Bacillus toyonensis SAU-19 Ameliorates Hepatic Insulin Resistance in High-Fat Diet/Streptozocin-Induced Diabetic Mice

Insulin resistance (IR) is a hallmark of type 2 diabetes mellitus (T2DM). This study was performed to investigate the antidiabetic effect of Bacillus toyonensis SAU-19 and its possible mechanisms of action in mice with type 2 diabetes mellitus (T2DM). Thirty SPFKM mice were randomly assigned to three groups: control, diabetic model, and diabetes + Bacillus toyonensis SAU-19 group. After 35 days, blood was collected for biochemical analysis and liver tissue samples for histopathological analysis using H&E staining, qPCR, and ELISA. The results showed that the administration of B. toyonensis SAU-19 significantly improved the blood glucose, hepatic insulin resistance, and morphological changes of the liver characterized by significant improvement of dyslipidemia, glycogen synthesis, and antioxidant status (p < 0.05), indicating the strains’ ameliorating effects on hepatic insulin resistance in T2DM. In conclusion, the probiotic strain (B. toyonensis SAU-19) inhibits T2DM by reducing insulin resistance, improving antioxidant status, and downregulating genes related to glucose synthesis; hence, it may be used in treating diabetes and other metabolic disorders. This study provides the basis for further studies into the molecular mechanisms of B. toyonensis SAU-19 in treating T2DM.

Growth of Bacillus toyonensis in Tofu Wastewater

Bacillus toyonensis has been isolated in Dumai mangrove ecosystem of Riau Province. One of factors affecting the growth of the bacteria is growth substrate. Tofu wastewater is rich in nutrition which can be used as substrate for bacterial growth. This research aimed to observe the growth of B. toyonensis in different concentration of tofu wastewater. The bacteria was grown in tofu wastewater at concentrations 8%, 10% and 12% was supplemented with 0.1 g K2HPO4, 0.15 g KH2PO4, 0.15 g NaCl and 0.5 g vitamin B12 in 100 mL distilled water. The bacterial growth was observed by using spectrophotometer at λ 610 nm and by analysis the total plate counts on plate count agar (PCA) at 0, 24, 48, 72 and 96 hour cultivation. Spectrophotometric observation showed that the highest bacterial growth of all tofu wastewater treatments indicated by the addition of 12% tofu wastewater, although the absorbance value was lower than culture in tryptic soy broth (TSB) as control. Exponential growth occurred between 0-24 hour incubation, and the highest growth indicated in substrate contained 12% tofu wastewater. Similarly, total plate count (TPC) analysis indicated that the highest bacterial growth of all treatment occurred at 24 hours incubation, and the highest count was also indicated by treatment of 12% tofu wastewater (2.42±0.06×108 CFU/mL). In conclusion, tofu wastewater can be an alternative substrate for the bacterial growth.

Development of a probiotic for animals and aquaculture based on Bacillus toyonensis B-13249 and Bacillus pumilus B-13250 strains

This article aims to develop a probiotic for animals and aquaculture based on the Bacillus toyonensis B-13249 and Bacillus pumilus B-13250 strains. The selection of a nutrient medium was conducted for cultivating the inoculum of these microorganisms. Several bacteria fermentations of the Bacillus genus were performed in biological reactors with a capacity of 15 and 250 l. A technology for obtaining a finished probiotic for animals and aquaculture was developed. The results indicate that L-broth is the most optimal nutrient medium for cultivating the studied strains. The cultivation of B. toyonensis B-13249 and B. pumilus B-13250 strains in fermenters revealed that sporulation begins after 4–8 hours of fermentation. In contrast to the vegetative medium, the fermentative medium helped the bacilli develop a higher optical density (the maximum value in the B. pumilus strain – 2.400±0.149), pH value (maximum value in the B. toyonensis strain – 8.483±0.609) and titer (at least 1010 CFU/g). After 20–24 hours of incubation, both strains of bacilli in the fermenter, almost completely pass into endospores, which serve as a signal for the start of biomass centrifugation. This was indicated by the following: from a 15 l fermenter – 83.3±6.1 g of concentrate, from a 250 l fermenter – 499.8±51.4 g. The number of bacilli in a concentrated state was at least 1·1011 CFU/g for both strains. Obtaining a finished preparation required mixing bacterial concentrates with maltodextrin to a titer of at least 1·1010 CFU/g. The number of bacteria in the preparation checked every month during the year, recorded no value less than 1·1010 CFU/g. Thus, L-broth is most favorable for growing the mother culture of the B. toyonensis B-13249 and B. pumilus B-13250 strains, and fermentative nutrient medium – for the cultivation in fermenters. The expiry date of the bacilli-based biological preparation is at least 12 months, during which the drug’s polycomponence, color and consistency are preserved, in addition to the bacteria titer (at least 1·1010 CFU/g) and their viability.

