The study conducted in Vietnam investigated the effects of the probiotic Lactobacillus casei strain Shirota (LcS) on immune profiles and intestinal microbial translocation among children infected with human immunodeficiency virus (HIV). This prospective study included 60 HIV-infected children-including 31 HIV positive children without antiretroviral therapy and 29 HIV positive children who received antiretroviral therapy for a period of 3.5 years and 20 children without HIV infection . All children were given fermented milk containing LcS (6.5 × 10⁸ cfu) daily for 8 weeks. Before and after LcS ingestion, blood samples were collected for virological, immunological, and bacteriological analyses.
After LcS ingestion, peripheral CD4⁺ T-cell and specific Th2 counts significantly increased in both HIV-infected groups; Th17 counts increased in all three groups; regulatory T-cell counts decreased in the ART(+) and HIV(-) groups; activated CD8⁺ cells decreased from 27.5% to 13.2% (p < 0.001) in HIV positive children; and plasma HIV load decreased slightly but significantly among HIV(+) children. No serious adverse events occurred. These findings suggest that short-term LcS ingestion is a safe supportive approach with immunological and virological benefits in HIV-infected children.
Aim: To determine the immune response of healthy adults to the ingested Probiotic.
Methods: Daily ingestion of a probiotic drink containing Lactobacillus casei strain Shirota (LcS; 1.3 x 1010 live cells) by healthy adults for (i) 4-weeks LcS, (ii) 6-weeks discontinuation of LcS, and (iii) final 4-weeks of LcS, was investigated.
Results: There was a significant increase in expression of the T-cell activation marker CD3+ CD69+ in ex vivo unstimulated blood cells at weeks 10 and 14 and a significant increase in the Natural Killer (NK) cell marker CD3+ CD16/56+ in ex vivo unstimulated blood cells at weeks 4, 10 and 14. Expression of the NK cell activation marker CD16/56+ CD69+ in ex vivo unstimulated blood cells was 62% higher at week 10 and 74% higher at week 14. Intracellular staining of IL-4 in ex vivo unstimulated and PMA/ionomycin-stimulated CD3+ β7+ integrin blood cells was significantly lower at week 10 and 14. Intracellular staining of IL-12 in ex vivo unstimulated and LPS-stimulated CD14+ blood cells was significantly lower at weeks 4, 10 and 14. Intracellular staining of TNF-α in LPS-stimulated CD14+ blood cells was significantly lower at weeks 4, 10 and 14. Mucosal salivary IFN-γ, IgA1 and IgA2 concentrations were significantly higher at week 14 but LcS did not affect systemic circulating influenza A-specific IgA or IgG and tetanus specific IgG antibody levels.
Conclusions: In addition to the decrease in CD3+ β7+ integrin cell IL-4 and a CD14+ cell anti-inflammatory cytokine profile, at week 14 increased expression of activation markers on circulating T cells and NK cells and higher mucosal salivary IgA1 and IgA2 concentration indicated a secondary boosting effect of LcS.
Aim: To evaluate the effect of Lactobacillus casei strain Shirota-fermented milk (LcS-FM) on the incidence of Upper Respiratory Tract Infections (URTI) in healthy idle aged office workers.
Methods: Study was conducted in 96 healthy middle-aged office workers in the age group of 30 – 49 years. The office workers consumed LcS-Fermented Milk containing 1.0 × 1011 LcS bacteria or control milk (CM) once daily for 12 weeks during the winter season. URTI episodes were evaluated by a physician via a questionnaire of URTI symptoms.
Results: The results of the study showed that the incidence of URTI’s during the intervention period were significantly lower in the LcS-FM group as compared to the CM group (22.4 vs. 53.2 %, P = 0.002). The time to-event analysis showed that the LcS-FM group had a significantly higher URTI-free rate than the CM group over the test period (log-rank test: χ2 11.25, P = 0.0008). The cumulative number of URTI episodes and cumulative days with URTI symptoms per person was lower in the LcS-FM group, and the duration per episode was also shorter. There was no reduction in NK cell activity in peripheral blood mononuclear cells and no increase in salivary cortisol levels in the LcS-FM group.
Conclusions: The conclusion of the study suggests that the daily intake of fermented milk with LcS may reduce the risk of URTI’s in healthy middle-aged office workers, probably through modulation of the immune system.
Aim: The efficacy of probiotic Lactobacillus casei (Shirota strain) in reducing incidence of ventilator-associated pneumonia (VAP) in medical patients who received mechanical ventilation at Siriraj Hospital was evaluated.
