生物絮团技术在水产养殖中的应用研究

传统的水产养殖模式所带来的环境污染、资源浪费和病害频发等问题已成为制约我国水产养殖业可持续发展的主要因素。生物絮团技术(BFT)具有净化水质、提高饵料利用率及病害防控等优点,被认为是有望解决上述问题的新型健康生态养殖技术,已在国内外得到一定规模的应用,并获得了良好的经济、社会和生态效益。本文重点介绍了生物絮团的形成与培养、生物絮团的主要影响因素及其在水产养殖中的应用效果。研究认为,BFT能够改良水质、节约养殖用水、降低饲料成本、提高养殖对象存活率、增加养殖产量和效益;将BFT与生物膜技术相结合,能够更有效地维持养殖水体中适宜的生物絮团含量,避免生物絮团的过量沉积,并能提高水质改良及增产增收的应用效果,具有广阔的应用前景。     

Analysis of bacterial community and bacterial nutritional enzyme activity associated with the digestive tract of wild Chilean octopus (Octopus mimus Gould, 1852)

Information on the bacterial community associated with octopus is very scarce, unlike fish and other molluscs. This study revealed the bacterial community associated with digestive tract of wild Chilean octopus Octopus mimus using a culture‐dependent method and 16S rDNA clone library. Moreover, we analysed the bacterial nutritional enzyme activity of culturable bacteria. A culture‐dependent method showed that the composition of the culturable bacterial community was substantially different between female and male octopus. The predominant species in female octopus were Vibrionaceae and Streptococcaceae, whereas only Vibrionaceae was dominated in male octopus. Bacterial nutritional enzyme activities of culturable bacteria from male octopus were much higher than female octopus. The 16S rDNA clone library analysis showed that the bacterial community of male octopus exhibited a higher diversity than that of female octopus. The genus Mycoplasma was the predominant bacteria in the digestive tract of all octopus samples. The results obtained in this study raise the possibility that each octopus has different food consumption due to different bacterial community and nutritional enzyme activity, although Mycoplasma sp. is one of the predominant bacteria in the digestive tract. Moreover, our results are useful for the future of microbiological investigation associated with the octopus and for probiotics in the octopus aquaculture.     

不同健康程度和抗生素氟苯尼考干预下斑石鲷肠道菌群的结构差异

为了研究不同健康程度和抗生素氟苯尼考干预下斑石鲷肠道菌群结构的差异及其与养殖环境中菌群结构的相关性,采用Illumina Hi Seq PE250高通量测序的方法对健康、亚健康、典型黑身病和口服氟苯尼考条件下的斑石鲷肠道、养殖水体和颗粒饵料中的细菌多样性及群落结构进行了分析比较。结果显示,养殖水体中细菌多样性高于肠道和颗粒饵料。不同健康程度及氟苯尼考干预下斑石鲷肠道中细菌均以变形菌门、厚壁菌门和软壁菌门为主,且对应的操作分类单元(OTU)占样品全部OTU的比例均达到85%以上。黑身病的发生可影响斑石鲷肠道中丰度最高的前20种优势细菌种类的排名次序,其中变形菌门中的弧菌属的相对丰度显著增加,且随着弧菌属丰度的增加,斑石鲷的黑身病症状也逐渐加重。饵料中添加氟苯尼考投喂斑石鲷能使患病鱼肠道弧菌属的丰度从60.33%下降到1.29%,较大程度改变了肠道的菌群结构,并证实氟苯尼考有效防治黑身病。其次,养殖水体和颗粒饵料对斑石鲷肠道菌群也有一定影响,且养殖水体的影响高于颗粒饵料。本研究首次报道了斑石鲷肠道菌群结构,其研究结果为今后斑石鲷的健康养殖、疾病防控及其微生态学研究提供了参考依据和技术支撑。     

Contribution of microorganisms to the biofilm nutritional quality: protein and lipid contents

The nutritional quality of biofilm, a microbial community associated to an organic matrix, was evaluated in artificial substrate (polyethylene screen) in net cages during 30 days in the Patos Lagoon estuary, Southern Brazil. During this period, samples of biofilm were collected each 5 days for analysis of chlorophyll a, microorganisms abundance, dry weight, protein and lipid contents. During the study, chlorophyll a varied from 0.38 to 2.75 μg cm^?2; dry weight between 7.16 and 17.63 mg cm^?2; protein content from 0.43 to 1.76 mg cm^?2 and lipid concentration between 1.21 and 4.23 mg cm^?2. The variation of lipid in the biofilm was closely related to the abundance of free heterotrophic bacteria (34.25–56.54 × 10⁶ cells cm^?2), filamentous cyanobacteria (7.5–15.9 × 10⁶ filaments cm^?2), flagellates (6.92–12.89 × 10⁶ cells cm^?2) and mainly nematodes (29–1,414 organisms cm^?2), while protein content varied similarly to the abundance of unicellular centric diatoms (52.10–179.81 × 10³ cells cm^?2), and nematodes. This information will allow a better management of food supply to raised aquatic organism with the utilization of natural productivity in the culture systems, with considerable decrease in production costs.     

