(A) Diameter of the first (R1), second (R2), and third (R3) swarm rings to the consolidation zone and total swarm diameter for P. mirabilis HI4320 on swarm agar buffered with HEPES to pH 5, 6, 7, 8, or 9. (2011) found that both flagella and PMF fimbrae were down regulated, even though they are shown to be important virulence factors in many other studies. Mot plates were incubated at 30°C for ∼16 h, and swimming diameter was measured by using a caliper. mirabilis have the ability to differentiate in elongated, multinucleated, highly flagellated cells, which then allows them to move together over solid surfaces at very high rates. (D) Representative images for two strains that swarmed in response to all cues except arginine. Testing the influence of pH and urea on swarming.The same general strategy described above for the screen was used to test the impact of pH and urea on swarming. The ability to swarm over more viscous or even solid surfaces is restricted to only a few bacterial families: Furmicutes, alpha proteobacteia, and gama proteobacteria. J. Bacteriol 195(2): 368-377. This organism is not usually a pathogen, but does become a problem when it comes into contact with urea in the urinary tract. Antimicrobial Agents and Chemotherapy. 6), indicating that the cues do not promote swarming on LB agar by enhancing the growth rate. Clin. Even though fumarate reductase (FrdA) was not required for swarming under any condition tested (Fig. Geriatr. Metabolism of many amino acids requires TCA cycle intermediates, and our laboratory recently determined that P. mirabilis HI4320 likely uses a complete oxidative TCA cycle and anaerobic respiration for energy during swarming (27). (13) A recent study by Tuson et. They also inhabit other outside environments, and are especially prevalent in hospitals and care facilities. According to the Journal of the Royal Society of Tropical Medicine and Hygiene, symptoms of CAUTIs typically involve the following: high or worsening fever, blood or cloudy urine, pelvic pain/discomfort, frequent urination (when catheters are removed), unexplained malaise, fatigue, or tenderness in the lower abdomen. From there, infection can spread to other parts of the body. Med. However, mutation of gdhA had no effect on swarming in response to any of the cues, indicating that glutamine most likely does not promote swarming by entering the TCA cycle via this route. (2) An individual flagellum can be thought of as a motor, that is coupled to the flow of protons across the membrane to provide energy for rotation. 2012. (A) Swarm colony diameter of P. mirabilis HI4320 on low-salt LB agar supplemented with l-amino acid swarming cues compared to d-amino acids. Swimming motility was assessed by using Mot medium (10 g/liter tryptone, 5 g/liter NaCl) solidified with 0.3% agar. Urease production and robust swarming motility are the two hallmarks of this organism. Abstract. also looked specifically at genes involved in nitrogen assimilation; they found that the genes glnA and gdhA which code for the glutamine synthase and glutamate dehydrogenase, were also up regulated . Proteus mirabilis is well known for using its flagella to swim through liquids or swarm across solid surfaces. Proteus mirabilis is well-known in clinical laboratories and microbiology survey courses as the species that swarms across agar surfaces, overtaking any other species present in the process. (C) Representative images for 10 strains that did not swarm on LB agar but responded to all five cues. Furthermore, while the response to these cues requires specific metabolic pathways, all but malate appear to promote swarming independently of their known roles in biosynthetic pathways. It is widely distributed in soil and water. In the context of CaUTI, P. mirabilis utilizes this process of swarming to migrate along the catheter surface, gaining entry to the bladder and causing painful and sometimes serious complications (3, 14). 2010. Therefore, arginine and malate are optimal under slightly acidic conditions, while the other cues function well across a wide pH range but appear to be best at basic pH. 2013. Urease production and robust swarming motility are the two hallmarks of this organism. Statistical significance was determined by comparing the swarm diameter under each condition to the diameter on plain medium for each strain. One of the most important changes to occur is the substantial increase of flagella along the cell surface. 15. Johnson R.J., Johnston B, Kuskowski A. M. 2012. 