The apparition macrophage and Döderlein bacillus is negatively correlated in class I Papanicolaou smear: A morphological examination.

BACKGROUND: Nonspecific vaginitis, also known as Bacterial vaginosis, unlike genital candidiasis and trichomoniasis, is caused by microbiome breakdown. Döderlein's bacillus are gram-positive bacillus that form a microbiome, reproduce in the female vagina after gaining sexual maturity, secrete lactic acid, and prevent the growth of other vaginitis-causing bacteria. Clue cell are squamous epithelial cells with Gardnerella sp. attached to their cell surface. The presence of clue cell is one of the diagnostic criteria for nonspecific vaginitis. Additionally, although macrophages are reported to protect against candidal vaginitis, there are no reports of studies examining the association between macrophages and clue cell. MATERIALS AND METHODS: After re-staining 300 class I specimens by cervical cancer screening with Papanicolaou staining, the appearance of Döderlein's bacillus, macrophages, and clue cell was observed. RESULT: Age group and appearance of Döderlein's bacillus were negatively correlated. The rate of appearance of macrophages was positively correlated with the age group. In people aged 50 years or more, the appearance rate of clue cells was significantly lower in the macrophage appearance group than that in the non-appearance group. CONCLUSION: This study suggested that macrophages, and not Döderlein's bacillus, may play an important role in defense against nonspecific vaginitis.

Antibacterial Effect of Silver Diammine Fluoride on Cariogenic Organisms

AIM: To screen the possible antimicrobial activity of a range of clinically used, silver-based compounds on cariogenic organisms: silver diammine fluoride (SDF), silver fluoride, and silver nitrate. MATERIALS AND METHODS: Preliminary screening disk-diffusion susceptibility tests were conducted on Mueller-Hinton agar plates inoculated with Streptococcus mutans, Lactobacillus acidophilus, and Actinomyces naeslundii, organisms known to be cariogenic. In order to identify which component of the silver compounds was responsible for any antibacterial (AB) effect, and to provide controls, the following were also investigated at high and low concentrations: sodium fluoride, ammonium fluoride, ammonium chloride, sodium fluoride, sodium chloride, and sodium nitrate, as well as deionized water as control. A volume of 10 pL of a test solution was dispensed onto a paper disk resting on the inoculated agar surface, and the plate incubated anaerobically at 37°C for 48 hours. The zones of inhibition were then measured. RESULTS: Silver diammine fluoride, silver fluoride, silver nitrate, and ammonium fluoride had significant AB effect (p < 0.05) on all three test organisms, although ammonium fluoride had no effect at low concentration; the remaining other compounds had no effect. CONCLUSION: Silver ions appear to be the principal AB agent at both high and low concentration; fluoride ions only have an AB effect at high concentration, while ammonium, nitrate, chloride and sodium ions have none. The anticaries effect of topical silver solutions appears restricted to that of the silver ions. CLINICAL SIGNIFICANCE: Silver compounds, such as SDF, silver fluoride, and silver nitrate have AB effect against cariogenic organisms and these may have clinical impact in arresting or preventing dental decay. Sodium fluoride did not have AB effect under the conditions tested.

Photodynamic Inactivation of Cariogenic Pathogens Using Curcumin as Photosensitizer.

