Establishment and characterization of a testicular Sertoli cell line from olive flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

The culture of Sertoli cells has become an indispensable resource in studying spermatogenesis. A new Sertoli cell line (POSC) that consisted predominantly of fibroblast-like cells was derived from the testis of the olive flounder Paralichthys olivaceus and sub-cultured for 48 passages. Analysis of the mtDNA COI gene partial sequence confirmed that the cell line was from P. olivaceus. Cells were optimally maintained at 25°C in DMEM/F12 medium supplemented with fetal bovine serum, basic fibroblast growth factor, and epidermal growth factor. The growth curve of POSC showed a typical “S” shape. Chromosome analysis revealed that the cell line possessed the normal P. olivaceus diploid karyotype of 2n=48t. POSC expressed dmrt1 but not vasa, which was detected using RT-PCR and sequencing. Immunocytochemistry revealed that the cells exhibited the testicular Sertoli cell marker FasL. Therefore, POSC appeared to consist of testicular Sertoli cells. Bright fluorescent signals were observed after the cells were transfected with pEGFP-N3 plasmid, with the transfection efficiency reaching 10%. This research not only offers an ideal model for further gene expression and regulation studies on P. olivaceus, but also serves as valuable material in studying fish spermatogenesis, Sertoli cell-germ cell interactions, and the mechanism of growth and development of testis.     

Effect of twine thickness on size-selectivity of driftnet for the yellow croaker <Emphasis Type="Italic">Larimichthys polyactis</Emphasis> in southwestern Sea of Korea

In order to determine the effect of twine thickness on the size-selectivity of the driftnet used for the yellow croaker, size-selectivity tests were conducted with three different twine thicknesses (monofilament diameters of 0.279 mm (number’s method; No. 3), 0.321 mm (No. 4), and 0.360 mm (No. 5)) of driftnets for the yellow croaker in the seas around Chooja-do, Jeju Islands. The selectivity curve was estimated by using Kitahara’s method. In order to determine the physical properties of the twine used in the experimental fishing nets, we measured the breaking load, elongation, and stiffness under both dry and wet conditions. In terms of physical properties, the thinnest twine (No. 3) had the strongest breaking strength per unit cross-sectional area, along with good elongation and excellent flexibility. The thickest twine (No. 5) had the lowest flexibility. In terms of selectivity, the net of No. 3 twine showed the broadest selection range and, thus, a relatively low selectivity compared with the other nets, while the less flexible net of No. 5 twine showed the narrowest selectivity range and high selectivity. In addition, it was found that a thicker twine resulted in a smaller haul of small fish. Therefore, it can be inferred that the thickness of the twine affects the size of the catch and selectivity, and thus the size composition of the catch as well.     

Impact factors identification of spatial heterogeneity of herbaceous plant diversity on five southern islands of Miaodao Archipelago in North China

Herbaceous plants are widely distributed on islands and where they exhibit spatial heterogeneity. Accurately identifying the impact factors that drive spatial heterogeneity can reveal typical island biodiversity patterns. Five southern islands in the Miaodao Archipelago, North China were studied herein. The spatial distribution of herbaceous plant diversity on these islands was analyzed, and the impact factors and their degree of impact on spatial heterogeneity were identified using CCA ordination and ANOVA. The results reveal 114 herbaceous plant species, belonging to 94 genera from 34 families in the 50 plots sampled. The total species numbers on different islands were significantly positively correlated with island area, and the average α diversity was correlated with human activities, while the β diversity among islands was more affected by island area than mutual distances. Spatial heterogeneity within islands indicated that the diversities were generally high in areas with higher altitude, slope, total nitrogen, total carbon, and canopy density, and lower moisture content, pH, total phosphorus, total potassium, and aspect. Among the environmental factors, pH, canopy density, total K, total P, moisture content, altitude, and slope had significant gross effects, but only canopy density exhibited a significant net effect. Terrain affected diversity by restricting plantation, plantation in turn influenced soil properties and the two together affected diversity. Therefore, plantation was ultimately the fundamental driving factor for spatial heterogeneity in herbaceous plant diversity on the five islands.     

