Yasuhisa Henmi

Biology of the Amphioxus, Branchiostoma belcheri in the Ariake Sea, Japan II. Reproduction

Abstract We investigated the reproduction of the amphioxus Branchiostoma belcheri in the southern Ariake Sea, Japan, from 1999 to 2002. Gonads were very small or almost absent from September to December and began to develop in January. Changes in the gonad length index indicated that spawning began in mid June, after the gonads had attained maximum size. Although most gametes were extruded during the first spawning, some remained in the gonads. Shrunken gonads were much smaller but contained gametes, and the second spawning occurred around 10 July. It was unclear whether all adults spawned twice, but a large proportion of individuals did. After the first spawning, both males and females lost 30% of their body weight. Most one-year-old individuals did not spawn, and the minimum size at maturity was ca. 20 mm. We found no termination of reproduction among very large individuals. The average number of gonads was significantly larger on the right side of the body (26.2 in males and 26.1 in females) than on the left side (24.0 in males and 23.5 in females). We found two hermaphrodites in a total of 11,184 specimens examined. Each had four or three ovaries among a total of 45 or 54 gonads.

Biology of the Amphioxus, Branchiostoma belcheri in the Ariake Sea, Japan I. Population Structure and Growth

Abstract We investigated the population structure and growth of the amphioxus Branchiostoma belcheri for four years in the southern Ariake Sea, Japan. We counted 62-66 myotomes and 251-310 dorsal fin-ray chambers, and these results support that this species is an intermediate form of B. belcheri and its subspecies B. belcheri tsingtauense. The ratio of females to males was 1:1.12. Males were more numerous than females among small individuals (< 40 mm body length), but we found no significant differences among large animals (≥ 50 mm body length). Spawning occurred from mid June to early July. Groups of newly settled young appeared from January to June of their second year. We observed a large fluctuation between years in the numbers of newly settled young. The estimated size of one-year-old individuals was 19.4 mm in body length; within the next 12 months, they reached 32.1 mm. Three- and four-year-old individuals measured 38.6 mm and 45.8 mm, respectively. Few grew beyond 60 mm; the largest specimen collected was a 64 mm male.

To court or not to court: reproductive decisions by male fiddler crabs in response to fluctuating food availability

For males, courting and foraging are often behavioral alternatives, which take time and consume energy. When males have a possibility of mating with receptive females, there may be a behavioral trade-off between courtship and feeding; the outcome of which may be affected by male physiological condition and food availability. Although many mathematical models and empirical studies suggest that the expression of male courtship signals are condition-dependent, decisions about courtship and mating strategies in relation to food availability have not attracted much attention. In this study, we tested whether daily changes in food availability affect males’ decisions about whether to court. We conducted experiments with the fiddler crab Uca lactea by providing males with additional food every other day. In food-supplemented enclosures, males did not increase courtship activity on the days when food was supplemented. However, they built more courtship structures (semidomes) and waved more on the days when they were not given additional food. Male size had a strong influence on the number of days the males courted. We also tested whether the frequency of surface mating, as an alternative reproductive tactic, decreased when food was supplemented. Contrary to our expectation, the number of males that exhibited the surface-mating tactic increased when food was supplemented whereas the number of mate-searching females did not change. Our findings in this field study suggest that reproductive decisions by male fiddler crabs are affected by fluctuating food availability and present body condition, and the alternative mating tactic of this species may be more frequently used by males under good condition.

Laboratory Culture of the Oriental Lancelet Branchiostoma belcheri

Abstract To overcome difficulties in getting research materials of cephalochordate lancelets, which has severely hampered experimental studies of this animal, we have attempted to establish a culture system in the laboratory. Adult animals collected from the wild were maintained in 2.5-L plastic containers filled with natural seawater without sand substratum. They were fed daily with unicellular algae. About 25% of the animals collected in 2003, 2004, and 2005 developed gonads in our culture system. Some of the sexually mature animals collected in the breeding seasons in 2005 and 2006 spawned spontaneously in the plastic containers of this system. Broods obtained in 2005 were maintained longer than a year in a glass tank without sand substratum. The progeny born in the laboratory showed great individual variation in growth but metamorphosed normally, and some of them started to develop gonads around 10 months after fertilization. Our mass culture methods for both adults and their progeny made daily observation possible and allowed the constant spawning of animals collected from the wild, at least in the summer season. Our culture method saves labor in maintenance and is easily set up without any specific demands except for running seawater, though still required to better survival rate and spawning control. Lancelet populations maintained in the laboratory can promote studies on these animals across disciplines and especially contribute to elucidation of the evolutionary history of chordates.

The effects of body size, ownership and sex-ratio on the precopulatory mate guarding of Caprella penantis (Crustacea: Amphipoda)

Precopulatory mate guarding behaviour of the skeleton shrimp Caprella penantis is described. Moreover, the effects of body size, ownership and sex-ratio on mate guarding were examined experimentally in the laboratory. In the field population, the operational sex-ratio was male-biased. Guarding pairs, which were collected from the field, continued guarding for an average of 350 minutes in the laboratory, indicating that the normal guarding duration is approximately 10 hours. In this species, two guarding types were found: Type O and Type I-like. In Type O guarding, the male would fold the female into a horseshoe shape, whilst the male held the female parallel to him in Type I-like guarding. In the laboratory experiments, male body size was the most important factor affecting competition for a receptive female; ownership was the secondary factor. Guarding duration was prolonged when the sex-ratio was male-biased. Thus, the precopulatory mate guarding behaviour of C. penantis is influenced by several factors, such as body size, ownership and sex-ratio.

