Oral Lactobacillus Research Articles

Effects of antibiotics at sub-minimal inhibitory concentrations on the morphology of Streptococcus mutans and Lactobacillus acidophilus

Sub-minimal inhibitory concentrations (sub-MICs) of antibiotics may have significant impact on the morphology and biochemical characteristics of target bacteria. This study aimed to examine the morphological changes to sub-MICs of antibiotics observed in bacterial cells from Gram-positive oral species, Streptococcus mutans and Lactobacillus acidophilus. The MICs for both amoxicillin and doxycycline were determined by broth dilution method. Individual cells of S . mutans increased in length after incubation with sub-MIC of amoxicillin, whereas its chain length increased in response to a sub-MIC of doxycycline. The lengths of individual bacterial cells of L. acidophilus decreased after incubation with sub-MICs of either amoxicillin or doxycycline. These results of suggest that sub- MICs of amoxicillin and doxycycline induce morphological changes in both S. mutans and L. acidophilus. Further studies are required to determine the significance of these findings for host-pathogen interactions and development of dental diseases.

Safety assessment of the candidate oral probiotic Lactobacillus crispatus YIT 12319: Analysis of antibiotic resistance and virulence-associated genes

Lactobacillus crispatus YIT 12319 (LcY) was isolated from the oral cavity of a healthy subject as a new candidate probiotic with potential benefits for oral health. As a safety assessment of LcY, we performed an antibiotic susceptibility test and virulence-associated gene analysis using a draft genome sequence. Susceptibility to 15 antibiotics was analyzed according to the standard method of the International Dairy Federation/International Organization for Standardization, as recommended by the European Food Safety Authority. The results showed that the minimum inhibitory concentrations of LcY were not higher than those of other L. crispatus strains, which have not acquired resistance to any antibiotics, suggesting that LcY had no externally acquired transmissible antibiotic resistance genes. Analysis of virulence-associated genes using the draft genome of LcY found that there were fewer potential virulence-associated genes in LcY than in other probiotics. These findings suggest that LcY could be a candidate probiotic based on its safety profile.

Study of oral lactobacillus towards developing a comprehensive structured for integrated exponential regression model

Introduction: Probiotics are well-defined as live microorganisms that usefully affect the host and probiotic bacteria have been used intensely. For years to target gastrointestinal disease by rebalancing the compound microflora. Besides the gastrointestinal tract also the oral cavity is highly colonized by bacteria and many different bacterial species are part of the microbiota in the mouth, as it offers ideal conditions for bacteria with a stable temperature, moist surface with a relatively stable pH and regular supply of nutrients. Probiotic bacteria like Lactobacillus are a promising treatment strategy for oral disease with a microbiological etiology. To gain better results, many researchers that study and emphasize specific methods been tried to build a new or improved methodology.
 Objectives: The aimed of this study is to improve the performance of exponential growth by adding bootstrap and fuzzy techniques (Integrated exponential regression method). The aim of the research work is to develop a comprehensive framework for an integrated exponential regression model.
 Material and Methods: The data were taken from the present data available from the recently done by a researcher for nurturing selected microorganisms. The gathered data will be used for the exponential modeling and the efficiency of the model will be compared accordingly due to the predicted interval from the exponential regression method and an integrated exponential regression method. This paper also provides the algorithm for the prediction of cell growth and inferences.
 Results: The result shows that the average width for the exponential regression model was 19.2228 while an integrated exponential regression method was 0.0075. The average width of integrated exponential regression was smaller than the exponential regression. This clearly shows that the integrated exponential regression method is more efficient than exponential regression technique.
 Conclusion: This proposed method can be applied to small sample size data, especially when limited data is obtained.
 Bangladesh Journal of Medical Science Vol.19(3) 2020 p.552-557

Adhesion, anti-adhesion and aggregation properties relating to surface charges of selected Lactobacillus strains: study in Caco-2 and H357 cells.

