Assessment of the Characteristics of Demersal Fish Communities Using Species-and Trait-Based Approaches in the Coastal Habitats Along Rongcheng Bay,Shandong Peninsula,China

Biodiversity declines have motivated many studies on the relationship between species diversity and ecosystem functioning.In this study,we described the spatial-temporal characteristics of demersal fish communities along a coastal habitat in Rongcheng Bay,Shandong Peninsula,China with both species-based and biological trait-based approaches.The field survey was carried out monthly using traps from April to October of 2018,and divided into three seasons(spring:April and May;summer:June,July and August;autumn:September,October and November).The study area included five distinct habitats:seagrass bed,natural rocky reef,bare sand,artificial reef together with natural rocky reef,and artificial reef together with bare sand.We analyzed the fish communities with three taxonomic diversity indices,including Shannon-Wiener,Simpson,and Pielou Evenness,as well as four functional diversity indices,including FRic,FEve,FDiv,and FDis,based on 7 functional groups which are categorized into 27 traits.The results showed that there were no significant differences in taxonomic diversity indices among different habitats in the three seasons.However,significant differences were found in the functional richness of fish communities among different habitats in three seasons.Seagrass bed represented the highest functional richness in spring and autumn.This study demonstrates that seagrass bed is very important in enhancing the functional diversity of fish communities in a complex habitat.The study also indicates that the combination of taxonomic diversity and functional diversity will provide a more detailed description of the characteristics of fish communities.

Impacts of Integrated Multi-Trophic Aquaculture on Phytoplankton in Sanggou Bay

Integrated multi-trophic aquaculture(IMTA)has been considered as an ecofriendly culture system providing a potential solution to environmental risks caused by intensive monoculture system.However,the impact of IMTA on phytoplankton remains unclear.In this study,the spatial and temporal variations of phytoplankton in Sanggou Bay were investigated seasonally based on 21 sampling sites covering three cultivation zones(bivalve zone,IMTA zone,and kelp zone)and one control zone(without aquatic cultivation).In total,128 phytoplankton species,with diatoms and dinoflagellates as the dominant groups,were obtained across the whole year,and the mean Shannon diversity index(H')and species richness(SR)were determined as 1.39 and 9.39,respectively.The maximum chlorophyll a(Chl-a)(6.32μg L^(-1))and plankton diversity(H'of 1.97)occurred in summer and autumn,respectively.Compared to other zones,the bivalve zone displayed significantly higher Chl-a and lower H'in majority of time.Pairwise PERMANOVA analysis indicated that the phytoplankton assemblage in the bivalve zone was significantly different with the control and kelp zones,while the IMTA zone maintained close to other three zones.Based on generalized additive models,temperature,NO_(2)^(-)-N,N/P ratio,SiO_(3)^(2-)-Si,and salinity were determined as the key factors underlying Chl-a and phytoplankton diversity.Addi-tionally,the results of redundancy analysis further indicated that the phytoplankton assemblage in the bivalve zone is positively re-lated with nutrients such as NO_(3)^(-)-N and NH_(4)^(+)-N as well as water depth,while the phytoplankton assemblages in the kelp,control,and IMTA zones are associated with NO_(2)^(-)-N,SiO_(3)^(2-)-Si,and salinity.Taken all observations into consideration together,it can be inferred that IMTA can effectively reduce Chl-a level compared to bivalve monoculture by reducing the nutrients.However,the SR,H’,and species composition of phytoplankton are primarily determined by local environment factors such as temperature,water depth,salinity and SiO_(3)^(2-)-Si.

Discordant patterns of genetic variation between mitochondrial and microsatellite markers in Acanthogobius ommaturus across the coastal areas of China

Acanthogobius ommaturus, which belongs to the family Gobiidae, is a euryhaline and demersal fish that is widely distributed in the coastal areas, harbors, and estuaries of China, D. P. R. Korea and Japan. In this study, the genetic diversity and genetic structure of five geographical populations of A. ommaturus was assessed using the mitochondrial hypervariable region gene and microsatellite markers. The results of the two genetic markers indicated that the A. ommaturus populations had a high level of genetic diversity. The mitochondrial marker detected weak genetic differentiation among populations, and the Neighbor-Joining tree showed that there was no obvious pedigree branches and geographic structure as well. However, population of Zhoushan showed significant genetic differentiation with other populations by microsatellite markers. The population of A.ommaturus has not experienced bottleneck effect recently. We speculated that the Pleistocene climate change and juvenile fish dispersal played an important role in the population differentiation of A. ommaturus.

