Total Malformations Research Articles

Unveiling the molecular mechanisms and developmental consequences of mercury (Hg) toxicity in zebrafish embryo-larvae: A comprehensive approach

Mercury (Hg) is a global contaminant affecting aquatic ecosystems' health. Chronic exposure to Hg has shown that the normal development of zebrafish embryo-larvae is affected. However, the molecular mechanisms behind the toxicity of Hg on fish embryonic development are still poorly understood. This work aimed to investigate the effects of Hg exposure on zebrafish embryo-larvae using a combined approach at individual (mortality, embryo development and locomotor behavior) and biochemical (neurotoxicity and oxidative stress enzymatic activities and protein phosphatase expression) levels. The Fish Embryo Toxicity assay followed the Organization for Economic Cooperation and Development Guideline 236 and used a concentration range between 13 and 401 μg Hg/L. Lethal and developmental endpoints were examined at 24, 48, 72 and 96 hpf. Biochemical markers, including Acetylcholinesterase (AChE), Catalase (CAT), Glutathione Reductase (GR), and Glutathione-S-Transferase (GST) activities and, for the first time, the expression of the protein phosphatase 1 gamma (PP1γ) was assessed after 24, 48, 72 and 96 h of exposure to 10 and 100 μg Hg/L. The behavioral effects of a sublethal range of Hg (from 0.8 to 13 μg Hg/L) were assessed using an automated video tracking system at 120 hpf. Several developmental abnormalities on zebrafish embryos and larvae, including pericardial edema, spin and tail deformities and reduced rate of consumption of the yolk sac, were found after exposure to Hg (LC50 at 96 hpf of 139 μg Hg/L) with EC50 values for total malformations ranging from 22 to 264 μg Hg/L. After 96 hpf, no significant effects were observed in the CAT and GR activities. However, an increase in the GST activity in a concentration and time-dependent manner was found, denoting possible stress-related adaptation of zebrafish embryos to deleterious effects of Hg exposure. The AchE activity showed a response pattern in line with the behavioral responses. At the lowest concentration tested, no significant effects were found for the AChE activity, whereas a decrease in AChE activity was observed at 100 μg Hg/L, suggesting that exposure to Hg induced neurotoxic effects in zebrafish embryos which in turn may explain the lack of equilibrium found in this study (EC50 at 96 hpf of 83 μg Hg/L). Moreover, a decrease in the PP1γ expression was found after 96 h of exposure to 10 and 100 μg Hg/L. Thus, we suggest that Hg may be an inhibitor of PP1γ in zebrafish embryos-larvae and thus, along with the alterations in the enzymatic activity of GST, explain some of the developmental malformations observed, as well as the lack of equilibrium. Hence, in this study, we propose the use of PP1 expression, in combination with apical and biochemical endpoints, as a precursor for assessing Hg's toxic mechanism on embryonic development.

Initiation and duration of folic acid supplementation in preventing congenital malformations

BackgroundFolic acid (FA) supplementation is associated with a lower risk of the neural tube and heart defects and is recommended for women of childbearing age. Although there are detailed recommendations, differences in the initiation time and duration of FA supplementation remain poorly studied.MethodsA multicentre prospective study of 17,713 women was conducted. The incidence of congenital malformations in women taking a recommended dosage (e.g. 0.4 or 0.8 mg/day) of FA was compared with that in women without supplementation. The predicted probability of malformations by the initiation time and duration of FA use was estimated to determine optimal options.ResultsPericonceptional FA supplementation was associated with a lower and insignificant risk of congenital malformations (1.59% vs. 2.37%; odds ratio [OR] 0.69; 95% confidence interval [CI]: 0.44–1.08), heart defects (3.8 vs. 8.0 per 1000 infants; OR, 0.47; 0.21–1.02), and neural tube defects (7.0 vs. 11.5 per 10,000 infants; OR, 0.64; 0.08–5.15). FA use after pregnancy provided greater protection against total malformations. Statistically significant associations were found in women who initiated FA supplementation in the first month of gestation (OR, 0.55; 95% CI: 0.33–0.91) and in those who supplemented for 1 to 2 months (OR, 0.59; 95% CI: 0.36–0.98). Similar results were found for heart defects. The optimal initiation time was 1.5 (optimal range: 1.1 to 1.9) months before pregnancy and a duration of 4.0 (3.7 to 4.4) months was reasonable to achieve the lowest risk of congenital malformations. Heart defect prevention required an earlier initiation (2.2 vs. 1.1 months before pregnancy) and a longer duration (4.7 vs. 3.7 months) than the prevention of other malformations.ConclusionsThe timely initiation of FA supplementation for gestation was associated with a decreased risk of congenital malformations, which was mainly attributed to its protection against heart defects. The initiation of FA supplementation 1.5 months before conception with a duration of 4 months is the preferred option for congenital malformation prevention.Trial registrationChictr.org.cn identifier: ChiCTR-SOC-17010976.

