Prebiotic Functions of Konjac Root Powder in Chocolate Milk Enriched with Free and Encapsulated Lactic Acid Bacteria.

This study investigated the prebiotic functions of Konjac root powder (KRP) when added to chocolate milk (ChM) enriched with 2% of free or microencapsulated lactic acid bacteria (FLAB or ELAB). The effects of different concentrations of KRP (0%, 2% and 4%) and refrigerated storage time on the physical, chemical and microbiological characteristics of this chocolate milk were examined. The results show that pH significantly declined (p < 0.05), while titratable acidity increased in all ChM samples with KRP and FLAB or ELAB during refrigerated storage. The pH values ranged from 6.0 ± 0.03 in samples enriched ELAB and 4% KRP to 6.33 ± 0.03 in ChM enriched with FLAB and 2% KRP. Viscosity of ChM was affected mainly by the added amounts of KRP and storage time. The largest viscosity (5500 cP) was observed in all samples containing 4% KPR on day zero and decreased significantly (p < 0.05) over storage time to reach 2800 cP in ChM samples containing 0% LAB and 4% KRP after 21 days of storage. Changes in LAB counts proved the initial hypothesis that KRP could act as prebiotics in the presence of LAB using chocolate milk as a carrier. The initial LAB counts in inoculated samples on day zero of refrigeration storage were not significantly different (p > 0.05) among all treatments. However, ChM enriched with 2% and 4% KRP and ELAB revealed significantly (p < 0.05) larger LAB counts (4.91 ± 0.78 and 5.0 ± 0.57 log CFU/mL, respectively) than the control (3.85 ± 0.55 log CFU/mL) after 21 days of storage.

Viability of Lactobacillus delbrueckii in chocolates during storage and in-vitro bioaccessibility of polyphenols and SCFAs.

This study evaluated the viability of encapsulated Lactobacillus delbrueckii subsp. bulgaricus in chocolate during storage and in-vitro gastrointestinal transit. Flavonoid contents and short chain fatty acids (SCFAs) production during gastrointestinal transit were also assessed. Encapsulated L. delbrueckii subsp. bulgaricus survived well in chocolates >7 logs both after 120 days of storage at 4 °C and 25 °C, and during in-vitro gastrointestinal transit. The release of SCFAs through in-vitro gastrointestinal digestion and colonic fermentation revealed that probiotic-chocolates could be an excellent source of nutrients for the gut microbiota. Encapsulated probiotic in chocolates with 70% cocoa produced significantly (P < 0.05) more acetic, propionic, isobutyric, butyric and isovaleric acids than that with 45% cocoa. The bioconversion results of a specific polyphenol by L. delbrueckii subsp. bulgaricus exhibited that chocolate polyphenols could be utilized by probiotics for their metabolism. These findings confirmed that chocolate could be successfully fortified with L. delbrueckii subsp. bulgaricus encapsulation to improve health promoting properties of chocolates.

The degradation of textile industry dyes using the effective bacterial consortium.

The effluents from textile industries without proper treatment contains a remarkable amount of synthetic dyes which are harmful to the environment and a big challenge globally to degrade it with a eco-friendly way. Conventional methods are extremely energy-consuming, non-effective and generate a toxic sludge impacting the environment. Several microorganisms can be utilized to treat these effluents. The research deals with five bacteria isolated from textile effluent and their consortium for the biodegradation ability of Novacron dyes. The isolates were identified through the Biolog™ identification system and molecular technique. Biodegradation was confirmed by measuring optical density (OD) optimizing conditions (pH 7.0, temperature 37 °C, 10 % inoculums and 100 mg/L dye) under static condition. The isolates started decolourization at 24 h whereas, the consortium started decolourization at 18 h and exhibited a maximum after 72 h. The presence of low molecular weight protein as metabolite supported the biodegradation and non hazardous to environment. This study revealed that these bacteria might have degradation potentials, and research results will help to set up dye removal eco-friendly methods to expose the dye effulents to environment in future.

Impact of encapsulating probiotics with cocoa powder on the viability of probiotics during chocolate processing, storage, and in vitro gastrointestinal digestion.

