凡纳滨对虾养殖体系中群体感应淬灭菌的筛选、安全性评估及发酵条件优化

从凡纳滨对虾(Litopenaeus vannamai)养殖体系中筛选得到具有群体感应淬灭(Quorum quenching, QQ)活性的潜在功能菌，并对菌株进行16S rDNA鉴定、安全性评估及发酵条件优化。采用根癌农杆菌(Agrobacterium tumefaciens) A136液体X-gal法进行活性菌株筛选，发现2株细菌(BDZ5和W1B)的发酵液对信号分子己酰基高丝氨酸内酯(C6-HSL)具有显著的降解作用，但经沸水浴处理后均丧失降解活性。经16S rDNA基因序列分析，菌株BDZ5和W1B分别被鉴定为芽孢杆菌属(Bacillus)和科贝特氏菌属(Cobetia)。安全性评估结果显示，菌株BDZ5和W1B均无溶血性，且对多数抗生素具有敏感性，注射2株活性菌未提高凡纳滨对虾死亡率，且对凡纳滨对虾血清免疫酶活性无显著影响。单因素条件优化结果显示，在起始pH为7.0、盐度为20、0.0005%的MnCl2条件下培养48 h后，菌株BDZ5达到最大生长量；在起始pH为6.0、盐度为40、0.05%的CaCl2条件下培养48 h，菌株W1B达到最大生长量。2株菌的最适信号分子C6-HSL降解条件与其生长条件有所不同，在起始pH为7.5、盐度为30、0.0005%的MnCl2条件下培养48 h，2株活性菌对信号分子C6-HSL的降解率达到最大值。研究表明，菌株BDZ5和W1B可作为益生菌应用于水产养殖，为基于QQ角度防治对虾细菌性病害提供优良菌种。     

增氧方式对水芹菜-微生物联合作用处理冬季养殖废水的影响

为探究不同增氧方式对水芹菜-微生物联合作用处理养殖废水消化液及微生物群落结构变化的影响，设置循环和曝气两组试验，以静置处理作为对照，共运行35 d，间隔7 d取样，测定常规水质指标，在试验开始和结束时分别测定水芹菜生长情况，并采用高通量测序技术对微生物群落结构进行分析。结果显示，经过35 d的处理，静置组、曝气组和循环组的DO（溶解氧）分别达到0.60、10.38 mg·L^-1和10.85 mg·L^-1；静置培养的水芹菜长势最好，相对生长速率为0.03 g·g^-1·d^-1；三种处理方式对水质的净化效果差异显著，循环组的水质净化效果最好，COD（化学需氧量）、NH₃-N（氨氮）和TN（总氮）的去除率分别达89.89%、69.40%和77.65%；黄杆菌属（Flavobacterium）的相对丰度从43.21%下降到0.16%，克里斯滕森菌属（Christensenellaceae）和瘤胃球菌属（Ruminococcaceae）的相对丰度则大幅增加。水芹菜在冬季于养殖废水消化液中可以正常生长，曝气和循环处理可以明显提高净化效率；黄杆菌属和假单胞菌属（Pseudononas）是脱氮的功能菌属，梭菌纲（Clostridia）的大量繁殖有利于COD的去除。     

皖北石灰岩山地不同植被恢复模式对植物群落物种组成及多样性的影响

通过研究皖北石灰岩山地生态系统的植被群落,分析不同植被恢复群落的组成、结构、多样性和生活型组成,进而阐明石灰岩山地生态系统植被恢复模式对群落结构特征以及植物物种多样性的影响。结果表明:研究区有54种植物,隶属26科54属。侧柏(Platycladus orientalis)+构树(Broussonetia papyrifera)人工混交林有植物种类16科26种,分别占总数的61.5%和48.1%。酸枣(Ziziphus jujuba var.spinosa)+牡荆(Vitex negundo var.cannabifolia)灌丛和荩草(Arthraxon hispidus)+牡荆草灌丛植物种类分别为32和33种,分别占调查总物种数的59.3%和61.1%。不同植被恢复群落生活型也有差异。侧柏+构树人工混交林群落高位芽植物占优势,而酸枣+牡荆灌丛和荩草+牡荆草灌丛则以地面芽植物所占比例较大。分析表明,多样性指数、均匀度指数和优势度指数均表现为侧柏+构树混交林>酸枣+牡荆灌丛>荩草+牡荆草灌丛,但物种丰富度表现为荩草+牡荆草灌丛>酸枣+牡荆灌丛>侧柏+构树混交林。Sörensen较高值出现在酸枣+牡荆灌丛与侧柏+构树混交林和荩草+牡荆草灌丛之间,侧柏+构树混交林与荩草+牡荆草灌丛Sörensen多样性值较低。研究结果表明,石灰岩山地独特的地貌特征和人工植被恢复对植物群落组成、多样性及其生活型组成产生重要影响。     

Resveratrol protects cortical neurons in infant rats with bacterial meningitis through miR-223-3p/NLRP3 pathway

