Antimicrobial Applications of Rhamnolipid Biosurfactant Produced from Achromobacter sp. (PS1) Isolate Using Lignocellulosic Hydrolysate

Heterogeneous mixture of partially purified rhamnolipid (RL) produced from Achromobacter sp. (PS1) using lignocellulosic rice straw (RS) sugar hydrolysate medium revealed six different congeners- Rha- C10-C10, Rha-C8-C10/Rha-C10-C8, Rha- C12-C10 / Rha- C10-C12, referring mono-rhamnolipids amounting to total 68.23 % and Rha-Rha-C10-C10, Rha-Rha-C8-C10/Rha-Rha-C10-C8, Rha-Rha-C10-C12/Rha-Rha-C12-C10, referring di-rhamnolipids amounting to 31.73 %, with Mono to Di- RL in the ratio of 2.1:1. This mixture's antimicrobial action containing more mono-rhamnolipids analyzed using broth macro-dilution method exhibited a broad-spectrum antibacterial activity showing ≥ 90 % growth inhibition of both Gram-positive and Gram-negative pathogenic bacteria at MIC ranging from 1.25 mg/mL to 10 mg/mL of total rhamnolipids. This might be due to the more hydrophobic character of mono-rhamnolipids containing a single rhamnosyl group and showing high surface activities. On the other hand, the non-antifungal activity may be attributed to the lower percentage of di-rhamnolipids in the partially purified mixture.

Waste-Based Second-Generation Bioethanol: A Solution for Future Energy Crisis

The demand for more environmentally friendly alternative renewable fuels is growing as fossil fuel resources are depleting significantly. Consequently, bioethanol has attracted interest as a potentially viable fuel. The key steps in second-generation bioethanol production include pretreatment, saccharification, and fermentation. The present study employed simultaneous saccharification and fermentation (SSF) of cellulose through bacterial pathways to generate second-generation bioethanol utilizing corncobs and paper waste as lignocellulosic biomass. Mechanical and chemical pretreatments were applied to both biomasses. Then, two bacterial strains, Bacillus sp. and Norcadiopsis sp., hydrolysed the pretreated biomass and fermented it along with Achromobacter sp., which was isolated and characterized from a previous study. Bioethanol production followed by 72 h of biomass hydrolysis employing Bacillus sp. and Norcadiopsis sp., and then 72 h of fermentation using Achromobacter sp. Using solid phase micro extraction combined with GCMS the ethanol content was quantified. SSF of alkaline pretreated paper waste hydrolysed by Bacillus sp. following the fermentation by Achromobacter sp. showed the maximum ethanol percentage of 0.734±0.154. Alkaline pretreated corncobs hydrolyzed by Norcadiopsis sp. yielded the lowest ethanol percentage of 0.155±0.154. The results of the study revealed that paper waste is the preferred feedstock for generating second-generation bioethanol. To study the possible use of ethanol-diesel blends as an alternative biofuel E2, E5, E7, and E10 blend emulsions were prepared mixing commercially available diesel with ethanol. The evaluated physico-chemical characteristics of the ethanol-diesel emulsions fulfilled the Ceypetco requirements except for the flashpoint revealing that the lower ethanol-diesel blends are a promising alternative to transport fuels. As a result, the current study suggests that second generation bioethanol could be used as a renewable energy source to help alleviate the energy crisis..

Preparation and application of embedded and immobilized Achromobacter sp. agent for effective removal of ammonia nitrogen from water

A novel wastewater-quality-improver, Sodium Alginate Embedded Microbe-treated Zeolite (SAEMZ), was proposed. The strains used are screened from black-odorous water and have high-efficiency NH4+-N degradation performance. The Gram-positive bacteria, belong to Achromobacter sp., was determined through the screening and identification for this strain, whose removal rate of NH4+-N can reach 88.06%, to decrease the NH4+-N's concentration from 61.83 mg/L to 7.80 mg/L, and its optimal growth conditions are pH 7–8, rotation speed 150–210 r/min, temperature 25–35 °C. The SAEMZ's removal effect on NH4+-N were considered in this research from aspects of reusability, storage stability, and the effects of dosage, coexisting ions, and wastewater's concentration. The increase of the SAEMZ's dosage effectively improved NH4+-N's removal rate; Ca2+ in the solution promoted NH4+-N's removal rate, while Mg2+ and Mn2+ inhibited it. Also, NH4+-N's removal rate improved slightly with Fe2+ concentration's increase and then decreased significantly; With the increase of wastewater's dilution factor, NH4+-N's removal rate showed an upward trend and with the increase of the SAEMZ's reuse times, it decreased. Therefore, recycle times should be controlled less than 3 times in practical application; The SAEMZ still maintains its physiological stability, high mechanical strength, and good storage stability after stored at 4 °C for 120 days.

