Bacteria as a source of biopigments and their potential applications.

From the prehistoric period, the utilization of pigments as colouring agents was an integral part of human life. Early people may have utilized paint for aesthetic motives, according to archaeologists. The pigments are either naturally derived or synthesized in the laboratory. Different studies reported that certain synthetic colouring compounds were toxic and had adverse health and environmental effects. Therefore, knowing the drawbacks of these synthetic colouring agents now scientists are attracted towards the harmless natural pigments. The main sources of natural pigments are plants, animals or microorganisms. Out of these natural pigments, microorganisms are the most important source for the production and application of bioactive secondary metabolites. Among all kinds of microorganisms, bacteria have specific benefits due to their short life cycle, low sensitivity to seasonal and climatic variations, ease of scaling, and ability to create pigments of various colours. Based on these physical characteristics, bacterial pigments appear to be a promising sector for novel biotechnological applications, ranging from functional food production to the development of new pharmaceuticals and biomedical therapies. This review summarizes the need for bacterial pigments, biosynthetic pathways of carotenoids and different applications of bacterial pigments.

A biosurfactant-producing novel bacterial strain isolated from vermicompost having multiple plant growth-promoting traits.

The nutrient-rich vermicompost which is used as manure for the growth and development of plants is rich in microbial flora. These microbes protect the plants against several infectious pathogenic microbes. As certain microbes are known to produce biosurfactants as metabolites, an investigation was carried out to isolate biosurfactant-producing bacterial strains from vermicompost with the efficient antifungal property. From the study, it was revealed that biosurfactant-producing bacterial strains are present in the vermicompost. A total of nine bacterial strains were isolated from the vermicompost. Among them, one most efficient biosurfactant-producing bacterial strains with antifungal properties have been screened. After molecular characterization of the isolated strain, it was revealed that the bacterial strain is Bacillus licheniformis strain SCV1. The strain produces 3.4 ± 0.1 g/L of crude biosurfactant, which when column purified yields 3.1 ± 0.1 g/L of biosurfactant. The biosurfactant exhibited excellent emulsifying activity (E24 ) of 96.56% against crude oil. The produced biosurfactant was identified as a lipopeptide consisting of a mixer of surfactin and iturin. Furthermore, the biosurfactant exhibited significant antifungal activity against a wide range of phytopathogens, showing 76.3% inhibition against Sclerotinia sclerotiorum, 53% inhibition against Colletotrichum gloeosporioides, 51% against Fusarium verticillioides, and 36% against Corynespora cassicolla. Along with antifungal activities, the stain was found to exhibit multiple plant growth-promoting traits. This study, thus indicates that vermicompost might contain biosurfactant-producing microbes which can render protection to the plant against various phytopathogens by the production of biosurfactants and can also stimulate plant growth.

Soil treatment using a biosurfactant producing bacterial consortium in rice fields contaminated with oily sludge- a sustainable approach.

A consortium of two biosurfactant-producing bacteria (Bacillus pumilus KS2 and Bacillus cereus R2) was developed to remediate petroleum hydrocarbon-contaminated paddy soil. Soil samples from a heavily contaminated rice field near Assam's Lakwa oilfield were collected and placed in earthen pots for treatment. After each month of incubation, 50 g of soil from each earthen pot was collected, and the soil TPH (ppm) in each sample was determined. The extracted TPH samples were analysed by Gas chromatography-mass spectrometry (GC-MS) to confirm microbial degradation. The soil samples were examined for changes in pH, conductivity, total organic content (TOC), water holding capacity, and total nitrogen content in addition to TPH degradation. An increasing trend in TPH degradation was observed with each passing month. After six months of treatment, the sample with the lowest initial TPH concentration (1735 ppm) had the highest degradation (91.24%), while the soil with the highest amount of TPH (5780 ppm) had the lowest degradation (74.35%). A wide range of aliphatic hydrocarbons found in soil samples was degraded by the bacterial consortium. The soil samples contained eight different low- and high-molecular-weight PAHs. Some were fully mineralized, while others were significantly reduced. With the decrease in the TPH level in the polluted soil, a significant improvement in the soil's physicochemical qualities (such as pH, electrical conductivity, total organic content, and water-holding capacity) was observed.

Wound healing efficacy of rhamnolipid-coated zinc oxide nanoparticle along with its in vivo antibacterial efficacy against Staphylococcus aureus.

