坛紫菜叶状体的抗生素联合除菌效果

为研究坛紫菜及其外生菌群对氯霉素、竹桃霉素和头孢噻肟3种抗生素的敏感性,通过将其与氨苄青霉素、卡那霉素和庆大霉素的组合,优化坛紫菜叶状体的除菌方法,并利用涂布平板、qRT-PCR及16S rRNA测序分析其除菌效果。结果显示,当坛紫菜先以氨苄青霉素(300 mg/L)、卡那霉素(100 mg/L)和庆大霉素(100 mg/L)混合处理18 h,再以氯霉素(50 mg/L)、头孢噻肟(200 mg/L)和竹桃霉素(50 mg/L)混合处理4 d,叶状体的健康率保持在96.3%以上,而对可培养细菌的抑制率达到99.9%。根据qRT-PCR和16S rRNA的结果可知,该种组合方法较前3种抗生素的组合,其菌总数下降41.5%,微生物的丰度和多样性指数明显下降。其中,假单胞菌、交替赤杆菌、交替单胞菌以及海杆菌等得到针对性的抑制。研究表明,多种抗生素的优化组合能够对坛紫菜叶片的菌群结构产生显著影响,使其受到抑制,但仍无法达到绝对除菌的效果。     

‘新海16’愈伤组织诱导及白霜的优化

为获得高品质、活力旺盛且具有分化能力的愈伤组织,本研究在已建立的海岛棉(Gossypium barbadense)‘新海16’再生体系基础上,以MSB附加0.1 mg/L 2,4-D及0.1 mg/L KT为基本培养基,采用单因素随机试验研究外植体不同部位、光照及培养基成分等对‘新海16’愈伤组织诱导的影响。结果表明:‘新海16’愈伤诱导的外植体选用下胚轴中上部绿色茎段诱导效果较优;除此之外,愈伤诱导效果最佳的处理为2 000 lx光照强度、90%MSB无机盐成分、3 g/L PVP及1.2~1.4 g/L Gelrite或Phytagel,均能够降低愈伤组织出现白霜现象,对愈伤组织疏松生长有促进作用。本研究通过优化愈伤组织的诱导方法,提高胚性愈伤组织发生率,为‘新海16’高频再生体系提供科学依据。     

球等鞭金藻三级培养过程中细菌群落多样性分析

球等鞭金藻三级培养过程中生长差异明显,在完全灭菌的一级培养中生长最佳,在未灭菌的三级培养中生长最差。为揭示培养过程中藻际细菌群落多样性与其生长差异的相关性,以球等鞭金藻(Isochrysis galbana)为研究对象,利用Illumina HiSeq平台通过高通量测序的方法对球等鞭金藻三级培养过程中细菌群落多样性进行研究。结果表明,一、二、三级培养组之间的藻际细菌群落组成差异明显。Shannon和Simpson指数分析说明,一级培养组中细菌群落多样性显著低于二级和三级培养组。MetaStat分析发现,一级和三级培养组之间存在两株丰度显著差异的细菌,其中麦氏交替单胞菌(Alteromonas macleodii)在三级培养下丰度显著高于一级,而红球菌(Rhodococcus erythropolis)相反。进一步通过2216E平板涂布法分离并鉴定了麦氏交替单胞菌等17株球等鞭金藻藻际环境细菌;系统进化树构建结果显示,整个进化树分成13个分支,分别对应13个属,包括交替单胞属(Alteromonas)、假交替单胞属(Pseudoalteromonas)、海杆菌属(Marinobacter)等。本研究结果为探索藻菌互作机理奠定了基础,有助于提高水产养殖过程中球等鞭金藻的产量和质量。     

Bacterial communities associated with natural and commercially grown rooibos (Aspalathus linearis)

Aspalathus linearis is a commercially important plant species endemic to the Cape Floristic Region of South Africa and is used to produce a herbal tea known as rooibos tea. Symbiotic interactions between A. linearis and soil bacteria play an important role in the survival of Aspalathus plants in the highly nutrient-poor, acidic fynbos soil. The aim of this study was to characterize and compare rhizosphere and bulk soil bacterial communities associated with natural and commercially grown A. linearis, as well as the effect of seasonal changes on these communities. Bacterial communities were characterized using high throughput amplicon sequencing, and their correlations with soil chemical properties were investigated. The N-fixing bacterial community was characterized using terminal restriction fragment length polymorphism and real time quantitative polymerase chain reaction. Actinobacteria, Proteobacteria, and Acidobacteria were the most dominant bacterial phyla detected in this study. Highly similar bacterial communities were associated with natural and commercially grown plants. Significant differences in the bacterial community were observed between rhizosphere and bulk soils collected in the dry season, while no significant differences were detected in the wet season. This study provides insights into bacterial community structure and potential factors shaping bacterial community structure with commercially important A. linearis.     

