Abstracts / Toxicology Letters 189S (2009) S57S273 S143

Reproduction Toxicology
J01
Ultrastructural studyof aminoglycosides (gentamicin) anduo-
roquinolone (ooxacin) antibiotics effect ontestis tissue inrats:
Light and transmission electron microscopic study
Arash Khaki
1,
, Farnaz Rad saeid
1
, Mohammad Hamadeh
2
1
Islamic Azad University Tabriz Branch, Veterinary Pathology, Tabriz,
Iran, Islamic Republic of Iran,
2
University of Saarland Germany,
Obstetrics and Gynecology, Hamburg, Germany
Aim: Theaimof this studywas toinvestigatethecomparativeeffects
of aminoglycosides and uoroquinolones on testis tissue in rats.
Material and methods: Thirty male Wister rats were randomly
divided into control (n=10) and experimental (n=20) groups. The
experimental groups were subdivided into two groups of ten. Each
received 5mg/kg (IP) Gentamicin and 72mg/kg (IP) Ooxacin daily
for 14 days, respectively; however, the control group just received
vehicle(IP). Onthefourteenthday, rats werekilledandtestis tissues
were also prepared for light and electron microscopic study.
Results: All animals exposed to drugs were seen a depletion of
germcells, germinal cells necrosis, especiallyinspermatogonia, and
Leydig cells had an abnormal broblast-like appearance, Abnormal
spacebetweenneighbor sertoli cells, mitochondriawerelost cristae
andvacuolated(noneenergized), lyzosomeseenmoreincytoplasm
of sertoli cells, nucleus of moiled cell was done heterochromatin.
Conclusion: Gentamicin and Ooxacin have negative effects on
testis architecture and germinal cells damages in rats. However,
these side effects are less seen in the Ooxacin group. Therefore, it
is recommended that usage of this drug has fewer side effects on
male fertility.
Keywords: Gentamicin; Ooxacin; Testis; Rat
doi:10.1016/j.toxlet.2009.06.806
J02
Exposure assessments in nonclinical reproductive toxicology
studies using toxicokinetics, including a novel approach for
monitoring pre-/postnatal study exposures
Thomas Grizzle
Toxicology Services Inc., Pharmaceutical Development, Chapel Hill
North Carolina, United States
The reproductive safety prole for many candidate pharmaceu-
ticals is incomplete without an assessment of rat and rabbit
plasma/serumconcentrations of the test substance over time, along
with the generation of toxicokinetic (TK) data. Developmental toxi-
cology studies, where dosing typically occurs fromimplantation of
the embryo through closure of the hard palate, utilizes TK assess-
ments on the rst and last days of dosing in concurrently dosed
satellite dams and does. Dosing can be extended an extra day in
the pregnant satellite animals for a third blood collection, this time
fromfetuses, since a substance that tests negative for teratogenicity
could be due to a lack of fetal exposure to the substance. For pre-
/post-natal studies, additional bioanalytical assays are validated for
a new matrix, milk, and dams are milked at various intervals. A
novel approach has been developed which avoids this additional
validation, and instead uses plasma from culled pups. On postna-
tal day (PND)4 (birth=PND0), litter sizes are normalized within
a dose group to 4M and 4F pups/dam, a process in which culled
pups are identied. A timed sacrice is then conducted on PND4,
n=3 per time point, and a plasma concentration vs. time curve is
thus constructed and TK parameters derived. Exposure data from
plasma/serum of suckling pups is superior to milk concentration
data because the test substance in milk might have poor bioavail-
ability. In summary, TK in reproductive toxicology studies are a
necessary part of safety assessments, and an improved method for
assessing TK in pre-/post-natal studies is described.
doi:10.1016/j.toxlet.2009.06.807
J03
Teratogenic effects of metabolically activated trimethadione in
zebrash embryos (Danio rerio)

Stefan Weigt
1,
, Nicole Hbler
1
, Friedrich von Landenberg
1
,
Thomas Braunbeck
2
, Thomas Broschard
1
1
Merck KGaA, Institute of Toxicology, Darmstadt, Germany,
2
University of Heidelberg, Department of Zoology, Heidelberg,
Germany
The Danio rerio embryo test with metabolic activation (mDarT)
was developed to assess the teratogenic effects of proteratogens.
Induced rat liver microsomes (RLM) are used as a mammalian
metabolic activation system (MAS), since they contain various
cytochrome P450 (CYP) isoforms in high concentrations. The
human proteratogen Trimethadione (TMO) is an anticonvulsant
usedinthetreatment of epilepsy. If administeredduringpregnancy,
the fetal trimethadione syndrome may result causing facial dys-
morphism, cardiac defects, growth and mental retardation. TMO is
metabolically N-demethylated into Dimethadione, mainly by CYP
2E1. The teratogenic potential of TMOwas investigated in zebrash
embryos (20 eggs/group) with RLM under moderate agitation at
32

C for 1h, beginning at the latest 2hpf (hours post fertilization).

The negative control (TMO alone) and the vehicle control (MAS
alone) wereincubatedinparallel. After incubation, theshembryos
were individualized in 24-well plates lled with sh medium and
kept at 26

C for the next 72h with a 12-h-lightdark cycle. Ter-

atogenicity was scored at 24, 48 and 72hpf using standardized
morphological endpoints. All the controls fullled the acceptance
criteria of 10% impaired embryos. While TMO did not show any
teratogenic activity up to 15mM, statistically signicant terato-
genic activity was observed in the presence of RLM. The number
of embryos with teratogenic effects increased with increasing con-
centrations of TMO. It is concluded that the teratogenic potential
of TMO can be detected in zebrash embryos only in the presence
of a metabolic activation system.

Selected for Oral Presentation.

doi:10.1016/j.toxlet.2009.06.808