A simple sexing test for elephant species and its application to faecal DNA.

We developed a novel real-time PCR assay for rapid sexing in all three elephant species, which amplifies small fragments of the orthologous sexual chromosome zinc finger protein genes ZFX/ZFY (65 bp). This assay is a simple, inexpensive and reliable tool that is suitable for non-invasive DNA samples and can be incorporated into larger SNP panels for individual identification and population genetic studies.

Sturgeon conservation genomics: SNP discovery and validation using RAD sequencing.

Caviar-producing sturgeons belonging to the genus Acipenser are considered to be one of the most endangered species groups in the world. Continued overfishing in spite of increasing legislation, zero catch quotas and extensive aquaculture production have led to the collapse of wild stocks across Europe and Asia. The evolutionary relationships among Adriatic, Russian, Persian and Siberian sturgeons are complex because of past introgression events and remain poorly understood. Conservation management, traceability and enforcement suffer a lack of appropriate DNA markers for the genetic identification of sturgeon at the species, population and individual level. This study employed RAD sequencing to discover and characterize single nucleotide polymorphism (SNP) DNA markers for use in sturgeon conservation in these four tetraploid species over three biological levels, using a single sequencing lane. Four population meta-samples and eight individual samples from one family were barcoded separately before sequencing. Analysis of 14.4 Gb of paired-end RAD data focused on the identification of SNPs in the paired-end contig, with subsequent in silico and empirical validation of candidate markers. Thousands of putatively informative markers were identified including, for the first time, SNPs that show population-wide differentiation between Russian and Persian sturgeons, representing an important advance in our ability to manage these cryptic species. The results highlight the challenges of genotyping-by-sequencing in polyploid taxa, while establishing the potential genetic resources for developing a new range of caviar traceability and enforcement tools.

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 June 2011-31 July 2011.

This article documents the addition of 112 microsatellite marker loci and 24 pairs of single nucleotide polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Agelaius phoeniceus, Austrolittorina cincta, Circus cyaneus, Circus macrourus, Circus pygargus, Cryptocoryne × purpurea Ridl. nothovar. purpurea, Mya arenaria, Patagioenas squamosa, Prochilodus mariae, Scylla serrata and Scytalopus speluncae. These loci were cross-tested on the following species: Cryptocoryne × purpurea nothovar. purpurea, Cryptocoryne affinis, Cryptocoryne ciliata, Cryptocoryne cordata var. cordata, Cryptocoryne elliptica, Cryptocoryne griffithii, Cryptocoryne minima, Cryptocoryne nurii and Cryptocoryne schulzei. This article also documents the addition of 24 sequencing primer pairs and 24 allele-specific primers or probes for Aphis glycines.

Reproducibility of sonographic assessment of fetal liver length in diabetic pregnancies.

OBJECTIVES: Assessing fetal liver size might be useful in monitoring the effectiveness of the treatment of diabetes in pregnancy. We aimed to assess the reproducibility of fetal liver-length measurement in pregnant women with diabetes. METHODS: From 3 April 2006 to 5 December 2006, we assessed intraobserver and interobserver variation of fetal liver-length measurements on 55 sonograms in 44 pregnant women with diabetes, 34 of whom had gestational diabetes. The mean maternal age was 33 years, the mean maternal weight was 92 kg and the mean body mass index (BMI) was 33.7 kg/m(-2). The effect of covariates BMI, gestational age and maternal age on the reproducibility of fetal liver length was assessed by calculating intraobserver SD ratios. We compared liver length with abdominal circumference and gestational age. Nine of 12 sonographers scanned, on average, six women (range, 3-12) as the first sonographer, and all 12 sonographers scanned, on average, four women (range, 1-10) as the second sonographer. The data were analyzed using a hierarchical linear model. RESULTS: Measurement of fetal liver length was reproducible. The intraobserver SD was 3.06 (95% CI, 2.68-3.59) mm; the interobserver SD was 2.17 (95% CI, 0.59-4.83) mm; the intraobserver correlation was 0.77 (95% CI, 0.63-0.87), and the interobserver correlation was 0.84 (95% CI, 0.51-0.99). The covariate effects were minimal, the SD for a 1-unit increase in the covariate was 1.06 for gestational age, 0.98 for BMI, and 0.97 for maternal age. CONCLUSIONS: Measurement of fetal liver length in the diabetic pregnancy is reproducible. It is worthy of further investigation as a parameter for monitoring the effectiveness of treatment of the diabetic pregnancy.

Mitochondrial DNA sequences from dried snake venom: a DNA barcoding approach to the identification of venom samples.

Outdated nomenclature and incorrect taxonomic characterisation of snake venoms in the current toxinological literature have serious implications for the replicability of results from snake venom toxin research. The situation has not improved, despite attempts to supply toxinologists with regular updates on snake systematics. Here, we demonstrate the successful extraction of DNA, and subsequent sequencing of the mitochondrial 12S gene, from dried snake venoms. This approach offers a new and potentially straightforward method for accurate species identification. Mitochondrial DNA (mtDNA) sequences isolated from snake venom can be used to clarify or validate snake species identification through comparison against existing sequences in the GenBank database, and through phylogenetic analyses with other sequences. Pooled venoms can also be screened a priori for the presence of multiple species, and the species names on the labels of commercial venoms verified. Moreover, if the species from which the venom sample has been taken is known, and the specimen is available as a voucher, the mtDNA sequence of the haplotype isolated from that species venom sample could serve as a sequence standard (or 'DNA barcode') for that species. Our new method of DNA barcoding venoms ensures the identification of venoms even after future taxonomic changes.

