Influence of an oil soluble inhibitor on microbiologically influenced corrosion in a diesel transporting pipeline

Abstract

Microbial degradation of the oil soluble corrosion inhibitor (OSCI) Baker NC 351 contributed to a decrease in inhibitor efficiency. Corrosion inhibition efficiency was studied by the rotating cage and flow loop methods. The nature of the biodegradation of the corrosion inhibitor was also analysed using Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry. The influence of bacterial activity on the degradation of the corrosion inhibitor and its influence on corrosion of API 5LX were evaluated using a weight loss technique and impedance studies. Serratia marcescens ACE2 and Bacillus cereus ACE4 can degrade aromatic and aliphatic hydrocarbons present in the corrosion inhibitor. The present study also discusses the demerits of the oil soluble corrosion inhibitors used in petroleum product pipeline.     

Role of <Emphasis Type="Italic">Serratia marcescens</Emphasis> ACE2 on diesel degradation and its influence on corrosion

A facultative anaerobic species Serratia marcescens ACE2 isolated from the corrosion products of diesel transporting pipeline in North West, India was identified by 16S rDNA sequence analysis. The role of Serratia marcesens ACE2 on biodegradation of diesel and its influence on the corrosion of API 5LX steel has been elucidated. The degrading strain ACE2 is involved in the process of corrosion of steel API 5LX and also utilizes the diesel as an organic source. The quantitative biodegradation efficiency (BE) of diesel was 58%, calculated by gas-chromatography–mass spectrum analysis. On the basis of gas-chromatography–mass spectrum (GC–MS), Fourier Transform infrared spectroscopy (FTIR) and X-ray diffractometer (XRD), the involvement of Serratia marcescens on degradation and corrosion has been investigated. This basic study will be useful for the development of new approaches for detection, monitoring and control of microbial corrosion.     

Bacterial Degradation and Corrosion of Naphtha in Transporting Pipeline

Five naphtha hydrocarbon-degrading bacteria including representative strains of the two classified species (Serratia marcescensAR1, Bacillus pumilusAR2, Bacillus carboniphilus AR3, Bacillus megaterium AR4, and Bacillus cereus AR5) were identified by 16S rDNA gene sequence in a naphtha-transporting pipeline. The naphtha-degrading strains were able to be involved in the corrosion process of API 5LX steel and also utilized the naphtha as the sole carbon source. The biodegradation of naphtha by the bacterial isolates was characterized by gas chromatography-mass spectrometry. Weight-loss measurement on the corrosion of API 5LX steel in the presence/absence of consortia grown in naphtha-water aqueous media was performed. The scanning electron microscope observation showed that the consortia were able to attack the steel API 5LX surface, creating localized corrosion (pit). The biodegradation of naphtha by the strains AR1, AR2, AR3, AR4, and AR5 showed biodegradation efficiency of about 76.21, 67.20, 68.78, 68.78, and 68.15, respectively. The role of degradation on corrosion has been discussed. This basic study will be useful for the development of new approaches for the detection, monitoring, and control of microbial corrosion in a petroleum product pipeline.     

Biodegradation and corrosion behaviour of <Emphasis Type="Italic">Serratia marcescens</Emphasis> ACE2 isolated from an Indian diesel-transporting pipeline

A facultative anaerobic species Serratia marcescens ACE2 isolated from the corrosion products of a diesel-transporting pipeline in North West India was identified by 16S rDNA sequence analysis. The role of Serratia marcesens ACE2 on biodegradation of commercial corrosion inhibitor (CCI) and its influence on the corrosion of API 5LX steel has been enlightened. The degrading strain ACE2 is involved in the process of corrosion of steel API 5LX and also utilizes the inhibitor as organic source. The quantitative biodegradation efficiency of corrosion inhibitor was 58%, which was calculated by gas chromatography mass spectrum analysis. The effect of CCI on the growth of bacteria and its corrosion inhibition efficiency were investigated. Additionally, the role of this bacterium in corrosion of steel has been investigated by powder X-ray diffractometer (XRD) and scanning electron microscope studies. The presence of high-intensity ferric oxides and manganese oxides noticed from the XRD indicates that ACE2 enhances the corrosion process in presence of inhibitor as a carbon source. This basic study will be useful for the development of new approaches for the detection, monitoring and control of microbial corrosion in petroleum product pipelines.     

