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Anand Agricultural University, Anand
Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur
AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.
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ThesisItem Open Access IN-VITRO EVALUATION OF PROBIOTICS ON ORAL HEALTH(DEPARTMENT OF DAIRY MICROBIOLOGY SHETH M.C. COLLEGE OF DAIRY SCIENCE ANAND AGRICULTURAL UNIVERSITY ANAND, 2018) Shilpa Sasikumar Nair; Dr. J. B. PrajapatiProbiotics are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. In the last few decades, there is an increased demand worldwide for the products containing probiotic. Recently, the potential application of probiotics to maintain oral health and treat oral diseases has gained attention of several researchers. Although only few clinical studies have been conducted, the results to date suggests that the probiotics and its products can not only be used to prevent and treat oral infections but also to maintain the overall oral health. Considering this fact, the present study was planned to evaluate five potential probiotic strains for oral health by in-vitro tests and by developing model dental cream.ThesisItem Open Access FUNCTIONAL EVALUATION OF PROBIOTIC FERMENTED MILK ENRICHED WITH FINGER MILLET (ELEUSINE CORACANA) BY IN VITRO AND IN VIVO METHODS(DEPARTMENT OF DAIRY MICROBIOLOGY SHETH M.C. COLLEGE OF DAIRY SCIENCE ANAND AGRICULTURAL UNIVERSITY ANAND, 2018) Chaudhari Jinalben Kesharbhai; Dr. Sreeja V.Consumer interest in functional foods is increasing world over due to the vital role such foods play in the prevention and management of various diseases including Diabetes Mellitus (DM). The prevalence of DM, especially type 2 is increasing at an alarming rate worldwide. The side effects of existing treatments of DM are many. The major determinants of DM are said to be excess body fat, poor diet, physical inactivity, high blood pressure, and family history of diabetes. Diet strategies are considered one of the most appropriate prevention and management strategies for T2DM. Hence it becomes imperative that the protective role of functional foods in the prevention and management of DM should be investigated and communicated to the consumers. Probiotic fermented dairy products are said to play a vital role in the prevention and management of DM. Finger millets having high dietary fibre and phenolic content is considered beneficial for diabetic patients. Finger millet malting and fermentation processes decreases its antinutritional factors, improve the carbohydrate digestibility and glycemic response. Hence, the resultant product of milk-millet fermentation using probiotics may play a vital role in the prevention and management of DM. Very scanty research works are available on fermented products prepared through milk-finger millet fermentation and the functional evaluation of resultant product such as probiotic fermented milk enriched with finger millet. Hence, the present study was carried out to evaluate the probiotic fermented milk enriched with finger millet for its sensory, physico-chemical, compositional, and microbial parameters as well as functional properties by in vitro methods and antidiabetic activity by in vivo animal study.ThesisItem Open Access ISOLATION AND PURIFICATION OF ACEINHIBITORY PEPTIDES DERIVED FROM FERMENTED SURTI GOAT MILK(DEPARTMENT OF DAIRY MICROBIOLOGY SHETH M. C. COLLEGE OF DAIRY SCIENCE ANAND AGRICULTURAL UNIVERSITY ANAND, 2017) Parmar Heena Premjibhai; Dr. Subrota HatiFermented goat milk has multiple therapeutic and nutritional effects. Goat milk has lot of health benefits like antihypertensive, antioxidant and antimicrobial activity. But there is scanty information on ACE-inhibitory activity of fermented Surti goat milk (Indian breed). The present study was formulated to isolate and purify the ACE-inhibitory peptides from fermented goat milk (Capra aegagrus hircus) with a specific sequence of amino acids having ACE-inhibitory activity (in vitro).ThesisItem Open Access DEVELOPMENT OF FINGER MILLET (ELEUSINE CORACANA) ENRICHED PROBIOTIC FERMENTED MILK PRODUCT(DEPARTMENT OF DAIRY MICROBIOLOGY SHETH M.C. COLLEGE OF DAIRY SCIENCE ANAND AGRICULTURAL UNIVERSITY ANAND, 