Incidence of antibiotic-resistant Klebsiella pneumoniae and Enterobacter species in freshwater wetlands

AIMS: The aim of this study was to assess the incidence of Enterobacteriaceae (potential human and animal pathogens) in wetlands. METHODS: Enterobacteriaceae, selected from the sediments and rhizosphere of wetland plant Juncus effusus L., were analysed using classical microbiological methods, API20E, API20NE, fatty acid analyses, and 16S rRNA sequencing. Assessed virulence factors include antibiotic resistance, presence of plasmids and capsules. RESULTS: Klebsiella pneumoniae, Enterobacter cloacae and Enterobacter asburiae, known human pathogens, were identified. K. pneumoniae 16S rRNA gene sequence showed the significant hit (E < 0.001) with the unculturable bacteria obtained from faeces of elderly individuals (accession number AB099804) when Genbank database was used. Ent. asburiae 16S rRNA gene sequence showed the significant hit with (E < 0.001) with the unculturable bacteria obtained from the pig gastrointestinal tract (accession number AF371852). The rate of antibiotic resistance (<50 microg ml(-1)) was high for ampicillin and cephalosporins for the most strains (75.7%) yet low (>10 to 20 microg ml(-1)) for kanamycin, tetracycline and chloramphenicol for all strains tested. Capsules were detected in all investigated strains. PCR detected membrane protein but not chromosomally encoded beta-lactamase. Significance and Impact of the Study: The antibiotic resistance of tested strains and presence of capsules (protect micro-organisms from phagocytosis) suggest that wetland sediments and rhizosphere present a potential reservoirs for enteric human and animal pathogens.     

几种湿地植物净化生活污水COD、总氮效果比较

以无植被、基质为河砂的潜流型人工湿地为对照,研究了石菖蒲、灯心草和蝴蝶花3种类型植被、基质均为河砂的潜流型人工湿地净化生活污水COD、总氮的效果．结果表明,在污水COD浓度小于200mg·L^-1、总氮浓度小于30mg·L^-1的低浓度范围里,无植被的人工湿地和有植被的人工湿地对污水中COD、总氮均有很好的去除效果,两者差异不大,其COD去除率均达90％以上,总氮的去除率达80％以上．随着污水中COD和总氮浓度的增加,无植被人工湿地和有植被人工湿地去除COD和总氮的效果均有不同程度下降,两者差异明显,有植被的人工湿地能维持较高的COD、总氮的去除效果,无植被的人工湿地COD和总氮去除效果下降很快,植被在人工湿地系统去除污水COD和总氮过程中起着重要的作用．在整个试验阶段,石菖蒲植被人工湿地COD和总氮平均净化效率分别为80．46％和77．77％、灯心草人工湿地分别为75．53％和71．17％、蝴蝶花人工湿地分别为70．50％和66．38％,无植被人工湿地分别为61．39％和55．81％．同无植被人工湿地COD和总氮净化效果相比,石菖蒲植被人工湿地净化效果最好;其次为灯心草植被人工湿地,再次为蝴蝶花植被人工湿地．不同类型植被的人工湿地净化污水中COD和总氮的效果与其生物量关系密切,这与植被系统吸收同化有机物质和总氮数量、根际微生物分解有机物质和硝化-反硝化作用有关。     

南方红豆杉产IAA内生芽胞杆菌的分离、鉴定及产脂肽类化合物研究

通过组织研磨法对南方红豆杉(Taxus wallichiana var. mairei)新鲜组织进行内生细菌的分离，利用16S rRNA测序技术对分离的内生细菌进行鉴定分析，使用高效液相色谱-高分辨飞行质谱测定菌液提取物IAA含量，利用HPLC-TOF-MS对提取物脂肽类化合物进行分离、鉴定，结合数据库筛选菌株产脂肽类化合物种类及含量差异。结果表明，从南方红豆杉根茎叶中共分离鉴定出内生细菌31株，筛选出产IAA菌株20株，其中编号为KLBMPTC10的芽胞杆菌属菌株产IAA含量高达210.3955 μg·L^-1。在对产IAA菌株所产脂肽类化合物的研究中发现，菌株KLBMPTC10是所有产IAA内生细菌中脂肽类化合物含量最丰富的，其产Iturin A2相对含量为26.06%，明显优于其他产IAA菌株。芽胞杆菌属菌株KLBMPTC10同时具备高产IAA和脂肽类化合物的能力，可作为研究菌株进一步开发。     

