In vitro study of potential prebiotic properties of monovarietal extra virgin olive oils.

Olive oil, essential ingredient of the Mediterranean diet, is attracting a growing interest due to increasing evidence on its beneficial effects on human health. This study investigated whether extra virgin olive oil (EVOO) possess prebiotic properties. Twenty different monovarietal EVOO samples from 5 Marche region cultivars (Italy) were studied. The prebiotic activity of EVOOs was assessed monitoring the selective stimulation of gut bacterial species and the short chain fatty acids (SCFAs) production, using an in vitro fermentation system. All EVOOs selectively stimulated Lactobacillus spp., with a stronger activity than that observed in the inulin fermentation (positive control). Also, the bifidobacteria population increased; this bifidogenic stimulation was of EVOOs from Raggia cultivar. SCFAs appeared significantly higher after 24 h in all EVOO fermentations than in the control. Acetic and propionic acids production was particularly stimulated. Overall, most of the investigated EVOOs had a potential prebiotic activity, similar or stronger than inulin.

Possible Role of Tauroursodeoxycholic Acid (TUDCA) and Antibiotic Administration in Modulating Human Gut Microbiota in Home Enteral Nutrition Therapy for the Elderly: A Case Report.

Tauroursodeoxycholic acid (TUDCA) increases the influx of primary bile acids into the gut. Results obtained on animal models suggested that Firmicutes and Proteobacteria phyla are more resistant to bile acids in rats. As part of a pilot study investigating the role of probiotics supplementation in elderly people with home enteral nutrition (HEN), a case of a 92-year-old woman with HEN is reported in the present study. She lives in a nursing home and suffers from Alzheimer's disease (AD); the patient had been prescribed TUDCA for lithiasis cholangitis. The aim of this case report is therefore to investigate whether long-term TUDCA administration may play a role in altering the patient's gut microbiota (GM) and the impact of an antibiotic therapy on the diversity of microbial species. Using next generation sequencing (NGS) analysis of the bacterial 16S ribosomal RNA (rRNA) gene a dominant shift toward Firmicutes and a remodeling in Proteobacteria abundance was observed in the woman's gut microbiota. Considering the patient's age, health status and type of diet, we would have expected to find a GM with a prevalence of Bacteroidetes phylum. This represents the first study investigating the possible TUDCA's effect on human GM.

Comprehensive pan-genome analysis of Lactiplantibacillus plantarum complete genomes.

AIMS: The aim of this work was to refine the taxonomy and the functional characterization of publicly available Lactiplantibacillus plantarum complete genomes through a pan-genome analysis. Particular attention was paid in depicting the probiotic potential of each strain. METHODS AND RESULTS: Complete genome sequence of 127 L. plantarum strains, without detected anomalies, was downloaded from NCBI. Roary analysis of L. plantarum pan-genome identified 1436 core, 414 soft core, 1858 shell and 13,203 cloud genes, highlighting the 'open' nature of L. plantarum pan-genome. Identification and characterization of plasmid content, mobile genetic elements, adaptative immune system and probiotic marker genes (PMGs) revealed unique features across all the L. plantarum strains included in the present study. Considering our updated list of PMGs, we determined that approximatively 70% of the PMGs belongs to the core/soft-core genome. CONCLUSIONS: The comparative genomic analysis conducted in this study provide new insights into the genomic content and variability of L. plantarum. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides a comprehensive pan-genome analysis of L. plantarum, including the largest number (N = 127) of complete L. plantarum genomes retrieved from publicly available repositories. Our effort aimed to determine a solid reference panel for the future characterization of newly sequenced L. plantarum strains useful as probiotic supplements.

Effects of probiotic Lactiplantibacillus plantarum IMC 510 supplementation on metabolic factors in otherwise healthy overweight and obese individuals.

