A disseminated Lactobacillus rhamnosus GG ATCC 53103 infection was suspected in a 6 day-old newborn with intrauterine growth restriction (IUGR) symptoms, treated empirically with antibiotics and given L. rhamnosus GG with the aim of preventing antibiotic-associated gastrointestinal complications. The level of C-reactive protein on day 5 compared with day 2 was increased in spite of negative urine and cerebrospinal fluid cultures. The blood sampled on day 6 was found to be positive for lactobacilli, and the isolate was pre-identified as L. rhamnosus or Lactobacillus casei on day 11. The strain identity was then verified as L. rhamnosus GG through PCR and 16S rRNA sequencing. Genotyping with the rep-PCR and AFLP methods confirmed the 100% genetic similarity for both the strain isolated from patient blood and the probiotic product. The newborn became touch-sensitive, cried a lot, had worsening laboratory test results, and increased inflammation parameters, but no fever was observed. After a further 9 days of antibiotic therapy, blood cultures became negative, and laboratory tests improved on day 25. The patient was discharged from the hospital after 27 days. IUGR with a possible link to L. rhamnosus GG bacteraemia might be a new potential risk group, beside patients with organ failure, immunocompromised status and dysfunctional gut barrier mechanisms, for which safe use of probiotics needs careful attention. Universally accepted or improved guidelines for the safer administration of probiotics in risk groups are urgently needed. This report should not discourage the use of probiotics, but should highlight the need for their careful use in IUGR patients.

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Background: Diphtheria outbreaks occurred in endemic areas and imported and indigenous cases are reported in UE/EEA. Because of the high infectiveness and severity of the disease, early and accurate diagnosis of each suspected case is essential for the treatment and management of the case and close contacts. The aim of the study was to establish simple and rapid testing methods based on Loop-Mediated Isothermal Amplification (LAMP) assay for the detection of Corynebacterium diphtheriae and differentiation between toxigenic and non-toxigenic strains. Methods: Corynebacterium diphtheriae and Corynebacterium ulcerans isolates from the National Institute of Public Health-National Institute of Hygiene collection were used for the development of LAMP assay for the diagnosis of diphtheria and nontoxigenic C. diphtheriae infections. Various colorimetric methods for visualization of results were investigated. Sensitivity and specificity of the assay were examined using a collection of DNA samples from various gram-positive and gram-negative bacteria. Results: The LAMP assay for tox and dtxR genes was developed. The sensitivity and specificity of the assay were calculated as 100%. The detection limit was estimated as 1.42 pg/μl concentration of DNA template when the reaction was conducted for 60 min. However, the detection limit was lowered 10 times for every 10 min of reduction in the time of incubation during the reaction. Positive results were successfully detected colorimetrically using hydroxynaphthol blue, calcein, QuantiFluor, and lateral flow Milenia HybriDetect dipsticks. Conclusion: The assay developed in the study might be applied for point-of-care testing of diphtheria and other C. diphtheriae infections as well as for other infections caused by diphtheria-toxin producing Corynebacterium species. It is highly sensitive, specific, inexpensive, easy to use, and suitable for low-resource settings.

BackgroundCorynebacterium diphtheriae is a re-emerging pathogen in Europe causing invasive infections in vaccinated persons and classical diphtheria in unvaccinated persons. In the presented study we analysed genetic changes in C. diphtheriae isolates collected in Poland from the period before the introduction of the mass anti-diphtheria vaccination to the present time when over 98% of the population is vaccinated.MethodsA total of 62 C. diphtheriae isolates collected in the 1950s–1960s, 1990s and 2000–2016 in Poland were investigated. Examined properties of the isolates included toxigenic status, presence of tox gene, biotype, MLST type (ST) and type of infection.ResultsA total of 12 sequence types (STs) were identified among the analysed C. diphtheriae isolates. The highest variability of STs was observed among isolates from diphtheria and asymptomatic carriers collected in the XX century. Over 95% of isolates collected from invasive and wound infections in 2004–2016 belonged to ST8. Isolates from the XX century represented all four biotypes: mitis, gravis, intermedius and belfanti, but the belfanti biotype appeared only after the epidemic in the 1990s. All except three isolates from the XXI century represented the biotype gravis.ConclusionsDuring a diphtheria epidemic period, non-epidemic clones of C. diphtheriae might also disseminate and persist in a particular area after the epidemic. An increase of the anti-diphtheria antibody level in the population causes not only the elimination of toxigenic strains from the population but may also influence the reduction of diversity of C. diphtheriae isolates. MLST types do not reflect the virulence of isolates. Each ST can be represented by various virulent variants representing various pathogenic capacities, for example toxigenic non-invasive, nontoxigenic invasive and nontoxigenic non-invasive.

Background. Bladder carcinoma is the most common malignancy of the urinary tract. Approximately 75-85% of patients present non-muscle invasive bladder cancer (NMIBC). Standard primary treatment for NIMBC is transurethral resection (TUR) followed by intravesical Bacillus Calmette-Guerin (BCG) immunotherapy. BCG has been accepted as the most effective agent in clinical use against NMIBC. Various BCG substrains are used worldwide for bladder cancer immunotherapy although the impact of used BCG substrain on BCG antitumor capacity is a little investigated. Objectives. The aim of this study was to compare the antitumor capacity and the ability to trigger cytokines production of three BCG substrains by stimulation of the local innate immunity in vitro. Material and Methods. The human bladder cancer cell line T24 was co-cultured with each of the BCG substrains: Moreau, Tice and RIVM alone or with BCG pretreated DCs (dendric cells) and allogenic PBMCs derived from the same donor. The inhibition of T24 cell growth was evaluated by 3 H-thymidine incorporation. Production of Th1 cytokines (IFN-γ, TNF-α, IL-12) and Th2 cytokines (IL-10, IL-4) was measured in cultures of BCG-activated PBMCs by ELISA test.Results. An approximately two-fold inhibition of T24 cell proliferation was observed as a direct cytotoxic effect of tested BCG substrains on T24 cells. However, BCG inhibited the growth of tumor cells mainly by activating the effector cells of innate immunity. About a 10-fold inhibition of T24 cell proliferation was observed when T24 cells were co-cultured with allogenic BCG pretreated DC s and PBMCs derived from the same donor. The PBMCs activated by compared live BCG substrains secreted large amounts of TNF-α and IFN-γ cytokines. Conclusions. Tested BCG substrains had little direct inhibitory effect on T24 cell proliferation. Moreau evolutionarily early BCG substrain showed similar strong, indirect antitumor effects as evolutionarily late BCG substrains Tice and RIVM (Adv Clin Exp Med 2014, 23, 6, 877-884).