Page 1 of 14 

Atrazine Activates an ACTH-Dependent Release of Corticosterone and 

Progresterone in Male Wistar Rats Following Acute Exposure 

Internal Report Prepared for the use of the Office of Pesticide Program, 

Office of Chemical Safety and Pollution Prevention,

U.S. Environmental Protection Agency, 

Washington, DC 

July 15, 2010 

U.S. EPA/ORD Contributors 

Michelle Gatien Hotchkiss, Deborah S. Best, Ralph L. Cooper, Susan C. Laws 

Endocrine Toxicology Branch, Toxicity Assessment Division, 

National Health and Environmental Effects Research Laboratory, 

Office of Research and Development, 

U.S. Environmental Protection Agency, 

 Research Triangle Park, North Carolina 27711 

This document has been reviewed in accordance with the policy of the National Health and 

Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, and 

approved for publication. Approval does not signify that the contents necessarily reflect the 

views or policy of the Agency nor does mention of trade names or commercial products 

constitute endorsement or recommendation for use. 
Page 2 of 14 

Table of Contents

Executive Summary............................................................................3 

Background.......................................................................................4 

Materials and Methods..........................................................................5 

Results.............................................................................................7 

Discussion.........................................................................................8 

References.......................................................................................10 

Figure 1...........................................................................................13 

Table 1...........................................................................................14 
Page 3 of 14 

Executive Summary: 

Previous studies conducted in the laboratory of Dr. Susan C. Laws have shown that a 

single oral administration of atrazine (ATR; 50-200 mg/kg, oral gavage), a chlorotriazine 

herbicide, induces dose-dependent increases in plasma adrenocorticotropic hormone (ACTH), 

serum corticosterone (CORT) and progesterone in male Wistar rats(Toxicol Sci., 112(1):78-87). 

The clear and rapid increase in ACTH followed by an increase in the adrenal steroid hormones 

(corticosterone and progesterone) have defined a novel endocrine effect of the chlorotriazines not 

previously characterized. Additional studies in our laboratory have indicated that this chemicalinduced change in the hypothalamic-pituitary-adrenal axis (HPA-axis) activation is not due to a 

generalized stress response to animal handling or gastrointestinal distress (Laws et al., 2009; 

Fraites et al., 2009; unpublished observations in). While the cellular events leading to the 

activation of the HPA-axis are not known, extended elevation of adrenal steroids can adversely 

impact a number of homeostatic mechanisms including normal functioning of the hypothalamicpituitary-gonadal (HPG)-axes. As part of an extensive re-evaluation of the human health effects 

of atrazine, the Office of Chemical Safety and Pollution Prevention, U.S. Environmental 

Protection Agency (U.S. EPA), is reviewing studies that have characterized the effects of 

atrazine and its dealkylated metabolites on adrenal steroid synthesis. During the new evaluation, 

U.S. EPA will consider the potential for cancer and non-cancer effects of atrazine, and will 

include data generated since 2003 from laboratory and population studies. To be certain that the 

best science possible is used in its assessment of the atrazine human health risk and to ensure 

transparency, U.S. EPA is seeking advice from an independent Scientific Advisory Panel (SAP) 

established under the Federal Insecticide, Fungicide and Rodenticide Act. The data in this report 

are provided to support the third meeting of the SAP peer review with its evaluation of cancer 

and non-cancer effects of atrazine based on animal toxicology studies. 

A fundamental requirement for a better understanding of the effects of atrazine on the 

HPA-axis is the characterization of the dose-response relationship between exposure to the 

herbicide and the hormonal response. Although a partial description of acute (single dose) 

atrazine-induced changes in ACTH, corticosterone, and progesterone in the male Wistar rat was 

published by Laws et al. (2009); however a better characterization of the lower end of the doseresponse curve was needed (5-50 mg/ kg). In response to this need, the current report 
Page 4 of 14 

summarizes subsequent experiments conducted within National Health and Environmental 

Effects Research Laboratory (NHEERL) to confirm and extend the results reported in Laws et 

al., 2009. The 15 min post-dosing collection time point was chosen for these additional 

experiments based on the previous observation that elevations in plasma ACTH concentrations 

after atrazine treatment peak at approximately 15 min and begin to decline by 30 min (Laws et 

al., 2009; Figure 1). For the current study, data demonstrate that the LOEL for elevations in 

plasma concentrations of ACTH 15 min following a single dose of atrazine (oral gavage), is 12.5 

mg/kg, and 5 mg/kg for both serum corticosterone and progesterone. To evaluate the 

reproducibility and robustness of the response, the study was conducted in two separate blocks, 

with each block containing a full dose-response for atrazine (Table 1). An additional diamino-schlorotriazine (DACT) group was included to replicate the earlier observation that an equal 

molar dose of this di-dealkylated metabolite of atrazine produces an effect on the HPA-axis of 

much smaller magnitude than that produced by the parent compound. These data are in 

agreement with previously published results (Laws et al., 2009), and are provided as 

supplemental data that can be further evaluated using a benchmark analysis. 