Elemental composition of native wetland plants in constructed mesocosm treatment wetlands

Plants that accumulate a small percentage of metals in constructed treatment wetlands can contribute to remediation of acidic, metal contaminated runoff waters from coal mines or processing areas. We examined root and shoot concentrations of elements in four perennial wetland species over two seasons in mesocosm wetland systems designed to remediate water from a coal pile runoff basin. Deep wetlands in each system contained Myriophyllum aquaticum and Nymphaea odorata; shallow wetlands contained Juncus effusus and Pontederia cordata. Shoot elemental concentrations differed between plants of deep and shallow wetlands, with higher Zn, Al, and Fe concentrations in plants in shallow wetlands and higher Na, Mn, and P concentrations in plants in deep wetlands. Root and shoot concentrations of most elements differed between species in each wetland type. Over two seasons, these four common wetland plants did help remediate acidic, metal-contaminated runoff from a coal storage pile.     

玉米根际高效溶磷菌的筛选、鉴定及促生效应研究

为获得玉米根际高效溶磷促生菌(Plant growth-promoting rhizobacteria,PGPR)并明确其促生特性,采用选择培养方法从玉米根际土壤筛选出优良PGPR 菌株,测定其溶磷及分泌吲哚乙酸(IAA)的能力,并对优良菌株进行鉴定;采用盆栽试验研究菌株的促生作用。结果分离到2 株优良PGPR 菌株CH07和FD11,其溶磷量分别为368.5 mg/L和321.5 mg/L,产IAA量分别为30.93 mg/L和15.93 mg/L。形态学特征、生理生化特征和16S rDNA 序列分析结果表明,CH07为芽孢杆菌属(Bacillus aryabhattai),FD11为链霉菌属(Streptomyces maritimus)。最后通过盆栽试验对这2株细菌分离物的促生效果进行比较,结果发现,CH07、FD11,尤其是CH07与FD11的复合物,对苋菜的株高及地上部鲜重有积极作用,可作为研制生物肥料的优良菌株资源。     

春兰根中可分泌吲哚乙酸的内生细菌多样性

植物内生细菌可通过分泌吲哚乙酸(Indole-3-aceticacid,IAA)等方式促进植物生长。本研究以温室盆栽春兰(Cymbidium goeringii)为材料,采用分离培养方法对春兰根中可分泌IAA的内生细菌多样性进行了研究。从春兰根组织中共分离纯化得到了256株内生细菌,其中57株具有分泌IAA的能力,占总菌数的22.3%。根据ARDRA(amplified ribosomal DNA restriction analysis)及16SrDNA系统发育分析结果,将57株内生细菌划分为25个组,分属于6大类群,分别为变形菌门的α-变形菌纲(35.1%)、γ-变形菌纲(14.0%)和β-变形菌纲(8.8%)、厚壁菌门(33.3%)、放线菌门(7.0%)及拟杆菌门(1.8%)。其中变形菌门的α-变形菌纲和厚壁菌门为优势类群,类芽孢杆菌属(Paenibacillus)为优势菌属,且为高产IAA的主体菌属。另外,测序结果显示有4个菌株的序列与已知细菌的最高序列相似性低于97.0%,可能为潜在的新种或新属。研究结果表明春兰根中分泌IAA的内生细菌具有丰富的多样性。这一结果可为研究和开发植物促生细菌提供基础资料...     

