Simultaneous removal of crude oil and heavy metals by highly adapted bacterial strain Cutibacterium sp. NL2 isolated from Algerian oilfield

Investigating the ability of bacteria to simultaneously enhance hydrocarbon removal and reduce heavy metals’ toxicity is necessary to design more effective bioremediation strategies. A bacterium (NL2 strain) isolated from an Algerian oilfield was cultivated on crude oil as sole carbon and energy sources. Molecular analyses of the 16S rRNA gene sequence placed the strain within the Cutibacterium genera. This isolate was able to tolerate up to 60% of crude oil as sole carbon source. Chemical analyses (GC-MS) evidenced that strain NL2 was able to degrade 92.22% of crude oil (at optimal growing conditions: pH 10, 44 °C, 50 g L−1 NaCl, and 20% of crude oil (v/v) as sole carbon source) in only 7 days. NL2 isolate was also able to produce biosurfactants with reduction of surface tension of growing media (29.4 mN m−1). On the other hand, NL2 strain was able to tolerate high lead (Pb) and copper (Cu) concentrations (up to 60 mM). In fact, NL2 cultivated in the presence of 20% of crude oil, and 0.48 mM of Pb was able to reduce Pb concentration by a 41.36%. In turn, when cultivated on high Pb concentration (15 mM), the strain was able to remove 35.19% of it and 86.25% of crude oil, both in a time frame of 7 days. Our findings suggest that Cutibacterium strain NL2 is able to efficiently use and remove a wide range of crude oil substrates in presence of high Pb concentration. Accordingly, NL2 strain is of extreme interest from a biotechnological standpoint. (AU)

Evaluation of the engineering properties of asphaltic concrete composite produced from recycled asphalt pavement and polyethylene plastic.

This study investigated the suitability of recycled asphalt pavement and polyethylene wastes as coarse aggregate in asphaltic concrete by evaluating the impact of the use of polyethylene polymer wastes and recycled asphalt pavement composite as aggregates on the physical and mechanical properties of the asphaltic concrete. The physical characteristics of the aggregate and bitumen were determined using relevant parametric tests. Recycled asphalt pavement was used to make asphaltic concrete samples using LDPE at 5%, 10%, 15%, RAP at 5% and HDPE at 5%, 10%, 15%, and a mixture of LDPE + HDPE at 5+5%, 7.5+7.5% and 10+10% RAP at 5% as additives. Marshall Stability test was conducted to assess the mechanical strength of the asphaltic concrete, and the results included information on the aggregate's stability, flow, density, voids filled with bitumen, voids filled with air, and voids in mineral aggregate. In addition, the surface and crystal structure of the aggregates was studied by carrying out a microscopic examination with a Scanning Electron Microscope (SEM) and X-Ray diffraction (XRD). The results obtained from this study demonstrated that RAP, HDPE & LDPE are viable conventional aggregate substitute for asphalt concrete production.

Hydrocarbons in Formicidae: influence of chemical footprints on ant behavioral strategies.

When an insect walks, it leaves chemical cues that derive from the arolium, a tarsal structure. These cues may contain important information about other species that occur in their community and can then mediate interactions of competition, predation, and information about resources with ants from their own colony. The compounds of these cues are released into the substrate in the form of chemical footprints. There are still few species studied, and little is known about the behavior of ants regarding these signals and how they use them in their interactions. Therefore, the aim of this study was to assess the behavioral strategy of different ant species when confronted with chemical footprints left by other ants, as well as identify their compounds and their relationship with the cuticular hydrocarbon profile. The experiments were performed using a Y-maze, where in one of the arms, there were chemical footprints of their own species or of other species, and the other Y arm was footprint-free. The chemical compounds of footprints and cuticle were analyzed by gas chromatography-mass spectrometry. The results show that foragers of all species detect and respond to the presence of chemical cues in the form of footprints left by other ants. Foragers of all species followed footprints of individuals of the same species both nestmates and non-nestmates; however, Neoponera villosa avoided the footprints of Cephalotes borgmeieri, and C. borgmeieri avoided the footprints of the other two species. The chemical compositions of the cuticle and footprints are related to each other and are specific to each species.

