Bacillus licheniformis is an important spoilage-associated bacterium in the food and dairy industries owing to its ability to form persistent biofilms and withstand conventional sanitation treatments. Screening of 42 samples collected from dairy-associated and environmental sources resulted in the isolation of six lytic bacteriophages (SAR 01–SAR 06) infecting B. licheniformis. The phages were characterized for their morphology, host range, physicochemical stability, infection kinetics, and biocontrol potential. TEM analysis revealed that all phages belonged to the family Siphoviridae and possessed an icosahedral head of approximately 90–100 nm in diameter and a contractile tail of 120–140 nm in length. Host range analysis demonstrated that phage SAR 02 exhibited the broadest host spectrum, infecting strains belonging to nine Bacillus species, whereas SAR 04 displayed the narrowest host range. Most phages infected different strains of B. licheniformis as well as selected Geobacillus and other Bacillus species, while none infected Escherichia coli or Pseudomonas spp. The phages were highly sensitive to pH 2, alkaline conditions, exposure to 80°C, and nitric acid treatment. Additionally, phages SAR 01, SAR 02, and SAR 03 were inactivated by 0.9% sodium dodecyl sulphate (SDS) and 0.4% sodium hydroxide (NaOH). One-step growth curve analysis revealed latent periods ranging from 20 to 40 min and an average burst size of approximately 100 plaque-forming units per infected cell. The phages effectively inhibited the growth of B. licheniformis in vitro and prevented biofilm formation on food-contact surfaces. These findings highlight the potential of lytic bacteriophages as eco-friendly biocontrol agents for managing B. licheniformis contamination and biofilm-associated spoilage in dairy processing environments.

The aim of the study was to determine curd formation in abomasum of the pre-ruminant calves when fed whole milk fortified with probiotics and kitchen herbs (carom seeds, cinnamon and turmeric) as prophylaxis against calf diarrhoea. Ultrasonographic imaging reveals that these products enhanced clotting process and abomasal emptying. It can be suggested that addition of probiotics and herb mix in whole milk accelerated the process of gastric emptying less than 3 hours (hrs) of feeding.

This study aimed to investigate the protective effect of Limosilactobacillus fermentum NCDC701 against AlCl3 and D-galactose induced Alzheimer rat model. The dose of AlCl3 (50 mg/kg bodyweight) and D-galactose (100 mg/kg bodyweight) for 7 days induced oxidative stress such as amyloid plaque deposition, neuroinflammation, and reduction of neurotransmitters in the brain. In contrast, the supplementation of L. fermentum NCDC701 improved the neurotransmitter levels, antioxidants and anti-inflammatory cytokine IL-10, while decreased the amyloid peptide, pro-inflammatory cytokines, as well as LPS in colon and brain. The recovery in amyloid plaque deposition and tissue injury were also confirmed by the histopathological examinations. The supplementation of L. fermentum NCDC701 also improved the decreased expression levels of intestinal tight junction proteins, inhibited the up-regulation of p65, COX-2 and iNOS expression, and improved gut microbiota dysbiosis by increasing the Firmicutes: Bacteroidetes ratio. It also supported beneficial microflora (i.e. Oscillospira, Ruminococcus, Verrucomicrobia, Lactobacillus, and Prevotella), and suppressed the pathogenic microflora (i.e. Spirochaetes, Tenericutes, Prophyromonadaceae, Coprococcus, Clostridium, and Allobaculum) by increasing the concentration of butyrate and total short-chain fatty acids compared to the AlCl3 and D-galactose treated model. This finding provides insights into the effect of L. fermentum NCDC701 intervention on the gut–microbiome–brain axis and should assist future understanding of probiotics for improved host health.

