Significance of different microalgal species for growth of moon jellyfish ephyrae, <Emphasis Type="Italic">Aurelia</Emphasis> sp.1

The scyphozoan Aurelia aurita (Linnaeus) sp. l., is a cosmopolitan species-complex which blooms seasonally in a variety of coastal and shelf sea environments around the world. The effects of different microalgal species on the growth of newly-released Aurelia sp.1 ephyrae were studied under laboratory conditions. We fed ephyrae with four different microalgal species (diatom, autotrophic dinoflagellate, heterotrophic dinoflagellate, and chlorophyta) plus Artemia nauplii for 12-24 d at 18°C. Results showed that the growth rate diverged significantly for Artemia nauplii compared to other food types. In addition, there was no significant variation between the growth rates for Skeletonema costatum and Prorocentrum donghaiense, and no significant variation was found in the growth rates for N. scintillans and P. subcordiformis. Artemia nauplii could support the energy requirement for the newly-released ephyrae to develop to meduase, and the ephyrae with Artemia nauplii showed a significant average growth rate of 25.85% d^?1. Newly-released ephyrae could grow slightly with some species of microalgae in the earliest development stage. Chain diatom Skeletonema costatum and autotrophic dinoflagellate Prorocentrum donghaiense, could not support the growth of the ephyrae, while heterotrophic dinoflagellate Noctiluca scintillans and chlorophyta Platymonas subcordiformis could support the growth of the ephyrae. However, none of the ephyrae fed with the tested phytoplankton could mature to medusae.     

Biodegradation of <Emphasis Type="Italic">Enteromorpha</Emphasis> polysaccharides by intestinal micro-community from <Emphasis Type="Italic">Siganus oramin</Emphasis>

Micro-communities are supposed to have more potential functions of biodegradation of polysaccharides than single strain; however, the intestinal micro-communities involved in the biodegradation of Enteromorpha polysaccharides (EP) were seldom reported. In order to obtain the EP-degrading micro-community, the intestines of Siganus oramin was obtained to isolate the micro-communities, which were enriched by 0.3% of EP as the sole carbon source. A stable micro-community with EP degradative capability was achieved after seven generations of subculture, named H1. Results showed that H1 was able to degrade 75% of EP within 24 hours, and the activity of EP lyases reached 500 U mL^?1 in 32 hours. With denaturing gradient gel electrophoresis (DGGE) and 16S rRNA gene clone library analysis, ten bacteria closely related to Marinomonas pontica, Microbacterium sp., Leucobacter chironomi, Cyclobacterium sp., Algoriphagus winogradskyi, Pseudoalteromonas sp. and Vibrio sp. were determined. Furthermore, compared with the DGGE bands sequence and the clone library analysis, the dominant bacteria of the EP-biodegrading micro- community were Pseudoalteromonas sp. and Vibrio sp., with the respective proportion of 38% and 46%, and they should play an important role in EP degradation together with other degrading bacteria in the micro-community H1.     

Genotype and Phylogenetic Diversity of <Emphasis Type="Italic">Symbiodinium</Emphasis> ITS2 Sequences Within Clade C in Three Typical Coral Species from Luhuitou Fringing Reef of the South China Sea

Dinoflagellates in the genus Symbiodinium, including nine clades (A–I), mainly form mutualistic symbioses with corals. More than 100 Symbiodinium molecular types have been identified by the ITS2-based genotype method within any given clade, and specifically within Symbiodinium clade C. However, the genotype identification method using the ITS2 sequence is likely to lead to high diversity estimates due to the intra-genomic variations in the ITS2 space; thus, further validation is essential for a correct identification. In this study, the molecular diversity of Symbiodinium ITS2 sequences cloned from two stone corals, Acropora sp. SY-01 and Pocillopora sp. SY-05, and one soft coral, Sarcophyton sp. SY-07, living in the northern part of South China Sea (SCS), were analyzed and compared using the ITS2-based genotype identification method, coupled with ITS2-based secondary structural and phylogenetic analyses. As the result, 12 Symbiodinium ITS2 genotypes were identified, while only six and three Symbiodinium ITS2 genotypes were supported by ITS2-based secondary structural and phylogenetic analyses, respectively. In addition, no shared Symbiodinium ITS2 genotypes were observed among the three coral species, suggesting coral species-dependent Symbiodinium genotypes were within clade C. In summary, the present study provides a theoretical basis for validating the molecular diversity of Symbiodinium ITS2 genotypes in corals.     