Antibacterial activity of stevioside towards food‐borne pathogenic bacteria

This study determines the inhibitory effect of Stevia rebaudiana leaf extracts and its purified bioactive compound ‘stevioside’ against food‐related pathogens. The S. rebaudiana solvent extracts (1000 μg/mL) displayed antibacterial activity to Serratia marcescens, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, B. subtilis, Alcaligenes denitrificans and Salmonella typhimurium. Of the six solvents, ethanol and acetone extracts displayed the highest zone of inhibition. The bioactive compound from S. rebaudiana was purified by solvent extraction, thin‐layer chromatography followed by structural characterization by spectroscopy evidence. Purified stevioside prevented the growth of tested bacterial species, i.e. B. subtilis, K. pneumoniae and S. typhimurium. Significant zone of inhibition (12 mm) was observed against B. cereus which proposes potential application of stevioside in foods to increase their shelf life.     

Sustainable management strategies for bacterial wilt of sweet peppers (Capsicum annuum) and other Solanaceous crops

Pepper bacterial wilt is caused by the bacterial pathogen, Ralstonia solanacearum. It is the most destructive disease of many Solanaceous crops such as potatoes, tobacco, pepper, tomatoes and eggplant and is a significant source of crop loss worldwide. Physical, cultural and chemical controls have been employed to combat this destructive disease. However, none of these strategies has been able to control the disease completely due to the broad host range and genetic diversity of the pathogen, its prolonged survival in the soil and survival on vegetation as a latent infection. Owing to co-management strategies, biological control is the best approach for human health and environmental friendly motivations. It makes use of various antagonistic rhizobacteria and epiphytic species such as Bacillus cereus, Pseudomonas putida, Bacillus subtilis, Paenibacillus macerans, Serratia marcescens, Bacillus pumilus and Pseudomonas fluorescens, which compete with and ultimately inhibit the growth of the pathogen. The possible mechanisms of biocontrol by these species involve multifaceted interactions between the host, pathogen and the antagonists. These can involve competition for nutrients and space, plant-mediated systemic resistance, siderophore production and production of extracellular cell wall degrading enzymes to inhibit or suppress the growth of the bacterial wilt agent.     

Phylogenetic characterization of bacterial consortia obtained of corroding gas pipelines in Mexico

Characterization of the microbial populations formed in gas pipelines is essential to understand the metallic surface-microbe interaction, their role in metal corrosion, and to implement efficient monitoring and control strategies. Microbial community analysis in a corroded gas pipeline in a petroleum-producing facility in the Southeast region in Mexico was performed by traditional cultivation techniques and identification based on 16S rRNA gene sequence. In all samples, thin bacterial biofilms were observed and pitting corrosion was reveled after removing the biofilms. Six pure or mixed cultures of anaerobic bacteria were obtained and their 16S rRNA libraries were constructed, respectively. At least two members of each RFLP profile were sequenced and the phylogenetic affiliations of cloned bacterial 16S rRNA genes indicated that native biofilms were mainly colonized by Desulfovibrio vulgaris and Desulfovibrio desulfuricans, sulfate-reducing bacteria members; Citrobacter freundii, an Enterobacteriaceae member; Clostridium celerecrescens and Clostridium sporogenes, spore-forming anaerobic species and Cetobacterium somerae, a microaerotolerant, non-spore-forming fusobacteria. Some of these species have been observed consistently in other steel pipelines previously, but Cetobacterium members and C. celerecrescens are described for the fist time in this corroded gas pipeline. The potential role of each species in biofilm formation and steel corrosion is discussed.     