Correção Cirúrgica de Desvio Angular Metatársico em uma Potra Puro-sangue Utilizando Ostectomia em Cunha: um relato de caso

Abstract This study describes a corrective procedure for a metatarsal angular malformation using wedge ostectomy in a 41-day-old Thoroughbred filly. A 41.6 °angular malformation of the metatarsus was corrected with wedge ostectomy of the left hindlimb in association with a type II external fixator and methyl methacrylate resin. The animal was subjected to the surgical procedure under general anesthesia. The filly was placed in dorsal recumbency with the left hind limb free for manipulation. A bone saw was used for the wedge ostectomy, while the bars for external fixation were placed using a drill. A Kirschner pin and acrylic resin were then applied. Radiographic images obtained immediately after surgery confirmed a total malformation reduction of 32.6 (from 41.6° to 9°). With medication and a Robert Jones bandage, intensive postoperative care was instituted to ensure that the animal adapted properly to the linear external fixator without overloading the pins and connection bars. No postoperative complications were observed. On the day after surgery, the filly was standing and walking with no signs of pain. After 180 days, total osseous calcification was achieved, and the implants were removed during the second surgery. The filly demonstrated adequate locomotion, performed all the movements, and ran without any difficulty in the paddock. The remaining discrete angular deviations were partially corrected using hoof trimming.

Congenita ptosis; Frequency of congenital ptosis.

Objectives: To compare the frequency of congenital ptosis of Pakistani children with the children of the world. Study Design: Comparative study. Setting: Al Shifa Tertiary Eye Hospital Rawalpindi. Period: August 2013 to September 2016. Material & Methods: The record of admitted patients of all malformations of eyes was collected from pediatric eye department from age zero to 15 years. The data of each eye malformation was documented separately as per age and gender. The total numbers of eye malformations were arranged according to prevalence, age and gender. The data was analyzed separately for frequency, age and gender. All data was analyzed and arranged according to percentage of frequency, gender and age. The age data was analyzed making different age groups. Seven groups of age were made with 2 years difference between each group starting from age 0 to 15 years. The number of male and females were calculated in each age group. Total numbers of each malformation was calculated separately and percentage of each malformation calculated as per simple mathematical way to total malformations. Results: The data of five hundreds and fourteen congenital malformations of eyes was collected. The numbers of male patients were 289 and female patients were 225. As per percentage the males were 56.2% and females patients were 43.8 %. The patient age range was from age zero to fifteen years. The majority of the patients were between ages zero to two years. The total numbers of patients in this age group were 222. Out of 222 patients the males were 120 and females were 102. The total percentage of this age group was 43.2 % among all age groups. The most common frequency of eye malformation was cataract i.e. 116 (22.6 %) patients followed by lacrimal apparatus anomalies i.e. 103(20 %) and ptosis was 45 (8.8 %). Conclusion: The frequency of congenital ptosis is the third most common eye malformation among total eye malformations in this study.

Use of embedded Chelex chelating resin and sediment toxicity bioassays with medaka embryos to determine the bioavailability and toxicity of lead-contaminated sediment

The aquatic sediment acts as a reservoir for multiple sources of pollutants including toxic metals. Most analytical methods used for estimating the bioavailability of sediment heavy metals have not been biologically validated by correlation with an aquatic organism's response. A reliable whole-sediment contacting toxicity assay using vertebrate species is lacking and the exposure routes for sediment metals are unclear. This study established a novel bio-analytical approach involving the Chelex-100 resin detection system and sediment toxicity assessment with embryo-larval stages of medaka fish (Oryzias latipes) to evaluate the bioavailability and toxicity of lead (Pb) contamination in sediment to fish. Treated fish exposed to the Pb-spiked artificial sediment with whole-sediment exposure showed more dose-dependent toxic responses than those from pore- or overlying-water exposure extracted from the same sediment. The Chelex-100 resin-extractable Pb content was highly correlated with mortality, total malformation and Pb bioaccumulation in medaka embryos or hatchlings from Pb-spiked sediment at environmentally relevant concentrations. The environmental sediment with higher contents of clay or organic carbon showed lower potency of releasing Pb from sediment to overlying water, as compared to those observed with artificial sediment. Our results suggest that the bio-analytical method can be practically applied in situ to evaluate the adverse effect of heavy metal-contaminated sediment on the aquatic ecosystem.

