 microg x L(-1). The spatial pattern of the atrazine in the waters clearly shows that, the atrazine used in the glebe are endangering the surface waters quality in the whole basin and will worsen the waters environment of East Liaohe River basin. The normalized enrichment index of the atrazine in the surface water related to plant atrazine is 0.605-1.750 with the spatial difference, the right part of the trunk is the continuous region with high index, the left part of the trunk is on the contrary, and the lower reaches is the focus region with high index. The normalized enrichment index of the atrazine in the surface waters related to soil atrazine has the spatial difference same to the index related to soil, however, the region with large index or little index has the scattering features. With respect to the effect of soil on the atrazine in the water, the integrated token index of the low moor peat soil is the largest and the peat moor soil and the paddy soil are the least. With respect to effect of the landscape pattern on the atrazine in the surface waters, the integrated token index will increase with the interaction between patches. With a certain range, the integrated token index will increase with the precipitation and runoff; however, the integrated token index will decrease with precipitation if the precipitation excesses the range.

Atrazine↗

Isolation and characterisation of new Gram-negative and Gram-positive atrazine degrading bacteria from different French soils.

The capacity of 12 soils to degrade atrazine was studied in laboratory incubations using radiolabelled atrazine. Eight soils showed enhanced degradation of this compound. Twenty-five bacterial strains able to degrade atrazine were isolated by an enrichment method from 10 of these soils. These soils were chosen for their wide range of physico-chemical characteristics. Their history of treatment with atrazine was also variable. The genetic diversity of atrazine degraders was determined by amplified ribosomal restriction analysis (ARDRA) of the 16S rDNA gene with three restriction endonucleases. The 25 bacterial strains were grouped into five ARDRA types. By sequencing and aligning the 16S rDNA genes, the isolates were shown to belong to the Gram-negative species Chelatobacter heintzii, Aminobacter aminovorans, Stenotrophomonas maltophilia and to the Gram-positive genus Arthrobacter crystallopoietes. These species were not described previously as being capable of atrazine degradation. Most Gram-negative bacteria could mineralise (14)C ring labelled atrazine and carried the atzA, atzB, atzC and trzD genes. Gram-positive strains could convert atrazine to cyanuric acid and carried only the atzB and atzC genes. In this study, we describe the atrazine degradation capacities and corresponding genes in bacterial species that were not known as atrazine degraders. We report for the first time the occurrence of the trzD gene in these atrazine-mineralising bacteria and we demonstrate the potential use of colony hybridisation to isolate bacteria involved in atrazine degradation.

Journal Article↗

A risk assessment of atrazine use in California: human health and ecological aspects.

A risk assessment of the triazine herbicide atrazine has been conducted by first analyzing the toxicity database and subsequently estimating exposure. Margins of safety (MOS) were then calculated. Toxicity was assessed in animal studies and exposure was estimated from occupational and dietary sources. In acute toxicity studies, atrazine caused developmental toxicity in the rabbit [no observed effect level (NOEL) 5 mg kg(-1) day(-1)] and cardiotoxicity in a dog chronic study (NOEL 0.5 mg kg(-1) day(-1)); cancer (mammary glands) resulted from lifetime exposure. The mammary tumors, which occurred specifically in female Sprague-Dawley rats, were malignant, increased in a dose-dependent manner and were also observed with other, related triazines. Evidence for a genotoxic basis for these tumors was either equivocal or negative. Triazines have been shown to be clastogenic in Chinese hamster ovary cells, in vitro, but without showing a convincing dose/response relationship. Atrazine can be converted into genotoxic N-nitrosoatrazine in the environment or the digestive system, suggesting that N-nitrosamines derived from triazines could be oncogenic. However, it was concluded that N-nitrosotriazines are unlikely to play a significant role in triazine-induced rat mammary gland tumors. An endocrine basis for the mammary tumors, involving premature aging of the female SD rat reproductive system, has been proposed. A suppression of the luteinizing hormone surge during the estrus cycle by atrazine leads to the maintenance of elevated blood levels of 17beta-estradiol (E2) and prolactin. The mechanism for tumor development may include one or more of the following: the induction of aromatase (CYP19) and/or other P450 oxygenases, an antagonist action at the estrogen feedback receptor in the hypothalamus, an agonist action at the mammary gland estrogen receptor or an effect on adrenergic neurons in the hypothalamic-pituitary pathway. None of these has been excluded as a target because there has been a lack of a rigorous attempt to address the mechanism of action for mammary tumors at the molecular level. The potential occupational exposure to atrazine was assessed during mixing, loading and application. Absorbed daily dosage values were 1.8-6.1 microg kg(-1) day(-1). The MOS values (animal NOEL/human exposure) for short-term (acute) exposure were 820-2800. Longer-term occupational exposure and risk were also calculated. Detectable crop residues are generally absent at harvest. Theoretical calculations of acute dietary exposure used tolerance levels, along with secondary residues, and water, for which there is a maximum contamination level; atrazine plus the three main chlorotriazine metabolites were combined. MOS values were above 2000 for all population subgroups. Dietary exposure to atrazine is therefore extremely unlikely to result in human health hazard. Recent publications have reported a possible feminization of frogs, measured in laboratory and field studies. This is assumed to be due to the induction of aromatase, but no measurements of enzyme activity have been reported. In field studies, the water bodies with the greatest numbers of deformed frogs sometimes had the lowest concentrations of atrazine. Other studies have also cast doubt on the feminization theory, except perhaps at very high levels of atrazine. Epidemiology studies have investigated the possibility that atrazine may result in adverse effects in humans. Although some studies have claimed that atrazine exposure results in an elevated risk of prostate cancer, the published literature is inconclusive with respect to cancer incidence.

