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Impact of Different Diets on Adult Tri-Spine Horseshoe Crab,tachypleus tridentatus

Journal of Ocean University of China 2022年第3期21卷 541-548页

作者： YING Ziwei BAO Yuyuan LI Yinkang YE Guoling ZHANG Shuhuan XU Peng ZHU Junhua XIE Xiaoyong Guangdong Provincial Key Laboratory of Fishery Ecology and Environment South China Sea Fisheries Research InstituteChinese Academy of Fisheries SciencesGuangzhou510300China College of Fisheries Science and Life Science of Shanghai Ocean University Shanghai201306China Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou) Guangzhou511458China Guangdong Ocean Planning Research Center Guangzhou510322China Sturgeon Healthy Breeding and Medicinal Research Center Guizhou University of Traditional Chinese MedicineGuiyang550025China College of Marine Sciences Beibu Gulf UniversityQinzhou535011China

Effective culture and management of adult tri-spine horseshoe crab,tachypleus tridentatus can ensure that stock enhancement programs and aquaculture systems are maintained.To explore suitable feed for animals during the breeding season,Pacific oyster(Ostrea gigas)(oyster group;OG)and frozen sharpbelly fish(Hemiculter leucisculus)(frozen fish group;FG)were selected to feed 20 T.tridentatus male and female pairs,respectively.At the end of the experiment,intestinal samples were obtained to measure digestive enzymes activities.The intestinal flora were determined by 16S r DNA sequencing.No eggs were observed in the FG and one T.tridentatus adult died.No animals died in the OG,and 9.7×10;eggs were obtained.These results show that oysters are more suitable for the development and reproduction of adult T.tridentatus than frozen fish.Additionally,the digestive enzyme activity analysis revealed that animals in the OG exhibited higher protein digestibility than those in the FG,but no significant differences in lipid and carbohydrate uptake were observed between the groups.Furthermore,the intestinal flora analysis showed that operational taxonomic units(OTUs)and the Chao1 index were significantly higher in the OG than in the FG,but no significant difference was observed in the Shannon or Simpson indices between the groups.Our data indicate that the oyster diet improved the intestinal microbial diversity of T.tridentatus.We hypothesize that nutrients,such as oyster-based taurine,proteins,and highly unsaturated fatty acids,improve protease activity in the T.tridentatus digestive tract,alter the intestinal floral structure,and improve the reproductive performance of T.tridentatus.

关键词： tachypleus tridentatus diet reproductive performance digestive enzyme activity intestinal flora

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Effect of Intertidal Elevation at Tsuyazaki Cove,Fukuoka,Japan on Survival Rate of Horseshoe Crab tachypleus tridentatus Eggs

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Journal of Ocean University of China 2022年第3期21卷 601-610页

作者： ITAYA Shinji SHUUNO Mari ONIKURA Norio TAI Akira YANO Shinichiro Environmental Fluid Dynamics Laboratory Department of Urban and Environmental EngineeringGraduate School of EngineeringKyushu UniversityFukuoka819-0395Japan Tsuyazaki Seaside Nature School Fukutsu811-3304Japan Fishery Research Laboratory Department of Bioresource ScienceGraduate School of Bioenvironmental ScienceKyushu UniversityFukuoka819-0395Japan

Physical factors affecting the survival of tachypleus tridentatus eggs were investigated by translocating their eggs between the high intertidal zone and the low intertidal zone of a known spawning site.The mean egg survival rates per day were highest in the mid intertidal zone(45.1%±25.4%)and the lowest in the low intertidal zone(13.3%±27.6%).Differences in the elevation,air exposure time,and water content of the spawning ground were significant factors determining the egg survival rates.Excessive or insufficient air exposure time resulted in inadequate water content at higher and lower intertidal zones and could reduce egg survival.On the other hand,moderate saturation and dehydration were repeated with each tidal movement in the mid intertidal zone.This dynamic is considered as one of the crucial factors for the survival of eggs and is considered optimal for spawning.Therefore,the protection of the mid intertidal zone is imperative for maximizing the egg survival rate in Tsuyazaki Cove where almost all suitable nesting sites have disappeared due to coastal development.By protecting these optimal sites for spawning and recovering other optimal sites on suitable beaches,a positive contribution can be made to future management and conservation.The study also suggests that translocating eggs from marginal to optimal spawning sites might be a recovery strategy for this globally endangered species.