Developmental toxicity induced by Cu(OH)2 nanopesticide in zebrafish embryos.

The current study evaluates the adverse effects of Cu(OH)2 nanopesticide (CNPE) on the early life stages of zebrafish (Danio rerio). The developmental toxicity was determined using different parameters such as mortality (including LC50 ), hatching, heart rates, malformations, and alteration of the gene expressions. Zebrafish embryos (4 hpf-hours postfertilization) were exposed to 1.0, 2.0, 4.0, 8.0, and 16.0 mg/l CNPE doses until 96 hpf. The 96 hours LC50 was recorded at 6.258 mg/l. Seventy-two hpf total malformation index values for 2.0, 4.0, and 8.0 mg/l CNPE doses were 4.3, 7.2 and 7.9, respectively. 1.0 mg/l CNPE is not toxic for the zebrafish embryos/larvae. 2.0 to 8.0 CNPE doses caused some abnormalities in embryos/larvae morphology, including lack of body parts, tail deformities, chorda deformity, bubbled head, scoliosis, lordosis, weak or non-pigmentation, decreased heart rate and larva length. 16.0 mg/l CNPE caused mortality in 72 hpf. The expression levels of seven immune system-related genes (il-1β, il-8, cebp, tlr4, hsp70, NF-kB, and mtf-1) were examined. The transcription level of il-1β, il-8, tlr4, hsp70, and NF-kB genes significantly increased in the CNPE exposure groups. While the expression of the mtf-1 gene considerably decreased, the cebp gene expression level did not change in the 4.0 and 8.0 mg/l CNPE doses. In conclusion, CNPE could induce developmental toxicity with malformations in embryos/larvae and alter the gene expression.

Incubation Temperature Effect on Qualitative and Quantitative Composition of Abnormalities and Mortality Rate in Embryogenesis of the Barents Sea Capelin Mallotus villosus (Osmeridae)

The development of capelin Mallotus villosus from insemination to total resorption of the yolk is investigated at 5 and 8°C by the assessment of development of each individual (3850 eggs obtained from 77 spawning pairs). In total, ten malformation types are defined in early development. The frequency of their occurrence and embryonic mortality rate during incubation is described. The total malformation number and mortality rate are significantly higher at 8°С than at 5°С. A distinct relationship between early development quality and age or body size of the females is not revealed. In the majority of trials, the progenies of large females are characterized by better quality, but these data are not always significant. The qualitative composition and malformation rate of capelin differ from those observed in Atlantic cod Gadus morhua using the same methods of investigation. In addition, the malformation and mortality rates during early development are substantially lower in capelin than in Atlantic cod.

Gestational Exposure to Benzodiazepines, 2: The Risk of Congenital Malformations Examined Through the Prism of Compatibility Intervals.

A recent meta-analysis of the major malformation risk after gestational exposure to benzodiazepines identified 8 prospective cohort studies with 5,195 exposed and 2,082,467 unexposed women. Benzodiazepine exposure was not associated with a statistically significant increase in the risk of total malformations (8 studies) or cardiac malformations (4 studies). The malformation risk was not significantly increased after specifically first trimester exposure, either (5 studies). However, there was a significant increase in risk associated with combined first trimester exposure to benzodiazepines and antidepressants (3 studies). The authors of the meta-analysis interpreted their findings based on the conventional P < .05 cutoff for statistical significance. However, when the confidence intervals (CIs) associated with the pooled odds ratios (ORs) were examined, it was evident that almost the entire range of values in the CIs for total malformations and for cardiac malformations was compatible with a population value for the OR that indicated increased risk (OR > 1.00). Importantly, the CI for the first trimester exposure analysis was somewhat better distributed around the null (OR = 1.00), suggesting a lower likelihood of increased risk. All ORs were very low to low in value, indicating a very small increase in the absolute risk. Besides explaining how CIs may be interpreted as compatibility intervals, this article reminds readers that associations identified even in meta-analyses of cohort studies do not indicate a causal effect because confounding by indication can never be ruled out in observational research designs. A reasonable conclusion, therefore, is that gestational exposure to benzodiazepines is a marker of risk for cardiac and total malformations, and, importantly, that first trimester exposure to benzodiazepines may not be associated with much increase in risk, if at all.

Neonatal health including congenital malformation risk of 1072 children born after vitrified embryo transfer.