Animals↗

Enhanced degradation of atrazine under field conditions correlates with a loss of weed control in the glasshouse.

Enhanced degradation of atrazine has been reported in the literature, indicating the potential for reduced residual weed control with this herbicide. Experiments were conducted to determine the field dissipation of atrazine in three cropping systems: continuous Zea mays L. (CC) receiving atrazine applications each year, Gossypium hirsutum L.-Z. mays rotation (CCR) receiving applications of atrazine once every 2 years and a no atrazine history soil (NAH). Subsequent laboratory and greenhouse experiments were conducted with soil collected from these cropping systems to determine atrazine degradation, mineralization and residual weed control. Field dissipation of atrazine followed first-order kinetics, and calculated half-life values for atrazine combined over 2003 and 2005 increased in the order of CC (9 d) = CCR (10 d) < NAH (17 d). Greenhouse studies confirmed that the persistence of atrazine was approximately twofold greater in NAH soil than in CC or CCR soil. Biometer flask mineralization studies suggested that enhanced degradation of atrazine was due to rapid catabolism of the s-triazine ring. Glasshouse efficacy studies revealed a loss of residual weed control in CC and CCR soil compared with NAH soil. These data indicate that, under typical Mississippi Delta field conditions and agronomic practices, the persistence of atrazine may be reduced by at least 50% if the herbicide is applied more than once every 24 months. Glasshouse studies suggest that under these conditions a loss of residual weed control is possible.

Agriculture↗

Interaction between cadmium and atrazine during uptake by rice seedlings (Oryza sativa L.).

The uptake of atrazine by rice seedlings (Oryza sativa L.) through plant roots from nutrient solution was investigated in the presence and absence of Cd2+ over an exposure period of four weeks. It was found that both atrazine and Cd2+ were toxic to rice seedlings. Both shoot and root biomasses decreased when the seedlings were exposed to increasing atrazine or Cd2+ concentrations in nutrient solutions. In the absence of Cd2+, a linear relationship was observed between atrazine concentrations in roots/shoots and in external solution, and more atrazine is concentrated in roots than in shoots. When atrazine and Cd2+ concentrations in solution were maintained at mole ratio of 1:1, the accumulation of atrazine by seedlings was less and the seedling biomass was greater than found with other ratios, such as 1:2 or 2:1. Therefore, the formation of the complex between atrazine and Cd2+ reduced the individual toxicities. Analyses of data with the quasi-equilibrium partition model indicated that the atrazine concentrations in rice seedlings and external water were close to equilibrium. In the presence of Cd2+, however, the measured bioconcentration factor (BCF) of atrazine with roots and shoots were considerably greater. The latter findings resulted presumably from the atrazine-Cd2+ complex formation that led to a large apparent BCF.