This study aimed to assess adhesion and anti-adhesion, aggregation, and surface properties of four selected oral Lactobacillus strains, L. fermentum SD7, L. paracasei SD1, L. rhamnosus SD4, and L. rhamnosus SD11, together with Lactobacillus rhamnosus GG. Human cells, enterocytes Caco-2 and oral keratinocyte H357 were used, and various enteric and oral pathogens were included. Results showed that all Lactobacillus tested gave high adhesion and internalization in both Caco-2 and H357 cells similar to L. rhamnosus GG, and it suggests that such properties are strain dependent and specific to host cells. Anti-adhesion was different; it depended on the internalization ability of individual Lactobacillus and pathogenic strains to Caco-2 and H357. Coaggregation ability depended on autoaggregation of both the Lactobacillus and pathogenic strains. A positive correlation between surface charges and aggregation, and internalization and anti-adhesion of all Lactobacillus was found.In conclusion, results suggests that the selected Lactobacillus might be potential probiotics for usage in both the oral cavity and intestinal tract due to their abilities of aggregation, adherence and anti-internalization to both Caco-2 and H357 cells.

Post-antibiotic gut dysbiosis-induced trabecular bone loss is dependent on lymphocytes.

Recent studies in mouse models have shown that gut microbiota significantly influences bone health. We demonstrated that 2-week oral treatment with broad spectrum antibiotics followed by 4weeks of recovery of the gut microbiota results in dysbiosis (microbiota imbalance)-induced bone loss in mice. Because gut microbiota is critical for the development of the immune system and since both microbiota and the immune system can regulate bone health, in this study, we tested the role of the immune system in mediating post-antibiotic dysbiosis-induced bone loss. For this, we treated wild-type (WT) and lymphocyte deficient Rag2 knockout (KO) mice with ampicillin/neomycin cocktail in water for 2weeks followed by 4weeks of water without antibiotics. This led to a significant bone loss (31% decrease from control) in WT mice. Interestingly, no bone loss was observed in the KO mice suggesting that lymphocytes are required for dysbiosis-induced bone loss. Bray-Curtis diversity metrics showed similar microbiota changes in both the WT and KO post-antibiotic treated groups. However, several operational taxonomic units (OTUs) classified as Lactobacillales were significantly higher in the repopulated KO when compared to the WT mice, suggesting that these bacteria might play a protective role in preventing bone loss in the KO mice after antibiotic treatment. The effect of dysbiosis on bone was therefore examined in the WT mice in the presence or absence of oral Lactobacillus reuteri treatment for 4weeks (post-ABX treatment). As hypothesized, mice treated with L. reuteri did not display bone loss, suggesting a bone protective role for this group of bacteria. Taken together, our studies elucidate an important role for lymphocytes in regulating post-antibiotic dysbiosis-induced bone loss.

Effect of Oral Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on the Vaginal Microbiota, Cytokines and Chemokines in Pregnant Women.

Spontaneous preterm birth is associated with vaginal microbial dysbiosis. As certain strains of lactobacilli help restore homeostasis in non-pregnant women, the goal was to determine the effect of Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 administered orally, twice daily for 12 weeks on the vaginal microbiota, cytokines and chemokines of low-risk pregnant women. A double-blind, placebo-controlled, randomized trial comparing probiotic lactobacilli to placebo daily was performed in 86 asymptomatic pregnant women who had an Intermediate or Bacterial Vaginosis Nugent score at 13 weeks. After drop outs, 32 women receiving probiotics and 34 receiving placebo completed the study. The Nugent score returned to normal in 30% of the women in both groups at 28 weeks and was maintained until 35 weeks. The majority of subjects had normal pregnancy outcomes. Ninety-three bacterial species were detected at 13 weeks, with Lactobacillus iners, Lactobacillus crispatus, Gardnerella vaginalis and Atopobium vaginae being the most abundant across pregnancy. There was no difference in the Shannon diversity index between the probiotic and placebo groups at 13, 28 or 35 weeks. Almost all subjects consumed fermented foods and many of the organisms in the vagina are also known to be present in fermented foods. Interleukin-4 in the placebo group and Interleukin-10 in both probiotic and placebo groups increased slightly at 28 weeks but were not different at 35 weeks when compared to 13 weeks. In conclusion, this study showed no adverse issues resulting from 12 week use of probiotic Lactobacillus strains GR-1 and RC-14 during pregnancy in women at low risk for premature birth. The vaginal microbiota demonstrated flux irrespective of this oral probiotic administration.