Is conservative management of prenatal and neonatal ovarian cysts justifiable?

OBJECTIVES: To evaluate conservative management of fetal and neonatal ovarian cysts. METHODS: We followed the progress of 11 fetal and 2 neonatal ovarian cysts. RESULTS: Ten fetal cysts were simple at recognition, but 8 silently became complex by birth, with ovarian loss in 6/11 cases (54%). In 2 cases where large simple cysts presented for the first time at birth, serious complications occurred, resulting in the death of 1 newborn. CONCLUSIONS: If a fetal ovarian cyst is already complex before birth it can be managed conservatively, albeit with loss of the ipsilateral ovary in the majority of cases. As simple fetal ovarian cysts frequently become complex before birth, intervention may be justifiable to preserve the ovary. But any intervention also has the potential for complications, and may be ineffective. Intervention also requires accurate differentiation of ovarian cysts from other abdominal cysts.

Congenitally corrected transposition of the great arteries: clues for prenatal diagnosis.

Congenitally corrected transposition of the great arteries (ccTGA) is an uncommon cardiac defect characterized by the atria connecting with the anatomically discordant ventricles and the ventricles connecting with discordant and transposed great arteries. Parallel vessels are evident in corrected TGA, but as this sign is also present in complete TGA, a heart anomaly requiring major cardiac surgery in the postnatal period, it is important to differentiate between the entities prenatally. Most cases of ccTGA have associated anomalies but isolated forms or those with a mild associated cardiac anomaly are infrequently detected prenatally. We report on three cases detected between 21 and 25 weeks' gestation on screening ultrasound with associated mild findings. One fetus had an isolated ventricular septal defect (VSD) first detected at 34 weeks. The child developed heart block at 4 years of age. The second case was associated with a small VSD, a tiny pulmonary trunk and a persistent right umbilical vein. After birth, mild pulmonary stenosis was found as an additional cardiac finding at 4 months of age. The third fetus had no additional cardiac anomalies prenatally, but after birth a bicuspid aortic valve was detected. The first case needed pacemaker implantation but the other two children required no cardiac surgery. Two of the cases were referred because abnormal vessel anatomy was detected on screening ultrasound. As prenatal detection of TGA is becoming a more frequent occurrence, this paper aims to present clues aiding in the prenatal diagnosis of atrioventricular and ventriculoarterial discordance, especially in its differentiation from complete transposition. These details are crucial for counseling and perinatal management.

Enteric duplications presenting as antenatally detected abdominal cysts: is delayed resection appropriate?

PURPOSE: The aim of this study was to evaluate delayed elective resection of antenatally detected enteric duplication cysts. METHODS: A retrospective casenote study of intraabdominal cysts detected antenatally between January 1991 and January 2002 found 37 fetuses with cysts. Twelve were enteric duplications. Two were duodenal, 1 was an 85-cm tubular jejunoileal duplication, and 9 were ileocecal. Asymptomatic cysts were followed with serial ultrasound scars and resected electively over 14 months. RESULTS: Three neonates had small bowel obstruction demanding laparotomy: 1 of the 2 infants with duodenal duplication cysts, 1 infant with an ileocecal duplication, and the infant with the tubular duplication. One with an ileocecal duplication became symptomatic at 2 months and underwent a laparotomy. Seven had their duplications resected electively between 6 weeks and 14 months, and the other is still being followed. Four of the 7 asymptomatic duplications electively resected contained gastric mucosa. CONCLUSIONS: Intraabdominal enteric duplication cysts are increasingly likely to be detected antenatally. The majority are likely to remain asymptomatic for several months at least, after which a resection can be planned. The prevalence of gastric mucosa suggests that they should not be left indefinitely. Laparoscopically assisted resection of ileocecal duplications is safe and effective.

Three cross-sectional planes for fetal color Doppler echocardiography.

Routine use of color Doppler during every fetal cardiac examination remains controversial. Many examiners still believe that color should be reserved for cases of suspected congenital heart defect (CHD). In our opinion, color Doppler should be applied in every cardiac scan due to the increase in speed and accuracy that it allows. The purpose of this review is to first explain how color Doppler presets can be optimized and, second, to propose the use of three cross-sectional planes to simplify color Doppler fetal echocardiography: the four-chamber (4CV), five-chamber (5CV) and three-vessel (3VV) views. A practical approach to the detection of CHD with these planes is presented, with typical findings and possible abnormalities evident during systole and diastole. The diastolic pattern on the 4CV is characterized by two equal color stripes. Connection ('H'-sign) or size inequality of the two stripes, or a unilateral color stripe, are important abnormal findings. In systole valve regurgitation should be excluded. In the 5CV, turbulent flow, ventricular septal defect or an overriding aorta ('Y'-sign) can be detected. In the 3VV the aorta and pulmonary trunk should be of nearly equal size and demonstrate antegrade flow. Abnormal findings encountered include absence of one vessel, discrepant size of the vessels, retrograde flow in one of the vessels, or the 'U'-sign, where the trachea is enclosed between both vessels, suggesting right-sided aortic arch. In summary, we propose that color Doppler examination utilizing these three planes alone is sufficient to obtain adequate information for the detection of most common CHD.