Atrazine↗

Atrazine, deethylatrazine, and deisopropylatrazine in surface runoff from conservation tilled watersheds.

Atrazine and two of its metabolites, deethylatrazine (DEA) and deisopropylatrazine (DIA), are frequently detected in surface runoff. Although their health and environmental effects may be similar to that of atrazine and ratios of their concentrations are useful in delineating sources of contamination, there have been few long-term studies of the factors affecting their losses in runoff. Therefore, losses of atrazine, DEA, and DIA were monitored for six years in runoff from seven small (0.45-0.79 ha) watersheds under three tillage practices. Weather year and the timing of runoff-producing rainfall had a greater effect on atrazine, DEA, and DIA concentrations and losses than did tillage practice. DEA was the most frequently detected metabolite with an average concentration in the year of atrazine application, of 2.5 microg L(-1) compared to 0.7 microg L(-1) for DIA. Atrazine exceeded its 3 /g L(-1) maximum contaminant level (MCL) up to 100 days after application. DEA and DIA exceeded the atrazine MCL up to 50 days after atrazine application; thus, failure to monitor their presence may result in underestimation of the environmental impact of atrazine usage. The molar concentration ratio of DEA to atrazine (DAR) was affected by tillage treatment, weather year, and possibly soil type. These factors may need to be taken into account when DAR is used as an index of atrazine movement. The ratio of DIA to DEA (D2R) was fairly constant and should be useful in determining the parent compounds contributing DIA to surface waters.

Atrazine↗

Atrazine disrupts the hypothalamic control of pituitary-ovarian function.

The chloro-S-triazine herbicides (i.e., atrazine, simazine, cyanazine) constitute the largest group of herbicides sold in the United States. Despite their extensive usage, relatively little is known about the possible human-health effects and mechanism(s) of action of these compounds. Previous studies in our laboratory have shown that the chlorotriazines disrupt the hormonal control of ovarian cycles. Results from these studies led us to hypothesize that these herbicides disrupt endocrine function primarily through their action on the central nervous system. To evaluate this hypothesis, we examined the estrogen-induced surges of luteinizing hormone (LH) and prolactin in ovariectomized Sprague-Dawley (SD) and Long-Evans hooded (LE) rats treated with atrazine (50-300 mg/kg/day, by gavage) for 1, 3, or 21 days. One dose of atrazine (300 mg/kg) suppressed the LH and prolactin surge in ovariectomized LE, but not SD female rats. Atrazine (300 mg/kg) administered to intact LE females on the day of vaginal proestrus was without effect on ovulation but did induce a pseudopregnancy in 7 of 9 females. Three daily doses of atrazine suppressed the estrogen-induced LH and prolactin surges in ovariectomized LE females in a dose-dependent manner, but this same treatment was without effect on serum LH and prolactin in SD females. The estrogen-induced surges of both pituitary hormones were suppressed by atrazine (75-300 mg/kg/day) in a dose-dependent manner in females of both strains evaluated after 21 days of treatment. Three experiments were then performed to determine whether the brain, pituitary, or both organs were the target sites for the chlorotriazines. These included examination of the ability of (1) the pituitary lactotrophs to secrete prolactin, using hypophyosectomized females bearing pituitary autotransplants (ectopic pituitaries); (2) the synthetic gonadotropin-releasing hormone (GnRH) to induce LH secretion in females treated with high concentrations of atrazine for 3 days; and (3) atrazine (administered in vivo or in vitro) to suppress LH and prolactin secretion from pituitaries, using a flow-through perifusion procedure. In conclusion, the results of these studies demonstrate that atrazine alters LH and prolactin serum levels in the LE and SD female rats by altering the hypothalamic control of these hormones. In this regard, the LE female appeared to be more sensitive to the hormone suppressive effects of atrazine, as indicated by the decreases observed on treatment-day 3. These experiments support the hypothesis that the effect of atrazine on LH and prolactin secretion is mediated via a hypothalamic site of action.

Animals↗

Degradation and mineralization of atrazine by a soil bacterial isolate.