Rhamnolipids are microbial metabolites with antibacterial efficacies, which can be further boosted through the application of nanobiotechnology. In this study, the efficacy of rhamnolipid-coated zinc oxide nanoparticles (ZnRL) has been studied for their wound healing efficacy as well as in vivo antibacterial efficacy. Thus, this study evaluates the efficacy of ZnRL to heal an excised infected wound, which was compared with the healing efficacy of rhamnolipid and clindamycin. The study revealed that rhamnolipid-coated zinc oxide nanoparticles possess promising wound healing efficacy with prominent antibacterial activity in the rat model. Prominent wound healing in a Staphylococcus aureus infected excised wound was observed on the 5th day of the treatment when the wound site was treated with 100 µl of 0.5 mg/ml of ZnRL. This concentration of ZnRL was found to exhibit efficient antibacterial activity against the pathogen, thereby decreasing the amount of pathogen in the wound site. ZnRL exhibited efficient wound contraction, thereby decreasing the size of the wound prominently in 5 days. Histological study revealed efficient tissue remodelling in ZnRL-treated skin which resulted in rapid formation of the epidermis and recruitment of various dermal cells within the 5th day of treatment. The study also revealed the non-cytotoxic effect of the nanoparticles in fibroblast cell line L929 and the non-haemolytic effect against blood cells, indicating its potential in pharmaceuticals.

Biodegradation of hazardous naphthalene and cleaner production of rhamnolipids - Green approaches of pollution mitigation.

Toxic and hazardous waste poses a serious threat to human health and the environment. Green remediation technologies are required to manage such waste materials, which is a demanding and difficult task. Here, effort was made to explore the role of Pseudomonas aeruginosa SR17 in alleviating naphthalene via catabolism and simultaneously producing biosurfactant. The results showed up to 89.2% naphthalene degradation at 35 °C and pH 7. The GC/MS analysis revealed the generation of naphthalene degradation intermediates. Biosurfactant production led to the reduction of surface tension of the culture medium to 34.5 mN/m. The biosurfactant was further characterized as rhamnolipids. LC-MS of the column purified biosurfactant revealed the presence of both mono and di rhamnolipid congeners. Rhamnolipid find tremendous application in medical field and as well as in detergent industry and since they are of biological origin, they can be used as favorable alternative against their chemical counterparts. The study demonstrated that catabolism of naphthalene and concurrent formation of rhamnolipid can result in a dual activity process, namely environmental cleanup and production of a valuable microbial metabolite. Additionally, the present-day application of rhamnolipids is highlighted.

Synthesis, characterization, and evaluation of antibacterial efficacy of rhamnolipid-coated zinc oxide nanoparticles against Staphylococcus aureus.

The antimicrobial efficacy of rhamnolipid is well established against a wide range of pathogens. However little is known about the enhancement of antimicrobial efficacy of rhamnolipid in the form of nanoparticles. With a curiosity of enhancing antimicrobial activity, a study has been carried out to evaluate the antimicrobial efficacy of rhamnolipid-coated zinc oxide nanoparticles. The zinc oxide nanoparticles were synthesized with rhamnolipid, produced by Pseudomonas aeruginosa JS29. The rhamnolipid-coated zinc oxide nanoparticles were characterized by FTIR, XRD, TGA, TEM, and SAED. The antimicrobial and antibiofilm efficacy of the nanoparticles was evaluated against Staphylococcus aureus MTCC 96. FTIR, XRD, TEM, and SAED analyses confirmed that the nanoparticles contain both rhamnolipid and zinc as constituents and are polycrystalline with sizes ranging from 40 to 50 nm. At a concentration of 250 µg/ml, rhamnolipid-coated zinc oxide nanoparticles exhibited 80% growth inhibition of the pathogen. Again, at the same concentration, the nanoparticle was observed to inhibit 78% of biofilm formation while disrupting 100% of preformed biofilm. The nanoparticles demonstrated an enhanced inhibitory and antibiofilm efficacy against the pathogen compared to the individual effect of both rhamnolipid and zinc oxide nanoparticles. With the established non-toxicity of rhamnolipid-coated zinc oxide nanoparticles in fibroblast cell lines, the nanoparticles could be a promising pharmaceutical alternative.

