scholarly journals Low-Level Tolerance to Fluoroquinolone Antibiotic Ciprofloxacin in QAC-Adapted Subpopulations of Listeria monocytogenes

2021 ◽  
Vol 9 (5) ◽  
pp. 1052
Author(s):  
Divya Kode ◽  
Ramakrishna Nannapaneni ◽  
Mohit Bansal ◽  
Sam Chang ◽  
Wen-Hsing Cheng ◽  
...  

There was a development of low-level tolerance to fluoroquinolone antibiotic ciprofloxacin in Listeria monocytogenes after sublethal adaptation to quaternary ammonium compound (QAC). Using eight L. monocytogenes strains, we determined the changes in short-range MIC, growth rate, and survival for heterologous stress response to ciprofloxacin, after sublethal exposure to daily cycles of fixed or gradually increasing concentration of QAC. Three main findings were observed. (1) MIC increase—QAC-adapted subpopulations exhibited a significant increase in short-range MIC of ciprofloxacin, by 1.5 to 2.9 fold, as compared to non-adapted control for 4/8 strains (p < 0.05). (2) Growth rate increase—QAC-adapted subpopulations exhibited significant 2.1- to 6.8- fold increase in growth rate (OD600 at 10 h) in ciprofloxacin-containing broth, as compared to non-adapted control for 5/8 strains (p < 0.05). (3) Survival increase—QAC-adapted subpopulations of L. monocytogenes yielded significantly higher survival in ciprofloxacin-containing agar by 2.2 to 4.3 log CFU/mL for 4/8 strains, as compared to non-adapted control (p ˂ 0.05). However, for other 4/8 strains of L. monocytogenes, there was no increase in survival of QAC-adapted subpopulations, as compared to non-adapted control in ciprofloxacin. These findings suggest the potential formation of low-level ciprofloxacin-tolerant subpopulations in some L. monocytogenes strains when exposed to residual QAC concentrations (where QAC might be used widely) and such cells if not inactivated might create food safety risk.

2011 ◽  
Vol 79 (12) ◽  
pp. 5001-5009 ◽  
Author(s):  
Patricia A. Spears ◽  
M. Mitsu Suyemoto ◽  
Terri S. Hamrick ◽  
Rebecca L. Wolf ◽  
Edward A. Havell ◽  
...  

ABSTRACTAListeria monocytogenesglcVmutation precludes the binding of certain listerial phages and produces a profound attenuation characterized by the absence of detectable mutants in the livers and spleens of orally inoculated mice.Invitro, we found that the mutant formed plaques on mouse enterocyte monolayers as efficiently as the parent but the plaques formed were smaller. Intracellular growth rate determinations and examination of infected enterocytes by light and fluorescence microscopy established that the mutant was impaired not in intracellular growth rate but in cell-to-cell spreading. Because this property is shared by other immunogenic mutants (e.g.,actAmutants), ourglcVmutant was tested for vaccine efficacy. Oral immunization with the mutant and subsequent oral challenge (22 days postvaccination) with the parent revealed a ca. 10,000-fold increase in protection afforded by the mutant compared to sham-vaccinated controls. TheglcVmutant did not stimulate innate immunity under the dose and route employed for vaccination, and an infectivity index time course experiment revealed pronounced mutant persistence in Peyer's patches. The immunogenicity of theglcVmutant compared to an isogenicactAmutant reference strain was next tested in an experiment with a challenge given 52 days postvaccination. Both mutant strains showed scant vital organ infectivity and high levels of protection similar to those seen using theglcVmutant in the 22-day postvaccination challenge. Our results indicate that oral administration of a profoundly attenuated listerial mutant can safely elicit solid protective immunity.


