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ThesisItem Open Access INCLUSION OF TASTE AND AROMA IN THE TOTAL MIXED RATION AND THEIR EFFECT ON DIETARY PREFERENCE AND GROWTH PERFORMANCE OF CALVES(ICAR-NDRI, KARNAL, 2023) NALIYAPARA HARDIKKUMAR BHUPATBHAI; RAMAN MALIKThe present investigation was conducted to evaluate the inclusion of taste and aroma in the total mixed ration (TMR) and their effects on dietary preference and growth performance of calves. The study was divided into two phases. In Phase 1, three different levels of five tastes and five artificial feed aromas were added in paddy straw-based TMR’s and their effects on dietary preference and feed intake of growing KF calves were studied using a sequential elimination trial. Tastes include sweet (sucrose), umami (MSG), salty (NaCl), bitter (urea) and sour (citric acid). Whereas aromas include lacto-vanilla, maize, guar korma, green grass and silage. At inclusion level 1, TMR was mixed with i. Sucrose, ii. MSG, iii. Sodium chloride, iv. Urea, and v. Citric acid at rates of 50, 10, 15, 5 and 10 g/kg DM, respectively. However, rate of artificial feed aromas inclusion was 2.5 g/kg DM. Ingredient dose rates for tastes and feed aroma were further increased by 25% and 50%, respectively, at inclusion levels 2 and 3. All taste ingredients and feed aromas were dissolved in water and sprayed on paddy straw-based TMR (the ratio of roughage to concentrate was 70:30). For the animal trial, six healthy growing KF calves were selected and each kept in an individual pen. The sequential elimination trial was conducted in five slots. The duration of slots 1,2,3,4 and 5 was six, five, four, three, and two days, respectively. During the sequential elimination trial on tastes, six paddy straw-based TMR’s were offered to each calf, in which five TMR had a different taste and one was without taste. After completing slot 1 of six days, the most preferred diet with the highest DMI was eliminated. Likewise, the remaining five feed was fed for five days in 2nd slot, four feed was fed for four days in 3rd slot, three feed was fed for three days in 4th slot and two feed was fed for two days in 5th slot. The same trial was conducted for three different levels of inclusions of aroma. It was found that among the taste, sweet was the most preferred (p<0.01) taste, followed by umami, salty, bitter, sour and control. While, in inclusion level 3, DMI (kg/100kg BW; g/kg BW0.75) was decreased (p<0.01) and TMR without taste was more preferred than sour and salty in the 4th and 5th slot. Among the feed aromas, preference ranking, lacto-vanilla and maize were comparable, followed by guar korma, green grass, silage and control. In contrast, the level of aroma inclusion did not affect the DMI (kg/100kg BW; g/kg BW0.75). In Phase 2, a growth trial on 24 healthy KF calves was conducted to know the effects of the inclusion of sucrose as a sweet taste and lacto-vanilla aroma on feed intake, digestibility, blood biochemical parameters, and growth performance of animals. The selected animals were divided into four treatment groups based on their body weight. Each group had six animals. T1 was kept as a control, T2 was fed TMR having additional 62.5 g sucrose per kg DM, T3 was fed TMR by adding 3.125 g lacto-vanilla aroma per kg DM, and T4 was fed TMR with the combination of both sucrose and lactovanilla. The study revealed that net weight gain and average daily weight gain were higher (p<0.001) in the taste and aroma-supplemented groups. Significantly higher dry matter intake (kg/100kg BW; g/kg BW0.75) was observed in T2 and T4 groups, and DMI in T3 was at par with other groups. At the same time, digestibility and nitrogen balance were not affected by the taste and aroma supplementation. Significantly higher (p<0.01) ghrelin concentration in blood plasma was observed at 0 hr before and 4 hr after feeding, whereas leptin concentration was increased (p<0.01) 2 hours after feeding. Glucose, total protein, albumin and globulin level did not differ significantly among the groups. Creatinine, BUN, AST and ALT level were found within the normal range and also did not differ significantly among the groups.ThesisItem Open Access EFFECT OF SILICON SUPPLEMENTATION ON THE MINERAL METABOLISM AND GROWTH PERFORMANCE IN CALVES(ICAR-NDRI, KARNAL, 2023) LAMELLA OJHA; Raman MalikThe present study was carried out to see the effect of silicon supplementation on the mineral metabolism and growth performance in calves. The study was conducted in two phases. In the phase first, determined the