关键词： air exposure time globally endangered species intertidal elevation intertidal zone optimal spawning site tachypleus tridentatus translocated horseshoe crab eggs Tsuyazaki Cove

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Transcriptome Analysis of the Digestive Tract of tachypleus tridentatus and Carcinoscorpius rotundicauda

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Journal of Ocean University of China 2022年第3期21卷 591-600页

作者： LI Yuhong KWAN Kit Yue LI Sansu LIU Yingchun WENG Bosen HUANG Wenshu WENG Zhaohong Department of Environmental Science and Engineering Huaqiao UniversityXiamen361021China College of Marine Sciences Beibu Gulf UniversityQinzhou535011China School of Biomedical Sciences Huaqiao UniversityXiamen361021China School of Basic Medical Sciences Fujian Medical UniversityFuzhou350108China Fisheries College Jimei UniversityXiamen361021China

Transcriptome assemblies for the stomach,midgut,and fecal mucosa of tachypleus tridentatus and Carcinoscorpius rotundicauda are reported for the first time in this study.Genome assembly resulted in 373069 unigenes with an N50 of 1314,while transcriptome assembly resulted in 736378 unigenes and an N50 of 2121.The annotated unigenes showed the highest similarity to fishes,amphibians,and reptiles,with most unigenes closely related to metabolism,translation,biogenesis,signal transduction,energy production,immune response,and secretion.Unigenes(4768)were mapped to KEGG pathways,and the most enriched pathways were involved in translation,environmental information signal processing,metabolism,endocrine system,immune system,nervous system,and varying metabolism.Totally 65889,2001,and 7162 differentially expressed genes(DEGs)were identified in the stomach,midgut,and fecal mucosa between the two horseshoe crab species,respectively.Compared with C.rotundicauda,99.95%of the DEGs in the T.tridentatus stomach were down-regulated,while 99.97%of the DEGs of T.tridentatus fecal mucosa and 55.42%of the DEGs of the T.tridentatus midgut were up-regulated.Most midgut DEGs were involved in hydrolase activity,protein metabolism,and cell cytoplasm,while most stomach and fecal mucosa DEGs were involved in catalytic activity and primary metabolic processes.Most stomach DEGs were assigned to the cellular component in cellular macromolecular complexes,while most fecal mucosa DEGs were assigned to the cellular component in cell and cell parts.These results will benefit the investigation of the molecular mechanisms of the digestive tract related to the feeding habits and environmental traces of horseshoe crabs.

关键词： transcriptome digestive tract tachypleus tridentatus Carcinoscorpius rotundicauda differentially expressed genes(DEGs)

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Effect of Probiotics on Juvenile tachypleus tridentatus Gut Microbiota

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Journal of Ocean University of China 2022年第3期21卷 564-572页

作者： XIONG Kai MIAO Fengze WAIHO Khor LIU Ximei FANG James K.H. HE Jinfeng HU Menghong WANG Youji Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources Ministry of EducationShanghai Ocean UniversityShanghai201306China Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding Shanghai Ocean UniversityShanghai201306China Higher Institution Center of Excellence(HICoE) Institute of Tropical Aquaculture and FisheriesUniversity Malaysia TerengganuKuala TerengganuTerengganu20000Malaysia Department of Applied Biology and Chemical Technology The Hong Kong Polytechnic UniversityHung HomKowloon999077China Beihai Product Quality Testing Institute Beihai536000China

The population of tri-spine horseshoe crabs(HSCs)tachypleus tridentatus has sharply decreased in recent decades.Domestication and manual releasing are now used to recover it.In order to accelerate the growth of T.tridentatus in indoor environment,in this study,gut microbiota of 2nd instar T.tridentatus was supposed to be improved by putting probiotics in water environment and thus promoted the health and growth of the host.Using 16S rRNA sequencing of gut samples,we examined the intestinal microbiome of juvenile HSCs and studied the effect of probiotics.Results showed that there was no significant difference between groups fed with high and low concentrations(1×10^(7),1×10^(12)CFU mL^(−1))of Bacillus subtilis and high concentration(1×10^(12)CFU mL^(−1))of Lactobacillus,and no significant difference compared with the control group,but the diversity of Lactobacillus group with low concentration(1×10^(7)CFU mL^(−1))was significantly higher than those of other groups.Microbial-mediated functions predicted by PICRUSt showed that‘Cellular Processes’,‘Metabolism’and‘Organismal Systems’pathways significantly increased in groups of Bacillus subtilis and Lactobacillus group with low concentration.The results showed that two types of probiotics with different concentrations have other effects on the structure,richness and function of intestinal microbes.By searching the literature and conducting pre-experiments,we forecasted the short time of breeding under the probiotic environment may not have much effect.The present study for the first time investigated the impact of probiotics on gut microbiota of juvenile HSCs and found them may benefit for the growth and health of HSCs.