OBJECTIVE: This investigation assessed the susceptibility of Streptococcus mutans and Lactobacillus acidophilus to Photodynamic Therapy (PDT) when grown simultaneously in dentine carious lesions. BACKGROUND DATA: PDT is a technique that utilizes light to activate photosensitizers in the presence of oxygen to produce reactive radicals. MATERIALS AND METHODS: A culture medium of 1% glucose, 2% sucrose, 1% young primary culture of L. acidophilus 108 CFU/mL, and S. mutans 108 CFU/mL was utilized to inoculate the bacterial induced caries on human dentine slabs. Different concentrations of the photosensitizer (0.75, 1.5, 3.0, 4.0, and 5.0 g/L) were activated through exposure to the light-emitting diode source with a central wavelength of 450 nm and a fluency of 5.7 J/cm2. Two light intensities (19 and 47.5 mW/cm2) were tested. Four different groups were analyzed: L-D- (control group), L-D+ (drug group), L+D+1 (PDT group 1, light intensity of 19 mW/cm2), and L+D+2 (PDT group 2, light intensity of 47.5 mW/cm2). ANOVA/Tukey tests were utilized to compare groups (α = 5%). RESULTS: Both light intensities required 5.0 g/L of curcumin for significant bacterial reduction (p < 0.05). No significant effect was found for L-D+, thus proving the absence of a potential inherent toxicity. CONCLUSIONS: Curcumin has a toxic effect on microorganisms at appreciable concentrations upon photoactivation. However, it was required to use the maximum concentration of the drug for a successful procedure.

Lactobacillus acidophilus stimulates intestinal P-glycoprotein expression via a c-Fos/c-Jun-dependent mechanism in intestinal epithelial cells.

Our previous studies showed that Lactobacillus acidophilus (LA) culture supernatant (CS) increased P-glycoprotein [Pgp/multidrug resistance 1 (MDR1)] function, expression, and promoter activity in Caco-2 cells. The current studies were designed to elucidate the molecular mechanisms mediating the stimulatory effects of LA CS on Pgp promoter activity. Deletion analysis indicated that the LA CS response element(s) is located in the -172/+428-bp region, and sequence analysis of this region revealed three potential binding sites for c-Fos or c-Jun: proximal activating protein (AP) 1a (-119/-98 bp), distal AP1b (-99/-78 bp), and AP1c (+175/+196 bp). LA CS (24 h) showed an approximately twofold increase in the protein expression of c-Fos and c-Jun in Caco-2 cells. Electrophoretic mobility shift assay showed that LA CS markedly increased the binding of Caco-2 nuclear proteins to AP1a and AP1b, but not AP1c. The DNA-protein complex was completely eliminated by c-Fos antibody, while c-Jun antibody partially eliminated the complex. Chromatin immunoprecipitation analysis also showed that LA CS enhanced the association of c-Fos and c-Jun (by ∼4- and 1.5-fold, respectively) with endogenous Pgp promoter in Caco-2 cells (p-172/+1). Interestingly, overexpression of c-Fos or c-Jun activated Pgp promoter by nearly twofold each. This increase was further enhanced (∼14-fold) when c-Fos and c-Jun were simultaneously overexpressed, suggesting that the presence of one of these transcription factors potentiates the effect of the other. These studies, for the first time, provide evidence for the involvement of c-Fos/c-Jun in stimulation of Pgp gene expression by LA CS in the human intestine.

Formulation and in vivo evaluation of probiotics-encapsulated pellets with hydroxypropyl methylcellulose acetate succinate (HPMCAS).

The aim of this study was to formulate probiotics-encapsulated pellets with hydroxypropyl methylcellulose acetate succinate (HPMCAS) using a dry powder coating technique to improve the storage stability, acid resistance, and intestinal adherence of viable bacteria (Lactobacillus acidophilus and Bifidobacteria animalis ssp. Lactis). Dry coated pellet (DCP) loaded with probiotics was optimized with respect to the quantity of the HPMCAS, an enteric coating polymer (108 mg), and the kinds and amounts of plasticizer (triethyl citrate, 15.7 mg; acetylated monoglyceride, 6.8 mg), by evaluating the survival rate of the bacteria during preparation process and in an acidic medium. Dry coating process allows the whole survivals of living bacteria during preparation process. The DCP formulation exhibited markedly higher acid tolerability and storage stability compared to uncoated viable bacteria. In an in vivo mucosal adherence study in rats, a profound colonization of viable bacteria in the small and large intestine was observed in rats receiving DCP system (p<0.05) compared to rats receiving uncoated probiotics. Moreover, we found that the repeated DCP administration noticeably inhibited intestinal penetration of endotoxin, a potent inflammatory stimulant, from intestinal mucus. The novel DCP system may be an alternative approach for improving bacterial viability in the preparation process and in an acidic medium, and to promote mucosal colonization of probiotic bacteria in the human gut.