Characterization of Caspase Gene Family Members in Spotted Sea Bass(Lateolabrax maculatus)and Their Expression Profiles in Response to Vibrio harveyi Infection

The caspase gene family is a crucial gene cluster that regulates apoptosis which contribute to programmed cell death,cell proliferation and differentiation,and several immune responses.In our study,a complete set of 12 caspase genes were identified in spotted sea bass Lateolabrax maculatus.These genes were divided into three subfamilies:2 inflammatory caspases(casp-1 and casp-14-like),5 apoptosis initiators(casp-2,casp-8a,casp-8b,casp-9,and casp-10),and 5 apoptosis executioners(casp-3a,casp-3b,casp-3-like,casp-6,and casp-7).Their phylogenetic relationships,synteny and gene structures were systematically analyzed.Furthermore,the relative expression profiles of the caspase family members in the liver,intestine,head kidney,and spleen were measured by q PCR after infection with Vibrio harveyi.The results showed that the overall mRNA levels of the caspase genes were dramatically increased after V.harveyi infection,and the expression patterns varied among genes and tissues.More caspase genes underwent pronounced expression changes in the head kidney and spleen than in the liver or intestine,mainly after 48 h of the challenge.Specifically,casp-3a,casp-3b,casp-3-like,casp-6,casp-7,casp-8a,casp-8b,casp-10,and casp-14-like in the head kidney,and casp-3-like,casp-6,casp-7,and casp-14-like in the spleen,were the most responsive caspase genes which may contribute significantly to immune regulation in spotted sea bass.Additionally,the apoptosis level in head kidney and spleen after infection were examined using the Caspase assay.Our study provides a systemic overview of the caspase gene family in spotted sea bass after V.harveyi infection and lays a foundation for further deciphering the biological roles of these caspase genes.

Genome-wide characterization of mapk gene family in black rockfish Sebastes schlegelii and their expression patterns against Edwardsiella piscicida infection

Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in teleost.Identification,characterization,classification,and expression profiling of totally 15 mapk genes in black rockfish(Sebastes schlegelii)were conducted.Phylogenetic relationships show that these mapk genes could be divided into extracellular signal-regulated kinase(ERK),c-Jun N-terminal kinase(JNK),and p38 sub-families.In addition,gene structures,syntenic analysis,and selective pressure analysis are performed to confirm their annotations.Results of selective pressure analysis indicate that mapk1,mapk3,mapk7,mapk10,mapk11,and mapk12 underwent significantly-positive selections,while the others genes such as mapk4,mapk6,mapk15,mapk8a,mapk8b,mapk9,mapk13,mapk14a,and mapk14b were under purifying selections.Moreover,results of qRT-PCR indicate that mapk genes in 8 healthy tissues displayed different expression patterns.The expression patterns of several mapk genes including mapk12,mapk13,mapk14a,mapk14b,and mapk15 were significantly changed in mucosal tissues after Edwardsiella piscicida infection.This study demonstrates that mapk genes in black rockfish play vital prevention roles against bacterial infection,which not only helps us understand the structure and function of mapk genes in black rockfish,but also provides a reference to understand the role of mapk genes in teleost immune responses.

Ultrastructural variation and key ER chaperones response induced by heat stress in intestinal cells of sea cucumber Apostichopus japonicus

Abstract The unfolded protein response(UPR)is an important protective and compensatory strategy used during endoplasmic reticulum stress caused by factors including glucose starvation,low pH,or heat shock.However,there is very little information on the possible role(s)of the UPR under adverse conditions experienced by marine invertebrates.We observed that rough endoplasmic reticulum(ER)was dramatically expanded and numerous autophagosomes were accumulated in the intestinal cells of sea cucumbers,Apostichopus japonicus,under heat stress(4 h at 25°C compared with 15°C controls).Moreover,heat stress led to sharp increases in the relative transcript and protein expression levels of two primary ER chaperones:the endoplasmic reticulum resident protein 29-like(ERP29)and protein disulfi de-isomerase A6-like(PDIA6).These results suggest a potential adaptive mechanism to deal with heat-induced stress in sea cucumber intestine.

Prey preference of the common long-armed octopus Octopus minor(Cephalopoda: Octopodidae) on three different species of bivalves

Octopus minor is widely distributed along the northern coast of China. To date, there is little information on the prey selection process of this species. To understand this process, several experiments were carried out. Three types of bivalves, namely, Ruditapes philippinarum , Mactra chinensis , and Mytilus galloprovincialis , were used to observe the prey selection of O . minor and to analyze the potential causes of prey selection from three aspects: prey profi tability, adductor muscle tension and handling time. Under single-prey conditions, we found that the average (±SD) predation rates of O . minor on R . philippinarum , M . chinensis , and M . galloprovincialis were 1.73±0.50, 1.27±0.42, and 0.8±0.2/d, respectively. Under diff erent prey combinations, octopods actively selected one type of prey over the other(s), and the order of prey preference was R . philippinarum , followed by M . chinensis and lastly M. galloprovincialis . Furthermore, the shells of the consumed prey showed that O . minor only consumed bivalves by pulling them apart since there was no evidence of drill holes on the shells. The prey selection of O . minor was related to the prey profi tability and handling time;O . minor appeared to select preys with a higher profi tability and a shorter handling time. However, the diffi culty in opening the bivalve was not consistent with the prey preference of the octopods. These results suggest that O . minor prefers to consume R . philippinarum possibly due to a high profi tability and a short handling time that supports the optimum Foraging Theory.