Physiological responses of Juncus effusus (Rush) to chromium and relevance for wastewater treatment in constructed wetlands

Constructed wetlands are increasingly applied for industrial wastewater treatment. However, current knowledge of the stress responses of helophytes to selected toxicants such as dichromate is limited. The goal of the experiments presented here was to characterize the physiological response of Juncus effusus to different concentrations of dichromate dependent upon the growth and constitution of the plants. The growth parameters, shoot length, and dry weight already were strongly affected at low dichromate concentrations of approximately 34 microM. Concentrations of 340 microM impaired chlorophyll fluorescence and a decrease in chlorophyll a started at concentrations higher than 170 microM dichromate. The concentrations of chlorophyll b and carotenoids also were influenced negatively. Thus, the reduction of the pigment content started before any obvious influence on the chlorophyll fluorescence. The highest concentration of dichromate, which caused no permanent inhibition of growth and photosynthesis, was found to be 17 microM K2Cr2O7. In principle, J. effusus is suitable for constructed wetlands to treat chromium-containing wastewater. Because the stress resistance of J. effusus is limited, the maximum concentration of dichromate in the treated wastewater should not exceed 34 microM. The growth parameters, shoot length, and dry weight were sensitive to much lower dichromate concentrations and did react more quickly than the biochemistry-related parameters chlorophyll fluorescence and pigment concentration. Therefore, the example of Juncus effusus shows that the use of only biochemical parameters to define concentration limits for the treatment of dichromate-containing wastewater can lead to incorrect conclusions and result in disturbed long-term operation of the system.     

高产吲哚乙酸及高泌氨巴西固氨螺菌的筛选与鉴定

目的：从玉米根际和土壤中分离具有高产吲哚乙酸较强的泌氨能力的巴西固氮螺菌。方法：分别通过半固体NFb培养基、CR培养基、LB培养基分离培养固氮菌株,并经过一系列菌落菌体形态特征、生理生化特性和16S rDNA序列测定等试验对其进行鉴定。结果：经分离纯化获得10株固氮菌,并鉴定均为巴西固氮螺菌（Azospirillum brasilense）,其中菌株R7在甘油半固体培养基上能分泌约14mmol/L的氨,在添加了色氨酸的培养基中能够合成58.8μg/ml的吲哚-3-乙酸（IAA）。结论：成功筛选得到一株既高产吲哚乙酸又有较强的泌氨能力的巴西固氮螺菌。     

Interactions between Plants and Epiphytic Bacteria Regarding Their Auxin Metabolism

Using hydrocultured pea plants, 109 bacterial strains (42 from shoots) were isolated from shoots, roots, and from the hydroculture medium. 58 different strains (26 from shoots) were able to produce IAA from tryptophan, 15 different strains (7 from shoots) were able lo destroy IAA. (Included are 13 strains possessing both properties.) As far as they could be identified, the IAA-producing and -destroying strains belong to Pseudomonas (by far dominating), Achromobacter, Alcaligenses, Bacillus, and Flavobacterium. The IAA-destroying activity strongly depends on the physiological state of the bacteria and the milieu conditions. Bacterial IAA production (but not IAA-degradation) is supposed to be important for the plant.     

3种湿地植物对锌的吸收分配及其与根表铁氧化物胶膜的关系

在室内培养条件下,以灯心草、茭白和美人蕉3种湿地植物为材料,研究了湿地植物对锌的吸收分配能力与根表铁氧化物胶膜之间的关系.结果表明:(1)3种湿地植物积累锌的总量大小顺序为:茭白>美人蕉>灯心草,茭白积累锌的总量是灯心草的1.79倍;它们根表铁氧化物胶膜含量表现为灯心草>茭白>美人蕉,且其间存在显著差异(P<0.05).(2)锌在湿地植物中分配比例表现为;根中锌量>地上部分锌量>根表铁氧化物胶膜上吸附锌量;锌主要积累在湿地植物根中,地上部分和根表铁氧化物胶膜上吸附的锌量无显著差异.(3)湿地植物根表铁氧化物胶膜上吸附锌的数量与湿地植物地下部分锌含量呈极显著正相关(r=0.983 5**),增加根表铁氧化物胶膜上锌的数量就能明显提高地下部分锌含量;每千克土壤加入1 g FeSO4后,3种湿地植物积累锌的总量平均增加了21%.可见,湿地植物根表铁氧化物胶膜对锌的吸附也是湿地植物固定或积累锌的重要途径之一.     