关键词： tachypleus tridentatus probiotic gut microbiota diversity richness

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Indiscriminate Dietary Compositions of Two Asian Horseshoe Crabs,tachypleus tridentatus and Carcinoscorpius rotundicauda:Evidence from Hemolymph Stable Isotopes

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Journal of Ocean University of China 2022年第3期21卷 583-590页

作者： LI Yuhong FENG Jianxiang KWAN Kit Yue TAN Yiran WENG Bosen JIA Ruiling Department of Environmental Science and Engineering Huaqiao UniversityXiamen361021China School of Marine Sciences Sun Yat-sen UniversityZhuhai519082China College of Marine Sciences Beibu Gulf UniversityQinzhou535011China

Knowledge of dietary compositions is essential to the recovery of the dwindling populations of horseshoe crabs in China.The feeding habits of tachypleus tridentatus and Carcinoscorpis rotundicauda of Pearl Bay in southwestern China were studied by stable isotope analysis.Hemolymph samples of the two species were obtained from two age groups living in different habitats(inner,central,and outer Bay).In addition,their potential food source samples were collected in May 2019.Results showed that the mean values ofδ^(13)C andδ^(15)N in C.rotundicauda individuals ranged from−19.01‰to−16.47‰and from 10.49‰to 13.5‰,respectively,and those of T.tridentatus ranged from−19.12‰to−14.96‰and from 8.78‰to 13.48‰,respectively.These values indicated that the horseshoe crabs have a wide variety of food sources and therefore are highly omnivorous.No remarkably correlations were found betweenδ^(13)C andδ^(15)N values and individual widths in the two species.Thirteen potential food sources were selected,in which Sipunculus nudus,Batillaria zonalisi,and Ulva lactuca contributed largely to the dietary compositions of the two species of horseshoe crabs.No significant correlation was found between the feeding habits and habitat geochemical characteristics of the horseshoe crabs.All these results possibly indicate the high diversity of their food sources and the indiscriminate dietary compositions of the two Asian horseshoe crabs.

关键词： tachypleus tridentatus Carcinoscorpius rotundicauda stable isotope hemolymph growth stage

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Ontogenetic resource use and trophic dynamics of endangered juvenile tachypleus tridentatus among diversified nursery habitats in the northern Beibu Gulf,China

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Integrative Zoology 2021年第6期16卷 908-928页

作者： Kit Yue KWAN Justin BOPP Shuyan HUANG Qiao CHEN Chun-Chieh WANG Xueping WANG Wenquan ZHEN Junhua ZHU Xing HUANG College of Marine Sciences Beibu Gulf UniversityQinzhouChina Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation QinzhouChina School of Marine and Atmospheric Sciences Stony Brook UniversityStony BrookNew YorkUSA Futian-CityU Mangrove R&D Center City University of Hong Kong Shenzhen Research InstituteShenzhenChina Guangxi Key Laboratory of Marine Environmental Science Guangxi Beibu Gulf Marine Research CenterGuangxi Academy of SciencesNanningChina Guangxi Key Laboratory of Marine Disaster in the Beibu Gulf QinzhouChina