Antifouling effect of bioactive compounds from selected marine organisms in the Obhur Creek,Red Sea

Three species of sponges and a tunicate were collected from Obhur creek of Jeddah coast for this bioactivity study. In order to assess the antifouling efficacy of selected marine organisms, methanolic extracts of these organisms were tested against different fouling bacterial forms and II-instar stage of the barnacle, Balanus amphitrite. Antibiosis, bioactivity and followed by multivariate analyses were carried out to check the efficacy of antifouling effect of the selected marine organisms. Principal component analysis revealed the exemplary antifouling efficacy of the sponge extracts of Stylissa sp. observed followed by Hyrtios sp. against bacterial forms in the laboratory study. De-trended correspondence analysis confirmed that the contribution of antifouling efficacy of the selected sponge extracts was observed to be more towards Bacillus sp., Vibrio sp. and Alteromonas sp. Moreover, the efficacy of Hyrtios sp. extract(20.430 μg m L~(-1)) followed by Stylissa sp.(30.945 μg m L~(-1)) showed higher against barnacle instar compared with other extracts in the bioactivity assay. Bray-Curtis cluster analysis under paired linkage categorized all the sponge extracts into one major cluster with 75% similarity, and one outlier tunicate. More than 80% similarity observed between Hyrtios sp. and Stylissa sp. Fourier transform infrared spectroscopy(FTIR) showed that the contribution of major peaks found in the marine organisms were towards sulfones, sulfoxides, cyanates and ketones.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.     

Development and evaluation of a DNA microarray assay for the simultaneous detection of nine harmful algal species in ship ballast and seaport waters

Rapid, high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae, which are responsible for algal blooms, such as red and green tides. In this study, we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously, namely Alexandrium tamarense, Gyrodinium instriatum, Heterosigma akashiwo, Karenia mikimotoi, Prorocentrum donghaiense, Prorocentrum minimum, Ulva compressa, Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection (LOD) of 0.5 ng of genomic DNA (orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether, 230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%, respectively, relative to conventional morphological methods. This indicated that this high-throughput, automatic, and specific method is well suited for the detection of algae in water samples.     

Seasonal variation and principle of cyanobacterial biomass and forms in the water source area of Chaohu City,China

We investigated seasonal variations in cyanobacterial biomass and the forms of its dominant population(M. aeruginosa) and their correlation with environmental factors in the water source area of Chaohu City,China from December 2011 to October 2012. The results show that species belonging to the phylum Cyanophyta occupied the maximum proportion of phytoplankton biomass,and that the dominant population in the water source area of Chaohu City was M. aeruginosa. The variation in cyanobacterial biomass from March to August 2012 was well fitted to the logistic growth model. The growth rate of cyanobacteria was the highest in June,and the biomass of cyanobacteria reached a maximum in August. From February to March 2012,the main form of M.aeruginosa was the single-cell form; M.aeruginosa colonies began to appear from April,and blooms appeared on the water surface in May. The maximum diameter of the colonies was recorded in July,and then gradually decreased from August. The diameter range of M. aeruginosa colonies was 18.37–237.77 μm,and most of the colonies were distributed in the range 20–200 μm,comprising 95.5% of the total number of samples. Temperature and photosynthetically active radiation may be the most important factors that influenced the annual variation in M. aeruginosa biomass and forms. The suitable temperature for cyanobacterial growth was in the range of 15–30°C. In natural water bodies,photosynthetically active radiation had a significant positive influence on the colonial diameter of M.aeruginosa(P0.01).     