Construction and immune efficacy of recombinant Lactobacillus casei strains expressing Malt from Aeromonas veronii

Aeromonas veronii is an important zoonotic pathogen that causes significant economic losses in the aquaculture industry. The use of probiotics in aquaculture is a practical alternative to antibiotics to promote animal health and aid in disease prevention. In the present study, we aimed to construct a recombinant Lactobacillus casei(surface-displayed or secretory) strain containing Malt from A. veronii TH0426 and assess its potential as an oral vaccine. A 1314-bp Malt gene fragment was successfully amplified and cloned into a prokaryotic protein expression system. Protein expression in resulting recombinant strains Lc-MCS-Malt (surface-displayed) and Lc-pPG-Malt (secretory) was then verified by Western blotting and indirect immunofluorescence. A single band was observed on the Western blots, with the molecular weight of the corresponding protein shown to be 48 kDa. Furthermore, a fluorescent signal for Lc-MCS-Malt was observed by fluorescence microscopy. At 0, 7, 16, 25, and 34 days post-immunization, tissue and blood samples were collected from common carp orally administered with the recombinant L. casei strains for immune-related index analyses. Treatment of common carp with the recombinant vaccine candidate stimulated high serum or skin mucus specific antibody titers and induced a higher lysozyme, ACP, SOD activity, while fish fed with Lc-pPG or PBS had no detectable immobilizing immune responses. Expression of IL-10, IL-1β, TNF-α, and IFN-γ genes in the group immunized with recombinant L. casei were significantly (P < 0.05) up regulated as compared with control groups, indicating that inflammatory response and cell immune response were triggered. Results also showed that recombinant L. casei could stimulate the mucosa through colonization of the intestine, resulting in increased transcription of IL-10, IL-1β, TNF-α, and IFN-γ. Immunity and colonization assays also showed that after 34 days of fasting, recombinant L. casei were still present in the intestines of the immunized fish. Common carp that received Lc-MCS-Malt(53.3%) and Lc-pPG-Malt (46.7%) exhibited higher survival rates than the controls after challenge with the pathogen A. veronii. Our findings suggested that recombinant L. casei can adequately protect fish and improve immunity, providing a theoretical basis for the future development of an oral Lactobacillus vaccine for use in aquaculture.

Impact of oral probiotic Lactobacillus acidophilus vaccine strains on the immune response and gut microbiome of mice.

The potential role of probiotic bacteria as adjuvants in vaccine trials led to their use as nonparenteral live mucosal vaccine vectors. Yet, interactions between these vectors, the host and the microbiome are poorly understood. This study evaluates impact of three probiotic, Lactobacillus acidophilus, vector strains, and their interactions with the host’s immune response, on the gut microbiome. One strain expressed the membrane proximal external region from HIV-1 (MPER). The other two expressed MPER and either secreted interleukin-1ß (IL-1ß) or expressed the surface flagellin subunit C (FliC) as adjuvants. We also used MPER with rice bran as prebiotic supplement. We observed a strain dependent, differential effect suggesting that MPER and IL-1β induced a shift of the microbiome while FliC had minimal impact. Joint probiotic and prebiotic use resulted in a compound effect, highlighting a potential synbiotic approach to impact efficacy of vaccination. Careful consideration of constitutive adjuvants and use of prebiotics is needed depending on whether or not to target microbiome modulation to improve vaccine efficacy. No clear associations were observed between total or MPER-specific IgA and the microbiome suggesting a role for other immune mechanisms or a need to focus on IgA-bound, resident microbiota, most affected by an immune response.

Recombinant Lactobacillus casei Expressing Capsid Protein VP60 can Serve as Vaccine Against Rabbit Hemorrhagic Disease Virus in Rabbits.