AIMS: Probiotic supplementation approach offers the possibility to shape the gut microbiota (GM), enabling the development of innovative formulations able to improve intestinal well-being and consequently the related body weight modulation and energy metabolism. In the present clinical study, a new potential probiotic supplement based on Lactiplantibacillus plantarum IMC 510 was studied for weight management. METHODS AND RESULTS: Quantitative characterization by qPCR of representative bacterial groups of GM was used to determine the microbiota modulation at different supplementation periods. Furthermore, measurement of the endpoints linked to weight control (body mass index, body weight, waist circumference) was assessed. Specific questionnaires to evaluate the impact on psychological and physiological point of view were performed. Results showed that after 90 days, Lact. plantarum IMC 510 supplementation brought an improvement in endpoints linked to weight control and healthy status, although no significant changes in the microbiota composition were reported for analysed bacterial groups, except for Lactobacillus spp. and Bifidobacterium spp. CONCLUSIONS: We concluded that Lact. plantarum IMC 510 supplementation could be an interesting tool for weight management. More studies are needed to understand the impact on GM, for example, evaluating the production of short-chain fatty acids, since their important role in dietary metabolism. Further research is necessary to better elucidate the relationship between GM and overweight and the mechanism of action by which Lact. plantarum IMC 510 modifies body weight. SIGNIFICANCE AND IMPACT OF THE STUDY: However, these promising outcomes represent a clear advantage of probiotic supplementation and identify a new potential probiotic as a novel and safe therapeutic approach in the obesity prevention and management.

Impact of a probiotic diet on well-being of healthy senior: THE PROBIOSENIOR PROJECT.

AIMS: The aim of this work was to assess the effects of a probiotic diet on well-being of healthy seniors living in boarding and private homes in Marche Region, Italy. In particular, we focused on the modulation of high-sensitivity C-reactive protein (HsCRP), intestinal microbiota and short-chain fatty acids (SCFAs). METHODS AND RESULTS: Ninety-seven healthy seniors took part in a double-blind, placebo-controlled feeding study (59 fed probiotics, 38 fed placebo) for 6 months. Each volunteer ingested daily one food product or a dietary supplement enriched with Synbio® blend (Synbiotec Srl, Camerino, Italy) or the placebo (control group). Blood and faecal samples were collected before and at the end of the intervention period to perform biochemical and microbiological analyses. The serum HsCRP difference value after 6 months of treatment was significantly higher in the probiotic group than placebo (p < 0.05). After the intervention, a significant increase in faecal lactobacilli and a bifidobacteria increase in more participants were observed in the probiotic group. The 16S NGS analysis on the probiotic group showed a decreasing trend of Proteobacteria at the end of the treatment and conversely, an increasing trend of Actinobacteria and Verrucomicrobia phyla, to which the increase of Akkermansiaceae and Bifidobacteriaceae contributes at the family level. Finally, total short-chain fatty acids (SCFAs) and butyric acid were significantly higher in the probiotic group at the end of the treatment respect to the beginning. CONCLUSIONS: Overall, this study emphasizes the beneficial anti-inflammageing effect of a prolonged diet based on functional foods enriched with Synbio® through the modulation of the intestinal microbiota and the consequent increase in the SCFA production. SIGNIFICANCE AND IMPACT OF THE STUDY: Synbio® integration in elderly daily diet may be a preventive strategy to support healthy ageing.

Supplementation with Lactiplantibacillus plantarum IMC 510 Modifies Microbiota Composition and Prevents Body Weight Gain Induced by Cafeteria Diet in Rats.

Changes in functionality and composition of gut microbiota (GM) have been associated and may contribute to the development and maintenance of obesity and related diseases. The aim of our study was to investigate for the first time the impact of Lactiplantibacillus (L.) plantarum IMC 510 in a rat model of diet-induced obesity, specifically in the cafeteria (CAF) diet. This diet provides a strong motivation to voluntary overeat, due to the palatability and variety of selected energy-dense foods. The oral administration for 84 days of this probiotic strain, added to the CAF diet, decreased food intake and body weight gain. Accordingly, it ameliorated body mass index, liver and white adipose tissue weight, hepatic lipid accumulation, adipocyte size, serum parameters, including glycemia and low-density lipoprotein levels, in CAF fed rats, potentially through leptin control. In this scenario, L. plantarum IMC 510 showed also beneficial effects on GM, limiting the microbial imbalance established by long exposure to CAF diet and preserving the proportion of different bacterial taxa. Further research is necessary to better elucidate the relationship between GM and overweight and then the mechanism of action by which L. plantarum IMC 510 modifies weight. However, these promising results prompt a clear advantage of probiotic supplementation and identify a new potential probiotic as a novel and safe therapeutic approach in obesity prevention and management.

Effects of Lactogen 13, a New Probiotic Preparation, on Gut Microbiota and Endocrine Signals Controlling Growth and Appetite of Oreochromis niloticus Juveniles.

