Bioinformatics analysis of the potential mechanisms of Alzheimer's disease induced by exposure to combined triazine herbicides.

BACKGROUND: The development of Alzheimer's disease (AD) is promoted by a combination of genetic and environmental factors. Notably, combined exposure to triazine herbicides atrazine (ATR), simazine (SIM), and propazine (PRO) may promote the development of AD, but the mechanism is unknown. AIM: To study the molecular mechanism of AD induced by triazine herbicides. METHODS: Differentially expressed genes (DEGs) of AD patients and controls were identified. The intersectional targets of ATR, SIM, and PRO for possible associations with AD were screened through network pharmacology and used for gene ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis. The binding potentials between the core targets and herbicides were validated by molecular docking and molecular dynamics. RESULTS: A total of 1,062 DEGs were screened between the AD patients and controls, which identified 148 intersectional targets of herbicides causing AD that were screened by network pharmacology analysis. GO and KEGG enrichment analysis revealed that cell cycling and cellular senescence were important signalling pathways. Finally, the core targets EGFR, FN1, and TYMS were screened and validated by molecular docking and molecular dynamics. CONCLUSION: Our results suggest that combined exposure to triazine herbicides might promote the development of AD, thereby providing new insights for the prevention of AD.

Responses to herbicides of Arctic and temperate microalgae grown under different light intensities.

In aquatic ecosystems, microalgae are exposed to light fluctuations at different frequencies due to daily and seasonal changes. Although concentrations of herbicides are lower in Arctic than in temperate regions, atrazine and simazine, are increasingly found in northern aquatic systems because of long-distance aerial dispersal of widespread applications in the south and antifouling biocides used on ships. The toxic effects of atrazine on temperate microalgae are well documented, but very little is known about their effects on Arctic marine microalgae in relation to their temperate counterparts after light adaptation to variable light intensities. We therefore investigated the impacts of atrazine and simazine on photosynthetic activity, PSII energy fluxes, pigment content, photoprotective ability (NPQ), and reactive oxygen species (ROS) content under three light intensities. The goal was to better understand differences in physiological responses to light fluctuations between Arctic and temperate microalgae and to determine how these different characteristics affect their responses to herbicides. The Arctic diatom Chaetoceros showed stronger light adaptation capacity than the Arctic green algae Micromonas. Atrazine and simazine inhibited the growth and photosynthetic electron transport, affected the pigment content, and disturbed the energy balance between light absorption and utilization. As a result, during high light adaptation and in the presence of herbicides, photoprotective pigments were synthesized and NPQ was highly activated. Nevertheless, these protective responses were insufficient to prevent oxidative damage caused by herbicides in both species from both regions, but at different extent depending on the species. Our study demonstrates that light is important in regulating herbicide toxicity in both Arctic and temperate microalgal strains. Moreover, eco-physiological differences in light responses are likely to support changes in the algal community, especially as the Arctic ocean becomes more polluted and bright with continued human impacts.

Consequences of contamination on the interactions between phytoplankton and bacterioplankton.

Sediment resuspension can provoke strong water enrichment in nutrients, contaminants, and microorganisms. Microcosm incubations were performed in triplicate for 96 h, with lagoon and offshore waters incubated either with sediment elutriate or with an artificial mixture of contaminants issued from sediment resuspension. Sediment elutriate provoked a strong increase in microbial biomass, with little effects on the phytoplankton and bacterioplankton community structures. Among the pool of contaminants released, few were clearly identified as structuring factors of phytoplankton and bacterioplankton communities, namely simazine, Cu, Sn, Ni, and Cr. Effects were more pronounced in the offshore waters, suggesting a relative tolerance of the lagoon microbial communities to contamination. The impacts of contamination on the microbial community structure were direct or indirect, depending on the nature and the strength of the interactions between phytoplankton and bacterioplankton.

Solid lipid nanoparticles co-loaded with simazine and atrazine: preparation, characterization, and evaluation of herbicidal activity.