Horseshoe crabs,the most well-known example of“living fossils”,are iconic and ecologically important macroinvertebrates in coastal and estuarine ecosystems.Their blood is a crucial resource for manufacturing Limulus or tachypleus amebocyte lysate to detect bacterial endotoxins or fungal contamination in drug and medical devices.An enhanced understanding of their ecological roles and trophic interactions in the food webs is fundamental to facilitate resource management for the declining populations in Asia.Foraging information of the Asian species,however,is mainly derived from preliminary,scattered reports from a limited number of study locations.In this study,resource utilization,trophic niche dynamics,and trophic interaction of the juvenile tri-spine horseshoe crab,tachypleus tridentatus(instars 1–12,approximately 0.5–8 years old)across ontogeny was assessed in diversified nursery habitats along the northern Beibu Gulf,China,using carbon and nitrogen stable isotopes.Our results suggest that:(i)T.tridentatus are ecological generalists given the vast range of carbon isotopic values and trophic niche width estimates exhibited between multiple instar groups;(ii)juvenile T.tridentatus across most habitat types predominantly assimilated energy from a variety of basal production sources in the food web,but primarily depended on sedimentary organic matter and seagrass resource pools;(iii)ontogenetic shifts in juvenile dietary proportions were evident,with decreased reliance on sedimentary organic matter,coupled with increased reliance on benthic macroinvertebrate grazers,detritivores,and omnivores with age;and(iv)nearly all juvenile instars occupied similar trophic positions in the food web with slight shifts in trophic position present with increasing size.Our findings indicate that resource availability and ontogenetic diet shifts strongly influence horseshoe crab trophic dynamics,and age should be accounted when formulating habitat conservation measures based on resource use for Asian horseshoe crabs.

关键词： intertidal food webs ontogenetic shift stable isotopes tachypleus tridentatus trophic niche width

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Spatiotemporal Distribution of Asian Horseshoe Crab Eggs Are Highly Intermingled with Anthropogenic Structures in Northern Beibu Gulf,China

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Journal of Ocean University of China 2022年第3期21卷 531-540页

作者： KWAN Kit Yue FU Yijian ZHONG Mufeng KUANG Yang BAI Haiwei ZHANG Ce ZHEN Wenquan XU Peng WANG Chun-Chieh ZHU Junhua College of Marine Sciences Beibu Gulf Ocean Development Research CenterBeibu Gulf UniversityQinzhou535011China Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation Qinzhou535011China Guangxi Key Laboratory of Marine Disaster in the Beibu Gulf Qinzhou535011China Guangxi Beibu Gulf Marine Research Center Guangxi Academy of SciencesNanning530007China

Identification,protection and restoration of spawning habitats are vital for protecting the depleted species.Asian horseshoe crabs are ecologically important macroinvertebrates in coastal and estuarine ecosystems.However,their spawning habitat studies were limited to several reports in tropical regions,possibly due to the lack of modified survey methods,particularly in habitats with a lower density of spawning adults,and/or intermingled with anthropogenic structures.In this study,the year-round egg distribution and spawning habitat baselines of Carcinoscorpius rotundicauda and tachypleus tridentatus were determined in the northern Beibu Gulf,China.Our findings demonstrated that the peak spawning occurred in June–July and ceased in November–January when the average water temperature dropped below 20℃.Egg aggregations were found<10 cm beneath the sediment surface with regular tidal inundation,regardless of seasonal changes,in the vicinity of natural and artificial structures with elevated,mildly sloping substratum within the high tide zones.The nests were characterized by medium-sized sediment grains(0.5–0.9 mm),high temperatures(31–34℃),low water contents(0.8%–0.9%),and total organic carbon contents(0.5%–0.7%),which might maximize the hatching success.The identified nesting beaches were close to nursery habitats for juveniles,and tidal creeks were present as the possible corridor connecting these two important habitats through the dominant mangrove forests.The findings provide valuable insights in the scope of spawning behavior and nest-site selection of Asian horseshoe crabs under a mixture of natural and artificial structures,which could benefit future management efforts for the exploited spawning populations.

关键词： Carcinoscorpius rotundicauda tachypleus tridentatus mangrove tidal creek sediment texture slope elevated substratum

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Isolation and characterization of microsatellite loci in the horseshoe crab(tachypleus tridentatus)

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广西水产科技 2014年第1期 8-8页

作者： Qiong-Zhen Li Qi Li Jun Liu Le-Hai Ni Ling-Feng Kong

tachypleus tridentatus isan endangered horseshoe crab that isfound and known only in several isolated sites in southern China,and thathas been suffering from the loss and fragm entation of its habitat.To evaluate its genetic diversity and population genetics,the first set of eight polym orphicm icrosatellite lociwas developed and characterized.The number of alleles ranged from 3 to18 per locus with the observed and expected heterozygosities ranging from 0.667 to 1.000 and from 0.543 to 0.944,respectively.These markers will be useful for the assessment of genetic variation for this species.