In the present study, Nile tilapia Oreochromis niloticus was used as experimental model to study the molecular effects of a new probiotic preparation, Lactogen 13 (Lactobacillus rhamnosus IMC 501® encapsulated with vegetable fat matrices by spray chilling and further indicated as probiotic microgranules), on growth and appetite during larval development. Probiotic microgranules were administered for 30 days to tilapia larvae starting from first feeding. Molecular analysis using high-throughput sequencing revealed that the probiotic could populate the gastrointestinal tract and modulate the microbial communities by significantly increasing the proportion of Lactobacillus as well as reducing the proportion of potential pathogens such as members of the Family Microbacteriaceae, Legionellaceae, and Weeksellaceae. Morphometric analysis evidenced that body weight and total length significantly increased after probiotic treatment. This increase coincided with the modulation of genes belonging to the insulin-like growth factors (igfs) system and genes involved on myogenesis, such as myogenin, and myogenic differentiation (myod). Alongside the improvement of growth, an increase of feed intake was evidenced at 40 days post-fertilization (dpf) in treated larvae. Gene codifying for signals belonging to the most prominent systems involved in appetite regulation, such as neuropeptide y (npy), agouti-related protein (agrp), leptin, and ghrelin were significantly modulated. These results support the hypothesis that gastrointestinal (GI) microbiota changes due to probiotic administration modulate growth and appetite control, activating the endocrine system of tilapia larvae.

Correction to: Effects of Lactogen 13, a New Probiotic Preparation, on Gut Microbiota and Endocrine Signals Controlling Growth and Appetite of Oreochromis niloticus Juveniles.

The original version of this article unfortunately contained mistakes in Authors First name/Family name order. Please refer to this paper as Gioacchini et al. and not as Giorgia et al.

Impact of Probiotic SYNBIO(®) Administered by Vaginal Suppositories in Promoting Vaginal Health of Apparently Healthy Women.

The purpose of this study was to investigate whether vaginal administration of probiotic Lactobacillus results in their colonization and persistence in the vagina and whether it promotes normalization and maintenance of pH and Nugent score. A single-arm, open-label controlled towards the baseline (pre-post) study including 35 apparently healthy women was conducted. Each woman was examined three times during the study. Women were instructed to receive daily for 7 days, the probiotic suppositories SYNBIO(®) gin (Lactobacillus rhamnosus IMC 501(®) and Lactobacillus paracasei IMC 502(®)). Vaginal swabs were collected during visit 1, 2, and 3 to determine the total lactobacilli count, the presence of the two administered bacteria, the measure of the pH, and the estimation of Nugent score. Evaluation of treatment tolerability was based on analysis of the type and occurrence of adverse events. The probiotic vaginal suppository was well tolerated and no side effects were reported. Intermediate Nugent score was registered in 40 % of women at visit 1 and these intermediate scores reverted to normal at day 7 (end of treatment) in 20 % of subjects. Administration of SYNBIO(®) gin contributed to a significant increase in the lactobacilli level at visit 2. Molecular typing revealed the presence of the two strains originating from SYNBIO(®) gin in 100 % of women at visit 2 and 34 % at visit 3. No significant changes were registered for pH between visits. The SYNBIO(®) gin product is safe for daily use in healthy women and it could be useful to restore and maintain a normal vaginal microbiota.

Probiotic-enriched foods and dietary supplement containing SYNBIO positively affects bowel habits in healthy adults: an assessment using standard statistical analysis and Support Vector Machines.

A randomised, double-blind, placebo-controlled, parallel group study assessed in healthy adults how daily consumption of the probiotic combination SYNBIO®, administered in probiotic-enriched foods or in a dietary supplement, affected bowel habits. Primary and secondary outcomes gave the overall assessment of bowel well-being, while a Psychological General Well-Being Index compiled by participants estimated the health-related quality of life as well as the gastrointestinal tolerance determined with the Gastrointestinal Symptom Rating Scale. Support Vector Machine models for classification problems were used to validate the total outcomes on bowel well-being. SYNBIO® consumption improved bowel habits of volunteers consuming the probiotic foods or capsules, while the same effects were not registered in the control groups. The recovery of probiotic bacteria from the faeces of a cohort of 100 subjects for each supplemented group showed the persistence of strains in the gastrointestinal tract.

In Vitro Assessment of Postbiotic and Probiotic Commercial Dietary Supplements Recommended for Counteracting Intestinal Dysbiosis in Dogs.