Experimental study on the effect of Coumarone resin on the performance of SBS-modified asphalt.

An inexpensive and high-performing solid Coumarone resin was added to Styrene-butadiene-styrene (SBS) copolymer-modified asphalt to enhance its storage stability and road performance. To assess the effect of Coumarone resin dosage on the SBS-modified asphalt, a series of laboratory tests were conducted. The composite modified asphalt's segregation test was used to evaluate its storage stability, Dynamic Shear Rheometer (DSR) and Multiple Stress Creep Recovery (MSCR) tests were employed to investigate its high-temperature performance and permanent deformation resistance, and the Bending Beam Rheology (BBR) test was utilized to measure its low-temperature performance. Fluorescence microscopy was used to observe the composite modified asphalt's microstructure, and Fourier Transform Infrared Spectroscopy (FTIR) was conducted to study the changes in chemical structure during the modification process. The results showed that Coumarone resin can improve the compatibility of SBS and asphalt, improve the high-temperature performance and deformation resistance of SBS-modified asphalt, and adding an appropriate amount of Coumarone resin can help enhance the low-temperature cracking resistance of modified asphalt. The optimal dosage of Coumarone resin recommended for SBS-modified asphalt performance enhancement is 2% under the test conditions, as determined by comparing the test results of samples with various dosages.

Hydrocarbon stapled temporin-L analogue as potential antibacterial and antiendotoxin agents with enhanced protease stability.

Antimicrobial resistance (AMR) is a serious global concern and a huge burden on the healthcare system. Antimicrobial peptides (AMPs) are considered as a solution of AMR due to their membrane-lytic and intracellular mode of action and therefore resistance development against AMPs is less frequent. One such AMPs, temporin-L (TL) is a 13-mer peptide reported as a potent and broad-spectrum antibacterial agent with significant immunomodulatory activity. However, TL is toxic to human erythrocytes at their antibacterial concentrations and therefore various analogues were synthesized with potent antimicrobial activity and lower hemolytic activity. In this work, we have selected a non-toxic engineered analogue of TL (eTL) and performed hydrocarbon stapling of amino acid residues at i to i + 4 positions at different part of sequence. The synthesized peptides were investigated against both the gram-positive and gram-negative bacteria as well as methicillin resistant S. aureus, its MIC was measured in the concentrations range of 0.9-15.2 µM. All analogues were found equal or better antibacterial as compared to parent peptide. Interestingly one analogue eTL [5-9] was found to be non-cytotoxic and stable in presence of the human serum. Mode of action studies revealed membrane depolarizing and disruptive mode of action with live MRSA. Further in vivo studies of antimicrobial against MRSA infection and anti-endotoxin activities in mice model revealed potential activity of the stapled peptide analogue. Overall, this reports on stapled analogue of the AMPs highlights an important strategy for the development of new antibacterial therapeutics against AMR.

Release Pattern of Light Aromatic Hydrocarbons during the Biomass Roasting Process.

Roasting is an important step in the pretreatment of biomass upgrading. Roasting can improve the fuel quality of biomass, reduce the O/C and H/C ratios in the biomass, and provide the biomass with a fuel quality comparable to that of lignite. Therefore, studying the structure and component evolution laws during biomass roasting treatment is important for the rational and efficient utilization of biomass. When the roasting temperature is 200-300 °C, the cellulose and hemicellulose in the biomass undergo a depolymerization reaction, releasing many monocyclic aromatic hydrocarbons with high reactivity. The proportion of monocyclic aromatic hydrocarbons in biomass roasting products can be effectively regulated by controlling the reaction temperature, residence time, catalyst, baking atmosphere, and other factors in the biomass roasting process. This paper focuses on the dissociation law of organic components in the pretreatment process of biomass roasting.

Broad-spectrum hydrocarbon-degrading microbes in the global ocean metagenomes.

