Methicillin-Resistant Staphylococcus aureus (MRSA) Detected at Four U.S. Wastewater Treatment Plants

Background: The incidence of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infections is increasing in the United States, and it is possible that municipal wastewater could be a reservoir of this microorganism. To date, no U.S. studies have evaluated the occurrence of MRSA in wastewater.Objective: We examined the occurrence of MRSA and methicillin-susceptible S. aureus (MSSA) at U.S. wastewater treatment plants.Methods: We collected wastewater samples from two Mid-Atlantic and two Midwest wastewater treatment plants between October 2009 and October 2010. Samples were analyzed for MRSA and MSSA using membrane filtration. Isolates were confirmed using biochemical tests and PCR (polymerase chain reaction). Antimicrobial susceptibility testing was performed by Sensititre® microbroth dilution. Staphylococcal cassette chromosome mec (SCCmec) typing, Panton-Valentine leucocidin (PVL) screening, and pulsed field gel electrophoresis (PFGE) were performed to further characterize the strains. Data were analyzed by two-sample proportion tests and analysis of variance.Results: We detected MRSA (n = 240) and MSSA (n = 119) in 22 of 44 (50%) and 24 of 44 (55%) wastewater samples, respectively. The odds of samples being MRSA-positive decreased as treatment progressed: 10 of 12 (83%) influent samples were MRSA-positive, while only one of 12 (8%) effluent samples was MRSA-positive. Ninety-three percent and 29% of unique MRSA and MSSA isolates, respectively, were multidrug resistant. SCCmec types II and IV, the pvl gene, and USA types 100, 300, and 700 (PFGE strain types commonly found in the United States) were identified among the MRSA isolates.Conclusions: Our findings raise potential public health concerns for wastewater treatment plant workers and individuals exposed to reclaimed wastewater. Because of increasing use of reclaimed wastewater, further study is needed to evaluate the risk of exposure to antibiotic-resistant bacteria in treated wastewater.     

Environmentally sustainable route to SiO2@Au–Ag nanocomposites for biomedical and catalytic applications

A facile, sustainable, operationally simple and mild method for the synthesis of SiO₂@Au–Ag nanocomposites (NCs) using Nephrolepis cordifolia tuber extract is described and its catalytic, antibacterial and cytotoxic properties were investigated. The fabricated SiO₂@Au–Ag NCs were well characterized by UV-visible spectroscopy, transmission electron microscopy (TEM), energy-dispersive X-ray (EDX), Fourier transform infrared (FT-IR) spectroscopy, powder X-ray diffraction (XRD), thermogravimetric analysis (TGA) and X-ray photoelectron spectroscopy (XPS) to determine the optical activity, size and morphology, elemental composition, functional groups present, crystallinity, thermal stability and chemical state respectively. The obtained SiO₂@Au–Ag NCs exhibited spherical shape SiO₂ decorated with Au and Ag nanoparticles. The diameter of the SiO₂ nanoparticles ranges from 200–246 with average 3 nm diameter of Au and Ag NPs. Synthetic utility of this protocol has been demonstrated by exploring its effective catalytic activities for the solvent-free amidation of carboxylic acid with a primary amine with excellent yields. Moreover, the synthesized nanocomposite exhibited as noticeable antibacterial effect against Gram negative and Gram positive bacteria and better bio-compatibility against human keratinocytes. Thus, additive free SiO₂@Au–Ag NCs display the potential for catalysis and biomedical applications.

A facile, sustainable, operationally simple and mild method for the synthesis of SiO₂@Au–Ag nanocomposites (NCs) using Nephrolepis cordifolia tuber extract is described and its catalytic, antibacterial and cytotoxic properties were investigated.     

Limited in-depth invasion of Fusobacterium nucleatum into in vitro reconstructed human gingiva

ObjectiveFusobacterium nucleatum is an opportunistic pathogen with a key role in subgingival plaque formation and it is found in increased numbers in periodontally affected sites. This study aimed to investigate the potential of F. nucleatum to penetrate and induce alterations in an in vitro reconstructed human gingival mucosa model.MethodsThree-dimensional (3D) organotypic models of human gingiva were engineered using primary gingival keratinocytes and fibroblasts. The reconstructed tissues were challenged with four different strains of fluorescently labelled F. nucleatum in suspension placed on top of epithelial layers. Confocal laser scanning was used to assess the presence of fusobacteria through the organotypic model. Apoptosis (cleaved caspase-3) and cell proliferation (Ki-67) were evaluated by the use of immunohistochemistry in 3D-tissue models. Quantitative real-time PCR was performed to investigate the mRNA expression for MMP-13 and E-cadherin in both 3D-tissues and monolayers.ResultsF. nucleatum invaded the superficial epithelial layers of gingival 3D-tissue models. Challenged tissues showed accentuated shedding of superficial layers and increased number of cleaved caspase-3 and Ki-67 positive cells than controls, although not statistically significant. Levels of E-cadherin and MMP-13 mRNA were not significantly perturbed in multilayer culture. A variable and disproportionate response of MMP-13 mRNA level resulted in challenged primary keratinocytes in monolayers, compared to multilayer culture.ConclusionThese results indicate that F. nucleatum is able to invade superficially a differentiated, stratified gingival epithelium in vitro and triggers the efficient elimination of bacterial infection through epithelial shredding without causing a permanent damage of the tissue.