Does vitrification of Day 3 and Day 5 embryos adversely affect birth outcomes of singletons and twins in comparison with peers born after fresh embryo transfer? Neonatal health parameters, including the prevalence of congenital malformations, in singletons and twins born after embryo vitrification are similar to or slightly better than after fresh embryo transfer. Although vitrification, rather than slow-freezing, of embryos is routine practice nowadays, convincing evidence regarding the safety for the offspring is sparse. Literature data comprise results from mostly small-sized studies or studies including only Day 3 or only Day 5 vitrified embryo transfers. Overall, better or comparable perinatal outcomes, in terms of higher birthweight and lower risk for small-for-gestational age or for low birthweight, have been reported for singletons born after vitrified embryo transfer compared with fresh embryo transfer. According to the single available study with sufficient sample size, the congenital malformation rate was found to be comparable after vitrified and fresh embryo transfers. Data were collected from 960 cycles after transfer of embryos vitrified on Day 3 (n = 457) or Day 5 (n = 503) and from 1644 cycles after fresh embryo transfer on Day 3 (n = 853) or Day 5 (n = 791), performed between 2008 and 2013 at the Centre for Reproductive Medicine of the university hospital UZ Brussel. Outcome measures were neonatal health in terms of birthweight, small-for-gestational age, prematurity rate, perinatal death and major/minor/total malformation rate. Perinatal health parameters of 11 stillborns and 1061 live borns (827 singletons and 234 twins) in the vitrified group and of 28 stillborns and 1838 live borns (1374 singletons and 464 twins) in the fresh embryo group are reported. Within 3 months after birth, children in the two study groups were assessed clinically with special attention to congenital malformations by a paediatrician blinded to the type of embryo transfer. Data were analysed by multiple linear and logistic regression, adjusted for treatment variables and maternal characteristics. Mothers to infants in the vitrified group were on average slightly older and more often suffering from pregnancy-related hypertensive disorders than mothers to infants in the fresh transfer group. Singletons born after vitrification showed a higher birthweight standard deviation score (SDS) (-0.4 versus -0.7; 95% confidence interval (CI): 0.0-0.3, P = 0.001) and a lower small-for-gestational age rate (AOR: 0.55; 95% CI: 0.34-0.90) in comparison with peers born after fresh embryo transfer. Preterm birth rate and perinatal death rate were comparable between the two groups (AOR: 0.91; 95% CI: 0.57-1.43 and AOR: 0.97; 95% CI: 0.40-2.36). In twins, neonatal outcomes including birthweight SDS, small-for-gestational age and prematurity rates were comparable in the vitrified and the fresh groups, when adjusted for confounders. Furthermore, the rate of major congenital malformations in live borns was comparable between the vitrified group and the fresh group, both in singletons (2.6 versus 2.8%; AOR: 0.91; 95% CI: 0.47-1.78) and in twins (2.4 versus 2.7%; AOR: 0.51; 95% CI: 0.05-5.72). Also, the total malformation rate in the vitrified group (3.4%; 95% CI: 2.4-4.8) did not differ from the rate in the fresh embryo group (3.9%; 95% CI: 3.1-5.0). The embryonic stage at vitrification or fresh transfer (cleavage-stage embryo or blastocyst) did not influence the birth characteristics or malformation rate. The main limitation of this study is the rather small twin group. Therefore, the outcome results for twins should be interpreted cautiously. This study provides evidence that transfer of vitrified Day 3 and Day 5 embryos does not adversely affect the neonatal health of the offspring in comparison with transfer of fresh embryos. Furthermore, neonatal outcomes were not different after transfer of vitrified blastocysts compared with transfer of vitrified cleavage-stage embryos. Educational grants for establishing and organizing the data collection have come from IBSA, Ferring, Organon, Shering-Plough and Merck. Merck Belgium funded the data collection for outcomes after vitrification between 2012 and 2015. All co-authors, except M.B., declared no conflict of interest. M.B. has received consultancy fees from Organon, Serono Symposia and Merck.