Isolation screening and characterisation of local beneficial rhizobacteria based upon their ability to suppress the growth of Fusarium oxysporum f. sp. radicis-lycopersici and tomato foot and root rot

Local beneficial rhizobacteria were selected based upon their ability to control the fungus Fusarium oxysporum f. sp. radicis-lycopersici which causes crown and root rot of tomato. Seven out of 384 strains prevailed in multiple and dual cultures and were identified as Pseudomonas chlororaphis (one strain), Bacillus cereus (one strain), Serratia marcescens (three strains) and Serratia rubidaea (two strains), by sequencing the 16S rRNA or the 16S and 23S rRNA inter-spacer region. The seven selected rhizobacteria were tested for their biocontrol and growth-promoting effects in planta, and their antifungal properties in vitro. All strains significantly reduced disease severity under controlled conditions, in a gnotobiotic system and in pots. Moreover, one P. chlororaphis and one S. marcescens strain significantly decreased disease severity to the level of the healthy control under natural conditions in pots experiments. The inhibitory activity of bacterial liquid cultures' metabolites on the fungus was demonstrated for all strains in vitro, using filter paper, thin layer chromatography and microtiter bioassays. Genes encoding phenazines were tentatively detected by PCR in the P. chlororaphis strain and chitinase-encoding genes were detected in one S. rubidaea and all three S. marcescens strains. Production of phenazine-1-carboxamide and hydrogen cyanide was evidenced for the P. chlororaphis strain while protease activity and production of siderophore-like compounds was confirmed in all bacterial strains. Possible use of these strains as biological control agents and their impact on natural biocontrol of pathogens in soils is discussed.     

The plant growth-promoting rhizobacterium Bacillus cereus AR156 induces resistance in tomato with induction and priming of defence response

In a previous study, we demonstrated the ability of the rhizobacterium Bacillus cereus AR156 (AR156) to protect tomato against bacterial wilt caused by Ralstonia solanacearum and root-knot disease caused by Meloidogyne incognita. Here, we investigate the ability of AR156 to promote plant growth and its role in the systemic protection of tomatoes cultivated in greenhouses against bacterial speck disease caused by Pseudomonas syringae pv. tomato DC3000 (DC3000). In our experiments, the AR156 population reached 10⁵–10⁶ CFU/g rhizosphere soil, and remained at that level in the rhizosphere of tomato plants for more than 2 months. In terms of its ability to promote plant growth, AR156 increased the average biomass of the tomato by 47.7%. AR156 also elicited induced systemic resistance against DC3000, significantly reduced bacterial speck disease severity 1.6-fold, and inhibited proliferation of the pathogen by approximately 15-fold. This strain triggered the accumulation of defence-related genes (PR1 and PIN2) in tomato leaves and primed the leaves for accelerated defence-related gene expression upon challenge with DC3000. That suggested simultaneous activation of the salicylic acid and the jasmonic acid dependent signalling pathways by AR156 against DC3000. In conclusion, B. cereus AR156 was found to form robust colonies in the roots of tomato and had some beneficial effects, including biological control of bacterial speck disease via ISR and promotion of plant growth.     

Characterization of Microbial Communities in Gas Industry Pipelines

Culture-independent techniques, denaturing gradient gel electrophoresis (DGGE) analysis, and random cloning of 16S rRNA gene sequences amplified from community DNA were used to determine the diversity of microbial communities in gas industry pipelines. Samples obtained from natural gas pipelines were used directly for DNA extraction, inoculated into sulfate-reducing bacterium medium, or used to inoculate a reactor that simulated a natural gas pipeline environment. The variable V2-V3 (average size, 384 bp) and V3-V6 (average size, 648 bp) regions of bacterial and archaeal 16S rRNA genes, respectively, were amplified from genomic DNA isolated from nine natural gas pipeline samples and analyzed. A total of 106 bacterial 16S rDNA sequences were derived from DGGE bands, and these formed three major clusters: beta and gamma subdivisions of Proteobacteria and gram-positive bacteria. The most frequently encountered bacterial species was Comamonas denitrificans, which was not previously reported to be associated with microbial communities found in gas pipelines or with microbially influenced corrosion. The 31 archaeal 16S rDNA sequences obtained in this study were all related to those of methanogens and phylogenetically fall into three clusters: order I, Methanobacteriales; order III, Methanomicrobiales; and order IV, Methanosarcinales. Further microbial ecology studies are needed to better understand the relationship among bacterial and archaeal groups and the involvement of these groups in the process of microbially influenced corrosion in order to develop improved ways of monitoring and controlling microbially influenced corrosion.     

