甘露寡糖对泥鳅生产性能、肠道菌群和非特异性免疫功能的影响

【目的】探讨甘露寡糖对泥鳅生产性能、肠道菌群和非特异性免疫功能的影响。【方法】选择健康均重为(3.27±0.05)g泥鳅360尾,随机分为6组,每组3个重复,每个重复20尾。在基础饲料中分别添加0(对照组)、100、200、400、600和800 mg/kg甘露寡糖。试验期70 d。检测成活率、特定生长率、肥满度、饵料系数、肠体指数、肠长指数,肠道大肠埃希菌、乳酸杆菌、双歧杆菌以及血浆和肝胰脏溶菌酶、超氧化物歧化酶、过氧化氢酶等指标。【结果】①200和400 mg/kg组特定生长率、肥满度、肠体指数和肠长指数显著高于对照组,饵料系数显著低于对照组(P<0.05);②200和400 mg/kg组肠道乳酸杆菌和双歧杆菌数量显著高于对照组,大肠埃希菌数量显著低于对照组(P<0.05);③200和400 mg/kg组血浆和肝胰脏SOD、CAT活性显著高于对照组(P<0.05),血浆LZM含量显著高于对照组(P<0.05)。【结论】在饲料中添加200～400 mg/kg甘露寡糖可促进泥鳅肠道发育,优化肠道微生态环境,显著提高其非特异性免疫功能和生长性能。

有机和常规农业管理方式下的浮游植物群落特征及其影响因素

为了解不同农业施肥管理方式对水体浮游植物群落的影响,本研究以江苏句容戴庄有机示范村和常规农业管理区池塘浮游植物为对象,分析冬夏两季浮游植物群落特征及相关水环境因子.结果表明:冬季有机和常规农业区域的浮游植物分别检到7门95种和7门111种,夏季分别检到7门102种和6门112种,有机农业区浮游植物由冬季的隐藻—绿藻型向夏季的蓝藻—绿藻型变化,常规农业区从隐藻—绿藻型向蓝藻—硅藻型变化,蓝藻逐渐取代隐藻的优势地位.夏季各池塘浮游植物alpha多样性没有显著的区域差异,但冬季常规农业区浮游植物物种丰富度、蓝藻和裸藻的Shannon-Wiener多样性指数显著较高.指示物种分析显示,谷皮菱形藻(Nitzschia palea)和梅尼小环藻(Cyclotella meneghiniana)为有机农业区指示物种,而梭形裸藻(Euglena acus)、针形纤维藻(Ankistrodesmus acicularis)、肘状针杆藻(Synedra ulna)和狭形纤维藻(Ankistrodesmus angustus)为常规农业区指示物种,此6种藻为中、富营养化藻类,说明水体氮磷营养盐浓度较高,这与周围农田氮磷流失密切相关.Pearson相关性分析表明,有机农业区浮游植物alpha多样性与氮磷浓度的相关性不显著,但常规农业区浮游植物物种丰富度和Pielou均匀度与磷酸盐、总氮、氨氮和pH值显著相关.Mantel检验表明,有机农业区群落相异性仅在冬季受到总氮和氨氮的显著影响;而常规农业区在冬夏季受到氮磷两类营养盐的交替影响.本研究结果初步揭示了有机和常规农业区水体浮游植物群落结构特征及其影响因素,为区域水生生物多样性保护与稻田施肥管理的优化提供科学基础.

Identification of a marine agarolytic bacterium Agarivorans albus QM38 and cloning and sequencing its beta-agarase genes

A total of 117 agar-decomposing cultures were isolated from coastal seawater around Qingdao, China. The phenotypic and agarolytic features of an agarolytic isolate, QM38, were investigated. The strain was gram negative, strictly aerobic, curved rod and polar flagellum. On the basis of several phenotypic characters, biochemical and morphological characters and phylogenetic analysis of the gene coding for the 16S rRNA, the strain was identified as Agarivorans albus strain QM38. This strain can liquefy the agar on the solid agar plate. An excellular agarase activity was determined in liquid culture. The enzyme exhibited maximal activity at 40 ℃, pH 7.6. Its activity was greatly affected by different concentrations of agarose. The highest activity 32 U/ml was achieved in the culture supernatant. The hydrolytic product was analyzed by fluorophore-assisted carbohydrate electrophoresis （FACE）. After complete hydrolysis of agarose, a series of agaro-oligosaccharides were produced. The main products of the enzymes were oligosaccharides in the degree of polymerization （DP） of 2, 4, 6 and 8. Three genes agaD01, agaD02 and agaD03, encodingβ-agarases, had been cloned from genomic DNA of Agarivorans albus strain QM38. The open reading frame of agaDOl, consisted of 2 988 bp, and shared 95.5%-98.9% identity to the β-agarase genes of some strains of Vibrio and Agarivorans. Gene agaD02 comprised 2 868 bp and encoded a 955- amino-acid protein. It showed 97.4% and 98.7~0 identity to the β-agarase genes of strain Vibrio sp. PO-303 and strain Vibrio sp. JT0107, respectively. Only partial sequence of agaD03 gene has been cloned. It showed 96.5% identity to β-agarase gene （agaB） of Pseudoalteromonas sp. CY24, and shared 96.8% identity to β-agarase-c gene of Vibrio sp. PO-303.