Stages and directions of psychological support to families caring for children with severe congenital malformations

Nowadays the medical and social issues associated with hereditary and congenital pathologies, and first of all, congenital malformations and chromosomal diseases are very relevant. For a decade they have been assuming ever greater medical, social and economic importance. Long-term and complicated care of patients with congenital malformations, medical correction of the defects, pedagogical intervention as well as psychological support to social adaptation require a long time and much of finances, material and technical resources. Surgical pathologies in neonates and namely congenital malformations causing most perinatal and infant deaths are in the focus of current neonatology. The prevalence of congenital malformations of gastrointestinal tract and anterior abdominal wall ranges from 13 to 26, 4 cases per 10000 live-born neonates, and diaphragmatocele is registered in 2.5 – 5.0 cases per 10000 of livebirths. These pathologies account for 29.1% of the total congenial malformations ranking the 3 rd position. The most effective and reliable technique to spot congenital malformations of a fetus is ultrasound scanning. The optimal timing for detecting congenital malformations of gastrointestinal tract is 12 – 23 weeks of gestation. In addition to purely medical aspects of the diagnosis of birth defects which may be corrected surgically, there is another, but not less important psychological aspect. There is no doubt about the necessity of psychological support to parents and family members during the prenatal period (from diagnosis to birth), at all stages of surgical correction and postoperative care, regardless of their length. When any congenital complications have been detected the medical and psychological follow-up should be aimed to establish the most accurate diagnosis by experts, to provide detailed informing parents about the diagnosis and all up-to-date methods of treatment of this disease, to afford them ground for consulting with experts experienced in the treatment of certain pathologies. There are several behaviour types demonstrated by parents who become aware of their child’s diagnosis of congenital malformation: compensated families fully accepting a child and its defect (23%); families showing unacceptance of the child with birth defects, family with snail syndrome (26%), dysadvantaged families or families with low or unsatisfactory socio-cultural status (26%). Implementation of a certain type of behaviour by the family starts from the moment of diagnosis. Psychological correction in these families is multiple-aspect, complex and based on a number of principles, compliance with which ensures effective result of the influence: the principles of humane and empathic attitude, the unity of diagnosis and correction, harmonization of family relations, the encouragement of positive thinking towards the child, providing person-centred assistance, creating an inner positive plan for actions, the coordination in the activity of the medical staff, psychological groups and families. Psychological correction is performed in three main vectors (directions): a psychological study of the problem, psychological counselling, and providing psychological and educational support to family members. Prenatal diagnosis of congenital malformations shocks parents, causes psychological discomfort, in some cases mental and emotional stress with elements of emotional unacceptance of the child. The reaction of parents to the news that their child has birth defect is characterized by the staging that includes: phase of shock, phase of reactivity, adaptive phase, the phase of orientation. Since the baby has been born, the major stressful factor is staying in the atmosphere of uncertainty, expectations of surgical treatment and its results. Psychological supporter aimed to develop positive thinking, stipulates the following objectives: to correct unconstructive parental behaviour, to develop parents’ awareness of the problem of their child, to improve family relationship. Next, goals psychological rehabilitation of families where a child is brought up with the defect, the reconstruction of the hierarchy of values in life parents of a sick child, parents exercise personal growth in the process of interaction with your child. Further, the goals of psychological rehabilitation of families caring children with congenital malformation is the reconstruction of parents’ hierarchy of values in life, and the stimulation of personal growth in the process of interaction with the child.

Chronic effects of environmentally-relevant concentrations of lead in Pelophylax nigromaculata tadpoles: Threshold dose and adverse effects

Lead (Pb) is a common heavy metal in the natural environment, but its concentration has been increasing alongside widespread industrial and agricultural development in China. The dark-spotted frog Pelophylax (formerly Rana) nigromaculata (Anura: Ranidae) is distributed across East Asia and inhabits anthropogenic habitats such as farmland. Here, P. nigromaculata tadpoles (Gosner stage 19–46) were exposed to Pb at different concentrations (0, 40, 80, 160, 320, 640 and 1280µg/L) and Pb-induced survival, metamorphosis time, development, malformations, mobility and gonad structure were monitored. The results showed that above the threshold concentration of Pb, adverse effects were obvious. As the concentration of Pb increased, the adverse effects on different traits followed different patterns: the effects on hindlimb length, survival rate, metamorphosis rate, total malformation rate, swimming speed and jumping speed largely exhibited a linear pattern; the effects on snout–vent length, body mass and forelimb length largely exhibited a bimodal pattern. Sex ratio and gonadal histology were not affected by Pb, suggesting that Pb is not strongly estrogenic in P. nigromaculata.

Successful excision of a pontomesencephalic cavernoma through anterior subtemporal route without mapping: Anatomical landmarks as a road map

Successful excision of a pontomesencephalic cavernoma through anterior subtemporal route without mapping: Anatomical landmarks as a road map

Effect of certain plant oils on some biological and biochemical aspects on the cotton leaf worm Spodoptera littoralis.