silicon (Si) content of various feeds and fodders and analyzed their bioaccessibility (%) and also estimated the solubility (%) of 4 different Si containing salts at different pH. Results showed that dry as well as green fodders contained more silicon than concentrate feedstuffs, however, the highest percentage of bioaccessibility was seen in grains and grain by-products followed by unconventional feedstuffs and forages. The mean solubility was significantly higher for orthosilicic acid and the lower values were observed for sodium zeolite A while metasilicate and tetraethyl orthosilicate are intermediate. In 2nd phase two animal trials (pre ruminant and ruminant) were conducted in Karan Fries calves. In 1st animal trial, 24 calves (5-7 day) were selected on the basis of their BW and divided into 4 groups (T1, T2, T3 and T4) and fed as per ICAR (2013) feeding standards except that these were additionally supplemented with 0, 50, 100 and 150 ppm (as orthosilicic acid) in four respective groups for 90 days. Results revealed that growth performance, DMI were significantly increased in T2, T3 and T4 groups, whereas DMI (% BW), nutrient utilization, digestibility coefficient, N and minerals balance (P, Mg, Zn,Cu, Mn and Fe), haemato-biochemical parameters, immune response and antioxidant profiles were not altered due to Si supplementation. Silicon increased the balance as well as plasma levels of Si, Ca and B, concentration of plasma growth hormone, vitamin D3, bone alkaline phosphatase and osteocalcin in T3 and T4 groups, but the levels of calcitonin, PTH and hydroxyproline were remain similar among all the groups. Thus, the inclusion of 100 and 150 ppm of Si was effective in improving the growth performance, mineral balance (Ca and B), growth hormone, vitamin D3 and bone health status in pre ruminant calves. In trial second, 24 calves (3.5 months) were randomly distributed in 4 groups (T1, T2, T3 and T4) and fed basal diet as per ICAR (2013) feeding standards. Silicon was additionally supplemented with 50, 100 and 150 ppm in T2, T3 and T4 groups for 120 days. Results revealed that BW, DMI, nutrient utilization, digestibility coefficient, N and minerals balance (P, Mg, Zn,Cu, Mn and Fe), haematobiochemical parameters, immunity, antioxidant status and hormonal profile were not influenced due to Si supplementation. However, Si increased the balance and plasma Si, Ca and B concentration, plasma levels of bone biomarkers (BALP and osteocalcin) in T3 and T4 groups, but hydroxyproline levels was not altered among all the groups. The results indicated that supplementation of Si at 100 and 150 ppm had increase the mineral balance (Ca and B) and bone biomarkers in ruminant calves. However, supplementation of 150 ppm had no additional benefit, therefore 100 ppm is the optimum level of Si supplementation, particularly in both pre-ruminant and ruminant calves.ThesisItem Open Access NUTRITIONAL STATUS OF FARM WOMEN IN ASPIRATIONAL DISTRICTS OF KERALA AND TAMIL NADU(ICAR-NDRI, KARNAL, 2023) DITTY MARIA DOMINIC; H. R. MeenaAbout two third of the labour in farming is constituted by women. Irrespective of their degree of affluence, they provide 14 to 18 hours of productive physical labour every day. Nutrition has notifying significance in case of farm women, because they have to bear and rear children along with carrying out agricultural production activities. Hence the present study focused on the nutritional status of farm women in aspirational districts of Kerala and Tamil Nadu. The study was conducted in Wayanad, Virudhunagar and Ramanathapuram districts, which were listed as aspirational districts by NITI Aayog. From each aspirational district, two blocks were randomly selected. Two villages were selected from each block randomly and 30 households having at least 2 milch animals and cultivating in at least one crop season were selected from each village as respondents. Thus the total sample size was 360 households. Nutritional status of farm women was measured using Body Mass Index (BMI) (anthropometric measure). Household food security was measured using Household Food Insecurity Access Scale (USAID, 2007), the factors affecting nutritional status was determined using exploratory factor analysis followed by multinomial regression. Behavioral intention to consume nutritious food was studied using Likert type scale based on theory of planned behavior (Ajzen, 1985) and structural equation modeling was used for theory testing. A multi-lingual education module (folder) was developed on diet and nutrition of farm