Bacteria associated with Amblyomma cajennense tick eggs

Ticks represent a large group of pathogen vectors that blood feed on a diversity of hosts. In the Americas, the Ixodidae ticks Amblyomma cajennense are responsible for severe impact on livestock and public health. In the present work, we present the isolation and molecular identification of a group of culturable bacteria associated with A. cajennense eggs from females sampled in distinct geographical sites in southeastern Brazil. Additional comparative analysis of the culturable bacteria from Anocentor nitens, Rhipicephalus sanguineus and Ixodes scapularis tick eggs were also performed. 16S rRNA gene sequence analyses identified 17 different bacterial types identified as Serratia marcescens, Stenotrophomonas maltophilia, Pseudomonas fluorescens, Enterobacter spp., Micrococcus luteus, Ochrobactrum anthropi, Bacillus cereus and Staphylococcus spp., distributed in 12 phylogroups. Staphylococcus spp., especially S. sciuri, was the most prevalent bacteria associated with A. cajennense eggs, occurring in 65% of the samples and also frequently observed infecting A. nitens eggs. S. maltophilia, S. marcescens and B. cereus occurred infecting eggs derived from specific sampling sites, but in all cases rising almost as pure cultures from infected A. cajennense eggs. The potential role of these bacterial associations is discussed and they possibly represent new targets for biological control strategies of ticks and tick borne diseases.     

Influence of inoculation with plant growth promoting rhizobacteria (PGPR) on tomato plant growth and nematode reproduction under greenhouse conditions

Numerous species of soil bacteria which flourish in the rhizosphere of plants or around plant tissues stimulate plant growth and reduce nematode population by antagonistic behavior. These bacteria are collectively known as PGPR (plant growth promoting rhizobacteria). The effects of six isolates of PGPR Pseudomonas putida, Pseudomonas fluorescens, Serratia marcescens, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus cereus, were studied on tomato plant growth and root knot nematode reproduction after 45 days from nematode infection. The highest number of shoot dry weight/g (43.00 g) was detected in the plant treated with S. marcescens; then P. putida (34.33 g), B. amyloliquefaciens (31.66 g), P. fluorescens (30.0 g), B. subtilis (29.0 g), B. cereus (27.0 g) and nematode alone (untreated) 20 g/plant. While the highest number of plant height was observed when plant was treated with S. marcescens, P. fluorescens, P. putida, B. amyloliquefaciens and P. putida 52.66, 50.66, 48 and 48 cm respectively. No significant differences were seen between previous treatments but only had significant differences compared with untreated plant. The highest number of fruit/plant was observed when plants were treated with S. marcescens (10.66), then B. amyloliquefaciens (8.66), P. putida (8), P. fluorescens (8) and B. cereus (7.66). No significant differences between the last 4 treatments, but all had significant differences compared with untreated plants. The highest weight of plant yield (g) was observed with S. marcescens (319.6 g/plant) and the lowest weight of plant yield was observed in plants treated with nematode alone (untreated). On the other hand, the lowest numbers of J₂/10 g of soil (78), galls/root, (24.33) galls/root, egg masses/root (12.66) and egg/egg masses were observed in the plants treated with S. marcescens.     