Rabbit hemorrhagic disease virus (RHDV) is the causative agent of rabbit hemorrhagic disease (RHD). RHD, characterized by hemorrhaging, liver necrosis, and high morbidity and mortality in rabbits and hares, causes severe economic losses in the rabbit industry worldwide. Due to the lack of an efficient in-vitro propagation system for RHDV, the current vaccine is produced via chemical inactivation of crude RHDV preparation derived from the livers of infected rabbits. Inactivated vaccines are effective for controlling RHD, but the potential problems of biosafety and animal welfare have negative effects on the application of inactivated vaccines. In this study, an oral Lactobacillus casei (L. casei) vaccine was used as an antigen delivery system to express RHDV capsid protein VP60(VP1)-eGFP fusion protein. The expression of the recombinant protein was confirmed via western blotting and immunofluorescence (IFA). Our results indicate that oral administration of this probiotic vaccine can stimulate secretory immunoglobulin A (SIgA)-based mucosal and IgG-based humoral immune responses in rabbits. The immunized rabbits were completely protected against challenge with RHDV. Our findings indicate that the L. casei expression system is a new strategy for the development of a safe and efficient vaccine against RHDV.

Oral administration of Lactobacillus casei Shirota improves recovery of hand functions after distal radius fracture among elder patients: a placebo-controlled, double-blind, and randomized trial

BackgroundTo evaluate the effect of oral Lactobacillus casei Shirota (LcS) administration on recovery of hand functions in senior patients diagnosed with an acute distal radius fracture.MethodsThis clinical trial is double-blind and placebo-controlled, in which 293 senior patients with distal radius fracture were initially enrolled. After exclusion, 264 eligible patients were randomly assigned to receive oral placebo or LcS daily for a period of 3 months after the fracture. Treatment outcomes were Michigan Hand Questionnaire (MHQ) score, radial deviation and inclination, and ulnar deviation and variance, all of which were monitored and measured every month.ResultsThroughout the length of this study, MHQ score, radial deviation and inclination, and ulnar deviation and variance of patients on oral LcS displayed a significantly faster improvement in comparison to those receiving placebo, over the 3-month intervention period.ConclusionOral administration of LcS dramatically accelerated hand function recovery in senior patients with distal radius fracture.

Liposomes of probiotic's lyophilized cell free supernatant; a potential cosmeceutical product.

The potential uses of lyophilized cell free supernatant (CFS) of human oral derived Lactobacillus paracasei SD1 and Lactobacillus rhamnosus SD11 as cosmeceutical ingredients were investigated in the present study. Lyophilized CFS of both strains showed the antioxidant activity in concentration dependent manner. They also exhibited antimicrobial activity against P. acne, S. aureus and S. epidermidis. In combination, these two strains produced synergistic responses, not only on antioxidant activity but also on antimicrobial activity. A liposomal delivery system was employed to mask the unpleasant colour and odour of CFS. The optimal liposome formulation was characterized by a particle size of 344 nm, PDI of 0.19, zeta value of -48.05 mV and %EE of 69.45. The cytotoxicity results showed that the lyophilized CFS, which was toxic, became non-toxic after encapsulating into liposomes. Altogether, current findings demonstrate the worthiness of development of liposomes of probiotic's lyophilized CFS for cosmeceutical applications.

Influence of Medicinal Plant Extracts on the Growth of Oral Pathogens Streptococcus Mutans and Lactobacillus Acidophilus: An In-Vitro Study.

AIM:This study investigated the antibacterial efficacy of five plant extracts, as well as the combinations of the two most effective plant, extracts either with or without commercial varnish (MI varnish) on the in vitro growth of Streptococcus mutans and Lactobacillus acidophilus in comparison to MI varnish using agar disk diffusion and broth dilution methods.METHODS:Methanolic extractions of five plants (Cinnamon, Turmeric, Ginger, Clove and Black seed,) were tested against the growth of the two oral pathogens. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for the two most effective extracts, and their combinations with different ratios were evaluated against the growth of the two oral pathogens, followed by incorporating the two effective plants or each into commercial MI varnish to be assessed against the oral pathogens in comparison to MI varnish.RESULTS:Only Cinnamon and Clove produced inhibition zones against Streptococcus mutans and Lactobacillus acidophilus growth. MIC for the two plants showed equal antimicrobial activity against Streptococcus mutans, while Cinnamon had a higher sensitivity to Lactobacillus acidophilus than Clove. A mixture of Cinnamon and Clove in a ratio 1:2 exhibited the highest antibacterial activity. Integration the mixture of both plants into MI varnish in a ratio of 1:1:1 presented the highest antibacterial activity. Meanwhile, the lowest one was recorded for the MI varnish alone.CONCLUSION:Methanolic extract of Cinnamon and Clove has considerable antimicrobial activity against Streptococcus mutans and Lactobacillus acidophilus and a new tool for minimally invasive and adhesive dentistry avenues.