2017) SHAIKH AIJAZ SHAIKH MOHAMMAD; Dr. SREEJA V.Milk products are considered excellent media to generate an array of products that fit to current consumer demand for health benefitting foods. Fermented dairy products enriched with probiotic bacteria have developed into one of the most successful category of functional foods and the market of such probiotic dairy foods is increasing annually. Even though milk is considered as an almost complete food, it is deficient in dietary fibre, micronutrients such as iron and vitamin C. Finger millet or Ragi (Eleusine corcana) is one of the common millets in several regions of India. This millet is exceptionally rich in calcium, phosphorus and contains iron and many other trace elements and vitamins. Ragi is a good source of dietary fibre also. Additionally it is said to possess a number of health benefitting properties. Very few research reports are available related to combining the nutritional aspects of milk, finger millet and fermentation. Finger millet (Eleusine Coracana) enriched fermented milk product prepared using an indigenous probiotic bacteria could be a novel concept of a functional food. Being a rich source of calcium and iron, and the fact that the bioavailability can be improved by simple processing such as germination and fermentation, the resultant product can be a good supplement for improving bone health and hemoglobin. Hence, the present study was planned and executed to develop a finger millet enriched probiotic fermented milk product.ThesisItem Open Access DEVELOPMENT OF GREEK YOGHURT TYPE PROBIOTIC FERMENTED MILK USING INDIGENOUS CULTURES(DEPARTMENT OF DAIRY MICROBIOLOGY SMC COLLEGE OF DAIRY SCIENCE ANAND AGRICULTURAL UNIVERSITY ANAND, 2017) DESAI RACHANA RAMESHCHANDRA; Dr. Sreeja V.Rising awareness about the health benefits of fermented milk products, increasing disposable income levels, growing demand from middle class segment for health benefitting foods with affordable prices are some of the factors driving demand for fermented milk market in india. According to a report by TechSci Research (2016), India’s yoghurt market is projected to grow at faster pace over next five years and touch US$1 billion by 2021. Hence the time is apt for introducing new varieties of value added fermented milk products to the Indian food market. Current decades saw an increase in the popularity of concentrated yoghurts such as Greek yoghurt which has a number of proposed health benefits such as low lactose and low sodium content, high satiety index as well as the benefits of fermentation. With the exception of shrikhand no other fermented concentrated milk products are available in India. Chakka, the intermediate product obtained by partial removal of whey from curd during shrikhand manufacturing has not been explored as a main product till today. Hence, looking to the market opportunities, consumers demand for novel products, the nutritional and therapeutic benefits of fermented dairy products as well as lack of research reports on (i) systematic studies done on concentrated fermented milks other than shrikhand and (ii) use of buffalo milk for preparation of concentrated fermented milks, the present study was planned and executed to develop a probiotic Greek yoghurt type product from buffalo milk using indigenous starter cultures.ThesisItem Open Access STUDIES ON ENHANCING THE SHELFLIFE OF FERMENTED MILKS(AAU, Anand, 2004) BEHARE, PRADIP V.; Prajapati, J. B.Poor shelf-life of fermented milk hamper their commercial exploitation. Many ways are available to improve shelf-life of fermented milks, but they require compromise with either product quality or starter bacteria. The present investigation was planned to standardize the process for buttermilk manufacture and subsequently, enhance its shelf life by heat treatment, with possibility of maintaining live beneficial bacteria in the product. Initially, incorporation of three different levels of mozzarella cheese whey at 10, 20 and 30 % were tried. Addition of whey at 20 % level in to milk did not affect flavour, body and texture of the curd. A combination of Str. thermophilus MD2, Str. thermophilus Di6 and Lactobacillus acidophilus V3 was used to ferment milk. To select the heat