Description of the oocysts of three new species of 'Eimeria' (Apicomplexa: Eimeriidae) from Iguanid lizards (Sauria: Iguanidae) of Central and South America

Three new species of Eimeria are described from iguanid lizards of Central and South America. The oocysts of each species have no micropyles or residua and the sporocysts lack Stieda bodies, but all have a sporocyst residuum. Eimeria sanctaluciae n.sp. was found in the St. Lucia tree lizard, Anolis luciae, collected from the Maria Islands, Lesser Antilles. The oocysts are spherical to subspherical, averaging 17.3 x 16.5 microns, with a single layered colourless wall; about 60% contain polar granules. The sporocysts are ellipsoidal and average 7.7 x 5.5 microns. Eimeria liolaemi n.sp. was recovered from the blue-gold swift, Liolaemus taenius, from Chile. The oocysts are spherical to subspherical, measuring 21 x 20.1 microns with a single-layered colourless wall. The sporocysts are subspherical and average 7.4 x 6.8 microns. Eimeria caesicia n.sp. is described from the Brazilian collared iguanid, Tropidurus torquatus. The oocysts measure 27.4 x 23.7 microns, are spherical to subspherical, with a bilayered wall, the outer surface of which appears pale blue in colour, the thin, inner wall appearing brown, when viewed by direct light under the optical microscope. The sporocysts are subspherical and average 9.4 x 7.2 microns. Unnamed polysporocystid oocysts with dizoic sporocysts are reported from the faeces of the lesser St. Vincent tree lizard, Anolis trinitatis and the possibility of spurious parasitism briefly discussed. In addition, oocysts of an unnamed Isospora sp. with a smooth oocyst wall which closely resembles I. reui were recovered from A. trinitatis

A novel likely pathogenic variant in the RUNX1 gene as the cause of congenital thrombocytopenia

Abstract Introduction: Heterozygous pathogenic and likely pathogenic sequence variants in the RUNX1 (Runt-related Transcription Factor 1) gene are a common genetic cause of decreased platelet count and/or platelet dysfunction and an increased risk of developing myelodysplasia and acute myeloid leukemia. The majority of causative variants are substitutions, which rarely occur de novo. The aim of this case report is to present a patient with congenital thrombocytopenia caused by a deletion variant in exon 9 in the RUNX1 gene. Case report: A one-month-old male infant was admitted to the Clinical Hospital Center Rijeka because of anemia and thrombocytopenia verified in the course of an acute viral infection. During follow-up, he occasionally had petechiae and ecchymoses on the lower extremities after mild trauma, with no other symptoms. The patient had persistent slightly decreased values of platelets with normal morphology, but with pathological aggregation with adrenaline and adenosine diphosphate. Due to the unclear etiology of persistent mild thrombocytopenia, he was referred for genetic testing at the age of five. Genomic DNA was isolated from the patient’s peripheral blood and whole-exome sequencing was performed using the nextgeneration sequencing method. A heterozygous frameshift variant, c.1160delG (NM_001754.4), was identified in exon 9. The variant is classified as likely pathogenic. Conclusion: To the best of our knowledge, the heterozygous variant c.1160delG in the RUNX1 gene was first described in our patient. Although pathogenic variants in the RUNX1 genes are very rare, persistently low platelet counts of unclear etiology should raise suspicion of an underlying genetic disorder

Shotgun metagenomic sequencing in culture negative microbial keratitis

Purpose To evaluate the microbiota of culture negative Corneal Impression Membrane (CIM) microbial keratitis samples with the use of shotgun metagenomics analysis. Methods DNA of microbial keratitis samples were collected with CIM and extracted using the MasterPure (TM) Complete DNA and RNA Purification Kit (Epicentre). DNA was fragmented by sonication into fragments of 300 to 400 base pairs (bp) using Bioruptor (R) (Diagenode, Belgium) and then used as a template for library preparation. DNA libraries were sequenced on Illumina (R) HiSeq2500. The resulting reads were quality controlled, trimmed and mapped against the human reference genome. The unmapped reads were taxonomically classified using the Kraken software. Results 18 microbial keratitis samples were included in the study. Brevundimonas diminuta was found in 5 samples while 6 samples showed the presence of viral infections. Cutibacterium acnes, Staphylococcus aureus, Moraxella lacunata and Pseudomonas alcaligenes were also identified as the presumed putative cause of the infection in 7 samples. Conclusions Shotgun sequencing can be used as a diagnostic tool in microbial keratitis samples. This diagnostic method expands the available tests to diagnose eye infections and could be clinically significant in culture negative samples