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1. Introduction
Pumpkin belongs to the Cucurbitaceae family and is broadly grown around the warmer regions across the world. The worldwide production of pumpkin, squash, and gourd was 27 million tons (MT) in 2017. India is the second-largest producer, with about 20% (5.56 MT) of the total worldwide production [1]. Pumpkin contains flesh (72–76%), by-product peel (2.6–16%), and seeds (3.2–4.4%) [2]. The pumpkin processing industry separates useful parts of the fruit from by-products, which constitute peel and seeds [3]. Pumpkin seeds are a vital source of bioactive components such as fats, essential proteins, vitamins, phytosterols, squalene, tocopherols, and carotenoid pigments [4, 5]. It is consumed as snack food in several countries, and its oil is generally used in cooking and for pharmaceutical applications. The pumpkin seeds have high moisture; therefore, they are highly susceptible to microbial spoilage and chemical alteration [6].
The demand for pumpkin seeds is increasing on account of their high nutritional and functional properties. This is likely to encourage investigations on the effects of postharvest processing and industrial by-product utilization to prevent degradation of quality. Drying is the most appropriate technique to stop microbial spoilage and to prolong the storage of such seeds [7]. Furthermore, it also reduces transportation and storage costs. However, physical and nutritional changes can occur in food materials including some quality characteristics, such as texture, color, and flavor [8]. Thus, the selection of proper drying techniques is a particularly important step in food processing.
A mechanical vibration-assisted fluidized bed is also a potential advanced drying technique to improve the fluidization of such particles [9]. The electrical energy consumed by blower and vibration motor in a vibro-fluidized bed dryer (VFBD) has been reported to be around 55% of the electrical energy of the blower in FBD [10]. Furthermore, by inducing vibration to the fluidized bed, homogeneous temperature circulation can be achieved [11], agglomeration of solid particles can be prevented and the fluidization velocity and pressure drop can be reduced [12, 13].
In drying, heat and mass transfer take place at the same time due to its complexity. Therefore, optimization of drying process conditions is important, and for this, mathematical modeling is an exceptional tool [14]. Moreover, modeling is applied to find out the drying time and general nature of drying. To design and select dryers, knowledge of drying behavior is very important. There are several studies reported in the literature to predict or develop the most suitable mathematical model for drying kinetics. Generally, to predict the drying behavior of agricultural commodities different mathematical models, namely, empirical, semiempirical, and theoretical, are used. In most of the drying experiments, empirical models validate the exceptional fitting of data but overlook the basics of the drying processes. While theoretical models justify diffusion mechanism or heat and mass transfer mechanism [15], there are several investigations on mathematical modeling of pumpkin seeds drying kinetics for natural and forced convection solar dryer [6], solar tunnel dryer [16], tray drying [17], fluidized bed drying [17–20], and infrared dryer [21]. However, no study was found for pumpkin seed drying using a vibro-fluidized bed dryer.
The drying of biological samples is a complex method and on occasions, may not be explained using traditional models. For that purpose, a neural network approach was also attempted for this study considering the novel nature of the drying process. Artificial neural networks (ANN) are a machine learning method applied to predict intricate correlations among input and output conditions of drying and nonlinear problems [22]. Many researchers used ANN to explain moisture content, rate of drying, moisture ratio, effective diffusivity, and specific energy consumption for different agriculture products [23–25]. Alvarez et al. [26] successfully used ANN to correlate the heat and mass transfer parameters and vibration parameters (amplitude, frequency, and vertical motion) for poppy seeds dried in a vibrated fluidized bed dryer. Kumar et al. [27] used a feedforward backpropagation ANN model to predict the moisture ratio by feeding drying temperature and time as input parameters. According to our intensive literature search, no work has been reported on the modeling of drying kinetics of pumpkin seeds in a VFBD by ANN and its comparison with semiempirical drying models to the best of our knowledge. Moreover, information on the mass transfer parameters for pumpkin seed drying using VFBD is not available. Such parameters are useful in designing drying equipment and optimizing industrial drying processes and hence were taken up in this investigation.
Therefore, the aim of the current work is to perform pumpkin seeds drying in a VFBD at various drying parameters along with the determination of drying and mass transfer parameters. The study also deals with the prediction and comparison of the drying behavior of pumpkin seeds using semiempirical modeling and ANN.