Comprehensive safety assessment of potential probiotic strains is crucial in the selection of risk-free strains for clinical translation. This study aimed to evaluate the biosafety of Limosilactobacillus fermentum NCDC 400, a potential probiotic strain, using oral toxicity tests in a Swiss albino mouse model. Mice were orally gavaged with low (108 CFU/mouse/day) and high (1010 CFU/mouse/day) doses of NCDC 400 for 14 (acute), 28 (subacute), and 90 (subchronic) days to assess behavioral, hematological, biochemical, immunological, and histological effects. The administration of NCDC 400 did not result in any observable adverse effects on general health parameters, including body weight, feed and water intake, and organ indices. Hematological and biochemical parameters, such as glucose, serum enzymes, urea, creatinine, serum minerals, total serum proteins, and lipid profile, remained largely unaffected by the test strain. Notably, NCDC 400 administration led to a significant reduction in harmful intestinal enzymes and improvement in gut health indices, as indicated by fecal pH, lactate, ammonia, and short-chain fatty acids. There were no instances of bacterial translocation of NCDC 400 to blood or extra-intestinal organs. Immune homeostasis was not adversely affected by repeated exposure to NCDC 400 in all three oral toxicity studies. Histopathological examination revealed no strain-related changes in various tissues. Based on these findings, a dose of 1010 CFU/mouse/day was considered as the No Observable Effect Level (NOEL) in healthy mice. In conclusion, this study demonstrates the safe and non-toxic behavior of L. fermentum NCDC 400. The results support and ensure the safety and suitability for clinical trials and eventual translation into clinical practice as potential probiotic.

The present study was aimed to assess and validate the safety and functional efficacy of an indigenous probiotic strain Limosilactobacillus fermentum NCDC 400 (hereafter, LFN400) in an immunocompromised murine model. The study included four groups; a normal control (NC) group without immune suppression; an experimental model control (MC) with immune suppression induced via intraperitoneal cyclophosphamide (Cy) administration; and two MC groups orally administered with either low dose (LD) or high dose (HD) of LFN400 at dose 108 and 1010 CFU/mouse/day, respectively, for 15-days. Both control groups received normal saline as placebo control. LFN400 improved specific experimental characteristics including hematological and serum biochemical markers. Compared to MC group, LFN400-fed groups showed markedly (P < 0.05) decreased arrays of detrimental caecal enzymes. We did not observe instances of bacterial translocation of LFN400 from gut to bloodstream or extra-intestinal organs. LFN400 intake significantly (P < 0.05) enhanced spleen cell differentiation, immune and oxidative stress markers, and restored Cy-induced histopathological changes in multiple tissues, including the spleen. There was no genotoxic effect of LFN400 on bone marrow cells. Although not statistically significant, LFN400 feeding moderately increased gut microbiome diversity, supporting the growth of beneficial saccharolytic microorganisms and reducing the presence of pathobionts. The findings demonstrate that the probiotic strain LFN400 possesses in vivo safety and immunomodulatory potency and thus should be considered a potential candidate for future human clinical studies.

Probiotics are amply studied and applied dietary supplements of greater consumer acceptance. Nevertheless, the emerging evidence on probiotics-mediated potential risks, especially among immunocompromised individuals, necessitates careful and in-depth safety studies. The traditional probiotic safety evaluation methods investigate targeted phenotypic traits, such as virulence factors and antibiotic resistance. However, the rapid innovation in omics technologies has offered an impactful means to ultimately sequence and unknot safety-related genes or their gene products at preliminary levels. Further validating the genome features using an array of phenotypic tests would provide an absolute realization of gene expression dynamics. For safety studies in animal models, the in vivo toxicity evaluation guidelines of chemicals proposed by the Organization for Economic Co-operation and Development (OECD) have been meticulously adopted in probiotic research. Future research should also focus on coupling genome-scale safety analysis and establishing a link to its transcriptome, proteome, or metabolome for a fine selection of safe probiotic strains. Considering the studies published over the years, it can be inferred that the safety of probiotics is strain-host-dose-specific. Taken together, an amalgamation of in silico, in vitro, and in vivo approaches are necessary for a fine scale selection of risk-free probiotic strain for use in human applications.