The latent effects of three plant oils; namely thyme , bitter and neem on certain biological and biochemical parameters of the 6 th instar larvae of Spodoptera littoralis treated as 2 nd and 4 th instars with the LC50 values of these oils. The results showed that the 2 nd instars were more susceptible to all the tested oils than the 4 th instar larvae. All treatments recorded significantly difference in all durations for both 2 nd and 4 th instars. Also, highly significant prolongation in pupal duration and the most prolonged oil was occurred by thyme on the 2 nd instars, while bitter was more effective on the 4 th instar larvae. Longevity of both sexes was reduced specially for male moths. No significant reduction was noticed on female longevity as a result of pretreated 2 nd but significant reduction was recorded for pretreated 4 th instars especially with thyme. Highly significant reduction in pupal weight produced from the treated 2 nd and 4 th larval instars with the three products. All oils caused deformations with various degrees for larvae, pupae and adults resulted from the treated 2 nd and 4 th instar larvae. The highest deformations exist in larval-pupa intermediate by thyme as pretreated 2 nd and 4 th of the larval instars. Bitter gave the highest pupal deformation as the pretreated 4 th instars followed by neem for the 2 nd and 4 th instars. Generally, all oils exhibited higher effect on the males. Total malformation during all generation recorded its maximum value with neem treatment as 2 nd instars then thyme was the most effective in both 4 th and 2 nd of the larval instars followed by bitter. Highly significant stimulation in chitinase and α- & β -esterases activity was recorded with all treatments and the most effective one caused by thyme followed by bitter. Highly significant inhibition in protease activity and acetylcholine esterase (AChE) was attained by (bitter & thyme) and (neem & thyme) oils, respectively. While high significant stimulation was recorded in protease and non significant stimulation of AChE was noticed by neem and bitter, respectively.

Incidence and Variants of Posterior Arch Defects of the Atlas Vertebra

In order to describe the incidence and existing variants of congenital anomalies of the atlas vertebrae in a Caucasian population, we examined 1069 CT scans of the upper cervical spine. We found 41 cases with altered atlas vertebrae, representing 3.8% of all analyzed patients. With 83% of all found anomalies, the predominant type is characterized by a small dorsal cleft (3.2% of all patients). Rare varieties feature unilateral or bilateral dorsal arch defects, combined anterior and posterior clefts (0.2% of all patients) or total erratic atlas vertebra malformation (0.1% of all patients). Atlas arch defects are found nearly 4% at the time. Most anomalies affect the posterior arch, whereas the anterior arch or both are rarely affected. Totally irregular C1 vertebrae are extremely infrequent.

Neonatal outcome and congenital malformations in children born after ICSI with testicular or epididymal sperm: a controlled national cohort study