The potential risks of probiotics among HIV-infected persons: Bacteraemia due to Lactobacillus acidophilus and review of the literature.

Lactobacillus sp. are commensal organisms that are increasingly reported to cause invasive infections among immunosuppressed persons. However, few data exist regarding the occurrence and risk factors of these infections among HIV-infected persons. Further, the safety of products that contain lactobacilli (e.g. probiotics) in certain populations, including those with HIV/AIDS, is unclear. We report a case of Lactobacillus acidophilus bacteraemia in a patient with AIDS temporally related to excessive consumption of probiotic-enriched yogurt, and provide a comprehensive review of the literature of Lactobacillus sp. infections among HIV-infected persons.

Polyethylene glycol-based hydrogels for controlled release of the antimicrobial subtilosin for prophylaxis of bacterial vaginosis.

Current treatment options for bacterial vaginosis (BV) have been shown to be inadequate at preventing recurrence and do not provide protection against associated infections, such as that with HIV. This study examines the feasibility of incorporating the antimicrobial peptide subtilosin within covalently cross-linked polyethylene glycol (PEG)-based hydrogels for vaginal administration. The PEG-based hydrogels (4% and 6% [wt/vol]) provided a two-phase release of subtilosin, with an initial rapid release rate of 4.0 µg/h (0 to 12 h) followed by a slow, sustained release rate of 0.26 µg/h (12 to 120 h). The subtilosin-containing hydrogels inhibited the growth of the major BV-associated pathogen Gardnerella vaginalis with a reduction of 8 log10 CFU/ml with hydrogels containing ≥15 µg entrapped subtilosin. In addition, the growth of four common species of vaginal lactobacilli was not significantly inhibited in the presence of the subtilosin-containing hydrogels. The above findings demonstrate the potential application of vaginal subtilosin-containing hydrogels for prophylaxis of BV.

Antagonistic activity of Lactobacillus acidophilus LA10 against Salmonella enterica serovar Enteritidis SE86 in mice

Salmonella enterica serovar Enteritidis is one of the main pathogens responsible for foodborne illness in Brazil. Probiotic bacteria can play a role in defense and recovery from enteropathogenic -infections. In this study, the ability of Lactobacillus acidophilus LA10 to colonise and exert anta-gonistic effects in the gastrointestinal tract was tested before and during experimental infection in conventional mice contaminated with S. Enteritidis (SE86). A dose of 0.1 mL containing 10(8) viable cells of SE86 and L. acidophilus LA10 was orally administered by gavage to mice. The experiment was divided into groups. As a negative control, Group 1 was administered only sterile saline solution. As a positive control, Group 2 was administered only SE86. Group 3 was first administered SE86, and after 10 days, treated with L. acidophilus LA10. Group 4 was first administered L. acidophilus LA10,and after 10 days, challenged with SE86.The results demonstrated that a significant number of SE86 cells were able to colonize the gastrointestinal tract of mice, specifically in the colon and ileum. L. acidophilus LA10 demonstrated an antagonistic effect against SE86, with better results observed for Group 3 over Group 4. Thus, L. acidophilus LA10 shows potential antagonistic effects against S. Enteritidis SE86, especially if administered after infection.

Astragalus root and elderberry fruit extracts enhance the IFN-ß stimulatory effects of Lactobacillus acidophilus in murine-derived dendritic cells.

