Encapsulation of Lactiplantibacillus plantarum CRD7 in sub-micron pullulan fibres by spray drying: Maximizing viability with prebiotic and thermal protectants.

Pullulan was used as the wall material for microencapsulation of L. plantarum CRD7 by spray drying, while isomalto-oligosaccharides (IMO) was used as prebiotic. Also, the effect of different thermal protectants on survival rate during microencapsulation was evaluated. Taguchi orthogonal array design showed that pullulan at 14 % concentration, IMO at 30 % concentration and whey protein isolate at 20 % rate were the optimized wall material, prebiotic and thermal protectant, respectively for microencapsulation of L. plantarum. FESEM images revealed that the spray-dried encapsulates were fibrous similar to those produce by electrospinning, while fluorescence microscopy ascertained that most of the probiotic cells were alive and intact after microencapsulation. The adsorption-desorption isotherm was of Type II and the encapsulate had specific surface area of 1.92 m2/g and mean pore diameter of 15.12 nm. The typical amide II and III bands of the bacterial proteins were absent in the FTIR spectra, suggestive of adequate encapsulation. DSC thermogram showed shifting of melting peaks to wider temperature range due to interactions between the probiotic and wall materials. IMO at 30 % (w/w) along with WPI at 20 % concentration provided the highest storage stability and the lowest rate of cell death of L. plantarum after microencapsulation. Acid and bile salt tolerance results confirmed that microencapsulated L. plantarum could sustain the harsh GI conditions with >7.5 log CFU/g viability. After microencapsulation, L. plantarum also possessed the ability to ferment milk into curd with pH of 4.62.

Fabrication and characterization of electrospun catechins-loaded nanofibres for fortification of milk.

Catechins in their free form are bitter in taste, and undergo deterioration and oxidation during processing and storage that limit their use as nutraceuticals in foods. Therefore, catechins were electrospun using zein as encapsulating polymer into nanofibres at 15, 18 and 21% w/w concentrations, 16, 20 and 24 kV applied voltage and 0.5 and 1.0 mL/h feed rate. The electrospinning conditions were optimized using Taguchi L18 (21 × 32) design. Encapsulation efficiency as high as 92.8% and mean fibre diameter as low as 95.2 nm were obtained at 18% concentration of zein, 0.5 mL/h feed rate and 20 kV applied voltage. Scanning electron and atomic force micrographs revealed that the nanofibres produced at zein concentration of 18% and above were clean and beadfree, with cylindrical morphology and non-porous topography. The hydrodynamic diameter, zeta potential and polydispersity index of catechins-loaded nanofibres at optimized conditions were 172.3 nm, -26.3 mV and 0.15. FTIR spectroscopy and X-ray diffractometry confirmed that catechins were encapsulated within the nanofibres. The catechins got released from loaded nanofibres in a controlled and sustained manner, while their antioxidant property was retained. The physico-chemical and sensory qualities of milk were not affected after fortification with catechins-loaded nanofibres. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05891-0.

Characterization of withanolides and bacoside A-loaded proniosomes: effect on oxidative stress and survival under hypergravity in rodent model.

OBJECTIVE: This work provides characterization of withanolides and bacoside A proniosomes, and evaluating their potency in rat model for combating oxidative stress-induced blood-brain barrier (BBB) damage and their survival under hypergravity. SIGNIFICANCE: The delivery system was aimed for sustained drug release in plasma and brain, which could improve their efficiency and provide a therapeutic approach to combat oxidative damage and restore BBB integrity. METHODS: Proniosomes were prepared using withanolides extracted from the roots of W. somnifera and bacoside A derived from the leaf extract of B. monnieri by thin film hydration technique. In vitro release of withanolides and bacoside A from the proniosomes was studied. In vivo experiments were conducted in Wistar Albino rat model to evaluate the efficacy of drug-loaded proniosomes in improving the antioxidant activity in plasma and brain, restoring BBB integrity and combating hypergravity conditions. RESULTS: The withanolides and bacoside A-loaded proniosomes showed slow and sustained release of just 62.0 ± 2.87 and 62.9 ± 3.41%, respectively, in 9 h period against the release of 98-99% for the extracts that served as control. Trials conducted in vivo revealed a significant (p < .05) increase in the activity of antioxidant enzymes in both plasma and brain. Also, minimal extravasation of Evans blue dye into the brain (15 ± 0.03 and 16 ± 0.03 ng/g in treated groups against 110 ± 0.01 ng/g in control) of the rats fed with drug-loaded proniosomes was indicative of minimal damage to BBB. Rats fed with drug-loaded proniosomes survived to the extent of 75-83.3% against simulated hypergravity as compared to the control group in which only 50% survived. CONCLUSION: Proniosomes provided sustained release of drugs, which helped to protect BBB integrity, thereby combating hypergravity.