Efecto inhibidor de Lactobacillus aislado de la cavidad oral contra patógenos bacterianos y su efecto en la promoción de la salud

Objetivos: Determinar el efecto inhibidor del Lactobacillus aislado de la cavidad oral contra patógenos bacterianos e investigar la correlación entre la presencia de cepas de Lactobacillus y la promoción de la salud. Método: se recolectaron cien muestras de saliva de la cavidad oral de consumidores de productos lácteos y se investigó el aislamiento e identificación de la cepa de Lactobacillus mediante cultivo convencional y secuenciación de 16SrRNA. Además, se realizó un ensayo de difusión en pocillos para determinar la actividad antibacteriana de las cepas de Lactobacillus contra patógenos bacterianos que incluyen Salmonella typhimurium, Klebsiella pneumoniae, Shigella sonnei, Shigella dysenteriae, Enterococcus faecalis y Enterococcus faecium. Finalmente, se investigó la asociación entre el estado de salud y el aislamiento de Lactobacillus y se obtuvieron los datos utilizando el cuestionario mediante la prueba de chi-cuadrado. Resultados: Se aislaron treinta cepas de Lactobacillus de 100 muestras de saliva. La cepa aislada más común fue L. gasseri (n = 18), seguida por L. vaginalis (n = 3) y L. salivarius (n = 3). Todas las cepas de Lactobacillus demostraron actividad antibacteriana contra al menos uno de los patógenos investigados. Sin embargo, los resultados más fuertes fueron obtenidos por L. vaginalis contra K. pneumonia. No se encontraron correlación entre la presencia de algunas de las treinta cepas de Lactobacillus y la promoción de la salud. Sin embargo, solo la especie L. gasseri tuvo un impacto positivo significativo en la salud de sus hospedadores (P &lt;0,05). Conclusión: solo algunas especies de Lactobacillus tienen un impacto positivo en la promoción de la salud. A pesar de la actividad débil contra los patógenos investigados, L. gasseri tiene un impacto positivo en el problema mental (ira intensa y depresión) de sus huéspedes.

Beneficial Effects of Oral Lactobacillus on Pain Severity in Women Suffering from Endometriosis: A Pilot Placebo-Controlled Randomized Clinical Trial.

BackgroundThis study assessed the effects of a lactobacillus-based medication on pain intensity scores in women with endometriosis.Materials and MethodsThe present randomized pilot placebo-controlled trial was done on eligible women who were surgically and pathologically diagnosed with endometriosis. Thirty-seven participants who had not received hormonal treatment in the last three months, were enrolled and randomized into LactoFem®and placebo groups. Lactobacillus capsules or placebo were administrated orally once a day for 8 weeks. Patients were assessed for pain severity using Visual Analogue Scale (VAS) scores for dysmenorrhea, dyspareunia and chronic pelvic pain at baseline and after 8 and 12 weeks post-intervention.ResultsMean age of participants and mean body mass index (BMI) for the LactoFem®and control groups were compara- ble. All patients had stage 3 and 4 of the disease based on revised American fertility society (AFS) classification of endome- triosis. Mean initial pain scores for dysmenorrhea, dyspareunia and chronic pelvic pain were 6.53 ± 2.88, 4.82 ± 3.76 and 4.19 ± 3.53, respectively in the LactoFem®group and 5.60 ± 2.06, 3.67 ± 2.64 and 2.88 ± 2.80, respectively for the control group; the two groups had comparable scores in this regard. There was more decrease in pain scores for both dysmenorrhea and the overall pain after 8 weeks of treatment in LactoFem®group compared to the control group. The scores for dysmen- orrhea were 6.53 ± 2.88 and 5.60 ± 2.06 in the LactoFem®and control groups, respectively, before intervention but, after 8-week treatment, these values were 3.07 ± 2.49 and 4.47 ± 2.13 (P=0.018), respectively. The changes in overall pain score in the LactoFem® and control group during this period were 7.33 ± 7.00 and 4.11 ± 1.68, respectively (P=0.017).ConclusionThis study showed some beneficial effects of lactobacillus administration on endometriosis-related pain (Registration number: IRCT20150819023684N5).