We investigated the antioxidant and anti-inflammatory properties of Lacticaseibacillus rhamnosus Ram12-derived EPSRam12 in a D-galactose-induced liver injury mouse model. Initially, EPSRam12 was characterized for its composition, molecular weight, and structural features. It was then administered orally to D-galactose-induced mice (which had received an intraperitoneal injection of D-galactose, 100 mg/kg body weight) at doses of 25 mg/kg (low dose) and 50 mg/kg (high dose) for 45 days. After treatment, biochemical markers, antioxidant status, cytokine levels, and liver inflammatory gene expression were evaluated. The results showed that EPSRam12 was a branched chain heteropolysaccharide comprising mannose, rhamnose, and arabinose monosaccharides with molecular weight of 2.6 million Daltons. EPSRam12, with its unique structural features such as hydroxyl and methyl groups, glycosidic bonds, and functional groups like carboxylates and sulfates, demonstrated promising bioactive properties. Administering EPSRam12 to D-galactose-induced mice resulted in a significant increase in antioxidant enzyme activity and a reduction in oxidative stress indicators. Additionally, it exhibited anti-inflammatory effects by modulating cytokine levels, lowering pro-inflammatory markers, and inhibiting key inflammatory pathways in the liver in a dose-dependent manner. Our findings underscore the potential of EPSRam12 as an effective antioxidant and anti-inflammatory agent, with promising applications in functional foods and pharmaceuticals.

Ensiling straw for livestock feeding is an emerging technique for addressing the fodder shortage in the country, as it optimizes nutrient storage and utilization. In this study, rice straw was mixed with pea waste (pods) and brewer's grains in the ratio of 35:50:15 respectively, and ensiled in triplicate, with mini-silos opened and sampled at intervals of 30, 40, and 50 days.. These ingredients were mixed with exogenous fibrolytic enzyme (EFE, xylanase; X derived from Trichoderma citrinoviride, 1500 IU/kg), coupled with either a solitary homofermentative inoculant (Pediococcus acidilactici NCDC 609; PA), heterofermentative inoculant (Leuconostoc mesenteroides NCDC 421; LM), or a combination of both. Ensiling of peapods, brewer's grain, and additives reduced (p<0.05) pH from 4.4 to 3.9 (control vs combination of xylanase with homo and heterofermentative bacteria (X+PA+LM group), ammonia-N from 0.20 to 0.18% of DM (control vs X+PA+LM group), butyric acid (0.0114 to 0.0110% of DM), and fiber content from 39.91 to 37.20% of DM (control vs X+PA+LM group) in the treatment groups in which ingredients were supplemented with a combination of homofermentative and heterofermentative bacteria with the exogenous enzyme at all time intervals, i.e., 30th, 40th, and 50th days. However, the best quality silage in terms of pH, lactic acid content, and NH3-N was formed after 50 days of fermentation. The addition of enzymes solubilized the fiber-bound nitrogen fraction of ingredients and made it available to LAB, hence enhancing (p<0.05) its crude protein content. Therefore, it is advisable to ensile the rice straw with peapods and brewer's grain in the suggested ratio to meet the feed demands of livestock and address environmental issues.

Aim of present study was to develop an innovative Cleaning-in-place (CIP) system, having control system with instrumentation, for dairy process equipments. Its performance was evaluated for three-stage thin film scraped surface heat exchanger (TS-TFSSHE). CIP system consists of multi-partition tank, control system with instrumentation and fluid flow system. Experiments were conducted with three independent variables (scraper speed: 300, 225 and 150 RPM; solution temperature: 80, 70 and 60 degrees C; solution concentration: 2%, 1.375% and 0.75%) using response surface methodology. CIP performance responses were chemical rinsing duration, total CIP time, total plate count and coliform count. Optimized solution was obtained as 0.85% sodium-hydroxide concentration, 72.7 degrees C CIP solution temperature and 150 RPM scraper speed. Validation of optimized solution showed that predicted response values were comparable with mean experimental values and found non-significant (p>0.05). This CIP system is movable (wheel mounted) and may be used for cleaning of other dairy process equipments at other places as well.

Bacteriophages infect and replicate inside a bacterial host as well as serve as natural bio-control agents. Phages were once viewed as nuisances that caused fermentation failures with cheese-making and other industrial processes, which lead to economic losses, but phages are now increasingly being observed as being promising antimicrobials that can fight against spoilage and pathogenic bacteria. Pathogen-free meals that fulfil industry requirements without synthetic additives are always in demand in the food sector. This study introduces the readers to the history, sources, and biology of bacteriophages, which include their host ranges, absorption mechanisms, lytic profiles, lysogenic profiles, and the influence of external factors on the growth of phages. Phages and their derivatives have emerged as antimicrobial agents, biodetectors, and biofilm controllers, which have been comprehensively discussed in addition to their potential applications in the food and gastrointestinal tract, and they are a feasible and safe option for preventing, treating, and/or eradicating contaminants in various foods and food processing environments. Furthermore, phages and phage-derived lytic proteins can be considered potential antimicrobials in the traditional farm-to-fork context, which include phage-based mixtures and commercially available phage products. This paper concludes with some potential safety concerns that need to be addressed to enable bacteriophage use efficiently.