Larval Development of the Oriental Lancelet, Branchiostoma belcheri, in Laboratory Mass Culture

Abstract We are successfully maintaining a laboratory colony of the lancelet Branchiostoma belcheri bred in the laboratory. Based on living individuals in this mass culture, morphological characteristics from the seven-day larval to benthic juvenile stages have been studied. Most striking was that later larval development of B. belcheri showed great individual variation even in a rather stable culture environment. Metamorphosis first occurred on 60 days post fertilization (dpf) and was continuously observed throughout the present study up to 100 dpf. Morphological traits such as the number of primary gill slits and body size at the start of metamorphosis are apparently affected by culture condition. Body size measured in the largest individuals showed nearly linear growth at 0.087 mm/day. The variability found in larval development calls for caution when developmental stages and chronological ages are compared between populations. However, the developmental flexibility of this animal also raises the possibility that growth and sexual maturation could be controlled artificially in captivity.

Stable aquaculture of the Japanese lancelet Branchiostoma japonicum for 7 years

Despite advances in the study on animal evolution in the last two decades, paucity of experimental data on cephalochordates comparable to those on the other chordates hinders an integrative understanding of chordate evolutionary history. To obtain lancelet data under well-controlled experiments, laboratory cultures of lancelets have been performed at several institutions. In a mass culture started in 2005, we have obtained up to three consecutive generations of Branchiostoma japonicum. Using sand substratum, survival rates of laboratory lancelets until maturation have improved to greater than 30%, much higher than compared to previously, and for adults the annual average survival rate was 82.3%. The high survival rate allows maintaining animals at least 6 years and potentially longer. Water temperatures lower than 23°C obviously reduced the frequency of spawning even after the onset of spawning period, and 1-2 days after changing the temperature at 25°C animals became spawned well. We also observed obvious sex reversal from male to female in individuals that had been cultured for 3 years or more. Our continuous culture has provided sufficient materials for vital experiments on early development and for studying metamorphosis, as well as for the conservation of wild populations. The subculture of successive laboratory generations will provide a valuable resource for genetic studies.

Evolutionary history of the extant amphioxus lineage with shallow-branching diversification

Amphioxus or lancelets have been regarded as a key animal in understanding the origin of vertebrates. However, the evolutionary history within this lineage remains unexplored. As the amphioxus lineage has likely been separated from other chordates for a very long time and displays a marked left-right asymmetry, its evolutionary history is potentially helpful in better understanding chordate and vertebrate origins. We studied the phylogenetic relationships within the extant amphioxus lineage based on mitochondrial genomes incorporating new Asymmetron and Epigonichthys populations, and based on previously reported nuclear transcriptomes. The resulting tree patterns are consistent, showing the Asymmetron clade diverging first, followed by the Epigonichthys and Branchiostoma clades splitting. Divergence time estimates based on nuclear transcriptomes with vertebrate calibrations support a shallow diversification of the extant amphioxus lineage in the Tertiary. These estimates fit well with the closure of seaways between oceans by continental drift, ocean currents, and present geographical distributions, and suggest a long cryptic history from the origin of amphioxus to its most recent diversification. Deduced character polarities based on phylogenetic analyses suggest that the common ancestor of the extant amphioxus existed in a tiny epibenthic state with larva-like appearance of extant amphioxus, likely with ciliate epidermis.

Description of Zoeae and Habitat of Elamenopsis ariakensis (Brachyura: Hymenosomatidae) Living within the Burrows of the Sea Cucumber Protankyra bidentata

Abstract The morphological characteristics of the zoeas and crabs of the false spider crab Elamenopsis ariakensis are described and their habitats are documented, with an emphasis on their symbiosis with the burrowing sea cucumber Protankyra bidentata. The 16 zoeas and 91 crabs were collected from the Ariake Sea and adjacent Yatsushiro Sea, Japan. Zoeas have no lateral spine, whereas the rostral and dorsal spines are extremely long. In all zoeal stages, the pleon consists of six segments, including the telson. Adult crabs have fused pleons, with five segments in males and three segments in females, including the telson, but six segments are found in young male crabs. All crabs were collected from P. bidentata burrows. In addition to E. ariakensis, seven commensal species were collected from P. bidentata burrows: two crabs, four bivalves, and one scale worm. The microhabitats in the burrow of P. bidentata were divided somewhat among species.

Fine scale meiofaunal distribution around burrows of ocypodoids (Decapoda, Ocypodoidea) in tidal flat sediments

In tidal flat sediments, larger invertebrates often modify the sedimentary environment in a manner that causes smaller animals, such as meiofauna, to become more abundant. In other words, macrofauna can “promote” meiofauna. In summer (August) and autumn (November/early December) 2007, we investigated centimetre-scale horizontal and vertical profiles of total metazoan meiofaunal abundance and community structure around burrows of Uca lactea (De Haan, 1835) (fiddler crabs) and Scopimera globosa (De Haan, 1835) (sand bubbler crabs) on an intertidal sandflat of the Amakusa Islands, western Japan. Meiofaunal community structure changed significantly with sediment depth around burrows of U. lactea in both seasons, whereas vertical differences were less clear for S. globosa. Although no evidence was found that total meiofaunal abundance or the abundances of certain taxonomic groups were positively influenced by burrows of S. globosa, the abundance of nematodes in subsurface sediment around burrows of U. lactea was significantly higher than in controls in autumn. High concentrations of fresh organic matter had not accumulated in the sediment around burrows of either species of crab. Uca lactea, however, made its burrows in significantly finer sediment than did S. globosa. One possibility is that differences in meiofaunal distribution in sediment around the burrows of the two crabs were the result of habitat and behavioural differences between the species.