Fetal Rabbit Research Articles

Progressive Vascular Functional and Structural Damage in a Bronchopulmonary Dysplasia Model in Preterm Rabbits Exposed to Hyperoxia.

Bronchopulmonary dysplasia (BPD) is caused by preterm neonatal lung injury and results in oxygen dependency and pulmonary hypertension. Current clinical management fails to reduce the incidence of BPD, which calls for novel therapies. Fetal rabbits have a lung development that mimics humans and can be used as a translational model to test novel treatment options. In preterm rabbits, exposure to hyperoxia leads to parenchymal changes, yet vascular damage has not been studied in this model. In this study we document the early functional and structural changes of the lung vasculature in preterm rabbits that are induced by hyperoxia after birth. Pulmonary artery Doppler measurements, micro-CT barium angiograms and media thickness of peripheral pulmonary arteries were affected after seven days of hyperoxia when compared to controls. The parenchyma was also affected both at the functional and structural level. Lung function testing showed higher tissue resistance and elastance, with a decreased lung compliance and lung capacity. Histologically hyperoxia leads to fewer and larger alveoli with thicker walls, less developed distal airways and more inflammation than normoxia. In conclusion, we show that the rabbit model develops pulmonary hypertension and developmental lung arrest after preterm lung injury, which parallel the early changes in human BPD. Thus it enables the testing of pharmaceutical agents that target the cardiovascular compartment of the lung for further translation towards the clinic.

Differential gene expression profiles in foetal skin of Rex rabbits with different wool density

<p>This study investigated the mechanisms controlling hair follicle development in the Rex rabbit. The Agilent rabbit gene expression microarray was used to determine differentially expressed genes in Rex rabbit foetuses with different wool densities. The expression patterns of selected differentially-expressed genes were further investigated by quantitative real-time PCR. Compared to low wool density rabbits, 1342 differentially expressed probes were identified in high wool density rabbits, including 950 upregulated probes and 392 downregulated probes. Gene ontology analysis revealed that the most upregulated differentially expressed probes belonged to receptors and the most downregulated differentially expressed probes belonged to DNA binding molecules. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the differentially expressed probes were mainly involved in the sonic hedgehog (Shh) and Eph signalling pathways. The results also suggest that transforming growth factor-beta 1, growth hormone receptor, and the keratin-associated protein 6.1 genes, as well as the Shh and Eph signalling pathways, may be involved in the regulation of hair follicle developmental in Rex rabbits.</p>

Evaluation of the protective effect of thiamine pyrophosphate based on the biochemical analysis of rabbit foetuses at 30 days of gestation

This study evaluated the effects of thiamine pyrophosphate (PPT) on the biochemical profiles of full-term rabbit foetuses that were subjected to experimental ischemia followed by 24h reperfusion. A total of 16 gestating rabbit dams were divided into two groups, one of which was treated by administering PPT and subjected to a process ischemia. During this interval, fetal blood samples were drawn from each dam (in the ischemia group) at 0, 15 and 45min. Ischemia for 15 and 45min was not associated with changes in lactate levels of the Ischemia group foetuses. However, in the foetuses in the reperfusion groups without PPT lactate levels were significantly higher after 15 and 45min of arterial occlusion compared to time zero. These results demonstrate that PTT alters some acute and some longer-term biochemical outcomes of uterine ischemia perhaps important in preserving energy metabolism under hypoxic conditions.

Why vessels do matter in pulmonary disease

In addition to structural abnormalities of the diaphragm, congenital diaphragmatic hernia (CDH) presents with severe hypoplasia of lung tissue involving both the pulmonary vasculature and the airways. These developmental defects...

Maternal inflammation leads to impaired glutamate homeostasis and up-regulation of glutamate carboxypeptidase II in activated microglia in the fetal/newborn rabbit brain