Bioprospecção e caracterização de bactérias produtoras de celulase a partir do solo do Cerrado brasileiro

Este trabalho descreve a bioprospecção e a caracterização de bactérias produtoras de celulase isoladas de solo obtido em uma região preservada do bioma Cerrado, em Uberaba, Brasil. A atividade celulolítica foi confirmada em 14 das 84 linhagens bacterianas isoladas. De acordo com a quantificação enzimática, cinco isolados foram selecionados como os melhores produtores de celulase e com base no sequenciamento parcial do gene 16S rRNA foram identificadas como Bacillus siamensis (isolado AB-9; 5.000U/mL), Bacillus toyonensis (isolado AB-1; 4.630U/mL), Bacillus methylotrophicus (isolados MB-3; 4.236U/mL e MP-7; 4.282U/mL) e Bacillus drentensis (isolado ME-2; 4.444U/mL). O extrato enzimático obtido de B. siamensis AB-9 foi o mais estável em diferentes valores de pH (2-8), mantendo sua atividade celulolítica, enquanto B. toyonensis AB-1 e B. methylotrophicus MP-7 produziram celulases com atividade máxima em pH 7 e 8. As celulases produzidas por B. siamensis AB-9 e B. toyonensis AB-1 apresentaram alta atividade enzimática em todas as temperaturas analisadas (10-80°C), enquanto as celulases de B. methylotrophicus MB-3 apresentaram atividade máxima na faixa de 20-70°C. Até o momento, esta é a primeira vez que bactérias produtoras de celulase isoladas do bioma Cerrado brasileiro na região do Triângulo Mineiro com potencial aplicação biotecnológica são descritas.

Optimization and Degradation Studies on Hexahydro-1,3,5-Trinitro-1,3,5-Triazine (RDX) with Selected Indigenous Microbes under Aerobic Conditions

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) a nitramine explosive, which has contaminated various military sites during its use, storage and manufacturing worldwide. As RDX is a recalcitrant, less soluble and toxic to human beings and other organisms, it is essential to remediate the contaminated sites. In the current investigation, authors have explored the potential of two indigenous microbes i.e., Bacillus toyonensis (isolate No. WS4-TSB-3, MTCC No. 12857) and Paenibacillus dendritiformis (isolate No. S10-TSA-3, MTCC No. 12859) isolated from an explosive manufacturing facility in north India, for the degradation of RDX in aqueous medium. Furthermore, RDX degradation has been optimized using response surface methodology (RSM) in a 15 days experiment at concentration of 20, 40, and 60 mg/L. It was found that various factors such as initial concentration of RDX, inoculum volume (2, 4 and 6%) and time (5, 10 and 15 days) had impact on transformation and degradation of contaminant. Samples were analyzed using high performance liquid chromatography (HPLC) and intermediate products were identified using LC-MS/MS. Maximum RDX removal of 81.6 ± 1.3 and 84.7 ± 0.9% for Bacillus toyonensis (isolate No. WS4-TSB-3) and Paenibacillus dendritiformis (isolate No. S10-TSA-3), respectively, was observed on 15th day at 40 mg/L initial concentration. During the degradation Hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), Hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), 4-Nitro-2,4-diazabutanal, Bis(hydroxymethyl)nitramine and nitrite were identified as intermediate products. The findings of the investigation suggest that both the microbes have the potential to degrade RDX in the aqueous medium and can be used for up-scaling the degradation of RDX on explosive contaminated sites.