Chocolates can be formulated as a functional food via enrichment with probiotics. However, the added probiotics must overcome the challenges of processing and storage conditions and the harsh gastrointestinal environment. The study aimed to overcome these challenges using two different formulations of cocoa powder as alternative encapsulants along with Na-alginate (A1 ) and Na-alginate and fructooligosaccharides (A2 ). Seven different probiotic strains were encapsulated individually using the new formulations and viabilities of these encapsulated probiotics were assessed prior to and after they were added to chocolates. The highest achieved encapsulation efficiencies were 93.40% for formulation A1 (with Lactobacillus casei) and 95.36% for formulation A2 (with Lactobacillus acidophilus La5). The encapsulated probiotics with the new formulations maintained higher viability than the recommended therapeutic level (107 colony forming unit [CFU]/g) for up to 180 and 120 days of storage at 4 and 25 °C, respectively. The tested encapsulants improved probiotics survival when subjected to thermal stress and maintained about 9.0 Logs CFU/g at 60 °C. Additionally, the viable numbers of probiotics in fortified chocolates showed higher than 7 Logs CFU/g after 90 days of storage at 25 °C. Both formulations exhibited significantly (P < 0.05) high survivability of probiotics (8.0 Logs CFU/g) during the in vitro gastrointestinal digestion. This study demonstrated that cocoa powder along with Na-alginate and FOS has the potential to be used as a probiotic encapsulating material, and chocolates could be an excellent carrier for the development of healthy probiotic chocolate products. PRACTICAL APPLICATION: The introduction of cocoa powder as an effective encapsulating agent to deliver probiotics could help the chocolate industry to develop healthy and attractive functional snacks for health-conscious consumers.

The Prospects of Lactobacillus oris as a Potential Probiotic With Cholesterol-Reducing Property From Mother's Milk.

This experiment was conducted to characterize potential Lactobacillus spp. isolated from mother's milk and infant feces to obtain new and specific probiotic strains. In this study, seven ascendant strains were identified as Lactobacillus spp. based on their morphological characteristics and biochemical properties. Among them, only one (C-1) isolate was identified as Lactobacillus oris through BioLogTM identification. The study further investigated the isolate through probiotic potentiality tests such as pH and bile tolerance, NaCl tolerance test, gastric juice tolerance, antioxidant activity, resistance to hydrogen, reduction of sodium nitrate, antimicrobial activity, and antibiotic susceptibility test. The result showed that the strain is a potential probiotic based on probiotic capability. The identified strain was most acid-tolerant and retained around 80% viability for up to 4 h at pH 1.0 and 2.0. The isolate showed tolerance against up to 1.50% bile concentration and gastric juice and was able to grow 1-6% NaCl concentrations. Lactobacillus oris showed resistance to most antibiotics as well as antagonistic activity against the tested pathogen, good antioxidant properties, reduction of sodium nitrate and H2O2. The isolate exhibited good intestinal epithelial adhesion properties, and SDS page was performed for secreted protein analysis. Moreover, the strain showed promising cholesterol-lowering properties based on the cholesterol level. This present result indicates that L. oris has superior probiotic properties and can be regarded as a potential probiotic candidate.

Identification and Growth Characterization of a Novel Strain of Saccharomyces boulardii Isolated From Soya Paste.

The nonpathogenic yeast Saccharomyces boulardii (Sb) has beneficial effects on the human intestine, and thus has been prescribed as probiotics for the treatment of diarrhea and gastrointestinal diseases. This is the only commercialized yeast with the purpose of being used as human medicine. Currently, little is known about their multiple mechanisms of actions. The S. boulardii yeast strain is isolated and identified by using the BIOLOGTM microarray identification system and morphologically. To understand its functional roles, the present study investigates the ability of this yeast to tolerate different concentrations of bile salt up to 2.5%, cell hydrophobicity, antioxidants, autoaggregation activity, and simulated gastrointestinal digestion. The effect of temperatures (up to 50°C), pH (up to 8.0), and salinity (at best 7%) was also monitored on the growth and survival of the yeast cell. The physicochemical analyses revealed that S. boulardii could survive in stomach conditions at pH 2.5, temperature 37°C, and 2% bile salt. Antibiotic susceptibility of S. boulardii was carried out using commercial antibiotic discs. The antimicrobial activity of the isolated S. boulardii against bacterial pathogens related to diarrhea diseases was in-vitro determined by the Well Diffusion method. The biosafety assay findings also claimed S. boulardii could be a potential probiotic. The experimental findings suggest that the isolated S. boulardii possesses excellent probiotic capacities as a biotherapeutic agent for antidiarrheal and gastrointestinal disorders.

