INVERTEBRATE BIOLOGY

The life-history fitness of F hybrids of the microcrustacean and (Crustacea, Anomopoda)
Moy I, Green M, Pham TP, Luu D and Xu S
Negative interaction between alleles that arise independently in diverging populations (i.e., Dobzhansky-Muller incompatibilities) can cause reduction of fitness in their hybrids. However, heterosis in hybrids can emerge if hybridization breaks down detrimental epistatic interaction within parental lineages. In this study, we examined the life-history fitness of the inter-specific Fs of two recently diverged microcrustacean species and as well as intra-specific crosses of . We identified heterosis in two out of five life-history traits in the inter-specific Fs. According to theories that heterosis can transiently emerge in early speciation, the observation of heterosis in these life-history traits suggests that there are no major genetic incompatibilities between these two species affecting these traits and that and are at an early stage of speciation.
Providing interactive and field laboratories while teaching university marine biology classes in an era of COVID-19
Cowles DL and Onthank KL
An entirely online upper-division university marine invertebrates course modeled after a field experience-intensive course that also provided interaction with live animals and research experience was offered at Rosario Beach Marine Laboratory in the summer of 2020. We describe online methods we used for providing field experiences to students participating online, as well as a workstation and interactive method for identification and detailed anatomical examination of live macroinvertebrates with students. Students were also involved as active participants in a field research project. Nearly all of the equipment involved is inexpensive or readily available in most university biology laboratories or classrooms.
Vascular architecture in the bacteriogenic light organ of (Cephalopoda: Sepiolidae)
Patelunas AJ and Nishiguchi MK
Symbiosis between southern dumpling squid, (Cephalopoda: Sepiolidae), and its luminescent symbiont, the bacterium , provides an experimentally tractable system to examine interactions between the eukaryotic host and its bacterial partner. Luminescence emitted by the symbiotic bacteria provides light for the squid in a behavior termed "counter-illumination," which allows the squid to mask its shadow amidst downwelling moonlight. Although this association is beneficial, light generated from the bacteria requires large quantities of oxygen to maintain this energy-consuming reaction. Therefore, we examined the vascular network within the light organ of juveniles of with and without . Vessel type, diameter, and location of vessels were measured. Although differences between symbiotic and aposymbiotic squid demonstrated that the presence of does not significantly influence the extent of vascular branching at early stages of symbiotic development, these finding do provide an atlas of blood vessel distribution in the organ. Thus, these results provide a framework to understand how beneficial bacteria influence the development of a eukaryotic closed vascular network and provide insight to the evolutionary developmental dynamics that form during mutualistic interactions.
Carboniferous Onychophora from Montceau-les-Mines, France, and onychophoran terrestrialization
Garwood RJ, Edgecombe GD, Charbonnier S, Chabard D, Sotty D and Giribet G
The geological age of the onychophoran crown-group, and when the group came onto land, have been sources of debate. Although stem-group Onychophora have been identified from as early as the Cambrian, the sparse record of terrestrial taxa from before the Cretaceous is subject to contradictory interpretations. A Late Carboniferous species from the Mazon Creek biota of the USA, , originally interpreted as a crown-group onychophoran, has recently been allied to early Cambrian stem-group taxa. Here we describe a fossil species from the Late Carboniferous Montceau-les-Mines Lagerstätte, France, informally referred to as an onychophoran for more than 30 years. The onychophoran affinities of gen. nov., sp. nov. are indicated by the form of the trunk plicae and the shape and spacing of their papillae, details of antennal annuli, and the presence of putative slime papillae. The poor preservation of several key systematic characters for extant Onychophora, however, prohibits the precise placement of the Carboniferous fossil in the stem or crown of the two extant families, or the onychophoran stem-group as a whole. Nevertheless, is the most compelling candidate to date for a terrestrial Paleozoic onychophoran.
Structure of the male copulatory apparatus in (Platyhelminthes, Kalyptorhynchia)
Doe DA and Smith JP
Gnathorhynchidae is a diverse taxon of predatory eukalyptorhynch flatworms characterized by an armed proboscis. Their present taxonomy is not concordant with what we know of their phylogeny. Further progress in this area is hindered by a lack of information concerning their morphology. As recent studies have shown, a historical reliance on live observations for species descriptions has resulted in a number of errors and omissions. Here, we redescribe the anatomy of the male copulatory organ of using transmission-electron and confocal microscopy, correcting several errors in the original description. Furthermore, we use these results to update our understanding of the anatomy and evolution of male copulatory organs in Gnathorhynchidae and in Platyhelminthes more generally.
