Zinc Oxide Nanoparticles Modified with Galactose as Potential Drug Carrier with Reduced Releasing of Zinc Ions

The toxicity of bare zinc oxide nanoparticles used as drug carriers may be the result of releasing zinc ions. Thus, zinc oxide nanoparticles modified with galactose were obtained. The process of their formation was conducted in the microwave field. The physicochemical properties of the obtained products were studied. The size and electrokinetic potential were defined by using dynamic light scattering technique. The crystalline properties were assessed by X-ray diffractometry. In order to confirm the formation of the desired products, Fourier-transform infrared spectroscopy was used. Releasing of zinc ions from the prepared products when comparing to the bare oxide was analyzed. It was found out that modification of zinc oxide nanoparticles with galactose limits the releasing of zinc ions which are responsible for the toxic effect of the whole carrier-drug conjugate.

Chemistry and Biological Activity of Feed Additive for Poultry Farming

Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Formulation and Technology of the Composition of Essential Oils as a Feed Additive in Poultry with Antibacterial Action

This paper focuses on the formulation of phytobiotic designated for further implantation in poultry farming. Composition was meant to be water-soluble powder containing antibacterial essential oils. The development process involved Thyme, Monarda and Clary sage essential oils. The antimicrobial activity of essential oils composite was meant to be tested against gram-negative and gram-positive bacterial strains. The results are processed using the statistical program Sigma STAT. To make essential oils composition water soluble surfactants were added to them. At the first stage of the study, nine options for the optimal composition of essential oils and surfactants were developed. The effect of the amount of surfactants on the essential oils composition solubility in water has been investigated. On the basis of biopharmaceutical studies, the formulation of phytobiotic has been determined: Thyme, monarda and clary sage essential oils 2:1:1 - 100 parts; Licorice extract 5.25 parts and inhalation lactose 300 parts. A technology for the preparation of phytobiotic has been developed and a technological scheme for the preparation of phytobiotic has been made up. The research was performed within the framework of the grant project CARYS-19-363 funded be the Shota Rustaveli National Science Foundation of Georgia.

Effect of Different Salts on Pseudomonas taetrolens’ Ability to Lactobionic Acid Production

Lactobionic acid is a disaccharide formed from gluconic acid and galactose, and produced by oxidation of lactose. Productivity of lactobionic acid by microbial synthesis can be affected by various factors, and one of them is a presence of potassium, magnesium and manganese ions. In order to extend lactobionic acid production efficiency, it is necessary to increase the yield of lactobionic acid by optimising the fermentation conditions and available substrates for Pseudomonas taetrolens growth. The object of the research was to determinate the application of K2HPO4, MnSO4, MgSO4 × 7H2O salts in different concentration for effective lactose oxidation to lactobionic acid by Pseudomonas taetrolens. Pseudomonas taetrolens NCIB 9396 (NCTC, England) and Pseudomonas taetrolens DSM 21104 (DSMZ, Germany) were used for the study. The acid whey was used as the study object. The content of lactose in whey samples was determined using MilcoScanTM Mars (Foss, Denmark) and high performance liquid chromatography (Shimadzu LC 20 Prominence, Japan). The content of lactobionic acid in whey samples was determined using the high performance liquid chromatography. The impact of studied salts differs, Mn2+ and Mg2+ ions enhanced fermentation instead of K+ ions. Results approved that Mn2+ and Mg2+ ions are necessary for Pseudomonas taetrolens growth. The study results will help to improve the effectiveness of lactobionic acid production with Pseudomonas taetrolens NCIB 9396 and DSM 21104.

Recent Advances in the Valorization of Goat Milk: Nutritional Properties and Production Sustainability