2011 ◽  
Vol 55 (9) ◽  
pp. 4064-4071 ◽  
Author(s):  
Ellen G. Christensen ◽  
Lone Gram ◽  
Vicky G. Kastbjerg

ABSTRACTThe human food-borne pathogenListeria monocytogenesis capable of persisting in food processing plants despite cleaning and sanitation and is likely exposed to sublethal biocide concentrations. This could potentially affect susceptibility of the bacterium to biocides and other antimicrobial agents. The purpose of the present study was to determine if sublethal biocide concentrations affected antibiotic susceptibility inL. monocytogenes. Exposure ofL. monocytogenesstrains EGD and N53-1 to sublethal concentrations of Incimaxx DES (containing peroxy acids and hydrogen peroxide) and Triquart Super (containing quaternary ammonium compound) in four consecutive cultures did not alter the frequency of antibiotic-tolerant isolates, as determined by plating on 2× the MIC for a range of antibiotics. Exposure of eight strains ofL. monocytogenesto 1 and 4 μg/ml triclosan did not alter triclosan sensitivity. However, all eight strains became resistant to gentamicin (up to 16-fold increase in MIC) after exposure to sublethal triclosan concentrations. Gentamicin-resistant isolates of strains N53-1 and 4446 were also resistant to other aminoglycosides, such as kanamycin, streptomycin, and tobramycin. Gentamicin resistance remained at a high level also after five subcultures without triclosan or gentamicin. Aminoglycoside resistance can be caused by mutations in the target site, the 16S rRNA gene. However, such mutations were not detected in the N53-1-resistant isolates. A combination of gentamicin and ampicillin is commonly used in listeriosis treatment. The triclosan-induced resistance is, hence, of great concern. Further investigations are needed to determine the molecular mechanisms underlying the effect of triclosan.


Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 344
Author(s):  
Md Momtazur Rahman ◽  
Mikhail Vasiliev ◽  
Kamal Alameh

Manipulation of the LED illumination spectrum can enhance plant growth rate and development in grow tents. We report on the identification of the illumination spectrum required to significantly enhance the growth rate of sweet basil (Ocimum basilicum L.) plants in grow tent environments by controlling the LED wavebands illuminating the plants. Since the optimal illumination spectrum depends on the plant type, this work focuses on identifying the illumination spectrum that achieves significant basil biomass improvement compared to improvements reported in prior studies. To be able to optimize the illumination spectrum, several steps must be achieved, namely, understanding plant biology, conducting several trial-and-error experiments, iteratively refining experimental conditions, and undertaking accurate statistical analyses. In this study, basil plants are grown in three grow tents with three LED illumination treatments, namely, only white LED illumination (denoted W*), the combination of red (R) and blue (B) LED illumination (denoted BR*) (relative red (R) and blue (B) intensities are 84% and 16%, respectively) and a combination of red (R), blue (B) and far-red (F) LED illumination (denoted BRF*) (relative red (R), blue (B) and far-red (F) intensities are 79%, 11%, and 10%, respectively). The photosynthetic photon flux density (PPFD) was set at 155 µmol m−2 s−1 for all illumination treatments, and the photoperiod was 20 h per day. Experimental results show that a combination of blue (B), red (R), and far-red (F) LED illumination leads to a one-fold increase in the yield of a sweet basil plant in comparison with only white LED illumination (W*). On the other hand, the use of blue (B) and red (R) LED illumination results in a half-fold increase in plant yield. Understanding the effects of LED illumination spectrum on the growth of plant sweet basil plants through basic horticulture research enables farmers to significantly improve their production yield, thus food security and profitability.