Background 

Atrazine (ATR), a chlorotriazine herbicide, is widely used in the United States for 

broadleaf weed control in several commercial crops and landscape vegetation. 

Biotransformation by-products of ATR, as well as the parent compound, are relatively persistent 

in water and mobile in soil (Seiler et al., 1992) and have been detected in surface and ground 

water in areas of major usage, with highest levels detected most commonly in the Midwestern 

states during the growing season (Baker, 1998; Balu et al., 1998). Environmental exposure to 

ATR and its biotransformation by-products has generated much concern about the potential 

adverse effects of these chemicals, resulting in a large body of research on this subject over the 

past 10 years. Primarily, exposure to ATR and its biotransformation by-products has been 

observed to disrupt the development and function of the mammalian reproductive system by 

causing neuroendocrine effects (Cooper et al., 1996, 2000, 2007; Foradori et al.,2009a,b), delays 

in pubertal development in male and female rats (Laws et al., 2000, 2003; Stoker et al., 2000, 

2002), and altered gene expression in the steroidogenic pathway (Sanderson et al., 2000; Suzawa 

and Ingraham 2008). ATR administration has also been observed to increase serum 
Page 5 of 14 

concentrations of estradiol and estrone (Stoker et al., 2000), progesterone, corticosterone 

(CORT), and plasma adrenocorticotropic hormone (ACTH; Fraites et al., 2009; Laws et al., 

2009; Pruit et al., 2003). 

This report provides data from a study conducted to better characterize the lowest 

observed effect level (LOEL) for the effects of atrazine on plasma ACTH, corticosterone and 

progesterone concentrations in male Wistar rats 15 min post-dosing. The 15 min post-dosing 

time point was chosen for these additional experiments based on the previous observation that 

elevations in plasma ACTH concentrations after atrazine treatment peak at approximately 15 min 

and begin to decline by 30 min (Laws et al., 2009, Figure 1). To evaluate the reproducibility and 

robustness of the response, the study was conducted in two separate blocks, with each block 

containing a full dose response for atrazine (5-200 mg/kg). Data from the blocks were analyzed 

separately, and then combined and tested for block and treatment effects, and block x treatment 

interactions. An additional diamino-s-chlorotriazine (DACT, 135 mg/kg) group was included as 

a replicate of the earlier study by Laws et al. 2009. These data are submitted to the Office of 

Chemical Safety and Pollution Prevention, U.S. EPA., Washington, DC, as supplemental data for 

further review and benchmark analysis. 

Materials and Methods 

Animals 

Male Wistar rats, 60 days of age were obtained from Charles River, Inc, (Raleigh, NC) 

and were maintained under controlled temperature (20–24°C) and humidity (40–50%). To 

acclimate the animals to the dosing procedure, all rats were pre-dosed daily by oral gavage with 

1% methylcellulose vehicle for one week prior to administration of the test chemical. Previous 

experience has shown that such handling of the animals prior to actual exposure to the test 

chemical provides more consistent ACTH and corticosterone concentrations during the morning 

hours when the circadian fluctuation of these hormones is the lowest (Laws et al., 2009). 

Animals were ranked by body weight and placed into treatment groups such that the mean body 

weights + SE for all groups were similar. All experiments were approved by the U.S. EPA, 

NHEERL’s Institutional Animal Care and Use Committee (IACUC) and was conducted in 

accordance with National Institute of Health standards for laboratory animal research (LAPR# 

12-06-005). 
Page 6 of 14 

Dosing solutions and procedures. 