Indole-3-acetic acid: A widespread physiological code in interactions of fungi with other organisms

Plants as well as microorganisms, including bacteria and fungi, produce indole-3-acetic acid (IAA). IAA is the most common plant hormone of the auxin class and it regulates various aspects of plant growth and development. Thus, research is underway globally to exploit the potential for developing IAA-producing fungi for promoting plant growth and protection for sustainable agriculture. Phylogenetic evidence suggests that IAA biosynthesis evolved independently in bacteria, microalgae, fungi, and plants. Present studies show that IAA regulates the physiological response and gene expression in these microorganisms. The convergent evolution of IAA production leads to the hypothesis that natural selection might have favored IAA as a widespread physiological code in these microorganisms and their interactions. We summarize recent studies of IAA biosynthetic pathways and discuss the role of IAA in fungal ecology.     

地锦草内生细菌多样性、拮抗及促生特性测定

【背景】植物内生细菌既能抑制病原菌对植物的侵染,也具有促生作用。分离具有拮抗和促生活性的内生细菌可为开发微生物菌肥提供理论依据。【目的】筛选内生细菌中的优势拮抗、促生菌种资源。【方法】以地锦草为材料,采用4种分离培养基分离该植物内生细菌,通过形态特征以及16S r RNA基因序列分析,鉴定内生细菌的分类归属。采用平板对峙法,测定内生细菌对棉花立枯丝核菌(Rhizoctonia solani)、小麦赤霉病菌(Fusarium graminearum)、玉米小斑病菌(Bipolaris maydis)的拮抗活性。通过固氮、解磷、产吲哚乙酸(Indole Acetic Acid,IAA)、产铁载体能力等指标初步检测地锦草内生细菌的促生活性。【结果】共分离到133株内生细菌,分属于4门5纲8目13科25属,其中变形菌门(Proteobacteria)为优势门(52.63%),优势属为芽孢杆菌属(Bacillus),占15.79%。发现有8株菌相似性小于98.65%,可能为潜在新物种。拮抗活性结果表明,22株菌有不同程度的抑菌作用,其中菌株DHL56、DHN17、DHP3、DHP8对这3种病原菌...     

悉生培养微缩体系食细菌线虫提高土壤激素含量的机制

研究土壤食细菌线虫与细菌的相互作用及其生态功能是土壤生态学的核心内容之一。食细菌线虫取食细菌可以促进土壤中氮素的矿化,提高氮素养分的供给,改善土壤的营养条件,从而促进植物的生长发育。土壤食细菌线虫促进植物根系生长的"养分作用机制"已得到确认,而"激素作用机制"还存在争议。从供试土壤中筛选获得一株高效产IAA细菌和两种不同cp值的食细菌线虫,通过设置简化的悉生培养系统,对这两种土著食细菌线虫与土著产IAA细菌之间的相互作用,及其对土壤中IAA含量变化的影响进行研究。结果表明:两种食细菌线虫的取食均能促进细菌数量和活性的增强,食细菌线虫与产IAA细菌相互作用也能显著增加土壤中IAA的含量;这些促进作用受到接种食细菌线虫的种类以及培养时间的影响:在培养第10天和第20天时,接种cp值为1的中杆属食细菌线虫显著增加了产IAA细菌的数量;在培养第10天和第30天时,相比较接种cp值为2的头叶属食细菌线虫,接种中杆属食细菌线虫显著提高了土壤中IAA的含量。     

Aerenchyma formation in the wetland plant Juncus effusus is independent of ethylene

Flooded plant roots commonly form aerenchyma, which allows gas diffusion between shoots and roots. The programmed cell death involved in this induced aerenchyma formation is controlled by the plant hormone ethylene, as has been shown for maize (Zea mays). However, the role of ethylene is uncertain in wetland species that form constitutive aerenchyma (also under nonflooded conditions). The aim of this study is to shed light on the involvement of ethylene in constitutive aerenchyma formation in Juncus effusus. Plants of J. effusus and maize were treated with ethylene and inhibitors of ethylene action to determine the consequences for aerenchyma formation. Neither treatment with high ethylene concentrations nor with ethylene inhibitors resulted in changes in root aerenchyma in J. effusus. By contrast, ethylene increased aerenchyma development in maize unless ethylene action inhibitors were applied simultaneously. Similarly, root elongation was insensitive to ethylene treatment in J. effusus, but was affected negatively in maize. The data show that aerenchyma in J. effusus is highly constitutive and, in contrast to the inducible aerenchyma in maize, is not obviously controlled by ethylene.     

