Altitudinal trends in δ~(13)C value,stomatal density and nitrogen content of Pinus tabuliformis needles on the southern slope of the middle Qinling Mountains,China

In this study,a coniferous tree species(Pinus tabuliformis Carr.) was investigated at a moderate-altitude mountainous terrain on the southern slope of the middle Qinling Mountains(QLM) to detect the trends in carbon isotope ratio( δ~(13)C),leaf nitrogen content(LNC) and stomatal density(SD) with altitude variation in northsubtropical humid mountain climate zone of China.The results showed that LNC and SD both significantly increased linearly along the altitudinal gradient ranging from 1000 to 2200 m,whereas leafδ~(13)C exhibited a significantly negative correlation with the altitude.Such a correlation pattern differs obviously from that obtained in offshore low-altitude humid environment or inland high-altitude semi-arid environment,suggesting that the pattern of increasing δ~(13)C with the altitude cannot be generalized.The negative correlation between δ ~(13)C and altitude might be attributed mainly to the strengthening of carbon isotope fractionation in plants caused by increasing precipitation with altitude.Furthermore,there was a remarkable negative correlation between leaf δ ~(13)C and LNC.One possible reason was that increasing precipitation that operates to increase isotopic discrimination with altitude overtook the LNC in determining the sign of leaf δ ~(13)C.The significant negative correlation between leaf δ ~(13)C and SD over altitudes was also found in the present study,indicating that increases in SD with altitude would reduce,rather than enhance plant δ~(13)C values.     

Peat humification- and δ<Superscript>13</Superscript>C<Subscript>cellulose</Subscript>-recorded warm-season moisture variations during the past 500 years in the southern Altai Mountains within northern Xinjiang of China

To predict future spatio-temporal patterns of climate change, we should fully understand the spatio-temporal patterns of climate change during the past millennium. But, we are not yet able to delineate the patterns because the qualities of the retrieved proxy records and the spatial coverage of those records are not adequate. Northern Xinjiang of China is one of such areas where the records are not adequate. Here, we present a 500-yr land-surface moisture sequence from Heiyangpo Peat (48.34°N, 87.18°E, 1353 m a.s.l) in the southern Altai Mountains within northern Xinjiang. Specifically, peat carbon isotope value of cellulose (δ¹³C_cellulose) was used to estimate the warm-season moisture variations and the degree of humification was used to constrain the δ¹³C_cellulose-based hydrological interpretation. The climatic attributions of the interpreted hydrological variations were based on the warm-season temperature reconstructed from Belukha ice core and the warm-season precipitation inferred from the reconstructed Atlantic Multidecadal Oscillations (AMO). The results show that humification decreased and the δ¹³C_cellulose-suggested moisture decreased from ~1510 to ~1775 AD, implying that a constant drying condition may have inhibited peat decay. Our comparison with reconstructed climatic parameters suggests that the moisture-level decline was most likely resulted from a constant decline of precipitation. The results also show that humification kept a stable level and the δ¹³C_cellulose-suggested moisture also decreased from ~1775 to ~2013 AD, implying that peat decay in the acrotelm primarily did not depend on the water availability or an aerobic environment. Again, our comparison with reconstructed climatic parameters suggests that the land-surface moisturelevel decline was most likely resulted from a steady warming of growing-season temperature.     

Exploration of Antifouling Potential of the Brown Algae <Emphasis Type="Italic">Laminaria</Emphasis> ‘Sanhai’