treatments, survivability of the cultures (MD2, Di6 and V3) against different heat treatments were tried. The treatment 55°C for 5 min and 60°C for 5 min left sizable viable population, while 70°C for 5 min was drastic for the cultures. The shelf-life of control (Ti) and treated [55°C for 5 min (T2), 60°C for 5 min (T3) and 65°C for 5 min (T4)] buttermilk was studied at room (37±2°C) and refrigeration temperature (7±2°C) on the basis of sensory, chemical and microbial changes. In sensory changes, flavour was the deciding factor for shelf-life and decreased during the storage gradually at 7±2°C and rapidly at 37±2°C. Body, texture, colour and appearance scores were not affected during storage. Based on sensory attributes, Ti and T2 remained acceptable for 1 day while T3 and T4 were acceptable for 2 days at room temperature. In case of refrigerated storage, Ti and T2 remained acceptable for 21 and 28 days, respectively whereas T3 and T4 remained acceptable even on 35th day. Titratable acidity increased gradually during storage at 7±2°C and sharply at 37±2°C. The similar but opposite trend was seen in pH. FFA and soluble nitrogen increased at faster rate at 37±2°C and steadily at 7±2°C. However, they did not correlate with other changes. Total lactic count of fresh buttermilk was 65.17 x 107 cfu/ml, which reduced to 33.3 X lO7 cfu/ml by T3 and significantly reduced to 27.4 x 107 cfu/ml by T4. At room temperature storage lactic count significantly increased, while at 7±2°C it remained almost stable. The lactobacilli count during the storage at 37±2°C increased sharply, whereas, it remained unchanged at 7±2°C. The survival rate in T3 was 35 % whereas 99.7 % destruction was given by T4. Yeast and mold significantly reduced by T3 and T4. However, T2 was not sufficient to kill yeast and mold. This group was the main culprit in spoiling the product. Results of pilot scale study in a commercial dairy plant gave a shelf life of 15 days for control and 25 days to buttermilk heat treated at 60°C for 5 min.ThesisItem Open Access METAGENOMIC ANALYSES OF GERIATRIC GUT MICROBIOME DURING PROBIOTIC Lactobacillus helveticus MTCC 5463 INTERVENTION(AAU, Anand, 2014) SENAN, SUJA; Prajapati, J. B.Age related changes in the gastrointestinal tract, as well as changes in diet and host immune system reactivity, inevitably affect gut microbial population composition. Therapeutic strategies to counteract these changes have been suggested in ageing people. These include dietary supplements contairung prebiotics, probiotics and a combination of both of these, synbiotics. In this study a double blind randomized crossover trial was carried out where 72 elderly subjects were fed with fermented drink containing probiotic Lactobacillus lielveticus MTCC 5463 for 30 days. A real-time quantitative PCR assay based on bile salt hydrolase gene targeting primers and 3' minor groove binder (MGB) probes for accurate detection and quantification of Lactobacillus helveticus MTCC 5463 in human faecal samples was developed. Out of the 57 strains of Lactobacilli tested by in silico PCR, only two strains L. helveticus HIO and L. helveticus R0052 showed the predictive amplification while the rest 55 tested negative. The primers do not amplify any strains of Streptococcus thermophilus that were added to the fermented milks as starter culture. Genomic DNA standards were prepared with six different serial dilutions (2.68 X 106 to 2.68 X 10). The curve was found to be linear, with R2 values > 0.98, over the ranges of 10^ to lOi CPU/ml for MTCC 5463 strain. The slope of the standard curve was -3.372 which predicted the assay efficiency as 97.95%. At the end of 30 days the strain appeared in the faeces of all subjects in the treated group, reaching a level as high as 8.32 to the lowest amount of 6.17 log gene copies/g faecal matter at end of feeding period. After wash out, L. lielveticus MTCC 5463 was detected at gradually reduced levels. It can be observed that the lowest count of probiotic strain post washout was 3.75 log gene copies/g faecal matter at the end of 8 weeks of discontinuation of feeding. This proves the trarisient and consumption-dependent nature of this probiotic bacterium. The strain was not detected in any