甘露寡糖添加方式对哺乳期犊牛瘤胃细菌菌群结构的影响

为利用16S rDNA技术研究甘露寡糖不同添加方式对哺乳期犊牛瘤胃细菌菌群结构影响,本研究选用出生日龄一致、体重接近、健康状况良好的荷斯坦公犊牛20头,随机分为4组。CR组为对照组,饮用乳及开食料中均不添加甘露寡糖;ORa组在饮用乳中添加5 g甘露寡糖,开食料中不添加甘露寡糖;ORb组在开食料中添加5 g甘露寡糖,饮用乳中不添加甘露寡糖;ORc组在饮用乳及开食料中各添加2.5 g甘露寡糖(混合添加)。应用16S rDNA测序技术,研究了甘露寡糖添加方式对犊牛瘤胃细菌构建的影响。结果表明:甘露寡糖添加方式会影响哺乳期犊牛瘤胃细菌总数,但对哺乳期犊牛瘤胃细菌菌群多样性未产生显著影响(P>0.05)。在门水平上,与对照组相比,ORb组颗粒料中添加甘露寡糖显著降低了厚壁菌门与放线菌门的丰度(P<0.05),极显著提高了变形菌门的丰度(P<0.01),优势菌门增加为3种,分别为拟杆菌门、厚壁菌门与变形菌门。在属水平上,甘露寡糖不同添加方式导致哺乳期犊牛瘤胃细菌菌群结发生了改变,其中甘露寡糖不同添加方式对小杆菌属和脱硫弧菌属在瘤胃内丰度均产生了影响,但对ORa组与ORc组犊牛瘤胃细菌影响不及ORb组;且ORb组犊牛瘤胃细菌功能性菌属普雷沃氏-7属、氨基酸球菌属、优杆菌属和琥珀酸弧菌等均发生显著变化。基于以上结果,得出如下结论:本研究条件下,甘露寡糖添加方式对哺乳期犊牛瘤胃细菌菌群多样性未产生显著影响(P>0.05),但开食料中寡糖添加方式对菌群结构产生部分影响,其中发挥瘤胃蛋白降解作用和淀粉降解作用的琥珀酸弧菌科-UCG-001属、利用乳酸的新月形单胞菌属丰度极显著提高(P<0.01),而利用乳糖的优杆菌属丰度显著降低,降解半纤维素的小杆菌属丰度极显著降低(P<0.01)。     

微生物菌群培养组学在动物医学中的应用

实验室条件下可培养的微生物约占自然界中微生物总数的1%，这限制了人们对99%未知微生物的认识和利用，而研究表明，那些“不可培养的微生物”是可以被开发和利用的，未能被纯培养的微生物才是未知微生物的主体。微生物培养组学探索利用多种培养条件和长时间的培养，结合基质辅助激光解吸电离飞行时间质谱法（MALDI-TOF-MS）和16S核糖体RNA（rRNA）测序可以大规模鉴定各种微生物，同时利用全基因组测序和宏基因组测序手段对未知微生物进行深入分析。本文综述了国内外近年来微生物菌群培养组学在反刍动物胃肠道、禽类盲肠及家畜鼻腔微生物菌群研究中的最新进展，探讨将动物体内菌群培养组学方法应用于动物疾病防治领域的可行性。作为一个新兴的研究方法，尽管该培养组学还存在一些不够成熟的方面，但它的发展前景十分广阔，微生物菌群培养组学方法和其他研究方法的互补已经逐渐成为发展兽医微生物学新的突破口。     

Formulated diet alters gut microbiota compositions in marine fish Nibea coibor and Nibea diacanthus

Nibea, an economical marine fish, is generally fed on trash fish (the low‐value fish), which can cause high feed costs and waste pollutions in high‐density aquaculture. To assess the effect of formulated diet on the gut microbiota in Nibea coibor and Nibea diacanthus, denaturing gradient gel electrophoresis, clone libraries analysis and Illumina sequencing of the 16S ribosomal RNA gene were used in this study. Two Nibea fishes were both dominated by Gammaproteobacteria (especially Photobacterium) and shared a set of gut microbiota, including Bacilli, Mollicutes, Alphaproteobacteria and Fusobacteriia. Statistical analyses revealed that formulated diet led to lower feed conversion ratio (p < 0.001), lower abundance of Vibrio (p = 0.040) and infectious diseases pathways (p = 0.001), higher abundance of polysaccharide‐degrading bacterium Cellvibrio (p = 0.006) in two Nibea species, with higher weight gain rate (p = 0.023) and microbial diversity (Shannon, p = 0.049 and Simpson, p = 0.044) and more carbohydrate metabolism (p = 0.020) observed in N. coibor. The distribution and correlation network of 17 potential short‐chain fatty acid producing bacteria were obtained and visualized in all treatment groups. The results reveal that formulated diet has beneficial effects on the gut microbial ecology in two Nibea fishes, which suggests the possibility of replacing trash fish diet with formulated diet in Nibea aquaculture.     

Using genetic methods for analysis of fish meals and feeds employed in Greek mariculture

Large quantities of high protein fish meals are needed to sustain cultured species and thus the impact to marine ecosystem has been highly discussed. The aim of this study was to apply a PCR‐cloning methodology for a robust insight into the composition of commercial fish meals and feeds for farmed species of the Greek mariculture, assessing the risk posed by aquaculture to marine ecosystems but also the risk posed by commercial fish feeds to the increase in trophic level of species farmed in Greece. 89% of the sequences were identified to species level and only 11% to genus/family level. Overall, a total of 49 taxa were identified (44 fish species/taxon, five non‐fish species/taxon). Even though small pelagic fish like Engraulis sp. were the main portion, a wide range of species constituted the fish meals and feeds. Plant and animal species were also detected as an alternative protein source. Feed products employed in Greek mariculture still contain large portions of fish meals which increase the mean trophic level of farmed species causing a farming up trend. The results emphasize that such molecular methodologies are needed to certify aquafeeds allowing fish feed producers to demonstrate their commitment to sustainable aquaculture.     

辣椒新品种天椒16 号的选育

天椒16 号是以45-2-1-1-1-1 为母本，以76-1-1-1-1-1 为父本配制而成的辣椒一代杂种。中熟，始花节位为第 10～12 节。果实羊角形，青熟果深绿色，果面皱，果长25.56 cm，果径5.83 cm，单果质量103.8 g，辣味淡，具有良好的商 品性。每667 m² 产量可达5 000 kg 以上，田间对辣椒病毒病、疫病、炭疽病和白粉病的抗性强于对照陇椒2 号。适宜在甘 肃省进行保护地和露地栽培。