Evaluation of microplate immunocapture method for detection of Vibrio cholerae, Salmonella Typhi and Shigella flexneri from food.

BACKGROUND: Improved methods with better separation and concentration ability for detection of foodborne pathogens are in constant need. The aim of this study was to evaluate microplate immunocapture (IC) method for detection of Salmonella Typhi, Shigella flexneri and Vibrio cholerae from food samples to provide a better alternative to conventional culture based methods. RESULTS: The IC method was optimized for incubation time, bacterial concentration, and capture efficiency. 6 h incubation and log 6 CFU/ml cell concentration provided optimal results. The method was shown to be highly specific for the pathogens concerned. Capture efficiency (CE) was around 100% of the target pathogens, whereas CE was either zero or very low for non-target pathogens. The IC method also showed better pathogen detection ability at different concentrations of cells from artificially contaminated food samples in comparison with culture based methods. Performance parameter of the method was also comparable (Detection limit- 25 CFU/25 g; sensitivity 100%; specificity-96.8%; Accuracy-96.7%), even better than culture based methods (Detection limit- 125 CFU/25 g; sensitivity 95.9%; specificity-97%; Accuracy-96.2%). CONCLUSION: The IC method poses to be the potential to be used as a method of choice for detection of foodborne pathogens in routine laboratory practice after proper validation.

Formulation and nutritional evaluation of a healthy vegetable soup powder supplemented with soy flour, mushroom, and moringa leaf.

The research study was conducted to develop a healthy vegetables soup powder supplemented with soy flour, mushroom, moringa leaf and compare its nutritional facts with locally available soup powders. Proximate analysis and sensory evaluation were done by standard method. In this study, moisture, ash, protein, fat, fiber, carbohydrate, and energy content were ranged from 2.83% to 5.46%, 9.39% to 16.48%, 6.92% to 16.05%, 4.22% to 6.39%, 0.22% to 1.61%, 58.81% to 75.41%, and 337.42 to 386.72 kcal/100 g, respectively. Highest content of vitamin D, minerals, protein, and fiber and lowest content of moisture, fat, and carbohydrate were found in the presently developed soy-mushroom-moringa soup powder compare to locally available soup powders. Vitamin C was also found significantly higher than locally available soup powders S1, S2, and S3. Heavy metals were not found in any of the soup powders. On the sensory and microbiological point of view, the presently developed soup powder was found highly acceptable up to 6 months. So, the developed soy-mushroom-moringa soup powder is nutritionally superior to locally available soup powders and sufficient to meet day-to-day nutritional requirements as a supplement.

Chemical composition and pharmacological significance of Anethum Sowa L. Root.