of the subjects before active test feeding. Traditional plate counts of lactobacilli at genus level on selective medium ranged from a baseline reading of 8.6 log CFU/g of wet fecal matter, which rose to 9.3 log CFU/g at the end of feeding period and a gradual decrease to 8.7 log CFU/gm at the end of the placebo feeding. The qPCR primers targeted the bile salt hydrolase gene of MTCC 5463 which made the gene copy count a fraction of the plate count. From the plate count results it can be said that the recovery of Lactobacilli from stool samples was at IxlO^ colony forming units/g (CFU/g) by week 4 giving a recovery of 82%. A more precise picture of the recovery of the strain calculated using qPCR results came to 18%. Among the 72 elderly subjects who participated in the trial, we could identify 10 respondents who showed positive results in the primary outcome of cholesterol reduction and 10 who showed an increase in cholesterol with a decreasing lactobacilli population indicating non response to probiotic therapy. DNA from the faecal samples of these 20 respondents during baseline and end of feeding was analyzed. Amplicons from the hypervariable region of the 16S rRNA gene were generated and sequenced each on a 316 chip. The data sets for before feeding have a total of reads ranging from 13,061 to 980,628 with read length ranging from 201 to 251 bps and a total amount of 42,52,62,470 bases. The data sets for after feeding showed reduction in reads ranging from 65 to 102,507 with read lengths varying from 268 to 165 with a total of 59,962,912 bps. After a washout of 4 weeks and before placebo feeding the sequencing data can be summarized having reads ranging from 1.386 to 172,304 with read lengths of 158 to 198 bps and a total of 24,10,52,754 bps. Post placebo feeding saw a similar trend of reads from 425 to 171,896 with sequence length from 133 to 155 bps with a total base pairs of 29,14,22,863. Sequencing reads were clustered into operational taxonomic units described by community metrics and taxonomically classified. Reads per sample were clustered and studied for diversity and richness using MG-RAST. All the community members in our samples were from the domain bacteria. The most prevalent phyla in all samples were: Firmicutes, Proeohacteria, Actinohacteria and Baderoidetes with Firmicutes dominating in all samples. All the samples taken prior to treatment showed an abundance in Blautia, Bifidobacterium, Clostridium, Escherichia, Eubacterium, Faecalibacterium, Lactobacillus, Prevotella, Roseburia, Ruminococcus and Shigella. It was strikingly evident that the non respondents harboured more Shigella, Escherichia and less Runinococcus and Clostridium (compared to positive respondents). Lactobacilli and Prevotella showed an increase in abundance values after probiotic treatment with a decrease in Shigella, Ruminococcus, Bacillus and Bifidobacterium. The shifts in gut community structure during probiotic therapy was also studied using QIIME, an open-source software pipeline able to perform, starting from raw sequence data, a wide range of analyses on microbial communities, that is, sequence aligiunent, identification of operational taxonomic units (OTUs), elaboration of phylogenetic trees, and phylogenetic or taxon-based analysis of diversity within and between samples. The responders showed 52.1 % Firmicutes compared to 40.6 % in non responders. The other major phyla Proteobacteria was higher in non responders at 49% than 38% in responders. The class based assignments of responders showed profound shifts in Bacilli, Clostridia and Gammaproteobacteria. Among non responder subjects, the relative proportion of Lactobacillales, Clostridials and Enterobacteriales could be the deciding biomarkers as in the case with responders. We could assume that interplay of Firmicutes and Proteobacteria and specific classes like Clostridiales, Enterobacteriales and Lactobacillales could be indicative of the amenability of the gut microbiota to dietary modification. To examine the difference in responders and non responders for the genera of major phyla Firmicutes and Proteobacteria we used STAMP software for statistical techiuques. We could identify a few microbial biomarkers that