Higher diversity of ammonia/ammonium-oxidizing prokaryotes in constructed freshwater wetland than natural coastal marine wetland

Anaerobic ammonium-oxidizing (anammox) bacteria, aerobic ammonia-oxidizing archaea (AOA), and ammonia-oxidizing bacteria (AOB) are three groups of ammonium/ammonia-oxidizing prokaryotes (AOPs) that are involved in the nitrogen cycle. This research compared the AOP communities in a constructed freshwater wetland with a natural coastal marine wetland in the subtropical Hong Kong. Both vegetated/rhizosphere and nonvegetated sediments were investigated to identify the effects of different macrophytes on the AOP communities. The polymerase chain reaction (PCR)-amplified gene fragments of 16S rRNA and archaeal and bacterial amoA (encoding the ammonia monooxygenase alpha subunit) were applied as molecular biomarkers to analyze the AOPs’ phylogeny and diversity. Quantitative PCR was used to determine the abundances of AOPs in the sediments. The results showed that the relatively more heterogeneous freshwater wetland contained a broader range of phylotypes, higher diversity, more complex community structures, and more unevenly distributed abundances of AOPs than the coastal wetland. The effects of vegetation on the community structures of AOPs were plant-specific. The exotic Typha angustifolia affected the community structures of all AOPs and enhanced their abundances in the rhizosphere region. Both Phragmites australis and Cyperus malaccensis showed some effects on the community structures of AOB, but minimal effects on those of anammox bacteria or AOA. Kandelia obovata had almost no detectable effect on all AOPs due to their smaller size. This study suggested that the freshwater and coastal marine wetlands may have different contributions to the inorganic N removal due to the variations in AOP communities and plant types.     

Wetland Restoration at a Former Nike Missile Base in the Great Lakes Basin

Restoration of an abandoned 16-ha Nike missile base site located on a former wetland in the lower Detroit River (Michigan, U.S.A.) was investigated with an emphasis on wetland restoration. The site included a 2.7-ha abandoned missile base, a 1.3-ha lake, 10.4 ha of emergent and submersed wetlands, and 1.8 ha of uplands. Aquatic beds in the shallow bay connected to the river supported floating leaved and submersed aquatics including Nymphaea tiiberosa, Vallisneria antericana, Elodea canadensis, and Heteranthera dubia, with mats of green, filamentous algae. A 10-ha diked wetland adjacent to the site was dominated by Typha spp., Salix spp., Nymphaea tuberosa, Myriophyllum spicatum, Elodea canadensis, Chara sp., and Juncus effusus. Restoration objectives included a nature preserve, an outdoor recreation area, an experimental wetland complex, and an environmental education center. Ten alternative designs were suggested, including four with wetlands open to the bay, three with diked wetlands, and the rest involving island construction, a reconstructed upland site, or a wetland research facility. Alternatives were evaluated for their contribution to local ecology, research, education and wildlife, ease of maintenance, and probability of success. Construction of a wetland resembling conditions before construction of the base was recommended for its low maintenance and opportunity for research on non diked wetland design and construction in protected bays in the Laurentian Great Lakes. Recreational and educational opportunities were also recommended as part of the site restoration.     

苎麻促生菌的筛选、鉴定及其促生效应

【目的】以苎麻(Boehmeria nivea L. Gaud)根及根围土壤为研究材料,进行苎麻促生菌的筛选,并初步探索其促生作用机制。【方法】首先,以溶磷和解钾为基本筛选标准,初筛菌株在实验室条件下测定多项促生能力进行复筛;然后通过种子萌发、盆栽试验测定菌株对苎麻的促生效应,最后,通过形态观察、生理生化特性和16S rRNA基因序列同源性分析,对促生菌株进行分类学鉴定。【结果】从苎麻根和根围土壤中分离得到了13株菌同时具备溶磷和解钾能力,其中4株菌(RA-2、RAM-2、RAM-5和RAM-6)具备产铁载体、产IAA和产氨能力。种子萌发和盆栽试验的测定结果显示：4株菌株均能促进苎麻种子的萌发和植株的生长,其中菌株RA-2和RAM-5相比于对照处理能显著提高苎麻种子的萌发率、幼根长、株高和根系干重。分类鉴定结果显示菌株RA-2和RAM-5均属于伯克霍德菌属(Burkholderia)。【结论】从苎麻根围筛选到具有促生能力的菌株,为进一步开发研制苎麻专型促生菌剂或专型微生物有机肥提供资源。     