Effect of N-Acetylcysteine in Combination with Antibiotics on the Biofilms of Three Cystic Fibrosis Pathogens of Emerging Importance

Cystic fibrosis (CF) is a genetic disorder causing dysfunctional ion transport resulting in accumulation of viscous mucus that fosters chronic bacterial biofilm-associated infection in the airways. Achromobacter xylosoxidans and Stenotrophomonas maltophilia are increasingly prevalent CF pathogens and while Burkholderia cencocepacia is slowly decreasing; all are complicated by multidrug resistance that is enhanced by biofilm formation. This study investigates potential synergy between the antibiotics ciprofloxacin (0.5–128 µg/mL), colistin (0.5–128 µg/mL) and tobramycin (0.5–128 µg/mL) when combined with the neutral pH form of N-Acetylcysteine (NACneutral) (0.5–16.3 mg/mL) against 11 cystic fibrosis strains of Burkholderia, Stenotrophomonas and Achromobacter sp. in planktonic and biofilm cultures. We screened for potential synergism using checkerboard assays from which fraction inhibitory concentration indices (FICI) were calculated. Synergistic (FICI ≤ 0.5) and additive (0.5 > FICI ≥ 1) combinations were tested on irreversibly attached bacteria and 48 h mature biofilms via time-course and colony forming units (CFU/mL) assays. This study suggests that planktonic FICI analysis does not necessarily translate to reduction in bacterial loads in a biofilm model. Future directions include refining synergy testing and determining further mechanisms of action of NAC to understand how it may interact with antibiotics to better predict synergy.

Phenotype Switching in Metal-Tolerant Bacteria Isolated from a Hyperaccumulator Plant

As an adaptation to unfavorable conditions, microorganisms may represent different phenotypes. Azolla filiculoides L. is a hyperaccumulator of pollutants, but the functions of its microbiome have not been well recognized to date. We aimed to reveal the potential of the microbiome for degradation of organic compounds, as well as its potential to promote plant growth in the presence of heavy metals. We applied the BiologTM Phenotypic Microarrays platform to study the potential of the microbiome for the degradation of 96 carbon compounds and stress factors and assayed the hydrolytic potential and auxin production by the microorganisms in the presence of Pb, Cd, Cr (VI), Ni, Ag, and Au. We found various phenotype changes depending on the stress factor, suggesting a possible dual function of the studied microorganisms, i.e., in bioremediation and as a biofertilizer for plant growth promotion. Delftia sp., Staphylococcus sp. and Microbacterium sp. exhibited high efficacy in metabolizing organic compounds. Delftia sp., Achromobacter sp. and Agrobacterium sp. were efficient in enzymatic responses and were characterized by metal tolerant. Since each strain exhibited individual phenotype changes due to the studied stresses, they may all be beneficial as both biofertilizers and bioremediation agents, especially when combined in one biopreparation.

Microbiome Analysis of the Rhizosphere from Wilt Infected Pomegranate Reveals Complex Adaptations in Fusarium—A Preliminary Study

Wilt disease affecting pomegranate crops results in rapid soil-nutrient depletion, reduced or complete loss in yield, and crop destruction. There are limited studies on the phytopathogen Fusarium oxysporum prevalence and associated genomic information with respect to Fusarium wilt in pomegranate. In this study, soil samples from the rhizosphere of different pomegranate plants showing early stage symptoms of wilt infection to an advanced stage were collected from an orchard situated in Karnataka, India. A whole metagenome sequencing approach was employed to gain insights into the adaptations of the causative pathogen F. oxysporum. Physicochemical results showed a drop in the pH levels, N, Fe, and Mn, and increase in electrical conductivity, B, Zn, Cl, Cu was observed in the early and intermediate stage samples. Comparative abundance analysis of the experimental samples ESI and ISI revealed an abundance of Proteobacteria phyla Achromobacter sp. 2789STDY5608625, Achromobacter sp. K91, and Achromobacter aegrifaciens and Eukaryota namely Aspergillus arachidicola, Aspergillus candidus, and Aspergillus campestris. Functional pathway predictions implied carbohydrate binding to be significant (p < 0.05) among the three experimental samples. Microbiological examination and whole microbiome analysis confirmed the prevalence of F. oxysporum in the soil samples. Variant analysis of F. oxysporum revealed multiple mutations in the 3IPD gene with high impact effects. 3-Isopropylmalate dehydratase and carbohydrate-active enzymes could be good targets for the development of antifungals that could aid in biocontrol of F. oxysporum. The present study demonstrates the capabilities of the whole metagenome sequencing approach for rapid identification of potential key players of wilt disease pathogenesis wherein the symptomatology is complex.

Adaptive Interactions of Achromobacter spp. with Pseudomonas aeruginosa in Cystic Fibrosis Chronic Lung Co-Infection

In the lungs of patients with cystic fibrosis (CF), the main pathogen Pseudomonas aeruginosa is often co-isolated with other microbes, likely engaging in inter-species interactions. In the case of chronic co-infections, this cohabitation can last for a long time and evolve over time, potentially contributing to the clinical outcome. Interactions involving the emerging pathogens Achromobacter spp. have only rarely been studied, reporting inhibition of P. aeruginosa biofilm formation. To evaluate the possible evolution of such interplay, we assessed the ability of Achromobacter spp. isolates to affect the biofilm formation of co-isolated P. aeruginosa strains during long-term chronic co-infections. We observed both competition and cohabitation. An Achromobacter sp. isolate secreted exoproducts interfering with the adhesion ability of a co-isolated P. aeruginosa strain and affected its biofilm formation. Conversely, a clonal Achromobacter sp. strain later isolated from the same patient, as well as two longitudinal strains from another patient, did not show similar competitive behavior against its P. aeruginosa co-isolates. Genetic variants supporting the higher virulence of the competitive Achromobacter sp. isolate were found in its genome. Our results confirm that both inter-species competition and cohabitation are represented during chronic co-infections in CF airways, and evolution of these interplays can happen even at the late stages of chronic infection.