Gut dysbiosis, inflammation and type 2 diabetes in mice using synthetic gut microbiota from diabetic humans

The study was aimed to assess impact of high fat diet (HFD) and synthetic human gut microbiota (GM) combined with HFD and chow diet (CD) in inducing type-2 diabetes (T2D) using mice model. To our knowledge, this is the first study using selected human GM transplantation via culture based method coupled dietary modulation in mice for in vivo establishment of inflammation leading to T2D and gut dysbiosis. Twenty bacteria (T2D1-T2D20) from stool samples of confirmed T2D subjects were found to be morphologically different and subjected to purification on different media both aerobically and anerobically, which revealed seven bacteria more common among 20 isolates on the basis of biochemical characterization. On the basis of 16S rRNA gene sequencing, these seven isolates were identified as Bacteroides stercoris (MT152636), Lactobacillus acidophilus (MT152637), Lactobacillus salivarius (MT152638), Ruminococcus bromii (MT152639), Klebsiella aerogenes (MT152640), Bacteroides fragilis (MT152909), Clostridium botulinum (MT152910). The seven isolates were subsequently used as synthetic gut microbiome (GM) for their role in inducing T2D in mice. Inbred strains of albino mice were divided into four groups and were fed with CD, HFD, GM+HFD and GM+CD. Mice receiving HFD and GM+modified diet (CD/HFD) showed highly significant (P<0.05) increase in weight and blood glucose concentration as well as elevated level of inflammatory cytokines (TNF-α, IL-6, and MCP-1) compared to mice receiving CD only. The 16S rRNA gene sequencing of 11 fecal bacteria obtained from three randomly selected animals from each group revealed gut dysbiosis in animals receiving GM. Bacterial strains including Bacteroides gallinarum (MT152630), Ruminococcus bromii (MT152631), Lactobacillus acidophilus (MT152632), Parabacteroides gordonii (MT152633), Prevotella copri (MT152634) and Lactobacillus gasseri (MT152635) were isolated from mice treated with GM+modified diet (HFD/CD) compared to strains Akkermansia muciniphila (MT152625), Bacteriodes sp. (MT152626), Bacteroides faecis (MT152627), Bacteroides vulgatus (MT152628), Lactobacillus plantarum (MT152629) which were isolated from mice receiving CD/HFD. In conclusion, these findings suggest that constitution of GM and diet plays significant role in inflammation leading to onset or/and possibly progression of T2D. .

Bacterial viability, antioxidant stability, antimutagenicity and sensory properties of onion types fermentation by using probiotic starter during storage

Purpose The purpose of this study was to assess the bacterial viability, antioxidative activity, antimutagenicity and sensory evaluation of fermented onion types by using probiotic starters after fermentation at 37 °C for 24 hours and storage in the refrigerator for 28 days. Design/methodology/approach For onion fermentation, Lactobacillus acidophilus (LA-5), Bifidobacterium bifidum (BB-12), and Streptococcus thermophilus (ST) were utilised. This research was conducted on three types of onion: white onion, red onion and scallion. With a 5% brine solution, the onions were sliced into 3-5 cm long and 1-2 cm wide slices. The process of fermentation was achieved by adding 2% (108 CFU/ gm) of fresh probiotic starter and incubating it for 24 hours at 37 °C. The fermented onion samples were kept in the refrigerator for 28 days. After fermentation and storage, the pH and total acidity were estimated, the vitality of probiotic bacteria was evaluated in samples of the onion species. The Gas chromatography-mass spectrometry (GC-MS) technique was used to identify the bioactive components in fermented onion types. The antioxidant activity of fermented onions was measured using the 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical scavenging activity assay and the hydroxyl radical scavenging activity test. The Ames test was used to detect the antimutagenicity of fermented onion samples. Findings After fermentation, the fermented scallion (p = 0.036) has the highest vitality of all the starter bacteria species. The fermentation of onion types produced a pH of between 4.1–4.7 and 0.19–0.23% total acidity, which is in the range of reduced 3.1–3.5 pH values and 0.42–0.63% total acidity after 28 days. The viability of Lactobacillus acidophilus and Bifidobacterium bifidum in fermented scallions was Log. 7.79 and 7.57 CFU/gm. The GC-MS technique found 14 bioactive compounds in fermented white onions and 13 compounds in fermented white onions, with 15 compounds in scallion fermentation. The majority of these bioactive compounds are strong antioxidants. The antioxidant properties of fermented scallion significantly increased after 28 days of storage time, showing an inhibitory effect on the DPPH assay (p = 0.02) and the scavenging activity of the hydroxyl radical assay (p = 0.01). Sensory evaluation tests revealed that the fermented scallion was a suitable product in terms of appearance, aroma and overall acceptability. Originality/value Commercially accessible probiotic foods account for a sizable portion of the consumer market. Furthermore, as consumer interest in healthy eating grows, so does demand for plant-based goods. All onion types fermented with probiotic bacteria have many chemical compounds that have both antioxidant and carcinogenic activity. The fermented scallion onion sample was significantly superior to the rest of the other types of onions.