Rhamnolipid exhibits anti-biofilm activity against the dermatophytic fungi Trichophyton rubrum and Trichophyton mentagrophytes.

Dermatophytes are responsible for a majority of fungal infections in humans and other vertebrates, causing dermatophytosis. Treatment failures are often associated with biofilm formation, making dermatophytes resistant to antifungals. In this study, effects of a rhamnolipid (RL-SS14) produced by Pseudomonas aeruginosa SS14 on planktonic cells of Trichophyton rubrum and Trichophyton mentagrophytes, their biofilm formation, and disruption of mature biofilms were assessed. The composition of RL-SS14 was analysed using FTIR, HPLC-ESI-MS, and GC-MS. Minimum inhibitory concentrations against the planktonic forms of T. rubrum and T. mentagrophytes were 0.5 mg/mL and 0.125 mg/mL, respectively. Crystal-violet (biofilm biomass) and safranin (extracellular matrix) staining revealed that RL-SS14 significantly inhibited biofilm formation and also reduced preformed biofilms in a dose-dependent manner. Microscopic visualization of treated biofilms via SEM, AFM, and CLSM revealed marked morphological damage, cell death, and reduced extracellular matrix. The results indicate the potential of RL-SS14 as an anti-biofilm agent against dermatophytes.

Isolation of a novel rhizobacteria having multiple plant growth promoting traits and antifungal activity against certain phytopathogens.

The present study was conducted to explore the potentialities of a versatile multifarious plant growth promoting rhizobacteria Bacillus altitudinis MS16 (GenBank Accession no. MG066459) and its effectiveness in phytopathogen inhibition. B.altitudinis MS16 was evaluated in vitro and it was found to fix 82.80 ± 0.6 µg N ml-1 of culture media, can produce 40.74 ± 0.7 µg ml-1 of IAA, release 7.72 ± 0.9 µg ml-1 of insoluble phosphorus and can secrete hydrolytic enzymes. Apart from potassium and zinc solubilization, it exhibited robust motility and biofilm forming abilities. The strain exhibits remarkable root colonizing efficiency and act strongly against phytopathogens like Colletotrichum gloeosporioides, Sclerotinia sclerotiorum, Corynespora cassiicola, Fusarium verticillioides and Fusarium oxysporum f.sp. Pisi with maximum inhibition against S. sclerotiorum (83.0 %) under in-vitro conditions. Since the strain showed promising activity against S. sclerotiorum (ITCC 4042), ultramicroscopic studies were undertaken to investigate their interaction pattern. Ultramicroscopic observations involving SEM, CLSM and AFM revealed severely altered morphology of MS16 treated S. sclerotiorum mycelia. Leaf detached bioassay accompanied with in planta studies showed potential bio-control efficiency of MS16 with 95.7 ± 2.5 % inhibition obtained in leaf detached assay, 96.5 ± 1.7 % and 98.32 ± 0.56 % inhibition in stem and leaf inoculation against S. sclerotiorum. Furthermore, root colonization resulted in prominent root architectural changes. Seed bacterization showed increase in plant growth attributes over the control under in-vitro and in planta conditions. Therefore, the overall results demonstrate the beneficial role of MS16 for growth promotion in mustard plants while controlling sclerotinia infection in mustard.

Sophorolipid Biosurfactant Can Control Cutaneous Dermatophytosis Caused by Trichophyton mentagrophytes.

Trichophyton mentagrophytes, a zoophilic species, is one of the most frequently isolated dermatophytes in many parts of the world. This study investigated the efficacy of a sophorolipid (SL-YS3) produced by Rhodotorula babjevae YS3 against dermatophytosis caused by T. mentagrophytes. SL-YS3 was characterized by gas chromatography-mass spectrometry (GC-MS) and ultra-performance liquid chromatography, coupled with electrospray mass spectrometry (UPLC-ESI-MS). SL-YS3 comprised of six different fatty acids as the hydrophobic components of constituent congeners and sophorose as the hydrophilic component. Inhibitory effects of purified SL-YS3 against hyphal growth was found to be 85% at a 2 mg ml-1 concentration, and MIC was 1 mg ml-1. Microscopic examination with scanning electron microscopy (SEM), atomic force microscopy, and confocal laser scanning microscopy (CLSM) revealed that SL-YS3 exerts its effect by disrupting cell membrane integrity causing cell death. SL-YS3 was also effective in reducing the biofilms formed by T. mentagrophytes, which was observed spectrophotometrically with crystal-violet staining and further validated with SEM and CLSM studies of treated biofilms. In vivo studies in a mouse model of cutaneous dermatophytosis involving macroscopic observations, percent culture recovery from skin samples, and histopathological studies showed that SL-YS3 could effectively cure the infected mice after 21 days of topical treatment. Terbinafine (TRB) was used as a standard drug in the experiments. We demonstrate, for the first time, the antidermatophytic activity of a sophorolipid biosurfactant. The findings are suggestive that SL-YS3 can be formulated as a novel antifungal compound to treat cutaneous mycoses caused by T. mentagrophytes.