In vitro safety assessment of electrohydrodynamically encapsulated Lactiplantibacillus plantarum CRD7 and Lacticaseibacillus rhamnosus CRD11 for probiotics use.

The current study aimed to validate the safety of electrohydrodynamically encapsulated Lactiplantibacillus plantarum CRD7 and Lacticaseibacillus rhamnosus CRD11 in accordance with guidelines of FAO/WHO and ICMR/DBT. In vitro assays such as mucin degradation, hemolysis of blood cells, antimicrobial susceptibility pattern, possession of virulence factors, biogenic amine, and ammonia production were assessed. In results, the cross-streak and co-culture techniques revealed that CRD7 and CRD11 were compatible in vitro. Upon visual inspection through scanning electron and fluorescence microscopy, the integrity of bacterial cell membrane was confirmed even after the encapsulation process. CRD7 and CRD11 were non-hemolytic and showed negative responses to gelatinase, urease, and DNase activities. Non-mucinolytic activity of CRD7 and CRD11 was verified by measuring cell growth rate (p < 0.05) in different modified media followed by SDS-PAGE. High-performance liquid chromatography analysis revealed that both the strains did not produce biogenic amines (putrescine, cadaverine, histamine, and tyramine). Neither of the Lactobacillus strains produced ammonia after growing in BHI broth for 5 days at 37 °C. L-lactate production was highest (p < 0.05) in CRD11 (8.83 g/L), followed by CRD7 (8.16 g/L), whereas the lowest (p < 0.05) D-lactate was registered for encapsulated CRD11 (0.33 g/L) and CRD7 (0.49 g/L). The antibiogram profile determined through minimum inhibitory concentration showed that CRD7 and CRD11 were sensitive to key antibiotics suggested by EFSA except for gentamycin and kanamycin. PCR data on virulence genes demonstrated that both strains were safe for probiotic use. Moreover, CRD7 and CRD11 strains caused insignificant (p > 0.05) changes in the cell viability of Caco-2 cells as estimated by MTT (98.94-99.50%) and NR uptake (95.42-97.03%) assays and showed sensitivity to human serum. According to the results of these evaluated attributes, it is concluded that L. plantarum CRD7 and L. rhamnosus CRD11 are safe, non-toxic to human epithelial cells, and thus may be potentially suitable for various food/feed applications.

Microencapsulation of curcumin by spray drying: Characterization and fortification of milk.

Curcumin, the major bioactive component of turmeric (Curcuma longa), was microencapsulated by spray drying in the matrix of HI-CAP 100 (resistant starch)/ maltodextrin and whey protein isolate to improve its oral bioavailability and solubility. Taguchi orthogonal array design (L18) was used to optimize the spray drying conditions. The optimal conditions for microencapsulation were inlet drying air temperature of 185 °C, feed rate of 6 mL/min and HI-CAP 100 as wall material. The moisture content, encapsulation efficiency and bulk density at these conditions were 4.65%, 82.42% and 358.40 kg/m3, respectively. The spray-dried microcapsules were spherical-shaped with folds and vacuoles. The yellowness index and a* value of curcumin decreased after microencapsulation. FTIR spectroscopy indicated that the curcumin after microencapsulation presumably retained its chemical structure. DSC thermograms confirmed that the microcapsules were heat stable up to 200 °C. The microcapsules had better heat stability and sustained in-vitro release as compared to that of pure curcumin. The DPPH free radical scavenging activity of curcumin was 61.43%, which was largely unaffected after microencapsulation. Fortification of milk with HI-CAP 100-based microcapsules at the selected dose had no adverse effect on organoleptic properties as compared to normal milk. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05142-0.