Characterization and in-vivo evaluation of potential probiotics of the bacterial flora within the water column of a healthy shrimp larviculture system

A thorough understanding of the normal bacterial flora associated with shrimp larviculture systems contributes to probiotic screening and disease control.The bacterial community of the water column over a commercial Litopenaeus vannamei larval rearing run was characterized with both culture-dependent and culture-independent methods.A total of 27 phylotypes at the species level were isolated and identified based on 16 S rDNA sequence analysis.Denaturing gradient gel electrophoresis(DGGE) analysis of the V3–V5 region of 16 S rRNA genes showed a dynamic bacterial community with major changes occurred from stages zoea to mysis during the rearing run.The sequences retrieved were affiliated to four phyla,Proteobacteria,Actinobacteria,Bacteroidetes,and Firmicutes,with the family Rhodobacteraceae being the most frequently recovered one.Subsequently,13 representative strains conferred higher larval survival than the control when evaluated in the in-vivo experiments;in particular,three candidates,assigned to Phaeobacter sp.,Arthrobacter sp.,and Microbacterium sp.,significantly improved larval survival( P 0.05).Therefore,the healthy shrimp larviculture system harbored a diverse and favorable bacterial flora,which contribute to larval development and are of great importance in exploiting novel probiotics.     

Karyotype analysis of four nemertean species

We performed a karyotype analysis on four species from Phylum Nemertea using regenerating somatic cells. Two palaeonemertean species, Cephalothrix hongkongiensis and Cephalothrix sp., had the same chromosome number (2n=28), but different karyotypes (16 m+10 sm+2 st, NF=54 and 22 m+4 sm+2 st, NF=54, respectively). The karyotypes of the two heteronemerteans, Notospermus geniculatus and Ramphogordius sanguineus were 2n=38, 30 m+6 sm+2 sm/st (NF=76/74) and 2n=56, 48 m+4 sm+4 sm/st (NF=112/108), respectively.     

Impact of <Emphasis Type="Italic">Vibrio parahaemolyticus</Emphasis> and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>

White spot syndrome virus (WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect Litopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus (WSSV+Vp), or we first infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection (Vp+WSSV). The eff ect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of Vibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction (PCR) method. LvMyD88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the diff erent mortality rates. Our results show that (1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection; (2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed significant increases in the numbers of Vibrio (P<0.05); (3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10⁵ to 10⁷ /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.     

Morphological study of the genus <Emphasis Type="Italic">Herposiphonia</Emphasis> (Rhodophyta,Rhodomelaceae) on the coast of eastern Guangdong,China, with a description of <Emphasis Type="Italic">H. pinnata</Emphasis> sp. nov.

We present a taxonomic study of taxa of the red algae genus Herposiphonia (Rhodophyta, Rhodomelaceae), collected from the coast of eastern Guangdong, China. We made detailed morphological studies and considered recent taxonomic criteria for species delimitation, and are making the first report of five different species on the coast of Guangdong, including a new species. The species identified were H. caespitosa Tseng, H. hollenbergii Dawson, H. pecten - veneris (Harvey) Falkenberg, H. subdisticha Okamura and H. pinnata Ding and Tan sp. nov. H. pinnata sp. nov. is characterized by bright green thalli; most parts of the feathery thalli are free of the substratum; determinate branches and indeterminate branches are arranged in a chaotic sequence; the primary axis has bare segments; the determinate branch has 9–11 periaxial cells per segment; vegetative trichoblasts are abundant; and tetrasporangia are formed on the middle of the determinate branch with 1–8 successive segments in a single rectilinear series. This paper is also the first record of sporophyte plants of H. pecten-veneris.     