differentiate the responders from non responders. The STAMP analysis revealed that among responders and non responders the chief genera of Firmicutes that showed significant difference are Lactobacillus, Clostridium, Euhacterium, and Blautia (q< 0.002) while the genera of Proteobacteria included Shigella, Escherichia, Burkholderia and Camphylobacter (q-value<0.002). This proof-of-principle study introduces for the first time in India, potential microbial biomarkers for probiotic MTCC 5463 responsiveness in geriatric volunteers and reveals the potential of microbiota signatures for personalized nutrition.ThesisItem Open Access DEVELOPMENT OF PROBIOTIC ICE-CREAM SUPPLEMENTED WITH FINGER MILLET (RAGI)(AAU, Anand, 2013) CHAUDHARY, MILTESHKUMAR GANESHBHAI; Prajapati, J. B.Designing of functional foods with probiotics for health benefits is gaining interest in recent year. Ice-cream is a very popular product and hence, has been evaluated as one of the matrices which could carry probiotic bacteria. In this context a study was taken up to develop a probiotic ice-cream supplemented with finger millets (ragi) by using indigenous probiotic cultures and evaluate for survival of probiotic bacteria and sensory attributes. Constituents of ragi flour were modified with the help of germination of ragi grains followed by grinding and designated as malted ragi flour (MRF). Standardized ice-cream mix was formulated in order to obtain 6.0% Fat, 11.0% MSNF, 4.9% Protein and 15% Sucrose in ice-cream. MRF was added (9% w/w) into the ice-cream mix and gelatinization of ragi flour was carried out by heating part of the mix containing MRF upto 90°C without holding. The ice-cream mixes were aged for 4 ± 2 °C for over night and used to prepare ice-creams. Active strains of probiotic culture Lb. helveticus MTCC 5463 and Lh. rhmnosus MTCC 5462, separately were added at the rate of 0.1% (w/v) of ice-cream mix in the form of concentrated cells (CC) as well as freeze dried powder (FDP) to ice-cream mix soon after the ageing. Survival of probiotic culture was higher {P<0.05) upon freezing of ice-cream mix when added as CC (91-95%) as compared to FDP (89-90%) regardless of types of probiotic strain. Lh. rhamnosus MTCC 5462 was observed to be less resistant to the process of ice-cream manufacturing as compared to Lb. helveticus MTCC 5463, regardless of its form. With regard to sensory attributes, there was no statistical difference among the samples (P > 0.05), indicating that neither the addition of Lh. helveticus MTCC 5463 nor Lh. rhamnosus MTCC 5462 nor the forms of probiotic strains influenced the overall acceptance of ice-cream as compared to control sample. Based on higher survival, probiotic culture Lb. helveticus MTCC 5463 was selected to manufacture ice creams with or without ragi. These ice-cream samples were compared with non-probiotic ice-cream in order to evaluate their sensory attributes, pH and survival of Lb. helveticus MTCC 5463 upon frozen storage of 90 days. It was observed that addition of Lb. helveticus MTCC 5463 (CC) did not significantly (P<0.05) affect the pH of the ice-cream up to 90 days storage. Viability of Lb. helveticus MTCC 5463 was higher in the ice-cream supplemented with ragi (96%) as compared to plain ice-cream (88%), suggesting protective effect of ragi principles on lactobacilli. The counts of lactobacilli significantly (P < 0.05) decreased in the range of 0.38-0.65 and 1.1-1.54 log cfu/g in probiotic ragi ice-cream and plain probiotic ice-cream respectively at the end of 90 days storage. However, freezing and mixing involved in converting the mix into ice-cream was found to impart a greater (P < 0.05) effect on culture viability, than storage in ice cream for both the ice-cream. Viable counts of lactobaciUi remained above 7 log cfu/g for both plain and ragi ice-cream at the end of storage period. The changes in the flavour score amongst all treatments did not differ significantly (P> 0.05) over the period of 90 days. Average score for colour and appearance of probiotic and non probiotic ice-cream (with or without ragi) decreased significantly (P< 0.05) over the period of time. Mean score of body and texture for probiotic and non probiotic ice-cream (with or without ragi) not