A cost-effective and environmentally sustainable process for phycoremediation of oil field formation water for its safe disposal and reuse.

High volumes of formation water comprising of complex mixture of hydrocarbons is generated during crude oil exploration. Owing to ecotoxicological concerns, the discharge of the formation water without remediation of hydrocarbonaceous pollutants is not permitted. Keeping this into mind, we carried out phycoremediation of hydrocarbons in formation water so that it can be safely discharged or re-used. For this, a native algal species was isolated from formation water followed by its morphological and 18S ribosomal RNA based identification confirming the algal isolate to be Chlorella vulgaris BS1 (NCBI GenBank Accession No. MH732950). The algal isolate exhibited high biomass productivity of 1.76 gm L-1 d-1 (specific growth rate: 0.21 d-1, initial inoculum: 1500 mg L-1) along with remediation of 98.63% petroleum hydrocarbons present in formation water within 14 days of incubation indicating an efficient hydrocarbon remediation process. Concomitantly, the hydrocarbon remediation process resulted in reduction of 75% Chemical Oxygen Demand (COD) load and complete removal of sulfate from formation water making it suitable for safe disposal or reuse as oil well injection water respectively. The present process overcomes the bottlenecks of external growth nutrient addition or dilution associated with conventional biological treatment resulting in a practically applicable and cost-effective technology for remediation of oil field formation water.

Rice based distillers dried grains with solubles as a low cost substrate for the production of a novel rhamnolipid biosurfactant having anti-biofilm activity against Candida tropicalis.

In this study, rhamnolipid (RL) production by Pseudomonas aeruginosa SS14 utilizing rice based Distillers Dried Grains with Solubles (rDDGS) as the sole carbon source was evaluated and the production parameters were optimized using response surface methodology. Highest RL (RL-rDDGS) yield was 14.87 g/L in a culture medium containing 12% (w/v) rDDGS and 11% (v/v) inoculum concentration after 48 h of fermentation at 35 °C. RL-rDDGS was produced as a mixture of mono and di-RL congeners with four novel homologues Rha-C18:2, Rha-C19, Rha-C9, and Rha-Rha-C19. The RL reduced the surface tension of water to 34.8 mN/m at a critical micelle concentration (CMC) value of 100 mg/L, exhibited high stability at a wide range of pH (6-12), heating time (0-120 min), and salinity (2-12% NaCl). Furthermore, RL-rDDGS demonstrated appreciable biofilm disruptive property against Candida tropicalis. This is the first report on the usage of rDDGS for sustainable and low cost production of RL.

Biosurfactant production by a rhizosphere bacteria Bacillus altitudinis MS16 and its promising emulsification and antifungal activity.

Generally, most of the biosurfactant producing microbes are the common inhabitants of hydrocarbon contaminated sites. However, there are a very few literatures that report the existence of biosurfactant producing microbes in rhizosphere soil of plants involved in remediation of pesticide laden agricultural soil and in general welfare of the plants indicating the necessity of further research. A total of twenty three morphologically distinct rhizosphere strains were isolated and were screened for biosurfactant production through preliminary biosurfactant screening assays. Based on the screening results, MS16 was selected as the most efficient and potential biosurfactant producer and was identified as Bacillus altitudinis MS16. Biosurfactant production was indicated by a sharp reduction in the surface tension (ST) of the culture medium from 72.8 to 32.3 ± 0.1 mN/m after 48 h of incubation with a maximum yield of 3.8 g/L. The biosurfactant exhibited excellent emulsifying activity (E24) of 95.4% and emulsion stability of 95.8% against crude oil after an incubation period of 28 days with striking stability over a wide range of temperature (20 - 120 °C, 30 min each), pH (4 - 12) and salinity (2 - 12% NaCl, w/v). The produced biosurfactant was extracted with ethyl acetate and was subjected to compositional analyses revealing it to be a mixed lipopeptide. Furthermore, the biosurfactant exhibited significant antifungal activity showing 42.8% inhibition against Colletotrichum gloeosporioides followed by 41.2% inhibition against Sclerotinia sclerotiorum. To our knowledge, the study reports for the first time biosurfactant producing ability of a novel rhizosphere bacteria Bacillus altitudinis.

