Anaerobic nitrate reduction to ammonium in two strains isolated from coastal marine sediment: A dissimilatory pathway

Abstract: A total of 28 nitrate-reducing bacteria were isolated from marine sediment (Mediterranean coast of France) in which dissimilatory reduction of nitrate to ammonium (DRNA) was estimated as 80% of the overall nitrate consumption. Thirteen isolates were considered as denitrifiers and ten as dissimilatory ammonium producers. ¹⁵N ammonium production from ¹⁵N nitrate by an Enterobacter sp. and a Vibrio sp., the predominant bacteria involved in nitrate ammonification in marine sediment, was characterized in pure culture studies. For both strains studied, nitrate-limited culture (1 mM) produced ammonium as the main product of nitrate reduction (> 90%) while in the presence of 10 mM nitrate, nitrite was accumulated in the spent media and ammonia production was less efficient. Concomitantly with the dissimilation of nitrate to nitrite and ammonium the molar yield of growth on glucose increased. Metabolic products of glucose were investigated under different growth conditions. Under anaerobic conditions without nitrate, ethanol was formed as the main product; in the presence of nitrate, ethanol disappeared and acetate increased concomitantly with an increased amount of ammonium. These results indicate that nitrite reduction to ammonium allows NAD regeneration and ATP synthesis through acetate formation, instead of ethanol formation which was favoured in the absence of nitrate.     

Biochemical characterization of some cyanobacterial strains from salt marshes of the Venice Lagoon

Three different strains of filamentous cyanobacteria, Tychonema, Limnothrix, and Pseudoanabaena, were selected among the fastest growing taxa collected in the salt marshes of Venice Lagoon and were grown in laboratory for growth rate determination and biochemical characterization of chlorophyll-a, total proteins, total carbohydrates, and exopolysaccharides. Experiments were carried out both in liquid medium and two different substrates: artificial plant protection fabric and ground indigenous shells. Cyanobacterial behavior was recorded to better understand colonization of natural and new artificial marshes.     

Zearalenone degradation by two Pseudomonas strains from soil

This study describes the screening of soil bacteria for their capability to degrade zearalenone (ZEN), employing an enrichment technique in which ZEN is used as the sole carbon source. Two isolates that were able to degrade ZEN belonged to the genus Pseudomonas according to biochemical characterization and 16S rRNA gene sequence and were named as Pseudomonas alcaliphila TH-C1 and Pseudomonas plecoglossicida TH-L1, respectively. The results showed that the degradation rates of P. alcaliphila TH-C1 and P. plecoglossicida TH-L1 for ZEN (2 μg/ml) were 68?±?0.85 % and 57?±?0.73 %, when incubated for 72 h at 30 °C in a rotary shaker (160 rpm) and detected by high-performance liquid chromatograms (HPLC). These results suggest that the two Pseudomonas strains are new bacterial resource for degrading ZEN and can provide a new approach for the detoxification of ZEN.     

Occurrence of antibiotic and metal resistance and plasmids in Bacillus strains isolated from marine sediment

Eleven hundred Bacillus strains isolated from marine sediment from the Minas Basin, Nova Scotia, Canada, were purified on LB agar supplemented with ampicillin, chloramphenicol, erythromycin, streptomycin, tetracycline, or mercuric chloride. Seventy-seven isolates were examined for plasmid DNA, and for resistance to 11 antibiotics, HgCl2, and phenylmercuric acetate. Minimum inhibitory concentrations of Ag, Cd, Co, Cu, and Zn were also determined. Forty-three percent of antibiotic- and mercury-resistant strains contained one or more plasmids ranging from 1.9 to 210 MDa. Fifty-four percent carried plasmids greater than 20 MDa, and 97% were resistant to two or more metals. There was no correlation between plasmid content and resistance either to antibiotics or to mercurial compounds in these strains. Mercury-resistant isolates were unable to transform Hg2+ to volatile Hg0 by virtue of a mercuric reductase enzyme system (mer). Strains resistant to Hg2+ were investigated for their ability to produce H2S and intracellular acid-labile sulfide when grown in the absence and presence of HgCl2. Lower levels of H2S and intracellular sulfide were detected only in metal-resistant strains grown in the presence of HgCl2, suggesting that cellular sulfides complexed with Hg2+ in these strains.     

Naphthalene oxidation by a Pseudomonas putida strain carrying a mutant plasmid

Naphthalene oxidation by a parent and a mutant strain of Pseudomonas putida was studied. The parent strain contained a plasmid NPL-1 which controlled oxidation of naphthalene to salicylic acid and was capable of oxidizing salicylate. The mutant strain did not oxidize salicylate because of a mutation in salicylate hydroxylase; it contained also a mutant plasmid NPL-41 which determined constitutive synthesis of naphthalene oxygenase. Salicylic acid which accumulated as a product of naphthalene catabolism in the cultural broth of the wild strain was found to undergo further oxidation by the population of growing cells. The content of salicylic acid in the cultural broth of the mutant strain reached maximum and then remained constant. An anion-exchange resin was tested in order to prevent the inhibition of naphthalene oxygenase by salicylate and to increase the yield of salicylic acid. The transmissible character of the mutant plasmid NPL-41 makes it possible, with the aid of conjugation, to construct Pseudomonas strains which would oxidize naphthalene to salicylic acid without further degradation of this compound.     