Solid lipid nanoparticles (SLN) containing the herbicides atrazine and simazine were prepared and characterized, and in vitro evaluation was made of the release kinetics, herbicidal activity, and cytotoxicity. The stability of the nanoparticles was investigated over a period of 120 days, via analyses of particle size, ζ potential, polydispersion, pH, and encapsulation efficiency. SLN showed good physicochemical stability and high encapsulation efficiencies. Release kinetics tests showed that use of SLN modified the release profiles of the herbicides in water. Herbicidal activity assays performed with pre- and postemergence treatment of the target species Raphanus raphanistrum showed the effectiveness of the formulations of nanoparticles containing herbicides. Assays with nontarget organisms (Zea mays) showed that the formulations did not affect plant growth. The results of cytotoxicity assays indicated that the presence of SLN acted to reduce the toxicity of the herbicides. The new nanoparticle formulations enable the use of smaller quantities of herbicide and therefore offer a more environmentally friendly method of controlling weeds in agriculture.

Toxicity endpoint selections for a simazine risk assessment.

BACKGROUND: California uses simazine at one of the highest levels for states in the United States (approximately 2.5 million lbs 2006-2010). Simazine causes neuroendocrine disruption and mammary cancer in test animals. A risk assessment was prioritized by the California Department of Pesticide Regulation because of the nondietary concern for simazine exposure to occupational/nonoccupational simazine users, resident nonusers, and bystanders (especially children and children exhibiting pica) at greatest risk. METHODS: No observed effect levels (NOELs) from animal studies as well as human exposure data were used to determine nondietary values for the above populations. Registrant-submitted and open literature studies focusing on oral (major human route) effects for simazine and the major metabolites desisopropyl-s-atrazine and diaminochlorotriazine were reviewed as part of the hazard identification process. RESULTS: Developmental, reproduction, and chronic studies provided the lowest NOELs for the acute (5 mg/kg/day), subchronic (0.56 mg/kg/day), and chronic (0.52 mg/kg/day) exposure durations, respectively. A benchmark dose (95th percentile) was calculated for mammary tumorigenesis, assuming a threshold mechanism in rats (benchmark dose lower limit [95th percentile; BMDL05 ]: 2.9 mg/kg/day). Margins of exposure and uncertainty factors (100-300×, depending on exposure scenario) were used to characterize risk for designated population subgroups. CONCLUSIONS: Fetal developmental delays, endocrine disruption, and mammary tumors resulted from simazine treatment. Systemic and maternal/fetal effects determined the critical NOELs used in risk assessment. Margins of exposures for most scenarios were below acceptable levels, especially for children who may be bystanders where simazine is applied and children who exhibit pica. This risk characterization raises a concern for long-term effects in humans.

Effects of subchronic exposure to simazine on zebrafish (Danio rerio).

OBJECTIVES: Simazine is a triazine herbicide which has been used for a long period in agriculture and in the aquatic environment for control of weeds and algae. The aim of this study was to investigate the effects of subchronic exposure to simazine on growth and the development of histopathological changes in selected organs (gills, kidney, liver) in Danio rerio. METHODS: Juvenile growth tests were performed on D. rerio according to the OECD method No. 215. Fish at the age of 20 days were exposed to the environmental simazine concentration commonly detected in Czech rivers (0.06 µg.L-1) and a range of sublethal concentrations of simazine (0.6, 6.0 and 60.0 µg.L-1) for 28 days. RESULTS: There were no significant differences (p<0.05) between the specific growth rates (r) of the test groups and those of both control groups. Histopathological examination revealed pathological changes in fish exposed to a simazine concentration of 60.0 µg.L-1. The values of NOEC and LOEC of simazine were 6.0 µg.L-1 and 60.0 µg.L-1. CONCLUSIONS: The environmental concentration of simazine in Czech rivers did not have any effects on the growth and development of histopathological changes in D. rerio.

Simazine application inhibits nitrification and changes the ammonia-oxidizing bacterial communities in a fertilized agricultural soil.

s-Triazine herbicides are widely used for weed control, and are persistent in soils. Nitrification is an essential process in the global nitrogen cycle in soil, and involves ammonia-oxidizing Bacteria (AOB) and ammonia-oxidizing Archaea (AOA). In this study, we evaluated the effect of the s-triazine herbicide simazine on the nitrification and on the structure of ammonia-oxidizing microbial communities in a fertilized agricultural soil. The effect of simazine on AOB and AOA were studied by PCR-amplification of amoA genes of nitrifying Bacteria and Archaea in soil microcosms and denaturing gradient gel electrophoresis (DGGE) analyses. Simazine [50 µg g(-1) dry weight soil (d.w.s)] completely inhibited the nitrification processes in the fertilized agricultural soil. The inhibition by simazine of ammonia oxidation observed was similar to the reduction of ammonia oxidation by the nitrification inhibitor acetylene. The application of simazine-affected AOB community DGGE patterns in the agricultural soil amended with ammonium, whereas no significant changes in the AOA community were observed. The DGGE analyses strongly suggest that simazine inhibited Nitrosobacteria and specifically Nitrosospira species. In conclusion, our results suggest that the s-triazine herbicide not only inhibits the target susceptible plants but also inhibits the ammonia oxidation and the AOB in fertilized soils.

