Antibacterial activity of silver nanoparticles against Escherichia coli and methicillin-resistant Staphylococcus pseudintermedius is affected by incorporation into carriers for sustained release.

OBJECTIVE: To establish the lowest effective dose of commercially available nanoparticulate silver (AgNP) for antibacterial activity against Escherichia coli (E coli) and methicillin-resistant Staphylococcus pseudintermedius (MRSP), in vitro, and to establish the effect of incorporating AgNP into carriers for sustained release on this antibacterial activity. SAMPLES: Silver nanoparticle dispersion (0.02 mg/mL) composed of citrate-stabilized, spherical, 10 nm diameter nanoparticles in aqueous buffer. PROCEDURES: E coli and MRSP were treated with 0.01 mg/mL AgNP. The highest concentration of bacteria where growth was inhibited by AgNP was selected for treatment with 0.01 mg/mL AgNP incorporated 3 carriers for sustained release: calcium sulfate hemihydrate (CSH) beads, poloxamer 407 gel, and gelatin sponge, respectively. The antibacterial activity of AgNP and AgNP incorporated into carriers for sustained release was compared with a mixed linear effects model. RESULTS: AgNP inhibited bacterial growth at a concentration of 101 for MRSP and 103 for E coli. For MRSP, the treatment group was associated with bacterial growth (P < .001) while the concentration of bacteria and time were not (P = .292 and P = .289, respectively). For E coli, the treatment group and concentration of bacteria were associated with bacterial growth (P < .001 and = .029, respectively) while time was not (P = .095). Poloxamer 407 gel exerted standalone antibacterial activity against both species of bacteria; sponge and CSH beads did not. CLINICAL RELEVANCE: AgNP has antibacterial activity against E coli and MRSP, which can be reduced when incorporated into carriers for sustained release. Poloxamer 407 gel alone and combined with AgNP exerts antibacterial activity against E coli and MRSP.

Evaluation of RapidChek® SELECT™ for the detection of Salmonella spp. in environmental samples from a veterinary hospital.

Nosocomial salmonellosis in hospitalized animals is a recognized hazard, especially in large animal clinics. A standardized culture protocol (SCP) for detecting Salmonella spp. in environmental samples using a 48-h enrichment step results in a 5-day turnaround time for negative results. The RapidChek® SELECT™ Salmonella (RCSS) test system offers detection of organisms in 22-44 h through double enrichment followed by a lateral flow immunoassay. Negative results are reported within 48 h. To determine the most sensitive and rapid method for detecting Salmonella spp. from environmental samples collected at the large animal Purdue Veterinary Hospital (LA-PVH), a preliminary study compared the performance of RCSS and a SCP when testing artificially spiked and naturally contaminated samples. An expanded study analyzed results obtained using the RCSS method to test 872 environmental samples over a 12-month period. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry was chosen as the confirmation method for RCSS-presumptive positive samples. A randomly selected subset of samples received additional confirmation by real-time PCR. Here, we reported the performance data of RCSS in terms of sensitivity, specificity, and positive predictive value using MALDI-TOF results as reference for comparison. We also provide guidelines for reporting results obtained using this system.

Multiplexed histology analyses for the phenotypic and spatial characterization of human innate lymphoid cells.

Innate lymphoid cells (ILCs) emerge in the last few years as important regulators of immune responses and biological processes. Although ILCs are mainly known as tissue-resident cells, their precise localization and interactions with the microenvironment are still unclear. Here we combine a multiplexed immunofluorescence technique and a customized computational, open-source analysis pipeline to unambiguously identify CD127+ ILCs in situ and characterize these cells and their microenvironments. Moreover, we reveal the transcription factor IRF4 as a marker for tonsillar ILC3, and identify conserved stromal landmarks characteristic for ILC localization. We also show that CD127+ ILCs share tissue niches with plasma cells in the tonsil. Our works thus provide a platform for multiparametric histological analysis of ILCs to improve our understanding of ILC biology.

In Situ Maturation and Tissue Adaptation of Type 2 Innate Lymphoid Cell Progenitors.

Innate lymphoid cells (ILCs) are generated early during ontogeny and persist predominantly as tissue-resident cells. Here, we examined how ILCs are maintained and renewed within tissues. We generated a single cell atlas of lung ILC2s and found that Il18r1+ ILCs comprise circulating and tissue-resident ILC progenitors (ILCP) and effector-cells with heterogeneous expression of the transcription factors Tcf7 and Zbtb16, and CD103. Our analyses revealed a continuous differentiation trajectory from Il18r1+ ST2- ILCPs to Il18r- ST2+ ILC2s, which was experimentally validated. Upon helminth infection, recruited and BM-derived cells generated the entire spectrum of ILC2s in parabiotic and shield chimeric mice, consistent with their potential role in the renewal of tissue ILC2s. Our findings identify local ILCPs and reveal ILCP in situ differentiation and tissue adaptation as a mechanism of ILC maintenance and phenotypic diversification. Local niches, rather than progenitor origin, or the developmental window during ontogeny, may dominantly imprint ILC phenotypes in adult tissues.

Innate lymphoid cells in lung infection and immunity.

In recent years, innate lymphoid cells (ILCs) have emerged as key mediators of protection and repair of mucosal surfaces during infection. The lung, a dynamic mucosal tissue that is exposed to a plethora of microbes, is a playground for respiratory infection-causing pathogens which are not only a major cause of fatalities worldwide, but are also associated with comorbidities and decreased quality of life. The lung provides a rich microenvironment to study ILCs in the context of innate protection mechanisms within the airways, unraveling their distinct functions not only in health but also in disease. In this review, we discuss how pulmonary ILCs play a role in protection against viral, parasitic, bacterial, and fungal challenge, along with the mechanisms underlying this ILC-mediated immunity.