Characterization of proteases from <Emphasis Type="Italic">Planomicrobium</Emphasis> sp. L-2 isolated from the gastrointestinal tract of <Emphasis Type="Italic">Octopus variabilis</Emphasis> (Sasaki)

A crude protease produced from Planomicrobium sp. L-2 is described, and its effectiveness as an additive in liquid detergent evaluated. We isolate the protease-producing Planomicrobium sp. L-2 from the gastrointestinal tract of Octopus variabilis. At least three caseinolytic protease clear bands were observed in zymogram analysis. The crude alkaline protease was highly tolerant of a pH range from 7.0 to 9.0, and temperatures to 50°C after incubation for 1 h. Proteolytic enzymes were stable towards three surfactants (5% Tween 80, 1% Triton X-100 and 0.05% SDS) and an oxidizing agent (1% hydrogen peroxide), in addition to being highly stable and compatible with popular commercial laundry powered detergent brands available in China. Our study demonstrates the potential these proteases have for development into novel classes of detergent additive. This study also suggests that the gastrointestinal tract of Octopus variabilis may be a rich source of commercially valuable strains of enzyme.     

Two new free-living marine nematode species of the genus Anoplostoma (Anoplostomatidae) from the mangrove habitats of Xiamen Bay,East China Sea

Two new species of free-living marine nematode from mangrove habitats in Xiamen Bay are identified.Anoplostoma tumidum sp.nov.is characterized by relatively short outer labial setae(0.86–1.00 h.d.),long tail(c 7.2–8.9,c' 8.3–10.5),an instinct swollen distal portion of slender spicule(Sc 94–101 μm),and well developed copulatory bursae without bursal papillae.A.tumidum sp.nov.differs from all valid species of genus Anoplostoma in copulatory apparatus of males with a distinct swollen distal portion of spicule,and a relatively obvious constriction of head.A.paraviviparum sp.nov.is characterized by relatively long outer labial setae(1.11–1.22 h.d.),and tail(c 6.6–8.5,c' 8.6–10.2);elongated spicules with distinct knob-like proximal and pointed distal ends(Sc 46–69 μm);distinct strip-like gubernaculum(length with 11–15μm);well developed copulatory bursae with precloacal papillae and post-cloacal papillae;and a distinct constriction of head.A.paraviviparum sp.nov.is similar to A.viviparum Bastian,1865,but differs in the reproductive mode of female and the constriction of head.     

Morphology and SSU rDNA sequence analysis of two hypotrichous ciliates (Protozoa,Ciliophora, Hypotrichia) including the new species <Emphasis Type="Italic">Metaurostylopsis parastruederkypkeae</Emphasis> n. sp.

The morphology and phylogeny of two hypotrichous ciliates, Metaurostylopsis parastruederkypkeae n. sp. and Neourostylopsis flavicana (Wang et al., 2011) Chen et al., 2013 were investigated based on morphology, infraciliature and the small subunit (SSU) ribosomal RNA gene (rRNA) sequence. The new species, M. parastruederkypkeae n. sp. was identified according to its characteristics: body shape ellipsoidal, size about (165–200) × (45–60) μm in vivo, cell color reddish; two types of cortical granules including wheat grain-like and yellow-greenish larger ones along the marginal cirri rows and dorsal kineties and dot-like and reddish smaller ones, grouped around marginal cirri on ventral side and arranged in short lines on dorsal side; 26–41 adoral membranelles; three frontal and one parabuccal, five to seven frontoterminal, one buccal, and three to six transverse cirri; seven to thirteen midventral pairs; five to nine unpaired ventral cirri, five to seven left and three to five right marginal rows; and three complete dorsal kineties. Phylogenetic analysis based on SSU rDNA sequences showed that both Metaurostylopsis and Neourostylopsis are monophyletic. As the internal relationship between and within both genera are not clear, further studies on the species in these two genera are necessary. The key characteristics of all known twelve Metaurostylopsis-Apourostylopsis-Neourostylopsis species complex were updated.     