Does neonatal outcome including congenital malformations in children born after ICSI with epididymal and testicular sperm [testicular sperm extraction (TESE)/percutaneous epididymal sperm aspiration (PESA)/testicular sperm aspiration (TESA) (TPT)] differ from neonatal outcome in children born after ICSI with ejaculated sperm, IVF and natural conception (NC)? Children born after TPT have similar neonatal outcome, including total malformation rates, as have children born after ICSI and IVF with ejaculated sperm. Testing for variance over the four groups may indicate smaller differences in specific malformation rates with TPT as the highest risk group. Regarding neonatal outcome as well as congenital malformations in children born after TPT, studies are few, with limited sample size, heterogeneous and often performed without relevant control groups. Population-based cohort study including all Danish children born after TPT and fresh embryo transfer in Denmark from 1995 to 2009. Children born after transfer of frozen-thawed embryos were excluded. Control groups of children conceived by ICSI with ejaculated sperm, IVF and NC were identified by cross-linkage of the Danish IVF Register, Medical Birth Register (MBR) and National Hospital Discharge Register (HDR). The study group consisted of 466 children born after TPT, while the control groups consisted of 8967 (ICSI with ejaculated sperm), 17 592 (IVF) and 63 854 (NC) children. Neonatal outcomes and congenital malformations were analysed for singletons and twins separately. Risk estimates for low birthweight (LBW, <2500 g) and preterm birth (PTB, <37 gestational weeks) were adjusted for maternal age, parity, child gender and year of childbirth. The study group was identified from the Danish national database on children born after TPT. Control groups were obtained from the IVF register and the MBR. All information included in the study was retrieved from the national registers. Considering singletons and twins as one group, the sex ratio (♂/♀) was significantly lower for children born after TPT (0.89) compared with conventional IVF (1.11; P = 0.017) but did not differ significantly when compared with ICSI with ejaculated sperm (0.94) and NC (1.05). The mean birthweight (BW) for singletons did not differ significantly between groups when including only first-born children. The mean gestational age (GA) in the TPT singletons (279 ± 12 days) was significantly higher compared with IVF (276 ± 18 days; P = 0.02), but similar to ICSI with ejaculated sperm and NC singletons when including only first-born children (277 ± 16 days and 279 ± 14 days, respectively). Rate of stillbirths, perinatal and neonatal mortality in the group of TPT singletons did not differ significantly from any of the control groups. Comparable results were found for the TPT twin group, except for perinatal mortality, which was significantly lower in the TPT group compared with naturally conceived twins. The adjusted risk of LBW was significantly higher for TPT versus NC singletons [adjusted odds ratio (AOR) = 0.67 (0.48-0.93)]; however AOR for PTB was similar in the two groups. Regarding twins, similar adjusted risks were observed for PTB and LBW between the TPT and all three control groups. Significantly more Caesarean sections were performed after IVF (27.3% for singletons) and ICSI (25.1% for singletons) with ejaculated sperm compared with the TPT group (16.4% for singletons). The total rate of congenital malformations in the TPT group was 7.7% and did not differ significantly from any of the control groups. However, singleton TPT boys showed an increased rate of cardiac malformations (3.6%) compared with singleton boys after IVF (1.4%; P = 0.04) and NC (1.1%; P = 0.02). Considering the level of male infertility as a continuum over the four groups, tests for variance in the rate of cardiac malformations in singleton boys, and undescended testicles for singleton as well as twin boys were each significantly increased from NC to IVF to ICSI to TPT (P < 0.001). The rate of hypospadias showed the same pattern, but the TPT group did not differ significantly compared with the control groups. One of the limitations is that the TPT group could not be classified according to testicular or epididymal sperm, as these data were not available in the IVF register. Another limitation is that registry-based studies are encumbered with the risk of reporting or coding errors or missing data due to insufficient coding. However, the quality of data on congenital malformations in HDR has, in other studies, been validated and found acceptable for epidemiological research, and furthermore, recordings on study and control groups are performed similarly. Accumulating data show that TPT treatment is equally safe as conventional ICSI and IVF treatment and as NC with regard to neonatal outcome including congenital malformation. This study is supported by Laboratory of Reproductive Biology, Scientific Unit, Horsens Hospital. No competing interests declared.

The teratogenic effects of sediments from the Yangtze Estuary and adjacent bay, China, on frog embryos

In recent decades, the Yangtze (Changjiang) Estuary has suffered great environmental changes due to intensive human activities in the Yangtze River basin. This study assessed the ecotoxicity of sediments from the Yangtze Estuary with the frog embryo teratogenesis assay—Xenopus (FETAX). The results showed that the sediment extracts induced multiple malformations in embryos of Xenopus tropicalis. In the embryos treated with nearshore extracts, abnormal eyes, narrow fins and hypopigmentation were the dominant phenotypes, followed by enlarged proctodeums. In these embryos, the percentages of total malformations were greater than 50 % at five sampling sites and less than 40 % at the other seven sites. However, in the embryos treated with the offshore extracts, elongation of the proctodeums was the dominant phenotype, followed by bent tails. The percentages of total malformations ranged from 25 to 45 %. Comparatively, the embryos treated with extracts from the nearshore zone showed a greater variety of phenotypes than those treated with extracts from the offshore zone. These results suggest that the sediments showed high teratogenicity to the amphibian embryos. The results also indicate that the teratogencity to X. tropicalis embryos is a useful indicator of the pollution of sediments.

Abnormal elongation of the lower jaw in juvenile Japanese flounder: combined effects of a rotifer diet enriched with Nannochloropsis preserved by various methods and parentage

To elucidate possible causes of skeletal malformations in larval Japanese flounder, we reared larvae fed rotifers enriched with three types of preserved Nannochloropsis (fresh, refrigerated, and frozen). The incidence of malformations at 50 days post hatch ranged from 14.5 to 38.5 % within the three experimental groups, and elongation of the lower jaw (LJ) was the most frequently observed malformation, ranging from 68 to 89 % of total malformations. We also investigated larval parentages using microsatellite markers. Parentage analysis of the fresh Nannochloropsis group showed that one sire and a pair generated significant numbers of LJ-elongated individuals. In the refrigerated Nannochloropsis group, one dam and two sires generated significant numbers of LJ-elongated individuals. In the frozen Nannochloropsis group, no broodstocks or pairs generated significant numbers of LJ-elongated individuals. Our results suggest that LJ elongation in Japanese flounder likely results from the application of different types of preserved Nannochloropsis during rotifer feeding stage. However, there is also some level of genetic influence associated with this deformity.