Seaweeds are one of the largest producers of biomass in the marine environment. It has been well known that marine algae, especially brown algae was a rich source of biogenic compounds with antifouling potential that could be ideal alternatives of tributyltin (TBT). In this paper, antifouling potential of the brown algae Laminaria ‘sanhai’ was explored. Firstly, the dried alga was extracted and the antialgal and antilarval activities were investigated. The EC₅₀ and LC₅₀ values of crude extract of Laminaria ‘sanhai’ against diatom (Skeletonema costatum) and barnacle larval (Chthamalus challengeri) were 8.9 μg mL^?1 and 12.0 μg mL^?1 respectively. Then, guided by bioassay, the bioactive substances were isolated by liquid-liquid extraction. The antialgal and antilarval activities of isolated fraction were improved with the EC₅₀ value of 7.4 μg mL^?1 against S. costatum and LC₅₀ value of 9.7 μg mL^?1 against C. challengeri larvae. Identification by IR, Q-TOFMS and GC-MS of the isolated bioactive substances revealed the abundance of fatty acids. These fatty acids, most with 16, 18 or 20 carbon atoms, contained myristic, hexadecanoic, oleic, linolenic, arachidonic and eicosapentaenoic acids. The results indicated that both the crude extract and the isolated bioactive substances had high antialgal and antilarval activities with no highlighted cytotoxicity which made the brown algae Laminaria ‘sanhai’ a promising source of the environmentally friendly antifoulants.     

Isolation and characterization of pathogenic <Emphasis Type="Italic">Listonella anguillarum</Emphasis> of diseased half-smooth tongue sole (<Emphasis Type="Italic">Cynoglossus semilaevis</Emphasis> Günther)

Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD₅₀ value of <1.0×10⁵ colony forming units (CFU) mL⁻¹, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillarum with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.     

Designation of a new genus <Emphasis Type="Italic">Bathymenes</Emphasis> for the deep-sea pontoniine shrimps of the ‘<Emphasis Type="Italic">Periclimenes alcocki</Emphasis> species group’ (Decapoda,Caridea, Palaemonidae), with a checklist of the species assigned to the genus

A new genus of the deep-sea pontoniine shrimps, Bathymenes gen. nov., is established for the ‘Periclimenes alcocki species group’ of the genus Periclimenes Costa, 1844. The new genus is distinguished from other genera of the Pontoniinae by a combination of characters: the posteriormost dorsal rostral tooth remote from other teeth, situated in the epigastric position, cornea usually reduced, the propodus of second pereiopods covered with fine granules, the dactylus of the major second chela being generally flanged, the ambulatory pereiopods with the dactyli being biunguiculate and telson with more than two pairs of dorsolateral spines. The genus is mainly distributed in the tropical to warm-temperate Indo-West Pacific waters at depths greater than 200 m. Fifteen species previously recognized as belonging to the ‘P. a lcocki species group’ are now placed in Bathymenes gen. nov. A key for their identification and a checklist of congeneric species are provided.     

Isolation and characterization of bioactive fungi from shark <Emphasis Type="Italic">Carcharodon carcharias</Emphasis>’ gill with biopharmaceutical prospects

Until recently, little was known about the fungi found in shark gills and their biomedicinal potential. In this article, we described the isolation, bioactivity, diversity, and secondary metabolites of bioactive fungi from the gill of a shark (Carcharodon carcharias). A total of 115 isolates were obtained and grown in 12 culture media. Fifty-eight of these isolates demonstrated significant activity in four antimicrobial, pesticidal, and cytotoxic bioassay models. Four randomly selected bioactive isolates inhibited human cancer cell proliferation during re-screening. These active isolates were segregated into 6 genera using the internal transcribed spacer-large subunit (ITS-LSU) rDNA-sequence BLAST comparison. Four genera, Penicillium, Aspergillus, Mucor, and Chaetomium were the dominant taxa. A phylogenic tree illustrated their intergenera and intragenera genetic diversity. HPLC-DAD-HRMS analysis and subsequent database searching revealed that nine representative strains produced diverse bioactive compound profiles. These results detail the broad range of bioactive fungi found in a shark’s gills, revealing their biopharmaceutical potential. To the best of our knowledge, this is the first study characterizing shark gill fungi and their bioactivity.     