The mycobiota of discoloured needles of the Swiss stone pine (<Emphasis Type="Italic">Pinus cembra</Emphasis> L.) in the Tatra Mountains (Poland)

The Swiss stone Pine (Pinus cembra L.) is an alpine species, fairly commonly occurring in the Alps and the Carpathians, close to the timberline. Natural sites of the Swiss stone pine in Poland are found exclusively in the Tatra Mountains, within the area of the Tatra National Park (TNP). In 2017, the health status of the needles of P. cembra was determined and their mycological analysis was carried out. As a result, 11 species of fungi were isolated from the needles showing disease symptoms, manifested by various shades of discoloration. The most frequently observed species was Fusarium oxysporum. It probably does not bring on disease symptoms, but may affect the development of other frequently isolated species (such as Sydowia polyspora, Lophodermium sp. and Lophodermium conigenum) that are pathogenic to Pinus spp.     

Morphology and controlling factors of the longitudinal profile of gullies in the Yuanmou dry-hot valley

The morphology of the gully longitudinal profile (GLP) is an important topographic index of the gully bottom associated with the evolution of the gullies. This index can be used to predict the development trend and evaluate the eroded volumes and soil losses by gullying. To depict the morphology of GLP and understand its controlling factors, the Global Positioning System Real-time Kinematic (GPS RTK) and the total station were used to measure the detail points along the gully bottom of 122 gullies at six sites of the Yuanmou dry-hot Valley. Then, nine parameters including length (Lt), horizontal distance (Dh), height (H), vertical erosional area (A), vertical curvature (Cv), concavity (Ca), average gradient (Ga), gully length-gradient index (GL), normalized gully length-gradient index (Ngl), were calculated and mapped using CASS, Excel and SPSS. The results showed that this study area is dominated by slightly concave and medium gradient GLPs, and the lithology of most gullies is sandstone and siltstone. Although different types of GLPs appear at different sites, all parameters present a positively skewed distribution. There are relatively strong correlations between several parameters: namely Lt and H, Dh and H, Lt and A, Dh and A, H and GL. Most GLPs, except three, have a best fit of exponential functions with quasistraight shapes. Soil properties, vegetation coverage, piping erosion and topography are important factors to affect the GLP morphology. This study provides useful insight into the knowledge of GLP morphology and its influential factors that are of critical importance to prevent and control gully erosion.     

Taxonomy,Morphology and Phylogeny of a New Oligotrich Ciliate–<Emphasis Type="Italic">Omegastrombidium hongkongense</Emphasis> n. sp. (Protozoa: Ciliophora) from Clear Water Bay,Hong Kong

One new marine oligotrich ciliate, Omegastrombidium hongkongense n. sp., was isolated from a bloom of Noctiluca scuntillans near Port Shelter, Hong Kong. The morphology and infraciliature of this new species were studied on both living and protargol-stained specimens. Its phylogenetic position was discussed based on the sequence of the small subunit rRNA gene. O. hongkongense is different from its congeners with special characters. The cells are usually heart-shaped, and the cell size usually is (20–35) × (20–30) μm in vivo. Its deep buccal cavity extends obliquely to about 1/2 of cell length. It shows prominent apical protrusion. The adoral zone of membranelles is divided into 17–19 collar membranelles and four buccal membranelles. It has one ball-like macronucleus. The girdle kinety forms a closed loop which obliquely surrounds the body. The ventral kinety and thigmotactic membranelles are not observed. The SSU rRNA sequence of O. hongkongense was close to those of Strombidium paracalkinsi and Varistrombidium kelum with approximately 99% similarity. In the phylogenetic trees, O. hongkongense can be grouped with O. elegans and V. kielum species with very low support (16% ML).     

Characterization and identification of enzyme-producing microflora isolated from the gut of sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Gut microorganisms play an important role in the digestion of their host animals. The purpose of this research was to isolate and assess the enzyme-producing microbes from the Apostichopus japonicus gut. Thirty-nine strains that can produce at least one of the three digestive enzymes (protease, amylase, and cellulase) were qualitatively screened based on their extracellular enzyme-producing abilities. The enzyme-producing strains clustered into eight groups at the genetic similarity level of 100% by analyzing the restriction patterns of 16S rDNA amplified with Mbo I. Phylogenetic analysis revealed that 37 strains belonged to the genus Bacillus and two were members of the genus Virgibacillus. Enzyme-producing capability results indicate that the main enzyme-producing microflora in the A. japonicus gut was Bacillus, which can produce protease, amylase, and cellulase. Virgibacillus, however, can only produce protease. The high enzyme-producing capability of the isolates suggests that the gut microbiota play an important role in the sea cucumber digestive process.     