2. Materials and Methods
2.1. Sample Preparation
Pumpkins (Cucurbita maxima) were procured from the market of Narela, New Delhi, India, during the summer season. The pumpkins were cut in half followed by manual removal of the seeds. Undeveloped seeds were removed and the seeds exhibiting the same size were washed with deionized water and pressed in a sieve, to remove the stringy pulp. The washed seeds were stored in low-density polyethylene (LDPE) airtight bags at −10°C and were subjected to drying treatment within 24 h of storage. Hot-air oven method was used to estimate initial moisture content at 105°C for 24 h [28]. All readings were replicated thrice for accuracy.
2.2. Experimental Setup
An electrical power-operated vibro-fluidized bed dryer was fabricated at the Department of Food Engineering, NIFTEM, Haryana, India, as depicted in Figure 1. The components and specifications of VFBD are presented in Table 1.
[figure omitted; refer to PDF]
Table 1
Design specifications of the vibro-fluidized bed dryer.
Components | Specifications |
Drying chamber | Dimension = 0.35 m × 0.40 m (diameter × height) |
Volume = 0.0384 m3 | |
Capacity = 10–15 kg per drying cycle | |
Plenum chamber | Dimension = 0.35 m × 0.25 m (diameter × height) |
Distributor plate | Dimension = 0.35 m diameter and 0.0002 m aperture diameter |
Vibration motor (DC) with eccentric wheel blower | 1HP |
2HP with 2800 rpm-3 phases | |
Air flow rate = 650–670 m3/h | |
Variable frequency drive (delta: VFD015S21D, US) | 1.5 kW-1 phase |
Frequency = 0–60 Hz | |
Heating chamber | 0.50 × 0.50 × 0.40 m |
Heater | 1 coil type = 2 kW and 4 fins type = 4 kW |
PID controller (Instron IN- 312 and KRITMAN GHC-100, India) | 30°C to 600°C |
Temperature measurement | PT-100 and k-type thermocouple |
Hot-air anemometer (Mextech AM 4208, Mextech, India) | Sensor = 0.0072 m |
Air velocity range = 0.4–45 m/s | |
Air temperature range = 0–50°C | |
Air flow = 0–9999 m3/min |
2.3. Vibration Intensity
The vibration intensity (Γ) was calculated using the following equation [29]:
2.4. Drying Experiments
The experiments were conducted at different conditions as shown in Table 2. Air velocity selected for fluidization was 4 m/s as given by Kaveh et al. [7] for squash seeds. Before each experiment, the three dryers were run for 30 min in no-load condition to accomplish the chosen operating parameters of the drying chamber. For drying, pumpkin seeds of approximately 200 g were placed on the perforated sieve of the VFBD. The weight of moisture was recorded at 5 min intervals. Drying was continued until 0.007–0.009 kg H2O/kg dm moisture remained in the sample, which is the safe storage moisture content for high-oil-content seeds. The dried samples were cooled in desiccators at an ambient temperature (27°C) for 15 min and then filled in LDPE bags for additional investigations.
Table 2
Experimental parameters for VFBD.
Operating parameters | VFBD |
Initial moisture content (kg H2O/kg dry matter) | 1.442 ± 0.082 |
Air temperature (°C) | 40, 50, and 60 |
Air velocity (m/s) | 4 |
Vibration intensity | 4.29 (A = 10 mm and |
Bed height (cm) | Thin layer |
Ambient temperature (°C) | 27–30 |
Relative humidity (%) | 45–50 |
2.5. Drying Kinetics and Mathematical Modeling
The moisture ratio during drying was computed using the standard equation (Babar et al., 2020).
The experimental data of drying of pumpkin seeds were fitted to six thin-layer drying models as shown in Table 3. The parameters of the models were estimated using a nonlinear regression procedure. For selecting the most appropriate model to depict thin-layer drying, determination coefficient (R2), chi-square (χ2), and mean square error (MSE) were used. These parameters were calculated using standard equations [18, 31].
Table 3
Most commonly used semiempirical models for illustrating the drying kinetics of seeds.
