The phylogeny of the Casque-headed Treefrogs (Hylidae: Hylinae: Lophyohylini).

The South American and West Indian Casque-headed Treefrogs (Hylidae: Hylinae: Lophyohylini) include 85 species. These are notably diverse in morphology (e.g. disparate levels of cranial hyperossification) and life history (e.g. different reproductive modes, chemical defences), have a wide distribution, and occupy habitats from the tropical rainforests to semiarid scrubland. In this paper, we present a phylogenetic analysis of this hylid tribe based on sequence fragments of up to five mitochondrial (12S, 16S, ND1, COI, Cytb) and six nuclear genes (POMC, RAG-1, RHOD, SIAH, TNS3, TYR). We included most of its species (> 96%), in addition to a number of new species. Our results indicate: (i) the paraphyly of Trachycephalus with respect to Aparasphenodon venezolanus; (ii) the nonmonophyly of Aparasphenodon, with Argenteohyla siemersi, Corythomantis galeata and Nyctimantis rugiceps nested within it, and Ap. venezolanus nested within Trachycephalus; (iii) the polyphyly of Corythomantis; (iv) the nonmonophyly of the recognized species groups of Phyllodytes; and (v) a pervasive low support for the deep relationships among the major clades of Lophyohylini, including C. greeningi and the monotypic genera Itapotihyla and Phytotriades. To remedy the nonmonophyly of Aparasphenodon, Corythomantis, and Trachycephalus, we redefined Nyctimantis to include Aparasphenodon (with the exception of Ap. venezolanus, which we transferred to Trachycephalus), Argenteohyla, and C. galeata. Additionally, our results indicate the need for taxonomic work in the following clades: (i) Trachycephalus dibernardoi and Tr. imitatrix; (ii) Tr. atlas, Tr. mambaiensis and Tr. nigromaculatus; and (iii) Phyllodytes. On the basis of our phylogenetic results, we analyzed the evolution of skull hyperossification and reproductive biology, with emphasis on the multiple independent origins of phytotelm breeding, in the context of Anura. We also analyzed the inter-related aspects of chemical defences, venom delivery, phragmotic behaviour, co-ossification, and prevention of evaporative water loss.

Distribution modeling of Amblyomma rotundatum and Amblyomma dissimile in Brazil: estimates of environmental suitability.

The number of reports of tick parasitism in amphibians and reptiles has increased over the past few years, including new host and location records for Amblyomma rotundatum and Amblyomma dissimile. However, knowledge of the geographical distribution remains incomplete, and in many regions of Brazil, the presence of these vectors has not been investigated. Several candidate models were built using a correlative maximum entropy approach, and best-fitting models were selected based on statistical significance, predictive ability, and complexity based on current climatic trends and future projected climate changes. Final models showed a good ability to discriminate A. rotundatum and A. dissimile current and future potential distributions. The entire country had higher predicted suitability for A. rotundatum while A. dissimile was mainly restricted to the Amazon and Pantanal biomes. A. rotundatum is a species with enormous potential for dissemination in the next decades, potentially through the legal and illegal transport of reptiles and amphibians. The proposed model is useful for targeting surveillance efforts increasing the efficiency and accuracy of future ecological research and tick management efforts.

Methods of Granulocyte Isolation from Human Blood and Labeling with Multimodal Superparamagnetic Iron Oxide Nanoparticles.

This in vitro study aimed to find the best method of granulocyte isolation for subsequentlabeling with multimodal nanoparticles (magnetic and fluorescent properties) to enable detectionby optical and magnetic resonance imaging (MRI) techniques. The granulocytes were obtained fromvenous blood samples from 12 healthy volunteers. To achieve high purity and yield, four differentmethods of granulocyte isolation were evaluated. The isolated granulocytes were labeled withmultimodal superparamagnetic iron oxide nanoparticles (M-SPIONs) coated with dextran, and theiron load was evaluated qualitatively and quantitatively by MRI, near-infrared fluorescence (NIRF)and inductively coupled plasma mass spectrometry (ICP-MS). The best method of granulocyteisolation was Percoll with Ficoll, which showed 95.92% purity and 94% viability. After labeling withM-SPIONs, the granulocytes showed 98.0% purity with a yield of 3.5 × 106 cells/mL and more than98.6% viability. The iron-loading value in the labeled granulocytes, as obtained by MRI, was 6.40 ±0.18 pg/cell. Similar values were found with the ICP-MS and NIRF imaging techniques. Therefore,our study shows that it is possible to isolate granulocytes with high purity and yield and labelingwith M-SPIONs provides a high internalized iron load and low toxicity to cells. Therefore, these MSPION-labeled granulocytes could be a promising candidate for future use ininflammation/infection detection by optical and MRI techniques.

Phylogenetic systematics of egg-brooding frogs (Anura: Hemiphractidae) and the evolution of direct development.

Egg-brooding frogs (Hemiphractidae) are a group of 105 currently recognized Neotropical species, with a remarkable diversity of developmental modes, from direct development to free-living and exotrophic tadpoles. Females carry their eggs on the back and embryos have unique bell-shaped gills. We inferred the evolutionary relationships of these frogs and used the resulting phylogeny to review their taxonomy and test hypotheses on the evolution of developmental modes and bell-shaped gills. Our inferences relied on a total evidence parsimony analysis of DNA sequences of up to 20 mitochondrial and nuclear genes (analyzed under tree-alignment), and 51 phenotypic characters sampled for 83% of currently valid hemiphractid species. Our analyses rendered a well-resolved phylogeny, with both Hemiphractidae (sister of Athesphatanura) and its six recognized genera being monophyletic. We also inferred novel intergeneric relationships [((Cryptobatrachus, Flectonotus), (Stefania, (Fritziana, (Hemiphractus, Gastrotheca))))], the non-monophyly of all species groups previously proposed within Gastrotheca and Stefania, and the existence of several putative new species within Fritziana and Hemiphractus. Contrary to previous hypotheses, our results support the most recent common ancestor of hemiphractids as a direct-developer. Free-living aquatic tadpoles apparently evolved from direct-developing ancestors three to eight times. Embryos of the sister taxa Cryptobatrachus and Flectonotus share a pair of single gills derived from branchial arch I, while embryos of the clade including the other four genera have two pairs of gills derived from branchial arches I and II respectively. Furthermore, in Gastrotheca the fusion of the two pairs of gills is a putative synapomorphy. We propose a revised taxonomy concordant with our optimal topologies.