73:1082–1089. Arginine decarboxylase and agmatinase therefore appear to be part of the primary pathway for putrescine synthesis under these conditions. 55(5): 905-914. J. This would also explain the finding that malate promotes swarming only on complex medium and not minimal medium, as other metabolic requirements for swarming, such as the presence of amino acids in the medium, are not satisfied in PMSM. Swarming in response to glutamine, histidine, malate, and ornithine generally exhibited a normal dose response, while arginine exhibited a dose response only at concentrations of up to 20 mM, and higher concentrations prevented swarming (see Fig. For studies on swarm agar, exactly 20 ml of medium was dispensed into 100-mm petri dishes instead of 60-mm petri dishes. S1 in the supplemental material) and are referred to as swarming cues. Unlike LB agar, the addition of sodium chloride to this medium does not make it permissive for swarming (data not shown). (2011) isolated P. mirabilis H14320, a strain found in long-term catheterized women. These factors are genetically expressed when necessary during the course of the bacteria’s infection of the urinary tract increasing its virulence. A field guide to bacterial swarming motility. Bien J, Sokolova O, BozkoP. Ten of the isolates exhibited the same general response to all five swarming cues as P. mirabilis HI4320, although the exact swarm pattern and diameter varied between isolates (Fig. April 22, 2013. Characteristic swarming motility 5. To determine the impact of pH on swarming for P. mirabilis HI4320, swarm agar was buffered with 10 mM HEPES and adjusted to pH 5.0, 6.0, 7.0, 8.0, or 9.0 (Fig. Proteus mirabilis Proteus mirabilis is part of the Enterobacteriaceae family. To first determine if aerobic respiration is required for swarming in response to the cues, NaN3 was added to low-salt LB agar at a concentration low enough to permit growth on LB agar but high enough to inhibit growth in broth culture (0.005%, wt/vol) (Fig. Furthermore, the recent development of antimicrobial catheters may also be used to decrease the risk of CA-UTI in hospitalized patients. August 29, 2012. l-Arginine, l-glutamine, dl-histidine, malate, and dl-ornithine promoted swarming on several types of media without enhancing swimming motility or growth rate. If a catheter is necessary for a hospitalized patient, intermittent catheterization may be used in place of long-term or indwelling catheters, thus reducing the risk of bacterial colonization (25). Identification of factors that promote Proteus mirabilis swarming. In Serratia liquefaciens, coordination of swarming requires sensing and integration of several signals, including relative concentrations of amino acids, culture density, surface recognition, and cell-cell interactions (47). This research was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under award numbers R01AI059722 and F32AI102552 and utilized directed metabolomics core services supported by NIH grant DK089503 awarded to the University of Michigan. P. mirabilis swarming is therefore a fascinating and medically-relevant problem that has perplexed scientists since its discovery. Douglas CWI, Bisset KA. All five swarming cues were also capable of promoting swarming when the salt concentration was decreased to 5 mM or lower (data not shown). 1980. Kearns DB, Losick R. 2003. Eighteen strains were capable of swarming on swarm agar and were further tested for responses to the swarming cues. Unsupplemented LB broth and PMSM were pH ∼7.0. Symptoms of catheter-associated urinary tract infections are similar to the symptoms experienced in simple UTIs, however are slightly more severe. The gdhA gene in particular was an unexpected find, and it is believed to help monitor the carbon-nitrogen balance in the bacteria. We thus conclude that initiation of swarming occurs in response to specific cues and that manipulating concentrations of key nutrient cues can signal whether or not a particular environment is permissive for swarming. Statistical significance was determined by comparing mutants to the parental strain under each condition. While many genes and nutrient requirements involved in the swarming process have been identified, few studies have addressed the signals that promote initiation of swarming … Hootan M. T., Bradley F. S., Cardenas D.D. J. Gen. Microbiol. 