Astrocyte dysfunction and excessive activation of glutamatergic systems have been implicated in a number of neurologic disorders, including periventricular leukomalacia (PVL) and cerebral palsy (CP). However, the role of chorioamnionitis on glutamate homeostasis in the fetal and neonatal brains is not clearly understood. We have previously shown that intrauterine endotoxin administration results in intense microglial ‘activation’ and increased pro-inflammatory cytokines in the periventricular region (PVR) of the neonatal rabbit brain. In this study, we assessed the effect of maternal inflammation on key components of the glutamate pathway and its relationship to astrocyte and microglial activation in the fetal and neonatal New Zealand white rabbit brain. We found that intrauterine endotoxin exposure at gestational day 28 (G28) induced acute and prolonged glutamate elevation in the PVR of fetal (G29, 1day post-injury) and postnatal day 1 (PND1, 3days post-injury) brains along with prominent morphological changes in the astrocytes (soma hypertrophy and retracted processes) in the white matter tracts. There was a significant increase in glutaminase and N-Methyl-d-Aspartate receptor (NMDAR) NR2 subunit expression along with decreased glial L-glutamate transporter 1 (GLT-1) in the PVR at G29, that would promote acute dysregulation of glutamate homeostasis. This was accompanied with significantly decreased TGF-β1 at PND1 in CP kits indicating ongoing neuroinflammation. We also show for the first time that glutamate carboxypeptidase II (GCPII) was significantly increased in the activated microglia at the periventricular white matter area in both G29 and PND1 CP kits. This was confirmed by in vitro studies demonstrating that LPS activated primary microglia markedly upregulate GCPII enzymatic activity. These results suggest that maternal intrauterine endotoxin exposure results in early onset and long-lasting dysregulation of glutamate homeostasis, which may be mediated by impaired astrocyte function and GCPII upregulation in activated microglia.

MORPHOMETRIC BRAIN MEASUREMENT OF PRENATAL ULTRASOUND-INDUCED RABBIT FETUS USING MICRO-COMPUTED TOMOGRAPHY (MICRO-CT)

Prenatal ultrasound is widely used and became an integral part of the clinical practice, particularly in obstetrics and gynecology. However, the advances in the capability of ultrasound equipment nowadays, trigger a greater image processing power, hence might as well increase the fetal exposure. Hence premises the morphometric brain measurement to be assessed in this study as an evidence of the ultrasound interruptions during the fetal neuronal development. This research randomly analyzed a total of 18 ultrasound-induced rabbit fetuses for fetal brain volume and surface after being exposed prenatally to the ultrasound exposure (duration=60 minutes; frequency = 7.09 MHz; spatial peak temporal average intensity (ISPTA) = 49.4 W/cm2; power = 56.0 W; thermal index (TI) = 0.2; mechanical index (MI) = 1.0). The fetuses were analyzed for morphometric brain measurement of brain volume and surface using Skyscan™ 1176 Micro-computed Tomography (Micro-CT). There were significant differences in the measurement of brain volume and surface at the 2nd and 3rd stage of gestation (P < 0.05). Results suggested that there are significant differences in the brain volume and surface between the controls and the 2nd and 3rd stage of gestation. There are also a significant reduction in the brain volume and surface in the exposed groups at all stages of gestation (P < 0.05).

Nitric oxide synthase and changes in oxidative stress levels in embryonic kidney observed in a rabbit model of intrauterine growth restriction

This work aimed to study the effect of uteroplacental circulation restriction on endothelial kidney damage in a fetal rabbit model. New Zealand rabbits were subjected to 40% to 50% of uteroplacental artery ligation at day 25 of pregnancy. After 5 days, surviving fetuses were harvested by cesarean section. The gene and protein expressions of selected enzymes associated with nitric oxide production and oxidative stress were analyzed in fetal kidney homogenates. The placenta weight (6.06 ± 0.27, p < 0.0319) and fetal body (19.90 ± 1.03, p < 0.0001) were significantly reduced in the uteroplacental circulation restriction group. The kidneys from restricted fetuses presented a mild vascular congestion and glomerular capillary congestion, without inflammation or hypertrophy. We found endothelial nitric oxide synthase phosphorylation inhibition (0.23 ± 0.13, p < 0.012) and arginase-2 (0.29 ± 0.14, p < 0.023) protein induction in fetal kidneys of the circulation restriction group. Finally, the kidneys from circulation-restricted fetuses showed increased inducible nitric oxide synthase messenger RNA (mRNA) (2.68 ± 0.24, p < 0.01) and reduced heme oxygenase-1 mRNA (23 ± 1.3, p < 0.003), with increased reactive oxygen species (1.69 ± 0.09, p < 0.001) and nitrotyrosine protein (1.74 ± 0.28, p < 0.003) levels, without changes in Nox mRNA. We describe significant deregulation of vascular activity and oxidative damage in kidneys of fetal rabbits that have been exposed to restriction of the uterine circulation. © 2016 John Wiley & Sons, Ltd.