Nanoencapsulation of casein-derived peptides within electrospun nanofibres.

BACKGROUND: Bioactive peptides derived from milk proteins are recognized as functional foods, but their consumption is limited by undesirable or bitter flavour, poor stability, and low bioavailability. Electrospinning is a versatile process for encapsulation of various bioactive compounds in the form of nanosized fibres, which can circumvent these disadvantages. This study was aimed at the preparation of casein-derived peptides-loaded nanofibres through electrospinning and characterizing them for fortification of milk. RESULTS: Pullulan at 100, 120, and 140 g kg-1 concentrations was used for electrospinning of peptides. Scanning electron and atomic force micrographs revealed the formation of clean bead-free peptides-loaded pullulan nanofibres at 120 and 140 g kg-1 concentrations with mean diameter of 60.45-133.05 nm and encapsulation efficiency of 72.95-86.04%. Fourier transform infrared spectra and X-ray diffractograms revealed the absence of interactions between the functional groups of pullulan and peptides during electrospinning. The zeta potential of the peptides-loaded nanofibres ranged from -15.6 to -24.6 mV, and the hydrodynamic diameter varied from 118.7 to 256.2 nm. The peptides from electrospun nanofibres showed sustained release to the extent of 75.3% after 8 h in gastrointestinal pH conditions. The release kinetics of peptides from nanofibres was best fitted to a Peppas-Sahlin model (R2  = 0.987), and through diffusion and erosion mechanisms. The antioxidant activity of pure peptides and those from nanofibres was comparable. The physico-chemical qualities of milk fortified with encapsulated peptides did not show noticeable difference either. CONCLUSIONS: From the morphological, ultrastructural, particle size, encapsulation efficiency, release kinetics, and antioxidant activity data, it was inferred that electrospinning could be an effective technique for nanoencapsulation of casein-derived bioactive peptides. These peptides-loaded nanofibres could be used for fortification of milk and milk products. © 2021 Society of Chemical Industry.

Formulation and characterization of catechin-loaded proniosomes for food fortification.

BACKGROUND: To overcome the problems associated with niosomes, proniosomes - a dry powder - was prepared to nanoencapsulate catechins using Span 60 as surfactant, cholesterol as stabilizer and maltodextrin, lactose monohydrate and pullulan as wall materials. The proniosomes were made by the thin-film hydration technique, and were characterized for fortification in milk beverages. Scanning and atomic force microscopic images showed the varying morphology and ultrastructure of the proniosomes. The mean hydrodynamic diameter of 193.57-262.52 nm, polydispersity index of 0.24-0.25 and zeta potential of -15.8 to -24.73 were suggestive of the size, homogeneity and stability of the catechin-loaded proniosomes. X-ray powder diffractograms and Fourier transform infrared (FTIR) spectra provided insight about the interaction between catechins and wall materials. Entrapment efficiency and in vitro release were calculated to determine the extent of nanoencapsulation of catechins and their bioavailability, respectively. The nanoencapsulates were fortified in milk and yogurt to find their organoleptic acceptability. RESULTS: Moisture content was found to be 20-30 g kg-1 , indicating longer stability of the proniosomes. Scanning electron microscopic and atomic force microscopic images revealed the ultrastructure and spherical-shaped morphology of proniosomes. Entrapment efficiency of catechins using pullulan as wall material was as high as 83.43%. In vitro release studies revealed the sustained release of catechins from the proniosomes. FTIR and X-ray diffraction spectra revealed the absence of chemical interactions between catechins and encapsulants. CONCLUSION: Food-grade proniosomes are a good vehicle for fortification of milk and yogurt without noticeable adverse changes in their organoleptic and physicochemical properties, thus increasing the potential for bioavailability of catechins in the gastrointestinal tract. © 2020 Society of Chemical Industry.

Monitoring foot surface temperature using infrared thermal imaging for assessment of hoof health status in cattle: A review.

