Corrigendum: Stochastic Individual-Based Modeling of Bacterial Growth and Division Using Flow Cytometry.

[This corrects the article on p. 2626 in vol. 8, PMID: 29354110.].

Cytological Features Associated with Ureaplasma Urealyticum in Pap Cervical Smear

Purpose: Ureaplasma urealyticum is associated with several obstetric complications and increases the importance of risk management in pregnant women. Furthermore, U. urealyticum has been identified as a cofactor that interacts with human papillomavirus infection in cervical cancer onset. The aim of this study was to assess specific cytological features of U. urealyticum infection in Pap smears to determine whether additional microbiological testing should be performed for pregnant women with a high possibility of U. urealyticum infection. Methods: Liquid-based cytology specimens (LBC) from cervical swabs of a total of 55 women, including 33 pregnant women who were negative for intraepithelial lesion or malignancy (NILM) on Pap testing and with U. urealyticum diagnosed without any other infectious microbes and 22 U. urealyticum-negative controls, were used in this study. We evaluated the localization of U. urealyticum by immunofluorescence, cytological features of secondary changes in squamous cells caused by inflammation, and the specimen background in Pap smears. Results: Based on analysis of Pap smears, a significant relationship was observed between U. urealyticum infection and cannonballs (p < 0.05) as well as predominance of coccoid bacteria (p < 0.05). A large number of U. urealyticum were detected in cannonballs by immunofluorescence. Conclusion: In the present study, cytological features in Pap smears of U. urealyticum infected samples, which have hardly been understood thus far, were assessed. The cytological features included cannonballs and predominance of coccoid bacteria. Our results might help in determining whether additional microbiological testing should be performed for pregnant women with a high possibility of U. urealyticum infection.

Stochastic Individual-Based Modeling of Bacterial Growth and Division Using Flow Cytometry.

A realistic description of the variability in bacterial growth and division is critical to produce reliable predictions of safety risks along the food chain. Individual-based modeling of bacteria provides the theoretical framework to deal with this variability, but it requires information about the individual behavior of bacteria inside populations. In this work, we overcome this problem by estimating the individual behavior of bacteria from population statistics obtained with flow cytometry. For this objective, a stochastic individual-based modeling framework is defined based on standard assumptions during division and exponential growth. The unknown single-cell parameters required for running the individual-based modeling simulations, such as cell size growth rate, are estimated from the flow cytometry data. Instead of using directly the individual-based model, we make use of a modified Fokker-Plank equation. This only equation simulates the population statistics in function of the unknown single-cell parameters. We test the validity of the approach by modeling the growth and division of Pediococcus acidilactici within the exponential phase. Estimations reveal the statistics of cell growth and division using only data from flow cytometry at a given time. From the relationship between the mother and daughter volumes, we also predict that P. acidilactici divide into two successive parallel planes.

Morphological and Bactericidal Effects of Amikacin, Meropenem and Imipenem on Pseudomonas aeruginosa.

BACKGROUND: Pseudomonas aeruginosa might be converted to coccoid bacteria under antibiotic stress. Bacterial conversion would increase resistance to antibiotics due to changes in cell wall crosslink or decreased metabolic activity. Morphology of P. aeruginosa under stress conditions (presence of antibiotics) can be changed to elongated bacilli, U shape and finally coccoid bacteria. Results of several researches showed that coccoid bacteria are one of the most important aspects of drug resistance. It would be the major reason for treatment failure. OBJECTIVES: The aim of this study was to determine in vitro morphological and bactericidal effects of amikacin, meropenem and imipenem on P. aeruginosa isolated from clinical specimens. MATERIALS AND METHODS: Eight P. aeruginosa isolates obtained from clinical samples of burned patients and standard strain ATCC 27853 were used in this study. Isolates were identified by biochemical tests and confirmed by PCR method using ITS specific primer. Minimum inhibitory concentrations (MICs) of three antibiotics were determined by E-test method. Bacteria were exposed to antibiotics at different concentrations. Bacterial morphology in different days was examined by specific microscope and viability of isolates was examined by flow cytometry. RESULTS: All used antibiotics at sub MIC concentration had capability to induce coccoid bacteria. The highest rate of induced coccoid bacteria was 98.2% after 8 days, with contribution of imipenem and meropenem at 2 µg/mL concentration. Amikacin at 4 µg/mL concentration induced lower rate of coccoid bacteria (55.05%). Amikacin had a strong bactericidal effect on coccoid bacteria at 8 µg/mL concentration. Imipenem and meropenem showed very weak bactericidal effect on coccoid bacteria. CONCLUSIONS: Induction of coccoid form of P. aeruginosa may be one of the important reasons for antibiotic treatment failure; therefore, prescribed dose of antibiotics should be carefully managed to prevent increasing antibiotic resistance and coccoid bacteria induction.

Morphological and Bactericidal Effects of Different Antibiotics on Helicobacter pylori.

BACKGROUND: Helicobacter pylori (H. pylori) is a spiral Gram negative bacteria that can transform to the coccoid form in adverse conditions. OBJECTIVES: The aim of this study was to determine the in vitro morphological and bactericidal effects of metronidazole, amoxicillin and clarithromycin on H. pylori. MATERIALS AND METHODS: The standard strain 26695 of H. pylori was cultured on Brucella agar (BA) and the minimum inhibitory concentrations (MICs) of three antibiotics were determined by E-test method. The bacteria were exposed to antibiotics at 1/2 MIC, MIC and 2X MIC concentrations in Brucella broth (BB). Induced coccoid forms were confirmed by Gram staining and light microscopy. The viability of cells as well as the susceptibility of viable coccoids to antibiotics were examined using the flow cytometry method. RESULTS: All of the three antibiotics at sub-MIC induced coccoid forms. The highest rates of coccoids (> 90%) were induced at 0.008 µg/mL concentration (1/2 MIC) of amoxicillin, 72 hours postexposure. Metronidazole and clarithromycin with 1/2 MIC (0.5 and 0.125 µg/mL respectively) induced lower rates of coccoid forms (60% and 40% respectively). Potent bactericidal effects on coccoids were observed with Metronidazole at 2X MIC and clarithromycin at MIC (0.25 µg/mL) (80 - 90%). Amoxicillin with MIC and 2X MIC had no bactericidal effect on coccoid forms. CONCLUSIONS: Despite the good in vitro bactericidal effect of amoxicillin on spiral forms of H. pylori, this antibiotic has little effect on induced coccoids that may develop after the inappropriate in vivo antibacterial treatment. Hence, for successful therapy, it is essential not only to eradicate the spiral forms, but to eliminate the viable coccoids.