Concentrating nutrients and recovering water and energy from source separated urine using osmotic microbial fuel cell.

This work presents the use of osmotic microbial fuel cell (OsMFC), for the first time, to concentrate nutrients and recover water and energy from source separated urine. Four sets of concentration of fresh urine as feed and NaCl as draw were examined: 10% fresh urine vs 0.25 M NaCl; 10% fresh urine vs 2 M NaCl; fresh urine vs 0.25 M NaCl; and fresh urine vs 2 M NaCl. A maximum water flux of 14.27 LMH was attained when 10% of fresh urine and 2 M of NaCl were used as feed and draw solutions, respectively. Additionally, OsMFC concentrates ~99% of TOC, TN, NH4+, and 100% of PO43- and NO3- from urine at the feed side. Polarization studies indicate that the power generation in OsMFC is related to the rate of change of conductivity and the initial conductivity of the anolyte. The maximum (0.12187 W m-3) and minimum power densities (5.3372 × 10-4 W m-3) were obtained for the conditions of fresh urine vs 0.25 M NaCl and 10% fresh urine vs 0.25 M NaCl, respectively. The study shows that OsMFC is an effective pretreatment process to concentrate nutrients from urine by recovering water and energy, simultaneously.

Efficacy of intravesical targeting of novel quorum sensing inhibitor nanoparticles against Pseudomonas aeruginosa biofilm-associated murine pyelonephritis.

Pseudomonas aeruginosa biofilm-associated pyelonephritis is a severe infection that can lead to mortality. There are no strategies that can effectively manage this infection since the pathogenesis is controlled by quorum sensing (QS) regulated virulence and recalcitrant biofilms. QS inhibitors (QSIs) are emerging therapeutics against such infections but are associated with cytotoxicity or low bioactivity. Hence, we developed novel quorum sensing inhibitor loaded nanoparticles (QSINPs) using the biopolymers, chitosan (CS) and dextran sulphate (DS) and were intravesically targeted against biofilm-associated murine pyelonephritis. The in-vivo targeting of QSINPs was confirmed by tracking the fluorescein isothiocyanate (FITC) tagged QSINPs in bladder and kidney of mice. On characterising, the QSINPs showed a size of 685.7 nm with a zeta potential of 37.9 and polydispersity index (PDI) of 0.5. Scanning electron microscopy (SEM) indicated spherical shape and bioactivity assays indicated QSI activity till 8 months. Fourier transform infra-red (FTIR) analysis indicated possibility of isothiocyanate bonding in CS with DS and with QSI. The QSINPs showed excellent in vitro antivirulence activity by reducing the virulence factors and biofilm of P. aeruginosa and in vivo therapeutic efficacy with ciprofloxacin (CIP). Hence, we propose a novel next-generation therapeutic and its appropriate targeting route against biofilm-associated pyelonephritis.

Azithromycin-Ciprofloxacin-Impregnated Urinary Catheters Avert Bacterial Colonization, Biofilm Formation, and Inflammation in a Murine Model of Foreign-Body-Associated Urinary Tract Infections Caused by Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a multifaceted pathogen causing a variety of biofilm-mediated infections, including catheter-associated urinary tract infections (CAUTIs). The high prevalence of CAUTIs in hospitals, their clinical manifestations, such as urethritis, cystitis, pyelonephritis, meningitis, urosepsis, and death, and the associated economic challenges underscore the need for management of these infections. Biomaterial modification of urinary catheters with two drugs seems an interesting approach to combat CAUTIs by inhibiting biofilm. Previously, we demonstrated the in vitro efficacy of urinary catheters impregnated with azithromycin (AZM) and ciprofloxacin (CIP) against P. aeruginosa Here, we report how these coated catheters impact the course of CAUTI induced by P. aeruginosa in a murine model. CAUTI was established in female LACA mice with uncoated or AZM-CIP-coated silicone implants in the bladder, followed by transurethral inoculation of 108 CFU/ml of biofilm cells of P. aeruginosa PAO1. AZM-CIP-coated implants (i) prevented biofilm formation on the implant's surface (P ≤ 0.01), (ii) restricted bacterial colonization in the bladder and kidney (P < 0.0001), (iii) averted bacteriuria (P < 0.0001), and (iv) exhibited no major histopathological changes for 28 days in comparison to uncoated implants, which showed persistent CAUTI. Antibiotic implants also overcame implant-mediated inflammation, as characterized by trivial levels of inflammatory markers such as malondialdehyde (P < 0.001), myeloperoxidase (P < 0.05), reactive oxygen species (P ≤ 0.001), and reactive nitrogen intermediates (P < 0.01) in comparison to those in uncoated implants. Further, AZM-CIP-coated implants showed immunomodulation by manipulating the release of inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α), and IL-10 to the benefit of the host. Overall, the study demonstrates long-term in vivo effectiveness of AZM-CIP-impregnated catheters, which may possibly be a key to success in preventing CAUTIs.