Models | Model equation |
Page | |
Modified page | |
Newton | |
Henderson and Pabis | |
Two-term | MR = a |
Logarithmic | MR = a |
2.6. ANN Modeling
ANN mimics the functionality of a neuron in the human brain. It adapts to new knowledge and identifies patterns in fuzzy and inaccurate data. In this work, the modeling of experimental data was also done by a multilayer FFBP neural network using the neural network toolbox of MATLAB v.2012a (MathWorks Inc., USA). The ANN model was provided with different air temperature and drying time data as input, while MC and MR were given as output (supervised training) with one hidden layer, to reduce the complexity of the model. The best ANN infrastructure for accurate drying data prediction was done on a trial-and-error basis. There were numerous backpropagation training algorithms such as scaled conjugate gradient (SCG), Polak–Ribière conjugate gradient (CGP), BFGS quasi-Newton (BGF), Levenberg–Marquardt (LM), resilient (Rprop), gradient descent (GDX) backpropagation, etc. [32]. From these, the Levenberg–Marquardt training algorithm (TRAINLM) was used because of its previously proven efficiency in drying data prediction [33].
For model development, 70% was used for training and 30% of the remaining data for testing and validation, respectively. The TANSIGMOID activation function was exercised to correlate the signal of input and output of each layer and to decrease the processing time. As proposed by Dorofki et al. [34], as an approximation function, the PURELIN transfer function was applied to the output layer. The number of iterations was limited to 1000 to reduce the validation time while ensuring proper model termination (attainment of error goals). As for performance indicators of the model, MSE and correlation coefficient (R) were recorded.
The signal received to the first HL from the input layer was expressed as follows [22]:
The output signal of neuron can be expressed using the following equations, respectively:
Output layer will receive 10 signals from 10 HL, which was expressed using the following equation:
PURELIN function was employed to the receiving neurons of the output layer to obtain the outcome of the ANN model, as follows:
The error between target and output of the selected backpropagation model was estimated and minimized to an acceptable level by adjusting the weights of the structure.
2.7. Estimation of Effective Moisture Diffusivity (Deff) and Activation Energy (Ea)
For the valuation of Deff, slab geometry was considered for pumpkin seeds, owing to its small thickness as related to other dimensions [35]. An assumption was made that negligible shrinkage due to drying took place with uniform distribution of initial MC. Fick’s law was used to calculate Deff of pumpkin seeds [16], which was linearized as suggested by Golpour et al. [36]. Activation energy was determined using Arrhenius function, which expressed the relationship of Deff and temperature.
2.8. Mass Transfer Parameters
The parameters such as Biot number (Bi) and mass transfer coefficient (hm) were determined by the equation given by Dincer and Hussain [37]. Bi, a dimensionless number, represents the correlation between internal and external convective heat transfer [38]. It also specifies the resistance of moisture transfer inside a product and is affected by the drying medium and product type. Bi was calculated as
The relationship between air velocity and heating and cooling coefficient is provided by the Dincer number (Di). Di was expressed using
Additionally, the convective mass transfer coefficient was determined using the following equation:
3. Results and Discussion
3.1. Drying Characteristics
The initial moisture content of pumpkin seeds was determined as 1.52 ± 0.013 kg H2O/kg dry matter which reduced to 0.08 ± 0.01 kg H2O/kg dry matter. The drying curves of dried pumpkin seeds in VFBD at different temperatures are shown in Figure 2. The reduction of moisture content of pumpkin seeds with time was observed at all drying conditions, as shown in Figure 2(a). A decrease in moisture content showed that diffusion mechanism was regulated by mass transfer within seeds (Ashtiani et al.) [39]. From Figure 2(b), it was observed that there was no constant rate period; this might be due to the thin-layer drying of pumpkin seed, which was unable to deliver a constant supply of moisture while drying. As drying proceeds, the free surface moisture evaporates and when surface moisture is insufficient, moisture diffusion takes place from the core of the seeds to the surface which also explained the decrease in MR, as observed in Figure 2(c). Therefore, a reduction of drying rate was observed with time with the initiation of falling rate. Another possible reason for faster moisture diffusion from pumpkin seeds could be its flat geometry exhibiting large surface-to-volume ratio. The migration of water from larger surfaces to its surrounding is much faster than in smaller surface seeds.