Understanding the diversity and functions of hydrocarbon-degrading microorganisms in marine environments is crucial for both advancing knowledge of biogeochemical processes and improving bioremediation methods. In this study, we leveraged nearly 20,000 metagenome-assembled genomes (MAGs), recovered from a wide array of marine samples across the global oceans, to map the diversity of aerobic hydrocarbon-degrading microorganisms. A broad bacterial diversity was uncovered, with a notable preference for degrading aliphatic hydrocarbons over aromatic ones, primarily within Proteobacteria and Actinobacteriota. Three types of broad-spectrum hydrocarbon-degrading bacteria were identified for their ability to degrade various hydrocarbons and possession of multiple copies of hydrocarbon biodegradation genes. These bacteria demonstrate extensive metabolic versatility, aiding their survival and adaptability in diverse environmental conditions. Evidence of gene duplication and horizontal gene transfer in these microbes suggested a potential enhancement in the diversity of hydrocarbon-degrading bacteria. Positive correlations were observed between the abundances of hydrocarbon-degrading genes and environmental parameters such as temperature (-5 to 35 °C) and salinity (20 to 42 PSU). Overall, our findings offer valuable insights into marine hydrocarbon-degrading microorganisms and suggest considerations for selecting microbial strains for oil pollution remediation.

Biosurfactants production by marine yeasts isolated from zoanthids and characterization of an emulsifier produced by Yarrowia lipolytica LMS 24B.

The present study investigates the potential for biosurfactant production of 19 marine yeast species obtained from zoanthids. Using the emulsification index test to screen the samples produced by the marine yeasts, we verified that five isolates exhibited an emulsification index ≥50%. Additional tests were performed on such isolates, including oil displacement, drop collapse, Parafilm M assay, and surface tension measurement. The tolerance of produced biosurfactants for environmental conditions was also analyzed, especially considering the media's temperature, pH, and salinity. Moreover, the surfactant's ability to emulsify different hydrocarbon sources and to metabolize kerosene as the sole carbon source was evaluated in vitro. Our results demonstrate that yeast biosurfactants can emulsify hydrocarbon sources under different physicochemical conditions and metabolize kerosene as a carbon source. Considering the Yarrowia lipolytica LMS 24B as the yeast model for biosurfactant production from the cell's wall biomass, emulsification indexes of 61.2% were obtained, even at a high temperature of 120 °C. Furthermore, the Fourier-transform middle infrared spectroscopy (FTIR) analysis of the biosurfactant's chemical composition revealed the presence of distinct functional groups assigned to a glycoprotein complex. Considering the status of developing new bioproducts and bioprocesses nowadays, our findings bring a new perspective to biosurfactant production by marine yeasts, especially Y. lipolytica LMS 24B. In particular, the presented results validate the relevance of marine environments as valuable sources of genetic resources, i.e., yeast strains capable of metabolizing and emulsifying petroleum derivatives.

Association between Ambient Volatile Organic Compounds Exposome and Emergency Hospital Admissions for Cardiovascular Disease.

The limited research on volatile organic compounds (VOCs) has not taken into account the interactions between constituents. We used the weighted quantile sum (WQS) model and generalized linear model (GLM) to quantify the joint effects of ambient VOCs exposome and identify the substances that play key roles. For a 0 day lag, a quartile increase of WQS index for n-alkanes, iso/anti-alkanes, aromatic, halogenated aromatic hydrocarbons, halogenated saturated chain hydrocarbons, and halogenated unsaturated chain hydrocarbons were associated with 1.09% (95% CI: 0.13, 2.06%), 0.98% (95% CI: 0.22, 1.74%), 0.92% (95% CI: 0.14, 1.69%), 1.03% (95% CI: 0.14, 1.93%), 1.69% (95% CI: 0.48, 2.91%), and 1.85% (95% CI: 0.93, 2.79%) increase in cardiovascular disease (CVD) emergency hospital admissions, respectively. Independent effects of key substances on CVD-related emergency hospital admissions were also reported. In particular, an interquartile range increase in 1,1,1-trichloroethane, methylene chloride, styrene, and methylcyclohexane is associated with a greater risk of CVD-associated emergency hospital admissions [3.30% (95% CI: 1.93, 4.69%), 3.84% (95% CI: 1.21, 6.53%), 5.62% (95% CI: 1.35, 10.06%), 8.68% (95% CI: 3.74, 13.86%), respectively]. We found that even if ambient VOCs are present at a considerably low concentration, they can cause cardiovascular damage. This should prompt governments to establish and improve concentration standards for VOCs and their sources. At the same time, policies should be introduced to limit VOCs emission to protect public health.