Microalgae fiber optic biosensors for herbicide monitoring using sol-gel technology.

Three microalgal species (Dictyosphaerium chlorelloides (D.c.), Scenedesmus intermedius (S.i.) and Scenedesmus sp. (S.s.)) were encapsulated in silicate sol-gel matrices and the increase in the amount of chlorophyll fluorescence signal was used to quantify simazine. Influence of several parameters on the preparation of the sensing layers has been evaluated: effect of pH on sol-gel gelation time; effect of algae density on sensor response; influence of glycerol (%) on the membrane stability. Long term stability was also tested and the fluorescence signal from biosensors remained stable for at least 3 weeks. D.c. biosensor presented the lowest detection limits for simazine (3.6 microg L(-1)) and the broadest dynamic calibration range (19-860 microg L(-1)) with IC(50) 125+/-14 microg L(-1). Biosensor was validated by HPLC with UV/DAD detection. The biosensor showed response to those herbicides that inhibit the photosynthesis at photosystem II (triazines: simazine, atrazine, propazine, terbuthylazine; urea based herbicides: linuron). However, no significant increases of fluorescence response was obtained for similar concentrations of 2,4-D (hormonal herbicide) or Cu(II). The combined use of two biosensors that use two different genotypes, sensitive and resistant to simazine, jointly allowed improving microalgae biosensor specificity.

Effect of long-term exposure to simazine on histopathology, hematological, and biochemical parameters in Cyprinus carpio.

We investigated if residues of simazine in the natural waters would cause histological, hematological, and biochemical alterations in carps from contaminated areas in Badajoz (Spain). Some necrotic foci in kidney and liver, hepatitis, and hepatic steatosis were detected. No changes on measured hematological and biochemical parameters between fish from reference and contaminated ponds were observed. To assess if simazine exposure was the cause of these observations carps were exposed in the laboratory to simazine (45 microg/L) for 90 days. Some results obtained in the field were confirmed in laboratory, such as necrosis in kidney and liver and hepatic steatosis. Globular eosinophilic foci in kidney and a slight decrease of the hematocrit were also detected. These changes were moderate and indicative of an adaptation of the fish to the toxic stress caused by exposure to low simazine concentrations.

Evidence for cross-adaptation between s-triazine herbicides resulting in reduced efficacy under field conditions.

BACKGROUND: Enhanced atrazine degradation has been observed in agricultural soils from around the globe. Soils exhibiting enhanced atrazine degradation may be cross-adapted with other s-triazine herbicides, thereby reducing their control of sensitive weed species. The aims of this study were (1) to determine the field persistence of simazine in atrazine-adapted and non-adapted soils, (2) to compare mineralization of ring-labeled (14)C-simazine and (14)C-atrazine between atrazine-adapted and non-adapted soils and (3) to evaluate prickly sida control with simazine in atrazine-adapted and non-adapted soils. RESULTS: Pooled over two pre-emergent (PRE) application dates, simazine field persistence was 1.4-fold lower in atrazine-adapted than in non-adapted soils. For both simazine and atrazine, the mineralization lag phase was 4.3-fold shorter and the mineralization rate constant was 3.5-fold higher in atrazine-adapted than in non-adapted soils. Collectively, the persistence and mineralization data confirm cross-adaptation between these s-triazine herbicides. In non-adapted soils, simazine PRE at the 15 March and 17 April planting dates reduced prickly sida density at least 5.4-fold compared with the no simazine PRE treatment. Conversely, in atrazine-adapted soils, prickly sida densities were not statistically different between simazine PRE and no simazine PRE at either planting date, thereby indicating reduced simazine efficacy in atrazine-adapted soils. CONCLUSIONS: Results demonstrate the potential for cross-adaptation among s-triazine herbicides and the subsequent reduction in the control of otherwise sensitive weed species.

