Effect of Dietary Probiotic Lactobacillus helveticus on Growth Performance, Antioxidant Levels, and Absorption of Essential Trace Elements in Goldfish (Carassius auratus).

In this study, we have investigated the effect of probiotic Lactobacillus helveticus CD6 supplemented into simple laboratory fish feed (LFF) and complex, multi-ingredient market fish feed (MFF) on growth performance, antioxidant levels, and essential trace element absorption in goldfish (Carassius auratus). Twenty-four healthy goldfish (average weight 3-4 g) were acclimatized and divided into four experimental diets supplemented with 3 × 107 CFU/g of probiotic (LFF + Pro, MFF + Pro) and without probiotic (LFF, MFF) for 14 weeks. Fish fed with probiotic were healthy, active, and intense orange-gold as compared to control (without probiotic). At 14 weeks, fish fed with MFF + Pro/LFF + Pro showed 91/47% weight gain as compared to 34/- 12% weight observed with MFF/LFF. The average weight gain differences recorded between probiotic and control diets were not significant. No mortality to report when fish fed with probiotic. In contrast, fish fed without probiotic showed mortalities (LFF, two fish; MFF, one fish) during the trial. DPPH activity revealed high levels of antioxidants into the intestine of probiotic-fed fish. Trace element analysis showed that probiotic colonization enhanced diet-dependent absorption of Fe and Zn. The in vitro antimicrobial activity was exhibited by probiotic L. helveticus CD6 against infected fish isolate Aeromonas spp. JA showed an ability to protect fish from infections. Moreover, complex, multi-ingredient feed had a highest impact on viability of probiotic during storage. In conclusion, L. helveticus CD6 did not significantly enhance growth performance; however, it improved health and reduced mortalities in goldfish (C. auratus) regardless of the composition of the diet.

In vitro and in vivo antagonistic activity of new probiotic culture against Clostridium difficile and Clostridium perfringens.

BACKGROUND: Genus Clostridium accompanies more than 200 known species and at least 30 among them are associated with human and animal diseases. At the moment, the treatment of clostridial infections is based on use of antibiotics. However, due to the European ban on the use of antibiotics in livestock production, novel therapeutic strategies for treatment of these hardly curable infections have been evaluated. Hence, in this study the antimicrobial effect of newly designed probiotic culture consisted of natural isolates Lactobacillus helveticus BGRA43, Lactobacillus fermentum BGHI14 and Streptococcus thermophilus BGVLJ1-44 against Clostridium difficile and Clostridium perfringens was analyzed. RESULTS: The probiotic culture showed strong in vitro antimicrobial effect on C. difficile (human clinical isolate). In addition, individual strains and the probiotic combination exhibited immunomodulatory activity. The probiotic combination significantly increased the proliferation of GALT lymphocytes. At the other hand, none of the bacterial treatments (individual strains and the combination) induced the production of proinflammatory cytokines IL-6 and IL-1ß by intestinal epithelial cells, Caco-2. Interestingly, Caco-2 cells exposed to the probiotic combination produced significantly elevated amount of TGFß pointing to potential protecting effect of the probiotic. In addition, the results of field trial on spontaneously infected goats revealed reduction of C. perfringens in goats (below the detection threshold) after the probiotic treatment. CONCLUSIONS: The results of this study indicated that the novel probiotic deserves to be further investigated as a promising antimicrobial agent against C. difficile and C. perfringens.

Randomized clinical trial: efficacy of a food supplement, TRANSITECH, on healthy individuals with mild intermittent constipation.