Efficacy of a rhamnolipid biosurfactant to inhibit Trichophyton rubrum in vitro and in a mice model of dermatophytosis.

Dermatophytic infections caused by Trichophyton rubrum are the most prevalent superficial mycoses worldwide. The present study was aimed at evaluating the anti-dermatophytic effect of a rhamnolipid biosurfactant (RL) produced by Pseudomonas aeruginosa SS14 (GenBank Accession no. KC866140) against T. rubrum in experimentally induced dermatophytosis in mice models. The purified RL could effectively suppress spore germination and hyphal proliferation of T. rubrum at 500 µg/mL. Ultramicroscopic observations involving SEM and AFM studies revealed severely altered hyphal morphology in the RL-treated mycelia. The membrane disruptive effect of RL was measured by the uptake of propidium iodide (PI) as visualized by CLSM. The absorbance of the RL-treated cell suspension at 260 nm showed concentration-dependant exudation of nucleic acid due to loss of cell membrane integrity. On evaluation of the therapeutic efficacy of RL on experimentally induced cutaneous dermatophytosis in mice models, we observed that the RL, when applied topically at a concentration of 500 µg/mL, was effective in completely curing dermatophytosis at the end of 21-day treatment period. The results were statistically similar to those obtained using the standard drug terbinafine as control. Apart from macroscopic observation, the results were confirmed by culture of skin scrapings and histopathological examination. The results are suggestive of the effectiveness of RL in control of dermatophytosis caused by T. rubrum.

Production, characterization, and antifungal activity of a biosurfactant produced by Rhodotorula babjevae YS3.

BACKGROUND: Sophorolipids are one of the most promising glycolipid biosurfactants and have been successfully employed in bioremediation and various other industrial sectors. They have also been described to exhibit antimicrobial activity against different bacterial species. Nevertheless, previous literature pertaining to the antifungal activity of sophorolipids are limited indicating the need for further research to explore novel strains with wide antimicrobial activity. A novel yeast strain, Rhodotorula babjevae YS3, was recently isolated from an agricultural field in Assam, Northeast India. This study was primarily emphasized at the characterization and subsequent evaluation of antifungal activity of the sophorolipid biosurfactant produced by R. babjevae YS3. RESULTS: The growth kinetics and biosurfactant production by R. babjevae YS3 was evaluated by cultivation in Bushnell-Haas medium containing glucose (10% w/v) as the sole carbon source. A reduction in the surface tension of the culture medium from 70 to 32.6 mN/m was observed after 24 h. The yield of crude biosurfactant was recorded to be 19.0 g/l which might further increase after optimization of the growth parameters. The biosurfactant was characterized to be a heterogeneous sophorolipid (SL) with both lactonic and acidic forms after TLC, FTIR and LC-MS analyses. The SL exhibited excellent oil spreading and emulsifying activity against crude oil at 38.46 mm2 and 100% respectively. The CMC was observed to be 130 mg/l. The stability of the SL was evaluated over a wide range of pH (2-10), salinity (2-10% NaCl) and temperature (at 120 °C for time intervals of 30 up to 120 min). The SL was found to retain surface-active properties under the extreme conditions. Additionally, the SL exhibited promising antifungal activity against a considerably broad group of pathogenic fungi viz. Colletotrichum gloeosporioides, Fusarium verticilliodes, Fusarium oxysporum f. sp. pisi, Corynespora cassiicola, and Trichophyton rubrum. CONCLUSIONS: The study reports, for the first time, the biosurfactant producing ability of R. babjevae, a relatively lesser studied yeast. The persistent surface active properties of the sophorolipid in extreme conditions advocates its applicability in diverse environmental and industrial sectors. Further, antifungal activities against plant and human pathogens opens up possibilities for development of efficient and eco-friendly antifungal agents with agricultural and biomedical applications.