Microbiological induced corrosion of AA 6061 nuclear alloy in highly diluted media by Bacillus cereus RE 10

Microbiological studies of a spent nuclear fuel pool in Argentina were performed to evaluate the risk of microbiological induced corrosion and determine the cultivable bacterial population. Based on standard methods and sequencing of the 16sRNA gene, eighteen microorganisms were identified. Bacillus cereus RE 10 was the predominant organism isolated, and was selected to investigate the biofilm formation process and the corrosion effect on aluminum alloy AA 6061 and on pure aluminum (Al 99.999%). To simulate the environmental conditions, the experiments were performed using a highly diluted medium. After 20 days of exposure, major pits covered with deposits were found on AA 6061 samples exposed to B. cereus RE 10 but not on Al 99.999%. There was a close correlation between biofilm patches, corrosion deposits, pitting and Al-(Fe or Ti)-Si inclusions. We postulate that this correlation is a consequence of a light local alkalinization around the inclusions that produces changes in the expression pattern of B. cereus RE 10 and allows bacterial survival using other substrates. Under these conditions, the generated biofilm induces a crevice corrosion effect around the intermetallic inclusions of the alloy. Our results will be useful for further studies related to the microbial impact on nuclear safety in nuclear waste storage facilities in Argentina.     

Culturable Bacteria Present in the Fluid of the Hooded-Pitcher Plant <Emphasis Type="Italic">Sarracenia Minor</Emphasis> Based on 16S rDNA Gene Sequence Data

The culturable microbial community within the pitcher fluid of 93 Sarracenia minor carnivorous plants was examined over a 2-year study. Many aspects of the plant/bacterial/insect interaction within the pitcher fluid are minimally understood because the bacterial taxa present in these pitchers have not been identified. Thirteen isolates were characterized by 16S rDNA sequencing and subsequent phylogenetic analysis. The Proteobacteria were the most abundant taxa and included representatives from Serratia, Achromobacter, and Pantoea. The Actinobacteria Micrococcus was also abundant while Bacillus, Lactococcus, Chryseobacterium, and Rhodococcus were infrequently encountered. Several isolates conformed to species identifiers (>98% rDNA gene sequence similarity) including Serratia marcescens (isolates found in 27.5% of pitchers), Achromobacter xylosoxidans (37.6%), Micrococcus luteus (40.9%), Bacillus cereus (isolates found in 10.2%), Bacillus thuringiensis (5.4%), Lactococcus lactis (17.2%), and Rhodococcus equi (2.2%). Species–area curves suggest that sampling efforts were sufficient to recover a representative culturable bacterial community. The bacteria present represent a diverse community probably as a result of introduction by insect vectors, but the ecological significance remains under explored.     

Bioaccumulation of cerium and neodymium by <Emphasis Type="Italic">Bacillus cereus</Emphasis> isolated from rare earth environments of Chavara and Manavalakurichi,India

Rare earth elements (REEs) are among the common minerals in the Rare earth environment that are very precious and also enhance soil properties. The aim of this present study is to evaluate the accumulation of REEs by bacterial isolates of rare earth environment. Morphological and biochemical characterization were done for 37 bacterial isolates and also molecular studies were carried out using 16S rRNA sequencing method. The assessment of REEs composition in soil samples of Chavara and Manavalakurichi analyzed using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) showed the abundance of Cerium and Neodymium among lanthanides. The bioaccumulation study of rare earth elements by Bacillus cereus were accomplished employing FT-IR spectrum and ICP-OES analysis. The significant accumulation of rare earth elements especially Cerium and Neodymium was noticed in Bacillus cereus isolated from rare earth environment.     