Stocking density affects the growth performance and metabolism of Amur sturgeon by regulating expression of genes in the GH/IGF axis

The effects of stocking density on the growth and metabolism of Amur sturgeon were assessed. Amur sturgeon were grown for 70 days at three dif ferent stocking densities(low stocking density, LSD: 5.5 kg/m^3; medium stocking density, MSD: 8.0 kg/m^3; and high stocking density, HSD: 11.0 kg/m^3), and the biometric index, muscle composition, and serum biochemical parameters were evaluated. In addition, pituitary, liver, and muscle samples were collected for gene cloning and expression analyses. After 70 days of growth, the fish maintained at HSD had significantly lower fi nal body weight and specifi c growth rate, and a higher feed conversion ratio than those of the fish in the MSD and LSD groups. The HSD group had the lowest lipid and protein concentrations in serum and muscle. The serum cortisol concentration increased significantly in the HSD group, indicating that the stress-response system was activated in these fish. There was no change in the concentration of serum insulin-like growth factor 2(IGF-2), while the concentrations of serum growth hormone(GH) and insulin-like growth factor 1(IGF-1) decreased in the HSD group. The full-length cDNAs of G H and IGF-2 genes(995-bp and 1 207-bp long, respectively), were cloned and analyzed. In the HSD group, the expressions of GH in the pituitary and growth hormone receptor( GHR) and IGF-1 in the liver were down-regulated at the end of the 70-day experiment. In the HSD group, the transcript level of IGF-2 significantly decreased in the liver, but did not change in muscle. Overall, our results indicated that a HSD negatively af fects the growth performance and leads to changes in lipid and protein metabolism in Amur sturgeon. The down-regulated expression of genes related to the GH/IGF axis may be responsible for the poor growth performance of Amur sturgeon under crowding stress.

Isolation of CYP17 Ⅰ, 3β-HSD and AR Genes from Spotted Sea Bass (Lateolabrax maculatus) Testis and Their Responses to Hormones and Salinity Stimulations

In this study, three cDNA sequences corresponding to cytochrome P450 C17(CYP17 Ⅰ), 3β-hydroxysteroid dehydrogenase(3β-HSD) and androgen receptor(AR) were isolated from spotted sea bass(Lateolabrax maculatus). The mRNA abundances of CYP17 Ⅰ and 3β-HSD increased from stage Ⅱ to stage V with a significant increase at stage Ⅴ, and the highest abundance of AR mRNA was detected at stage Ⅲ in testicular development cycle. CYP17 I, 3β-HSD and AR transcripts were obviously abundant in steroidogenesis tissues such as testis, brain, head kidney among others. Strong and positive signals were observed mainly in interstitial cell regions of L. maculatus testis as were measured with in situ hybridization method. Significant increases of CYP17 Ⅰ and3β-HSD transcripts were detected after 12-48 h hCG(human chorionic gonadotropin) and GnRHa(gonadotropin-releasing hormone analogue) treatments. However, an opposite relationship was found for AR in testis at the same time. In addition, decreasing trends of CYP17 Ⅰ and 3β-HSD mRNA were observed in testis of L. maculatus in freshwater group(FW) from day 2 to day 6, and mRNA abundance of AR increased in brackish water(BW) group from day 4 to day 8. These findings revealed that these three steroid synthesis genes are import for testicular development, hormone and salinity treatment, and provided also an insight into the mechanism of reproductive endocrine of L. maculatus.

Fatty Acid Composition and Digestive Enzyme Activities of Rainbow Trout in Response to Dietary Docosahexaenoic Acid(DHA)and Eicosapentaenoic Acid(EPA)During Salinity Acclimation