Effects of dietary β-glucan and glycyrrhizin on non-specific immunity and disease resistance of the sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka) challenged with <Emphasis Type="Italic">Vibrio splendidus</Emphasis>

Sea cucumbers, Apostichopus japonicus Selenka, were fed diets containing non-immunostimulant (basal diet), 0.2% β-glucan and 0.02% glycyrrhizin in a recirculatory water system for 45 days, and subsequently challenged with Vibrio splendidus by injection at 1.0×10⁸ cfu / sea cucumber for 15 days. Phagocytic capacity (PC), intracellular superoxide anion production (ISAP), lysozyme (LSZ) activity and superoxide dismutase (SOD) activity in the coelomic fluid were analyzed on the 0^th, 5^th, 10^th and 15^th days after injection. Results showed that after the 45-day feeding period, PC, ISAP, LSZ activity and SOD activity in sea cucumbers fed with dietary β-glucan or glycyrrhizin were significantly higher than in those fed with the basal diet. On the 5th day after infection, all the immune parameters examined in the sea cucumbers injected with V. splendidus decreased in value significantly. On the 15^th day, PC, ISAP and LSZ activity returned to levels similar to those on the 0^th day. For the sea cucumbers injected with saline, there were no significant differences in all the immune parameters examined and in the cumulative morbidity during the 15-day challenging trial. After injecting with V. splendidus, the cumulative morbidity of sea cucumbers fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin when challenged with V. splendidus challenged sea cucumber fed with the basal diet was significantly higher than those fed with dietary β-glucan or glycyrrhizin. There was no significant difference in cumulative morbidity between the dietary β-glucan and glycyrrhizin treatments over time.     

Impact of several harmful algal bloom (HAB) causing species,on life history characteristics of rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> Müller

In recent years, harmful algal blooms (HABs) have occurred frequently along the coast of China, and have been exhibiting succession from diatom- to dinoflagellate-dominated blooms. To examine the effects of different diatom and dinoflagellate HABs, the life history parameters of rotifers (Brachionus plicatilis Müller) were measured after exposure to different concentrations of HAB species. The HAB species examined included a diatom (Skeletonema costatum) and four dinoflagellates (Prorocentrum donghaiense, Alexandrium catenella, Prorocentrum lima and Karlodinium veneficum). Compared with the control treatment (CT), the diatom S. costatum showed no adverse impacts on rotifers. Exposure to dinoflagellates at densities equivalent to those measured in the field resulted in a reduction in all the life history parameters measured. This included a reduction in: lifetime egg production (CT: 20.34 eggs/ind.) reduced to 10.11, 3.22, 4.17, 7.16 eggs/ind., life span (CT: 394.53 h) reduced to 261.11, 162.90, 203.67, 196 h, net reproductive rate (CT: 19.51/ind.) reduced to 3.01, 1.26, 3.53, 5.96/ind., finite rate of increase (CT: 1.47/d) reduced to 1.16, 1.03, 1.33, 1.38/d, and intrinsic rate of population increase (CT: 0.39/d) reduced to 0.15, 0.03, 0.28, 0.32/d, for the dinoflagellates P. donghaiense, A. catenella, P. lima and K. veneficum, respectively. The results showed that the diatom S. costatum had no detrimental consequences on the reproduction and growth of B. plicatilis, however, the four dinoflagellates tested did show adverse effects. This suggests that dinoflagellate HABs may suppress microzooplankton, resulting in an increase in algal numbers.     

Spatial distribution and dynamics of carbon storage in natural <Emphasis Type="Italic">Larix gmelinii</Emphasis> forest in Daxing’anling mountains of Inner Mongolia,northeastern China

