Novel insights into early life stages of finned octopods (Octopoda: Cirrata)

The finned or dumbo octopods (Octopoda: Cirrata) constitute a cephalopod (Mollusca: Cephalopoda) taxon almost exclusively comprising organisms that inhabit the deep-sea. This renders studying the general biology of adult cirrate specimens a difficult task, but even more so when it comes to gathering knowledge on their early life stages. During his comprehensive research exploits on cephalopod development, the late Sigurd von Boletzky (1942–2020) also sought to shed light on specific adaptations that eggs and embryos of these enigmatic deep-sea octopods might show. Based on his seminal work, the present article sets out to provide additional data on a broad range of early cirrate life stages, including egg capsules without any obvious embryonic stage as well as those containing embryos. These previously unreported specimens obtained from museum collections were analyzed using conventional morphological as well as modern three-dimensional imaging techniques. The present overview includes specimens from four of the five oceans, i.e., the Atlantic, Pacific, Indian, and Southern Ocean. Based on information on cirrate ovum, chorion, and egg capsule size as well as shape gathered from the literature, an attempt is made here for the first time to identify von Boletzky's as well as the specimens introduced here down to at least genus level. The combined data provide novel insights into early life stages of finned octopods, aiming to continue von Boletzky's legacy with regard to developmental research on a still largely enigmatic taxon of extant deep-sea cephalopods.

Developing Inside a Layer of Germs—A Potential Role for Multiciliated Surface Cells in Vertebrate Embryos

This paper reviews current research on the microbial life that surrounds vertebrate embryos. Several clades are believed to develop inside sterile—or near-sterile—embryonic microhabitats, while others thrive within a veritable zoo of microbial life. The occurrence of embryo-associated microbes in some groups, but not others, is an under-appreciated transition (possibly transitions) in vertebrate evolution. A lack of comparable studies makes it currently impossible to correlate embryo-associated microbiomes with other aspects of vertebrate evolution. However, there are embryonic features that should instruct a more targeted survey. This paper concludes with a hypothesis for the role of multiciliated surface cells in amphibian and some fish embryos, which may contribute to managing embryo-associated microbial consortia. These cells are known to exist in some species that harbor in ovo microbes or have relatively porous egg capsules, although most have not been assayed for embryo-associated microbiota. Whether the currents generated within these extraembryonic microhabitats contribute to culturing consistent microbial communities remains to be seen.

Snails associated with the coral-killing sponge Terpios hoshinota in Okinawa Island, Japan

Terpios hoshinota is a thin encrusting sponge that overgrows live scleractinian corals and it is linked to coral loss in many reefs. However, our knowledge of the species associated with this sponge species is poor. During a periodical survey of T. hoshinota in 2020, we found tiny snails crawling on the sponge in the subtropical waters around Okinawa Island, Japan. We observed egg capsules inside the sponge tissue and veliger larvae released from the egg capsules. Molecular analyses of both the snails and veliger larvae (cytochrome oxidase I, COI) showed that they were identical and belonged to Joculator sp. (family Cerithiopsidae). There was no direct observation of predation on the sponge by this snail; however, to the best of our knowledge, this is the first report on a close association between a snail and the sponge T. hoshinota.

Micro-CT protocols for scanning and 3D analysis of Hexaplex trunculus during its different life stages

Micro-computed tomography (micro-CT) is a high-resolution 3D-imaging technique which is now increasingly applied in biological studies focusing on taxonomy and functional morphology. The creation of virtual representations of specimens can increase availability of otherwise underexploited and inaccessible samples. The 3D model dataset can be also further processed through volume rendering and morphometric analysis. The success of micro-CT as a visualisation technique depends on several methodological manipulations, including the use of contrast enhancing staining agents, filters, scanning mediums, containers, exposure time and frame averaging. The aim of this study was to standardise a series of micro-CT scanning and 3D analysis protocols for a marine gastropod species, Hexaplex trunculus. The analytical protocols have followed all the developmental stages of this gastropod, from egg capsules and embryos to juveniles and adults.

MicroCT protocols for scanning egg capsules of Hexaplex trunculus v1

Micro-computed tomography (micro-CT) is a high-resolution 3D-imaging technique which is now increasingly applied in biological studies focusing on taxonomy and functional morphology. The creation of virtual representations of specimens can increase availability of otherwise underexploited and inaccessible samples. This protocol aims to standardise micro-CT scanning procedures for the egg capsules of the marine gastropod species Hexaplex trunculus.