This physiological study aimed to evaluate the effects of dietary docosahexaenoic acid(DHA)and eicosapentaenoic acid(EPA)on the fatty acid composition and digestive enzyme activities of rainbow trout(Oncorhynchus mykiss)during salinity acclimation.Rainbow trout with an average initial weight of 90.61 g±9.25 g were fed diets with the quantities of DHA and EPA equaling to 0.54%,0.95%,1.40%and 1.79%(abbreviated as DE-0.54,DE-0.95,DE-1.40,and DE-1.79,respectively)for eight weeks,after which the gastric and intestinal fatty acids composition were analyzed.Subsequently,the fish underwent salinity acclimation.On days 1,4,7,and 14 after the freshwater was replaced by seawater and at the end of the 8-week period,gastric and intestinal digestive enzyme activities were determined.The results showed that the gastric and intestinal DHA and EPA contents of the fish were positively correlated to their dietary DHA and EPA levels.Low dietary DHA and EPA levels inhibited the protease activity of rainbow trout.Fish in the DE-0.54 group increased the lipase activity to enhance the utilization of lipids maybe due to the inadequate essential fatty acids for fish in this group.Hence,rainbow trout in the DE-0.54 group failed to maintain suitable activities of digestive enzymes after salinity acclimation.Therefore,a diet with minimum 0.95%DHA and EPA levels is necessary for rainbow trout during salinity acclimation.

Mating behaviors in ovoviviparous black rockfish(Sebastes schlegelii):molecular function of prostaglandin E2 as both a hormone and pheromone

Prostaglandins(PGs)are profound hormones in teleost sexual behavior,especially in mating.PGs act as pheromones that affect the olfactory sensory neurons of males,inducing the initiation of a series of mating behaviors.However,the molecular mechanism by which PGs trigger mating behavior in ovoviviparous teleosts is still unclear.In the present study,we employed the ovoviviparous black rockfish(Sebastes schlegelii),an economically important marine species whose reproductive production is limited by incomplete fertilization,as a model species.The results showed that when the dose of PGE2 was higher than 10 nmol/L,a significant(P<0.05)increase in mating behaviors was observed.Dual-fluorescence in situ hybridization indicated that PGE2 could fire specific neurons in different brain regions and receptor cells in the olfactory sac.After combining with specific neurons in the central nervous system(CNS),a series of genes related to reproduction are activated.The intracerebroventricular administration of PGE_(2) significantly increased lhb levels(P<0.05)in both sexes.Moreover,steroidogenesis in gonads was also affected,inducing an increase(P<0.05)in E_(2) levels in males and T levels in females.PGE_(2) levels were also increased significantly(P<0.05)in both sexes.The present study revealed that PGE2 can activate mating behavior in black rockfish in both hormone and pheromone pathways,leading to variations in sex steroid levels and activation of reproductive behaviors.Our results provide not only novel insight into the onset of mating behaviors in ovoviviparous teleosts but also solutions for the incomplete fertilization caused by natural mating in cage aquaculture.

A paradox： rapid evolution rates of germline-limited sequences are associated with conserved patterns of rearrangements in cryptic species of Chilodonella uncinata(Protista, Ciliophora)

Some of the most extreme genome wide rearrangements are found in ciliates, which are unique in possessing both germline micronucleus and somatic macronucleus in every cell/organism. A series of DNA rearrangement events, including DNA elimination, chromosomal fragmentation, gene unscrambling and alternative processing, happen during macronuclear development. To assess the molecular evolution of macronuclear and germline-limited sequences in different cryptic species of Chilodonella uncinata, we characterized the actin, α-tubulin and β-tubulin genes in the micronucleus and macronucleus genomes of USA-SC2 strain and compared them with other strains(i.e. cryptic species). Three main results are:(i) rearrangement patterns between germline and soma are conserved for each gene among the cryptic species of C. uncinata;(ii) in contrast, the germlinelimited regions are highly divergent in sequence and length among the cryptic species;(iii) pointer shifting is frequent among the cryptic species. We speculate that pointer sequences may serve as the buffer between the conserved macronuclear destined sequences and rapidly-evolving internal eliminated sequences. The data combined with previous studies demonstrate the plasticity of gene rearrangement among different groups of ciliates and add to the growing data for the role of genome rearrangements in species differentiation.

Neuropeptide Y and melanocortin receptors in fish:regulators of energy homeostasis

Energy homeostasis,which refers to the physiological processes that the energy intake is exquisitely coordinated with energy expenditure,is critical for survival.Therefore,multiple and complex mechanisms have been involved in the regulation of energy homeostasis.The central melanocortin system plays an important role in modulating energy homeostasis.This system includes the orexigenic neurons,expressing neuropeptide Y/Agouti-related protein(NPY/AgRP),and the anorexigenic neurons expressing proopiomelanocortin(POMC).The downstream receptors of NPY,AgRP and post-translational products of POMC are G protein-coupled receptors(GPCRs).This review summarizes the compelling evidence demonstrating that NPY and melanocortin receptors are involved in energy homeostasis.Subsequently,the comparative studies on physiology and pharmacology of NPY and melanocortin receptors in humans,rodents and teleosts are summarized.Also,we provide a strategy demonstrating the potential application of the new ligands and/or specific variants of melanocortin system in aquaculture.