AIM To investigate the protective effect of resveratrol (Res) on cortical neurons in rat bacterial meningitis (BM) model. METHODS Group B hemolytic Streptococcus was injected via the posterior cistern to establish a BM model. Resveratrol was administered intranasally and microRNA-223-3p (miR-223-3p) antagomir was administered by intracerebroventricular injection. HE staining was used to observe the pathological changes of the brain tissue. Loeffler scoring method was used to evaluate the neurobehavioral functions. TUNEL staining was used to detect neuronal apoptosis. The expression of interleukin-1β (IL-1β), IL-18, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Iba1) was detected by immunofluorescence staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）, cleaved caspase-1, IL-1β and IL-18 were determined by Western blot. The expression level of miR-223-3p was detected by RT-qPCR. Online software TargetScan was used to search for the complementary nucleotide sequences between miR-223-3p and NLRP3 mRNA. RESULTS Compared with sham group, the thickness of meninges in BM model was increased, the neurological score was decreased (P<0.05), and the number of TUNEL positive neurons was increased significantly (P<0.05). Astrocytes and microglia were activated, the fluorescence intensity of IL-1β and IL-18 was increased (P<0.05), and the expression levels of NLRP3, cleaved caspase-1, IL-1β, IL-18 and miR-223-3p were increased (P<0.05). Compared with BM group, after treatment with resveratrol, the neurological score was increased (P<0.05), the number of TUNEL positive neurons was decreased significantly (P<0.05), and the inflammatory response of astrocytes and microglia was suppressed. The fluorescence intensity of IL-1β and IL-18 was decreased (P<0.05), the protein levels of NLRP3, cleaved caspase-1, IL-1β and IL-18 were decreased (P<0.05), and the expression level of miR-223-3p was increased (P<0.05). A nucleotide sequence in the 3'-UTR of NLRP3 mRNA might be targeted by miR-223-3p. In the brain of rat BM model, compared with antagomir control group, the expression of NLRP3 was increased in miR-223-3p antagomir group with resveratrol treatment (P<0.05). CONCLUSION Resveratrol may reduce the inflammatory death of cortical neurons in BM model of infant rats through miR-223-3p/NLRP3 pathway, thus playing a protective role for the neurons.     

温度对拟穴青蟹循环水养殖系统微生物群落结构的影响

为探究温度对拟穴青蟹(Scylla paramamosain)循环养殖水体中微生物群落的影响,深入理解环境因子与养殖水体微生物群落之间的相关性。在水温分别为18℃、22℃、26℃、30℃和32℃的循环养殖系统中进行拟穴青蟹养殖,采用16S rRNA高通量测序技术对不同温度下的养殖水体微生物群落组成进行分析。结果表明:(1) Alpha多样性指数显示5组水样中细菌群落多样性指数大小为t26t22t30t32t18,物种丰度大小为t22t26t32t30t18。温度可以导致养殖水体中微生物群落多样性发生变化,但并未发现二者之间直接的相关性。(2) 5种温度下的养殖系统中细菌群落共有31门、43纲、94目、185科和351属,变形菌门(Proteobacteria)为循环养殖水体中绝对优势菌门。在养殖系统水温为26℃时,养殖水体中氮磷污染物含量相对较高,其水中脱氮除磷菌属的相对丰度也显著高于其他4组。(3)由花瓣图和UPGMA聚类分析图进行相似性分析可知,水体微生物群落对温度的变化有着良好的响应, 5组水样的细菌群落结构总体相似性较低,其中最高温度32℃和最低温度18℃的水体中细菌群落结构最为相似。(4)水体中微生物群落结构和与环境因子的相关性分析显示,氮磷营养盐中的总磷(TP)和硝酸盐(NO3–-N)对水中微生物群落特征具有显著性影响,而温度是导致养殖水体中氮磷营养盐差异化的主要原因。上述研究结果初步解释了温度对拟穴青蟹循环养殖水微生物群落的作用机制,为下一步深入研究拟穴青蟹循环水养殖系统中环境变化与微生物的相互关系提供参考。     

斑点叉尾鮰养殖池塘底泥微生物群落结构特征及其影响因素

微生物群落在水产养殖环境中起着重要作用，深入了解微生物群落组成及其构建机制对了解池塘生态功能具有重要意义。本研究通过高通量测序技术研究斑点叉尾鮰（）养殖池塘底泥中细菌群落机构特征，并分析了细菌群落的主要影响因素。结果显示，斑点叉尾鮰养殖池塘底泥细菌群落结构呈现出季节性变化，冬春季样品相似性较高。斑点叉尾鮰养殖池塘底泥优势菌群主要为变形菌门（Proteobacteria）、绿弯菌门（Chloroflexi）、拟杆菌门（Bacteroidetes）、厚壁菌门（Firmicutes）和酸杆菌门（Acidobacteria），与池塘水体细菌群落间有明显的差别。线性判别分析显示，冬、春和秋季的特异性优势菌群较为丰富，冬季和春季分布较为集中，分别集中于浮霉菌、拟杆菌门、绿弯菌门和厚壁菌门、拟杆菌门，秋季特异性优势菌群分布特别分散，而夏季优势细菌类群最少，只有绿弯菌门厌氧绳菌属1个属具有显著优势。环境因子中，透明度和总溶解性悬浮物与斑点叉尾鮰养殖池塘底泥细菌群落结构有显著相关（<0.05），对细菌群落结构的影响也最大。本研究旨为斑点叉尾鮰池塘微生物群落的变化和调控提供借鉴。     