Detection of lameness early in cows is important from the animal welfare point of view and for reducing economic losses. Currently, many studies are being conducted for assessment of hoof health status by measuring the surface temperature of skin in cattle and other animal species in different parts of the world. Infrared Thermography (IRT) is able to detect lesions of hooves associated with lameness by measuring the changes in coronary band and hoof skin surface temperature. The surface temperature of a lame limb will be increased when the hoof has lesion(s). IRT has been used as a non-invasive diagnostic tool for early detection of hoof lesions based on the temperature difference between affected and non-affected hoof and maximum foot temperature on the regions of interest. In spite of having many potential applications in cattle production, factors affecting the temperature readings in thermograms must also are considered while taking images. Standard operating procedures must be established before taking thermographs under different circumstances, by considering all the factors that affect its normal function. IRT may help in minimising the cost of veterinary services, low yield, compromised fertility and culling expenses, where lameness cannot be resolved in early stages.

Infrared thermography: A potential noninvasive tool to monitor udder health status in dairy cows.

The animal husbandry and livestock sectors play a major role in the rural economy, especially for the small and marginal farmers. India has the largest livestock population in the world and ranks first in the milk production. Mastitis is the most common and expensive infectious disease in dairy cattle. The global economic losses per year due to mastitis amounts to USD 35 billion and for Indian dairy industry ₹6000 crores per year. Early detection of mastitis is very important to reduce the economic loss to the dairy farmers and dairy industry. Automated methods for early and reliable detection of mastitis are currently in focus under precision dairying. Skin surface temperature is an important indicator for the diagnosis of cow's illnesses and for the estimation of their physiological status. Infrared thermography (IRT) is a simple, effective, on-site, and noninvasive method that detects surface heat, which is emitted as infrared radiation and generates pictorial images without causing radiation exposure. In human and bovine medicine, IRT is used as a diagnostic tool for assessment of normal and physiological status.

Soft computing modelling of moisture sorption isotherms of milk-foxtail millet powder and determination of thermodynamic properties.

Moisture sorption isotherms of spray-dried milk-foxtail millet powder were determined at 10, 25 and 40 °C. Sorption data was fitted using classical and soft-computing approaches. The isotherms were of type II, and equilibrium moisture content (EMC) was temperature dependent. The BET monolayer moisture content decreased from 3.30 to 2.67 % as temperature increased from 10 to 40 °C. Amongst the classical models, Ferro-Fontan gave the best fit of EMC-aw data. However, the Sugeno-type adaptive neuro-fuzzy inference system (ANFIS) with generalized bell-shaped membership function performed better than artificial neural network and classical models with RMSE as low as 0.0099. The isosteric heat of sorption decreased from 150.32 kJ mol(-1) at 1 % moisture content to 44.11 kJ mol(-1) at 15 % moisture. The enthalpy-entropy compensation theory was validated, and the isokinetic and harmonic mean temperatures were determined as 333.1 and 297.5 K, respectively.

Effect of enzymatic hydrolysis of starch on pasting, rheological and viscoelastic properties of milk-barnyard millet (Echinochloa frumentacea) blends meant for spray drying.

The influence of enzymatic hydrolysis of starch on the pasting properties of barnyard millet was studied using a rheometer. The effects of blending hydrolyzed barnyard millet wort with milk at different ratios (0:1, 1:1, 1:1.5 and 1:2) on flow and viscoelastic behavior were investigated. From the pasting curves, it was evident that enzymatically-hydrolyzed starch did not exhibit typical pasting characteristics expected of normal starch. The Herschel-Bulkley model fitted well to the flow behaviour data, with coefficient of determination (R(2)) ranging from 0.942 to 0.988. All milk-wort blends demonstrated varying degree of shear thinning with flow behavior index (n) ranging from 0.252 to 0.647. Stress-strain data revealed that 1:1 blend of milk to wort had the highest storage modulus (7.09-20.06Pa) and an elastically-dominant behavior (phase angle <45°) over the tested frequency range. The crossover point of G' and G" shifted to higher frequencies with increasing wort content. From the flow and viscoelastic behavior, it was concluded that the 1:1 blend of milk to wort would have least phase separation and better flowability during spray drying.

Investigation of body and udder skin surface temperature differentials as an early indicator of mastitis in Holstein Friesian crossbred cows using digital infrared thermography technique.