Rheological properties of warm mixed high viscosity asphalt at high and low temperatures.

The rheological properties of asphalt can well reflect its road performance, but the rheological properties of warm mix high viscosity asphalt (HVA) are unclear. In order to study the effect of warm mixing agent on rheological properties of HVA, two kinds of warm mixing agent EC120 (EC) and Evotherm M1 (M1) were selected to prepare warm mix HVA. The rheological properties of warm mix HVA at high temperature (135~195°C), medium temperature (0~80°C) and low temperature (-6~18°C) were studied by Brinell rotary viscosity test, dynamic shear rheological test (including temperature scanning, frequency scanning, linear amplitude scanning) and bending beam rheological test. The test results show that both EC and M1 have good viscosity reduction effect on HVA at high temperature, and can effectively reduce the construction temperature. At medium temperature, M1 can effectively improve the fatigue resistance of HVA, and the fatigue life can be increased by about 30% when the dosage is 0.6%. EC can increase the rutting factor of HVA and improve its resistance to deformation, but it will reduce its fatigue performance. When the dosage is 4%, the fatigue life will be reduced by about 9%. At low temperature, M1 can reduce the creep stiffness S, increase the creep rate m, and improve the low temperature performance of HVA, while EC has the opposite effect, weakening the low temperature performance of HVA. The results are helpful to understand the rheological properties of warm mix HVA and promote its application.

The interactive force between fat, oil, and grease (FOG) deposits and the effect of food waste disposers on kitchen drainage systems.

Food waste disposers (FWDs) streamline kitchen waste management and facilitate waste classification, whether they would increase the potential of blockage in kitchen drainage system is still unknown. This study conducted a theoretical analysis of the interactive forces between fat, oil, and grease (FOG) deposits and their aggregation on pipe walls. The study involved grading food waste particles processed by FWDs using sieving and weighing techniques to determine the mean weight diameter (MWD) of various aggregations. A full-scale experimental system, implemented in a 60-m high test tower, simulated blockages in horizontal pipes of high-rise buildings. The effect of pipeline materials and particle sizes on blockage were examined by measuring the adhesion of deposits on horizontal pipes. Energy dispersive spectrometer (EDS) analysis suggested that liquid bridge force is a primary factor in aggregate formation. Hand-cut particles formed aggregates with the highest MWD value. Particle size analysis revealed that sizes ranging from 2.36 to 4.75 mm, 1.18-2.36 mm, and 0.60-1.18 mm constituted over 80 % of particles ground by FWDs, with an average size of 2.16 mm. Results of full-scale experiment indicate particle diameters, friction coefficients and lipophilic coefficient significantly affected the propensity of these aggregates to adhere to pipes. Notably, particles processed by FWDs tended to cause blockages more frequently than hand-cut particles. These findings elucidate the deposition mechanism of FOG deposits and offer strategies to reduce blockages in kitchen drainage systems, such as reducing current grinding particle size by 18 % to 1.77 mm or selecting pipes like cast iron and high-density polyethylene.

Evaluation of biomass sources on the production of biofuels from lignocellulosic waste over zeolite catalysts.

Catalytic fast pyrolysis (CFP) is a promising method to convert biomass waste into sustainable bio-oils. However, the relationship gap between biomass characteristics and bio-oil quality has hindered the development of CFP technology. This study investigated the pyrolysis and CFP of ten biomass sources over zeolites, and showed that biomass sources and zeolites played important roles in bio-oil production. For noncatalytic trials, the bio-oil yield was positively related to holocellulose (R2 = 0.75) and volatiles content (R2 = 0.62) but negatively to ash content (R2 = -0.65). The bio-oil quality was dramatically improved after catalyst addition. For CFP over ZSM-5, hydrocarbons selectivity of bio-oils was increased by 1.6∼79.3 times, which was closely related to H/C ratio (R2 = 0.79). For ZSM-5@SBA-15 trials, the dependency of hydrocarbons selectivity on biomass characteristics was less clear than that in ZSM-5 counterparts, although undesirable PAHs were inhibited for most biomass sources. This study demonstrated the influence mechanism of biomass characteristics on bio-oil compositions.