Goat dairy products are gaining popularity worldwide. In developing countries, but also in many marginal regions of the Mediterranean area, goats represent a great part of the economy and ensure food security. In fact, these small ruminants are able to convert efficiently poor weedy plants and small trees into traditional products of high nutritional quality, showing great resilience to different climatic and environmental conditions. In developed countries, goat milk is appreciated for the presence of health-promoting compounds, bioactive compounds such as conjugated linoleic acids, oligosaccharides, sphingolipids and polyammines. This paper focuses on the recent advances in literature on the nutritional properties of goat milk and on innovative techniques to improve its quality as to become a promising functional food. The environmental sustainability of different methodologies of production has also been examined. Goat milk is valued today as a food of high nutritional value and functional properties as well as small environmental footprint. It is widely consumed in many countries due to high nutritional value, lower allergenic potential, and better digestibility when compared to bovine milk, that makes this product suitable for infants, elderly or sensitive patients. The main differences in chemical composition between a cow and goat milk rely on fat globules that in goat milk are smaller and in fatty acids that present a smaller chain length, while protein, fat, and lactose concentration are comparable. Milk nutritional properties have demonstrated to be strongly influenced by animal diet, genotype, and welfare, but also by season and production systems. Furthermore, there is a growing interest in the dairy industry in goat milk for its relatively high concentration of prebiotics and a good amount of probiotics, which have recently gained importance for their therapeutic potential. Therefore, goat milk is studied as a promising matrix to develop innovative functional foods. In addition to the economic and nutritional value, goat milk is considered a sustainable product for its small environmental footprint, as they require relatively little water and land, and less medical treatments, compared to cow, these characteristics make its production naturally vocated to organic farming. Organic goat milk production has becoming more and more interesting both for farmers and consumers as it can answer to several concerns like environment protection, animal welfare and economical sustainment of rural populations living in marginal lands. These evidences make goat milk an ancient food with novel properties and advantages to be valorized and exploited.

Formulation and Evaluation of Dispersible Tablet of Furosemide for Pediatric Use

The objective of this work is to formulate a dry dispersible form of furosemide in the context of pediatric dose adjustment. To achieve this, we have produced a set of formulas that will be tested in process and after compression. The formula with the best results will be improved to optimize the final shape of the product. Furosemide is the most widely used pediatric diuretic because of its low toxicity. The manufacturing process was chosen taking into account all the data relating to the active ingredient and the excipients used and complying with the specifications and requirements of dispersible tablets. The process used to prepare these tablets was wet granulation. Different excipients were used: lactose, maize starch, magnesium stearate and two superdisintegrants. The mode of incorporation of super-disintegrant changes with each formula. The use of super-disintegrant in the formula allowed optimization of the disintegration time. Prepared tablets were evaluated for weight, content uniformity, hardness, disintegration time, friability and in vitro dissolution test. 

Ultrasound Assisted Cooling Crystallization of Lactose Monohydrate

α-lactose monohydrate is widely used in the pharmaceutical industries as an inactive substance that acts as a vehicle or a medium for a drug or other active substance. It is a byproduct of dairy industries, and the recovery of lactose from whey not only boosts the improvement of the economics of whey utilization but also causes a reduction in pollution as lactose recovery can reduce the BOD of whey by more than 80%. In the present study, levels of process parameters were kept as initial lactose concentration (30-50% w/w), sonication amplitude (20-40%), sonication time (2-6 hours), and crystallization temperature (10-20 oC) for the recovery of lactose in ultrasound assisted cooling crystallization. In comparison with cooling crystallization, the use of ultrasound enhanced the lactose recovery by 39.17% (w/w). The parameters were optimized for the lactose recovery using Taguchi Method. The optimum conditions found were initial lactose concentration at level 3 (50% w/w), amplitude of sonication at level 2 (40%), the sonication time at level 3 (6 hours), and crystallization temperature at level 1 (10 °C). The maximum recovery was found to be 85.85% at the optimum conditions. Sonication time and the initial lactose concentration were found to be significant parameters for the lactose recovery.

Nutritional Potential and Functionality of Whey Powder Influenced by Different Processing Temperature and Storage

Whey is an excellent food ingredient owing to its high nutritive value and its functional properties. However, composition of whey varies depending on composition of milk, processing conditions, processing method, and its whey protein content. The aim of this study was to prepare a whey powder from raw whey and to determine the influence of different processing temperatures (160 and 180 °C) on the physicochemical, functional properties during storage of 180 days and on whey protein denaturation. Results have shown that temperature significantly (P < 0.05) affects the pH, acidity, non-protein nitrogen (NPN), protein total soluble solids, fat and lactose contents. Significantly (p < 0.05) higher foaming capacity (FC), foam stability (FS), whey protein nitrogen index (WPNI), and a lower turbidity and solubility index (SI) were observed in whey powder processed at 160 °C compared to whey powder processed at 180 °C. During storage of 180 days, slow but progressive changes were noticed on the physicochemical and functional properties of whey powder. Reverse phase-HPLC analysis revealed a significant (P < 0.05) effect of temperature on whey protein contents. Denaturation of β-Lactoglobulin is followed by α-lacalbumin, casein glycomacropeptide (CMP/GMP), and bovine serum albumin (BSA).