An atrazine-degrading bacterial culture was isolated from an agricultural soil previously impacted by herbicide spills. The organism was capable of using atrazine under aerobic conditions as the sole source of C and N. Cyanuric acid could replace atrazine as the sole source of N, indicating that the organism was capable of ring cleavage. Ring cleavage was confirmed in 14CO2 evolution experiments with [U-14C-ring]atrazine. Between 40 and 50% of ring-14C was mineralized to 14CO2. [14C]biuret and [14C]urea were detected in spent culture media. Cellular assimilation of 14C was negligible, in keeping with the fully oxidized valence of the ring carbon. Chloride release was stoichiometric. The formation of ammonium during atrazine degradation was below the stoichiometric amount, suggesting a deficit due to cellular assimilation and metabolite-N accumulation. With excess glucose and with atrazine as the sole N source, free ammonium was not detected, suggesting assimilation into biomass. The organism degraded atrazine anaerobically in media which contained (i) atrazine only, (ii) atrazine and glucose, and (iii) atrazine, glucose, and nitrate. To date, this is the first report of a pure bacterial isolate with the ability to cleave the s-triazine ring structure of atrazine. It was also concluded that this bacterium was capable of dealkylation, dechlorination, and deamination in addition to ring cleavage.

Atrazine↗

Cloning, characterization, and expression of a gene region from Pseudomonas sp. strain ADP involved in the dechlorination of atrazine.

We previously identified a Pseudomonas sp. strain, ADP, which rapidly metabolized atrazine in liquid culture, agar plates, and soils (R. T. Mandelbaum, D. L. Allan, L. P. Wackett, Appl. Environ. Microbiol. 61:1451-1457, 1995). In this study, we report the cloning and partial characterization of a gene region from Pseudomonas sp. strain ADP that encodes atrazine degradation activity. A 22-kb EcoRI genomic DNA fragment, designated pMD1, was shown to encode atrazine dechlorination activity in Escherichia coli DH5 alpha. Atrazine degradation was demonstrated by a zone-clearing assay on agar medium containing crystalline atrazine and by chromatographic methods. A gene conferring the atrazine-clearing phenotype was subsequently subcloned as a 1.9-kb AvaI fragment in pACYC184, designated pMD4, and was expressed in E. coli. This result and random Tn5 mutagenesis established that the 1.9-kb AvaI fragment was essential for atrazine dechlorination. High-pressure liquid and thin-layer chromatographic analyses were used to rigorously establish that E. coli containing pMD4 degraded atrazine and accumulated hydroxyatrazine. Hydroxyatrazine was detected only transiently in E. coli containing pMD1. This is consistent with the idea that hydroxyatrazine is the first metabolite in atrazine degradation by Pseudomonas sp. strain ADP. A 0.6-kb ApaI-PstI fragment from pMD4, containing the putative atrazine chlorohydrolase gene, hybridized to DNA from atrazine-degrading bacteria isolated in Switzerland and Louisiana.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrazine↗

Biodegradation of atrazine by Agrobacterium radiobacter J14a and use of this strain in bioremediation of contaminated soil.

We examined the ability of a soil bacterium, Agrobacterium radiobacter J14a, to degrade the herbicide atrazine under a variety of cultural conditions, and we used this bacterium to increase the biodegradation of atrazine in soils from agricultural chemical distribution sites. J14a cells grown in nitrogen-free medium with citrate and sucrose as carbon sources mineralized 94% of 50 microgram of [14C-U-ring]atrazine ml-1 in 72 h with a concurrent increase in the population size from 7.9 x 10(5) to 5.0 x 10(7) cells ml-1. Under these conditions cells mineralized the [ethyl-14C]atrazine and incorporated approximately 30% of the 14C into the J14a biomass. Cells grown in medium without additional carbon and nitrogen sources degraded atrazine, but the cell numbers did not increase. Metabolites produced by J14a during atrazine degradation include hydroxyatrazine, deethylatrazine, and deethyl-hydroxyatrazine. The addition of 10(5) J14a cells g-1 into soil with a low indigenous population of atrazine degraders treated with 50 and 200 microgram of atrazine g-1 soil resulted in two to five times higher mineralization than in the noninoculated soil. Sucrose addition did not result in significantly faster mineralization rates or shorten degradation lag times. However, J14a introduction (10(5) cells g-1) into another soil with a larger indigenous atrazine-mineralizing population reduced the atrazine degradation lag times below those in noninoculated treatments but did not generally increase total atrazine mineralization.

Atrazine↗