Effects of Ocean Acidification on Molting, Oxidative Stress, and Gut Microbiota in Juvenile Horseshoe Crab Tachypleus tridentatus

Anthropogenic elevation of atmospheric carbon dioxide (CO2) drives global-scale ocean acidification (OA), which has aroused widespread concern for marine ecosystem health. The tri-spine horseshoe crab (HSC) Tachypleus tridentatus has been facing the threat of population depletion for decades, and the effects of OA on the physiology and microbiology of its early life stage are unclear. In this study, the 1st instar HSC larvae were exposed to acidified seawater (pH 7.3, pH 8.1 as control) for 28 days to determine the effects of OA on their growth, molting, oxidative stress, and gut microbiota. Results showed that there were no significant differences in growth index and molting rate between OA group and control group, but the chitinase activity, β-NAGase activity, and ecdysone content in OA group were significantly lower than those of the control group. Compared to the control group, reactive oxygen species (ROS) and malondialdehyde (MDA) contents in OA group were significantly increased at the end of the experiment. Superoxide dismutase (SOD), catalase (CAT), and alkaline phosphatase (AKP) activities increased first and then decreased, glutathione peroxidase (GPX) decreased first and then increased, and GST activity changed little during the experiment. According to the result of 16S rRNA sequencing of gut microbiota, microbial-mediated functions predicted by PICRUSt showed that “Hematopoietic cell lineage,” “Endocytosis,” “Staphylococcus aureus infection,” and “Shigellosis” pathways significantly increased in OA group. The above results indicate that OA had no significant effect on growth index and molting rate but interfered with the activity of chitinolytic enzymes and ecdysone expression of juvenile horseshoe crabs, and caused oxidative stress. In addition, OA had adverse effects on the immune defense function and intestinal health. The present study reveals the potential threat of OA to T. tridentatus population and lays a foundation for the further study of the physiological adaptation mechanism of juvenile horseshoe crabs to environmental change.

The Owadów-Brzezinki geoeducation area at Sławno

The geoeducation area (called also “Owadów-Brzezinki Geopark”) located in the north-western margin of the Holy Cross Mountains (Tomaszów Syncline) at Sławno community (Łódź Voivodeship), was established in June 2019, in close vicinity of the Owadów-Brzezinki quarry. This locality is one of the most important palaeontological sites described recently in Poland. The area consists of the exhibition pavilion, educational routs and panoramic viewing platform, which is located along the edge of the quarry. The palaeontological exhibition shows the unique Late Jurassic fossils of marine and terrestrial organisms, many of them new to science, that have been excavated in the quarry during the last eight years. Among the most important fossils are: ammonites, lobster-like decapod crustaceans, horseshoe crabs, actinopterygian fish, a cryptodiran turtle, ichthyosaurs, as well as a small terrestrial crocodyliform, pterosaurs and insects. In addition to the original fossils, the exhibition presents life-size reconstructions of animals, that inhabited the local seas and islands during the Late Jurassic. The palaeontological sites of Owadów-Brzezinki is referred to as a new “taphonomic window” of the Late Jurassic, providing insights about the evolution of life on Earth in the palaeogeographical and palaeoenvironmental context.

What Is an “Arachnid”? Consensus, Consilience, and Confirmation Bias in the Phylogenetics of Chelicerata

The basal phylogeny of Chelicerata is one of the opaquest parts of the animal Tree of Life, defying resolution despite application of thousands of loci and millions of sites. At the forefront of the debate over chelicerate relationships is the monophyly of Arachnida, which has been refuted by most analyses of molecular sequence data. A number of phylogenomic datasets have suggested that Xiphosura (horseshoe crabs) are derived arachnids, refuting the traditional understanding of arachnid monophyly. This result is regarded as controversial, not least by paleontologists and morphologists, due to the widespread perception that arachnid monophyly is unambiguously supported by morphological data. Moreover, some molecular datasets have been able to recover arachnid monophyly, galvanizing the belief that any result that challenges arachnid monophyly is artefactual. Here, we explore the problems of distinguishing phylogenetic signal from noise through a series of in silico experiments, focusing on datasets that have recently supported arachnid monophyly. We assess the claim that filtering by saturation rate is a valid criterion for recovering Arachnida. We demonstrate that neither saturation rate, nor the ability to assemble a molecular phylogenetic dataset supporting a given outcome with maximal nodal support, is a guarantor of phylogenetic accuracy. Separately, we review empirical morphological phylogenetic datasets to examine characters supporting Arachnida and the downstream implication of a single colonization of terrestrial habitats. We show that morphological support of arachnid monophyly is contingent upon a small number of ambiguous or incorrectly coded characters, most of these tautologically linked to adaptation to terrestrial habitats.