ATR (2-chloro-4-ethylamino-6-isopropylamino-s-triazine, CAS 1912-24-9, purity 97.1%; 

a gift from Syngenta Crop Protection, Greensboro, NC), and DACT (diamino-s-chlorotriazine, 

CAS 3397-62-4, purity 97%, ACROS Organics, Belgium; A017756301) were administered by 

oral gavage in a suspension of 1% methyl cellulose/distilled water (CAS 9004-67-5, M-7140, 

Sigma-Aldrich Chemical Company, St. Louis, MO) in a volume of 5.0 ml dosing solution/ kg 

body weight. Control animals received 1% methylcellulose only (5.0 ml dosing solution/ kg 

body weight). To facilitate the comparison of the potency of each test chemical with that of 

ATR, the dose selected for DACT is the molar equivalent of ATR 200mg/kg. Doses of ATR and 

DACT were selected based on results reported in previous studies of ATR on female pubertal 

development and HPA-axis activation (Laws et al., 2000, 2003, 2009).

Experimental Design: Characterization of the HPA-axis response to ATR 15 min after a 

single administration. 

In order to further characterize the HPA-axis response to ATR across a lower range of 

doses, a dose-response experiment was conducted to measure plasma ACTH and serum CORT, 

progesterone, and body temperature 15 min after a single administration of atrazine. Animals 

were housed under controlled lighting conditions (12 hr light/12 hr dark; lights on at 0700 EST 

and off at 1900 EST) and given ad libitum access to food (Purina Laboratory Rat Chow, 5001) 

and filtered tap water. On the day of necropsy, animals were given one dose of either vehicle 

(methyl cellulose), ATR (5, 12.5, 25, 50, 100, or 200 mg/kg), or DACT (135 mg/kg), by oral 

gavage at 0900 EST and killed by decapitation 15 min post-dosing. Animals were housed in a 

room immediately adjacent to the necropsy room to limit the transfer time of the animals from 

the home cage to the necropsy room to within 15 seconds. Upon decapitation, blood was 

processed for serum and plasma, and aliquots were stored at -80°C until measurement of ACTH, 

CORT, and progesterone by radioimmunoassay (RIA). To evaluate the reproducibility and 

robustness of the effects, the study was conducted in two experimental blocks with each 

treatment group containing ten animals/ treatment group. 
Page 7 of 14 

Radioimmunoassays: 

Total serum CORT and progesterone were measured using Coat-A-Count RIA kits from 

Siemens Medical Solutions Diagnostics (TKRC and TKPG; Los Angeles, CA). The limits of 

sensitivity for these assays were 5.7ng/ml and 0.02ng/ml, respectively. Total plasma ACTH was 

measured using an RIA kit from MP Biomedicals (07-106102, Orangeburg, NY). Crossreactivity of human ACTH antibody with rat ACTH was tested using serial dilutions of rat 

plasma with and without spikes of ACTH. Linearity of ACTH concentration was confirmed 

across all dilutions of rat plasma and in the presence of rat plasma samples spiked with ACTH. 

The limit of sensitivity for this assay was 5.7 pg/ml. 

Statistical Analysis 

Treatment means for each dose group were tested for homogeneity of variance using the 

Bartlett test (Graph Pad Instat; Graph Pad Software, San Diego, CA). As heterogeneity of 

variance was evident for each hormone tested, data were log transformed prior to further 

analysis. Data were analyzed by ANOVA using the general linear model (GLM) procedure 

(Statistical Analysis System; SAS Institute, Inc., Cary, NC) for block and treatment effects, and 

treatment x block interactions. Subsequently, since no significant block or block x treatment 

interactions were observed for any of the hormones, data from both blocks were combined for 

statistical analysis. In cases where a significant treatment effect (p< 0.05) was observed, the 

ATR dose-response data were further analyzed using Least Squared Means post-hoc tests to 

compare control values to those for each treatment group. A two-tailed Mann Whitney test 

(Graph Pad Instat; Graph Pad Software, San Diego, CA) was used to compare the DACT group 

to the control group. All post-hoc and Mann Whitney results were considered significant when p 

< 0.05. Data are reported as mean + SEM, (n=10/treatment for each block unless otherwise 

specified). 