Many environmental aspects influence the preservation of a beneficial microbiome in dogs, and gut dysbiosis occurs when imbalances in the intestinal ecosystem cause functional changes in the microbial populations. The authors evaluated the effects of two specific commercial dietary supplements: a combination of a postbiotic and prebiotics (Microbiotal cane®) and a probiotic product (NBF 1®) recommended for counteracting intestinal dysbiosis in dogs, on the gut canine microbiota composition and its metabolic activities (production of short-chain fatty acids). The investigation was performed using an in vitro fermentation system inoculated with dog fecal samples. Microbiotal cane® promoted a more immediate increase in Lactobacillus spp. after the first 6 h of fermentation, whereas NBF 1® promoted the increase at the end of the process only. The two supplements supported an increase in the Bifidobacterium spp. counts only after 24 h. The in vitro abilities of Microbiotal cane® and NBF 1® to increase selectively beneficial bacterial groups producing acetic, propionic, and butyric acids suggest a possible positive effect on the canine gut microbiota, even if further in vivo studies are needed to confirm the beneficial effects on the intestinal health.

SYNBIO® Probiotic and Antioxidant Dietary Supplementation: Clinical Trial Evaluation of Potential Effects on Airline Flight Crew Members' Well-Being.

The irregular lifestyle of airline crew members, wide/adverse job-related exposures, and the impact of temporary hypoxia on gut microbiota well-being have increased concern about the daily recommended dose of certain nutrients among flight crew. The aim of this study was to determine if daily consumption of a SYNBIO® probiotics-elderberry extract supplement (ACTIVE) may contribute to the well-being of flight attendants. Forty healthy crew members enrolled in a double-blind, randomized, placebo-controlled study consumed one ACTIVE capsule/day or placebo for 30 days. Bowel well-being, health-related quality of life, and gastrointestinal tolerance were assessed by validated questionnaires. Saliva and fecal samples were analyzed to determine secretory immunoglobulin-A (sIgA) levels and to characterize gut microbiota composition, respectively. ACTIVE subjects presented a physiological improvement and a statistically significant higher Psychological General Well-Being Index (PGWBI) global score compared to PLACEBO subjects. The ACTIVE subjects showed significantly increased levels of lactobacilli and bifidobacteria compared to the PLACEBO group, while a significant increase in lactobacilli and a significant reduction in Enterobacteriaceae were registered when compared with the beginning of supplementation, confirming the persistence of probiotics in the gastrointestinal tract and the direct antagonism and competitive exclusion effects. Additionally, sIgA levels were significantly higher in the ACTIVE group compared to the baseline and to the PLACEBO group at the end of supplementation. The ACTIVE supplementation might be beneficial to airline crew members, improving their physiological state, their immune defenses, and the strength and efficiency of their gastrointestinal tract when responding to stressful conditions.

Six-Month Synbio® Administration Affects Nutritional and Inflammatory Parameters of Older Adults Included in the PROBIOSENIOR Project.

The physiological changes associated with ageing contribute to the incidence of diseases, morbidity, and mortality. For modern society, it is essential to find solutions to improve elderly people's health and quality of life. Among promising strategies, the PROBIOSENIOR project proposed a daily six-month supplementation with new probiotic functional foods and nutraceuticals. The aim of this work was to evaluate the modulating effects of the probiotic diet on inflammatory markers and nutritional status. Ninety-seven elderly volunteers were randomly assigned to either a placebo-diet group or a probiotic-diet group (SYNBIO®). Faeces, urine, and blood samples were collected before and after the supplementation to determine serum cytokines, biogenic amines, and inflammation markers. Comparing the results obtained before and after the intervention, probiotic supplementations significantly decreased the TNF-α circulating levels and significantly increased those of IGF-1. Biogenic-amine levels showed high variability, with significant variation only for histamine that decreased after the probiotic supplementation. The supplementation influenced the serum concentration of some crucial cytokines (IL-6, IL-8, and MIP-1α) that significantly decreased in the probiotic group. In addition, the Mini Nutritional Assessment questionnaire revealed that the probiotic-supplemented group had a significant improvement in nutritional status. In conclusion, the PROBIOSENIOR project demonstrated how SYNBIO® supplementation may positively influence some nutritional and inflammatory parameters in the elderly.

Antimicrobial efficacy of Achillea ligustica All. (Asteraceae) essential oils against reference and isolated oral microorganisms.