Transplacental sildenafil rescues lung abnormalities in the rabbit model of diaphragmatic hernia

IntroductionThe management of congenital diaphragmatic hernia (DH) would benefit from an antenatal medical therapy, which addresses both lung hypoplasia and persistent pulmonary hypertension. We aimed at evaluating the pulmonary effects...

Antenatal BAY 41-2272 reduces pulmonary hypertension in the rabbit model of congenital diaphragmatic hernia

Infants with congenital diaphragmatic hernia (CDH) fail to adapt at birth because of persistent pulmonary hypertension (PH), a condition characterized by excessive muscularization and abnormal vasoreactivity of pulmonary vessels. Activation of soluble guanylate cyclase by BAY 41-2272 prevents pulmonary vascular remodeling in neonatal rats with hypoxia-induced PH. By analogy, we hypothesized that prenatal administration of BAY 41-2272 would improve features of PH in the rabbit CDH model. Rabbit fetuses with surgically induced CDH at day 23 of gestation were randomized at day 28 for an intratracheal injection of BAY 41-2272 or vehicle. After term delivery (day 31), lung mechanics, right ventricular pressure, and serum NH2-terminal-pro-brain natriuretic peptide (NT-proBNP) levels were measured. After euthanasia, lungs were processed for biological or histological analyses. Compared with untouched fetuses, the surgical creation of CDH reduced the lung-to-body weight ratio, increased mean terminal bronchial density, and impaired lung mechanics. Typical characteristics of PH were found in the hypoplastic lungs, including increased right ventricular pressure, higher serum NT-proBNP levels, thickened adventitial and medial layers of pulmonary arteries, reduced capillary density, and lower levels of endothelial nitric oxide synthase. A single antenatal instillation of BAY 41-2272 reduced mean right ventricular pressure and medial thickness of small resistive arteries in CDH fetuses. Capillary density, endothelial cell proliferation, and transcripts of endothelial nitric oxide synthase increased, whereas airway morphometry, lung growth, and mechanics remained unchanged. These results suggest that pharmacological activation of soluble guanylate cyclase may provide a new approach to the prenatal treatment of PH associated with CDH.

Pulmonary transcriptome analysis in the surgically induced rabbit model of diaphragmatic hernia treated with fetal tracheal occlusion.

ABSTRACTCongenital diaphragmatic hernia (CDH) is a malformation leading to pulmonary hypoplasia, which can be treated in utero by fetal tracheal occlusion (TO). However, the changes of gene expression induced by TO remain largely unknown but could be used to further improve the clinically used prenatal treatment of this devastating malformation. Therefore, we aimed to investigate the pulmonary transcriptome changes caused by surgical induction of diaphragmatic hernia (DH) and additional TO in the fetal rabbit model. Induction of DH was associated with 378 upregulated genes compared to controls when allowing a false-discovery rate (FDR) of 0.1 and a fold change (FC) of 2. Those genes were again downregulated by consecutive TO. But DH+TO was associated with an upregulation of 157 genes compared to DH and controls. When being compared to control lungs, 106 genes were downregulated in the DH group and were not changed by TO. Therefore, the overall pattern of gene expression in DH+TO is more similar to the control group than to the DH group. In this study, we further provide a database of gene expression changes induced by surgical creation of DH and consecutive TO in the rabbit model. Future treatment strategies could be developed using this dataset. We also discuss the most relevant genes that are involved in CDH.

Excessive Reversal of Epidermal Growth Factor Receptor and Ephrin Signaling Following Tracheal Occlusion in Rabbit Model of Congenital Diaphragmatic Hernia.