[figures omitted; refer to PDF]
The drying rate was found to increase from 0.0063 to 0.0091 kg H2O/kg dm. min with the rise in temperature from 40 to 60°C. It showed that elevated temperature elevates heat transfer gradient between surrounding air and seeds, which stimulates a higher drying rate and subsequently reduces drying time [40]. Similar results of drying curves were found in castor oil seeds (Ricinus communis) [31], watermelon seeds [41], pumpkin seeds [17], Jatropha curcas L. seeds [42], and garlic clove [43, 44]. Stakić and Urošević [45] dried poppy seeds in VFBD and reported that with the rise of temperature, the drying rate improved and drying time reduced. Lima et al. [46] also concluded that proper moisture evaporation and homogeneity were obtained in vibro-fluidized bed drying than conventional fluidized bed drying.
3.2. Mathematical Modeling
The results of experimental data were fitted to semiempirical models and the model coefficients were determined and are presented in Table 4. The best-fitted model was selected based on the maximum R2 and lowest value of MSE and χ2. The range of R2, MSE, and χ2 for all experiments were 0.9923 to 0.9990, 7.55 × 10−5 to 7.77 × 10−4, and 8.48 × 10−5 to 6.48 × 10−4, respectively. The experimental data was fitted well in all the models with R2 greater than 0.99. But the two-term model was fitted exceptionally well with the highest R2 − 0.999, and lowest χ2 − 1.03 × 10−4 and MSE 7.55 × 10−5 at a temperature of 60°C. The validation of the best-fit model (i.e., two-term) was also conducted using MATLAB curve fitting tool. The goodness of validation as predicted parameters are SSE: 0.00113 and RMSE: 0.00870. It was observed that drying rate constant (k) was significantly (
Table 4
Statistical parameters and the coefficients of pumpkin seeds dried in VFBD.
Model | Temperature, °C | Empirical constants | R2 | MSE | χ2 |
Two-term | 40 | k = 0.0397 ± 0.0049, a = 0.4842 ± 0.0048 | 0.9967 | 2.66 × 10−4 | 2.72 × 10−4 |
b = 0.4842 ± 0.0049, k0 = 0.0396 ± 0.0013 | |||||
50 | k = 0.0614 ± 0.0009, a = 0.5021 ± 0.0078 | 0.9949 | 4.88 × 10−4 | 6.27 × 10−4 | |
b = 0.5021 ± 0.0078, k0 = 0.0614 ± 0.0008 | |||||
60 | k = 0.4405 ± 0.3801, a = 0.1014 ± 0.0373 | 0.9990 | 7.55 × 10−5 | 1.03 × 10−4 | |
b = 0.8968 ± 0.0.036, k0 = 0.0594 ± 0.0019 | |||||
Newton | 40 | k = 0.0410 ± 6.63 × 10−4 | 0.9956 | 3.00 × 10−4 | 3.14 × 10−4 |
50 | k = 0.0611 ± 0.0015 | 0.9939 | 4.89 × 10−4 | 5.18 × 10−4 | |
60 | k = 0.0657 ± 0.0013 | 0.9964 | 2.80 × 10−4 | 3.00 × 10−4 | |
Logarithmic | 40 | k = 0.0416 ± 0.0012, a = 0.9637 ± 0.0011 | 0.9972 | 2.36 × 10−4 | 2.17 × 10−4 |
b = 0.0136 ± 0.0059 | |||||
50 | k = 0.0592 ± 0.0027, a = 1.010 ± 0.0215 | 0.9943 | 4.51 × 10−4 | 5.41 × 10−4 | |
b = −0.0109 ± 0.0101 | |||||
60 | k = 0.06430 ± 0.0025, a = 0.9746 ± 0.015 | 0.9971 | 2.25 × 10−4 | 2.81 × 10−4 | |
b = 0.0012 ± 0.0088 | |||||
Modified page | 40 | k = 0.0252 ± 3.38 × 10−4n = 1 ± 0 | 0.9956 | 3.01 × 10−4 | 3.28 × 10−4 |
50 | k = 0.0306 ± 7.96 × 10−4n = 1 ± 0 | 0.9939 | 4.88 × 10−4 | 5.50 × 10−4 | |
60 | k = 0.0329 ± 6.81 × 10−4n = 1 ± 0 | 0.9964 | 2.79 × 10−4 | 3.23 × 10−4 | |
Page | 40 | k = 0.0578 ± 0.0025, n = 0.9000 ± 0.012 | 0.9989 | 7.77 × 10−5 | 8.48 × 10−5 |
50 | k = 0.0503 ± 0.007, n = 1.064 ± 0.044 | 0.9947 | 4.24 × 10−4 | 4.77 × 10−4 | |
60 | k = 0.0856 ± 0.0052, n = 0.9111 ± 0.0021 | 0.9986 | 1.13 × 10−4 | 1.30 × 10−4 | |
