Effect of enzyme pretreatment in the ultrasound assisted extraction of finger millet polyphenols.

Polyphenols from plant sources are increasingly recognized as functional ingredients with multiple health benefits. Finger millet (Eleusine coracana), a major millet grown in Asia and Africa is a significant source of polyphenols. The extraction of polyphenols from millets which are under-utilized has received less research attention compared to other plant matrices. Therefore, the present study was carried out to identify an effective method of extracting the polyphenols from finger millet (v. GPU 28) seed coat (FMSC). Two eco friendly methods namely ultrasonication (UA) and enzyme treatment followed by ultrasonication (EUA) were compared with the conventional heat reflux method (HR). The polyphenolic profile and content along with the antioxidant potential of the extract were evaluated. Phenolic yield increased 2.3 fold using EUA with xylanase (XUA) compared to heat reflux extraction (HR). However, yield with UA was equivalent to the conventional method. Total flavonoids increased 1.4 fold in UA and 1.3 fold in XUA, similarly, tannins also showed a significant increase (1.1 fold in UA and 1.2 fold in XUA). FTIR spectra revealed the presence of all phenolic functional groups and ESI-MS showed 80% similarity in the individual polyphenols in the extracts. Catechins, luteolin and cyanidin were identified in the UA and XUA treated samples, whereas, shikimic acid derivatives- caffeoyl and di caffeoyl were present only in XUA extracts. This is the first report on enhanced extraction of polyphenols from FMSC using a combination of enzyme treatment and ultrasonication, providing a green technology for utilization of polyphenols in nutraceuticals and functional foods.

Determination of forced convection heat transfer coefficients and development of empirical correlations for milk in vessel with mechanical agitators.

Heating of milk is an important unit operation to produce many milk based products. This process is done either in heat exchangers or in agitated vessels, where the mode and type of heat transfer plays a significant role. Use of mechanical agitator with suitable impeller would result in uniform agitation of the liquid. Data for forced convection heat transfer coefficients for milk in agitated vessel have not been documented. In the present investigation, forced convection heat transfer coefficients for milk, using mechanical agitators in vessels have been determined. Experiments have been conducted in vessels (Baffled as well as Unbaffled vessel) that were designed and fabricated under standard procedure employing: (1) Flat Six Blade Turbine impeller, (2) Inclined Six Blade Turbine impeller, (3) Three Blade Propeller impeller and (4) Two Bladed Paddle. Heat transfer coefficients were determined for h¯ for cow milk (3.5% fat), standardised milk (4.5% fat) and full cream milk (6.0% fat). The Two Bladed Paddle was found to as yield the highest heat transfer coefficient h¯ for all three types of milk. In addition, empirical correlations for the forced convection heat transfer using Nu=a·Reb·Pr0.33 have been developed for each case. These heat transfer correlations derived will find application in large scale design and the experimental setup would be useful for future investigation with different liquids and impellers.

Accelerated fermentation of Idli batter using Eleusine coracana and Pennisetum glaucum.

The present study focuses on reduction in fermentation time of idli batter and its enrichment with calcium and iron using finger millet and pearl millet. Fermentation time decreased from 12 to 6 h with addition of finger millet and to 8 h with pearl millet. Rate of fermentation, bulk density, viscosity, microbial changes and particle morphology were assessed in the batter; nutritional quality, sensory, phytochemicals and internal characteristics of idli were investigated. The color and texture characteristics of both batter and idli were evaluated. The pH and titratable acidity of control batter at 6 h was 5.32 and 0.27%, addition of finger and pearl millet changed to 4.32, 0.45% and 4.53, 0.45% at respective fermentation times (6 and 8 h). The viable yeast, lactic acid bacteria and total bacterial count (log CFU g-1) in the batters increased with time, reaching 1.26, 3.85, 4.56 (control); 2.32, 9.84, 9.58 (finger millet) and 1.76, 7.34, 7.74 (pearl millet) respectively at the end of 6 h of fermentation. Addition of finger millet and pearl millet flour (10% w/w) to the batter enhanced the dietary fiber by 28 and 23%, calcium by 113 and 56%, iron by 51 and 258% in the respective idlies when compared with the control.

Evaluation of antimicrobial activity and probiotic properties of wild-strain Pichia kudriavzevii isolated from frozen idli batter.