Results

Effects of a Single Dose of ATR or DACT on ACTH, CORT, and Progesterone 

Concentrations 

ACTH, CORT, and progesterone concentrations 15 min following a single dose of ATR 

or DACT are shown in Table 1 for each experimental block. Hormone concentrations for the 
Page 8 of 14 

vehicle control animals were within range of those typically observed in untreated control 

animals in our laboratory, suggesting that the experimental design had no impact on these 

measures (Laws et al, 2009). A dose-dependent increase in plasma ACTH concentrations was 

observed following ATR administration, with significant increases observed at doses of 

12.5mg/kg and above (Table 1, NOEL= 5mg/kg). Similarly, ATR treatment resulted in dosedependent increases in serum CORT and progesterone concentrations with significant increases 

observed at all doses tested (5-200 mg/kg; Table 1, LOEL= 5mg/kg). Although a single 

administration of DACT (135 mg/ kg) caused a significant increase in ACTH (Block 1 only), 

CORT and progesterone concentrations, this increase was substantially less than that observed 

for an equal molar dose of ATR (200 mg/kg, Table 1). This observed difference between the 

potency of equal molar doses of ATR and DACT agrees with previous reports from this 

laboratory (Laws et al., 2009). 

Discussion 

The data reported here confirm previous findings in this laboratory that a single 

administration of ATR causes a significant elevation in plasma ACTH and serum CORT and 

progesterone concentrations 15 min post-administration (50-200 mg/kg; Laws et al., 2009; 

Fraites et al., 2009). The current data also extend previous findings on the effects of atrazine on 

ACTH, CORT, and progesterone by adding additional doses at the lower end of the dose 

response curve (5-50 mg/kg). Previous work (Laws et al., 2009) and the current studies 

demonstrate that a single dose of ATR produces a reproducible and robust elevation in ACTH 

(12.5-200 mg/ kg), CORT (5-200 mg/kg), and progesterone (5-200 mg/kg) concentrations that is 

both dose-and time-responsive. In all studies to date, peak elevations of ACTH concentrations 

following a single dose of ATR occur approximately 15 min before those for CORT and 

progesterone, suggesting that the observed activation of the HPA-axis is likely mediated through 

a neuroendocrine pathway (e.g., corticotrophin releasing hormone release from the 

hypothalamus). However, this does not rule out the possibility that ATR may have effects at 

multiple levels of the HPA-axis simultaneously (i.e. the hypothalamus, pituitary and/or adrenal) 

to achieve these effects. Additional in vitro and in vivo studies to address these questions are 

needed. 
Page 9 of 14 

The current data also confirm previous findings (Laws et al., 2009) that though the 

primary ATR metabolite DACT (135 mg/kg; molar equivalent to ATR 200 mg/kg) significantly 

elevates ACTH, CORT, and progesterone as compared to control values at 15 min; however, it 

does so at a magnitude approximately 6-fold less than ATR (200 mg/kg) for ACTH and 

progesterone, and nearly 3-fold less for CORT. This difference in the magnitude of hormone 

changes occurs in spite of the fact that over 60-fold more DACT is found in blood plasma 15 

minutes after a single dose of DACT (135 mg/ kg) than after a single dose of ATR (200 mg/kg; 

Laws et al., 2009). Thus, although it has been postulated that DACT is the active metabolite 

responsible for HPG-axis effects following exposure to ATR, it is unlikely to be responsible for 

the HPA-axis activation observed in this study and others (Laws et al., 2009; Fraites et al., 

2009). Given the rapid metabolism of ATR to DACT, it may be that the parent compound ATR 

and/or its mono-dealkylated intermediary metabolites deisopropyl atrazine (DIA) or deethyl 

atrazine (DEA) are primarily responsible for the effects observed on the HPA-axis after an acute 

exposure, while DACT may play a role in producing the effects on the hypothalamic-pituitarygonadal axis (HPG-axis). 

In conclusion, the current data confirm and extend previous findings that demonstrate 

that ATR increases plasma ACTH (LOEL= 12.5mg/kg), serum CORT (LOEL= 5mg/kg), and 

progesterone (LOEL 5mg/kg) concentrations in a dose-dependent fashion 15 min following a 

single dose. These changes in hormones are robust and reproducible. Though the exact mode/ 

mechanism of action for these changes is unknown, the rapid release of ACTH and subsequent 

activation of adrenal steroidogenesis (i.e. CORT and progesterone release) may reflect 

chlorotriazine-induced changes at the level of the brain and/ or pituitary. 
Page 10 of 14 

References 

Baker, D. B. (1998). Herbicides in drinking water: A challenge for risk communication. Triazine 

Herbicides Risk Assessment. L. F. Ballantine, McFarland, J.E., and Hackett, D.S., American 

Chemical Society, Washinton D.C.: 303-321. 