differed significantly (P> 0.05) up to 60 days. However, the difference was significant after 90 days. Average of melting quality score for probiotic and non probiotic ice-creams (with or without ragi) were above 7 which showed high melting quality of all the ice-creams through the storage. Overall acceptability for probiotic and non probiotic ice-cream (without ragi) ranged from 7.86 and 7.81 respectively, whereas the score for probiotic and non probiotic ice-cream (supplemented with ragi) was in the range of 8.19 and 8.21 respectively which decreased significantly (P< 0.05) over the period of 90 days storage. Less than 10 coliforms per g were detected in fresh sample of ice-cream, which disappeared after 30 days of storage. Yeast and molds were absent in 1 g of all the ice-cream samples during storage period. It could be concluded that probiotic ice-cream containing Lb. helveticus MTCC 5463 and supplemented with 9% (w/w) ragi coiild be prepared by incorporation of concentrated cells of the culture at the rate of 0.1% (w/v) of the ice-cream mix just before the freezing. The ice-cream was acceptable till 90 day of storage at -18°C and the viability of probiotic culture remained above 8 log cfu/g, that is higher than the therapeutic minimum throughout the storage.ThesisItem Open Access STUDIES ON ENHANCING THE SHELFLIFE OF FERMENTED MILKS(AAU, Anand, 2004) BEHARE, PRADIP V.; Prajapati, J. B.Poor shelf-life of fermented milk hamper their commercial exploitation. Many ways are available to improve shelf-life of fermented milks, but they require compromise with either product quality or starter bacteria. The present investigation was planned to standardize the process for buttermilk manufacture and subsequently, enhance its shelflife by heat treatment, with possibility of maintaining live beneficial bacteria in the product. Initially, incorporation of three different levels of mozzarella cheese whey at 10, 20 and 30 % were tried. Addition of whey at 20 % level in to milk did not affect flavour, body and texture of the curd. A combination of Sir. thermophilus MD2, Str. thermophilus Die and Lactobacillus acidophilus V3 was used to ferment milk. To select the heat treatments, survivability of the cultures (MD2, Die and V3) against different heat treatments were tried. The treatment 55°C for 5 min and 60°C for 5 min left sizable viable population, while 70°C for 5 min was drastic for the cultures. The shelf-life of control (T1) and treated [55°C for 5 min (T2), 60°C for 5 min (T3) and 65°C for 5 min (T4)] buttermilk was studied at room (37±2°C) and refrigeration temperature (7±2°C) on the basis of sensory, chemical and microbial changes. In sensory changes, flavour was the deciding factor for shelf-life and decreased during the storage gradually at 7±2°C and rapidly at 37±2°C. Body, texture, colour and appearance scores were not affected during storage. Based on sensory attributes, T1 and T2 remained acceptable for 1 day while T3 and T4 were acceptable for 2 days at room temperature. In case of refrigerated storage, T1 and T2 remained acceptable for 21 and 28 days, respectively whereas T3 and T4 remained acceptable even on 35th day. Titratable acidity increased gradually during storage at 7±2°C and sharply at 37±2°C. The similar but opposite trend was seen in pH. FFA and soluble nitrogen increased at faster rate at 37+2°C and steadily at 7±2°C. However, they did not correlate with other changes. Total lactic count of fresh buttermilk was 65.17 x 10 to power 7 cfu/ml, which reduced to 33.3 X 107 cfu/ml by T3 and significantly reduced to 27.4 x 10 to power 7 cfu/ml by T4. At room temperature storage lactic count significantly increased, while at 7±2°C it remained almost stable. The lactobacilli count during the storage at 37±2°C increased sharply, whereas, it remained unchanged at 7±2°C. The survival rate in T3 was 35 % whereas 99.7 % destruction was given by T4. Yeast and mold significantly reduced by T3 and T4. However, T2 was not sufficient to kill yeast and mold. This group was the main culprit in spoiling the product. Results of pilot scale study in a commercial dairy plant gave a shelf life of 15 days for control and 25 days to buttermilk heat treated at 60°C for 5 min.