The present research was undertaken to study the probiotic characteristics of Pichia kudriavzevii isolated from frozen idli batter. Polymerase chain reaction amplification with 18S rRNA primers confirmed Pichia kudriavzevii, a xylose-utilizing probiotic strain. It was resistant to physiological concentrations of bile salts, pepsin and pancreatic enzyme. It also showed efficient auto-aggregation as well as co-aggregation ability with four commercial probiotic yeasts and exhibited good hydrophobicity in xylene and toluene. The strain inhibited the growth of 13 enteropathogens and showed a commensal relationship with four commercial probiotic yeast and bacteria. Moreover, it was resistant to 30 antibiotics with different modes of action. The yeast exhibited thermotolerance up to 95 °C for 2 h. The cell-free supernatants were also found to be heat stable, indicating the presence of thermostable secondary metabolites. Hence it could be exploited as starter culture, co-culture or probiotic in the preparation of fermented products or incorporated in heatable foods as well. Copyright © 2016 John Wiley & Sons, Ltd.

Functional characterization and microencapsulation of probiotic bacteria from koozh.

Koozh is a traditional fermented millet beverage unique to south India. Analysis of six market samples of koozh for their microbial profile resulted in 69 isolates of presumptive lactic acid bacteria (LAB). They were grouped as Leuconostoc sp., Enterococcus sp., Streptococcus sp. and Lactobacillus sp. based on morphological characteristics and biochemical tests. Eight among them showed probiotic features: resistance to acid (2.5 pH for 6 h), resistance to 0.3 % ox bile, moderate hydrophobicity (40 %), antibacterial activity against 10 pathogens, susceptibility to 50 % of antibiotics tested. Sequencing of 16srDNA showed them to be five strains of Enterococcus hirae and one each of Enterococcus facecalis, Bacillus amyloliquefaciens and Lactobacillus plantarum. The probiotic isolates were encapsulated in skim milk powder by two different drying techniques: freeze drying and spray drying. The encapsulated probiotic isolates survived both in simulated gastric fluid and simulated intestinal fluid with high cell viability (98-99 %). Storage for 16 weeks at room temperature (27 °C), resulted in 2 log reduction, but better survival with only 1 log reduction was observed at 4 °C and was best at -20 °C. Survival of isolates was similar in both spray and freeze dried products.

Effect of pulsed electric field (PEF) treatment on sugarcane juice.

This study was carried out to evaluate the effect of PEF process using static treatment chamber on fresh sugarcane juice with and without addition of lemon and ginger with respect to microbial content, chemical properties, nutrient content and shelf life extention of the product. The fresh sugar cane juice without addition of lemon and ginger treated at different field strengths (30 kV cm-1 and 50 kV cm-1) and different pulse numbers (150, 300) was initially investigated by storage at room temperature (31 °C) and refrigeration temperature (4 °C) for 30 days. The PEF effect on fresh sugar cane juice at room temperature and refrigerated temperature was compared with untreated sample (31 °C). At the end of the storage period samples treated at field strength 30 kV cm-1, 150 pulses were found to be stable compared with untreated sample. The second experimental study of PEF process was done on fresh sugarcane juice with the addition of lemon and ginger for fourteen days at different electric field intensities (10 kV cm-1, 20 kV cm-1 and 30 kV cm-1) with the same pulse number (150 pulses) and stored 4 °C. Even better reduction of microbes was achieved with PEF treatment condition of field strength 20 kV cm-1, 150 pulses in the presence of lemon and ginger. The sensory attributes of untreated fresh sugarcane juice were maintained up to only two days, but for the PEF treated sample, shelf life was extened up to seven days. Further, addition of lemon and ginger in the PEF treated sugarcane juice doubled the shelf life up to fourteen days.

Fructan distribution in banana cultivars and effect of ripening and processing on Nendran banana.

Many plants store fructan as reserve carbohydrate. Fructans naturally present in almost all plant foods, are also used as functional ingredients by the food industry to modify the texture and taste due to their properties as gelling agents, fat substitutes, soluble dietary fibers and low calorie sweeteners. Seven banana cultivars were analysed for fructans and Nendran banana was selected for the next set of experiments as it had the highest fructan content (1433.3 mg/100 g) among the cultivars studied. Low temperature ripening (16 °C) of Nendran banana resulted in higher fructan accumulation of these carbohydrates in cold conditions. Pectinase pre-treatment significantly increased yield of total fructans from 1.4/100 g to 6.5 g/100 g i.e., 370 %. Fructan composition was affected by processing, namely steaming and puree preparation in Nendran. The fructan composition data documented in this study will enable including banana, naturally high in fructans in the diet and will facilitate storage and processing for nutritional formulation for higher fructan consumption.

Probiotic and Functional Characterization of Pediococcus acidilactici Isolated from Bhaati jaanr, Traditional Fermented Rice Porridge.