<Emphasis Type="Italic">Grateloupia ramosa</Emphasis> Wang &amp; Luan sp. nov. (Halymeniaceae,Rhodophyta), a new species from China based on morphological evidence and comparative <Emphasis Type="Italic">rbc</Emphasis>L sequences

Grateloupia ramosa Wang & Luan sp. nov. (Halymeniaceae, Rhodophyta) is newly described from Hainan Province, southern China. The organism has the following morphological features: (1) purplish red, cartilaginous and lubricous thalli 5–10 cm in height; (2) compressed percurrent axes bearing abundant branches with opposite arrangement; (3) claw-like apices on top, constricted to 2–4 cm at the base; (4) cortex consisting of 3–6 layers of elliptical or anomalous cells and a medulla covered by compact medullary filaments; (5) reproductive structures distributed throughout the thallus, especially centralized at the bottom of the end portion of the branches; and (6) 4-celled Carpogonial branches and 3-celled auxiliary-cell branches, both of the Grateloupia-type. The morphological diff erences were supported by molecular phylogenetics based on ribulose-1, 5-bisphosphate carboxylase/oxygenase (rbcL) gene sequence analysis. There was only a 1 bp divergence between specimens collected from Wenchang and Lingshui of Hainan province. The new species was embedded in the large Grateloupia clade of the Halymeniaceae. The pairwise distances between G. ramosa and other species within Grateloupia ranged from 26 to 105 bp, within pairwise distances of 13–111 bp between species of the large genus Grateloupia in Halymeniaceae. Thus, we propose this new species as G. ramosa Wang & Luan sp. nov.     

Succession and seasonal variation in epilithic biofilms on artificial reefs in culture waters of the sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

Periphytic biofilms in aquaculture waters are thought to improve water quality, provide an additional food source, and improve the survival and growth of some reared animals. In the Asia- Pacific region, particularly in China, artificial reefs are commonly used in the commercial farming of sea cucumbers. However, few studies have examined the epilithic biofilms on the artificial reefs. To gain a better understanding of the succession of epilithic biofilms and their ecological processes in sea cucumber culture waters, two experiments were conducted in culture waters of the sea cucumber Apostichopus japonicus in Rongcheng, China, using artificial test panels. On the test panels of succession experiment, more than 67 species were identified in the biofilms. On the test panels of seasonal variation experiment, more than 46 species were recorded in the biofilms. In both experiments, communities of epilithic biofilms were dominated by diatoms, green algae and the annelid Spirorbis sp. In the initial colonization, the dominant diatoms were Cocconeis sp., Amphora spp. and Nitzschia closterium in June, which were succeeded by species of Navicula, Cocconeis and Nitzschia (July to September), and then by Licmophora abbreviata, Nitzschia closterium and Synedra spp. in the following months. A diatom bloom in the autumn and filamentous green algae burst in the summer were also observed. Ecological indices well annotated the succession and seasonal changes in epilithic communities. Multidimensional scaling (MDS) analysis found significant differences in diatom community composition among months and seasons. Fast growth of biofilms was observed in the summer and autumn, whereas the biomass of summer biofilms was largely made up of filamentous green algae. Present results show that the components of epilithic biofilms are mostly optimal foods of A. japonicus, suggesting that biofilms on artificial reefs may contribute important nutritional sources for sea cucumbers during their growth seasons. Future works should include quantitative determination of the contribution of epilithic biofilms to the diet of A. japonicus, potential roles of epilithic biofilms in regulating the water quality of sea cucumber ponds, and the regulation of epilithic biofilms in sea cucumber culture ponds.