Antibacterial efficacy of spice extracts against Streptococcus mutans and Lactobacillus acidophilus – An in-vitro study

Objectives: To evaluate the antibacterial activity of black pepper, Indian bay leaf, cinnamon, and cumin against Streptococcus mutans and Lactobacillus acidophilus in-vitro and to determine their minimum inhibitory concentration (MIC). Materials and Methods: The spices (cinnamon, cumin, Indian bay leaf, and black pepper) were obtained from local market, were dried and powdered. Solvent extracts were prepared with methanol by maceration, followed by filtration and evaporation. The antimicrobial activity was assessed using cup plate diffusion method, followed by determination of MIC of the extracts. Statistical analysis was performed using one-way analysis of variance and Tukey’s post hoc test was used for pairwise comparison. P < 0.05 was considered statistically significant. Results: All the four extracts showed significant antimicrobial activity. Cinnamon demonstrated maximum activity against S. mutans (zone of inhibition of 18.1 mm ± 0.30) and L. acidophilus (zone of inhibition of 17.9 mm ± 0.44) with the least MIC against the organisms (<0.05 mg/ml). Conclusion: All the spice extracts tested demonstrated significant antibacterial activity against S. mutans and L. acidophilus. On comparison of the antibacterial activities of all the four extracts, cinnamon extract emerged as the potent agent.

The Use of Olkuska Sheep Milk for the Production of Symbiotic Dairy Ice Cream

The aim of this study was to determine the possibility of using Olkuska sheep milk for the production of ice cream with probiotics and prebiotics. The study examined the effect of the storage and type of bacteria used for the fermentation of ice cream mixes and partial replacement of inulin with apple fiber on the physicochemical properties, viability of probiotic cultures and organoleptic properties of sheep’s milk ice cream stored at −22 °C for 21 days. The addition of apple fiber reduced the pH value of ice cream mixes before fermentation. In ice cream mixes and ice cream with apple fiber, the lactic acid content was higher by 0.1–0.2 g L−1 than in their equivalents with inulin only. These differences persisted during the storage of the ice cream. After fermentation of the ice mixes, the bacterial cell count ranged from 10.62 log cfu g−1 to 12.25 log cfu g−1. The freezing process reduced the population of probiotic bacteria cells in ice cream with inulin from 0.8 log cfu g−1 in ice cream with Lactobacillus acidophilus, 1.0 log cfu g−1 in ice cream with Lacticaseibacillus paracasei and 1.1 log cfu g−1 in ice cream with Lacticaseibacillus casei. Freezing the varieties with apple fiber also resulted in a reduction of viable bacterial cells from 0.8 log cfu g−1 in ice cream with L. paracasei and Lb. acidophilus to 1 log cfu g−1 in ice cream with L. casei, compared to the results after fermentation. The highest percentage overrun was determined in ice cream with L. paracasei and Lb. acidophilus. Ice cream with L. casei was characterized by significantly lower overrun on the 7th and 21st days of storage. Although L. paracasei ice cream had the highest overrun, it did not cause a significant reduction in the probiotic population during storage. After seven days of storage, the first drop differed significantly depending on the type of bacteria used for fermentation of the mixture and the addition of apple fiber. L. casei ice cream had a longer first drop time than L. paracasei and Lb. acidophilus ice cream. Partial replacement of inulin with apple fiber resulted in a significant darkening of the color of ice cream mixes. Depending on the type of bacteria used for fermentation, the addition of apple fiber decreased the value of the L* parameter. Ice cream mixes and ice cream with inulin and apple fiber were characterized by a high proportion of yellow. Partial replacement of inulin with apple fiber reduced the hardness of ice cream compared to inulin-only ice cream. Moreover, the panelists found that ice cream with inulin was characterized by a sweeter taste than ice cream with apple fiber. Moreover, the addition of apple fiber favorably increased the flavor and aroma perception of the mango-passion fruit. Therefore, the milk of Olkuska sheep could be successfully used for the production of symbiotic dairy ice cream.