Characterization of Biosurfactant Produced during Degradation of Hydrocarbons Using Crude Oil As Sole Source of Carbon.

Production and spillage of petroleum hydrocarbons which is the most versatile energy resource causes disastrous environmental pollution. Elevated oil degrading performance from microorganisms is demanded for successful microbial remediation of those toxic pollutants. The employment of biosurfactant-producing and hydrocarbon-utilizing microbes enhances the effectiveness of bioremediation as biosurfactant plays a key role by making hydrocarbons bio-available for degradation. The present study aimed the isolation of a potent biosurfactant producing indigenous bacteria which can be employed for crude oil remediation, along with the characterization of the biosurfactant produced during crude oil biodegradation. A potent bacterial strain Pseudomonas aeruginosa PG1 (identified by 16s rDNA sequencing) was isolated from hydrocarbon contaminated soil that could efficiently produce biosurfactant by utilizing crude oil components as the carbon source, thereby leading to the enhanced degradation of the petroleum hydrocarbons. Strain PG1 could degrade 81.8% of total petroleum hydrocarbons (TPH) after 5 weeks of culture when grown in mineral salt media (MSM) supplemented with 2% (v/v) crude oil as the sole carbon source. GCMS analysis of the treated crude oil samples revealed that P. aeruginosa PG1 could potentially degrade various hydrocarbon contents including various PAHs present in the crude oil. Biosurfactant produced by strain PG1 in the course of crude oil degradation, promotes the reduction of surface tension (ST) of the culture medium from 51.8 to 29.6 mN m-1, with the critical micelle concentration (CMC) of 56 mg L-1. FTIR, LC-MS, and SEM-EDS studies revealed that the biosurfactant is a rhamnolipid comprising of both mono and di rhamnolipid congeners. The biosurfactant did not exhibit any cytotoxic effect to mouse L292 fibroblastic cell line, however, strong antibiotic activity against some pathogenic bacteria and fungus was observed.

Uptake of Total Petroleum Hydrocarbon (TPH) and Polycyclic Aromatic Hydrocarbons (PAHs) by Oryza sativa L. Grown in Soil Contaminated with Crude Oil.

The purpose of this study was to determine whether total petroleum hydrocarbon (TPH) and polycyclic aromatic hydrocarbons (PAHs) present in crude oil contaminated sites are transferred to roots, shoots and finally the grains of rice crops (Oryza sativa L.) grown in those sites. Soil was artificially contaminated with crude oil at concentrations of 0, 1000, 5000, 10,000, and 15,000 mg/kg, followed by planting of rice seedlings. After harvest, TPH in plant samples were measured, and it was determined that the uptake of TPH by the plants gradually increased as the concentration of oil in soil increased. Further, from GC-MS analysis, it was observed that PAHs including naphthalene and phenanthrene bioaccumulated in rice plant parts. Vital physico-chemical properties of soil were also altered due to crude oil contamination. Our study revealed that rice plants grown in crude oil polluted sites can uptake TPH including PAHs, thus emphasising the importance of prior investigation of soil condition before cultivation of crops.

Rhamnolipid Biosurfactant against Fusarium verticillioides to Control Stalk and Ear Rot Disease of Maize.

Antifungal activity of rhamnolipids (RLs) has been widely studied against many plant pathogenic fungi, but not against Fusarium verticillioides, a major pathogen of maize (Zea mays L.). F. verticillioides causes stalk and ear rot of maize or asymptomatically colonizes the plant and ears resulting in moderate to heavy crop loss throughout the world. F. verticillioides produces fumonisin mycotoxins, reported carcinogens, which makes the contaminated ears unsuitable for consumption. In this study, the RL produced using glucose as sole carbon source was characterized by FTIR and LCMS analyses and its antifungal activity against F. verticillioides was evaluated in vitro on maize stalks and seeds. Further, the effect of RL on the mycelia of F. verticillioides was investigated by scanning electron microscopy which revealed visible damage to the mycelial structure as compared to control samples. In planta, treatment of maize seeds with a RL concentration of 50 mg l-1 resulted in improved biomass and fruiting compared to those of healthy control plants and complete suppression of characteristic disease symptoms and colonization of maize by F. verticillioides. The study highlights the potential of RLs to be used for an effective biocontrol strategy against colonization of maize plant by F. verticillioides.