The aim of this study was to verify the effectiveness of Achillea ligustica essential oils against several oral microorganisms in comparison with a commercial essential oil-containing mouthrinse (Listerine(®)) and clove oil (containing 89% eugenol). The inhibition efficacy of A. ligustica essential oils alone and in combination with Listerine(®) was evaluated by the micro-dilution method. The most susceptible microorganisms were Bacillus cereus, Streptococcus pyogenes, and Candida albicans. The efficacy was similar to that of the clove oil. The antiseptic mouthwash Listerine(®) did not exert a strong inhibition on microbial strains tested, whereas its effectiveness increased significantly when essential oil was added. The study provides additional evidence for the in vitro inhibitory activity of A. ligustica essential oils on several pathogens, suggesting their usefulness in mouthrinse formulations as an adjunct to mechanical oral hygiene regimens. Essential oil-containing mouthrinses can be beneficial, safe components of daily oral health routines, representing an efficient and without side effect alternative to prevent and control oral infections.

Influence of modified governing liquid on shelf-life parameters of high-moisture mozzarella cheese.

The roles of mesophilic lactobacilli in cheese manufacturing and ripening have been widely studied, but their impacts, especially as additives in preserving liquids, on the high-moisture mozzarella cheese quality parameters remained underexplored. The current study studied and compared the effects of four preserving liquid formulations - brine solution (sodium chloride, as control), a salt mixture solution, and two lactobacilli solutions (Lactiplantibacillus plantarum IMC 509 in brine solution) or SYNBIO® (a 1:1 ratio of Lacticaseibacillus rhamnosus IMC 501 and Lacticaseibacillus paracasei IMC 502) - on the microbial, chemical (volatile fatty acids), physicochemical (moisture, weight, pH, colour), texture (adhesiveness, hardness, cohesiveness, gumminess, springiness, chewiness) and sensorial qualities of mozzarella stored for 30 days at 4 °C. The quality of the governing solution, including microbial content, lactobacilli viability, pH, turbidity, and smell, was also monitored. For the first 10 days of storage, all samples demonstrated similar physicochemical variations: the firmness and chewiness of the mozzarella lessened, the pH values of the liquid decreased, but cheese microbial growth increased, as did the levels of free fatty acids, the mozzarella pH values and skin whiteness. At day 20, mozzarella stored in mesophilic lactobacilli liquid, especially SYNBIO®, had a spongy structure, tasted more bitter and sour, and had lower pH values than mozzarella in the simple brine solution. Moreover, the cheese sample and liquid of SYNBIO® also presented higher coliform and Pseudomonas spp. counts than that detected in the control, the SYNBIO® liquid, especially in the later period of storage, exhibited a yogurt smell and increased turbidity. By contrast, Mozzarella packed into salt mixture liquid exhibited somewhat more adhesiveness and gumminess, a saltier taste and slightly higher microbial counts than the control sample. The mozzarella samples differed slightly in colour and texture as well. The overall quality changes suggested that the use of the studied mesophilic lactobacilli as governing liquid additives may not be appropriate for high-moisture mozzarella cheese preservation.

A shelf-life study for the evaluation of a new biopackaging to preserve the quality of organic chicken meat.

Widespread use of traditional packaging constitutes a serious ecological problem leading to a shift to biodegradable and compostable materials. The aim of this work is to study the ability of a new biopackaging (BP), based on biodegradable and compostable material, to preserve the quality of organic chicken meat for 14 days in comparison with a polyethylene terephthalate (PET) material. Results showed that the indices of Biogenic Amines (BAs) and the 18 monitored Volatile Organic Compounds (VOCs) have a similar trend in both packaged meats. For example, the total BAs concentration in meat increased from 390 to 961 mg Kg-1 in BP and from 393 to 800 mg Kg-1 in PET, as well as the microbiological counts. The new biopackaging (BP) showed similar properties of non-biodegradable material (PET) to preserve the shelf life of organic chicken meat and it could be used instead of plastic materials to promote a circular economy.

Effect of a probiotic intake on oxidant and antioxidant parameters in plasma of athletes during intense exercise training.

The aim of this study was to evaluate the effect of Lactobacillus rhamnosus IMC 501 and Lactobacillus paracasei IMC 502 on oxidative stress in athletes during a four-week period of intense physical activity. Two groups of twelve subjects each were selected for this analysis. The first group consumed a daily dose of a mixture of the two probiotic strains (1:1 L. rhamnosus IMC 501 and L. paracasei IMC 502; ~10(9) cells/day) for 4 weeks. The second group (control) did not consume any supplements during the 4 weeks. Blood samples collected immediately before and after the supplementation were analyzed, and plasma levels of reactive oxygen metabolites and biological antioxidant potential were determined. Faeces were also collected and analyzed before and at the end of the probiotic supplementation. Antioxidative activity and oxidative stress resistance of the two strains were determined in vitro. Results demonstrated that intense physical activity induced oxidative stress and that probiotic supplementation increased plasma antioxidant levels, thus neutralizing reactive oxygen species. The two strains, L. rhamnosus IMC 501(®) and L. paracasei IMC 502(®), exert strong antioxidant activity. Athletes and all those exposed to oxidative stress may benefit from the ability of these probiotics to increase antioxidant levels and neutralize the effects of reactive oxygen species.