Method: A prospective, randomized, open-label controlled trial was conducted on 150 adult hospitalized patients in medical wards who were on mechanical ventilation for 72 hours or longer: The patients were randomized to the probiotic group or the control group. The patients in the probiotic group received 80 ml of a probiotic fermented milk drink containing 8 billion Lactobacillus casei (Shirota strain) for oral care after having standard oral care once daily and additional 80 ml of the aforementioned fermented dairy product was given via enteral feeding once daily. The primary outcomes were incidence of VAP and incidence rate of VAP episodes per 1,000 ventilator-days. The secondary outcomes were length of hospital stay, mortality at day 28 and 90, incidence of diarrhea, and presence of resistant bacteria in oropharyngeal and rectal swab samples taken from the patients at baseline, day 7 and day 28 after enrollment.
Results: The baseline characteristics of the patients in the probiotic group (75) and the control group (75) were not significantly different. The patients in the probiotic group were less likely to develop VAP compared with the control group (24% vs. 29.3%, p = 0.46). The incidence rates of VAP in the probiotic and control groups were 22.64 and 30.22 episodes per 1,000 ventilator-days, respectively (p = 0.37). A trend of lower prevalence of resistant bacteria cultured from oropharyngeal swabs in the probiotic group than that in the control group was observed. Overall 28 and 90-day mortality and length of hospital stay of the patients in both groups was not significantly different.
Conclusions: Administration of probiotic containing Lactobacillus casei (Shirota strain) resulted in a tendency to reduce the incidence of VAP and colonization with resistant bacteria in oropharyngeal cavity without significant effects on mortality and length of hospital stay.
Intestinal microbiota profiles of healthy pre-school and school-age children and effects of probiotic supplementation.Wang C et al 2015, Ann Nutr Metab 67:257-266.
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Aim: To demonstrate the gastrointestinal survival of Lactobacillus casei strain Shirota (LcS) in healthy Vietnamese adults.
Methods: Twenty-six healthy Vietnamese adults took part in the study. Each participant consumed 65 mL of a fermented milk drink containing LcS daily for 14 days. The drink contained a dose of 10 8 CFU/mL LcS. Fecal samples were collected before, during and after consuming the fermented milk drink.
Results: LcS was confirmed by culture and ELISA. After 7 and 14 days of ingesting fermented milk drink, LcS was recovered from fecal samples at average of 5.0×10 7 CFU/g feces (n=26) and 5.4×10 7 CFU/g feces (n=26), respectively. LcS persisted in 8 volunteers until day 42 (after 14 days stopping fermented milk drink) at 0.0033×10 7 CFU/g feces (n=8).
Conclusions: Confirmed the survival of LcS after passage through the gastrointestinal tract of Vietnamese adults.
Aim: Although several studies have demonstrated the efficacy of probiotics for preventing upper respiratory tract infections (URTIs) in at-risk populations, including children and the elderly. Few studies have investigated the efficacy of probiotics in healthy adults living normal, everyday lives. Thus, this study evaluated the effect of Lactobacillus casei strain Shirota-fermented milk (LcS-FM) on the incidence of URTIs in healthy middle-aged office workers.
Method: In a randomized controlled trial, 96 male workers aged 30–49 years consumed LcS-FM containing 1.0 × 1011 LcS bacteria or control milk (CM) once daily for 12 weeks during the winter season. URTI episodes were evaluated by a physician via a questionnaire of URTI symptoms.
Results: The incidence of URTIs during the intervention period was significantly lower in the LcS-FM group as compared to the CM group (22.4 vs. 53.2 %, P = 0.002). The time to-event analysis showed that the LcS-FM group had a significantly higher URTI-free rate than the CM group over the test period (log-rank test: χ2 11.25, P = 0.0008). The cumulative number of URTI episodes and cumulative days with URTI symptoms per person was lower in the LcS-FM group, and the duration per episode was shorter. Inhibition of both reductions in NK cell activity in peripheral blood mononuclear cells and increase in salivary cortisol levels was observed in the LcS-FM group.
Conclusions: The results suggest that the daily intake of fermented milk with LcS may reduce the risk of URTIs in healthy middle-aged office workers, probably through modulation of the immune system.
Aim: There is insufficient evidence of preventive effect of probiotics on upper respiratory tract infections (URTIs) in an elderly population.
Method: A multicenter, double-blinded, randomized, placebo-controlled parallel group study was conducted. Elderly persons had participated who used day care at 4 facilities in Tokyo. We used fermented milks containing Lactobacillus casei strain Shirota (LcS) and placebo drinks as test drinks.