Many foods and food components boost the immune system, but little data are available regarding the mechanisms by which they do. Bacterial strains have disparate effects in stimulating the immune system. In dendritic cells, the gram-negative bacteria Escherichia coli upregulates proinflammatory cytokines, whereas gram-positive Lactobacillus acidophilus induces a robust interferon (IFN)-ß response. The immune-modulating effects of astragalus root and elderberry fruit extracts were examined in bone marrow-derived murine dendritic cells that were stimulated with L. acidophilus or E. coli. IFN-ß and other cytokines were measured by ELISA and RT-PCR. Endocytosis of fluorescence-labeled dextran and L. acidophilus in the presence of elderberry fruit or astragalus root extract was evaluated in dendritic cells. Our results show that both extracts enhanced L. acidophilus-induced IFN-ß production and slightly decreased the proinflammatory response to E. coli. The enhanced IFN-ß production was associated with upregulation of toll-like receptor 3 and to a varying degree, the cytokines IL-12, IL-6, IL-1ß and TNF-α. Both extracts increased endocytosis in immature dendritic cells, and only slightly influenced the viability of the cells. In conclusion, astragalus root and elderberry fruit extracts increase the IFN-ß inducing activity of L. acidophilus in dendritic cells, suggesting that they may exert antiviral and immune-enhancing activity.

Infectious endocarditis caused by Lactobacillus acidophilus in a patient with mistreated dental caries.

We present a rare case of infectious endocarditis caused by Lactobacillus acidophilus in a patient on long-term steroid use for autoimmune hepatitis. In vitro susceptibility-guided antibiotics with benzylpenicillin plus clindamycin and successive mitral annuloplasty resulted in a favorable outcome. Infectious endocarditis was suspected to be a complication of mistreated periodontal infection. Maintenance of oral hygiene is important in immunocompromised patients.

Lipoteichoic acid-deficient Lactobacillus acidophilus regulates downstream signals.

The trillions of microbes residing within the intestine induce critical signals that either regulate or stimulate host immunity via their bacterial products. To better understand the immune regulation elicited by lipoteichoic acid (LTA)-deficient Lactobacillus acidophilus NCFM in steady state and induced inflammation, we deleted phosphoglycerol transferase gene, which synthesizes LTA in L. acidophilus NCFM. In vitro and in vivo experiments were conducted in order to compare the immune regulatory properties of the L. acidophilus strain deficient in LTA (NCK2025) with its wild-type parent (NCK56) in C57BL/6, C57BL/6 recombination-activation gene 1-deficient (Rag1 (-/-)) and C57BL/6 Rag1(-/-)IL-10(-/-) mice. We demonstrate that NCK2025 significantly activates the phosphorylation of Erk1/2 but downregulates the phosphorylation of Akt1, cytosolic group IV PLA2 and p38 in mouse dendritic cells. Similarly, mice treated orally with NCK2025 exhibit decreased phosphorylation of inflammatory signals (Akt1, cytosolic group IV PLA2 or P38) but upregulate Erk1/2-phosphorylation in colonic epithelial cells in comparison with mice treated with NCK56. In addition, regulation of pathogenic CD4+ T cell induced colitis by NCK2025 was observed in Rag1 (-/-) but not Rag1(-/-)IL-10 (-/-) mice suggests a critical role of IL-10 that may be tightly regulated by Erk1/2 signaling. These data highlight the immunosuppressive properties of NCK2025 to deliver regulatory signals in innate cells, which results in the mitigation of T-cell-induced colitis in vivo.

Oligosaccharide-mediated inhibition of the adhesion of pathogenic Escherichia coli strains to human gut epithelial cells in vitro.

The aim of the study was to investigate the ability of pectic oligosaccharides (POS) to inhibit adhesion of three strains of verotoxigenic Escherichia coli, three strains of enteropathogenic E. coli, and one nonclinical strain of Desulfovibrio desulfuricans to human intestinal epithelial cell cultures. Lactobacillus acidophilus and Lactobacillus gasseri were included for comparison. Attachment was determined in the human HT29 cell line by viable count of adherent bacteria. POS in buffer at pH 7.2 were antiadhesive at a dose of 2.5 mg ml(-1), reducing adhesion of enteropathogenic E. coil and verotoxigenic E. coli strains to less than 30% of control values. Concentrations resulting in 50% inhibition ranged from 0.15 to 0.46 mg ml(-1). L. acidophilus was not significantly affected, but adhesion of L. gasseri was reduced to 29% of the control value. POS reduced the adhesion of D. desulfuricans to 0.33% of the control value. POS also had a protective effect against E. coli verocytotoxins VT1 and VT2 at concentrations of 0.01 and 1 microg ml(-1), respectively.