women and its perceived effectiveness was measured using the scale developed based on Guidelines for Pretesting and Evaluating Communication Materials given by Ministry of Health, Zambia and USAID, as it is cross country validated and reliability was high. Major findings of the study were as follows: Majority of the households (~67%) experienced various levels of food insecurity (moderate [35.83%] –mild [25.83%]-extreme level [5.28%]) and only 33 percent of the households were food secure. Majority of the respondents (59-95%) did not experience severe food insecurity. There is a significant but weak relationship between food security and nutritional status. All the three identified factors had significant influence in the nutritional status of farm women in both the states and cumulatively explained the variance in BMI up to 59 percentage. In both the states majority of the respondents had medium intention to consume nutritious food, followed by high intention. The model fit indicators shows that theory of planned behavior adequately predicts behavioral intention. The effectiveness of the module was perceived as high by the respondents in general. Hence the study reinforces the need for nutrition sensitive extension approaches.ThesisItem Open Access FARM DIVERSIFICATION IN HARYANA: AN APPRAISAL FOR FUTURE FARMING(ICAR-NDRI, KARNAL, 2023) ELIZABETH JOSE; K. PonnusamyContinuous mono cropping system threatens the sustainability of future agricultural production in terms of soil, water, climate and market parameters which warrants diversified farming system enabling farmers to use their resources efficiently with its ecological benefits. The present study was undertaken to explore the potential of farm diversification in the state of Haryana. The study was carried out during 2020-2022 in three agro ecological zones of Haryana state. One district from each agro-ecological zone was chosen followed by two blocks each in every district and two villages of selected block. With the help of expert opinion and progressive farmers, 15 farmers from each village were selected. Ex-post design and case study approach were followed. The data were collected using structured interview schedule with the sample size of 240 comprising 180 farmers and 60 experts. Data were analysed by frequency, percentage and Cumulative Square Root Technique. Two Delphi surveys were organized with 60 experts to identify future drivers of Haryana agriculture sector for the year 2030. The study revealed that Crop Diversification Index of the Haryana state was found to be decreasing (0.272(2010) to 0.262(2018)), while Farm Diversification Index of the state was found to be increasing (0.509(2010) to 0.546(2019)). Proximity to NCR (0.896), increased road density (0.857) and improved access to market information (0.801) were found to influence farm diversification extensively. Market uncertainties (0.876) and climate induced factors (0.796) were perceived to hinder the farm diversification considerably. Farm diversification with enterprises such as sheep (3-3.8), piggery (2.4-2.66), apiary (2.55), mushroom (4.8), ber (5-6) and cauliflower (2.6-5) provided better income in comparison to the traditional field crops. Kaithal district (2.46) reported the highest farm diversification efficiency followed by Bhiwani (1.88) and Hisar (1.64) districts. Availability of fertile soil, increased area under irrigation, better marketing and transportation facility were the major reasons for Kaithal to score the highest BC ratio compared to Hisar and Bhiwani. The study suggests the need for modification in government schemes (MPMV and BBY), making provision for adequate attractive incentives and timely payment of the compensation amount. Extension efforts should be fine-tuned for accelerating the adoption of enterprises such as poultry, piggery, fishery, apiary, mushroom, ber and cauliflower and sheep farming due to their potential for higher income generation and suitability for farm diversification. R and D efforts should be strengthened to enhance the income from dairy farming. Scenario forecasting on future farming of Haryana for the year 2030 reveals that if government policy support and technological up-gradation moves in an accelerated way (Shining future), there is a high chance of large scale production of food and non-food crops, milk, egg and meat. In addition, government policies are also focusing on the production of less water requiring crops, which in turn decelerate growth of paddy in this scenario. Study concludes that area based