Results: A total of 154 subjects was analyzed. The number of persons diagnosed with an acute URTIs was almost identical in both groups (LcS: 31, placebo: 32), whereas the number of acute URTIs events (LcS: 68, placebo: 51) and the symptom score (LcS: 425, placebo: 396) were both higher in the LcS group. Permutation tests performed using the total number of acute URTIs infection events/total days of observation and the total symptom score/total days of observation found no statistically significant difference respectively (P values of .89 and .64, respectively). Comparing the mean duration of infection per infection event found a shorter mean duration in the LcS group (LcS: 3.71 days, placebo: 5.40 days), and the difference was statistically significant.
Conclusions: The results suggest that fermented milk containing LcS probably reduces the duration of acute URTIs.
The use of probiotics as dietary approach to prevent exposure to food contaminant, aflatoxin B1 (AFB1) has greatly increased. Several studies found that AFB1 binding to the bacterial cell wall is strain-specific. Moreover, the interaction between AFB1 and bacterial cell wall is not well-understood, thus warrants further investigation. This research was conducted to assess the ability of Lactobacillus casei Shirota (LcS) to bind AFB1 at different concentrations and to determine AFB1 binding efficiency of different LcS cell components including live cell, heat-treated, and cell wall. In addition, the interaction between AFB1 and LcS was also evaluated via scanning electron microscopy (SEM) and through an animal study. The binding of AFB1 by all LcS cell components depends on the concentration of available AFB1. Among all LcS cell components, the live LcS cells exhibited the highest binding efficiency (98%) toward AFB1. Besides, the SEM micrographs showed that AFB1 induced structural changes on the bacterial cell surface and morphology including rough and irregular surface along with a curve rod-shaped. In vivo experiment revealed that LcS is capable of neutralizing the toxicity of AFB1 on body weight and intestine through the binding process. The animal's growth was stunted due to AFB1 exposure, however, such effect was significantly (p < 0.05) alleviated by LcS. This phenomenon can be explained by a significant (p < 0.05) decreased level of blood serum AFB1 by LcS (49.6 ± 8.05 ng/mL) compared to AFB1-exposed rats without treatment (88.12 ± 10.65 ng/mL). Taken together, this study highlights the potential use of LcS as a preventive agent against aflatoxicosis via its strong binding capability.
Considerate proportion of elderly patients with a rib fracture are susceptible to pulmonary complications, especially pneumonia. This study aimed to assess the effect of oral administration of the probioticLactobacillus casei Shirota(LcS) on pneumonia and pulmonary functions among elderly patients with single rib fracture. The current study included 204 eligible elderly patients with a single rib fracture. Patients were randomly assigned to receive oral administration of skimmed milk containing either a commercial probiotic LcS or placebo daily for 1 month after the fracture. This was followed by pneumonia assessments, pulmonary function testing including forced expiratory volume (FEV), negative inspiratory pressure (NIP), and forced vital capacity (FVC), as well as evaluation of potential adverse effects including myocardial infarction, acute kidney injury, nonunion of fractured bone, or stroke. After 1-month consumption, patients in the LcS group exhibited decreased pneumonia and increased recovery of pulmonary functions, in terms of FEV, FVC, and NIP, compared to the placebo group. No difference was observed in incidence of adverse events between the 2 groups. In patients with a single rib fracture, oral administration of the probiotic LcS was associated with a lower incidence of pneumonia and increased pulmonary functions without causing severe adverse effects.
Suppression of immune function during long spaceflights is an issue that needs to be overcome. The probiotic Lactobacillus casei strain Shirota (LcS) could be a promising countermeasure, and a project was launched to investigate the efficacy of its use on the International Space Station (ISS). As a first step, a special probiotic product for space experiments, containing freeze-dried LcS in capsule form (probiotics package) was developed. This was for tested its stability through 1 month of storage on the ISS. The temperature inside the ISS ranged from 20.0 to 24.5 °C. The absorbed dose rate of the flight sample was 0.26 mGy/day and the dose equivalent rate was 0.52 mSv/day. The number of live LcS was 1.05 × 1011 colony-forming units/g powder (49.5% of the initial value) 6 months after the start of the study; this value was comparable to those in the two ground controls. Profiles of randomly amplified polymorphic DNA, sequence variant frequency, carbohydrate fermentation, reactivity to LcS-specific antibody, and the cytokine-inducing ability of LcS in the flight sample did not differ from those of the ground controls.