Dietary patterns play an important role in regards to the modulation and control of the gut microbiome composition and function. The interaction between diet and microbiota plays an important role in order to maintain intestinal homeostasis, which ultimately affect the host's health. Diet directly impacts the microbes that inhabit the gastrointestinal tract (GIT), which then contributes to the production of secondary metabolites, such as short-chain fatty acids, neurotransmitters, and antimicrobial peptides. Dietary consumption with genetically modified probiotics can be the best vaccine delivery vector and protect cells from various illnesses. A holistic approach to disease prevention, treatment, and management takes these intrinsically linked diet-microbes, microbe-microbe interactions, and microbe-host interactions into account. Dietary components, such as fiber can modulate beneficial gut microbiota, and they have resulting ameliorative effects against metabolic disorders. Medical interventions, such as antibiotic drugs can conversely have detrimental effects on gut microbiota by disputing the balance between Bacteroides and firmicute, which contribute to continuing disease states. We summarize the known effects of various dietary components, such as fibers, carbohydrates, fatty acids, vitamins, minerals, proteins, phenolic acids, and antibiotics on the composition of the gut microbiota in this article in addition to the beneficial effect of genetically modified probiotics and consequentially their role in regards to shaping human health. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Misti dahi, a popular fermented milk product in eastern India, is experiencing growing demand across the country. To meet the increasing demand, it is essential to identify sugar-tolerant starter cultures that can ensure more desirable and consistent fermentation outcomes while minimizing the risk offer mentation failure. This study aimed to isolate, screen, and identify sugar-tolerant cultures suitable for Misti dahi production. Out of ten isolated cultures, S. thermophilus MD3 exhibited remarkable tolerance to high sugar concentrations of up to 20%in milk without compromising cell viability and curdling time. Moreover, it consistently produced Misti dahi of superior quality in terms of physicochemical, microbiological, textural, sensory, and rheological attributes. The isolated S. thermophilus MD3culture holds potential for commercialization in Misti dahi production

Fructophilic Lactic Acid Bacteria (FLAB), Fructobacillus fructosus DPC7238 and pseudofructophilic Leuconostoc mesenteroides DPC7261 and non-FLAB Limosilactobacillus reuteri DSM20016 strains were studied for their growth and morphological evolution as a function of increased fructose concentrations (0, 25, and 50% w/v) in the media. A comparison of the genomics of these strains was carried out to relate observed changes and understand fructose-rich adaptations. The viability of FLAB strains were reduced by approx. 50% at a 50% fructose concentration, while the Limosilactobacillus reuteri strain was reduced to approx. 98%. Electron microscopy demonstrated that FLAB strain, Fructobacillus. fructosus and pseudofructophilic Leuc. mesenteroides, were intact but expanded in the presence of high fructose in the medium. Limosilactobacillus reuteri, on the other hand, ruptured as a result of excessive elongation, resulting in the formation of cell debris when the medium contained more than 25% (w/v) fructose. This was entirely and quantitatively corroborated by three-dimensional data obtained by scanning several single cells using an atomic force microscope. The damage caused the bacterial envelope to elongate lengthwise, thus increasing width size and lower height. The cell surface became comparatively smoother at 25% fructose while rougher at 50% fructose, irrespective of the strains. Although Fructobacillus fructosus was highly fructose tolerant and maintained topological integrity, it had a comparatively smaller genome than pseudofructophilic Leuc. mesenteroides. Further, COG analysis identified lower but effective numbers of genes in fructose metabolism and transport of Fructobacillus fructosus, essentially needed for adaptability in fructose-rich niches.