Contribution of phytoliths to total biogenic silica volumes in the tropical rivers of Malaysia and associated implications for the marine biogeochemical cycle

The contribution of phytoliths to total biogenic silica (BSi) volumes in rivers worldwide, and the associated implications for the biogeochemical cycle, require in-depth study. Based on samples from rivers in Peninsular Malaysia, this project investigated the source and characteristics of BSi found in Asian tropical rivers, as well as the process of reverse weathering taking place in these fluvial systems. Results indicated that BSi samples collected in sediments consisted of phytolith, diatom and sponge spicules. Phytoliths, predominantly of the elongate form, comprised 92.8%–98.3% of BSi in the Pahang River. Diatom BSi in this river consisted mainly of pennatae diatoms, but represented a relatively small proportion of the total BSi volume. However, diatom BSi (predominantly of the Centricae form) was more prevalent in the Pontian and Endau Rivers with shares of 68.8% and 79.3% of the total BSi volumes, respectively, than Pahang River. Carbon contents of the BSi particulates ranged from 1.85% to 10.8% with an average of 4.79%. These values are higher than those recorded in other studies to date, and indicate that BSi plays a major role in controlling permanent carbon burial. This study suggests that phytoliths from terrestrial plants are the primary constituents of BSi in the rivers of Peninsular Malaysia, and therefore represent a significant proportion of the coastal silica budget.     

Significant population genetic structure detected in the rock bream <Emphasis Type="Italic">Oplegnathus fasciatus</Emphasis> (Temminck &amp; Schlegel, 1844) inferred from fluorescent-AFLP analysis

Oplegnathus fasciatus (rock bream) is a commercial rocky reef fish species in East Asia that has been considered for aquaculture. We estimated the population genetic diversity and population structure of the species along the coastal waters of China using fluorescent-amplified fragment length polymorphisms technology. Using 53 individuals from three populations and four pairs of selective primers, we amplified 1 264 bands, 98.73% of which were polymorphic. The Zhoushan population showed the highest Nei’s genetic diversity and Shannon genetic diversity. The results of analysis of molecular variance (AMOVA) showed that 59.55% of genetic variation existed among populations and 40.45% occurred within populations, which indicated that a significant population genetic structure existed in the species. The pairwise fixation index F _st ranged from 0.20 to 0.63 and were significant after sequential Bonferroni correction. The topology of an unweighted pair group method with arithmetic mean tree showed two significant genealogical branches corresponding to the sampling locations of North and South China. The AMOVA and STRUCTURE analyses suggested that the O. fasciatus populations examined should comprise two stocks.     

An integrative study of larval organogenesis of American shad <Emphasis Type="Italic">Alosa sapidissima</Emphasis> in histological aspects

We describe organogenesis at a histological level in American shad (Alosa sapidissima) larvae from 0 until 45 days after hatching (DAH). Larval development was divided into four stages based on the feeding mode, external morphological features, and structural changes in the organs: stage 1 (0–2 DAH), stage 2 (3–5 DAH), stage 3 (6–26 DAH) and stage 4 (27–45 DAH). At early stage 2 (3 DAH), American shad larvae developed the initial digestive and absorptive tissues, including the mouth and anal opening, buccopharyngeal cavity, oesophagus, incipient stomach, anterior and posterior intestine, differentiated hepatocytes, and exocrine pancreas. The digestive and absorptive capacity developed further in stages 2 to 3, at which time the pharyngeal teeth, taste buds, gut mucosa folds, differentiated stomach, and gastric glands could be observed. Four defined compartments were discernible in the heart at 4 DAH. From 3 to 13 DAH, the excretory systems started to develop, accompanied by urinary bladder opening, the appearance and development of primordial pronephros, and the proliferation and convolution of renal tubules. Primordial gills were detected at 2 DAH, the pseudobranch was visible at 6 DAH, and the filaments and lamellae proliferated rapidly during stage 3. The primordial swim bladder was first observed at 2 DAH and started to inflate at 9 DAH; from then on, it expanded constantly. The spleen was first observed at 8 DAH and the thymus was evident at 12 DAH. From stage 4 onwards, most organs essentially manifested an increase in size, number, and complexity of tissue structure.     