Incipient caries lesions on cementum by mono- and co-culture oral bacteria.

OBJECTIVES: There is increasing prevalence of root caries. We hypothesized different biofilms will cause varying demineralization in cementum. This study investigated the extent of demineralization of cementum by oral biofilm formed from three major cariogenic microorganisms: Streptococcus mutans, Lactobacillus acidophilus, and Actinomyces israelii. Sound cementum tooth blocks were incubated with mono-, bi-, and tri-species combinations of the bacteria under investigation. MATERIALS AND METHODS: The matrix (amide I) and phosphate content of the lesions was analyzed by Fourier-transform infrared spectroscopy (FTIR), and calcium and phosphorus levels were analyzed by scanning electron microscopy and energy-dispersive spectroscopy (SEM-EDX). RESULTS: The log[amide I:HPO(4)(2-) absorbance] values showed that A. israelii mono-culture caused significantly more demineralization than the other bacterial cultures. log[Ca:P] showed that all carious lesions were confined to the cementum. CONCLUSIONS: Oral biofilm arising from bacterial species A. israelii alone was the most cariogenic of those tested and produced the most demineralization in incipient carious lesions in cementum.

Lactobacillus acidophilus as a cause of liver abscess in a NOD2/CARD15-positive patient with Crohn's disease.

INTRODUCTION: The development of a fistula and/or an abscess are major complications in patients with Crohn's disease. An abscess can involve any of the major viscera, including the liver and spleen. CASE REPORT: A 27-year-old male patient with a 6-month history of NOD2/CARD15-positive Crohn's disease with stenosis of the terminal ileum, previously treated with corticosteroids, was admitted with fever, chills, diarrhea, fatigue and cachexia. Diagnostic work-up revealed sepsis, an abscess in the right hepatic lobe, multiple abdominal collections and right-sided pleural effusion. It was felt that his poor general condition prevented surgical intervention and complex conservative treatment was initiated. It consisted of total parenteral nutrition, antibiotics, percutaneous drainage of the liver abscess, and thoracocentesis. Pus and blood cultures showed Lactobacillus acidophilus. There was a gradual improvement in the patient's condition and the liver abscess decreased in diameter from 14 to 2 cm. Aminosalicylates and enteral nutrition were gradually introduced. The patient was discharged after 9 weeks in good general condition. CONCLUSION: Although extremely rarely, L. acidophilus can cause hepatic abscess and sepsis. This can be explained by immunologic incompetence due to corticosteroid treatment of Crohn's disease.

Probiotic lactobacilli and bacteraemia in Stockholm.

The objective of the present study was to determine the incidence of lactobacilli-induced bacteraemia in Stockholm, Sweden, between January 1998 and March 2004, and to identify the possible presence of probiotic strains. Isolated strains were checked for growth on selective lactobacilli-agar and the biochemical profiles were determined. The lactobacilli strains were further analysed with molecular methodologies to ascertain eventual similarities with the probiotic strains Lactobacillus paracasei subsp. paracasei F19 (LMG P 17806), Lactobacillus acidophilus NCFB 1748 and Lactobacillus rhamnosus GG (ATCC 53103). The minimum inhibitory concentrations of selected antimicrobial agents were determined by the agar dilution method. The incidence of bacteraemia cases caused by lactobacilli remained at the same level during the study period and constituted to <1% of the total number of bacteraemia cases each y. L. paracasei subsp. paracasei F19, L. acidophilus NCFB 1748 and L. rhamnosus GG were not identified in any of the samples.

A new assay using surface plasmon resonance (SPR) to determine binding of the Lactobacillus acidophilus group to human colonic mucin.