The safety of probiotic bacterial strains has been the subject of considerable discussion due to emerging health risks associated with their use in various health conditions. Herein, we integrate and execute both in silico and in vitro approaches to evaluate the safety of an indigenous probiotic strain, Limosilactobacillus fermentum NCDC 400. Our analysis of the strain's whole genome sequencing using established pipelines and universally accepted databases find no potential antibiotic-resistance or virulence genes of public health concern. Functional annotation of the genome indicates that NCDC 400 does not produce unusual metabolites or harmful enzymes. In vitro testing confirms that the strain does not exhibit any harmful phenotypes, activities of harmful enzymes, or overproduction of harmful metabolites. The strain is sensitive to the antagonistic properties of human serum as well as to all antibiotics enlisted by EFSA guidelines. Targeted PCR assays confirm the absence of genes conferring unusual antibiotic resistance and virulence factors. Cell culture assays reveal that NCDC 400 is non-cytotoxic to Caco-2 cells and does not affect their cellular integrity. Overall, these results demonstrate the safe and non-toxic nature of NCDC 400 and call for further in vivo trials to evaluate and establish its safety in healthy and diseased preclinical models.

The present study compared pre-treatment effect of exogenous fibrolytic enzyme (EFE) and lactic acid bacteria (LAB) inoculants on the quality of sugarcane tops silage (SCT). Sugarcane tops (301 g DM/kg fresh matter) were chopped into 2.5–3.0 cm lengths and sub-sampled in 33 (11 × 3; treatments × replications) batches. A total of eleven treatments, including two controls, i.e. C1 (without additives) and C2 (with common additives: 0.5% NaCl, 0.5% urea and 2% molasses as such) and nine treatments were prepared in factorial arrangements of (3 × 3) exogenous fibrolytic enzyme (C cellulose, X xylanase and C + X) with LAB inoculants (LF Lactobacillus fermentum, PA Pediococcous acidilactici and LF + PA). After 30 days of ensiling samples were analysed; pH, lignocelluloses, butyric acid, oxalate, yeast and mould count were reduced (p < 0.05) in all treatments, whereas, the lactic acid (LA), acetic acid (AA) and metabolisable energy (ME) content were increased (p < 0.05) in all treatments than controls, C1 and C2 SCT silage. DM loss and NH3–N (% total nitrogen) was found higher (p < 0.05) in C2 and all treatments as compared to the C1 silage. The effect of EFE and LAB interaction was found significant (p < 0.01) for LA:AA, Fleig point and butyric acid content of SCT silage. The efficacy of EFE and LAB inoculant was higher (p < 0.05) when used in combinations, cellulase + xylanase and LF + PA as compared to either type. Silage Fleig point value was found highest (p < 0.05) in LF + PA + X and LF + PA + C + X treatments. Overall comparative effectiveness suggested that xylanase with Lactobacillus fermentum plus Pediococcous acidilactici were the most promising combinations to improve sugarcane tops silage quality.

Exopolysaccharides (EPS) are acknowledged for their diverse functional and technological properties. This study presents the characterization of EPS400, an acidic exopolysaccharide sourced from the native probiotic Limosilactobacillus fermentum NCDC400. Notably, this strain has demonstrated previous capabilities in enhancing dairy food texture and displaying in vivo hypocholesterolemic activity. Our investigation aimed to unveil EPS400′s potential biological roles, encompassing antioxidant, antibacterial, and immunomodulatory activities. The results underscore EPS400′s prowess in scavenging radicals, including the 2,2-diphenyl-1-picrylhydrazyl radical, 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) radical, superoxide radical, hydroxyl radical, and chelating activity targeting the ferrous ion. Furthermore, EPS400 displayed substantial antibacterial effectiveness against prevalent food spoilage bacteria such as Pseudomonas aeruginosa NCDC105 and Micrococcus luteus. Remarkably, EPS400 exhibited the ability to modulate cytokine production, downregulating pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and nitric oxide, while concurrently promoting the release of anti-inflammatory cytokine IL-10 within lipopolysaccharide-activated murine primary macrophages. Additionally, EPS400 significantly (p ≤ 0.05) enhanced the phagocytic potential of macrophages. Collectively, our findings spotlight EPS400 as a promising contender endowed with significant antioxidant, antibacterial, and immunomodulatory attributes. These characteristics propose EPS400 as a potential pharmaceutical or bioactive component, with potential applications in the realm of functional food development.