Heavy metals in sediments and their bioaccumulation in <Emphasis Type="Italic">Phragmites australis</Emphasis> in the Anzali wetland of Iran

Accumulation of metals in both sediments and Phragmites australis organs was studied. Samples were collected from seven stations located in Anzali wetland, Iran. The samples were analyzed by inductively coupled plasma mass spectrometry (ICP-MS). The results showed that concentration of the studied metals (except As and Cd) were higher in sediments than in P. australis organs. Metal accumulation was found to be significantly (P <0.05) higher in roots than in above-ground organs of P. australis. The bioaccumulation factor (BAF) and the transfer factor (TF) also verified the highest rate of metal accumulation in roots and their reduced mobility from roots to the above-ground organs. Pearson correlation coefficient showed significant relationships between metal concentrations in sediments and those in plant organs. It should be pointed out that sediment and plant samples exhibited higher metal concentrations in eastern and central parts than in western and southern parts of the wetland. The mean concentrations of all studied elements (except for Fe, V and Al) were higher in these sediment samples than in the Earth’s crust and shale. High accumulation of metals in P. australis organs (roots and shoots) is indicative of their high bioavailability in sediments of the wetland. The correlation between metal concentrations in sediments and in P. australis indicates that plant organs are good bioindicators of metal pollution in sediments of Anzali wetland.     

Phylogenetic diversity of culturable bacteria in surface seawater from the Drake Passage,Antarctica

The Drake Passage is located between the Antarctic Peninsula and Tierra del Fuego in the south of South America. Surface seawater samples were collected at seven sites in the Drake Passage during the austral summer of 2012. The 16S rRNA sequences were analyzed from 187 isolated bacterial strains. Three phyla, 29 genera and 56 species were identified. The three phyla were Actinobacteria, Firmicutes and Proteobacteria; the Proteobacteria included α-Proteobacteria, β-Proteobacteria and γ-Proteobacteria. γ-Proteobacteria, Actinobacteria and Firmicutes were the dominant class or phyla in terms of quantity and species. Gram-positive bacteria (Actinobacteria and Firmicutes) accounted for 57.8% of all types identified. There were nine dominant genera, including Curtobacterium, Staphylococcus, and Halomonas, and 14 dominant species including Curtobacterium flaccumfaciens, Curtobacterium pusillum, and Staphylococcus sciuri. Of the strains identified, 87.2% were catalase positive or weakly positive.     

江西牛形坝-柳木坑金银铅锌多金属矿床铋矿物类型及指示意义

江西银坑矿田的牛形坝-柳木坑矿床是赣南于都-赣县矿集区典型的金银铅锌多金属矿床。根据野外调研、系统矿相学研究和电子探针成分分析,表明铋在牛形坝-柳木坑矿床中是一种重要的伴生矿化金属元素,呈独立矿物或呈微细矿物包体分布在早期硫化物内部或边缘。铋矿物种类丰富,主要包括块硫铋银矿、硫铋银矿、板硫铋铜铅矿、针硫铋铅矿、硫铜铋矿、库辉铋铜铅矿、辉碲铋矿等。与同一矿集区石英脉型盘古山钨铋矿床相比,两矿床铋矿物种类均出现Pb-Ag-Bi-S、Te-Bi-S元素组合,赋存形式和含量有所不同,Cu-Pb-Bi-S元素组合仅在牛形坝-柳木坑矿床出现。矿床属于Au-Bi-(低Te)-S体系,铋盐矿物形成于中低温、中高密度、低盐度的成矿环境。从岩浆中分馏出来的成矿流体发生减压沸腾迫使大量蒸汽相逸散,硫逸度增加,铋主要以硫络合物形式运移,随着黄铁矿等硫化物沉淀,硫被大量消耗形成金矿物和碲化物(碲银矿)。成矿流体演化过程中由于温度和硫逸度进一步降低导致在成矿后期沉淀出大量铋硫盐矿物。成矿过程中的Bi促进金的迁移富集。铋矿物在两矿床中的赋存特征、化学成分、形成环境以及沉淀机制等表现出差异,说明铋具有成矿选择性,起到对成矿物质示踪的作用。     

Source and yearly distribution of PAHs in the snow from the Hailuogou glacier of Mountain Gongga,China

Snow samples were collected over a 3-year period from 2012 to 2014 at the Hailuogou glacier of Mountain Gongga (Mt. Gongga) and analyzed for 16 priority polycyclic aromatic hydrocarbons (PAHs) using Gas Chromatography–Mass Spectrometry (GC–MS). The results show that total average levels of the 16 PAHs ranged from 452 to 290 ng·L^?1 with a possible declining trend from 2012 to 2014. Distances between the sampling sites and the emission sources were estimated at typically less than 500 km. The results suggest that the major source of PAHs was from coal combustion, while contributions from automobile exhaust played an important role in more recent years. This finding was in agreement with the characteristics of presence of local industry, residence, and recent development of tourism of the surrounding areas.