2013 ◽  
Vol 24 (2) ◽  
pp. 115-128 ◽  
Author(s):  
Aaron Z. Welch ◽  
Patrick A. Gibney ◽  
David Botstein ◽  
Douglas E. Koshland

Tolerance to desiccation in cultures of Saccharomyces cerevisiae is inducible; only one in a million cells from an exponential culture survive desiccation compared with one in five cells in stationary phase. Here we exploit the desiccation sensitivity of exponentially dividing cells to understand the stresses imposed by desiccation and their stress response pathways. We found that induction of desiccation tolerance is cell autonomous and that there is an inverse correlation between desiccation tolerance and growth rate in glucose-, ammonia-, or phosphate-limited continuous cultures. A transient heat shock induces a 5000–fold increase in desiccation tolerance, whereas hyper-ionic, -reductive, -oxidative, or -osmotic stress induced much less. Furthermore, we provide evidence that the Sch9p-regulated branch of the TOR and Ras-cAMP pathway inhibits desiccation tolerance by inhibiting the stress response transcription factors Gis1p, Msn2p, and Msn4p and by activating Sfp1p, a ribosome biogenesis transcription factor. Among 41 mutants defective in ribosome biogenesis, a subset defective in 60S showed a dramatic increase in desiccation tolerance independent of growth rate. We suggest that reduction of a specific intermediate in 60S biogenesis, resulting from conditions such as heat shock and nutrient deprivation, increases desiccation tolerance.


2017 ◽  
Vol 80 (3) ◽  
pp. 447-453 ◽  
Author(s):  
Ai Kataoka ◽  
Hua Wang ◽  
Philip H. Elliott ◽  
Richard C. Whiting ◽  
Melinda M. Hayman

ABSTRACT The growth characteristics of Listeria monocytogenes inoculated onto frozen foods (corn, green peas, crabmeat, and shrimp) and thawed by being stored at 4, 8, 12, and 20°C were investigated. The growth parameters, lag-phase duration (LPD) and exponential growth rate (EGR), were determined by using a two-phase linear growth model as a primary model and a square root model for EGR and a quadratic model for LPD as secondary models, based on the growth data. The EGR model predictions were compared with growth rates obtained from the USDA Pathogen Modeling Program, calculated with similar pH, salt percentage, and NaNO2 parameters, at all storage temperatures. The results showed that L. monocytogenes grew well in all food types, with the growth rate increasing with storage temperature. Predicted EGRs for all food types demonstrated the significance of storage temperature and similar growth rates among four food types. The predicted EGRs showed slightly slower rate compared with the values from the U.S. Department of Agriculture Pathogen Modeling Program. LPD could not be accurately predicted, possibly because there were not enough sampling points. These data established by using real food samples demonstrated that L. monocytogenes can initiate growth without a prolonged lag phase even at refrigeration temperature (4°C), and the predictive models derived from this study can be useful for developing proper handling guidelines for thawed frozen foods during production and storage.


1987 ◽  
Vol 50 (5) ◽  
pp. 372-378 ◽  
Author(s):  
ELLIOT T. RYSER ◽  
ELMER H. MARTH

The ability of Listeria monocytogenes to survive the Camembert cheese-making process and grow during ripening of the cheese was examined. Pasteurized whole milk was inoculated to contain about 500 L. monocytogenes [strain Scott A, V7, California, (CA) or Ohio (OH)] CFU/ml and made into Camembert cheese according to standard procedures. All wheels of cheese were ripened at 6°C following 10 d of storage at 15–16°C to allow proper growth of Penicillium camemberti. Duplicate wedge (pie-shaped), surface and interior cheese samples were analyzed for numbers of L. monocytogenes by surface-plating appropriate dilutions made in Tryptose Broth (TB) on McBride Listeria Agar (MLA). Initial TB dilutions were stored at 3°C and surface-plated on MLA after 2, 4, 6 or 8 weeks if the organism was not quantitated in the original sample. Selected Listeria colonies from duplicate samples were confirmed biochemically. Results showed that numbers of Listeria in cheese increased 5- to 10-fold 24 h after its manufacture. Listeria counts for strains Scott A, CA and OH decreased to &lt;10 to 100 CFU/g in all cheese samples taken during the first 18 d of ripening. In contrast, numbers of strain V7 remained unchanged during this period. All L. monocytogenes strains initiated growth in cheese after 18 d of ripening. Maximum Listeria counts of ca. 1 × 106 to 5 × 107 CFU/g were attained after 65 d of ripening. Generally, a 10- to 100-fold increase in numbers of Listeria occurred in wedge or surface as compared to interior cheese samples taken during the latter half of ripening. During this period, Listeria growth paralleled the increase in pH of the cheese during ripening.