BACKGROUND: Constipation is a common disorder in the general population and can be observed in healthy individuals. A natural product leading to an increase in bowel movements and decrease in colonic transit time (CTT), without bloating, could be useful for the patient's care. OBJECTIVES: To investigate the effects of TRANSITECH, a food supplement composed of plants and lactic ferments, on bowel movements, CTT and bloating. METHODS: A total of 100 healthy participants, presenting two to five stools per week, were selected and followed over a 6-day baseline period. They were randomly assigned to receive daily two tablets of TRANSITECH or placebo during 10 days. They were then followed up over 28 days after intervention. Participants daily recorded in a home questionnaire the characteristics of stools (frequency and consistency), and the importance of bloating during the preintervention period (from D-6 to D0), the intervention period (from D0 to D10) and the postintervention period (from D10 to D38). Their CTTs were also evaluated by following the propagation of radiopaque markers at D0 and D10. RESULTS: At D10, the food supplement group showed, compared with the placebo group, higher daily stool emission (0.95±0.50, 0.70±0.20, P<0.001), softer stool consistency (2.5±0.6 vs. 3.0±0.8, P<0.001) and lower CTT (33.8±28.2 vs. 56.4±36.2 h, P=0.01). The active group also showed a sustained increase in daily stool emissions observed at D38 compared with D0 (P=0.03). CONCLUSION: TRANSITECH is an efficient natural solution for the treatment of constipation. It increases the number of bowel movements, decreases the oroanal and segmental CTT, is well tolerated, and presents sustained effects after treatment completion.

Effects of Lactobacillus helveticus on murine behavior are dependent on diet and genotype and correlate with alterations in the gut microbiome.

Modulation of the gut microbiota with diet and probiotic bacteria can restore intestinal homeostasis in inflammatory conditions and alter behavior via the gut-brain axis. The purpose of this study was to determine whether the modulatory effects of probiotics differ depending on diet and mouse genotype. At weaning, wild type (WT) and IL-10 deficient (IL-10(-/-)) 129/SvEv mice were placed on a standard mouse chow or a Western-style diet (fat 33%, refined carbohydrate 49%)±Lactobacillus helveticus ROO52 (10(9)cfu/d) for 21 days. Animal weight and food eaten were monitored weekly. Intestinal immune function was analysed for cytokine expression using the Meso Scale Discovery platform. Spatial memory and anxiety-like behavior was assessed in a Barnes maze. Terminal restriction fragment length polymorphism (TRFLP) was used to analyze the fecal microbiota. Both WT and IL-10(-/-) mice on a Western diet had increased weight gain along with changes in gut microbiota and cytokine expression and altered anxiety-like behavior. The ability of L. helveticus to modulate these factors was genotype- and diet-dependent. Anxiety-like behavior and memory were negatively affected by Western-style diet depending on inflammatory state, but this change was prevented with L. helveticus administration. However, probiotics alone decreased anxiety-like behavior in WT mice on a chow diet. Mice on the Western diet had decreased inflammation and fecal corticosterone, but these markers did not correlate with changes in behavior. Analysis of bacterial phyla from WT and IL-10(-/-)mice showed discrete clustering of the groups to be associated with both diet and probiotic supplementation, with the diet-induced shift normalized to some degree by L. helveticus. These findings suggest that the type of diet consumed by the host and the presence or absence of active inflammation may significantly alter the ability of probiotics to modulate host physiological function.

Use of ß-galactosidase liposome model as a novel method to screen freeze-drying cryoprotectants.

This study developed a novel method of screening cryoprotectants used to improve the survivability of lyophilized Lactobacillus helveticus. To develop a liposome encapsulated ß-galactosidase (ß-gal) as a cell membrane model, the ß-gal liposome was characterized in terms of mean size, poly dispersity index, zeta potential, along with transmission electron microscopy. 800 W of ultrasonic power and 10 min of sonication time were the optimal experimental conditions to obtain the desirable ß-gal liposome. Subsequently, different cryoprotectants were mixed with the ß-gal liposome during freeze-drying. After freeze-drying, liposomes were hydrolized, and the protective effect of cryoprotectants was assessed as the release rate of encapsulated ß-gal. The lowest release rate of ß-gal was obtained using 10 mg/100 ml trehalose and 0.2 mg/100 ml hyaluronic acid.