Studying the Effect of Ethanol and Operating Temperature on Purification of Lactulose Syrup Containing Lactose

Lactulose is a synthetic disaccharide which has remarkable applications in food and pharmaceutical fields. Lactulose is not found in nature and it is produced by isomerization reaction of lactose in an alkaline environment. It should be noted that this reaction has a very low yield since significant amount of lactose stays un-reacted in the system. Basically, purification of lactulose is difficult and costly. Previous studies have revealed that solubility of lactose and lactulose are significantly different in ethanol. Considering the fact that solubility is also affected by temperature itself, we investigated the effect of ethanol and temperature on separation process of lactose from the syrup containing lactose and lactulose. For this purpose, a saturated solution containing lactulose and lactose was made at three different temperatures; 25⁰C (room temperature), 31⁰C, and 37⁰C first.  Five samples containing 2g saturated solution was taken and then 2g, 3g, 4g, 5g, and 6g ethanol separately was added to the sampling tubes. Sampling tubes were kept at respective temperatures afterward. The concentration of lactose and lactulose after separation process measured and analyzed by High Performance Liquid Chromatography (HPLC). Results showed that ethanol has such a greater impact than operating temperature on purification process. Also, it was observed that the maximum rate of separation occurred at initial amount of added ethanol.

Isolation and Screening of Fungal Strains for β-Galactosidase Production

Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Untargeted Small Metabolite Identification from Thermally Treated Tualang Honey

This study investigated the effects of thermal treatment on Tualang honey sample in terms of honey colour and heat-induced small metabolites. The heating process was carried out in a temperature controlled water batch at 90oC for 4 hours. The honey samples were put in cylinder tubes with the dimension of 1 cm diameter and 10 cm length for homogenous heat transfer. The results found that the thermal treatment produced not only hydroxylmethylfurfural, but also other harmful substances such as phthalic anhydride and radiolytic byproducts. The degradation of honey protein was due to the detection of free amino acids such as cysteine and phenylalanine in heat-treated honey samples. Sugar dehydration was also occurred because fragmented di-galactose was identified based on the presence of characteristic ions in the mass fragmentation pattern. The honey colour was found getting darker as the heating duration was increased up to 4 hours. Approximately, 60 mm PFund of increment was noticed for the honey colour with the colour change rate of 14.8 mm PFund per hour. Based on the principal component analysis, the score plot clearly shows that the chemical profile of Tualang honey was significantly altered after 2 hours of heating at 90oC.

Chemical Characterization and Prebiotic Effect of Water-Soluble Polysaccharides from Zizyphus lotus Leaves

In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% is neutral sugar and the rest 16.25±1.62% is uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructooligosaccharide (RP95). The oligosaccharides concentration was 1g/L of Man, Rogosa, Sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Utilization of Whey for the Production of β-Galactosidase Using Yeast and Fungal Culture

Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.

Effect of Humidity on in-Process Crystallization of Lactose during Spray Drying

The effect of various humidities on process yields and degrees of crystallinity for spray-dried powders from spray drying of lactose with humid air in a straight-through system have been studied. It has been suggested by Williams–Landel–Ferry kinetics (WLF) that a higher particle temperature and lower glass-transition temperature would increase the crystallization rate of the particles during the spray-drying process. Freshly humidified air produced by a Buchi-B290 spray dryer as a humidifier attached to the main spray dryer decreased the particle glass-transition temperature (Tg), while allowing the particle temperature (Tp) to reach higher values by using an insulated drying chamber. Differential scanning calorimetry (DSC) and moisture sorption analysis were used to measure the degree of crystallinity for the spray-dried lactose powders. The results showed that higher Tp-Tg, as a result of applying humid air, improved the process yield from 21 ± 4 to 26 ± 2% and crystallinity of the particles by decreasing the latent heat of crystallization from 43 ± 1 to 30 ± 11 J/g and the sorption peak height from 7.3 ± 0.7% to 6 ± 0.7%.

Statistical Modeling for Permeabilization of a Novel Yeast Isolate for β-Galactosidase Activity Using Organic Solvents

The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.

Development and Optimization of Colon Targeted Drug Delivery System of Ayurvedic Churna Formulation Using Eudragit L100 and Ethyl Cellulose as Coating Material

The purpose of this study was to prepare time and pH dependent release tablets of Ayurvedic Churna formulation and evaluate their advantages as colon targeted drug delivery system. The Vidangadi Churna was selected for this study which contains Embelin and Gallic acid. Embelin is used in Helminthiasis as therapeutic agent. Embelin is insoluble in water and unstable in gastric environment so it was formulated in time and pH dependent tablets coated with combination of two polymers Eudragit L100 and ethyl cellulose. The 150mg of core tablet of dried extract and lactose were prepared by wet granulation method. The compression coating was used in the polymer concentration of 150mg for both the layer as upper and lower coating tablet was investigated. The results showed that no release was found in 0.1 N HCl and pH 6.8 phosphate buffers for initial 5 hours and about 98.97% of the drug was released in pH 7.4 phosphate buffer in total 17 Hours. The in vitro release profiles of drug from the formulation could be best expressed first order kinetics as highest linearity (r2= 0.9943). The results of the present study have demonstrated that the time and pH dependent tablets system is a promising vehicle for preventing rapid hydrolysis in gastric environment and improving oral bioavailability of Embelin and Gallic acid for treatment of Helminthiasis.