Research on surface treatment technology for quickly improving the skid resistance of tunnel concrete pavement.

In order to solve the problem that the skid resistance of concrete pavement in tunnel deteriorates rapidly, which is easy to cause traffic accidents, the anti-skid rapid elevation technology of surface treatment is proposed. Wear tests were used to investigate the effects of concrete surface roughness, properties of modified emulsified asphalt binder and anti-skid fine aggregate type on long-term skid resistance of treated surfaces. The results show that the four coarsening methods of fine milling, milling, grooving and brooming can improve the skid resistance of concrete, and the skid resistance durability of fine milling and milling is better. The adhesive property of modified emulsified asphalt is the best when the content of water-based epoxy resin is 20%. In different aggregates, the anti-skid effect is better when silicon carbide is used as anti-skid aggregate and the particle size is 0.6mm:0.3mm = 2:3. The method of fine milling of concrete surface + spraying epoxy emulsified asphalt + spreading silicon carbide can effectively improve the anti-skid performance of the original concrete pavement, and the feasibility of the scheme is verified by the test road. The research results have a good reference value for improving the skid resistance of tunnel concrete pavement.

Bioremediation of oily hypersaline soil via autochthonous bioaugmentation with halophilic bacteria and archaea.

Kuwaiti hypersaline soil samples were contaminated with 5 % (w/w) weathered Kuwaiti light crude oil and bioaugmented with autochthonous halophilic hydrocarbonoclastic archaeal and bacterial strains, two each, individually and as consortia. Residual oil contents were determined, and microbial communities were analyzed by culture-dependent and culture-independent approaches initially and seasonally for one year. After one year of the bioremediation process, the mean oil degradation rate was similar across all treated soils including the controlled unbioaugmented one. Oil hydrocarbons were drastically reduced in all soil samples with values ranging from 82.7 % to 93 %. During the bioremediation process, the number of culturable oil-degrading bacteria increased to a range of 142 to 344 CFUx104 g-1 after 12 months of bioaugmentation. Although culture-independent analysis showed a high proportion of inoculants initially, none could be cultured throughout the bioremediation procedure. Within a year, microbial communities changed continually, and 33 species of halotolerant/halophilic hydrocarbonoclastic bacteria were isolated and identified belonged mainly to the three major bacterial phyla Actinobacteria, Proteobacteria, and Firmicutes. The archaeal phylum Halobacterota represented <1 % of the microbial community's relative abundance, which explains why none of its members were cultured. Improving the biodegradability of an already balanced environment by autochthonous bioaugmentation is more involved than just adding the proper oil degraders. This study emphasizes the possibility of a relatively large resistant population, a greater diversity of oil-degrading microorganisms, and the highly selective impacts of oil contamination on hypersaline soil bacterial communities.

Unlocking bioremediation potential for site restoration: A comprehensive approach for crude oil degradation in agricultural soil and phytotoxicity assessment.