Lactic acid bacteria (LAB) are the consortium of important microorganisms that belong to Gram-positive, catalase-negative, non-spore-forming bacteria with cocci or rod-shaped morphology. These are generally used as starter cultures in food and dairy fermentations. Although ubiquitous in nature, their abundance is naturally higher in the lactose-rich niches, viz., milk and other fermented milk products. Besides producing lactic acid from lactose, they produce a wide array of metabolites like carbon dioxide, bacteriocin, hydrogen peroxide, diacetyl, acetaldehyde, etc., which not only enjoy sensory appeal but also extend shelf stability of the food products by assuring their safety. Additionally, several of these LAB demonstrate numerous health benefits on the host in a strain-host-dependent manner and are termed "probiotics." Hence, the isolation of pure lactic cultures and their characterization either as starter cultures for dairy fermentations or as probiotics for human health are of great economic and techno-functional significance. This chapter highlights the scheme of isolation of LAB and bacteriocinogenic dairy starters as pure cultures from various eco-niche and their subsequent phenotypic and genotypic characterization methods. Additionally, this chapter also ponders 256on some crucial in vitro tests that are vital in ascertaining the probiotic and safety potential of LAB.

The prevalence of iron deficiency anaemia is a significant issue worldwide, affecting individuals of all ages and often associated with inadequate iron bioavailability. Despite the use of ferrous salt supplements to address anaemia, their limited bioaccessibility and bioavailability in human GIT and adverse impact on food properties remain significant challenges. Hence, this study aims to explore the iron chelation mechanism of an exopolysaccharide EPSKar1 to enhance iron bioaccessibility, bioavailability, and anti-anaemic effects using cell culture and an anaemic rat model. EPSKar1 was extracted from Lacticaseibacillus rhamnosus Kar1 and complexed with FeSO4 to form "EPSKar1-iron". This novel complex, besides being bio-accessible after in vitro gastric digestion, demonstrated 61.27 ± 1.96% iron bioavailability to the Caco-2 cells. In line with these in vitro findings, intragastric administration of the EPSKar1-iron complex to anaemic Wistar rats at 25 and 50 mg per kg body weight significantly restored blood haemoglobin levels and re-established the morphological features of red blood cells. Furthermore, the apparent digestibility co-efficient and iron uptake improved significantly without adversely affecting the serum biochemical parameters in these anaemic rats. The levels of iron-transport proteins including serum transferrin and ferritin in tissue and plasma have increased remarkably upon oral administration of EPSKar1-iron at a higher dose of 50 mg per kg body weight. Oral supplementation of EPSKar1-iron did not foster adverse histological changes in the liver, kidneys, and spleen. In fact, the treatment with the EPSKar1-iron complex had a restitution effect on the tissue architecture, thereby ameliorating the tissue lesions. These findings collectively indicate that the EPSKar1-iron complex shows nutraceutical potential in enhancing the bioavailability of iron and could be a promising approach to tackle iron deficiency anaemia.

Iron is a micronutrient essential for human health and physiology. Iron-deficiency anemia, the most common form of anemia, may occur from an iron homeostasis imbalance. Iron fortification is a promising and most sustainable and affordable solution to tackle the global prevalence of this anemia. Herein, we investigate physicochemical, rheological and stability characteristics of a novel exopolysaccharide ‘EPSKar1’ (derived from Lacticaseibacillus rhamnosus strain Kar1) and its iron complex ‘EPSKar1-Fe (II)’. Our findings demonstrate that EPSKar1 is a high molecular-weight (7.8 × 105 Da) branched-chain heteropolysaccharide composed of galactose, N-acetylglucosamine, and mannose in a molar ratio of 8:4:1, respectively, and exhibits strong emulsifying and water-holding capacities. We find that EPSKar1 forms strong complexes with Fe, wherein the interactions between EPSKar1-Fe (II) complexes are mediated by sulfate, carboxyl, and hydroxyl groups. The rheological analyses reveal that the EPSKar1 and EPSKar1-Fe (II) complexes exhibited shear thickening and thinning properties in skim milk and water, respectively; however, the suspension of EPSKar1 in skim milk is viscoelastic with predominantly elastic response (G'>G" and tan δ < 1). In comparison, EPSKar1-Fe (II) complex exhibits remarkable stability under various processing conditions, highlighting its usefulness for the development of fortified dairy products. Together, these findings underpin considerable prospects of EPSKar1-Fe (II) complex as a novel iron-fortifier possessing multifarious rheological benefits for food applications.