AIM: The objective of this study was to investigate the ability of infrared thermography (IRT) technique and its interrelationship with conventional mastitis indicators for the early detection of mastitis in Holstein Friesian (HF) crossbred cows. MATERIALS AND METHODS: A total of 76 quarters of lactating HF crossbred (Bos indicus × Bos taurus) cows (n=19) were monitored for body temperature (i.e., eye temperature) and udder skin surface temperature (USST) before milking using forward-looking infrared (FLIR) i5 camera. Milk samples were collected from each quarter and screened for mastitis using Somatic Cell Count (SCC), Electrical Conductivity (EC), and California mastitis test. Thermographic images were analyzed using FLIR Quick Report 1.2 image analysis software. Data on body and USST were compiled and analyzed statistically using SPSS 16.0 and Sigmaplot 11. RESULTS: The mean±standard deviation (SD) body (37.23±0.08°C) and USST (37.22±0.04°C) of non-mastitic cow did not differ significantly; however, the mean USST of the mastitis-affected quarters were significantly higher than the body temperature and USST of unaffected quarters (p<0.001). The mean±SD USST of the subclinical mastitis (SCM) and clinical mastitis-affected quarters were 38.08±0.17 °C and 38.25±0.33 °C, respectively, which is 0.72 and 1.05 °C higher than the USST temperature of unaffected quarters. The USST was positively correlated with EC (r=0.95) and SCC (r=0.93). The receiver operating characteristic curve analysis revealed a higher sensitivity for USST in early prediction of SCM with a cut-off value of >37.61°C. CONCLUSION: It is concluded that infrared thermal imaging technique could be used as a potential noninvasive, quick cow-side diagnostic technique for screening and early detection of SCM and clinical mastitis in crossbred cows.

Preparation and characterization of milk protein films and their application for packaging of Cheddar cheese.

Casein and whey protein concentrate (WPC) films, plasticized with glycerol and sorbitol independently, were prepared by casting. The film thickness, water vapour and oxygen permeation and tensile and moisture sorption properties of the films were determined. The tensile strength (TS), tensile strain (TE) and elastic modulus (EM) of the films ranged from 0.71 to 4.58 MPa, 19.22 to 66.63 % and 2.05 to 6.93 MPa, respectively. The film properties were influenced by the type of biopolymer (casein and whey protein concentrate), plasticizer and its concentration. Increasing the plasticizer concentration, increased the film thickness, TE and water vapour permeability (WVP), but decreased the TS and EM. As the concentration of plasticizer increased to the highest level, the film thickness increased from 0.168 to 0.305 mm for glycerol-plasticized films and from 0.251 to 0.326 mm for sorbitol-plasticized films. The film thickness increased because the amount of plasticizer in the film network increased and the amount of biopolymer remained same. Casein films showed superior tensile properties as compared to WPC films. The WVP of both casein and WPC films lied between 3.87 and 13.97 g.mm./(m(2).h.kPa). The moisture sorption isotherms of both films were typical of high-protein material, and were adequately described by the GAB model. The oxygen permeability of casein films was relatively lower than that of WPC films, regardless of the plasticizer used. The sensory data revealed that the organoleptic quality of Cheddar cheese was unaffected by milk-protein film packaging.

Biodegradation characteristics of starch-polystyrene loose-fill foams in a composting medium.

The structures and biodegradabilities of loose-fill foams, containing starch and polystyrene at ratios of 70:30 and 80:20, were evaluated using a laboratory composting system. Each formulation was foamed (extrusion expanded) using either 0.2% azodicarbonamide or 0.25% citric acid as the chemical blowing agent. Biodegradability, a measure of the quantity of material mineralized, was expressed as the percentage of CO(2) in the exhaust gas eluted from the individual chambers. The CO(2) generation peaked after about 15 days of composting, and then decreased. The rate and amount of CO(2) eluted depended on the starch content in the foams. Similarly, there were significant differences in the rates and quantities of CO(2) emissions for the foams blown with azodicarbonamide versus citric acid. At the end of the composting tests, the remaining foam material had fibrous and crumbly textures, presumably consisting primarily of polystyrene. FTIR and NMR spectra of the foams, taken after 39days of composting, did not reveal the spectral features of starch, thereby confirming the decomposition of the starch.