Modifying the Silver-Titania Nanocomposites with Carbonaceous Materials to Remove the Pollutants from Domestic Waste Water.

The aim of this study was to prepare and characterize nanostructured composites based of TiO2, carbonaceus materials (GN or GO) and Ag and the test their capacity to remove the pollutants from domestic wastewater. The composites were characterized by IR and UV-Vis spectroscopy, X-ray diffraction, electron microscopy and nitrogen adsorption-desorption measurements. The photocatalytic activity was measured from the experiment of salicylic acid (SA) degradation. The capacity to remove the pollutants from domestic wastewater was performed by considering the absorbance of residual solution at 200 nm. The non-calcined composites have high specific surface area (˜300 m²/g), but nitrogen adsorption-desorption isotherms showed a porous structure with closed pores. The porosity of the thermal treated composites is about 10 times less, but the pores are open. The salicylic acid was 94% degraded over all composites, showing their efficient photoactivity. A percent of 70% of pollutants were removed over the calcined composites with GN and ˜67% on those with GO. It was no statistically significant difference between the photocatalytical efficiency of GN- and GO-based composites. Even if the calcined composites have the specific surface area about 10 times lower, their lower gap energy, higher degree of crystallinity and photocatalytic activity make them efficient candidates for removal of pollutants from domestic waste water. The photodegradation mechanism occurred mostly by π-π interactions between GN/GO and pollutant molecules.

Chemical and organoleptic changes of curd cheese stored in new and reused active packaging systems made of Ag-graphene-TiO2-PLA.

The study reports obtaining, characterization and evaluation of the preservation efficacy of nano-Ag-graphene-TiO2-polylactic acid (PLA) film during the storage of the curd cheese. The reusability of the active package was also evaluated. The mechanical resistance of the film was improved by 30% following the composite addition. Water vapour permeability decreased by 11-27% when the composite was incorporated into PLA in a mass percentage ranged between 0.5 and 3%. The highest fat permeability was obtained for PLA3% at 4 °C. The oxygen permeability of PLA3% is 24.6% lower than that of neat-PLA. PLA0.5% and PLA3% presented the highest antibacterial activity. PLA3% achieved the lowest solubility in food simulants at 4 °C compared to PLA and reference. The best active packages for curd storage were PLA0.5% and PLA3%. They showed also the lowest depreciation from the first to the second use. Their active role is kept 100% and 85% respectively, during the second use.

Effect of dietary probiotics on clownfish: a molecular approach to define how lactic acid bacteria modulate development in a marine fish.

We set out to determine whether probiotic addition would improve larval development in the false percula clownfish Amphiprion ocellaris and to determine what molecular responses could be observed in the larvae following probiotic exposure. We supplied the probiotic bacterial strain Lactobacillus rhamnosus IMC 501 to clownfish larvae from the first day posthatch simultaneously by live prey and with addition to rearing water (group 2) and exclusively by live prey (group 3). We observed twofold higher body weight in both clownfish larvae and juveniles when probiotics were supplied via live prey and added to the rearing water. In addition, development was accelerated with metamorphosis occurring 3 days earlier in fingerlings treated with probiotic. Alteration in molecular biomarkers supported the faster growth observation. There was significantly increased gene expression of factors involved in growth and development (insulin-like growth factors I and II, myostatin, peroxisome proliferator-activated receptors alpha and beta, vitamin D receptor alpha, and retinoic acid receptor gamma) when probiotics were delivered via live prey and added to the rearing water. Moreover, probiotic treatment lessened the severity of the general stress response as exhibited by lower levels of glucocorticoid receptor and 70-kDa heat shock protein gene expression. Furthermore, an improvement of skeletal head development was observed, with a 10-20% reduction in deformities for juveniles treated with probiotic. All data suggest a potent effect on development resulting from the administration of lactic acid bacteria to larval clownfish, and this study provides a preliminary molecular entry path into the investigation of mechanisms responsible for probiotic enhancement in fish development.