Crude oil contamination has inflicted severe damage to soil ecosystems, necessitating effective remediation strategies. This study aimed to compare the efficacy of four different techniques (biostimulation, bioaugmentation, bioaugmentation + biostimulation, and natural attenuation) for remediating agricultural soil contaminated with crude oil using soil microcosms. A consortium of previously characterized bacteria Xanthomonas boreopolis, Microbacterium schleiferi, Pseudomonas aeruginosa, and Bacillus velezensis was constructed for bioaugmentation. The microbial count for the constructed consortium was recorded as 2.04 ± 0.11 × 108 CFU/g on 60 d in augmented and stimulated soil samples revealing their potential to thrive in chemically contaminated-stress conditions. The microbial consortium through bioaugmentation + biostimulation approach resulted in 79 ± 0.92% degradation of the total polyaromatic hydrocarbons (2 and 3 rings âˆ¼ 74%, 4 and 5 rings âˆ¼ 83% loss) whereas, 91 ± 0.56% degradation of total aliphatic hydrocarbons (C8-C16 ∼ 90%, C18-C28 ∼ 92%, C30 to C40 ∼ 88% loss) was observed in 60 d. Further, after 60 d of microcosm treatment, the treated soil samples were used for phytotoxicity assessment using wheat (Triticum aestivum), black chickpea (Cicer arietinum), and mustard (Brassica juncea). The germination rates for wheat (90%), black chickpea (100%), and mustard (100%) were observed in 7 d with improved shoot-root length and biomass in both bioaugmentation and biostimulation approaches. This study projects a comprehensive approach integrating bacterial consortium and nutrient augmentation strategies and underscores the vital role of innovative environmental management practices in fostering sustainable remediation of oil-contaminated soil ecosystems. The formulated bacterial consortium with a nutrient augmentation strategy can be utilized to restore agricultural lands towards reduced phytotoxicity and improved plant growth.

Study on crack resistance of self-healing microcapsules in asphalt pavement by multi-scale method.

Self-healing microcapsules in the asphalt pavement must be kept intact under vehicle load to ensure there is enough rejuvenator in capsules when cracks appear in asphalt pavement. In this paper, the crack resistance of self-healing microcapsules in asphalt pavement was evaluated. Firstly, an expanding multi-scale analysis was conducted based on proposed mesoscopic mechanical models with the aim to determine the mechanical parameters for the following contracting multi-scale analysis. Secondly, the periodic boundary condition was introduced for the contracting multi-scale analysis and the stress field of the capsule wall was obtained. Finally, the effects of the design parameters of the microcapsule on its crack resistance in asphalt pavement were investigated. The results showed that the incorporation of microcapsules has almost no effect on the elastic constants of the asphalt mixture. The core could be simplified as an approximately incompressible solid with the elastic constants determined by the proposed mesoscopic mechanical model. With the increase of the modulus of the capsule wall, the mean maximum tensile stress of the capsule wall increased from 0.372 MPa to 0.465 MPa, while with the decrease of the relative radius of the capsule core, the mean maximum tensile stress of the capsule wall increased from 0.349 MPa to 0.461 MPa. The change in the mean maximum tensile stress of the capsule wall caused by the change of capsule diameter was within 5%. The relative radius of the capsule core and the elastic modulus of capsule wall were two key parameters in capsule design. Besides, the microcapsules with the wall made of resin would not crack under the vehicle load before microcracks occurred in asphalt pavement.

Rhinoceros beetle (Trypoxylus dichotomus) cuticular hydrocarbons contain information about body size and sex.

Japanese rhinoceros beetle (Trypoxylus dichotomus) males have exaggerated horns that are used to compete for territories. Larger males with larger horns tend to win these competitions, giving them access to females. Agonistic interactions include what appears to be assessment and often end without escalating to physical combat. However, it is unknown what information competitors use to assess each other. In many insect species chemical signals can carry a range of information, including social position, nutritional state, morphology, and sex. Specifically, cuticular hydrocarbons (CHCs), which are waxes excreted on the surface of insect exoskeletons, can communicate a variety of information. Here, we asked whether CHCs in rhinoceros beetles carry information about sex, body size, and condition that could be used by males during assessment behavior. Multivariate analysis of hydrocarbon composition revealed patterns associated with both sex and body size. We suggest that Rhinoceros beetles could be communicating information through CHCs that would explain behavioral decisions.

Leaching behavior of metals from asphalt mixtures modified with crumb rubber from scrap tires.