Traditional fermented foods are the ideal source of novel probiotic isolates which are known to have significant therapeutic benefits and play a vital role as bioprotective agents. Bhaati jaanr is an ethnic fermented rice beverage popularly consumed in sub-Himalayan regions. The strain UAMS was isolated from Bhaati jaanr based on high butyrate production and evaluated for the potential probiotic characteristics. MALDI-TOF MS and 16 s rRNA gene sequencing revealed the identity of strains as Pediococcus acidilactici. The isolated strain exhibited high tolerance to gastric and bile stress, autoaggregation, hydrophobicity, and adherence to colon cells. Antibiotic susceptibility testing results showed the resistance of the isolated strain toward tested common antibiotics and the pathogenic determinants were absent in PCR-based detection. Moreover, the organism was able to inhibit the growth of Listeria, Salmonella, Staphylococcus, and Enterococcus species. The isolate was found to be a high butyrate producer along with other short-chain fatty acids and exhibited an anti-proliferative effect against colon cancer cells HT29 and SW480. Therefore, our study represents Pediococcus acidilactici UAMS as a potent putative probiotic with bioprotective abilities.

In Vitro Probitotic Evaluation of Saccharomyces boulardii with Antimicrobial Spectrum in a Caenorhabditis elegans Model.

In the present study, we screened for potential probiotic yeast that could survive under extreme frozen conditions. The antimicrobial and heat-stable properties of the isolated yeast strains Saccharomyces boulardii (S. boulardii) (KT000032, KT000033, KT000034, KT000035, KT000036, and KT000037) was analyzed and compared with commercial probiotic strains. The results revealed that the tested S. boulardii KT000032 strain showed higher resistance to gastric enzymes (bile salts, pepsin, and pancreatic enzyme) at low pH, with broad antibiotic resistance. In addition, the strain also showed efficient auto-aggregation and co-aggregation abilities and efficient hydrophobicity in the in-vitro and in-vivo C. elegens gut model. Further, the KT000032 strain showed higher antimicrobial efficiency against 13 different enteropathogens and exhibited commensal relationships with five commercial probiotic strains. Besides, the bioactive compounds produced in the cell-free supernatant of probiotic yeast showed thermo-tolerance (95 °C for two hours). Furthermore, the thermo-stable property of the strains will facilitate their incorporation into ready-to-eat food products under extreme food processing conditions.

Effect of Rice Processing towards Lower Rapidly Available Glucose (RAG) Favors Idli, a South Indian Fermented Food Suitable for Diabetic Patients.

The Asian food pattern primarily embraces rice and rice-based products, which mainly comprise 90% starch. Among these various food products, Idli is mostly prepared through fermentation. It has high palatability, and the rapid and highly digestible nature of the food product results in a higher post-glucose level in diabetic patients. A heat-treated Idli rice sample was prepared by roasting parboiled rice at the temperature range of 155 to 165 °C for 65 to 75 s. Idli/rice-based Dokala made from heat-treated rice is better when compared to untreated rice in terms of its microbiological profile and physiochemical properties. The proximate composition of heat-treated parboiled rice Idli/Rice Dokala showed slightly higher values than the untreated parboiled rice Idli/Rice Dokala, which reveals that it has marginally higher nutritive value. Determination of the Rapidly Available Glucose (RAG) and Slowly Available Glucose (SAG) values, SEM analysis, resistant starch analysis, microbial assay, and in vivo studies were performed to determine the glycemic index (GI) and glycemic load in normal and diabetic persons. Sensory analysis also proved that heat-treated Idli/Rice Dokala is superior to untreated based on the color, flavor, appearance, taste, and texture.

Chemical, microbial and antibiotic susceptibility analyses of groundwater after a major flood event in Chennai.

During floods, human exposure to pathogens through contaminated water leads to the outbreak of epidemic diseases. This research presents the first extensive assessment of surface and groundwater samples collected immediately after a flood (December 2015) and post-flood (April 2016) from the Adyar River of Chennai, a major city in India, for major ions, trace metals, bacterial population, and pathogens. Severe rains in a short period of time resulted in flooding which inundated the wells, allowing the entry of sewage contaminated river water into the groundwater zone. This has led to bacterial counts and chemical ions exceeding Bureau of Indian Standard's recommended limits in most flood affected areas. Pathogens isolated from the groundwater showed resistance to antibiotics, namely ceftriaxone, doxycycline and nalidixic acid. However, they were sensitive to chloramphenicol, ciprofloxacin, norfloxacin, and tetracycline. Determining the antibiotic susceptibility of pathogens will help in the treatment of humans affected by contaminated water through an appropriate selection of prescribed medication.

Physicochemical properties of cookies enriched with xylooligosaccharides.