1993 ◽  
Vol 70 (3) ◽  
pp. 997-1008 ◽  
Author(s):  
D. C. Lin ◽  
W. Z. Rymer

1. Force changes in areflexive cat soleus muscle in decerebrate cats were recorded in response to two sequential constant velocity (ramp) stretches, separated by a variable time interval during which the length was held constant. Initial (i.e., prestretch) background force was generated by activating the crossed-extension reflex, and stretch reflexes were eliminated by section of ipsilateral dorsal roots. 2. For the initial 400-900 microns of the first stretch, the muscle exhibited high stiffness, classically termed "short-range stiffness." This high stiffness region was followed by an abrupt reduction in stiffness, called muscle "yield," after which force remained at a relatively constant level, achieving a plateau in force. This plateau force level depended largely on stretch velocity, but this dependence was much less than proportional to the increase in stretch velocity, in that a 10-fold increase in velocity produced < 2-fold increase in plateau force. 3. In experiments where the velocities of the two sequential ramp stretches were identical, the force plateau level was the same for each stretch, regardless of the time elapsed before the second stretch (varied from 0 to 500 ms). In contrast, measures of stiffness during the initial portion of the second stretch showed time-dependent magnitude reductions. However, stiffness recovered quickly after the first stretch was completed, returning to control values within 30-40 ms. 4. In one preparation, in which the velocities of the two sequential ramp stretches were different, the force plateau elicited during the second stretch exhibited velocity dependence comparable with that recorded in the earlier single velocity studies. Furthermore, muscle yield was still evident in the case where the force change was due solely to the change in velocity and where short-range stiffness had not yet recovered fully from the initial stretch. On the basis of these findings, we argue that the classical descriptions of short-range stiffness and yield are inadequate and that the change in force that has typically been called the muscle yield reflects a transition between short-range, transient elastic behavior to steady-state, essentially viscous behavior. 5. To examine changes in the muscle's mechanical stiffness during single ramp stretches, a single pulse perturbation was superimposed at various times before, during, and subsequent to the constant velocity stretch. The force increment elicited in response to each pulse decreased relative to the initial isometric value, remained essentially constant until the end of the ramp, and then returned to its prestretch magnitude shortly (30-40 ms) after stretch termination.(ABSTRACT TRUNCATED AT 400 WORDS)


Author(s):  
M. J. A. Mbarga ◽  
I. V. Podoprigora ◽  
E. G. Volina ◽  
A. V. Ermolaev ◽  
L. A. Smolyakova