Development of an Efficient Bacterial Consortium for the Potential Remediation of Hydrocarbons from Contaminated Sites.

The intrinsic biodegradability of hydrocarbons and the distribution of proficient degrading microorganisms in the environment are very crucial for the implementation of bioremediation practices. Among others, one of the most favorable methods that can enhance the effectiveness of bioremediation of hydrocarbon-contaminated environment is the application of biosurfactant producing microbes. In the present study, the biodegradation capacities of native bacterial consortia toward total petroleum hydrocarbons (TPH) with special emphasis to poly aromatic hydrocarbons were determined. The purpose of the study was to isolate TPH degrading bacterial strains from various petroleum contaminated soil of Assam, India and develop a robust bacterial consortium for bioremediation of crude oil of this native land. From a total of 23 bacterial isolates obtained from three different hydrocarbons contaminated samples five isolates, namely KS2, PG1, PG5, R1, and R2 were selected as efficient crude oil degraders with respect to their growth on crude oil enriched samples. Isolates KS2, PG1, and R2 are biosurfactant producers and PG5, R1 are non-producers. Fourteen different consortia were designed involving both biosurfactant producing and non-producing isolates. Consortium 10, which comprises two Bacillus strains namely, Bacillus pumilus KS2 and B. cereus R2 (identified by 16s rRNA sequencing) has shown the best result in the desired degradation of crude oil. The consortium showed degradation up to 84.15% of TPH after 5 weeks of incubation, as revealed from gravimetric analysis. FTIR (Fourier transform infrared) and GCMS (Gas chromatography-mass spectrometer) analyses were correlated with gravimetric data which reveals that the consortium has removed a wide range of petroleum hydrocarbons in comparison with abiotic control including different aliphatic and aromatic hydrocarbons.

Utilization of Paneer Whey Waste for Cost-Effective Production of Rhamnolipid Biosurfactant.

The present study aimed at isolating rhamnolipid biosurfactant-producing bacteria that could utilize paneer whey, an abundant waste source as sole medium for the production purpose. Pseudomonas aeruginosa strain, SR17, was isolated from hydrocarbon-contaminated soil that could efficiently utilize paneer whey for rhamnolipid production and reduce surface tension of the medium from 52 to 26.5 mN/m. The yield of biosurfactant obtained was 2.7 g/l, upgraded to 4.8 g/l when supplemented with 2 % glucose and mineral salts. Biochemical, FTIR, and LC-MS analysis revealed that extracted biosurfactant is a combination of both mono and di-rhamnolipid congeners. The critical micelle concentration (CMC) was measured to be 110 mg/l. Emulsification activity of the biosurfactant against n-hexadecane, olive oil, kerosene, diesel oil, engine oil, and crude oil were found to be 83, 88, 81, 92, 86, and 100 %, respectively. The rhamnolipid was detected to be non-toxic against mouse fibroblastic cell line L292.

Antifungal properties of rhamnolipid produced by Pseudomonas aeruginosa DS9 against Colletotrichum falcatum.

The rhamnolipid biosurfactant (RL-DS9) extracted from the bacterial strain Pseudomonas aeruginosa DS9 was evaluated for its antifungal activity against Colletotrichum falcatum that causes red rot in sugarcane. The surface tension (ST) reduction, biosurfactant production, and antifungal activity of biosurfactant against C. falcatum were investigated by using the medium with different carbon sources and it was found to be maximum in glucose. Moreover, highest reduction of ST and production of biosurfactant was achieved at 4.5% (w/v) concentration of glucose. The efficacy of RL-DS9 was compared with a commercially available rhamnolipid (RL-R95) using microtitre plate assay. Results showed that at 100 µg ml(-1) concentration RL-DS9 exhibited 86.6% inhibition against C. falcatum spore germination, and in the same concentration RL-R95 showed 83.3% inhibition. From liquid chromatography-mass spectrometry (LC-MS) analysis, it was revealed that only two similar congeners Rha-(C10 ) and Rha-Rha-(C10:1 ) were found to be in common among both the rhamnolipids. In the plant bioassay test, it was noted that red rot incidence was reduced on the sugarcane plants treated with RL-DS9. This is the first report that rhamnolipid biosurfactant produced by Pseudomonas aeruginosa DS9 could be able to control red rot disease of sugarcane caused due to the infection with the fungus Colletotrichum falcatum.