planning (based on SWOT analysis of different agro-ecological zones of Haryana) is the prime strategy to achieve farm diversification in the state. Farmer Producer Companies, Public Private Partnership, ICT tools, entrepreneurship and advanced farm machines should be the areas of attention for developing farming sector of Haryana state in a sustainable fashion.ThesisItem Open Access EXPLORATORY ANALYSIS OF INTEGRATED FARMING SYSTEMS OF KERALA(ICAR-NDRI, KARNAL, 2023) VANI CHANDRAN; Ritu ChakravartyThe present study was conducted in Kerala. Three districts were selected randomly each from Southern, Central and Northern Kerala. The respondent groups of the study were comprised of Farmers and Extension Personnel. The total sample size was 240, among that 180 were IFS farmers and 60 were officials. Data were collected through a well structured interview schedule. The collected data were analyzed by using appropriate statistical tools in order to draw meaningful conclusions. Most of the IFS farmers in Kerala belonged to old aged category and had completed education up to higher secondary. The primary occupation of the majority of the respondents were farming only and they were marginal farmers with small family size and herd size. Most of the farmers had medium level experience in IFS activities and had high mass media exposure with medium extension agency contact and extension participation. In the study area, nine IFS components were identified. Component analysis in terms of extent of adoption, integration of practices and contribution to household income revealed that, extent of adoption of identified components and contribution to annual household income varied from low to medium whereas extent of integration of available components varied from medium to low. Based on various combinations, eight different dairy based IFS systems were identified and dairy and crop were the dominant component in all systems both in terms of adoption as well as integration. The findings also pointed out that for establishing and maintaining various components in a unit, farmers were experiencing different technological needs and constraints. IFS farmers perceived the need for value addition technologies more than production technologies. In general, they preferred socially accepted low cost technologies with high income generation capacity, efficiency, flexibility, easy accessibility and permitting sustainable and maximum utilization of local resources. Lack of remunerative prices for farm produces and high cost of production were the top ranked constraints experienced by the farmers. Majority of the IFS farmers had a neutral to favourable attitude towards IFS. IFS offers multiple sources of income and guarantees supply of balanced and nutritious food to families which were the most important factors that influenced the attitude. The findings highlight the need for revamping of the existing policies, schemes, programs, to support institutional strengthening and capacity building of the farmers. This will help the farmers to expand their existing units by adding more components and technologies to their units. Further, this would overcome many constraints faced, thereby improving the productivity and profitability.ThesisItem Open Access INDIGENOUS CATTLE-REARING AMONG THE DAIRY FARMERS: AN EXPLORATORY STUDY IN KARNATAKA(ICAR-NDRI, KARNAL, 2023) CHETHAN PATIL N. D.; K.S. KADIANThe present study entitled Indigenous cattle-rearing among the dairy farmers: An exploratory study in Karnataka was taken up with the aim of exploring livelihood of indigenous cattle keeping farmers, document existing management practices in rearing cattle, production performance, preferential traits of breeds, attitude towards rearing indigenous cattle, constraints in cattle-rearing and efforts of different agencies in conservation of indigenous cattle. Exploratory and ex-post facto research design with case study method was employed. The study was purposively conducted in six districts of Karnataka which is having three registered indigenous cattle breeds namely, Amritmahal, Hallikar and Malnad Gidda. One hundred twenty indigenous cattle owners from each breed were selected thereby making a total of 360 respondents. Data were collected using semistructured interview schedule by personal interview method. Respondents with a mean age of 42.96±0.45 years with primary to secondary school level education, nuclear family type with small to medium family size, having annual income of medium to the low, land holding of semi-medium to small, mass