Isolation and diversity analysis of heterotrophic bacteria associated with sea anemones

A study was undertaken to investigate the heterotrophic bacterial flora associated with the sea anemones. Samples of the sea anemones Anthopleura midori were collected from the coast of Weihai and bacteria were isolated from these samples. Additionally, high numbers of viable bacteria were obtained from the celom wall and surface of anemone, the community of cultivable bacteria was very diverse. As a result of this isolation, 60 strains were obtained, 56 of them were selected for identification and characterization by 16S rRNA gene sequence analysis and limited phenotypic testing. Among these isolates, 16 strains were phylogenetically related to members of the genus Pseudoalteromonas and neighboring taxa. Other isolates included members of the genera Colwcllia, Vibrio, Acinetobactcr, Pseudomonas, Endozoicomonas, Roseovarius, Paraeoceus, Loktanella, Leisingcra, Sulfitobacter, Bacillus, Staphylococcus, Plantibacter, Microbaeterium, Micro- coccus, Joostella, Psychroserpens, Cellulophaga, Krokinobaeter, Polaribacter and Psychrobaeter. Seven potential novel species were found. Among 60 strains, 17 of them can produce proteolytic exoenzyme, 20 can produce lipolytic exoenzyme. Strain NQ8 has strong antagonistic effects on some Vibrio strains. This study demonstrates that the culturable fraction of bacteria from the sea anemones Anthopleura midori is diverse and appears to possess much potential as a source for the discovery of novel bioactive materials.

灰褐链霉菌与黄色短杆菌属间原生质体融合提高ε-聚赖氨酸产量（英文）

为提高灰褐链霉菌(Streptomyces griseofuscus LS-6)的ε-聚赖氨酸(ε-poly-lysine,ε-PL)发酵水平,从增加内源性前体赖氨酸角度考虑,将S. griseofuscus LS-6与赖氨酸生产菌株黄色短杆菌(Brebvibacterium flavum S62)进行属间的原生质体融合。以S. griseofuscus LS-6产黑色素作为遗传标记挑选到1株融合子LS-32,其ε-PL摇瓶产量为2.82 g/L,是亲本菌株LS-6的1.73倍;LS-32在补料分批发酵中的ε-PL产量达到56.4 g/L,比亲本提高了38.9%。随机引物扩增多肽DNA技术显示融合子LS-32中确实有来自B. flavum S62的基因;融合子ε-PL代谢途径中关键酶的活力(己糖激酶、天冬氨酸激酶、ε-PL合成酶等)和胞内氨基酸含量(赖氨酸、谷氨酸、精氨酸等)均高于亲本,是LS-32高产ε-PL的重要原因。本研究明确了内源性赖氨酸对于提升ε-PL产量的重要性,且属间融合为ε-PL产生菌的育种研究提供了新的参考途径。

Archaeal diversity and abundance within different layers of summer sea-ice and seawater from Prydz Bay,Antarctica

Fluorescence in-situ hybridization （FISH） and 16S rRNA gene clone library analyses were used to determine the abundance and diversity of archaea in Prydz Bay, Antarctica. Correlation analysis was also performed to assess links between physicochemical parameters and archaeal abundance and diversity within the sea-ice. Samples of sea-ice and seawater were collected during the 26th Chinese National Antarctic Research Expedition. The results of FISH showed that archaea were relatively abundant within the top layer of the sea-ice, and correlation analysis suggested that the concentration of 4NH＋ might be one of the main factors underlying this distribution pattern. However, using 16S rRNA gene libraries, archaea were not detected in the top and middle layers of the sea-ice. All archaeal clones obtained from the bottom layer of the sea-ice were grouped into the Marine Group I Crenarchaeota while the archaeal clones from seawater were assigned to Marine Group I Crenarchaeota, Marine Group II Euryarchaeota, and Marine Group III Euryarchaeota. Overall, the ifndings of this study showed that the diversity of archaea in the sea-ice in Prydz Bay was low.

稻田和池塘养殖模式对中华绒螯蟹肠道菌群、免疫酶活及肌肉游离氨基酸的影响

为了探究稻田和池塘养殖模式对中华绒螯蟹(Eriocheir sinensis)肠道菌群、免疫力和肌肉风味影响,选取同一批次稻田和池塘的中华绒螯蟹在5月和10月对其肠道菌群、免疫酶活和肌肉游离氨基酸进行测定。结果表明:中华绒螯蟹肠道优势菌群为软壁菌门、拟杆菌门、变形菌门和厚壁菌门。在季节和养殖模式均对蟹的肠道菌群造成影响的情况下,季节变化对菌群变化的影响更大。从5月到10月,稻田养殖模式下中华绒螯蟹肠道菌群的丰富度和多样性均出现上升趋势,而池塘养殖模式下肠道菌群的丰富度和多样性则无显著变化。两种养殖模式下中华绒螯蟹肠道中的超氧化物歧化酶(SOD)在5月的活性均高于10月,在10月池塘模式下的免疫酶活性溶菌酶(LZM)和酸性磷酸酶(ACP)显著高于稻田模式。各组肌肉中均检测出14种呈味氨基酸,呈味氨基酸中甜味氨基酸含量(TSAA)>总苦味氨基酸(TBAA)>总鲜味氨基酸(TUAA),且池塘养殖模式下中华绒螯蟹肌肉中TSAA和总游离氨基酸TFAA高于稻田养殖模式下含量。因此,池塘养殖模式较稻田养殖模式下的中华绒螯蟹肠道菌群时间上更稳定,免疫酶活更强,肌肉风味更优。