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      Clearance of Gram-Negative Bacterial Pathogens from the Ocular Surface by Predatory Bacteria

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          Abstract

          It was previously demonstrated that predatory bacteria are able to efficiently eliminate Gram-negative pathogens including antibiotic-resistant and biofilm-associated bacteria. In this proof-of-concept study we evaluated whether two species of predatory bacteria, Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus, were able to alter the survival of Gram-negative pathogens on the ocular surface. Clinical keratitis isolates of Pseudomonas aeruginosa (strain PAC) and Serratia marcescens (strain K904) were applied to the ocular surface of NZW rabbits followed by application of predatory bacteria. At time intervals, surviving pathogenic bacteria were enumerated. In addition, B. bacteriovorus and S. marcescens were applied to porcine organ culture corneas under contact lenses, and the ocular surface was examined by scanning electron microscopy. The ocular surface epithelial layer of porcine corneas exposed to S. marcescens, but not B. bacteriovorus was damaged. Using this model, neither pathogen could survive on the rabbit ocular surface for longer than 24 h. M. aeruginosavorus correlated with a more rapid clearance of P. aeruginosa but not S. marcescens from rabbit eyes. This study supports previous evidence that predatory bacteria are well tolerated by the cornea, but suggest that predatory bacteria do not considerably change the ability of the ocular surface to clear the tested Gram-negative bacterial pathogens from the ocular surface.

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          Most cited references53

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          Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli.

          G Bertani (1951)
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            Risk factors and causative organisms in microbial keratitis.

            To establish the risk factors, causative organisms, levels of antibiotic resistance, patient demographics, clinical presentations, and clinical outcomes of microbial keratitis at a tertiary hospital in Australia. Patients who had a corneal scraping for culture over a 5-year period were identified through the local microbiology database, and a retrospective audit of their medical records was carried out. Clinical information was gathered from medical records, and smear, culture, and antibiotic resistance results were from the microbiology database. An index of disease severity was calculated for each patient from scores for the magnitude of the epithelial defect and anterior-chamber reaction and the location of the lesion. Associations between risk factors for keratitis and variables such as patient demographics, causative organism and antibiotic resistance, disease severity, and outcome were analyzed by using analysis of variance and chi tests with appropriate correction for multiple comparisons. Two hundred fifty-three cases of microbial keratitis in 231 patients were included. Sixty percent of patients were men, and there was a bimodal distribution in the age of presentation. Common risk factors for keratitis were contact lens wear (53; 22%), ocular surface disease (45; 18%), ocular trauma (41; 16%), and prior ocular surgery (28; 11%). Gram stains were positive in 33%, with a sensitivity of 53% and specificity of 89%. Cultures of corneal scrapings were positive in 65% of cases, and Pseudomonas aeruginosa (44; 17%), coagulase-negative staphylococci (22; 9%), Staphylococcus aureus (19; 8%), and fungi (7; 3%) were commonly recovered. P. aeruginosa was more common than other culture results in contact lens-related cases (55% vs. 0%-23%; P < 0.001), and S. aureus was more common than other culture results in ocular surgery-related cases (29% vs. 0%-21%; P < 0.001). Patients with keratitis related to prior ocular surface disease had more severe keratitis at the time of scraping (P = 0.037). Cultures positive for Fusarium, P. aeruginosa, and other Gram-negative organisms had statistically significantly more severe keratitis at the time of scraping, whereas patients with negative cultures had milder keratitis (P = 0.030). Only 2% of all bacterial isolates were resistant to ciprofloxacin, 20% of Gram-positive isolates were resistant to cephalothin, and no Gram-negative isolates were resistant to gentamicin. In this series, the most common risk factor for keratitis was contact lens wear and the most commonly isolated organism was P. aeruginosa.
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              Predatory lifestyle of Bdellovibrio bacteriovorus.

              R Sockett (2008)
              Bdellovibrio species are naturally predatory, small, motile, Deltaproteobacteria that invade the periplasm of other larger gram-negative bacteria, killing and digesting them. Bdellovibrio grows and divides inside the prey cell, in a structure called a bdelloplast, which then lyses, releasing the Bdellovibrio to prey upon more bacteria. This capability makes Bdellovibrio a potential therapeutic agent, but since its discovery in the 1960s it has not been applied in this way. This review considers what is known postgenomically about Bdellovibrio and its predatory lifestyle, drawing also from what was learned by the excellent microbial physiology work of the early Bdellovibrio researchers. Recent work on the diversity and evolution of predatory bdellovibrios, the role of surface structures in predation, and the ongoing questions about how Bdellovibrio switches between axenic and predatory growth and how its predatory activities may be tempered in the wild, as well as suggestions for future research priorities, are discussed.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Antibiotics (Basel)
                Antibiotics (Basel)
                antibiotics
                Antibiotics
                MDPI
                2079-6382
                03 July 2021
                July 2021
                : 10
                : 7
                : 810
                Affiliations
                [1 ]Charles T. Campbell Eye Microbiology Laboratory, Department of Ophthalmology, The Eye and Ear Institute, University of Pittsburgh School of Medicine, 203 Lothrop Street, Room 1020, Pittsburgh, PA 15213, USA; romanowskieg@ 123456upmc.edu
                [2 ]Department of Oral Biology, Rutgers School of Dental Medicine, 110 Bergen St, Newark, NJ 07103, USA; sgupta1109@ 123456gmail.com (S.G.); androulla830@ 123456gmail.com (A.P.); kadourde@ 123456sdm.rutgers.edu (D.E.K.)
                Author notes
                [* ]Correspondence: shanksrm@ 123456upmc.edu ; Tel.: +1-(412)-647-3537
                Author information
                https://orcid.org/0000-0003-4246-3276
                https://orcid.org/0000-0003-3373-0360
                Article
                antibiotics-10-00810
                10.3390/antibiotics10070810
                8300752
                34356731
                2072a6e6-583d-4d61-bf23-e3bd2d2db50d
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 21 May 2021
                : 01 July 2021
                Categories
                Article

                ocular infection,predatory bacteria,bdellovibrio,micavibrio,pseudomonas aeruginosa,serratia marcescens,conjunctivitis,keratitis

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