Tetrodotoxin production of actinomycetes isolated from marine sediment

Ten actinomycetes isolated from various marine sediments were investigated for the production of tetrodotoxin (TTX). Tissue culture assay, high performance liquid chromatography and gas chromatography-mass spectrometry confirmed that the nine strains produced TTX. Actinomycetes may be responsible for TTX accumulation in marine environments.     

Aerobic and anaerobic metabolism of squalene by a denitrifying bacterium isolated from marine sediment

The aerobic and anaerobic metabolism of the isoprenoid alkene squalene was investigated in a new type of marine denitrifying bacterium, strain 2sq31, isolated from marine sediment. Strain 2sq31 was identified as a species of Marinobacter. Under denitrifying conditions, the strain efficiently degraded squalene; of 0.7 mmol added per liter of medium, 77% was degraded within 120 days under anoxic conditions with nitrate as electron acceptor. Tertiary diols and methyl ketones were identified as metabolites, and an anaerobic pathway was suggested to explain the formation of such compounds. The first step in anaerobic degradation of squalene by strain 2sq31 involves hydration of double bonds to tertiary alcohols. Under oxic conditions, the degradation of squalene by strain 2sq31 was rapid and involved oxidative splitting of the C-10/C-11 or C-14/C-15 double bonds, in addition to the pathways observed under denitrifying conditions.     

两株假单胞菌对吡啶和喹啉的生物降解

【目的】探究2株假单胞菌(Pseudomonas)对吡啶和喹啉的降解。【方法】基于16S rRNA序列同源性和基因间区分析,对分离菌株进行分类鉴定。通过分光光度法和电喷雾电离质谱法(Electrospray Ionisation/Mass Spectrometry,ESI/MS)确定分离菌株对吡啶和喹啉的降解性能。通过质粒消除验证降解质粒的存在,同时克隆了可能的降解基因。【结果】鉴定结果表明,两株分离细菌隶属于Pseudomonas,并将其命名为XJUHX-1和XJUHX-12。降解数据表明,2株菌株分别耐受吡啶和喹啉,同时分别检测到4种和2种吡啶和喹啉的可能降解产物。结果还表明,消除质粒后的菌株对吡啶和喹啉的降解能力降低。扩增的编码NADH还原酶部分的降解喹啉oxoR基因和编码硝酸还原酶的降解吡啶的nifH基因,同时在E.coli中表达了43kDa和16kDa的蛋白。【结论】2株Pseudomonas具有降解吡啶和喹啉的能力。     

Metal-dependent anaerobic methane oxidation in marine sediment: Insights from marine settings and other systems

Anaerobic oxidation of methane(AOM) plays a crucial role in controlling global methane emission. This is a microbial process that relies on the reduction of external electron acceptors such as sulfate, nitrate/nitrite, and transient metal ions. In marine settings, the dominant electron acceptor for AOM is sulfate, while other known electron acceptors are transient metal ions such as iron and manganese oxides. Despite the AOM process coupled with sulfate reduction being relatively well characterized,researches on metal-dependent AOM process are few, and no microorganism has to date been identified as being responsible for this reaction in natural marine environments. In this review, geochemical evidences of metal-dependent AOM from sediment cores in various marine environments are summarized. Studies have showed that iron and manganese are reduced in accordance with methane oxidation in seeps or diffusive profiles below the methanogenesis zone. The potential biochemical basis and mechanisms for metal-dependent AOM processes are here presented and discussed. Future research will shed light on the microbes involved in this process and also on the molecular basis of the electron transfer between these microbes and metals in natural marine environments.     

Hyphomonas sediminis sp. nov., isolated from marine sediment

A Gram-staining-negative, aerobic and pear-shaped bacterial strain, designated WL0036^T, was isolated from coastal sediment sample collected in Nantong city, Jiangsu province of China (120° 51′ 13″ E, 32° 6′ 26″ N) in October 2020. Strain WL0036^T was found to grow at 20–37 °C (optimum, 28 °C) with 0–9.0% NaCl (optimum, 2.5–4.0%) and displayed alkaliphilic growth with the pH range of pH 6.0–10.0 (optimum, pH 7.0–8.0). The polar lipids profile of strain WL0036^T included phosphatidylcholine, phosphatidylethanolamine, glycolipid and an unidentified lipid. The major isoprenoid quinone was determined to be Q-11 and the major fatty acids were C_16:0, 11-methyl-C_18:1ω7c, and summed features 8 (C_18:1ω6c and/or C_18:1ω7c). The G?+?C content of genomic DNA was 61.8%. Phylogenetic trees constructed based on 16S rRNA gene sequence and bac120 gene set (a collection of 120 single-copy protein sequences prevalent in bacteria) indicted that strain WL0036^T clustered with strains Hyphomonas neptunium ATCC 15444^T and H. polymorpha PS728^T. The average nucleotide identities between strain WL0036^T and strains H. neptunium ATCC 15444^T and H. polymorpha PS728^T were 80.7% and 81.2%, respectively. Strain WL0036^T showed 22.8% and 23.2% of digital DNA-DNA hybridization identities with H. neptunium ATCC 15444^T and H. polymorpha PS728^T, respectively. As inferred from the phenotypic and genotypic characteristics and the phylogenetic trees, strain WL0036^T ought to be recognized as a novel species in genus Hyphomonas, for which the name Hyphomonas sediminis sp. nov. is proposed. The type strain is WL0036^T (=?MCCC 1K05843^T?=?JCM 34658^T?=?GDMCC 1.2413^T).