Oligosaccharides as co-encapsulating agents: effect on oral Lactobacillus fermentum survival in a simulated gastrointestinal tract.

Four kinds of oligosaccharides were used as co-encapsulating agents to test the effect of extrusion-based microencapsulation on protection of Lactobacillus fermentum L7 against exposure to simulated gastric and intestinal juices as well as long-term refrigeration storage at 4°C. The combination of alginate with galacto-oligosaccharides, isomalto-oligosaccharides, fructo-oligosaccharides, and xylo-oligosaccharides, or alginate alone exhibited good properties of the beads. The diameters of the cell beads co-encapsulated with oligosaccharides and encapsulated with alginate alone were similar, in the range of 2.34-2.51mm. However, the encapsulation yield of L. fermentum cells co-encapsulated with oligosaccharides, which was in the range of 79.52-89.75%, was significantly higher than that of cells encapsulated with alginate alone. The capsules were stable in gastric conditions and can disintegrated when exposed to intestinal conditions. Additionally, the viability of microencapsulated cells after exposure to the simulated gastric and intestinal juices as well as long-term refrigeration storage was better than that of free cells, and the viability of cells co-encapsulated with oligosaccharides was better than that of cells encapsulated with alginate alone. Furthermore, fructo-oligosaccharides used as co-encapsulating agent showed the best performance. Microencapsulating L. fermentum with oligosaccharides protected cells well at a low temperature and offered effective gastrointestinal delivery of probiotics, and thus has the potential to maintain bacterial survival in probiotic products and will provide the research basis for design of effective probiotic-prebiotic combinations to maximize host benefit.

Effectiveness of Oral Probiotics as Adjuvant Therapy in Reproductive Aged Women with Vaginal Discharge

&#x0D; Objective: To investigate the efficacy of oral proboiotics and prove the high proportion of cure and satisfaction levels of post-treatment patients with a combination of antimicrobial-probiotic oral Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 compared to a combination of antimicrobial-placebo in the treatment of reproductive aged patients with vaginal discharge in the outpatient obstetrics and gynecologic clinic in Dr. Cipto Mangunkusumo Hospital, Jakarta, Indonesia and Arifin Achmad Regional Hospital Pekanbaru, Riau, Indonesia.&#x0D; Methods:This was a randomized, double-blind, placebo-controlled trial involving50 subjects consisting of reproductive aged women. Data were collected using syndromic approach, probiotics were given as an adjuvant for standard antimicrobial therapy versus placebo as control, response was recorded 4 weeks later, for cure proportion and satisfaction level. Statistical analysis was performed to assess the variables. Interim analysis with conditional power assesment and futility testing were performed at midway due to insufficient sample size. Research was approved by Ethics Commitee for Health Researches Faculty of Medicine University of Indonesia-Dr. Cipto Mangunkusumo Hospital in March 2016.&#x0D; Results: A total of 50 subjects participated in this study. and analyzed (25 subjects in each group), cure proportion 56%(14) of the treatment and 60%(15)on the control group, with relative risk of 1.1, Chi-square test p value (0.77, 95% CI; 0.57 to 2 , 11). High satisfaction level (score ≥67) was higher in the placebo (52,6%, 10 subjects) compared to probiotic group (47,4%, 9 subjects), p value 0,65 (≥0,05). Conditional power and futility testing curve, revealed Z = -0.2865, conditional power 0.11 to 0.13, and futility index of 0.87 to 0.88, equals to low possibility of statistical significance with full sample size (84).&#x0D; Conclusion: There was no clinical and statistical difference in the proportion of cure rate and the level of satisfaction in patients of probiotics vs placebo groups after treatment for 4 weeks. The initial hypothesis of higher proportion of the cure ratein the treatment group still cannot be excluded, due to insufficient samples.&#x0D; Keywords: bacterial vaginosis, , lactobacillus reuteri RC-14, lactobacillus rhamnosus GR-1, randomized double blind controlled trial, trichomoniasis , vaginal discharge, vulvovaginal candidiasis.