Evidence suggests that the gut microbiota might play an important role in fibromyalgia syndrome (FMS) and chronic fatigue syndrome (CFS). The goal of the study was to systematically review the reported effect of probiotic treatments in patients diagnosed with FMS or CFS. A systematic review was carried out using 14 databases (PubMed, Cochrane Library, Scopus, PsycINFO, and others) in February 2016 to search for randomised controlled trials (RCTs) and pilot studies of CFS or FMS patient, published in the last ten years (from 2006 to 2016). The Jadad scale was used to asseverate the quality of the clinical trials considered. Two studies (n=83) met the inclusion criteria, which were performed in CFS patients and both studies were considered as a 'High range of quality score'. The administration of Lactobacillus casei strain Shirota in CFS patients, over the course of 8 weeks, reduced anxiety scores. Likewise, this probiotic changed the faecal composition following 8 weeks of treatment. Additionally, the treatment with Bifidobacterium infantis 35624 in CFS patients, during the same period, reduced inflammatory biomarkers. The evidence about the usefulness of probiotics in CFS and FMS patients remains limited. The studied strains of probiotics have demonstrated a significant effect on modulating the anxiety and inflammatory processes in CFS patients. However, more experimental research, focusing mainly on the symptoms of the pathologies studied, is needed.
The study aimed to examine whether Lactobacillus casei strain Shirota (LcS) improves sleep quality under psychological stress. A double-blind, placebo-controlled trial was conducted in healthy fourth year medical students exposed to academic examination stress. The trial was repeated over two consecutive years in different groups of students, and the data were pooled. For 8 weeks prior to and 3 weeks after a National Standardized Examination, a total of 48 and 46 subjects received a daily dose of 100 ml of LcS-fermented milk or non-fermented placebo milk, respectively. The study measured subjective anxiety, overnight single-channel electroencephalography (EEG) recordings, and the Oguri-Shirakawa-Azumi (OSA) sleep inventory scores of subjective sleeps quality. Total OSA scores were significantly lower than baseline on the day before the exam and recovered after the exam, indicating a stress-induced decline in sleep quality. There was a significant positive effect of LcS treatment on OSA factors for sleepiness on rising and sleep length. Sleep latency measured by EEG lengthened as the exam approached in the placebo group but was significantly suppressed in the LcS group. The percentage of stage 3 non-REM (N3) sleep decreased in the placebo group as the exam approached, whereas it was maintained in the LcS group throughout the trial. Delta power during the first sleep cycle, measured as an index of sleep intensity, increased as the exam approached in the LcS group and was significantly higher than in the placebo group. The study suggests that daily consumption of LcS may help to maintain sleep quality during a period of increasing stress. The observed retention of N3 sleep and increased delta power in the LcS group may have contributed to higher perceived sleep satisfaction.
The study was conducted to investigate the effects of the probiotic Lactobacillus casei strain Shirota (LcS) on abdominal dysfunction, a double-blind, placebo-controlled trial with healthy medical students undertaking an authorized Nationwide examination for academic advancement. 23 and 24 subjects consumed an L. casei strain Shirota-fermented milk and a placebo milk daily, respectively for 8 weeks, until the day before the examination. In addition to assessments of abdominal symptoms, psychophysical state, and salivary stress markers, gene expression changes in peripheral blood leukocytes and composition of the gut microbiota were analyzed using DNA microarray analysis and 16S rRNA gene amplicon sequence analysis, respectively, before and after the intervention. Stress-induced increases in a visual analog scale measuring feeling of stress, the total score of abdominal dysfunction, and the number of genes with changes in expression of more than 2-fold in leukocytes were significantly suppressed in the L. casei strain Shirota group compared with those in the placebo group. A significant increase in salivary cortisol levels before the examination was observed only in the placebo group. The administration of L. casei strain Shirota, but not placebo, significantly reduced gastrointestinal symptoms. Moreover, 16S rRNA gene amplicon sequencing demonstrated that the L. casei strain Shirota group had significantly higher numbers of species, a marker of the alpha-diversity index, in their gut microbiota and a significantly lower percentage of Bacteroidaceae than the placebo group.The study demonstrated that the daily consumption of the probiotic strain L. casei strain Shirota, preserves the diversity of the gut microbiota and may relieve stress-associated responses of abdominal dysfunction in healthy subjects exposed to stressful situations.
Aim: This study investigated relationships between the frequent intake of fermented milk products containing Lactobacillus casei strain Shirota (LcS) and the onset of hypertension (resting systemic pressure ≥140 mmHg [systolic]/≥90 mmHg [diastolic].
Method: A 5-year period of study was conducted in 352 Japanese who were in the age group of 65 to 93 years, (125 men and 227 women). The subjects were divided into two groups (n=254 and n=98) on the basis of their intake of fermented milk products (less than 3 times a week or more than 3 times a week).