     

Fictibacillus enclensis sp. nov., isolated from marine sediment

A novel Gram-positive strain, designated NIO-1003^T, was isolated from a marine sediment sample collected from the Chorao Island, Goa Provence, India. Strain NIO-1003^T was found to be strictly aerobic, motile, endospore-forming rods. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NIO-1003^T belongs to the genus Fictibacillus and to be most closely related to Fictibacillus rigui KCTC 13278^T, Fictibacillus solisalsi KCTC 13181^T and Fictibacillus barbaricus DSM 14730^T with 98.2, 98.0 and 97.2 % similarity and 25, 28, 39 nucleotide differences respectively. Strain NIO-1003^T was characterized by having cell-wall peptidoglycan based on meso-diaminopimelic acid and MK-7 as the predominant menaquinone. The polar lipid profile exhibited the major compounds diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. In addition, minor amounts of an aminophospholipid were detected. The major fatty acids were identified as ai-C_15:0, iso-C_15:0, ai-C_17:0 and C_16:0, supporting the grouping of strain NIO-1003^T into the family Bacillaceae. The DNA G+C content of strain NIO-1003^T was determined to be 42.6 mol%. On the basis of phenotypic properties, phylogeny and DNA–DNA hybridisation analysis, strain NIO-1003^T is considered to represent a novel species of the genus Fictibacillus for which the name Fictibacillus enclensis sp. nov. is proposed. The type strain is NIO-1003^T (= NCIM 5458^T = DSM 25142^T).     

Siderophore synthesis by mucoid Pseudomonas aeruginosa strains isolated from cystic fibrosis patients.

Nonmucoid Pseudomonas aeruginosa responds to iron deprivation by synthesizing the siderophores pyochelin and pyoverdine. When grown in iron-deficient medium, six mucoid P. aeruginosa strains isolated from cystic fibrosis patients synthesized copious amounts of the exopolysaccharide alginate. A procedure that eliminated the interference of alginate was developed so that siderophores could be extracted from the growth medium. All six isolates were then noted to produce both pyoverdine and pyochelin. This report thus confirms that mucoid P. aeruginosa, like its nonmucoid counterparts, elicits the siderophores commonly cited as those of the microbe.     

Fluoroquinolone susceptibility of Pseudomonas aeruginosa strains isolated from clinical materials

The goal of our research was an analysis of sensitivity of Pseudomonas aeruginosa to fluoroquinolones (SPX, PEF, UB, LOM, CIP, ENX, OFX, NOR). The sensitivity was tested by disk diffusion method, according to NCCLS standards. 120 strains isolated from hospitalized (76 strains) and outpatient clinic (44 strains) persons were tested. The highest sensitivity of strains was observed to norfloxacin (36.8% and 86.4% of strains, respectively) and ciprofloxacin (30.3% and 81.8%). None of tested microorganisms was sensitive to flumequine. Resistant strains, isolated from sick persons in outpatient clinics were fewer (13.6%) as compared to hospitalized persons (51.3%).     

Production of acylated homoserine lactones by Aeromonas and Pseudomonas strains isolated from municipal activated sludge

Up to now, the production and role of N-acyl homoserine lactones (AHLs) in activated sludge have been poorly understood. In this study, cross-feeding assays with the reporter strains Agrobacterium tumefaciens NTL4 and Chromobacterium violaceum CV026 were used to investigate AHL signal production by municipal activated sludge samples. AHL signal production was consistently detected from municipal activated sludge when different samples were incubated on nutrient media. From one municipal activated sludge sample, 10 strains producing AHL-like auto inducers were isolated by an overlay technique. 16S rDNA-based phylogenetic analysis showed that eight of the isolates belonged to Aeromonas spp. and two to Pseudomonas spp. Box-PCR indicated that six of these Aeromonas isolates were different strains and the two Pseudomonas strains were identical. The production of AHL or AHL-like compounds by these strains was confirmed by thin layer chromatography and biosensor overlays. The six different Aeromonas strains were found to produce the same set of AHLs, including N-hexanoyl-L-homoserine lactone. These results may indicate the possible presence of AHLs in municipal activated sludge. The potential roles of AHL in this eco system are briefly discussed.