The influence of salmon surface mucus on the growth of Flavobacterium columnare

Flavobacterium columnare is the causative agent of columnaris disease. The presence of lesions on the gills, skin and fins of diseased fish suggests that F. columnare is able to utilize fish skin mucus as a substrate for growth and that exposure to this material would alter the expression of genes involved in the colonization of the outer surfaces of the fish. Growth, biofilm formation, extracellular protease production and changes in protein expression of F. columnare strain C#2 cultured in media supplemented with juvenile Atlantic salmon skin mucus were compared with the same media without mucus. C#2 was able to grow by using mucus as the sole nutrient source. Growth in mucus-containing media induced cells to grow as a biofilm and extracellular protease activity increased in mucus-containing cultures. SDS-PAGE protein profiles showed that expression of six extracellular proteins increased in mucus-containing media. These results demonstrate that salmon surface mucus promotes the growth of F. columnare and that exposure to mucus alters the growth characteristics of this bacterium with regard to protease production and biofilm formation. Further characterization of mucus-induced physiological changes will increase our understanding of the basis of virulence of this economically important fish pathogen.     

博斯腾湖沉积物氨氧化细菌的筛选与鉴定

为了解干旱区淡水湖泊沉积物微生物在氨氧化中的功能,利用铵盐培养基对博斯腾湖不同深度沉积物进行驯化培养,结合水环境因子分析混合菌株产亚硝酸盐能力,从中分离纯化氨氧化细菌并对其氨转化能力进行试验。结果表明,6个采样位点12个样品中,11号位点深层沉积物(20～40 cm)混合菌株能有效利用铵盐产生亚硝酸盐,且连续6 d稳定维持其质量浓度高达20 mg/L以上。进而,从该混合菌液中分离得到30株单菌,其中有2株呈现出明显的氨氧化细菌特性。通过16S rRNA基因序列比对及形态学分析分别鉴定其为弯曲芽孢杆菌(Bacillus flexus)和赤红球菌(Rhodococcus ruber)。     

水稻革兰氏阳性细菌的主要种群结构及对纹枯病和恶苗病菌的拮抗性

于1998~2004年对采自浙江、江苏、福建和云南省的756份稻株和稻种样本进行了革兰氏阳性细菌的分离、鉴定研究。被分离的1015个菌株经致病性测定、菌落形态及部分细菌学特征（革兰氏染色、KMB培养基上的荧光色素及芽孢的染色镜检等）测定后,选出代表菌株74个连同5个对照菌株用Biolog及脂肪酸分析法(FAME)进行测试, 鉴定出Bacillus属5个种及其他3属的革兰氏阳性细菌,并发现枯草芽孢杆菌和巨大芽孢杆菌菌株具有很好的纹枯病和恶苗病拮抗能力,但来自短小芽孢杆菌和巨大芽孢杆菌的极少数菌株在条件适宜时能与其他病原菌一起引起水稻褐斑,从这些种筛选生防菌株时应充分考虑其风险。     

含白叶枯病抗性基因Xa23水稻恢复系的分子标记辅助选育

 通过常规杂交育种技术和分子标记辅助选择技术,对携有白叶枯病抗性基因Xa23的亲本材料CBB23进行回交转育和农艺性状改良。通过分子标记辅助选择检测目的基因,快速获得了12份具有Xa23基因的恢复系稳定材料。采用白叶枯病菌株的专化强毒菌系P6对该12份材料进行室内接种鉴定,筛选出1份高抗白叶枯病的抗性新恢复系材料。应用3个不同的不育系对该恢复系进行测交,结果表明该恢复系具有良好的恢复性及产量潜力。     

水稻白叶枯病抗性基因鉴定进展及其利用

 1982~1987年，日本热带农业研究中心（TARC）和国际水稻研究所（IRRI）合作采取统一研究方案，创建了国际水稻白叶枯病抗性鉴别系统。统一了命名，删去了重复（包括与Xa3相同的Xa4b、Xa6和xa9）。截至2005年6月，经国际注册确认和期刊报道的水稻白叶枯病抗性基因共30个，其中 Xa22(t)、Xa26(t)、xa26(t)、Xa27(t)、xa28(t)、Xa29(t)和3个Xa25(t)为暂定名基因，有待订正。在30个基因中，21个为显性基因（Xa），9个为隐性基因（xa）；13个表现全生育期抗性，15个为成株期抗性，Xa21和Xa25(t) (O. minuta)两个基因在分蘖后期表达抗性。已被定位的抗性基因有17个，即第11染色体上有7个，Xa3、Xa4、Xa10、Xa21、Xa22(t)、Xa23和Xa26；第4染色体上有4个，Xa1、Xa2、Xa12和Xa14;第5染色体上有2个，xa5和xa13；第6染色体上有Xa7和Xa27；第12染色上有Xa25(t)；第1染色体上有Xa29（t）。包括Xa1、xa5、Xa21、Xa26五个基因已被克隆。并讨论了合理利用抗性基因等问题。