Assessing horseshoe crab avoidance response of flicker and continuous red-light LED: Gillnets bycatch mitigation

The horseshoe crab has the important roles as macrobenthos, but it is unfriendly for gillnets fisheries because it can damage the fishing net. The horseshoe crab is an ancient, rare, and protected animal so that the fishermen unable to utilize it, so the effort like mitigations need to be conducted like using red light LED. To avoid the learning behavior of horseshoe crab, so that the technology used is red light LED continuous then the red light led with flicker is used as a comparison. This study aims to determine the right type of red-light LED for horseshoe crab bycatch mitigation by the response pattern. The method used was a laboratory experiment. There are 20 adult horseshoe crabs used. The analysis used was descriptively comparative. The results showed that the red-light LED with flicker was most avoided by horseshoe crabs, which is 75%, while the fastest response of horseshoe crab in avoiding light was found in the light with a flicker effect of 22.97 seconds. Based on the horseshoe crab’s response, it can be concluded that the red LED light flicker is better than the red LED continuous as an alternative technology for the mitigation of horseshoe crab bycatch.

New records of cheilostome Bryozoa from the eastern coast of India encrusting on the exoskeleton of live horseshoe crabs of Indian Sundarbans

Bryozoans are common commensal on hard surfaces and cover slow-moving animals like molluscans, sea turtles, brachyuran crabs, and horseshoe crabs. A total of six species of bryozoans belonging to four genus under three families of order Cheilostomatida were recorded encrusting on the carapaces of horseshoe crabs collected from Indian Sundarbans along the east coast of India and two among them, viz., Biflustra savartii (Audouin, 1826) and Sinoflustra arabianensis (Menon & Nair, 1975) are reported for the first time. Additionally, Jellyella tuberculata (Bosc, 1802) is reported for the first time from West Bengal coastal waters, previously known only from the Odisha coast of India. Both male and female horseshoe crabs were found to have been encrusted with bryozoan mats, although adequately not known about the life stages of their encrustation.

DNA Metabarcoding Revealed Interspecific Dietary Difference and Prey Selectivity in Juvenile Horseshoe Crabs Carcinoscorpius rotundicauda and Tachypleus tridentatus From Hong Kong

Horseshoe crabs provide important ecological services including bioturbation and linking of food web in the shallow waters, but their populations are declining globally, leading to major concerns on conservation of these iconic animals. Baseline information of horseshoe crab ecology, such as their trophic role and food source, is pre-requisite for habitat protection plan and captive restocking program. Trophic ecology of Asian horseshoe crabs is relatively poorly understood and previous studies on their juveniles have suggested that they are selective feeders rather than opportunistic generalists. This study demonstrates a non-invasive approach, using DNA metabarcoding analyses of the nuclear 18S rRNA gene on fecal samples to assess the dietary compositions of Carcinoscorpius rotundicauda and Tachypleus tridentatus juveniles to (1) determine their dietary compositions and trophic roles in their ecosystem, (2) determine any prey selectivity, and (3) distinguish the interspecific dietary differences with potential implications on the habitat requirement and ecological partitioning between these two horseshoe crab species. The results based on relative read abundance (RRA) suggested that oligochaetes were the major prey items for both C. rotundicauda (41.6%) and T. tridentatus (32.4%). Bivalves and crustaceans were second major prey groups for C. rotundicauda (8.6 and 8.4%, respectively). Surprisingly, anthozoans contributed a considerable portion of T. tridentatus’s diet (22.8%), which has never been reported. Furthermore, the major prey groups identified in the fecal samples were not the dominant benthic organisms in the studied area as revealed by environmental DNA (eDNA) analyses on the sediment samples, implying that both species are selective feeders rather than dietary generalists. Significant differences observed in the dietary compositions of the two species might be partially due to the difference in habitat preference between the two species. This study provides new insights into the trophic ecology of the two Asian horseshoe crab species in the estuarine habitat and establishes a new framework for future detailed molecular dietary analyses on all developmental stages of horseshoe crabs around the world, which will allow us to evaluate the food sources needed for the survival of horseshoe crabs and facilitate future conservation strategies without killing the animals.

Widespread retention of ohnologs in key developmental gene families following whole genome duplication in arachnopulmonates

Whole genome duplications have occurred multiple times during animal evolution, including in lineages leading to vertebrates, teleosts, horseshoe crabs and arachnopulmonates. These dramatic events initially produce a wealth of new genetic material, generally followed by extensive gene loss. It appears, however, that developmental genes such as homeobox genes, signalling pathway components and microRNAs are frequently retained as duplicates (so called ohnologs) following whole-genome duplication. These not only provide the best evidence for whole-genome duplication, but an opportunity to study its evolutionary consequences. Although these genes are well studied in the context of vertebrate whole-genome duplication, similar comparisons across the extant arachnopulmonate orders are patchy. We sequenced embryonic transcriptomes from two spider species and two amblypygid species and surveyed three important gene families, Hox, Wnt and frizzled, across these and twelve existing transcriptomic and genomic resources for chelicerates. We report extensive retention of putative ohnologs, further supporting the ancestral arachnopulmonate whole-genome duplication. We also found evidence of consistent evolutionary trajectories in Hox and Wnt gene repertoires across three of the six arachnopulmonate orders, with inter-order variation in the retention of specific paralogs. We identified variation between major clades in spiders and are better able to reconstruct the chronology of gene duplications and losses in spiders, amblypygids, and scorpions. These insights shed light on the evolution of the developmental toolkit in arachnopulmonates, highlight the importance of the comparative approach within lineages, and provide substantial new transcriptomic data for future study.