An orally administered oral pathobiont and commensal have comparable and innocuous systemic effects in germ-free mice.

We recently proposed a novel mechanism linking periodontitis and systemic diseases, in which orally administered Porphyromonas gingivalis affects gut microbiota composition and subsequently leads to systemic inflammation. However, the mechanism by which P.gingivalis generates systemic effects from the gut is unknown. Six-week-old germ-free mice were orally administered with either an oral pathobiont P.gingivalis or an oral commensal Lactobacillus salivarius twice a week for 5weeks. Control mice were administered with vehicle only. Alveolar bone resorption was evaluated histologically. The expression profile of various genes was analyzed in gingival tissue, liver, small intestine and large intestine using real-time polymerase chain reaction. Sera were analyzed for antibody, endotoxin and interleukin (IL)-6 levels. Antibody levels were also analyzed for culture supernatant of cells from mesenteric lymph nodes and spleens. A proportion of T-helper 17 and Treg in the cells from mesenteric lymph nodes and spleens was analyzed by flow cytometry. The level of IL-6 and IL-17 in the cell culture supernatants was analyzed by enzyme-linked immunosorbent assay. P.gingivalis administration did not induce alveolar bone resorption. Although P.gingivalis elicited systemic antibody response in germ-free mice, unlike in specific pathogen-free mice, P.gingivalis did not induce an inflammatory response in gingiva, liver and intestinal tissue, or alter the proportion of T-helper 17 and Treg. However, IL-6 and IL-17 productions were significantly elevated and tended to be elevated, respectively, in the cells from mesenteric lymph nodes of P.gingivalis-administered mice. Interestingly, the expression of IL-10 and tight junction protein in the gingiva and intestine, respectively, was significantly upregulated in P.gingivalis-treated mice. Administration of L.salivarius elicited almost similar effects as P.gingivalis. The oral pathobiont P.gingivalis did not induce any detectable pathogenic changes or any major host responses when administered to germ-free mice. There may be indirect mechanisms for gut-mediated systemic effects by P.gingivalis.

New Polyhydroxylated Steroidal Saponins from Solanum paniculatum L. Leaf Alcohol Tincture with Antibacterial Activity against Oral Pathogens.

Solanum paniculatum L. is widely used in Brazilian folk medicine for the treatment of liver and gastrointestinal disorders as well as for culinary purposes and beverage production. Fractionation of hydroalcoholic [ethanol (EtOH) 70%] tincture from S. paniculatum leaves led to the isolation of six new spirostanic saponins which included 6- O-α-l-rhamnopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 R,25 S)-3β,6α,23-trihydroxy-5α-spirostane (1), 6- O-β-d-xylopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 R,25 R)-3β,6α,23-trihydroxy-5α-spirostane (4), 3- O-α-l-rhamnopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 S,25 R)-3β,6α,23-trihydroxy-5α-spirostane (5), 3- O-β-d-xylopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,23 S,25 R)-3β,6α,23-trihydroxy-5α-spirostane (6), 6- O-α-l-rhamnopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,25 S)-1β,3β,6α-trihydroxy-5α-spirostane (7), and 6- O-β-d-xylopyranosyl-(1''→3')-β-d-quinovopyranosyl-(22 S,25 S)-3β,4β,6α-trihydroxy-5α-spirostane (8) together with two known spirostanic saponins (2, 3). The structures of these compounds were determined by one-dimensional (1D) and two-dimensional (2D) NMR experiments in addition to high-resolution electrospray ionization mass spectrometry (HRESIMS) analyses. The 70% alcohol tincture, used as phytomedicine, exhibited promising activities against oral pathogens, including, Steptococcus sanguinis, St. oralis, St. mutans, St. mitis, and Lactobacillus casei with minimal inhibitory concentration (MIC) values ranging from 6.25 to 50 μg/mL. The saponin fraction, nonetheless, showed lower activity against all the strains tested (from 100 to >400 μg/mL).