media exposure of very poor to poor and low to medium social participation, medium to low extension agency contact, low to medium level of risk orientation, innovative proneness and economic motivation. From indigenous cattle, each household earned an average of ₹16876 per annum, with a mean share of 24.82% in annual family income. Annually, around 198 man-days of employment were generated from indigenous cattle. The average daily consumption of indigenous cow milk was 1.22 liters/household. The annual mean FYM production was 5.98 tons per household. Around 56.67% of the respondents considered rearing indigenous cattle as the security against uncertainties and 63.33% symbolized social status. High adaptability was first ranked preferred trait in indigenous cattle followed by multi-utility, easy maintenance, traction power and religious/cultural sentiments. Significant differences were found between the breeds with respect to age at first calving, lactation length, dry period, inter-calving period and service period. Majority of the respondents using both natural service and artificial insemination in indigenous cattle breeding. Grazing cum stall feeding was commonly practiced feeding method in all the breeds except in Malnad Gidda, wherein 95% depend on open grazing. Most of the respondents feed concentrates and green fodder to indigenous cattle on seasonal/ production basis. Mixed type of cattle shed was commonly seen, located adjacent to farmers’ residence. Vaccination of indigenous cattle was widely practiced with occasional deworming. Cent per cent of respondents feed colostrum to new-born but only after expulsion of placenta. Knuckling and stripping were common milking methods being followed. Producer-Brokers-Buyers link was the common marketing channel for trade of indigenous cattle under unorganized local livestock market. Four separate case studies were also done including a Math, a goshala, a government cattle breeding farm and a progressive farmer, all of whom are working for conservation and development of indigenous breeds of Karnataka. The study concluded that indigenous cattle keeping happened to be one of the important livelihood sources for the farmers. Along with supplementing family income, they also play a major role in achieving the nutritional security, enriching land through FYM, providing employment opportunity around the year for better utilization of family labour and act as security during uncertainties. Majority of the respondents had neutral attitude towards rearing indigenous cattle and pluralistic agencies like SDAH, religious institutes, NGO and progressive farmers working for conservation of indigenous cattle; but it needs extension interventions for convergence of their efforts.ThesisItem Open Access ATTENUATED TOTAL REFLECTANCE-FOURIER TRANSFORM INFRARED SPECTROSCOPY COUPLED WITH CHEMOMETRICS TO DETECT SELECTED ANIMAL BODY FATS, VEGETABLE OILS AND THEIR ADMIXTURE IN GHEE(ICAR-NDRI, KARNAL, 2022) VIVEK SONVANSHI; KAMAL GANDHIGhee is a popular traditional dairy product of India. Due to its high demand and insufficient supply during lean season, it becomes prone to adulteration by unscrupulous traders in the market. The present study was conducted to develop suitable models using ATR-FTIR coupled with chemometrics for detection of selected animal body fats, vegetable oils and their admixture in ghee. Milk samples were procured and ghee samples were prepared using creamery butter method. Vegetable oils from five reputable brands and animal body adipose tissues were purchased from local market of Karnal. Animal body fats were extracted from the respective adipose tissues of the animal using dry rendering process. GC-analysis revealed that the short chain fatty acids were only present in pure ghee and not in adulterants targeted in the study. Linoleic acid concentration was significantly higher in soybean oil as compared to that in other oils and fat studied. Palmitic acid was the major fatty acid in vanaspati and trans fatty acids (eladic acid) was only present in vanaspati. Goat body fat contained higher amount of stearic acid as compared to that in other oils/ fats studied. Oleic acid concentration was higher in palmolein oil and pig body fat as compared to that in other oils and fats studied. Prepared cow and buffalo ghee were mixed in equal proportions to obtained pure mixed ghee (PMG). Adulterants, viz. soybean oil (SO), vanaspati (VG), palmolein oil (PO), goat body fat (GBF), pig body fat (PBF), sheep body fat (SBF) were added individually at 1, 3, 5, 10, 15 