Congenital diaphragmatic hernia (CDH) causes severe pulmonary hypoplasia from herniation of abdominal contents into the thorax. Tracheal occlusion (TO) for human CDH improves survival, but morbidity and mortality remain high, and we do not fully understand the cellular pathways and processes most severely impacted by CDH and TO. We created a left diaphragmatic hernia (DH) in rabbit fetuses with subsequent TO and collected left lung sections for NextGen mRNA sequencing. DH, TO, and DHTO fetuses had comparable body and organ growth to control except for lower lung weights in DH (p<0.05). Of 13,687 expressed genes, DHTO had 687 differentially expressed genes compared to DH, but no other group-group comparison had more than 10. Considering genes in combination, many of the genes reduced in DH were more highly expressed in DHTO than in control. Benchmarking fetal rabbit lung gene expression to published lung development data, both DH and DHTO lungs were more highly correlated with the gene expression of immature lung. DNA synthesis was upregulated in DHTO compared to DH and ribosome and protein synthesis pathways were downregulated. DH reduced total and epithelial cell proliferation by half and two-thirds respectively, and DHTO increased proliferation by 2.5 and 3.4-fold respectively. Signaling pathways downregulated by DH and upregulated in DHTO were epidermal growth factor receptor signaling, ephrin signaling, and cell migration; however, levels of ephrin and EGFR signaling in DHTO exceeded that of control. Identification and inhibition of the ligands responsible for this dysregulated signaling could improve lung development in CDH.

The Effect of Transplacental Administration of Glucagon-Like Peptide-1 on Fetal Lung Development in the Rabbit Model of Congenital Diaphragmatic Hernia

Objective: Glucagon-like peptide-1 (GLP-1) increases surfactant protein expression in type 2 pneumocytes. Herein, we determine if transplacental GLP-1 treatment accelerates lung growth in the fetal rabbit model of congenital diaphragmatic hernia (DH). Methods: Time-mated does had an induction of DH on day 23 followed by daily GLP-1 or placebo injection until term. At that time, the does were weighed, fetal blood was obtained for GLP-1 assay, and the lungs were dissected. Fetal outcome measures were lung-to-body-weight ratio (LBWR), morphometry, and Ki67 and surfactant protein B (SPB) expression. Results: Maternal weight loss in the GLP-1 group was 7.1%. Fetal survival was lower in GLP-1 fetuses compared to placebo controls (27/85, 32% vs. 35/57, 61%; p < 0.05). Fetal GLP-1 levels were increased 3.6-fold. The LBWR of GLP-1 DH fetuses fell within the range of DH placebo fetuses (1.166 ± 0.207% vs. 1.312 ± 0.418%), being significantly lower than that of placebo-exposed unoperated fetuses (2.280 ± 0.522%; p < 0.001). GLP-1 did not improve airway morphometry. GLP-1 DH lungs had a reduced adventitial and medial thickness within the range of controls, and lesser muscularization of vessels measuring 30-60 µm. There were no differences in Ki67 and SPB expression. Conclusion: GLP-1 at this dosage improves peripheric pulmonary vessel morphology in intra-acinar vessels with no effect on airway morphometry but with significant maternal and fetal side effects. Thus, it is an unlikely medical strategy.

Leptin Gene Expression in Rabbits During Pregnancy and Fetal Life

Leptin may act as the critical link between adipose tissue and reproduction. Although considerable progress has been achieved in understanding the reproductive actions of leptin, much work is needed for understanding its physiological role. Till now, no data has been published about the distribution and expression levels of leptin in the rabbit maternal adipose tissue, placenta, and various fetal rabbit tissues during pregnancy and postpartum. Our results indicated that circulating leptin levels in rabbit serum during pregnancy were significantly higher than in postpartum and non pregnant rabbits. Furthermore, leptin showed positive correlations with body weight and estrogen and negative correlation with progesterone in pregnant and postpartum rabbits. RT-PCR verified the presence of RNAs encoding leptin in the rabbit maternal perirenal adipose tissue, placenta, and several fetal tissues; including brain, liver, adipose tissue, and bone. The relative abundance of leptin RNA in rabbit maternal adipose was significantly higher at 20 th day of pregnancy than that of non pregnant rabbits, while it was significantly decreased at 2 nd day after parturition. No significant changes in the placental leptin RNA levels were noticed in pregnant rabbits at 10 th , 20 th , and 30 th day of pregnancy. The relative abundance of fetal leptin transcripts at day 30 th of pregnancy was in the order of liver> bone ? adipose tissue > brain. The present study provides new evidence for the distribution and expression levels of leptin in the rabbit maternal and fetal tissues during pregnancy and supports the importance of leptin in reproductive physiology and fetal development.