Bio-therapeutic potential of lactic acid bacterial against &lt;i&gt;Aeromonas&lt;/i&gt; species

Among the microorganisms responsible for human deaths from gastroenteritis, little or no attention is given to Aeromonas species. Due to wrong diagnosis and virulence properties exhibited by Aeromonads including drug resistant profile, these organisms have constituted challenges in public health, which may however, needed alternative control strategy. Lactic acid bacteria (LAB) are generally regarded as safe and confer health benefit in humans due to antimicrobial compounds produced and could be used to control infections caused by Aeromonas species. Aeromonas species and LAB were characterized using 16S rRNA. Bacteriocin production and antagonistic activity of LAB metabolites against Aeromonas species were determined using agar well diffusion and the best LAB selected based on the above parameters for in vivo tests. The effect of the selected LAB on Wistar rat infected with selected Aeromonas species based on virulence properties produced was determined through haematological, serum enzymes (alanine amino tranferase (ALT), alkaline phosphatase (ALP) and aspartate amino transferases (AST) and histopathology analysis. Data were analysed using descriptive statistics and ANOVA at α 0.05 . The 16S rRNA confirmed Aeromonas species as A. hydrophilla A. caviae, A. veronii, A. simiae , A. punctata, A. finlandiensis , Plesiomonas shigelloides and LAB as Lactobacillus plantarum (LAB 01) , Lactobacillus paraplantarum (LAB02), Lactobacillus fermentum (LAB3) and Lactobacillus oris (LAB 04). Antagonistic activity of both the LAB metabolites and bacteriocin showed inhibition zones ranging from 18-31mm against Aeromonas species. Lactobacillus plantarum (LAB01) produced 90.15 mg/100g of lactic acid. Wistar rats infected with A. hydrophila (ST08) and administered with L. plantarum (LAB01) showed no significant differences from the control rats in terms of haematology, enzymatic and histopathology parameters. However, Wister rats infected with A. hydrophila (ST07) only had 17.47±0.01x10 9 /L white blood cells compared to the control (2.31 ± 0.14x10 9 /L). Enzymatic values revealed high ALT (34.13 ± 0.01IU/L), AST (37.11 ± 0.00IU/L) and ALP (32.13 ± 0.02IU/L) against control (16.03 ± 0.03 - 24.10 ± 0.01IU/L). Histopathology analysis revealed renal cortex showing global necrosis in infected rats. Howe Lactobacillus plantrum (LAB01) produced antimicrobial compounds that inhibited multiple antibiotic resistant Aeromonas species in vitro and in vivo and could be used as bio-therapeutic agents against Aeromonads infection. Keywords : Aeromonas species, lactic acid bacteria, Wister rats, bacteriocin, organic acid

Oral administration of Lactobacillus casei Shirota can ameliorate the adverse effect of an acute aflatoxin exposure in Sprague Dawley rats.

Aflatoxin B1(AFB1) is a toxic compound commonly found in some crops with an adverse health effect on human and animals. Some beneficial microorganisms (or probiotics) such as lactic acid bacteria have shown the ability to reduce the bioavailability of aflatoxins and its intestinal absorption. However, the dose and duration of aflatoxins exposure and probiotic treatment can influence the ability of probiotics to remove aflatoxins. Therefore, this research aimed to investigate the efficacy of oral probiotic Lactobacillus casei Shirota strain (LcS) induction in an acute exposure to AFB1 in rats. Experimentally, Sprague Dawley rats were divided into three groups: AFB1 only (n = 9); AFB1 treated with LcS (n = 9); and control (no AFB1 exposure) (n = 6) groups. The blood AFB1 level of rats treated with LcS was slightly lower than the untreated AFB1 induced rats (11.12 ± 0.71 vs 10.93 ± 0.69 ng g-1). Also, LcS treatment slightly moderated the liver and kidney biomarkers in AFB1 induced rats. However, a trend for a significant difference was only observed in ALT of AFB1 induced rats treated with LcS compared to their counterparts (126.11 ± 36.90 vs 157.36 ± 15.46, p = 0.06). Rats' body weight decreased in all animals force-fed with AFB1 with no significant difference between LcS treatment compared to the counterpart. In conclusion, this experiment indicated that probiotic LsC was able to slightly ameliorate the adverse effect of an acute exposure to AFB1 in rats. However, future studies with longer probiotics treatment or higher probiotics dose is required to confirm these findings.