and 20 as well as in combinations, viz. SO+GBF, VG+GBF, PO+GBF, SO+PBF, VG+PBF, PO+PBF, SO+SBF, VG+SBF and PO+SBF in PMG in the ratios of 1:2.3, 2:4.6, 3:7, 4:9.3 and 5:11.6, respectively. FTIR spectra analysis of all the samples were performed and the data obtained was subjected to chemometric analysis. Functional group regions of the mixed ghee and adulterants were almost similar while slight differences in their finger print regions were observed. Wavenumber regions which were found useful for detecting adulteration of ghee with SO, VG, PO, GBF, PBF and SBF were 727-702, 1120-1080 and 985-955, 1167-1137, 1760-1730, 1190-1140 and 1100-970, 1190-1140 and 1120-970 and 732-710 cm-1, respectively. Wavenumber regions which were found useful for detecting adulteration of ghee with admixture of SO+GBF, VG+GBF, PO+GBF, SO+PBF, VG+PBF, PO+PBF, SO+SBF, VG+SBF and PO+SBF were 1180-1140 and 1120-1098, 1170-1145 and 1120-1087, 1175-1135 and 1125-1080, 1170-1140 and 1130-1090, 1180-1140 and 1120-1090, 1200-1130 and 1123-1093, 730-710, 1125-1085 and 740-700 cm-1, respectively. PCA applied in the selected regions showed separate clusters from PMG even for the lowest level of spiking of each adulterant and their admixture and as the level of spiking of adulterants increased, clusters shifted towards pure adulterants. PLS and PCR models applied in the selected regions of the FTIR spectra were equally efficient in detecting the selected adulterants in ghee as indicated from the R2, RMSEC, RMSEV and Bias values. Calibration curves between the actual and predicted levels of all the individual adulterants and their admixture in ghee were linear with a slope of 45° and no x or y intercepts indicating the suitability of the models in detecting them in ghee. SIMCA approach in conjunction with the established PLS models applied in the selected wavenumber regions showed the classification efficiency for pure mixed ghee, pure body fats and pure vegetable oils as 100% indicating that models were effectively developed for their detection in ghee. Classification efficiencies for ghee samples containing individual adulterants and their admixture at all the levels studied were never fell down below 85 and 73%, respectively. Using ATR-FTIR, we can detect up to 1% level of selected individual adulterants and 3.3% level of admixture of animal body fat and vegetable oil in ghee. Rapid, non-destructive, low cost and accurate analytical protocol involving a combined use of FTIR spectroscopy and chemometrics for rapid and accurate determination of soybean oil, vanaspati, palmolein oil, goat, pig, sheep body fat and admixture of SO+GBF, VG+GBF, PO+GBF, SO+PBF, VG+PBF, PO+PBF, SO+SBF, VG+SBF and PO+SBF in ghee is now available which the industries can adopt for regular testing of their samples.ThesisItem Open Access PHENOTYPIC AND GENOTYPIC CHARACTERISATION OF ANTIMICROBIAL RESISTANCE IN ENTEROCOCCUS SPECIES OF DAIRY ORIGIN(ICAR-NDRI, KARNAL, 2022) RAVIKANT VINAYAKRAO VINCHURKAR; DIWAS PRADHANPrevalence of antibiotic resistance among food-based enterococci is matter of concern because of its opportunistic pathogenicity as well as its ability to disseminate AMR genes to other bacteria in the food chain. The present study was undertaken to characterize antimicrobial resistance in Enterococcus species of dairy origin. A total of 64 dairy samples comprising of traditional Dahi and raw milk samples were collected from the districts of Karnal, Kaithal, Sirsa, Bhiwani and Jind of Haryana region. The sample were processed for isolation of enterococci by pour plate technique using selective media viz. Citrate Azide and Bile Esculin Agar and further identified by phenotypic and genotypic approach. A total of 235 isolates were initially shortlisted based on Gram’s staining and catalase test. The genotypic identification of isolates was done by using genus specific PCR. Finally, a total of 140 isolates were genotypically confirmed as enterococci. A total of 84 Enterococcus isolates were further tested for antimicrobial susceptibility against major antibiotic classes as per CLSI guidelines. The highest resistance in the enterococci isolates were observed against Cefuroxime (50%), Rifampicin (21.43%), Cefepime (19.05%), Erythromycin (15.48%), Fosfomycin (9.52%), Cefotaxime (8.33%), as well certain isolates showing resistance against Tetracycline, High Level Streptomycin, Gentamycin, Norfloxacin, Chloramphenicol and Levofloxacin. Extended Spectrum β Lactamases production was confirmed in 9 out of 24 isolates. PCR for antibiotic resistance genes