Tuning of shortening speed in coleoid cephalopod muscle: no evidence for tissue-specific muscle myosin heavy chain isoforms
Shaffer JF and Kier WM
The contractile protein myosin II is ubiquitous in muscle. It is widely accepted that animals express tissue-specific myosin isoforms that differ in amino acid sequence and ATPase activity in order to tune muscle contractile velocities. Recent studies, however, suggested that the squid might be an exception; members of this species do not express muscle-specific myosin isoforms, but instead alter sarcomeric ultrastructure to adjust contractile velocities. We investigated whether this alternative mechanism of tuning muscle contractile velocity is found in other coleoid cephalopods. We analyzed myosin heavy chain transcript sequences and expression profiles from muscular tissues of a cuttlefish, , and an octopus, , in order to determine if these cephalopods express tissue-specific myosin heavy chain isoforms. We identified transcripts of four and six different myosin heavy chain isoforms in and muscular tissues, respectively. Transcripts of all isoforms were expressed in all muscular tissues studied, and thus and do not appear to express tissue-specific muscle myosin isoforms. We also examined the sarcomeric ultrastructure in the transverse muscle fibers of the arms of and the arms and tentacles of using transmission electron microscopy and found that the fast contracting fibers of the prey capture tentacles of have shorter thick filaments than those found in the slower transverse muscle fibers of the arms of both species. It thus appears that coleoid cephalopods, including the cuttlefish and octopus, may use ultrastructural modifications rather than tissue-specific myosin isoforms to adjust contractile velocities.
Textures and traction: how tube-dwelling polychaetes get a leg up
Merz RA
By controlling the traction between its body and the tube wall, a tube-dwelling polychaete can move efficiently from one end of its tube to the other, brace its body during normal functions (e.g., ventilation and feeding), and anchor within its tube avoiding removal by predators. To examine the potential physical interaction between worms and the tubes they live in, scanning electron microscopy was used to reveal and quantify the morphology of worm bodies and the tubes they produce for species representing 13 families of tube-dwelling polychaetes. In the tubes of most species there were macroscopic or nearly macroscopic (∼10 μm-1 mm) bumps or ridges that protruded slightly into the lumen of the tube; these could provide purchase as a worm moves or anchors. At this scale (∼10 μm-1 mm), the surfaces of the chaetal heads that interact with the tube wall were typically small enough to fit within spaces between these bumps (created by the inward projection of exogenous materials incorporated into the tube wall) or ridges (made by secretions on the interior surface of the tube). At a finer scale (0.01-10 μm), there was a second overlap in size, usually between the dentition on the surfaces of chaetae that interact with the tube walls and the texture provided by the secreted strands or microscopic inclusions of the inner linings. These linings had a surprising diversity of micro-textures. The most common micro-texture was a "fabric" of secreted threads, but there were also orderly micro-ridges, wrinkles, and rugose surfaces provided by microorganisms incorporated into the inner tube lining. Understanding the fine structures of tubes in conjunction with the morphologies of the worms that build them gives insight into how tubes are constructed and how worms live within them.
The secret to a successful relationship: lasting chemistry between ascidians and their symbiotic bacteria
Schmidt EW
Bioactive secondary metabolites are common components of marine animals. In many cases, symbiotic bacteria, and not the animals themselves, synthesize the compounds. Among marine animals, ascidians are good models for understanding these symbioses. Ascidians often contain potently bioactive secondary metabolites as their major extractable components. Strong evidence shows that ~8% of the known secondary metabolites from ascidians are made by symbiotic bacteria, and indirect evidence implicates bacteria in the synthesis of many more. Far from being "secondary" to the animals, secondary metabolites are essential components of the interaction between host animals and their symbiotic bacteria. These interactions have complex underlying biology, but the chemistry is clearly ascidian-species specific. The chemical interactions are ancient in at least some cases, and they are widespread among ascidians. Ascidians maintain secondary metabolic symbioses with bacteria that are phylogenetically diverse, indicating a convergent solution to obtaining secondary metabolites and reinforcing the importance of secondary metabolism in animal survival.
Diagnostic PCR can be used to illuminate meiofaunal diets and trophic relationships
Maghsoud H, Weiss A, Smith JP, Litvaitis MK and Fegley SR
Analysis of the meiofaunal food web is hampered because few prey have features that persist long enough in a predator's digestive tract to allow identification to species. Hence, at least for platyhelminth predators, direct observations of prey preference are almost nonexistent, and where they occur, prey identification is often limited to phylum. Studies using an approach are rare because they are extremely time-consuming and are subject to the criticism that predators removed from their natural environment may exhibit altered behaviors. Although PCR-based approaches have achieved wide application in food-web analysis, their application to meiofaunal flatworms suffers from a number of limitations. Most importantly, the microscopic size of both the predator and prey does not allow for removal of prey material from the digestive tract of the predator, and thus the challenge is to amplify prey sequences in the presence of large quantities of predator sequence. Here, we report on the successful use of prey-taxon-specific primers in diagnostic PCR to identify, to species level, specific prey items of 13 species of meiofaunal flatworms. Extension of this method will allow, for the first time, the development of a species-level understanding of trophic interactions among the meiofauna.