BACKGROUND: Medicinal herbs are used for the treatment of different ailments since antiquity. Different parts of Anethum sowa L. is used in folk medicine as a carminative for the treatment of flatulence, colic and hiccups of infants and children, antioxidant, antimicrobial and antispasmodic agent. The aim of our present study is to evaluate the chemical composition of the essential oil, proximate and elemental composition, amino acid, fatty acid profile and thermal behaviour of its root part as well as different pharmacological activities like antioxidant, antimicrobial and cytotoxicity of the root essential oil. METHODS: The air-dried roots of Anethum sowa L. were subjected to hydro-distillation to yield the essential oil. The antioxidant activity of the essential oil was studied by DPPH radical scavenging activity. The antimicrobial activity was tested against four Gram-positive, six Gram-negative bacteria and four fungi species. The minimum inhibitory concentration (MIC) and Minimum bacterial concentration (MBC) for each examined microorganism were determined using the micro-dilution method. The LC50 value of the oil was also evaluated by brine shrimp lethality assay. The subsequent proximate analysis was also done by AOAC methods. The elemental analysis of the root powder was analysed by ICP-MS, AAS and FP system. The fatty acid was extracted by hot and cold extraction method and the analyses were carried out by GC. The amino acid profile was done by the amino acid analyzer. The DTA, DTG and TG of the root powder were taken by the thermogravimetric analyzer. RESULTS: A total of 24 constituents was identified and quantified in the essential oil and its water extract portion by GC and GC-MS. Apiol (81.99 and 74.779%) was found the highest phenylpropanoid constituent followed by m-diaminobenzene (10.446 and 8.778%) in the essential oil and aqueous extract portion. On the other hand, ß-butyrolactone (5.13%) and isobutyl acetone (3.73%) were found in the major constituents in the water extract part. The IC50 value of the essential oil was found to be 3.07 mg/mL by DPPH radical assay methods. The LC50 value of the brine shrimp cytotoxicity assay of the essential oil was observed at 0.81 µg/mL. The essential oil showed better activity on Gram-negative bacteria than Gram-positive bacteria and fungi. The proximate composition showed that root contained 5.29% ash, 2.01% protein, 54.09% crude fibre, 0.15% essential oil and 1.14% fatty oil for hot extract and 0.23% for cold extract on the dried basis. The palmitic (33.81 & 31.58%) and linoleic acid (30.03 & 23.79%) were the major saturated and unsaturated fatty acids in the cold and hot extracted root powder respectively. Ca (23,600 mg/kg), Mg (7620.33 mg/kg) and K (1286.15 mg/kg) were the most predominant elements followed by Ni (1187.30 mg/kg), Se (913.79 mg/kg), Li (317.84 mg/kg), Na (288.72 mg/kg) and Fe (206.88 mg/kg). The toxic elements were found to be within the permissible limit. Glutamic acid (19.37%), glycine (14.53%) and lysine (17.08%) were found as the major amino acids. The decomposition rates were obtained by TG, DTG and DTA curve of the powder sample at various temperature ranges. CONCLUSIONS: The results demonstrated that the root part of Anethum sowa L. is a rich source of mineral elements, essential amino acid and fatty acids. The essential oil is the highly potential as bioactive oil for pharmaceuticals and medical applications, possessing antioxidant, antimicrobial and cytotoxic activities. The thermal analysis suggested as a simple, effective and rapid method to characterize the Anethum sowa L. species as well as to assess for herbal formulation.

Antimicrobial and antioxidant activities of Saccharomyces cerevisiae IFST062013, a potential probiotic.

BACKGROUND: Probiotic yeast has become a field of interest to scientists in recent years. METHODS: Conventional cultural method was employed to isolate and identify yeast and standard methods were used to determine different probiotic attributes, antimicrobial and antioxidant properties. RESULTS: This study reports potential probiotic properties of a strain of S. cerevisiae IFST 062013 isolated from fruit. The isolate is tolerant to a wide range of temperature and pH, high concentration of bile salt and NaCl, gastric juice, intestinal environment, α-amylase, trypsin and lysozyme. It can produce organic acid and showed resistance against tetracycline, ampicillin, gentamycin, penicillin, polymixin B and nalidixic acid. It can assimilate cholesterol, can produce killer toxin, vitamin B12, glutathione, siderophore and strong biofilm. It showed moderate auto-aggregation ability and cell surface hydrophobicity. The isolate can produce enzymes such as amylase, protease, lipase, cellulose, but unable to produce galactosidase. The isolate can't produce gelatinase and DNase. The isolate showed moderate anti-microbial activity against bacteria and fungi and cell lysate showed better antimicrobial activity than whole cell and culture supernatant. Again, the isolate showed better anti-bacterial activity against gram negative bacteria than gram positive. The isolate showed strong antioxidant activity, reducing power, nitric oxide and hydroxyl radical scavenging activity, significant brine shrimp cytotoxicity and acute toxicity and metal ion chelating activity. The isolate did not induce any detectable change in general health of mice upon oral toxicity testing and found to be safe in mouse model. The isolate improve lymphocyte proliferation and cytokine production in treated mice. CONCLUSIONS: Such isolate could be potential as probiotic to be used therapeutically.

Pharmacological effects of the phytochemicals of Anethum sowa L. root extracts.