Acute effects of Fe 2O 3, TiO 2, ZnO and CuO nanomaterials on Xenopus laevis

Metal oxide nanomaterials have exhibited toxicity to a variety of aquatic organisms, especially microbes and invertebrates. To date, few studies have evaluated the toxicity of metal oxide nanomaterials on aquatic vertebrates. Therefore, this study examined effects of ZnO, TiO 2, Fe 2O 3, and CuO nanomaterials (20–100 nm) on amphibians utilizing the Frog Embryo Teratogenesis Assay Xenopus (FETAX) protocol, a 96 h exposure with daily solution exchanges. Nanomaterials were dispersed in reconstituted moderately hard test medium. These exposures did not increase mortality in static renewal exposures containing up to 1000 mg L −1 for TiO 2, Fe 2O 3, CuO, and ZnO, but did induce developmental abnormalities. Gastrointestinal, spinal, and other abnormalities were observed in CuO and ZnO nanomaterial exposures at concentrations as low as 3.16 mg L −1 (ZnO). An EC 50 of 10.3 mg L −1 ZnO was observed for total malformations. The minimum concentration to inhibit growth of tadpoles exposed to CuO or ZnO nanomaterials was 10 mg L −1. The results indicate that select nanomaterials can negatively affect amphibians during development. Evaluation of nanomaterial exposure on vertebrate organisms are imperative to responsible production and introduction of nanomaterials in everyday products to ensure human and environmental safety.

Mango Malformation: I. Toxin Production Associated with Fusarium Pathogens

Eight Fusarium species i.e. F. subglutinans, F. solani, F. oxyspoum, F. sterilihyphosum, F. proliferatum, F. monili-forme, F. avena and F. chlamydspore isolated from mango malformed disease were tested for their ability to cause mango malformation disease and their production of moniliformin and total fumonisins (FB1 + FB2) using HPLC. A evaluated for moniliformin production, seven isolates were toxin producers, the production levels ranging from 0.51 to 8.90 µg/ml. The higher levels were produced by Fusarium subglutinans (8.51 µg/ml). Moderate concentrations of moniliformin was produced by F.moniliforme (6.90 µg/ml), F. oxysporum (6.30 µg/ml), F. proliferatum (4.10 μg/ml) and F. sterilihyphosum (1.10 μg/ml). Separation and identification of Fumonisin that was isolated from the pathogen- causing disease are made by (HPLC). A evaluated for total fumonisin production (FB1 + FB2), seven isolates were toxin producers, the production levels ranging from 0.10 to 8.30 µg/ml. The higher levels were produced by F. monili-forme (8.30 µg/ml. Moderate concentrations of fumonisin was produced by F .proliferatum (0.64 µg/ml) and F. subglutinans (0.50 µg/ml). Strong positive correlations between moniliformin and total fumonisins (FB1 + FB2) activities and malformation disease incidence by F. subglutinans, F. solani, F. oxyspoum, F. sterilihyphosum, F. proliferatum was observed.

Teratogenic effects of tetrabromobisphenol A on Xenopus tropicalis embryos

Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant and a known thyroid disruptor. We reported exposing Xenopus tropicalis embryos (NF10) to 0.01, 0.1 or 1 mg/L of TBBPA with or without 70 µg/L triiodothyronine (T 3). Compared with the controls, 1 mg/L of TBBPA significantly reduced the body length of embryos after 24, 36 and 48 h of exposure. Embryos treated with TBBPA showed multiple malformations, including: abnormal eyes, skin hypopigmentation, enlarged proctodaeum, narrow fins and pericardial edemas. The effect of abnormal eyes manifested itself in the loss of pigmentation, reduction in size, or absence of external eyes. The degree of eye malformations was quantified with the index of eye malformations (IEM) with 0 being normal and 3 being severe. In the 1 mg/L TBBPA treatment groups, the incidence of total malformations (ITM) was 68–93%, and IEM was 0.8–0.9. T 3 showed no teratogenic effects on embryos, but it significantly enhanced TBBPA-induced teratogenic effects. In the T 3 + 1 mg/L TBBPA treatment groups, ITM was 91–99%, and IEM was 1.8–1.9. Histological observations showed that the retinas were generally smaller, and the lenses were underdeveloped or even absent. These results indicate that TBBPA at relatively high concentration has teratogenic effects on X. tropicalis embryos. The results also suggest that thyroid hormone signaling might be involved in TBBPA induced-teratogenicity.