Henderson and Pabis | 40 | k = 0.0397 ± 7.49 × 10−4a = 0.9683 ± 0.011 | 0.9967 | 2.25 × 10−4 | 2.48 × 10−4 |
50 | k = 0.0615 ± 0.0019, a = 1.0046 ± 0.0197 | 0.9939 | 5.26 × 10−4 | 5.49 × 10−4 | |
60 | k = 0.0641 ± 0.015, a = 0.9752 ± 0.015 | 0.9971 | 2.36 × 10−4 | 2.59 × 10−4 |
[figures omitted; refer to PDF]
3.3. ANN Performance
ANN model was successfully trained by TRAINLM algorithm with 10 neurons in the HL. The correlation coefficients (R) of ANN model for training, testing, and validation were 0.9999, 0.9997, and 0.9999, respectively, with a mean square error of 2.12 × 10−4 as shown in Figure 4. The weights and bias of the final ANN topography for pumpkin seed drying in VFBD are shown in Table 5.
[figure omitted; refer to PDF]Table 5
Weights and biases for the obtained neural network.
j, k | Weights | Bias | ||||
bj | bk | |||||
1 | 3.4253 | −2.6199 | 0.3718 | 0.3905 | −4.959 | 2.1745 |
2 | 3.3791 | 5.0788 | 0.8209 | 0.8345 | −5.0407 | 2.2252 |
3 | −5.4467 | 6.1391 | −0.61 | −0.6045 | 3.7684 | — |
4 | 4.4125 | −2.2221 | −0.6507 | −0.6479 | −1.7083 | — |
5 | 0.5999 | 3.3198 | −0.0667 | −0.0666 | −1.105 | — |
6 | −5.3801 | −3.3426 | 0.2788 | 0.2695 | −0.8644 | — |
7 | −4.0007 | −1.7235 | −0.2726 | −0.2522 | −0.8594 | — |
8 | 0.1321 | 4.0858 | −3.5083 | −3.5636 | 4.7189 | — |
9 | 3.6053 | 2.1813 | −0.4538 | −0.4614 | 4.7392 | — |
10 | 1.7939 | 4.7844 | 2.0048 | 2.0376 | 7.2173 | — |
The predicted moisture content (R2 = 0.9967; MSE = 5.29 × 10−5) and moisture ratio (R2 = 0.9963; MSE = 2.42 × 410−5) of optimum network structure (2-10-2 topology) was projected by TANSIGMOID as shown in Figure 5. Kaveh [35] found that the ANN model-FFBP provided the best extrapolation of moisture ratio (R2 = 0.9944) of squash seeds dried in an FBD. Jafari et al. [23] determined that FFBP network with TRAINLM, TANSIGMOID activation function, and 2-5-1 topology showed R2 of 0.9996 and minimum MSE of 3.94 × 10−5 for predicting the moisture ratio of onion dried in fluidized bed dryer. The reported work was similar to our findings for pumpkin seeds.
[figure omitted; refer to PDF]3.4. Comparative Assessment of Semiempirical and Artificial Neural Network Model
Two-term model was found to most adequately fit the experimental data with maximum R2 and minimum MSE. Subsequently, the two-term model was selected for comparison with the ANN model. The comparative statistical indicators are presented in Table 6. ANN provided a relatively reasonable fit at all drying conditions, which was in the range of the maximum R2 and minimum MSE values of the semiempirical model. This illustrates that ANN is more capable of correlating all external and internal variables and is more likely to provide accurate predictions at other unseen processing conditions [22, 23].
Table 6
Comparative analysis of semiempirical and ANN model.
Model | R2 | MSE |
Two-term | 0.9949–0.9990 | 1.03×10−4–6.27 × 10−4 |
ANN | 0.9963 | 2.42 × 10−5 |
3.5. Moisture Diffusivity and Activation Energy
The Deff was estimated from the slope obtained from the graph presented in Figure 6(a) for VFBD. The values for the same are displayed in Table 7. The outcomes illustrated that Deff values of pumpkin seeds changed from 1.12 va10−9 ± 3.62 .610−10 to 2.28 to10−9 ± 1.43 .410−10 m2/s in the temperature range of 40 to 60°C in VFBD, respectively. The values attained in this analysis were within the range of food materials, which is usually in the range of 10−11 to 10−9 m2/s [47].