The reproductive system of (Annelida, Siboglinidae): ovary structure, sperm ultrastructure, and fertilization mode
Katz S and Rouse GW
is a genus of siboglinid annelids in which the females live on dead vertebrate bones on the seafloor. These females have a posterior end that lies within the bone and contains the ovarian tissue, as well as the "roots" involved with bone degradation and nutrition. The males are microscopic and live as "harems" in the lumen of the gelatinous tube that surrounds the female trunk, well away from the ovary. Females are known to spawn fertilized primary oocytes, suggesting internal fertilization. However, little is known about sperm transfer, sperm storage, or the location of fertilization, and the morphology of the female reproductive system has not been described and compared with the reproductive systems of other siboglinids. A 3D-reconstruction of the ovisac of showed ovarian tissue with multiple lobes and mature oocytes stored in a "uterus" before being released through the single oviduct. The oviduct emerges as a gonopore on the trunk and travels along the trunk to finally open to the seawater as a thin cylindrical tube among the crown of palps. Light and transmission electron microscopy of mature sperm revealed elongate heads consisting of a nucleus with helical grooves occupied by mitochondria. In contrast to other Siboglinidae, sperm are not packaged into spermatophores or spermatozeugmata, and females lack a discrete region for sperm storage. Transmission electron microscopy and fluorescence microscopy allowed detection of sperm associated with ovarian tissue of the female ovisac of four different species. This provides the first evidence for the site of internal fertilization in . A heart body was found in the circulatory system, as seen in other siboglinids and some other annelids. The possible presence of nephridia in the anterior ovisac region was also documented. These morphological features provide new insights for comparing the regionalization of females in relation to other siboglinids.
Effects of food type, feeding frequency, and temperature on juvenile survival and growth of Marisa cornuarietis (Mollusca: Gastropoda)
Selck H, Aufderheide J, Pounds N, Staples C, Caspers N and Forbes V
The present experiments are part of a larger study designed to investigate the influence of husbandry parameters on the life history of the ramshorn snail, Marisa cornuarietis, in order to identify suitable husbandry conditions for maintaining multi-generation populations in the laboratory for use in ecotoxicological testing. In this paper we focus on the effects of a combination of food types and feeding frequencies (i.e., the frequency with which the snails were offered food) on juvenile growth and survival at different temperatures. Offspring produced in the laboratory by wild specimens of M. cornuarietis, from Puerto Rico, were used to test the effects of three types of food (lettuce, alginate with fish food, alginate with snail mix) fed at three frequencies (given ad libitum on 4/4, 2/4, or 1/4 d) on juvenile survival and growth. The 4-d feeding regimens were repeated four times, giving a total of 16 d for the experiments. The experiments were conducted at two temperatures (22 degrees and 25 degrees C) under a 12 h light:12 h dark photoperiod. Juvenile growth rates increased with increasing feeding frequency for all food types. The most rapid growth rates occurred in the high-frequency lettuce treatments and the slowest growth rates in the low-frequency lettuce and alginate with snail mix treatments. Juvenile snails grew faster at 25 degrees than at 22 degrees C, and mortality was about twice as high at the lower temperature. Growth rates were used to provide a rough estimate of time to maturity, which was determined to take about twice as long at 22 degrees than at 25 degrees C. The results showed that lettuce is the best food if supplied in abundance, but effects on growth are very dependent on feeding frequency and temperature. We conclude that 25 degrees C is a more appropriate temperature for maintaining populations than 22 degrees C, that lettuce provides a suitable food source, and that food should be supplied continuously for husbandry and toxicity testing of populations of M. cornuarietis.
Effects of husbandry parameters on the life-history traits of the apple snail, Marisa cornuarietis: effects of temperature, photoperiod, and population density
Aufderheide J, Warbritton R, Pounds N, File-Emperador S, Staples C, Caspers N and Forbes V
These experiments are part of a larger study designed to investigate the influence of husbandry parameters on the life history of the apple snail, Marisa cornuarietis. The overall objective of the program is to identify suitable husbandry conditions for maintaining multi-generation populations of this species in the laboratory for use in ecotoxicological testing. In this article, we focus on the effects of photoperiod, temperature, and population density on adult fecundity and juvenile growth. Increasing photoperiod from 12 to 16 h of light per day had no effect on adult fecundity or egg hatching and relatively minor effects on juvenile growth and development. Rearing snails at temperatures between 22 degrees C and 28 degrees C did not influence the rates of egg production or egg clutch size. However, the rates of growth and development (of eggs and juveniles) increased with increasing temperature in this range, and when temperatures were reduced to 22 degrees C egg-hatching success was impaired. Juvenile growth and development were more sensitive to rearing density than adult fecundity traits. On the basis of the present results, we conclude that rearing individuals of M. cornuarietis at a temperature of 25 degrees C, a photoperiod of 12L:12D, and a density of <0.8 snails L(-1) (with lower densities for juvenile snails) should provide favorable husbandry conditions for maintaining multi-generation populations of this species.