Inhibition ofPasteurella multocidaAdhesion to Rabbit Respiratory Epithelium Using Lectins

This study aimed to evaluate the ability of a panel of lectins to inhibit the ability of Pasteurella multocida to adhere to and affect the rabbit respiratory epithelium. Nasal septa from rabbit fetuses were cultured with various lectins before the addition of P. multocida. The percentage of bacteria adhering to the epithelium was evaluated semiquantitatively by indirect immunoperoxidase (IIP) staining. The goblet cells (GCs) were counted in semithin sections stained with toluidine blue and served as the main morphological criterion to evaluate the inhibitory effect of the lectins. The lectins PNA, WGA, RCA120, and DBA significantly inhibited the adhesion of P. multocida to the ciliated epithelium (P < 0.05) and prevented the pathogen-induced increase in the number of GCs (P < 0.05) compared with those of positive control tissues. In addition, VVA, SJA, UEA I, DSL, SBA, and ECL significantly inhibited the increase in GCs compared with that of the control tissues. The results suggest that less aggressive therapeutic strategies, such as treatment with lectins, may represent alternative approaches to control bacterial respiratory infections.

317: Intratracheal delivery of amniotic fluid derived stem cells (AFSc) in perfluorcarbon as compared to tracheal occlusion (TO)

Perfluoro-octyl-bromide (PFOB) and amniotic fluid derived stem cells (AFSc) can be used to modulate lung development. We assessed the compatibility of AFSc with PFOB and their effects on fetal rabbit lungs in vivo as compared to tracheal occlusion (TO).

Intrapulmonary instillation of perflurooctylbromide improves lung growth, alveolarization, and lung mechanics in a fetal rabbit model of diaphragmatic hernia.

Fetal tracheal occlusion of hypoplastic rabbit lungs results in lung growth and alveolarization although the surfactant protein messenger RNA expression is decreased and the transforming growth factor-β pathway induced. The prenatal filling of healthy rabbit lungs with perfluorooctylbromide augments lung growth without suppression of surfactant protein synthesis. We hypothesizes that Intratracheal perfluorooctylbromide instillation improves lung growth, mechanics, and extracellular matrix synthesis in a fetal rabbit model of lung hypoplasia induced by diaphragmatic hernia. On day 23 of gestation, DH was induced by fetal surgery in healthy rabbit fetuses. Five days later, 0.8ml of perfluorooctylbromide (diaphragmatic hernia-perfluorooctylbromide) or saline (diaphragmatic hernia-saline) was randomly administered into the lungs of previously operated fetuses. After term delivery (day 31), lung mechanics, lung to body weight ratio, messenger RNA levels of target genes, assessment of lung histology, and morphological distribution of elastin and collagen were determined. Nonoperated fetuses served as controls. Fetal instillation of perfluorooctylbromide in hypoplastic lungs resulted in an improvement of lung-to-body weight ratio (0.016 vs 0.013 g/g; p = 0.05), total lung capacity (23.4 vs 15.4 μL/g; p = 0.03), and compliance (2.4 vs 1.2 mL/cm H2O; p = 0.007) as compared to diaphragmatic hernia-saline. In accordance with the results from lung function analysis, elastin staining of pulmonary tissue revealed a physiological distribution of elastic fiber to the tips of the secondary crests in the diaphragmatic hernia-perfluorooctylbromide group. Likewise, messenger RNA expression was induced in genes associated with extracellular matrix remodeling (matrix metalloproteinase-2, tissue inhibitor of metalloproteinase-1, and tissue inhibitor of metalloproteinase-2). Surfactant protein expression was similar in the diaphragmatic hernia-perfluorooctylbromide and diaphragmatic hernia-saline groups. Distal airway size, mean linear intercept, as well as airspace and tissue fractions were similar in diaphragmatic hernia-perfluorooctylbromide, diaphragmatic hernia-saline, and control groups. Fetal perfluorooctylbromide treatment improves lung growth, lung mechanics, and extracellular matrix remodeling in hypoplastic lungs, most probably due to transient pulmonary stretch, preserved fetal breathing movements, and its physical characteristics. Perfluorooctylbromide instillation is a promising approach for prenatal therapy of lung hypoplasia.