Exploiting the antivirulence efficacy of an ajoene-ciprofloxacin combination against Pseudomonas aeruginosa biofilm associated murine acute pyelonephritis.

The study investigated the in vitro, ex vivo and in vivo efficacy of ajoene and ciprofloxacin (CIP) alone and in combination against Pseudomonas aeruginosa biofilms and biofilm-associated murine acute pyelonephritis. The ajoene-CIP combination exhibited significant greater (p < 0.05) antimotility and biofilm inhibitory effects than those obtained when they were applied individually. The combined action of the agents resulted in a significant increase in serum sensitivity and phagocytic uptake and killing of P. aeruginosa (p < 0.001) compared to the untreated control. Mice groups treated with an ajoene (25 mg kg(-1)) and CIP (30 mg kg(-1) or 15 mg kg(-1)) combination showed a significantly (p < 0.001) reduced bacterial load in the kidney and bladder as compared to that of infected controls and mice treated with solo agents on the fifth day post-infection. The decreased levels of biomarkers and photomicrographs of the kidney tissue of the treated mice showed a reduced severity of damage. Hence, the study highlights the antivirulent and therapeutic efficacy of the ajoene-CIP combination at the minimal dosage of CIP.

Adjuvant effect of cranberry proanthocyanidin active fraction on antivirulent property of ciprofloxacin against Pseudomonas aeruginosa.

Quorum sensing inhibitors (QSIs) act as antivirulent agents since quorum sensing (QS) plays a vital role in regulating pathogenesis of Pseudomonas aeruginosa. However, application of single QSI may not be effective as pathogen is vulnerable to successful mutations. In such conditions, combination of QSIs can be exploited as there can be synergistic or adjuvant action. In the present study, we evaluated the antivirulence efficacy of combination of Vaccinium macrocarpon proanthocyanidin active fraction (PAF) and ciprofloxacin (CIP) at their sub-MICs using standard methods followed by analysis of their mode of action on QS using TLC and molecular docking. There was significant improvement in action of CIP when it was combined with PAF in reducing the QS controlled virulence factors (p < 0.05), motilities and biofilm of P. aeruginosa. TLC profiles of QS signals [(Acyl homoserine lactone (AHL) and Pseudomonas quinolone signal (PQS)] indicated that CIP in combination with PAF, besides showing inhibitory action on production of AHLs, also modulated production and inactivation of PQS. Docking scores also supported the observation. We therefore hypothesize that PAF-CIP combination, having improved anti-virulence property; can be exploited as a potent drug pairing against P. aeruginosa.

Applications of thin-layer chromatography in extraction and characterisation of ajoene from garlic bulbs.

Novel and inexpensive methods of thin-layer chromatography (TLC) were employed for the extraction, characterisation and mechanism of quorum sensing inhibition by ajoene, a component from toluene garlic bulb (Allium sativum L.) extract (TGE). TLC profiling of TGE was carried out using ethyl acetate as solvent. Out of total spots extracted from TLC, four spots exhibited quorum sensing inhibitory (QSI) potential. Among those, spot 5 was identified as Z-ajoene by TLC and confirmed by NMR and MS. HPLC analysis indicated 97.7% purity for purified ajoene. TLC densitometric analysis quantified 221.08 µmol/g of ajoene in TGE and indicated that ajoene is stable at 4°C and at acidic pH. HPTLC profiling showed that ajoene exhibits QSI effect by inhibiting the production of both long-chain acyl homoserine lactones and Pseudomonas quinolone signal (PQS) by P. aeruginosa and also by inactivating PQS molecules.