4C and D). Factors that impact temporal regulation of swarming, swarm speed, or overall swarm pattern have also been identified, such as putrescine and certain fatty acids (23, 24). Amino acid synthesis mutants were also tested for their ability to respond to the swarming cues on low-salt LB agar (Fig. J. Bacteriol 193(13):3286-3292. (2012) used an agar plate-based assay to assess the bacterial counts of four Foley catheters: one Nitrofurantoin lined catheter, and an all silicon control catheter, and one silver alloy-coated catheter and it’s latex-hydrogel control catheter. However, the requirement for glnA and excess glutamine for swarming may represent a new target for prevention of P. mirabilis swarming on catheters. From MicrobeWiki, the student-edited microbiology resource, Swarming Motility: A Locomotive Advantage, Catheter Associated Urinary Tract Infections: CAUTIs, CAUTI Virulence Factors in Proteus Mirabilis: Pearson et. The swarming capability of Proteus mirabilis is important because it is implicated in the pathogenesis of the bacteria and the swarming capability is associated with the bacteria's ability to express virulence factors Proteus mirabilis has a very characteristic bulls-eye appearance on an agar plate due to the regular periodic cycling between the vegetative and swarming st… Sixty out of 180 formulations allowed for the development of at least one swarm ring, but growth was enhanced in only 25 of these 60 formulations. Antimicrob. In some cases, pain medicine may also be prescribed depending on the severity of symptoms. Testing of isogenic mutants revealed that swarming in response to the cues required putrescine biosynthesis and pathways involved in amino acid metabolism. 6. Notably, buffering of the urine prior to the addition of arginine restored growth to a similar level as that in plain urine, and the ureC mutant did not exhibit a growth defect when cultured with arginine. Importantly, none of the five cues dramatically altered normal swarming on permissive medium, although arginine resulted in a slight but statistically significant decrease in the diameter of the second swarm ring (P < 0.001), and glutamine resulted in a slight but statistically significant increase in the diameter of all swarm rings (P = <0.05) (see Fig. The dashed line indicates the swarm colony diameter on unsupplemented LB agar. 56(9): 4960-7972. International journal of Nephrology. Therefore, arginine and histidine biosynthesis is not required for initiation of swarming in response to cues on LB agar, but l-glutamine must be either synthesized or exogenously provided in excess of the ∼0.6 mM present in LB agar to allow for initiation of swarming, regardless of whether or not the medium would normally be permissive for swarming. 9C and D and data not shown). Error bars represent means and standard deviations for three independent experiments with four replicates each. Proceedings of the National Academy of Sciences of the United States of America. One type was Nitrofurazone-coated, while the other was a silver alloy-coated catheter (Figure 4). Interestingly, glutamine allowed for the development of a bull's-eye pattern, malate and ornithine promoted what appeared to be uncoordinated swarms, and swarming in response to arginine or histidine did not always exhibit normal periodicity. Dashed lines indicate average swarm ring diameters for P. mirabilis HI4320. Gibbs K.A, Greenberg E.P, Wenren L.M. A Bioscreen-C automated growth curve analysis system (Growth Curves USA) was utilized to generate growth curves. P. Proteus mirabilis is the third most common etiological factor of urinary tract infection. Thus, P. mirabilis HI4320 initiates swarming on agar plates in response to swarming-specific cues that include one TCA cycle intermediate (malate), three proteinogenic amino acids (arginine, glutamine, and histidine), and two amino acid intermediates of both polyamine synthesis and the urea cycle (arginine and ornithine). However, taken together, the data clearly show that arginine, histidine, and ornithine promote swarming without affecting the growth rate, regardless of the type of medium, and even though glutamine and malate alter the growth rate, this effect is not correlated with a propensity for swarming. Swarming bacteria can produce a range of patterns, and that these patterns may be linked to different environmental conditions. 2). Am. The dashed line indicates the average swarm ring diameter for P. mirabilis HI4320. J. Phys. Pearson M. M. Alexandra, Y. Smith S. Mobley H. 2011. Swarming cues do not enhance swimming motility. 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