Characterization of a Novel Galactose-Binding Lectin Homologue from Tenebrio molitor

An expressed sequence tag (EST) analysis provideus portions of expressed genes. We have constructed cDNA library and determined randomly sequences from cDNA library clones of T. molitor injected with acholeplasma lysate. We identified the homologous to a galectin gene. As the result of cloning and characterization of novel, we found that the protein has an open reading frame (ORF) of 495 bp, with 164 amino acid residues and molecular weight of 18.5 kDa. To characterize the role of novel Tm-galectin in immune system, we quantified the mRNA level of galectin at different times after treatment with immune elicitors. The galectin mRNA was up-regulated about 7-folds within 18 hrs. This suggests that Tm-galectin is a novel member of animal lectins, and has a role in the process of pathogen recognition. Our study would be helpful for the study on immune defense system and signaling cascade.

Production of Milk Clotting Protease by Rhizopus Stolonifer through Optimization of Culture Conditions

The present study describes the biosynthesis of a milkclotting protease by solid state fermentation (SSF) of a locally isolated mould, Rhizopus stolonifer. The production medium was prepared using wheat bran at 50% (w/v). The production conditions are optimized by varying 7 parameters: carbon and nitrogen sources, medium moisture, temperature, pH, fermentation time and inoculum-s size. The maximum enzyme synthesis was measured after 96 h of incubation time at temperature of 28°C. The optimum pH determined was 6 and the inoculum size was 3.106spores/ml. The optimum initial moisture content is comprised between 50 to 70%. The formation of milk clotting protease is enhanced when galactose and peptone are used at 10% (w/v) and 1% (w/v) concentrations respectively. The maximum production of milk clotting protease is 120 US/ml.

Long- term Irrigation with Dairy Factory Wastewater Influences Soil Quality

The effects of irrigation with dairy factory wastewater on soil properties were investigated at two sites that had received irrigation for > 60 years. Two adjoining paired sites that had never received DFE were also sampled as well as another seven fields from a wider area around the factory. In comparison with paired sites that had not received effluent, long-term wastewater irrigation resulted in an increase in pH, EC, extractable P, exchangeable Na and K and ESP. These changes were related to the use of phosphoric acid, NaOH and KOH as cleaning agents in the factory. Soil organic C content was unaffected by DFE irrigation but the size (microbial biomass C and N) and activity (basal respiration) of the soil microbial community were increased. These increases were attributed to regular inputs of soluble C (e.g. lactose) present as milk residues in the wastewater. Principal component analysis (PCA) of the soils data from all 11sites confirmed that the main effects of DFE irrigation were an increase in exchangeable Na, extractable P and microbial biomass C, an accumulation of soluble salts and a liming effect. PCA analysis of soil bacterial community structure, using PCR-DGGE of 16S rDNA fragments, generally separated individual sites from one another but did not group them according to irrigation history. Thus, whilst the size and activity of the soil microbial community were increased, the structure and diversity of the bacterial community remained unaffected.

Application of Central Composite Design Based Response Surface Methodology in Parameter Optimization and on Cellulase Production Using Agricultural Waste

Response Surface Methodology (RSM) is a powerful and efficient mathematical approach widely applied in the optimization of cultivation process. Cellulase enzyme production by Trichoderma reesei RutC30 using agricultural waste rice straw and banana fiber as carbon source were investigated. In this work, sequential optimization strategy based statistical design was employed to enhance the production of cellulase enzyme through submerged cultivation. A fractional factorial design (26-2) was applied to elucidate the process parameters that significantly affect cellulase production. Temperature, Substrate concentration, Inducer concentration, pH, inoculum age and agitation speed were identified as important process parameters effecting cellulase enzyme synthesis. The concentration of lignocelluloses and lactose (inducer) in the cultivation medium were found to be most significant factors. The steepest ascent method was used to locate the optimal domain and a Central Composite Design (CCD) was used to estimate the quadratic response surface from which the factor levels for maximum production of cellulase were determined.