关键词： tachypleus tridentatus Horseshoe crab Microsate llites

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Activity Recording of Free-Ranging Tri-spine Horseshoe Crabs in the Southeastern Coast of Sabah,Borneo

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Journal of Ocean University of China 2022年第3期21卷 549-556页

作者： WATANABE Shinichi NELSON Bryan Raveen MANCA Azwarfarid MOHAMAD Faridah Little Leonardo Co. Tokyo1130021Japan Institute of Tropical Biodiversity and Sustainable Development Universiti Malaysia TerengganuKuala NerusKuala Terengganu21030Malaysia Pertubuhan Pelindung Alam Malaysia Casa Green CherasAlam43200Malaysia Horseshoe Crab Research Group Faculty of Science and Marine EnvironmentUniversiti Malaysia TerengganuKuala NerusKuala Terengganu21030Malaysia

In Inderasabah(southeast Sabah),tri-spine horseshoe crab tachypleus tridentatus was observed for their locomotion activity using data loggers from September to November in 2015.A female with acceleration and depth-temperature loggers and five males with acceleration loggers were recaptured between 10 and 49 days after their release.From the record of 194 activity days that involve all six T.tridentatus,four horseshoe crabs,including the female,were active throughout the 24.0 h cycle,whereas the activity of the remaining two males was consistent with the 12.4 h cycle.Using the 40-day recording,three horseshoe crabs,including the female,were primarily active around the new moon and full moon,but they were dormant around the first and third quarter moon days.The female spent much time in shallow shores(depthtridentatus in Inderasabah was active during daytime and nighttime.This result was contrary to T.tridentatus activity cycles in western Japan,where the species was found to be primarily nocturnal.Perhaps,the regional differences in activity cycles for T.tridentatus were related to their population adaptation toward water temperature,depth,and prey-searching periods.

关键词： acceleration logger activity cycle depth-temperature logger foraging behavior spawning behavior tropical region tachypleus tridentatus

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Asian Horseshoe Crab Bycatch in Intertidal Zones of the Northern Beibu Gulf:Suggestions for Conservation Management

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Journal of Ocean University of China 2022年第3期21卷 611-621页

作者： WANG Chun-Chieh CHEN Ruifang YANG Xin WEN Yulong KUANG Yang ZHANG Ce ZHU Junhua KWAN Kit Yue Guangxi Key Laboratory of Marine Environmental Science Guangxi Beibu Gulf Marine Research CenterGuangxi Academy of SciencesNanning530007China Guangxi Key Laboratory of Beibu Gulf Marine Biodiversity Conservation College of Marine SciencesBeibu Gulf UniversityQinzhou535011China

Bycatch poses a severe challenge to global fishery management.Although studies have focused on marine megafauna bycatch,research on relevant issues regarding invertebrates is limited,particularly for the threatened horseshoe crabs.In this study,the distribution of fishing gears and the bycatch intensity of Asian horseshoe crabs in the intertidal zones of the northern Beibu Gulf that harbors the most abundant juvenile Asian horseshoe crabs populations in China were evaluated.Seven intertidal nursery habitats for tachypleus tridentatus and Carcinoscorpius rotundicauda were surveyed from May to July in 2021.A transect that crossed the tidal creeks and tidal flats between the high tide embankment/vegetation and low tide line of a given habitat was surveyed during the ebb tides.The type,number,and GPS of fishing gears were recorded when sighted,and the number and prosomal width of each horseshoe crab species were measured.Bycatch intensities differed among habitats,ranging from 0.3 to 18.4 and 1.2 to 22.7 individuals per kilometer of transect for T.tridentatus and C.rotundicauda,respectively.Among the three identified fishing gear types,ground cages and stick net sets caused a stronger bycatch pressure on these two species.Most T.tridentatus bycatch and C.rotundicauda bycatch were large individuals in late juvenile and adult stages.Therefore,the ground cages and stick net sets along/near the tidal creeks,mangrove fringe,and low tide line should be regulated and removed to ensure the functionality of the intertidal zone as the spawning corridor and nursery habitat of Asian horseshoe crabs.

关键词： tachypleus tridentatus Carcinoscorpius rotundicauda ground cage stick net tidal creek mangrove low tide line spawning corridor

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