In order to clarify the geographic distribution and change of natural Larix gmelinii forest in carbon storage in Daxing’anling mountains (Great Khingan Range) in North China’s Inner Mongolia, an area-irrelevant conversion equation of biomass from timber volume in the form of B = 0.6966 V was developed by using survey data. Based on the equation, the carbon storage was estimated at 190.172 Tg, and the average carbon density was 41.659 Mg/hm² (area 4.565 × 10⁶ hm²). Carbon stored in standing trees was predominantly distributed in mid-age and mature forests and mainly stocked in the northern part of the study area. Assuming the carbon density of intact area as the maximum value, the potential carbon storage in the entire study area would be 263.674 Tg, approximately 1.4 times of the actual level. Over the period of 1995 to 2010, the carbon storage and carbon density increased by 3.260 Tg and 0.224 Mg/hm², respectively, indicating a weak carbon sink. Comparing with China’s national average level, the carbon density in this area is not as high as expected. Forest quality in terms of carbon capacity is expected to be enhanced by appropriate management schemes under the in-implementation program of forest protection.     

Climate–growth relationship stability of <Emphasis Type="Italic">Picea crassifolia</Emphasis> on an elevation gradient,Qilian Mountain,Northwest China

Climate affects Picea crassifolia growth and climate change will lead to changes in the climate–growth relationship (i.e., the “divergence” phenomenon). However, standardization methods can also change the understanding of such a relationship. We tested the stability of this relationship by considering several variables: 1) two periods (1952–1980 and 1981–2009), 2) three elevations (2700, 3000, and 3300 m), and 3) chronologies detrended using cubic splines with two different flexibilities. With increasing elevation, the climatic factor limiting the radial growth of Picea crassifolia shifted from precipitation to temperature. At the elevation of 2700 m, the relationship between radial growth and mean temperature of the previous December changed so that the more flexible spline had a greater precipitation signal. At the elevation of 3000 m, positive correlation of radial growth with mean temperature and precipitation in September of the previous year became more significant. At the elevation of 3300 m, positive correlation between radial growth and precipitation of the current summer and the previous spring and autumn was no longer significant, whereas the positive correlation between radial growth and temperature of the current spring and summer strengthened. The detrending with the most flexible spline enhanced the precipitation signal at 2700 m, while that with the least flexible spline enhanced the temperature signal at 3300 m. All results indicated that the divergence phenomenon was affected by the climatic signals in the chronologies and that it was most dependent on the detrending method. This suggests it is necessary to select a suitable spline bootstrap for studies of growth divergence phenomena.     

Variation in the emission rate of sounds in a captive group of false killer whales <Emphasis Type="Italic">Pseudorca crassidens</Emphasis> during feedings: possible food anticipatory vocal activity?

This study examines whether a group of captive false killer whales (Pseudorca crassidens ) showed variations in the vocal rate around feeding times. The high level of motivation to express appetitive behaviors in captive animals may lead them to respond with changes of the behavioral activities during the time prior to food deliveries which are referred to as food anticipatory activity. False killer whales at Qingdao Polar Ocean World (Qingdao, China) showed significant variations of the rates of both the total sounds and sound classes (whistles, clicks, and burst pulses) around feedings. Precisely, from the Transition interval that recorded the lowest vocalization rate (3.40 s/m/d), the whales increased their acoustic emissions upon trainers’ arrival (13.08 s/m/d). The high rate was maintained or intensified throughout the food delivery (25.12 s/m/d), and then reduced immediately after the animals were fed (9.91 s/m/d). These changes in the false killer whales sound production rates around feeding times supports the hypothesis of the presence of a food anticipatory vocal activity. Although sound rates may not give detailed information regarding referential aspects of the animal communication it might still shed light about the arousal levels of the individuals during different social or environmental conditions. Further experiments should be performed to assess if variations of the time of feeding routines may affect the vocal activity of cetaceans in captivity as well as their welfare.     