There are concerns about the potential toxicity of bitumen and recycled materials such as reclaimed asphalt pavements from end-of-life roads and crumb rubber from scrap tires used in asphalt mixtures because they contain metals that may be released into the groundwater. This study investigated the potential metal leaching of laboratory-prepared asphalt mixtures modified with polymer coated rubber (PCR) with wet and dry technology, devulcanized rubber (DVR), compared to an unmodified control mixture and a blend modified with a synthetic polymer (SBS). The objectives were to i) quantify concentrations of metals released, ii) calculate the flux rate, the cumulative mass release, and the assessment ratio for each metal, iii) verify if the metals exceeded the EPA drinking water limit, and, finally, iv) assess the source of metals release. Zinc had the highest concentration among all metals and was present in eluates from all mixtures. The cumulative zinc concentration from DVR mixture was 41% and 34% higher than the control and SBS mixtures, respectively. For PCR wet, the cumulative zinc concentration was 9% higher than the control blend and 1% lower than the SBS mix. The assessment ratio indicated that all metal concentrations would not exceed the drinking water limit, except for zinc, for which further evaluations were required. The main source of zinc may derive from aggregates. This work showed that crumb rubber might not be the only source of metal leaching, and its use in asphalt pavements does not cause a metal leaching higher than other materials.

Molecular and metabolic characterization of petroleum hydrocarbons degrading Bacillus cereus.

Hydrocarbon constituents of petroleum are persistent, bioaccumulated, and bio-magnified in living tissues, transported to longer distances, and exert hazardous effects on human health and the ecosystem. Bioaugmentation with microorganisms like bacteria is an emerging approach that can mitigate the toxins from environmental sources. The present study was initiated to target the petroleum-contaminated soil of gasoline stations situated in Lahore. Petroleum degrading bacteria were isolated by serial dilution method followed by growth analysis, biochemical and molecular characterization, removal efficiency estimation, metabolites extraction, and GC-MS of the metabolites. Molecular analysis identified the bacterium as Bacillus cereus, which exhibited maximum growth at 72 hours and removed 75% petroleum. Biochemical characterization via the Remel RapID™ ONE panel system showed positive results for arginine dehydrolase (ADH), ornithine decarboxylase (ODC), lysine decarboxylase (LDC), o-nitrophenyl-ß-D-galactosidase (ONPG), p-nitrophenyl-ß-D-glucosidase (ßGLU), p-nitrophenyl-N-acetyl-ß-D-glucosaminidase (NAG), malonate (MAL), adonitol fermentation (ADON), and tryptophane utilization (IND). GC-MS-based metabolic profiling identified alcohols (methyl alcohol, o-, p- and m-cresols, catechol, and 3-methyl catechol), aldehydes (methanone, acetaldehyde, and m-tolualdehyde), carboxylic acid (methanoic acid, cis,cis-muconic acid, cyclohexane carboxylic acid and benzoic acid), conjugate bases of carboxylic acids (benzoate, cis,cis-muconate, 4-hydroxybenzoate, and pyruvate) and cycloalkane (cyclohexene). It suggested the presence of methane, methylcyclohexane, toluene, xylene, and benzene degradation pathways in B. cereus.

Biomass and hydrocarbon production from Botryococcus braunii: A review focusing on cultivation methods.

Botryococcus braunii has garnered significant attention in recent years due to its ability to produce high amounts of renewable hydrocarbons through photosynthesis. As the world shifts towards a greener future and seeks alternative sources of energy, the cultivation of B. braunii and the extraction of its hydrocarbons can potentially provide a viable solution. However, the development of a sustainable and cost-effective process for cultivating B. braunii is not without challenges. Compared to other microalgae, B. braunii grows very slowly, making it time-consuming and expensive to produce biomass. In response to these challenges, several efforts have been put into optimizing Botryococcus braunii cultivation systems to increase biomass growth and hydrocarbon production efficiency. This review presents a comparative analysis of different Botryococcus braunii cultivation systems, and the factors affecting the productivity of biomass and hydrocarbon in Botryococcus braunii are critically discussed. Attached microalgal growth offers several advantages that hold significant potential for enhancing the economic viability of microalgal fuels. Here, we propose that employing attached growth cultivation, coupled with the milking technique for hydrocarbon extraction, represents an efficient approach for generating renewable fuels from B. braunii. Nevertheless, further research is needed to ascertain the viability of large-scale implementation.