Balu, K., Holden, P.W., Johnson, L.C., and Cheung, M.W. (1998). Summary of Ciba Crop 

Protection groundwater monitoring study for atrazine and its degradation products in the United 

States. Triazine Herbicides Risk Assessment. L. G. Ballantine, McFarland, J.E., and Hackett, 

D.S., American Chemical Society, Washington D.C.: 227-238. 

Cooper, R. L., S. C. Laws, et al. (2007). "Atrazine and reproductive function: mode and 

mechanism of action studies." Birth Defects Res B Dev Reprod Toxicol 80(2): 98-112. 

Cooper, R. L., T. E. Stoker, et al. (1996). "Effect of atrazine on ovarian function in the rat." 

Reprod Toxicol 10(4): 257-264. 

Cooper, R. L., T. E. Stoker, et al. (2000). "Atrazine disrupts the hypothalamic control of 

pituitary-ovarian function." Toxicol Sci 53(2): 297-307. 

Fan, W., T. Yanase et al. (2007), “Herbicide atrazine activates SF-1 by direct affinity and 

concomitant co-activators recruitments to induce aromatase expression via promoter II.” 

Biochem Biophys Res Commun 355(4): 1012-1018. 

Foradori, C. D., L. R. Hinds, et al. (2009). "Effects of atrazine and its withdrawal on 

gonadotropin-releasing hormone neuroendocrine function in the adult female Wistar rat." Biol 

Reprod 81(6): 1099-1105. 

Foradori, C. D., L. R. Hinds, et al. (2009). "Atrazine inhibits pulsatile luteinizing hormone 

release without altering pituitary sensitivity to a gonadotropin-releasing hormone receptor 

agonist in female Wistar rats." Biol Reprod 81(1): 40-45. 
Page 11 of 14 

Fraites, M. J., R. L. Cooper, et al. (2009). "Characterization of the hypothalamic-pituitaryadrenal axis response to atrazine and metabolites in the female rat." Toxicol Sci 112(1): 88-99. 

Laws, S. C., J. M. Ferrell, et al. (2003). "Pubertal development in female Wistar rats following 

exposure to propazine and atrazine biotransformation by-products, diamino-S-chlorotriazine and 

hydroxyatrazine." Toxicol Sci 76(1): 190-200. 

Laws, S. C., J. M. Ferrell, et al. (2000). "The effects of atrazine on female wistar rats: an 

evaluation of the protocol for assessing pubertal development and thyroid function." Toxicol Sci 

58(2): 366-376. 

Laws, S. C., M. Hotchkiss, et al. (2009). "Chlorotriazine herbicides and metabolites activate an 

ACTH-dependent release of corticosterone in male Wistar rats." Toxicol Sci 112(1): 78-87. 

McMullin, T. S., M. E. Andersen, et al. (2004). "Evidence that atrazine and 

diaminochlorotriazine inhibit the estrogen/progesterone induced surge of luteinizing hormone in 

female Sprague-Dawley rats without changing estrogen receptor action." Toxicol Sci 79(2): 278-

286. 

Pruett, S. B., R. Fan, et al. (2003). "Modeling and predicting immunological effects of chemical 

stressors: characterization of a quantitative biomarker for immunological changes caused by 

atrazine and ethanol." Toxicol Sci 75(2): 343-354. 

Sanderson, J. T., W. Seinen, et al. (2000). "2-Chloro-s-triazine herbicides induce aromatase 

(CYP19) activity in H295R human adrenocortical carcinoma cells: a novel mechanism for 

estrogenicity?" Toxicol Sci 54(1): 121-127. 

Seiler, A., Brenneisen, P., and Breen, D.H. (1992). "Benefits and risks of plant protection 

products- possibilities of protecting drinking water: Case atrazine." Water Supply 10: 31-42. 
Page 12 of 14 

Stoker, T. E., D. L. Guidici, et al. (2002). "The effects of atrazine metabolites on puberty and 

thyroid function in the male Wistar rat." Toxicol Sci 67(2): 198-206. 

Stoker, T. E., S. C. Laws, et al. (2000). "The effect of atrazine on puberty in male wistar rats: an 

evaluation in the protocol for the assessment of pubertal development and thyroid function." 

Toxicol Sci 58(1): 50-59. 