Results: The incidence of hypertension over the 5-year interval was significantly lower in those who took fermented milk products more than 3 times rather than less than 3 times/week (6.1 vs 14.2%, P=0.037). A multivariate-adjusted proportional hazards model predicted that blood pressures were significantly more likely to remain normal over 5 years in subjects who took more than 3 times fermented milk products rather than less than 3 times/ week (relative risk 0.398 [95% confidence interval 0.167-0.948], P=0.037).
Conclusions: These results suggest that after adjustment for potential confounders, the risk of developing hypertension is substantially lower in elderly people who take fermented milk products containing LcS at least 3 times a week.
Aim: To clarify the usefulness of Lactobacillus casei strain Shirota (LcS) - fermented milk in the normalization of bowel movements and improvement of infection control for the elderly residents and staff of facilities for the elderly.
Method: The study was performed on elderly residents (average age, 85) and staff members (average age, 37) of facilities for the elderly. Participants were divided into two groups based on their intake of Lactobacillus casei strain Shirota (LcS) fermented milk or a placebo once daily for 6 months.
Results: A significantly lower incidence of fever and improved bowel movement was seen in the LcS - fermented milk group (n=36) in comparison to the placebo group (n=36). The numbers of Bifidobacterium and Lactobacillus were higher (p < 0.01) whereas pathogenic organisms such as Clostridium difficile were significantly lower (p < 0.05), fecal acetic acid concentration and total acidity was higher in the LcS group. A significant difference in the intestinal microbiota, fecal acetic acid, and pH was also observed between the LcS and placebo groups among the staff members.
Aim: To determine whether a fermented milk drink containing probiotics could improve the bowel habits of frail elderly individuals living in a nursing home.
Method: The bowel habit (stool quality and bowel movements) of 135 participants was recorded by nursing staff during a baseline period of 3 weeks. After this period participants received daily a fermented milk drink containing minimally 6.5×109 colony forming units of Lactobacillus casei Shirota (LcS) for 6 weeks. During this period, bowel habits was recorded and compared to baseline period. 44 participants (74-99 years old) was compliant and used for analysis.
Results: Consumption of fermented milk containing LcS significantly increased the percentage of ideal stool types per week (P<0.01), lowered the percentage of constipation stool types per week (P<0.01) and significantly lowered the percentage of diarrhoea stool types per week (P=0.016) as compared to the baseline period.
Conclusions: The results suggest that a fermented milk containing LcS significantly improves the bowel habits of frail elderly residents in a nursing home. The results of this study need further substantiation.
Conclusions: The study concludes that long-term consumption of LcS -fermented milk may be useful in decreasing the daily risk of infection and improving the quality of life among the residents and staff in facilities for the elderly.
Aim: This pilot study investigated the effects of the probiotic Lactobacillus casei strain Shirota (LcS) on psychological and physical stress responses in medical students undertaking an authorized Nationwide examination for promotion.
Method: In a double-blind, placebo-controlled trial, 24 and 23 healthy medical students consumed a fermented milk containing LcS and a placebo milk, respectively, once a day for 8 weeks until the day before the examination. Psychological state, salivary cortisol, faecal serotonin and plasma L-tryptophan were analysed on 5 different sampling days (8 weeks before examination, 2 weeks before examination, 1 day before examination, immediately after examination and 2 weeks after the examination). Physical symptoms were also recorded in a dairy by subjects during the intervention period for 8 weeks.
Results: In association with a significant elevation of anxiety at 1 day before the examination, salivary cortisol and plasma L-tryptophan levels were significantly increased in only the placebo group (P < 0.05). Two weeks after the examination, the LcS group had significantly higher faecal serotonin levels (P < 0.05) than the placebo group. Moreover, the rate of subjects experiencing common abdominal and cold symptoms and total number of days experiencing these physical symptoms per subject were significantly lower in the LcS group than in the placebo group during the pre-examination period at 5-6 weeks (each P < 0.05) and 7-8 weeks (each P < 0.01) during the intervention period.
Conclusions: The results suggest that daily consumption of fermented milk containing LcS may exert beneficial effect by preventing the onset of physical symptoms in healthy subjects exposed to stressful situations.
14. Decreased duration of acute upper respiratory tract infections with daily intake of fermented milk: A multicenter, double-blinded, randomized comparative study in users of day care facilities for the elderly population.
Fujita R, Iimuro S, Shinozaki T, Sakamaki K, Uemura Y et al (2013) Am J Infect Control pii: S0196-6553(13)00846-8. doi: 10.1016/j.ajic.2013.04.005.