The growing commercial importance of xylooligosaccharides is based on their beneficial health properties, particularly their ability to stimulate the growth and activity of intestinal bacteria such as Bifidobacterium and Lactobacillus species. Xylooligosaccharides are less sweet, acid, and heat stable, with low recommended levels of intake compared to other oligosaccharides. In view of the consumer demand for foods with low sugar, low fat, and high fiber contents, they are suitable for incorporation into bakery products. In this study, we have developed wheat-based cookies incorporated with xylooligosaccharides at 5%, 10%, and 15% levels. The nutritive value and physicochemical properties of the cookies changed with xylooligosaccharides incorporation; both crude fiber and dietary fiber contents increased by 14% and 35%, respectively, in the enriched cookies. The moisture levels increased with increase in the percentage of xylooligosaccharides incorporated. Cookies with 5% xylooligosaccharides were found most acceptable, although the color was slightly darker compared to the control, while cookies with 10% and 15% xylooligosaccharides were softer and darker and therefore less acceptable. Enrichment with xylooligosaccharides at 5% provided a product stable for 21 days at room temperature (25 ± 2℃). The storage stability of cookies with higher levels of xylooligosaccharides was less than the 5% xylooligosaccharides cookies and control. The retention of the prebiotic xylooligosaccharides in the products was relatively high (74%).

DNA vaccination with VP2 gene fragment confers protection against Infectious Bursal Disease Virus in chickens.

Infectious Bursal Disease Virus (IBDV) causes immunosuppression in young chickens by destruction of antibody producing B cells in the Bursa of Fabricius and poses a potential threat to the poultry industry. We have examined the protective efficacy of a subunit DNA vaccine against IBDV infection in chickens in this study. An immunodominant VP2 gene fragment (VP252-417) was cloned into CMV promoter based DNA vaccine vector pVAX1 and in vitro expression of the DNA encoded antigens was confirmed by transfection of CHO cells with vaccine constructs followed by RT-PCR and western blot analysis using IBDV-antiserum. Two weeks old chickens were immunized intramuscularly with pVAXVP252-417 and the in vivo transcription of the plasmid DNA was confirmed by RT-PCR analysis of DNA injected muscle tissue at different intervals of post immunization. Tissue distribution analysis revealed that the plasmid DNA was extensively distributed in muscle, spleen, kidney, liver, and bursa tissues. Chickens immunized with pVAXVP252-417 developed high titer (1:12,000) of anti-VP252-417 antibodies. Further, chicken splenocytes from pVAXVP252-417 immunized group showed a significantly high proliferation to the whole viral and recombinant antigen (P<0.01) compared to control groups, which implies that pVAXVP252-417 codes for immunogenic fragment which has epitopes capable of eliciting both B and T cell responses. This is evident by the fact that, pVAXVP252-417 immunized chicken conferred 75% protection against virulent IBDV (vIBDV) challenge compared to the control group. Thus, the present study confirms that the immunodominant VP2 fragment can be used as a potential DNA vaccine against IBDV infection in chickens.

Protective immune responses of recombinant VP2 subunit antigen of infectious bursal disease virus in chickens.

Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease and poses a huge threat to poultry industry. The risks associated with conventional attenuated viral vaccines make it indispensable to probe into the development of novel and rationally designed subunit vaccines which are safer as well as effective. VP2 is the major host-protective antigen found in IBDV capsid. It encompasses different independent epitopes responsible for the induction of neutralizing antibody. Here, we report the efficacy of the immunodominant fragment of VP2 which induces both humoral and cellular immunity against infectious bursal disease. A 366 bp fragment (52-417 bp) of the VP2 gene from an IBDV field isolate was amplified and expressed in Escherichia coli as a 21 kDa recombinant protein. The efficacy of rVP2(52-417) antigen was compared with two commercial IBDV whole virus vaccine strains. The rVP2(52-417) induced significantly high antibody titres in chicken compared to commercial vaccines and the anti-rVP2(52-417) sera showed reactivity with viral antigens from both commercial strains (P<0.0001) and field isolates. Also, the chicken splenocytes from rVP2(52-417) immunized group showed a significantly high proliferation (P<0.01) compared to other groups, which implies that the rVP2(52-417) fragment contains immunogenic epitopes capable of eliciting both B and T cell responses. Further, rVP2(52-417) conferred 100% protection against vIBDV challenge in the immunized chickens which was significantly higher (P<0.001) compared to 55-60% protection by commercial vaccine strains. Hence, the study confirms the efficacy of the immunodominant VP2 fragment that could be used as a potent vaccine against IBDV infection in chicken.