Transgenic rice plants expressing human CYP1A1 remediate the triazine herbicides atrazine and simazine.

The human cytochrome P450 CYP1A1 gene was introduced into rice plants (Oryza sativa cv. Nipponbare). One-month-old CYP1A1 plants grown in soil clearly showed a healthy growth and tolerance to 8.8 microM atrazine and 50 microM simazine, but nontransgenic plants were completely killed by the herbicides. Although transgenic and nontransgenic plants metabolized the two herbicides into the same sets of compounds, CYP1A1 plants metabolized atrazine and simazine more rapidly than did control plants. In small-scale experiments, residual amounts of atrazine and simazine in the culture medium of CYP1A1 plants were 43.4 and 12.3% of those in control medium; those of nontransgenic Nipponbare were 68.3 and 57.2%, respectively. When cultivated in soil with 2.95 microM atrazine and 3.15 microM simazine for 25 days, CYP1A1 plants eliminated 1.3 times more atrazine and 1.4 times more simazine from the soil than did control plants. Thus, CYP1A1 rice plants make it possible to remove atrazine and simazine more rapidly from the culture medium and soil than can nontransgenic Nipponbare.

[Anti-corrosive effect of pesticides in soil corrosion conditions]. / Antikorrozionnoe deistvie nekotorykh pestitsidov v usloviiakh pochvennoi korrozii.

Effect of some sub-standard pesticides (Ramrod, Linuron, Simazin) with respect to corrosion-active groups of microorganisms: sulphate-reducing bacteria (SRB), denitrifying bacteria (DNB), saprophytic bacteria (SB) and their inhibiting properties under the conditions of active corrosion have been studied to estimate a possibility to use them as biocide additions when producing protective materials. It has been shown that the sub-standard pesticides Ramrod and Simazin are promising for to be used as the biocides additions under the protection of bioresistant materials. It is supposed that inhibitors-biocides may be found in a series of compounds obtained under chemical modification of substandard pesticides Ramrod and Simazin.

The impact of two pesticides on olfactory-mediated endocrine function in mature male Atlantic salmon (Salmo salar L.) parr.

Short-term exposure of the olfactory epithelium of mature male Atlantic salmon parr to either the pesticide simazine (concentrations 1.0 and 2.0 microg l(-1)) or the pesticide atrazine (concentration 1.0 microg l(-1)) significantly reduced the olfactory response to the female priming pheromone, prostaglandin F(2alpha). In addition, the reproductive priming effect of the pheromone on the levels of expressible milt was also reduced after exposure to the individual pesticides (simazine 0.1, 0.5, 1.0 and 2.0 microg l(-1) and atrazine 0.5 and 2.0 microg l(-1)). When the olfactory epithelium was exposed to a mixture of simazine and atrazine, (concentrations of 0.5:0.5 and 1.0:1.0 microg l(-1)), there was no significant reduction in the olfactory response when compared to the single pesticides at equivalent concentrations. In addition, exposure to a mixture of simazine and atrazine had no synergistic effect on the priming response, and plasma levels of testosterone, 11-ketotestosterone and 17,20beta-dihydroxy-4-pregnen-3-one were similar in the groups of male parr exposed to the individual pesticides. Although the levels of expressible milt were reduced in all groups, there were no significant differences between the different pesticide treatments. The results of the study suggest that the two s-triazine pesticides have an additive and not a synergistic impact on olfactory-mediated endocrine function in mature male salmon parr.

Determination of resistant biotypes of Amaranthus retroflexus L. on triazines.

Considering the fact that since 1966 in our country the mostly produced have been triazine herbicides on the bases of ametryn, simazine, atrazine and prometryn, we have studied resistance of Amaranthus retroflexus from different sites in regard to the above mentioned herbicides. Seed of weed species for which exist-possibility of resistance have been collected from different localities in Vojvodina, such as Backa Palanka, Backi Maglic and Becej. Studies were performed during 1999 and 2000 by whole plant studies (Thurnwachter, 1998) and by petri dish assays (Clay, Underwood, 1989). Plants were treated by range of atrazine rates (Atrazin S-50) including also susceptible, referent population. Results indicate atrazine resistance of Amaranthus retroflexus at Backa Palanka and Backi Maglic sites, which have been treated by triazine herbicides in many years lasting period.