Suzawa, M. and H. A. Ingraham (2008). "The herbicide atrazine activates endocrine gene 

networks via non-steroidal NR5A nuclear receptors in fish and mammalian cells." PLoS One 

3(5): e2117. 
Page 13 of 14 

Figure 1 Dose and time response for ACTH, corticosterone and progesterone concentrations 

following a single dose of atrazine. Male Wistar rats were pre-dosed with vehicle control for 1 

week prior to receiving a single dose of vehicle (methyl cellulose) or), atrazine (5, 50, 100 or 200 

mg/kg, oral gavage). Groups of animals were killed at 5, 15, 30, 60 and or 180 min. post-dosing, 

and ACTH, cortcosterone, and progesterone were measured by RIA. Mean ± SEM (n=10). 

*Significantly different from control at each time point by Kruskal-Wallis Test (non-parametric 

ANOVA) and Dunn’s Multiple Comparison Test (p < 0.05). Mean ACTH ± SEM for controls 

over all time points: 49.51 ±4.25 pg/ml; corticosteorne: 31.08 ±6.04 ng/ml; P4: 0.55 ± 0.07 

ng/ml. 

From: Laws et al., 2009, Toxicol Sci., 112(1):78-87
Page 14 of 14 

Table 1: Hormone Concentrations 15min after a Single Administration of Atrazine 

or DACT 

a

 Mean + SE (n=10 for single block, n=20 for combined blocks unless otherwise noted) 

b

 Dose is equimolar to ATR 200mg/kg 

c

 Indicates different from vehicle control using the LS Means post-hoc test (ANOVA, GLM 

 Procedure, p< 0.05). 

d

 n=9 

e

 n=8 

f n=19 

g

 n=18 

 Dose 

(mg/kg) 

ACTH 

(pg/ml) 

Corticosterone 

(ng/ml) 

Progesterone 

(ng/ml)

Block 1 

ATR 0 64.67 + 3.64 a 46.34 + 5.81 0.58 + 0.06 

ATR 5 75.54 + 5.91 117.64 + 19.56 c 1.09 + 0.16 c 

ATR 12.5 96.29 + 7.55 216.90 + 31.55 c 1.90 + 0.24 c 

ATR 25 127.69 + 11.97 c 281.69 + 27.66 c 2.95 + 0.39 c

ATR 50 135.40 + 18.28 c 284.87 + 33.11 c 3.53 + 0.65 c

ATR 100 332.14 + 76.96 c 364.32 + 29.78 c 5.60 + 0.73 c

ATR 200 479.14 + 99.07 c 356.08 + 23.07 c 6.42 + 0.98 c

DACT 135 b 89.41 + 6.55 c 129.28 + 21.46 c 1.07 + 0.12 c

Block 2 

ATR 0 59.84 + 7.73 d 36.51 + 7.76 0.65 + 0.10 

ATR 5 58.25 + 5.06 d 83.29 + 21.81 c 1.00 + 0.12 

ATR 12.5 99.93 + 9.43 c d 230.24 + 40.12 c 2.29 + 0.39 c

ATR 25 128.66 + 32.67 c d 249.05 + 44.21 c 3.19 + 0.84 c

ATR 50 163.87 + 26.04 c d 339.05 + 35.31 c 4.61 + 0.91 c

ATR 100 318.82 + 89.87 c d 413.09 + 32.58 c 7.28 + 1.06 c

 

ATR 200 439.87 + 93.50 c d 396.60 + 19.47 c 7.60 + 0.55 c

DACT 135 b 64.98 + 5.77 e 113.04 + 27.36 c 1.26 + 0.11c

Blocks 

1 & 2 

Combined

 

ATR 0 62.38 + 4.05 f 41.43 + 4.85 0.62 + 0.06 

ATR 5 67.35 + 4.32 f 100.46 + 14.79 c 1.05 + 0.10 c

ATR 12.5 98.01 + 5.82 c f 223.57 + 24.89 c 2.09 + 0.23 c

ATR 25 128.15 + 16.20 c f 265.37 + 25.66 c 3.07 + 0.45 c

ATR 50 148.89 + 15.55 c f 311.96 + 24.36 c 4.07 + 0.56 c

ATR 100 325.83 + 57.11 c f 388.71 + 22.20 c 6.44 + 0.66 c

ATR 200 460.54 + 66.67 c f 376.34 + 15.41 c 7.01 + 0.57 c

 DACT 135 b 78.55 + 5.23 c g 121.16 + 17.02 c 1.17 + 0.08 c