BACKGROUND: Anethum sowa L. is widely used as an important spice and traditional medicinal plants to treat various ailments. On the basis of scientific ethnobotanical information, this study was undertaken to evaluate the antioxidant, antimicrobial and cytotoxic activity of the crude extracts of Anethum sowa L. roots as well as to identify the classes of phytochemicals by chemical tests. METHODS: The antioxidant potential of the extracts was ascertained with the stable organic free radical (2, 2-diphenyl-1-picryl-hydrazyl). The agar well diffusion method was used to determine the susceptibility of bacterial and fungal strains of the crude extracts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentrations (MBC) were determined by the microdilution test. Cytotoxic activities were screened using brine shrimps (Artemia salina) lethality assay. Finally, phytochemicals were profiled using standard procedures. RESULTS: A preliminary phytochemical screening of the different crude extracts by methanol, ethyl acetate and chloroform showed the presence of secondary metabolites such as flavonoids, alkaloids, saponin, cardiac glycosides and tannins while cyanogenetic glycosides were not detected. The methanol, ethyl acetate and chloroform extracts displayed high antioxidant activity (IC50 = 13.08 ± 0.03, 33.48 ± 0.16 and 36.42 ± 0.41 µg/mL, respectively) in the DPPH assay comparable to that of the standard ascorbic acid and BHT (IC50 = 3.74 ± 0.05 and 11.84 ± 0.29 µg/mL). The cytotoxic activity of the crude ethyl acetate and chloroform extracts possessed excellent activity (LC50 = 5.03 ± 0.08, 5.23 ± 0.11 and 17.22 ± 0.14 µg/mL, respectively) against brine shrimp larvae after 24 h of treatment and compared with standard vincristine sulphate (LC50 = 0.46 ± 0.05 µg/mL). The extracts also showed good antimicrobial activity against both Gram-positive and Gram-negative bacteria when compared with two standard antibiotics ciprofloxacin and tetracycline. CONCLUSION: These results showed that the Anethum sowa root extracts are the important source of the antioxidant, antimicrobial and cytotoxic agent. So, further research is necessary to isolate and characterize of different phytoconstituents for pharmaceutical drug lead molecules and also to verify its traditional uses.

Characterization of aflatoxin producing Aspergillus flavus from food and feed samples.

Aspergillus flavus is one of the major producers of aflatoxin and can contaminate wide range of agricultural commodities either in field or in storage. 15 presumptive Aspergillus flavus has been isolated from 30 feed and grain samples. All the isolates were morphologically similar to Aspergillus flavus type strains. All the isolates were found to be aflatoxigenic. DNA sequencing of 5.8 s rDNA confirmed all of them to be Aspergillus flavus. Only 1 isolate possessed all the seven toxigenic gene (aflR, aflS, aflQ, aflP, aflD, aflM, and aflO) while aflP & aflQ were most prevalent in the isolates. All the isolates possessed at least three toxigenic genes. Toxin producing ability in solid culture media showed that 11 isolates isolates were able to produce both aflatoxin B1 & B2. More than 90% isolates produced aflatoxin B1 ranging 7-22 µg/g of agar. This study alarms us about the potential risks of Aspergillus flavus to public health if contaminate agricultural commodities such as grains or raw materials such as poultry feed. Proper harvest and storage management is required to reduce the risk of aflatoxin in feed and grains.

Minimum ignition energy of nano and micro Ti powder in the presence of inert nano TiO2 powder.

The inerting effect of nano-sized TiO2 powder on ignition sensitivity of nano and micro Ti powders was investigated with a Mike 3 apparatus. "A little is not good enough" is also suitable for micro Ti powders mixed with nano-sized solid inertants. MIE of the mixtures did not significantly increase until the TiO2 percentage exceeded 50%. Nano-sized TiO2 powders were ineffective as an inertant when mixed with nano Ti powders, especially at higher dust loadings. Even with 90% nano TiO2 powder, mixtures still showed high ignition sensitivity because the statistic energy was as low as 2.1 mJ. Layer fires induced by ignited but unburned metal particles may occur for micro Ti powders mixed with nano TiO2 powders following a low level dust explosion. Such layer fires could lead to a violent dust explosion after a second dispersion. Thus, additional attention is needed to prevent metallic layer fires even where electric spark potential is low. In the case of nano Ti powder, no layer fires were observed because of less flammable material involved in the mixtures investigated, and faster flame propagation in nanoparticle clouds.