PO-0501 Glibenclamide Closes Neonatal Ductus Arteriosus In Rats

Background Treatment for patent ductus arteriosus (PDA) in premature infants may fail following perinatal indomethacin exposure. Glibenclamide, a sulfonylurea, constricts the fetal rabbit ductus in vitro , and fetal rat ductus in vivo . Clinical doses for diabetes include 0.1 mg/kg (adults) and 1 mg/kg (children with neonatal diabetes). Aims To show ductus constriction acceleration with Glibenclamide in newborn rats as a model of treatment for premature PDA. Methods Glibenclamide (1 mg/Kg) was injected intraperitoneally (IP) to newborn Wistar rats immediately after cesarian section, and the ductus diameter was studied at 60 min with rapid whole-body freezing, by cutting on a freezing microtome, and measurements performed on a microscope using a micrometre. Two near-term rat models were studied on the 21st day. In a chronic fetal indomethacin-exposure model, mother rats were treated with indomethacin (10 mg/kg, gavage) for two days before birth. In a hypoxia model, neonates were incubated in 8% oxygen. In a premature model, rats were delivered on the 19th day (two days before term) and incubated in 80% oxygen. Results In these three models, neonatal ductus constricted slowly. Glibenclamide 1 mg/kg, caused accelerated constriction and the effects were dose-dependent. Glibenclamide (1mg/kg, IP) was associated with hypoglycemia, which was controlled with 50% glucose via gavage. Conclusions Glibenclamide (1 mg/kg, IP) constricts the neonatal ductus in 60 min in three rat models. Hypoglycemia was controlled with glucose, indicating its usefulness in the treatment of PDA in premature infants.

Leptin Gene Expression in Rabbits during Pregnancy And Fetal Life

Leptin may act as the critical link between adipose tissue and reproduction.Although considerable progress has been achieved in understanding the reproductive actions of leptin, much work is needed for understanding its physiological role. Till now, no data has been published about the distribution and expression levels of leptin in the rabbit maternal adipose tissue, placenta, and various fetal rabbit tissues during pregnancy and postpartum. Our results indicated that circulating leptin levels in rabbit serum during pregnancy were significantly higher than that in postpartum and non pregnant rabbits. Furthermore, leptin showed positive correlations with body weight in pregnant and postpartum rabbits. RT-PCR verified the presence of RNAs encoding leptin in the rabbit maternal perirenal adipose tissue, placenta, and several fetal tissues; including brain, liver, adipose tissue, and bone. The relative abundance of leptin RNA in rabbit maternal adipose tissue was significantly higher at 20th day of pregnancy than that of non pregnant rabbits, while it was significantly decreased at 2nd day after parturition. No significant changes in the placental leptin RNA levels were noticed in pregnant rabbits at 10th, 20th, and 30th day of pregnancy. The relative abundance of fetal leptin transcripts at day 30th of pregnancy was in the order of liver> bone ≥ adipose tissue > brain. The present study provides new evidence for the distribution and expression levels of leptin in the rabbit maternal and fetal tissues during pregnancy and supports the importance of leptin in reproductive physiology and fetal development.

OP18.09: In vivo assessment of tissue‐specific metabolic changes in the fetal rabbit brain associated with intrauterine growth restriction (IUGR)

<scp>OP</scp>18.09: In vivo assessment of tissue‐specific metabolic changes in the fetal rabbit brain associated with intrauterine growth restriction (<scp>IUGR</scp>)

English

The present study was carried out to determine the effects of aflatoxinB1 (AFB1) on the skeletal system of the fetuses of balady rabbits. The female animals were divided into three groups, one control and two treated, the control group contained four dams, the intoxicated group with 0.1 mg/kg contained six dams while the intoxicated with 0.05 mg/kg contained three dams. A dose of 0.1 and 0.05 mg/kg/day AFB1 was administered by gastric intubation to pregnant rabbits on the 6th-21st day of pregnancy. The fetuses were obtained through the uterine incisionat different stages of gestation according to the group. The lengths and weights of the fetuses as well as absolute organs weights were measured, revealing the statistically significant differences between the control and intoxicated with 0.05 mg/kg group at 29th day of gestation (p&lt;0.001) while there were non-significant decrease of control and intoxicated with 0.1 mg/kg at 22nd day of gestation. The observed gross anomalies included wrinkled skin, enlarged eye socket and microphthalmic eyes in both groups. The heart of treated group showed reduction in size with wide ventricular lumen and shallow inter ventricular groove in intoxicated group with a dose of 0.05 mg/kg AFB1. Regarding the skeletal anomalies, there were incomplete ossification in some of the skull bones, the laminae of the vertebral arches throughout the vertebral column remain cartilaginous. The sternum was incompletely ossified. Most of the appendicular skeleton bones were grossly shorter and remained in cartilaginous state. &nbsp; Key words: Aflatoxin B1 (AFB1), rabbits, teratogenicity, ossification.