[figures omitted; refer to PDF]
Table 7
Deff, Ea, and mass transfer constraints of pumpkin seeds at different drying temperatures.
Temperature (°C) | Deff (m2/s) | Ea (kJ/mol) | k | Bi | hm (m/s) |
40 | 1.12 ± 3.62 × 10−10 | 32.71 ± 1.05 | 0.04 ± 0.00 | 0.51 ± 0.01 | 1.49 ± 4.89 × 10−8 |
50 | 1.55 ± 6.7 × 10−11 | 0.064 ± 0.00 | 0.60 ± 0.01 | 2.42 ± 9.24 × 10−8 | |
60 | 2.28 ± 1.43 × 10−10 | 0.44 ± 0.01 | 1.26 ± 0.01 | 7.45 ± 5.52 × 10−8 |
It was observed that the moisture diffusivity significantly (
The graphical representation of logarithmic moisture ratio ln Deff versus T−1 for drying of pumpkin seed at the different temperatures is shown in Figure 6(b) for VFBD. Ea of pumpkin seeds was determined as 32.71 ± 1.05 kJ/mol in VFBD. According to Bezerra et al. [49], if Ea is greater, the evaporation of moisture from the surface of the sample is slower due to strongly bonded moisture with other compounds. The value of Ea found in this report was within the range of Ea obtained for food products and low as compared to values reported in previous studies [18, 19].
3.6. Mass Transfer Parameters
Bi represents the rate of moisture diffusion during the process of drying. The Bi and hm values are summarized in Table 7. As expected, the Bi number and hm were significantly (
4. Conclusions
Pumpkin seeds were dried in a VFBD at 40, 50, and 60°C at a constant air velocity of 4 m/s with 4.26 (A = 10 mm and
Acknowledgments
The authors are thankful to members of the Department of Food Engineering, NIFTEM (Sonipat), for their facilities, support, and encouragement. The first author thanks the Ministry of Tribal Affairs, Government of India, for providing fellowship vide Grant no. 201718-NFST-MAH-02452.
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Abstract
Pumpkin seeds were dried in a vibro-fluidized bed dryer (VFBD) at different temperatures at optimized vibration intensity of 4.26 and 4 m/s air velocity. The drying characteristics were mapped employing semiempirical models and Artificial Neural Network (ANN). Prediction of drying behavior of pumpkin seeds was done using semiempirical models, of which, one was preferred as it indicated the best statistical indicators. Two-term model showed the best fit of data with R2 − 0.999, and lowest χ2 − 1.03 × 10−4 and MSE 7.55 × 10−5. A feedforward backpropagation ANN model was trained by the Levenberg–Marquardt training algorithm using a TANSIGMOID activation function with 2-10-2 topology. Performance assessment of ANN showed better prediction of drying behavior with R2 = 0.9967 and MSE = 5.21 × 10−5 for moisture content, and R2 = 0.9963 and MSE = 2.42 × 10−5 for moisture ratio than mathematical models. In general, the prediction of drying kinetics and other drying parameters was more precise in the ANN technique as compared to semiempirical models. The diffusion coefficient, Biot number, and hm increased from 1.12 × 10−9 ± 3.62 × 10−10 to 1.98 × 10−9 ± 4.61 × 10−10 m2/s, 0.51 ± 0.01 to 0.60 ± 0.01, and 1.49 × 10−7 ± 4.89 × 10−8 to 3.10 × 10−7 ± 7.13 × 10−8 m/s, respectively, as temperature elevated from 40 to 60°C. Arrhenius’s equation was used to the obtain the activation energy of 32.71 ± 1.05 kJ/mol.
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1 Department of Food Engineering, National Institute of Food Technology Entrepreneurship and Management, Kundli, Sonipat 131 028, Haryana, India
2 Department of Food Engineering, National Institute of Food Technology Entrepreneurship and Management, Kundli, Sonipat 131 028, Haryana, India; Livestock Production and Management Section, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly 243 122, Uttar Pradesh, India