Minimum ignition temperature of nano and micro Ti powder clouds in the presence of inert nano TiO2 powder.

Minimum ignition temperature (MIT) of micro Ti powder increased gradually with increases in nano-sized TiO2 employed as an inertant. Solid TiO2 inertant significantly reduced ignition hazard of micro Ti powder in contact with hot surfaces. The MIT of nano Ti powder remained low (583 K), however, even with 90% TiO2. The MIT of micro Ti powder, when mixed with nano Ti powder at concentrations as low as 10%, decreased so dramatically that its application as a solid fuel may be possible. A simple MIT model was proposed for aggregate particle size estimation and better understanding of the inerting effect of nano TiO2 on MIT. Estimated particle size was 1.46-1.51 µm larger than that in the 20-L sphere due to poor dispersion in the BAM oven. Calculated MITs were lower than corresponding empirically determined values for micro Ti powder because nano-sized TiO2 coated the micro Ti powder, thereby decreasing its reaction kinetics. In the case of nano Ti powder, nano-sized TiO2 facilitated dispersion of nano Ti powder which resulted in a calculated MIT that was greater than the experimentally determined value.

Nucleic acid amplification: Alternative methods of polymerase chain reaction.

Nucleic acid amplification is a valuable molecular tool not only in basic research but also in application oriented fields, such as clinical medicine development, infectious diseases diagnosis, gene cloning and industrial quality control. A comperehensive review of the literature on the principles, applications, challenges and prospects of different alternative methods of polymerase chain reaction (PCR) was performed. PCR was the first nucleic acid amplification method. With the advancement of research, a no of alternative nucleic acid amplification methods has been developed such as loop mediated isothermal amplification, nucleic acid sequence based amplification, strand displacement amplification, multiple displacement amplification. Most of the alternative methods are isothermal obviating the need for thermal cyclers. Though principles of most of the alternate methods are relatively complex than that of PCR, they offer better applicability and sensitivity in cases where PCR has limitations. Most of the alternate methods still have to prove themselves through extensive validation studies and are not available in commercial form; they pose the potentiality to be used as replacements of PCR. Continuous research is going on in different parts of the world to make these methods viable technically and economically.

Critical Factors Affecting the Success of Cloning, Expression, and Mass Production of Enzymes by Recombinant E. coli.

E. coli is the most frequently used host for production of enzymes and other proteins by recombinant DNA technology. E. coli is preferable for its relative simplicity, inexpensive and fast high-density cultivation, well-known genetics, and large number of compatible molecular tools available. Despite all these advantages, expression and production of recombinant enzymes are not always successful and often result in insoluble and nonfunctional proteins. There are many factors that affect the success of cloning, expression, and mass production of enzymes by recombinant E. coli. In this paper, these critical factors and approaches to overcome these obstacles are summarized focusing controlled expression of target protein/enzyme in an unmodified form at industrial level.

Evaluation of health risk and arsenic levels in vegetables sold in markets of Dhaka (Bangladesh) and Salamanca (Spain) by hydride generation atomic absorption spectroscopy.

The main objective of this study was to determine arsenic (As) levels in vegetables collected from the markets of Dhaka, Bangladesh and for comparison from Salamanca, Spain by HGAAS under optimal conditions, and the potential health risk from consumption of these vegetables. The mean and range of the total As concentration in the vegetables from the markets of Dhaka, Bangladesh were 114 and 1.0-293 µg/kg, respectively. Total As concentration in 77% of Bangladesh vegetables measured was higher than that recommended by the standard. The mean and range of As concentrations for vegetables grown in Spain were 65 and bdl-130 µg/kg, respectively, for Salamanca, 102 and bdl-423 µg/kg, respectively, for Almeria. The As content of the Bangladesh vegetables was approximately twofold to threefold higher than those observed for the vegetables from Almeria and Salamanca (Spain), but in some cases, were similar or less. Daily consumption of As-rich vegetables may result in an additional source of As in the diet, based on the provisional tolerable intake of As for adults recommended by WHO.