Abstract
A skeletal examination of fetuses is required in regulatory embryo–fetal development studies. This chapter describes a method of skeletal examination using alizarin staining. All fetuses are removed from the mother by caesarean section before birth. The fetuses are first examined externally. For larger species (rabbit and minipig), an internal examination can be performed on the fresh soft tissues by microdissection. For smaller species, such as the rat and mouse, half of each litter is fixed for internal soft tissue examination. The other half is used for skeletal examination. A rapid examination of the fresh soft tissues is performed before evisceration and fixation. The staining process takes several days. Following staining, all bones are examined from the head to the tail, in ventral and dorsal positions.
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Makris SL et al (2009) Terminology of developmental abnormalities in common laboratory mammals (version 2). Birth Defects Res B Dev Reprod Toxicol 86:227–327
ICH (2009) ICH guideline M3(R2): guidance on nonclinical safety studies for the conduct of human clinical trials and marketing authorization for pharmaceuticals, 11 June 2009
ICH (2005) Harmonised Tripartite Guideline S5(R2). Detection of toxicity to reproduction for medicinal products & toxicity to male fertility
Xu J et al (2005) The antibacterial mechanism of carvacrol and thymol against Escherichia coli. Lett Appl Microbiol 47:174–179
Rivas L et al (2010) Inhibition of verocytotoxigenic Escherichia coli in model broth and rumen systems by carvacrol and thymol. Int J Food Microbiol 139:70–78
Ettayebi K et al (2006) Synergistic effects of nisin and thymol on antimicrobial activities in Listeria monocytogenes and Bacillus subtilis. FEMS Microbiol Lett 18:191–195
OECD (2001) Guideline for the testing of chemicals 414. Prenatal developmental toxicity study
EPA (1998) Health effects test guidelines OPPTS 870.3700. Prenatal developmental toxicity study
FDA (2000) Redbook 2000. Toxicological principles for the safety of food ingredients IV.C.9.b. Guidelines for developmental toxicity studies
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Thanks to the Fetal Pathology team of Ricerca Biosciences.
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Reynaud, L., Jocteur-Monrozier, A. (2013). Skeletal Examination by Alizarin Staining. In: Barrow, P. (eds) Teratogenicity Testing. Methods in Molecular Biology, vol 947. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-131-8_16
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DOI: https://doi.org/10.1007/978-1-62703-131-8_16
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-130-1
Online ISBN: 978-1-62703-131-8
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