Compensatory growth in hybrid tilapia (<Emphasis Type="Italic">Oreochromis mossambicus ×O. niloticus</Emphasis>) reared in seawater,following restricted feeding

Hybrid tilapia weighing 7.71 g were reared in seawater at 24.0–29.0°C for 8 weeks. The controls were fed to satiation twice a day throughout the experiment, whereas treatment groups were fed at 0.5%, 1.5% or 3.0% body weight per day for 4 weeks, and then to satiation for the remainder of the experiment. During the first 4-week period, there was a curvilinear relationship between growth rate and ration size. Fish fed 0.5% and 1.5% rations displayed compensatory growth response of 2 weeks duration during realimentation. The weight-adjusted growth rate of fish fed at 3% ration was not significantly different from that of the controls by the end of the experiment, when none of the treatment groups had caught up in body weight with the controls. Hyperphagia was observed for the first 2 weeks of realimenatation in fish previously fed at 3% ration, but persisted for the whole realimentation period in groups previously fed at 0.5% and 1.5% rations. None of the feed restricted groups showed improved digestibility, feed efficiency, or protein and energy retention efficiency. Supported by the State Key Laboratory of Freshwater Ecology and Biotechnology of China and Shanghai Fisheries University (Grant No. 200015).     

Production of γ-linolenic acid and stearidonic acid by <Emphasis Type="Italic">Synechococcus</Emphasis> sp. PCC7002 containing cyanobacterial fatty acid desaturase genes

Genetic modification is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. Synechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid (GLA) and stearidonic acid (SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6D, Syd15D and Syd6Dd15D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in Synechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.     

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 of angiotensin-І converting enzyme inhibiting peptide from <Emphasis Type="Italic">Venerupis philippinarum</Emphasis> with nano-liquid chromatography in combination with orbitrap mass spectrum detection and molecular docking

The complexity and diversity of peptide mixture from protein hydrolysates make their characterization difficult. In this study, a method combining nano LC-MS/MS with molecular docking was applied to identifying and characterizing a peptide with angiotensin-? converting enzyme (ACE-I) inhibiting activity from Venerupis philippinarum hydrolysate. Firstly, ethanol supernatant of V. philippinarum hydrolysate was separated into active fractions with chromatographic methods such as ion-exchange chromatography and high performance liquid chromatography in combination. Then seven peptides from active fraction were identified according to the searching result of the MS/MS spectra against protein databases. Peptides were synthesized and subjected to ACE-I-inhibition assay. The peptide NTLTLIDTGIGMTK showed the highest potency with an IC50 of 5.75 μmol L^?1. The molecular docking analysis showed that the ACE-I inhibiting peptide NTLTLIDTGIGMTK bond with residues Glu123, Glu403, Arg522, Glu376, Gln281 and Asn285 of ACE-I. Therefore, active peptides could be identified with the present method rather than the traditional purification and identification strategies. It may also be feasible to identify other food-derived peptides which target other enzymes and receptors with the method developed in this study.     

Isolation of prawn (<Emphasis Type="Italic">Exopalaemon carinicauda</Emphasis>) lipopolysaccharide and β-1, 3-glucan binding protein gene and its expression in responding to bacterial and viral infections

The pattern recognition proteins (PRPs) play a major role in immune response of crustacean to resist pathogens. In the present study, as one of PRPs, lipopolysaccharide and β-1, 3-glucan binding protein (LGBP) gene in the ridge tail white prawn (Exopalaemon carinicauda) (EcLGBP) was isolated. The full-length cDNA of EcLGBP was 1338 bp, encoding a polypeptide of 366 amino acid residules. The deduced amino acid sequence of EcLGBP shared high similarities with LGBP and BGBP from other crustaceans. Some conservative domains were predicted in EcLGBP sequence. EcLGBP constitutively expressed in most tissues at different levels, and the highest expression was observed in hepatopancreas. With infection time, the cumulative mortality increased gradually followed by the proliferation of Vibrio parahaemolyticus and white spot syndrome virus (WSSV). The expression of EcLGBP in response to V. parahaemolyticus infection was up-regulated in hemocytes and hepatopancreas, and the up-regulation in hepatopancreas was earlier than that in hemocytes. EcLGBP expression after WSSV infection increased at 3 h, then significantly decreased in both hemocytes and hepatopancreas. The results indicated that EcLGBP was involved in the immune defense against bacterial and viral infections.