Introduction: It is already well known that the exposure of certain bacteria, pathogenic or not, to antimicrobials is likely to increase their virulence and induce the development of direct or cross resistance to antimicrobials, but there is almost no information available regarding probiotics. Aim: To assess the changes induced in susceptibility to antibiotics, biofilm formation, growth rate and relative pathogenicity in the probiotic Escherichia coli M17 (EC-M17) after long exposure to antimicrobials namely ampicillin, kanamycin, cefazolin and silver nanoparticles (AgNPs). Methods: After determining the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of the 4 antimicrobials above-mentioned by the microdilution method, EC-M17 was exposed to increasing subinhibitory doses ranging from MIC/8 to MIC for 8 days. The susceptibility to antibiotics of the mutants obtained was assessed by the Kirby Bauer disc diffusion method, biofilm formation by the Congo red agar method and with crystal violet bacterial attachment assay, and relative pathogenicity was assessed using a Galleria melonella waxworm model. Results: Exposure to antimicrobials induces noticeable changes in EC-M17. The highest adaptation to antimicrobials was observed on AgNPs with 8-fold increase in MIC and 16-fold increase in MBC of AgNPs. EC-M17 exposed to ampicillin, kanamycin and silver nanoparticles became resistant to ampicillin, ceftazidime, ceftazidime/clavulanate and tetracycline while exposure to cefazolin induced a significant decrease in sensitivity to tetracycline and ampicillin and resistance to ceftazidime/clavulanate and ceftazidime. The strain exposed to ampicillin was the only one to produce more biofilm than the control strain and except the EC-M17 exposed to cefazolin, all other EC-M17 strains were more pathogenic on G. melonella model than the control. Conclusion: Data in this investigation suggest that repeated exposure of the probiotic EC-M17 to antimicrobials may induce changes in antimicrobials susceptibility, biofilm formation, growth rate, and relative pathogenicity. Therefore, as far as possible, the probiotic E. coli M17 should not be used in combination with antibiotics and further investigations are required to expand similar work on more probiotics in order to avoid resistance build-up which might be transmitted by horizontal transfer.


2020 ◽  
Vol 69 (4) ◽  
pp. 521-529 ◽  
Author(s):  
Matthew E. Wand ◽  
J. Mark Sutton

Introduction. Colistin is a last resort antibiotic for treating infections caused by carbapenem-resistant isolates. Mechanisms of resistance to colistin have been widely described in Klebsiella pneumoniae and Escherichia coli but have yet to be characterized in Citrobacter and Enterobacter species. Aim. To identify the causative mutations leading to generation of colistin resistance in Citrobacter and Enterobacter spp. Methodology. Colistin resistance was generated by culturing in increasing concentrations of colistin or by direct culture in a lethal (above MIC) concentration. Whole-genome sequencing was used to identify mutations. Fitness of resistant strains was determined by changes in growth rate, and virulence in Galleria mellonella. Results. We were able to generate colistin resistance upon exposure to sub-MIC levels of colistin, in several but not all strains of Citrobacter and Enterobacter resulting in a 16-fold increase in colistin MIC values for both species. The same individual strains also developed resistance to colistin after a single exposure at 10× MIC, with a similar increase in MIC. Genetic analysis revealed that this increased resistance was attributed to mutations in PmrB for Citrobacter and PhoP in Enterobacter , although we were not able to identify causative mutations in all strains. Colistin-resistant mutants showed little difference in growth rate, and virulence in G. mellonella, although there were strain-to-strain differences. Conclusions. Stable colistin resistance may be acquired with no loss of fitness in these species. However, only select strains were able to adapt suggesting that acquisition of colistin resistance is dependent upon individual strain characteristics.


2010 ◽  
Vol 67 (7) ◽  
pp. 1057-1067 ◽  
Author(s):  
Paul A. Venturelli ◽  
Nigel P. Lester ◽  
Terry R. Marshall ◽  
Brian J. Shuter

Growing degree-days (GDD, °C·days) are an index of ambient thermal energy that relates directly to an ectotherm’s cumulative metabolism but is rarely used to describe growth and development in fish. We applied GDD to length and maturity data from 416 populations of walleye ( Sander vitreus ) from Ontario and Quebec, Canada (mean annual GDD = 1200 to 2300 °C·days). On average, males matured after they had experienced 6900 °C·days and reached 350 mm total length (L) (n = 77 populations), and females matured after 10 000 °C·days and at 450 mm L (n = 70). Across 143 populations, GDD accounted for up to 96% of the variation in the length of immature walleye but also revealed a twofold difference in growth rate that was indicative of variation in food availability. When applied to data from eight populations in which walleye abundances have changed dramatically over time, GDD revealed a 1.3-fold increase in immature growth rate when abundance was low compared with when it was high. Our results both demonstrate the explanatory power of GDD with respect to fish growth and maturity and inform the development of regional management strategies for walleye.


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