Monograph on the Potential Human Reproductive and ... - OEHHA

the 1.8 ppm group was not considered to be treatmentrelated
by the study authors because no dose–response
relationship was observed. There was no effect on
anogenital distance in F1 or F2 males or females on
PND 0. Anogenital distance was also unaffected in F2
males and F1 and F2 females on PND 21. Anogenital
distance adjusted for body weight was reduced in F1
males from the 300 and 3500 ppm groups on PND 21.
Based on the lack of effect on anogenital distance at birth
and inconsistencies between generations, the study
authors did not consider the decreases in anogenital
distance in F1 males to be treatment-related. An increase
in anogenital distance in F2 females from the 0.018 ppm
group on PND 0 was not considered to be treatmentrelated
by the study authors. Preputial separation
(absolute age and adjusted for body weight on day of
acquisition) was delayed in parental and retained F 1
males of the 3500 ppm group. When adjusted for PND 30
body weight, preputial separation was delayed in
retained but not parental F1 males from the 3500 ppm
group. Data for preputial separation adjusted for body
weight on day of acquisition are shown in Table 99. Body
weights on day of vaginal opening were lower in F1
females from the 3500 ppm group. Day of vaginal
opening was accelerated in the 3500 ppm group if
adjusted for PND 21 body weight, but not body weight
on the day of acquisition. Due to the lack of effect when
adjusted for body weight on day of acquisition, the study
authors did not consider effects on vaginal opening to be
treatment-related.
Shown in Table 99 are significant organ weight effects
relative to body weight. Dose-related organ weight
changes in F 1 weanlings that were considered to be
treatment-related by study authors included decreased
absolute and relative (to body or brain weight) spleen
and paired testes weights at 3500 ppm. Treatment-related
absolute organ weight changes in F2 weanlings included
decreased weights of spleen, paired testes, and seminal
vesicles with coagulating glands in the 3500 ppm group.
Changes in organ weights relative to body weight in F2
weanlings included decreased spleen weight in males
and females and increased relative left kidney weight in
3500 ppm males. Treatment-related changes in organ
weight relative to brain weight in F2 weanlings were
decreased spleen weight in both sexes and decreased
paired testes weight at 3500 ppm and seminal vesicles
with coagulating glands at 300 and 3500 ppm. Other
organ weight effects (e.g., affecting epididymides,
thymus, brain, ovaries, and/or uterus with cervix and
vagina weights) were not considered to be dose-related
due to lack of dose–response relationships or no
consistent effects across generations. Included in Table 99
are significant organ weight effects relative to body
weight. Significant organ weight effects relative to brain
weight were included in Table 99 when the organ weight
effect was significant only when normalized for brain
weight. The study authors reported no gross findings in
F 1 or F 2 weanlings. The incidence of undescended
bilateral testes was increased in F 1 and F 2 weanling
males of the 3500 ppm group. The incidence of hepatic
cytoplasm alteration (clear hepatocellular cytoplasm,
slightly more basophilic cytoplasm, and/or minute
vacuoles) was apparently increased in F1 males from
the 300 and 3500 ppm groups and F1 females and F2
males from the 3500 ppm group. The incidence of
Birth Defects Research (Part B) 83:157–395, 2008
BISPHENOL A
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seminiferous tubule hypoplasia was increased in F 1 and
F 2 weanlings from the 3500 ppm group. [Another
histopathological finding that appeared to be possibly
increased in weanlings from the 3500 ppm group was
unilateral hydronephrosis in F1 males. It did not appear
that histopathological data were statistically analyzed.]
Effects of 17b-estradiol in males were delayed preputial
separation, reduced anogenital distance at weaning but not
at birth, decreased weights of testes, epididymides, and
seminal vesicles with coagulating gland, and increased
incidence of seminiferous tubule hypoplasia and undescended
testis. Effects of 17b-estradiol in female mice were
accelerated vaginal patency, increased uterus with cervix
and vagina weight, fluid filled/enlarged uterus, enlarged/
thickened vagina, increased vaginal epithelial keratinization,
and prolonged gestation. Reproductive effects in the
17b-estradiol group included decreased fertility, increased
stillbirth, reduced live pups per litter, and increased dead
pups.
The study authors identified bisphenol A NOELs of
30 ppm (B5 mg/kg bw/day) for systemic effects,
300 ppm (B50 mg/kg bw/day) for developmental toxicity,
and 300 ppm (B50 mg/kg bw/day) for reproductive
toxicity.
Strengths/Weaknesses: Strengths include the large
number and range of doses examined, the rigor with
which the study was performed, the large sample size in
each group, the number of additional animals per litter
that were retained and examined, the use of a concurrent
estrogenic positive control group, and the thoroughness
of the histologic evaluation.
Utility (Adequacy) for CERHR Evaluation Process:
This study is adequate and of high utility for the
evaluation process.
4.2.3.3 Fish and invertebrates: Although studies in
fish and invertebrates may be important for understanding
mechanisms of action and environmental
impact, the Panel views these studies as not useful for
the evaluation process.
Kwak et al. (2001), supported by the Korean Ministry
of the Environment, exposed adult male swordtail fish
(Xiphophorus helleri) to bisphenol A 0, 0.4, 2, or 10 ppm
[mg/L] for 72 hr (n 5 20 fish/group). [No information on
purity or culture ware was provided.] [Nonylphenol
was also studied but will not be discussed here.] At the
end of the exposure period, the fish were killed and
livers were removed for measurement of vitellogenin.
Testes of 10 fish/group were processed for flow
cytometry by preparation of single cell suspensions
stained with annexin V-fluorescein isothiocyanate and
propidium iodide to detect necrosis and apoptosis.
TUNEL staining was used to confirm apoptosis in testis
sections. In a second experiment, juvenile male fish (30
days old) were exposed to bisphenol A in water at 0, 0.2,
2 and 20 ppb [lg/L] for 60 days, after which body length
and sword length were measured. [The sword is a
portion of the caudal fin that elongates as a secondary
sex characteristic.] Statistical analysis used ANOVA
followed by least significant difference test. Hepatic
vitellogenin was increased by bisphenol A [data were
not shown]. Apoptosis was increased in testes from fish
exposed to bisphenol A at 10 ppm [mg/L] by TUNEL
assay. [Flow cytometry was said to be more sensitive,
but data did not appear to have been statistically
analyzed.] Sword growth was decreased by bisphenol A