Biofilm-associated indole acetic acid producing bacteria and their impact in the proliferation of biofilm mats in solar salterns

Biofilm mats appear in salterns distinctively during the monsoon season when the salinity decreases below 12 percentile salinity units and within a short period cover the entire surface area of the saltern. A study was carried out in two salterns viz. Nerul and Curca to find a possible reason for the rapid proliferation of these solar biofilms. Out of the 125 bacteria isolated from these biofilms, 16 produced indole-3-acetic acid (IAA). Rapid in-situ assay with Salkowski reagent and HPLC analysis confirmed the IAA production. Four isolates consistently produced high IAA concentrations ranging from 9.5 to 14.2 μg/mL in the presence of 4 mg/mL tryptophan concentrations in the growth media. The IAA-producing bacteria were Aeromonas aquariorum (N2), Pseudomonas alcaliphila (N3), Vibrio diazotrophicus (N6) and Pseudomonas pachastrellae (C3). These four IAA-producing bacteria also produced other growth promoting factors like ammonia. Three isolates produced siderophores and were phosphate solubilizers. There was enhancement in the growth of Cicer arietinum (length of the shoot and root) under axenic conditions and of biofilm mats (r = 0.9, p < 0.001; r = 0.8, p < 0.05 and r = 0.946, p < 0.01, respectively). This is, according to our knowledge, the first report indicating IAA-producing bacteria isolated from biofilms enhancing the proliferation of these biofilm mats in the solar salterns.     

花生根际微生物的分离及高效功能菌株筛选

【背景】花生根际分布着丰富的微生物类群,分离筛选多种功能的高效微生物是研发高效复合菌肥的基础。【目的】从花生根际土壤及根表分离微生物,分析可培养微生物的多样性,筛选高效解有机磷和无机磷、产吲哚乙酸(indole-3-acetic acid, IAA)和铁载体功能的菌株,为研发花生微生物菌肥打下基础。【方法】利用稀释涂布法,从采自山东省栖霞市、平度市、烟台市莱山区 3个样点的花生根际土、根表样品中分离微生物,基于16S rRNA基因序列对其进行系统发育分析,并通过初筛和复筛筛选高效解磷、产IAA和铁载体的菌株。【结果】共分离、纯化、保藏147株菌,其中75株分离自根际土壤,72株分离自根表样品。系统发育分析表明所有的菌分布于放线菌门(Actinomycetota)、芽孢杆菌门(Bacillota)、拟杆菌门(Bacteroidota)和假单胞菌门(Pseudomonadota)这4个门的40个属,优势属为链霉菌属(Streptomyces, 21.77%)、芽孢杆菌属(Bacillus, 16.33%);根表样品微生物的多样性高于根际样品;共筛选到解有机磷菌株62株,短波单胞菌(Brevundimonas) YTU21021解有机磷能力最强为1.12 mg/L;解无机磷菌株31株,不动杆菌(Acinetobacter) YTU21009解无机磷能力最强为7.04 mg/L;产IAA的菌株63株,肠杆菌(Enterobacter) YTU21054产IAA量最高,达184.19 mg/L;产铁载体细菌7株,伯克氏菌(Burkholderia) YTU21051产铁载体能力最强,A_s/A_r为0.90。【结论】花生根际和根表样品中可培养微生物多样性较为丰富,本研究筛选到的高效功能菌丰富了花生根际功能微生物资源,为后续与高效根瘤菌联合研发花生复合微生物菌肥奠定了基础。     

乌头产吲哚乙酸内生细菌遗传多样性、抗逆性及其对水稻幼苗生长的影响

[目的]探究乌头产吲咮乙酸(IAA)内生细菌的遗传多样性、溶磷解钾能力、抗逆能力及其对水稻幼苗生长的影响,为道地产区乌头产业可持续发展提供科技支撑.[方法]从健康乌头植株分离可培养内生细菌,采用Salkowski比色法测定内生细菌产IAA能力,16S rDNA限制性片段长度多样性(16S rDNA-RFLP)及16S ...