浅谈大兴区社区蔬菜技术服务

为提升社区居民的生活满意度,进一步探索阳台蔬菜管理技术、种植品种和管理模式,大兴区蔬菜技术推广站开展了基层调研、专业方案设计、观摩培训等社区蔬菜技术服务工作,并取得了较好的效果:服务范围不断扩大、栽培设备及配套技术逐渐完善、社区氛围良好等;并对发展前景进行了展望:未来将会向省空间、智能化、低碳化的多功能蔬菜种植模式发展。     

水分胁迫对典型草原植物群体光合作用的影响

基于锡林浩特国家气候观象台2016—2017年的群体光合作用及同期土壤水分观测数据,分析了不同水分胁迫状态下典型草原植物群落光合作用的受限程度。结果表明:牧草生长初期及末期,群体光合速率处于较低水平,水分胁迫未对群体光合作用产生明显影响,7月牧草进入旺盛生长期,群体光合速率显著升高,并在8月上旬达到最高值(≥10.89μmol·m~(-2)·s~(-1)),水分限制对群体光合作用的影响显著;在牧草生长的初期和中期,群体光合速率、呼吸速率与10～20 cm土层土壤水分含量相关性最显著,表明植物根系和土壤微生物在该层分布最为集中;0～30 cm各土层平均土壤水分含量与群体光合速率之间存在良好的线性回归关系,方程拟合效果为极显著,表明30 cm以上土层土壤水分含量影响典型草原植物群体光合作用;MK突变检验分析表明,当典型草原0～10 cm土层土壤体积含水量低于9%时(对应重量含水量为6.82%,相对含水量为27.60%)、0～30 cm土层平均土壤体积含水量低于12%时(对应重量含水量为9.57%,相对含水量为39.24%),群体光合速率会受到非常明显的抑制。综合分析,在典型草原干旱预报预警中,可将0～10 cm土层土壤体积含水量低于9%、0～30 cm土层平均土壤体积含水量低于12%视为典型草原发生中度干旱时对应的土壤水分临界值。     

添加芽孢杆菌对草鱼池塘中真核微生物的影响

为了研究添加芽孢杆菌对池塘中真核微生物群落结构和理化因子的影响,采用高通量测序技术分析了实验组(添加芽孢杆菌池塘)与对照组(普通池塘)水体真核微生物群落结构,同时分析了两组池塘的水体理化指标。结果表明:8、9月实验组池塘水体中TN、NH_4~+-N、NO_3~--N含量显著低于对照组(P0.05)。水体中梅尼小环藻(Cyclotella meneghiniana)、红囊藻(Hedriocystis)、蓝隐藻(Chroomonas)、丝孢酵母属(Trichosporon)和小环藻属(Cyclotella)真核微生物丰度显著高于对照组(P0.05)。实验组池塘水体真核微生物Chao1指数和Shannon指数显著高于对照组(P0.05)。实验结果证实:通过向池塘添加芽孢杆菌,可以改变水体中真核微生物群落的结构,从而实现对池塘理化因子的调节。研究结果对于降低水产养殖尾水对水域环境的污染具有一定的意义。     

Long-term fertilization leads to specific PLFA finger-prints in Chinese Hapludults soil

Soil microbes play essential roles in the biogeochemical processes of organic carbon and nutrient cycling. Many studies have reported various short-term effects of fertilization on soil microbes. However, less is known about the effects of longterm fertilization regimes on the rhizosphere. Therefore, the objective of this study was to explore how the soil microbial communities in the rhizosphere respond to different long-term fertilization strategies. Based on a 21-year field treatment experiment in Guizhou, China, we extracted phospholipid fatty acids(PLFAs) to determine the microbial community structure in both the non-rhizosphere(NR) and rhizosphere(R). Six treatments were included: no fertilizer(CK), mineral nitrogen fertilizer(N), N with potassium(NK), phosphorus with K(PK), NPK, and NPK combined with manure(MNPK). The results showed that total PLFAs under unbalanced mineral fertilization(N, NK and PK) were decreased by 45% on average in the NR compared with CK, whereas MNPK increased fungi and G~– bacteria abundance significantly in both the NR(by 33 and 23%) and R(by 15 and 20%), respectively. In addition, all microbial groups in the R under these treatments(N, NK and PK) were significantly increased relative to those in the NR, except for the ratio of F/B and G~+/G~–, which might be due to the high nutrient availability in the R. Soil pH and SOC significantly regulated the soil microbial community and structure, explaining 51 and 20% of the variation in the NR, respectively. However, the rhizosphere microbial community structure was only significantly affected by soil pH(31%). We concluded that the soil microbial community in the NR was more strongly affected by long-term fertilization than that in the R due to the rhizosphere effect in the agricultural ecosystem. Rhizosphere nutrient conditions and buffering capacity could help microbial communities resist the change from the long-term fertilization.