Characterization of microbial communities in gas industry pipelines

Culture-independent techniques, denaturing gradient gel electrophoresis (DGGE) analysis, and random cloning of 16S rRNA gene sequences amplified from community DNA were used to determine the diversity of microbial communities in gas industry pipelines. Samples obtained from natural gas pipelines were used directly for DNA extraction, inoculated into sulfate-reducing bacterium medium, or used to inoculate a reactor that simulated a natural gas pipeline environment. The variable V2-V3 (average size, 384 bp) and V3-V6 (average size, 648 bp) regions of bacterial and archaeal 16S rRNA genes, respectively, were amplified from genomic DNA isolated from nine natural gas pipeline samples and analyzed. A total of 106 bacterial 16S rDNA sequences were derived from DGGE bands, and these formed three major clusters: beta and gamma subdivisions of Proteobacteria and gram-positive bacteria. The most frequently encountered bacterial species was Comamonas denitrificans, which was not previously reported to be associated with microbial communities found in gas pipelines or with microbially influenced corrosion. The 31 archaeal 16S rDNA sequences obtained in this study were all related to those of methanogens and phylogenetically fall into three clusters: order I, Methanobacteriales; order III, Methanomicrobiales; and order IV, Methanosarcinales: Further microbial ecology studies are needed to better understand the relationship among bacterial and archaeal groups and the involvement of these groups in the process of microbially influenced corrosion in order to develop improved ways of monitoring and controlling microbially influenced corrosion.     

Assessment of antibacterial effects of flavonoids by estimation of generation times in liquid bacterial cultures

Antibacterial activities of various flavonoids, a group of natural plant substances, have been reported previously, however, there are contradictory data, published by various authors, regarding sensitivity of particular bacterial species to these compounds. These problems arose apparently because of using different methods by various researchers. Here we tested sensitivity of several bacterial species (Gram-positive: Bacillus subtilis, Micrococcus luteus, Sarcina sp. and Staphylococcus aureus; and Gram-negative: Citrobacter freundii, Escherichia coli, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella enterica, Serratia marcescens and Vibrio harveyi) to various flavonoids: genistein and daidzein (isoflavones), apigenin (a flavone), naringenin (a flavanone) and kaempferol (a flavonol) by measurement of generation times of bacteria in liquid cultures. The presented results indicate that this simple method is adequate for unambiguous assessment of sensitivity of bacterial strains to flavonoids.     

Development and validation of a real-time quantitative PCR assay for rapid identification of Bacillus anthracis in environmental samples

A real-time polymerase chain reaction (PCR) assay was developed for rapid identification of Bacillus anthracis in environmental samples. These samples often harbor Bacillus cereus bacteria closely related to B. anthracis, which may hinder its specific identification by resulting in false positive signals. The assay consists of two duplex real-time PCR: the first PCR allows amplification of a sequence specific of the B. cereus group (B. anthracis, B. cereus, Bacillus thuringiensis, Bacillus weihenstephanensis, Bacillus pseudomycoides, and Bacillus mycoides) within the phosphoenolpyruvate/sugar phosphotransferase system I gene and a B. anthracis specific single nucleotide polymorphism within the adenylosuccinate synthetase gene. The second real-time PCR assay targets the lethal factor gene from virulence plasmid pXO1 and the capsule synthesis gene from virulence plasmid pXO2. Specificity of the assay is enhanced by the use of minor groove binding probes and/or locked nucleic acids probes. The assay was validated on 304 bacterial strains including 37 B. anthracis, 67 B. cereus group, 54 strains of non-cereus group Bacillus, and 146 Gram-positive and Gram-negative bacteria strains. The assay was performed on various environmental samples spiked with B. anthracis or B. cereus spores. The assay allowed an accurate identification of B. anthracis in environmental samples. This study provides a rapid and reliable method for improving rapid identification of B. anthracis in field operational conditions.     

Effects of 5-fluoro-2′-deoxyuridine on bacterial growth its use for DNA labelling

Experiments on the action of 5-fluoro-2′-deoxyuridine on growth ofEscherichia coli B, CECT 101;Pseudomonas fluorescens, CECT 318;Pseudomonas savastanoi, CECT 93;Micrococcus luteus, ATCC 4698;Bacillus cereus, CIP 52.58;Bacillus macerans, ClP 52.58 andBacillus subtilis, ATCC 6633, are described. The inhibition of growth is reversed by thymine plus uracil in all cases except inPseudomonas strains in which uracil alone is active, and in which no exogenous thymine is taken up, not even in the presece of 2′-deoxyguanosine. Growth conditions for improved labelling of bacterial DNA are discussed in the light of the results.