A new binding assay to investigate the mechanism of adhesion of lactic acid bacteria to the human intestine was established by the surface plasmon resonance technique using a biosensor BIACORE1000. Cells of 26 strains of the Lactobacillus acidophilus group as analytes were eluted onto a sensor chip on which were immobilized biotinylated A-trisaccharide polymer probes having human A-type antigen [(GalNAcalpha1-3(Fucalpha1-2)Gal)-] or human colonic mucin of blood type A (HCM-A) as ligands. In the first screening, high adhesive affinity to the A-trisaccharide BP-probe was observed in L. acidophilus OLL2769, L. crispatus JCM8778, LA205 and LA206. In the second screening, which used HCM-A, only L. acidophilus OLL2769 and L. crispatus JCM8778 were selected as adhesive strains with specific binding ability to human A-antigen. The results indicated that some strains of the L. acidophilus group could recognize and bind the sugar chain of A-antigen structure on HCM.

[Use of molecular methods in food microbiology with the example of probiotic use of lactobacilli]. / Anwendung molekularbiologischer Methoden in der Lebensmittelmikrobiologie am Beispiel probiotisch genutzter Laktobazillen.

The aim of molecular methods in food microbiology is the identification or strain specific differentiation of microorganisms. Identification methods include besides taxonomic purposes also the detection of virulence genes or resistance markers. Strain specific differentiation is used for epidemiological investigations or the quality control of technologically used bacteria. Probiotic strains of the Lactobacillus acidophilus-group were investigated with different molecular methods: for identification proteinfingerprinting and RAPD-PCR (Random Amplified Polymorphic DNA-PCR) were applied, for strain specific differentiation pulsed field gel electrophoresis (PFGE) and again RAPD-PCR were used. The molecular methods applied should be chosen depending on the objective (identification, differentiation) and with respect to the organism to be tested. In case of probiotic lactic acid bacteria like the L. acidophilus-group proteinfingerprinting has proved to be successful for identification and with some limitation also RAPD-PCR. For differentiation PFGE is suitable as well as RAPD-PCR. Those methods differ substantially in their work load and in personal requirements, and show different power of discrimination and reproducibility within and between laboratories. This should be considered while choosing the appropriate method.

Safety and efficacy of probiotic lactobacilli in promoting growth in post-weaning Swiss mice.

Lactobacillus rhamnosus MA27/6B and L. acidophilus MA27/6R are strains used in feed as probiotics. Their safety profiles and growth-stimulating properties were investigated via in vivo studies on young Swiss mice. After repeated administrations of different probiotic preparations in drinking water, safety parameters determined from liver, spleen and total weight remained unchanged. The growth-stimulating properties of viable or dead lactobacilli were studied after supplementation in drinking water. The feed intake (FI), water intake and body weight gain (WG) of the animals were compared to those of control mice. The lactobacilli supplementation of a sub-optimal diet made of barley allow recording of measurable growth performance of mice. It significantly increased WG compared to control groups (P < 0.01), by +28.9% and +31.7% for L. rhamnosus MA27/6B and L. acidophilus MA27/6R, respectively. This WG was correlated with a decrease in the consumption index. The effect of the dose ingested was also investigated: 10(8) lactobacilli CFU/mouse/day produced greater WG than 10(2), 10(4) or 10(6) CFU/mouse/day. No significant differences in growth performance parameters were observed between mice fed with 10(8) cells of viable or nonviable preparations. The mouse assay described could be used as a preliminary criterion when screening candidate probiotics for growth performance properties.

Lactobacillus acidophilus sepsis in a neonate.

Lactobacillus species are non-spore-forming, anaerobic, gram-positive rods that cause disease in immunocompromised adults. Few cases have been described in children. We present the case of a 2-month-old infant who apparently developed Lactobacillus acidophilus sepsis from an infected central venous catheter. Physicians should be aware that although Lactobacillus species rarely cause disease in children, they should be considered a possible pathogen when isolated from the blood of a newborn infant.