4-Sulfenyl-2-carbamoyl-4-isoxazolin-3-ones: biological isostere to 4-chloro-2-carbamoyl-4-isoxazolin-3-ones.

4-Sulfenyl-2-carbamoyl-4-isoxazolin-3-ones (4) were designed on the basis of biological isosterism and prepared in four steps. Some of these compounds showed sufficient pre-emergent herbicidal activities against various kinds of weeds. Among the synthesized compounds, 2-(N-(4-chlorophenyl)-N-isopropylcarbamoyl)-4-ethylthio-5-methyl-4 -isoxazolin-3-one (4cd) exhibited the most promising activity.

Changes in rat liver cytochrome P450 enzymes by atrazine and simazine treatment.

1. We examined the effect of two chloro-s-triazines (atrazine and simazine) on hepatic microsomal cytochrome P450 enzymes in rat. Rats were treated intraperitoneally with atrazine or simazine daily for 3 days with 100, 200 and 400 mumol/kg. 2. Among the P450-dependent monooxygenase activities, testosterone 2 alpha-hydroxylase (T2AH) activity in rat, which is associated with CYP2C11, was significantly decreased at all doses of atrazine and simazine. The levels relative to control activities were 59-46 and 60-32% respectively. Similarly, oestradiol 2-hydroxylase (ED2H) activity was also significantly decreased by 28-51% by atrazine and simazine at all doses. However, no change in CYP2C11 protein level by either chloro-s-triazine was observed. K(m) for T2AH was significantly increased only by simazine (200 mumol/kg), whereas the Vmax and Cl(int) for T2AH were significantly decreased by atrazine and simazine at all doses. 3. 7-Ethoxyresorufin O-deethylase (EROD), 7-methoxyresorufin O-demethylase (MROD) and 7-pentoxyresorufin O-depentylase (PROD) activities were significantly increased by 1.4-1.6-, 1.7-3.2- and 1.5-2.2-fold respectively, by both chloro-s-triazines at 200 or 400 mumol/kg. Lauric acid omega-hydroxylase (LAOH) was also increased by 1.4-fold by simazine at 200 and 400 mumol/kg. Immunoblotting showed that only simazine induces CYP1A2 and CYP4A1/2 protein expression. 4. The activities of 7-ethoxycoumarin O-deethylase (ECOD), bufuralol 1'-hydroxylase (BF1'H), chlorzoxazone 6-hydroxylase (CZ6H), testosterone 6 beta-hydroxylase (T6BH) and testosterone 7 alpha-hydroxylase (T7AH) were not affected by either chloro-s-triazine. 5. These results suggest that the pattern of changes in P450 isoforms by chloro-s-triazines differs between atrazine and simazine, that these herbicides change the constitutive and/or male specific P450 isoform(s) in rat liver, and that these changes closely relate to the toxicity of chloro-s-triazines.

The inhibition of estrogen receptor-mediated responses by chloro-S-triazine-derived compounds is dependent on estradiol concentration in yeast.

The chloro-S-triazine derived compounds atrazine, atrazine desisopropyl, cyanazine, and simazine are commonly used herbicides. These compounds do not have estrogenic activity in yeast expressing human estrogen receptor (hER) and an estrogen-sensitive reporter. In the presence of a concentration of estradiol (20 nM) that induced maximal reporter activity in yeast, the triazines did not inhibit reporter activity. However, the triazines decreased reporter activity in a dose dependent manner in the presence of a submaximal concentration of estradiol (0.5 nM). The estradiol-dependent activity of a mutant hER lacking the amino terminus was not inhibited by the triazines in yeast. Competition binding assays demonstrated that the triazines displaced radiolabeled estradiol from recombinant hER. These results suggest that the ability of the triazines to inhibit estrogen receptor-mediated responses in yeast occur through their interaction with hER and is dependent on the concentration of estradiol.

Failure of chloro-S-triazine-derived compounds to induce estrogen receptor-mediated responses in vivo and in vitro.