in respective resistant isolates showed positive results for Aminoglycoside resistance genes such as aac(6’)-Ie- aph(2”)-Ia, aph(3’)-IIIa, ant(6)-Ia, Tetracyclines resistance genes [tet(M), tet(O)], Chloramphenicol resistance genes (catA8) and also certain isolates with Macrolide resistance genes such as ermA, ermB, ermC, ermF and msrA. Among the 43 resistant isolates, 69.76% of isolates were positive for multi-drug transporter gene (emeA), while 25.58% and 30.23% were positive for Transposons family, Tn-5397 (tndX) and Tn-916/Tn-1545 (Int-Tn), respectively. Further, 4 isolates also showed the presence of class-1 Integron gene, while 12 isolates were positive for class-3 Integron gene. A total of 43 resistance isolates were tested for virulence traits. Among these only 2 (4.6%) isolates demonstrated Gelatinase production (also positive for gelE gene), 17 (39.53%) for Hemolysin production (6 showed positive for cylA gene) and none for DNase production. No significant correlation between the virulence and antibiotic resistance traits were observed in the isolates. Further, the resistant isolates have been sent for 16S rRNA partial sequencing for species identification and the most resistant isolate B1(C) for Whole Genome Sequencing to study the organization of AMR genes in the isolate.ThesisItem Open Access EVALUATING THE IRON BIOAVAILABILITY FROM EXOPOLYSACCHARIDE KAR1- IRON COMPLEX AND ITS EFFECT ON ANAEMIC RAT MODEL(ICAR-NDRI, KARNAL, 2022) SHASHANK GOWDA B G; PRADIP V. BEHAREIron deficiency anaemia is the most common micronutrient deficiency disorder among people of different age groups, for which effective nutritional strategies are warranted to tackle its devastating effects on public health. The present study investigated the iron bioavailability from an EPSKar1-iron complex using Caco-2 cells and anaemic rats. Exopolysaccharide (EPSKar1) extracted from Lacticaseibacillus rhamnosus Kar1 had the carbohydrate and protein content of 78.33±0.86% and 3.58±0.27%, respectively. Upon purification by ion-exchange chromatography, the carbohydrate and protein concentrations were found to be 95.42±0.04% and 0.30±0.02%, respectively. The purified EPSkar1 was complexed with FeSO4 iron salt and the obtained 100 mg EPSKar1-iron complex contained 53.65±0.04 mg EPSKar1 and 44.26±0.14 mg iron. During simulated gastro-intestinal digestion of 100 mg EPSKar1-iron complex; 9.20±0.23 mg of free iron was released at the intestinal phase corresponding to 20.92±0.53% iron bio-accessibility. When the cytotoxicity of iron and EPSKar1 was evaluated on Caco-2 cells, no significant difference (p <0.05) in the loss of viability was found at the highest concentration of EPSKar1 (75 mg/mL) and FeSO4 (10 mg/mL). In iron bioavailability study using Caco-2 cells, there was a significant increase (p <0.05) in the iron uptake from the cells treated with EPSKar1-iron complex (61.27±1.96%) compared to FeSO4 salt (28.04±0.50%). The synthesis of ferritin by the Caco-2 cells was significantly increased (p <0.05) after absorbing iron from EPSKar1-iron complex (32.72±0.73 ng/mL) than from only FeSO4 salt (21.77±0.10 ng/mL). Further, the EPSKar1-iron complex was experimented on anaemic rats for a period of 20 days to evaluate iron bioavailability after inducing anaemia by feeding iron free diet for 62 days. The apparent digestibility coefficient (ADC), % iron balance, and % retention/intake was found to be significantly higher (p <0.05) in rats fed with the highest concentration of complex (50 mg/Kg BW), when compared to only FeSO4 fed rats and the rats fed with 25 mg EPSKar1-iron complex/Kg BW. These results were consistent throughout the experimental period of 20 days. There was a significant increase (p <0.05) in the blood haemoglobin content (12.76±0.19 g/dL) of rats fed with higher concentration of complex when compared to rats fed with low concentration of complex (11.91±0.29 g/dL) and only FeSO4 fed rats (10.54±0.21 g/dL). The iron absorption indicators like transferrin and ferritin were found to be significantly higher (p <0.05) in high concentration complex fed rats when compared with only FeSO4 and low concentration complex fed rats. Taken together, these findings suggest that the iron in EPSKar1 complex form has greater bioavailability than uncomplexed iron. Hence, the complex may serve as an ideal molecule for fortifying food products as a source of iron fortificant. Nevertheless, further human clinical trials are highly warranted in order to validate it as an effective nutraceutical for an anaemic group of population.
