Topographical mapping of sympathetic postganglionic innervation of mouse heart v1

This protocol describes the process of mapping the topographical organization of tyrosine hydroxylase immune reactive sympathetic postganglionic axons and terminals in the mouse heart. Hearts were removed and separated as whole mounts, then scanned using confocal or zeiss microscopy

Mapping CGRP-IR innervation of male mice stomach with Neurolucida 360 v1

This protocol describes the process of using Neurolucida 360 software to map the topographical organization of Calcitonin gene related peptide – immunoreactive axons and terminals in the muscular layer of mice stomach. Stomachs were removed, layers were separated and gone under immunohistochemistry as whole mounts, then scanned using confocal microscopy.

Evolution and development of the bird chondrocranium

Background Birds exhibit an enormous diversity in adult skull shape (disparity), while their embryonic chondrocrania are considered to be conserved across species. However, there may be chondrocranial features that are diagnostic for bird clades or for Aves as a whole. We synthesized and analyzed information on the sequence of chondrification of 23 elements in ten bird species and five outgroups. Moreover, we critically considered the developmental morphology of the chondrocrania of 21 bird species and examined whether the diversity in adult skull shape is reflected in the development of the embryonic skull, and whether there are group-specific developmental patterns. Results We found that chondrocranial morphology is largely uniform in its major features, with some variation in the presence or absence of fenestrae and other parts. In kiwis (Apteryx), the unique morphology of the bony skull in the orbito-nasal region is reflected in its chondrocranial anatomy. Finally, differences in morphology and chondrification sequence may distinguish between different Palaeognathae and Neognathae and between the Galloanserae and Neoaves. The sequence of chondrification is largely conserved in birds, but with some variation in most regions. The peri- and prechordal areas in the base of the chondrocranium are largely conserved. In contrast to the outgroups, chondrification in birds starts in the acrochordal cartilage and the basicranial fenestra is formed secondarily. Further differences concern the orbital region, including early chondrification of the pila antotica and the late formation of the planum supraseptale. Conclusion Synthesizing information on chondrocranial development confronts terminological issues and a lack of comparable methods used (e.g., different staining; whole-mounts versus histology). These issues were taken into consideration when assessing differences across species. The summary of works on avian chondrocranial development, covered more than a century, and a comparison of the chondrification sequence among birds could be conducted. Future studies could test the hypothesis that chondrocranial disparity in Aves, in terms of the shape and proportion of individual elements, could be as large as adult skull disparity, despite conserved developmental patterns and the richness of forms in other (dermal) portions of the skull.

Five new species of cotylean flatworms (Platyhelminthes: Polycladida: Cotylea) from Oaxaca, southern Mexican Pacific

The flatworms of the suborder Cotylea live in diverse marine substrates such as rocks, seaweed and sand or in association with corals. In the Mexican Pacific, particularly on the coast of Oaxaca, knowledge about cotyleans is scarce, with only one record of the genus Pseudoceros. The aim of this work was to describe the cotylean species present in the area. A total of 48 specimens were collected, fixed, mounted in whole mounts or histological sagittal and frontal histological sections of the reproductive systems. These were examined for taxonomic identification, determining five new species of four genera: Boninia oaxaquensis sp. nov., Pericelis sigmeri sp. nov., Pericelis nazahui sp. nov., Pseudoceros bicuti sp. nov. and Thysanozoon estacahuitensis sp. nov. The present study establishes the first record of the genus Boninia and the family Boniniidae for the Tropical Eastern Pacific. It also confirms the presence of the genus Pseudoceros on the coast of Oaxaca, and break the disjunct distribution of this genus (from Gulf of California to Gulf of Tehuantepec), as well as the genus Pericelis (from Canada to Galapagos Islands) and Thysanozoon (from Chile to Gulf of California).

Stains for entomological microtechnique: simple stains for whole mounts and dissection

Slide mounted entomological specimens often require the aid of contrast techniques to improve the clarity of morphological characteristics. Methods can involve the use of techniques such as Phase contrast, Dark field or differential interference contrast microscopy (DIC), however where an entomologist may only have access to simple brightfield microscopy chemical staining of the specimen may be used to improve contrast. For whole mounts of entomological specimens, a single stain, occasionally two, are often used, in comparison to histological sections that often employ multiple stains in complex protocols. A number of authors have proposed different stains and staining methods for a number of insect groups with few considering the long term qualities of the stain, it has previously been shown that aniline dyes are prone to fading in Canada Balsam mounts, and that some stains fade even when protected from sunlight. This paper aims to summarise the knowledge of stains used for entomological specimens and provide details on the archival qualities.

Astacopsidrilus hibernicus sp. nov. (Phreodrilidae, Oligochaeta, Annelida) from Irish peatlands

The discovery of a large and flourishing population of Phreodrilidae in terrestrial peatlands in northwest Ireland was surprising on two counts: these oligochaete worms are usually aquatic and most of the species occur in the Southern Hemisphere. The phreodrilids were discovered in a project that targeted Enchytraeidae, therefore methods adapted to the investigation of enchytraeids could be applied, including the study of living animals and properly fixed whole mounts. DNA sequencing was also performed. All worms identified here belong to one species, new to science, and placed in the genus Astacopsidrilus, because of the ventral position of the spermathecal pores and the opening of the female funnels inside the spermathecal vestibule. Astacopsidrilus hibernicus sp. nov. is mainly distinguished by thick segmental cushions of epidermal gland cells on the dorsal side of the posterior body half. Male sexual organs and spermathecae are comparatively small and without the often-observed bizarre modifications common in species of this family. DNA sequencing yielded a fragment of the 16S rRNA gene. This is the first description of a phreodrilid species from Europe; the few previous recordings of this family in Ireland and the United Kingdom had been left unidentified.

Trypanorhynch cestodes parasitizing Mugil liza (Mugiliformes: Mugilidae) commercialized in the state of Rio de Janeiro, Brazil

The aim of the present study was to identify, through morphological and morphometric analyses, the species of trypanorhynch cestodes found as plerocerci in the intestinal serosa of Mugil liza and to determine their parasitic indices. One hundred and fifty specimens of this mullet collected off the coast of the state of Rio de Janeiro were necropsied and the trypanorhynch cestodes found were fixed and preserved as whole mounts for morphological analysis. The plerocerci were identified as Callitetrarhynchus gracilis and Pterobothrium crassicole, both with a prevalence of 0.67%, an intensity of 1 and abundance of 0.0067, in single infections. This is the first record of a trypanorhynch cestode parasitizing M. liza in Brazil. Although the parasitic indexes were low, from a hygienic-sanitary perspective the plerocerci of these species are visible to the naked eye, and thus can disgust consumers and make marketing the fish unfeasible.