The potential estrogenic activities of atrazine and simazine were investigated in vivo using the immature female Sprague-Dawley rat uterus and in vitro using the estrogen-responsive MCF-7 human breast cancer cell line and the estrogen-dependent recombinant yeast strain PL3. Animals that were dosed with 50, 150, or 300 mg/kg of atrazine or simazine alone for 3 consecutive days did not exhibit any significant increases in uterine wet weight while decreases in cytosolic progesterone receptor (PR) binding levels and uterine peroxidase activity were observed. 17 beta-estradiol (E2)-induced increases in uterine wet weight were not significantly affected by cotreatment with either chemical; however, some dose-independent decreases in E2-induced cytosolic PR binding and uterine peroxidase activity were observed. In vitro, atrazine and simazine did not affect basal or E2-induced MCF-7 cell proliferation or the formation of nuclear PR-DNA complexes as determined by gel electrophoretic mobility shift assays. In addition, these chloro-S-triazines did not display agonist activity or antagonize E2-induced luciferase activity in MCF-7 cells transiently transfected with a Gal4-human estrogen receptor chimera (Gal4-HEGO) and a Gal4-regulated luciferase reporter gene (17m5-G-Luc). Moreover, the estrogen-dependent PL3 yeast strain was not capable of growth on minimal media supplemented with atrazine or simazine in place of E2. Collectively, these results indicate that the reported estrogenic and antiestrogenic effects elicited by these chemicals are not mediated by the estrogen receptor.

Characterization of the expression of the thcB gene, coding for a pesticide-degrading cytochrome P-450 in Rhodococcus strains.

A cytochrome P-450 system in Rhodococcus strains, encoded by thcB, thcC, and thcD, participates in the degradation of thiocarbamates and several other pesticides. The regulation of the system was investigated by fusing a truncated lacZ in frame to thcB, the structural gene for the cytochrome P-450 monooxygenase. Analysis of the thcB-lacZ fusion showed that the expression of thcB was 10-fold higher in the presence of the herbicide EPTC (s-ethyl dipropylthiocarbamate). Similar enhancement of the thcB-lacZ expression was found with other thiocarbamate pesticides. Atrazine, simazine, or carbofuran, although metabolized by the system, had no effect on the thcB-lacZ expression. The presence of glucose slightly increased the expression of thcB-lacZ, indicating no catabolic repression of the thcB-lacZ expression. The expression of thcB-lacZ was decreased more than twofold in Luria-Bertani medium. This was due in part to cysteine, which repressed thcB-lacZ expression. It was confirmed that the thcR gene, which is transcribed divergently from thcB, codes for a positive regulatory protein which is essential for the thcB-lacZ expression. Studies of the thcR-lacZ protein fusion showed that the thcR gene is expressed constitutively.

Short-term effects of chlorotriazines on estrus in female Sprague-Dawley and Fischer 344 rats.

Atrazine or simazine (s-chlorotriazines) was administered by gavage daily for 2 wk to female Sprague-Dawley and Fischer 344 rats at oral doses of 100 or 300 mg/kg to evaluate effects on body, ovary, uterus, and adrenal weights, estrous cycle stages, vaginal cytology, and plasma hormone (estradiol, progesterone, prolactin, and corticosterone) levels. Significant reductions in body weights of both Sprague-Dawley and Fischer 344 female rats at both dose levels were accompanied by a significant reduction in ovarian and uterine weights, and a decrease in circulating estradiol levels. The magnitudes of the effects were less in Fischer 344 rats than in Sprague-Dawley rats, and the effects of simazine were less pronounced than those of atrazine at the same dose. A maximum tolerated dose (MTD: > or = 10% body weight reduction) was estimated to be 100 mg/kg for atrazine and 300 mg/kg for simazine for both stains. The Sprague-Dawley female rats exhibited a treatment-related lengthening of the estrous cycle and an increased number of days characterized by cornified epithelial cells. This resulted in a greater percent of the cycle days spent in estrus and reduction in the percent of the cycle days spent in diestrus. Atrazine-dosed Fischer 344 females also exhibited a significant trend toward cycle lengthening, but this was due to reduction in the percent of cycle spent in estrus and a concomitant increase in diestrual days. These findings suggest that treatment with doses of triazine at or above the MTD may result in prolonged exposure to endogenous estrogen in the Sprague-Dawley but not the Fischer 344 rat. These changes may account for the observed earlier onset and/or increased incidence of mammary tumors in chlorotriazine-treated female Sprague-Dawley rats. This strain of rat is already known to be prone to a substantial development of mammary tumors with advancing age, while the Fischer 344 strain is not as likely to exhibit this response.