Distribution of Gonipterus Species and Their Egg Parasitoids in Australia: Implications for Biological Control

Gonipterus species are pests of Eucalyptus plantations worldwide. The egg parasitoid wasp Anaphes nitens is used in many countries for the biological control of Gonipterus spp. Recent taxonomic studies have shown that the three invasive Gonipterus spp., which were previously considered as G. scutellatus, form part of a cryptic species complex. These taxonomic changes have implications for the biological control of Gonipterus spp. The aims of this study were to understand the species composition and distribution of Gonipterus spp. and their egg parasitoids in Australia. Gonipterus spp. adults and egg capsules were collected in south-eastern Australia and Tasmania. Adult Gonipterus were identified using morphology and DNA barcoding. Parasitoids were reared from Gonipterus egg capsules and identified. Thirteen Gonipterus species were collected: twelve species were found on the Australian mainland and one species in Tasmania. These included three described species, four previously recognized but undescribed species, two undescribed species and four unidentified species. Five egg parasitoid species that attack Gonipterus spp. were identified. Anaphes nitens, Centrodora damoni and Euderus sp. were identified on the Australian mainland and A. tasmaniae and A. inexpectatus were identified in Tasmania. The results from this study will contribute to the improvement of Gonipterus biological control in the future.

Field Monitoring of Tritia mutabilis (Linnaeus, 1758) Egg Capsule Deposition and Intracapsular Embryonic Patterns Using Artificial Substrates and Machine Learning-Based Approaches

Tritia mutabilis (Linnaeus, 1758) represents a marine gastropod species of ecological and economical importance especially in coastal areas of the central Adriatic Sea (Italy). T. mutabilis encloses its fertilized eggs within capsules which play a protective role in the embryo defense against adverse environmental factors. Egg capsules are attached to any hard substrates and the availability of adequate substrates for oviposition represents therefore a major determinant of reproductive output of this species. The aim of the present study was to provide the first comprehensive monitoring of T. mutabilis egg capsule deposition and intracapsular embryonic patterns using specific artificial substrates and innovative analytical approaches. Square-based pyramid structures were placed within an area subjected to small-scale fishing activity along the Italian coastal waters of the central Adriatic Sea and monitored from March to June 2019. In addition, a machine learning-based approach was developed in order to speed up and automate time-consuming counting procedures of egg capsules attached on each pyramid. Overall, egg capsules were deposited on the almost totality of the artificial substrates reaching values of the surface coverage per site ranging from 31 to 97%. These findings lead to a quantitative estimation of egg number per site included between 27,060 and 62,940 capsules. Our results demonstrate that using these artificial substrates together with monitoring of both egg capsule deposition patterns and intracapsular developmental stages may be useful to improve the T. mutabilis stock management.

A new piece in the puzzle for the riverine slugs of the Acochlidiidae (Gastropoda: Panpulmonata: Acochlidimorpha) helps tracing steps of their freshwater invasion

Gastropods (slugs and snails) are prominent and species-rich faunal elements in marine and terrestrial habitats of the tropics. While several clades of snails inhabit freshwater systems, slugs are extremely rare in freshwater: only the centimeter-sized Acochlidiidae, with currently three genera, contain more than one species and live in the lower reaches of island streams in an area comprising Eastern Indonesia, Fiji, and Palau. Where known, the species of this unique group are specialized predators of other amphidromous snails’ egg capsules (Neritidae) and their reproductive biology and adaptations to life in freshwater are complex. Acochlidiidae are thus of special interest for evolutionary biology and ecology. We here describe a new genus of unusually bluish-green acochlidiid to date known only from a single locality on the island of Ambon, Indonesia. Previous molecular data found this charismatic species to link slender Strubellia slugs with broad and flattened Acochlidium and Palliohedyle. We establish Wallacellia siputbiru n. gen. n. sp., the “blue slug” in Bahasa Indonesia, by using scanning electron microscopy of cuticular elements, light microscopy of serial semithin histological sections of the soft body, and 3D reconstruction of all organ systems based on these sections. Special structures of this seemingly rare endemic species include the enlarged kidney and the huge copulatory organ. Our data now clarify that, in the invasion of freshwater habitats in Acochlidiidae, sexual selection (the anterior three-part copulatory organ) preceded ecological selection (posterior flattened habitus with branched or multiplied internal organs).