Effect of Endosymbiotic Bacteria on Fungal Resistance Toward Heavy Metals.

Most studies on metal removal or tolerance by fungi or bacteria focus on single isolates, without taking into consideration that some fungi in nature may be colonized by endobacteria. To address this knowledge gap, we investigated the tolerance and removal of diverse metals with two fungal species: Linnemannia elongata containing Burkholderia-related endobacteria and Benniella erionia containing Mollicute-related endobacteria. Isogenic lines of both species were generated with antibiotic treatments to remove their respective endobacteria. Experiments involved comparing the isogenic lines and wild type fungi in relation to the minimum inhibitory concentration for the metals, the fungal ability to remove these different metals via atomic adsorption spectroscopy, and the interaction of the metals with specific functional groups of the fungi and fungi-bacteria to determine the role of the bacteria via attenuated total reflection fourier transformed infrared (ATR-FTIR). Finally, we determined the influence of different metal concentrations, associated with moderate and high fungal growth inhibition, on the presence of the endobacteria inside the fungal mycelium via quantitative real-time PCR. Results showed that the presence of the endosymbiont increased B. erionia resistance to Mn2+ and increased the removal of Fe2+ compared to isogenic lines. The absence of the endosymbiont in L. elongata increased the fungal resistance toward Fe2+ and improved the removal of Fe2+. Furthermore, when the bacterial endosymbiont was present in L. elongata, a decrease in the fungal resistance to Ca2+, Fe2+, and Cr6+was noticeable. In the ATR-FTIR analysis, we determined that C-H and C = O were the major functional groups affected by the presence of Cu2+, Mn2+, and Fe2+ for L. elongata and in the presence of Cu2+ and Ca2+ for B. eronia. It is noteworthy that the highest concentration of Pb2+ led to the loss of endobacteria in both L. elongata and B. eronia, while the other metals generally increased the concentration of endosymbionts inside the fungal mycelium. From these results, we concluded that bacterial endosymbionts of fungi can play a fundamental role in fungal resistance to metals. This study provides the first step toward a greater understanding of symbiotic interactions between bacteria and fungi in relation to metal tolerance and remediation.

Copper mining bacteria: Converting toxic copper ions into a stable single-atom copper.

The chemical synthesis of monoatomic metallic copper is unfavorable and requires inert or reductive conditions and the use of toxic reagents. Here, we report the environmental extraction and conversion of CuSO4 ions into single-atom zero-valent copper (Cu0) by a copper-resistant bacterium isolated from a copper mine in Brazil. Furthermore, the biosynthetic mechanism of Cu0 production is proposed via proteomics analysis. This microbial conversion is carried out naturally under aerobic conditions eliminating toxic solvents. One of the most advanced commercially available transmission electron microscopy systems on the market (NeoArm) was used to demonstrate the abundant intracellular synthesis of single-atom zero-valent copper by this bacterium. This finding shows that microbes in acid mine drainages can naturally extract metal ions, such as copper, and transform them into a valuable commodity.

Prevention of infection caused by enteropathogenic E. coli O157:H7 in intestinal cells using enrofloxacin entrapped in polymer based nanocarriers.

Poor bioavailability of antibiotics, toxicity, and development of antibiotic-resistant bacteria jeopardize antibiotic treatments. To circumvent these issues, drug delivery using nanocarriers are highlighted to secure the future of antibiotic treatments. This work investigated application of nanocarriers, to prevent and treat bacterial infection, presenting minimal toxicity to the IPEC-J2 cell line. To accomplish this, polymer-based nanoparticles (NPs) of poly(lactide-co-glycolide) (PLGA) and lignin-graft-PLGA (LNP) loaded with enrofloxacin (ENFLX) were synthesized, yielding spherical particles with average sizes of 111.8 ± 0.6 nm (PLGA) and 117.4 ± 0.9 nm (LNP). The releases of ENFLX from PLGA and LNP were modeled by a theoretical diffusion model considering both the NP and dialysis diffusion barriers for drug release. Biocompatible concentrations of ENFLX, enrofloxacin loaded PLGA(Enflx) and LNP(Enflx) were determined based on examination of bacterial inhibition, toxicity, and ROS generation. Biocompatible concentrations were used for treatment of higher- and lower-level infections in IPEC-J2 cells. Prevention of bacterial infection by LNP(Enflx) was enhanced more than 50% compared to ENFLX at lower-level infection. At higher-level infection, PLGA(Enflx) and LNP(Enflx) demonstrated 25% higher prevention of bacteria growth compared to ENFLX alone. The superior treatment achieved by the nanocarried drug is accredited to particle uptake by endocytosis and slow release of the drug intracellularly, preventing rapid bacterial growth inside the cells.

Microbially-induced mineral scaling in desalination conditions: Mechanisms and effects of commercial antiscalants.

Reverse osmosis (RO) technology is promising in the sustainable production of fresh water. However, expansion of RO use has been hindered by membrane fouling, mainly inorganic fouling known as scaling. Although membrane mineral scaling by chemical means have been investigated extensively, mineral scaling triggered by microbial activity has been largely neglected. In this study, the simultaneous biomineralization of CaCO3 and CaSO4 in the presence of three different microbial communities from fresh water, wastewater, and seawater was investigated. In the presence of either 13 or 79 mM of Ca2+ and SO42- in the media, the fresh water microbial community produced calcite/vaterite and vaterite/gypsum, respectively; the wastewater community produced vaterite and vaterite/gypsum, respectively; and the seawater community produced aragonite in both conditions. The results showed that the concentration of salts and the microbial composition influence the types of precipitates produced. The mechanisms of crystal formation of CaCO3 and gypsum by these communities were also investigated by determining the need for metabolic active cells, the effect of a calcium channel blocker, and the presence of extracellular polymeric substances (EPS). The results showed that metabolically active cells can lead to production of EPS and formation of Ca2+ gradient along the cells through calcium channels, which will trigger formation of biominerals. The prevention of biomineralization by these consortia was also investigated with two common polymeric RO antiscalants, i.e. polyacrylic acid (PAA) and polymaleic acid (PMA). Results showed that these antiscalants do not prevent the formation of the bio-precipitates suggesting that novel approaches to prevent biomineralization in RO systems still needs to be investigated.

Recent advances in graphene-based biosensor technology with applications in life sciences.

Graphene's unique physical structure, as well as its chemical and electrical properties, make it ideal for use in sensor technologies. In the past years, novel sensing platforms have been proposed with pristine and modified graphene with nanoparticles and polymers. Several of these platforms were used to immobilize biomolecules, such as antibodies, DNA, and enzymes to create highly sensitive and selective biosensors. Strategies to attach these biomolecules onto the surface of graphene have been employed based on its chemical composition. These methods include covalent bonding, such as the coupling of the biomolecules via the 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide reactions, and physisorption. In the literature, several detection methods are employed; however, the most common is electrochemical. The main reason for researchers to use this detection approach is because this method is simple, rapid and presents good sensitivity. These biosensors can be particularly useful in life sciences and medicine since in clinical practice, biosensors with high sensitivity and specificity can significantly enhance patient care, early diagnosis of diseases and pathogen detection. In this review, we will present the research conducted with antibodies, DNA molecules and, enzymes to develop biosensors that use graphene and its derivatives as scaffolds to produce effective biosensors able to detect and identify a variety of diseases, pathogens, and biomolecules linked to diseases.