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West Lafayette, IN, United States

Kim H.,Purdue University | Doh I.-J.,Purdue University | Bhunia A.K.,Molecular Food Microbiology Laboratory | King G.B.,Purdue University | Bae E.,Purdue University
Optics Express | Year: 2015

A theoretical model for spectral forward scatter patterns from a bacterial colony based on elastic light scatter is presented. The spectral forward scatter patterns are computed by scalar diffraction theory, and compared with experimental results of three discrete wavelengths (405 nm, 635 nm, and 904 nm). To provide quantitative analysis, spectral dependence of diffraction ring width, gap, maxima, minima, and the first deflection point are monitored. Both model and experiment results show an excellent agreement; a longer wavelength induces a wider ring width, a wider ring gap, a smaller pattern size, and smaller numbers of rings. Further analysis using spatial fast Fourier transform (SFFT) shows a good agreement; the spatial frequencies are increasing towards the inward direction, and the slope is inversely proportional to the incoming wavelength. © 2015 Optical Society of America. Source


Koo O.K.,Molecular Food Microbiology Laboratory | Koo O.K.,University of Arkansas | Aroonnual A.,Molecular Food Microbiology Laboratory | Aroonnual A.,Mahidol University | And 2 more authors.
Journal of Applied Microbiology | Year: 2011

Aims: To investigate the suitability of human Hsp60, a receptor for Listeria adhesion protein (LAP), on paramagnetic beads (PMB) to capture Listeria monocytogenes from food in the presence of other Listeria to facilitate rapid and specific detection of this pathogen. Methods and Results: Commercially available streptavidin-coated PMBs were linked with biotinylated Hsp60 (PMB-Hsp60), and the bacterial capture efficiency from pure culture and meat samples was determined. Capture rate was also compared with the monoclonal antibody (MAb)-C11E9-coated beads (PMB-C11E9) and the commercial Dynabeads anti-Listeria. Captured cells were detected and quantified by plating on selective medium, quantitative real-time PCR (qPCR) and a light-scattering sensor. Overall, all ligand-coated beads had similar capture efficiency (varied from 1·8 to 9·2%) for L. monocytogenes under the conditions employed, and the minimum cell number required to achieve such capture was 10 3CFUml -1. PMB-Hsp60 had significantly greater capture efficiency for pathogenic Listeria (P<0·0001) than the nonpathogenic Listeria. In contrast, PMB-C11E9 and Dynabeads anti-Listeria had similar capture efficiency for both. The efficacy of all PMBs to capture L. monocytogenes in the presence of Listeria innocua from food matrices was compared. Although Dynabeads anti-Listeria had the overall best capture efficiency, PMB-Hsp60 was able to selectively capture L. monocytogenes even in the presence of 10-100-fold more L. innocua cells from enriched meat samples. Conclusions: Data show that the human cell receptor, Hsp60, is suitable for the capture of pathogenic Listeria on PMB in the presence of other Listeria in food. Significance and Impact of the Study: As pathogen interaction with host cells is highly specific, host cell receptors could be used as alternate capture molecules on PMB to aid in specific detection of pathogens. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology. Source


Kim H.,Molecular Food Microbiology Laboratory | Bhunia A.K.,Molecular Food Microbiology Laboratory | Bhunia A.K.,Purdue University
Gut Pathogens | Year: 2013

Background: Listeria adhesion protein (Lap), an alcohol acetaldehyde dehydrogenase (lmo1634) promotes bacterial paracellular translocation through epithelial cell junctions during gastrointestinal phase of infection. Secreted Lap is critical for pathogenesis and is mediated by SecA2 system; however, if strain dependent variation in Lap secretion would affect L. monocytogenes paracellular translocation through epithelial barrier is unknown. Methods. Amounts of Lap secretion were examined in clinical isolates of L. monocytogenes by cell fractionation analysis using Western blot. Quantitative reverse transcriptase PCR (qRT-PCR) was used to verify protein expression profiles. Adhesion and invasion of isolates were analyzed by in vitro Caco-2 cell culture model and paracellular translocation was determined using a trans-well model pre-seeded with Caco-2 cells. Results: Western blot revealed that expression of Lap in whole cell preparation of isolates was very similar; however, cell fractionation analysis indicated variable Lap secretion among isolates. The strains showing high Lap secretion in supernatant exhibited significantly higher adhesion (3.4 - 4.8% vs 1.5 - 2.3%, P < 0.05), invasion and paracellular translocation in Caco-2 cells than the low secreting isolates. In cell wall fraction, Lap level was mostly uniform for both groups, while Lap accumulated in cytosol in low secreting strains indicating that Lap distribution in cellular compartments is a strain-dependent phenomenon, which may be controlled by the protein transport system, SecA2. ΔsecA2 mutants showed significantly reduced paracellular translocation through epithelial barrier (0.48 ± 0.01 vs 0.24 ± 0.02, P < 0.05). qRT-PCR did not show any discernible variation in lap transcript levels in either high or low secreting isolates. Conclusion: This study revealed that secreted Lap is an important determinant in Lap-mediated L. monocytogenes translocation through paracellular route and may serve as an indicator for pathogenic potential of an isolate. © 2013 Kim and Bhunia; licensee BioMed Central Ltd. Source


Kim H.,Purdue University | Bai N.,Purdue University | Bhunia A.K.,Molecular Food Microbiology Laboratory | King G.B.,Purdue University | And 2 more authors.
Journal of Biophotonics | Year: 2013

In order to understand the biophysics behind collective behavior of a bacterial colony, a confocal displacement meter was used to measure the profiles of the bacterial colonies, together with a custom built optical density circuits. The system delivered essential information related to the quantitative growth dynamics (height, diameter, aspect ratio, optical density) of the bacterial colony. For example, the aspect ratio of S. aureus was approximately two times higher than that of E. coli O157:H7, while the OD of S. aureus was approximately 1/3 higher than that of E. coli O157:H7. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source


Amalaradjou M.A.,Molecular Food Microbiology Laboratory | Bhunia A.K.,Molecular Food Microbiology Laboratory
Bioengineered | Year: 2013

Enteric infections account for high morbidity and mortality and are considered to be the fifth leading cause of death at all ages worldwide. Seventy percent of all enteric infections are foodborne. Thus significant efforts have been directed toward the detection, control and prevention of foodborne diseases. Many antimicrobials including antibiotics have been used for their control and prevention. However, probiotics offer a potential alternative intervention strategy owing to their general health beneficial properties and inhibitory effects against foodborne pathogens. Often, antimicrobial probiotic action is non-specific and non-discriminatory or may be ineffective. In such cases, bioengineered probiotics expressing foreign gene products to achieve specific function is highly desirable. In this review we summarize the strategic development of recombinant bioengineered probiotics to control enteric infections, and to examine how scientific advancements in the human microbiome and their immunomodulatory effects help develop such novel and safe bioengineered probiotics. Source

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