H2S-Releasing Versatile Montmorillonite Nanoformulation Trilogically Renovates the Gut Microenvironment for Inflammatory Bowel Disease Modulation.

Abnormal activation of the intestinal mucosal immune system, resulting from damage to the intestinal mucosal barrier and extensive invasion by pathogens, contributes to the pathogenesis of inflammatory bowel disease (IBD). Current first-line treatments for IBD have limited efficacy and significant side effects. An innovative H2S-releasing montmorillonite nanoformulation (DPs@MMT) capable of remodeling intestinal mucosal immune homeostasis, repairing the mucosal barrier, and modulating gut microbiota is developed by electrostatically adsorbing diallyl trisulfide-loaded peptide dendrimer nanogels (DATS@PDNs, abbreviated as DPs) onto the montmorillonite (MMT) surface. Upon rectal administration, DPs@MMT specifically binds to and covers the damaged mucosa, promoting the accumulation and subsequent internalization of DPs by activated immune cells in the IBD site. DPs release H2S intracellularly in response to glutathione, initiating multiple therapeutic effects. In vitro and in vivo studies have shown that DPs@MMT effectively alleviates colitis by eliminating reactive oxygen species (ROS), inhibiting inflammation, repairing the mucosal barrier, and eradicating pathogens. RNA sequencing revealed that DPs@MMT exerts significant immunoregulatory and mucosal barrier repair effects, by activating pathways such as Nrf2/HO-1, PI3K-AKT, and RAS/MAPK/AP-1, and inhibiting the p38/ERK MAPK, p65 NF-κB, and JAK-STAT3 pathways, as well as glycolysis. 16S rRNA sequencing demonstrated that DPs@MMT remodels the gut microbiota by eliminating pathogens and increasing probiotics. This study develops a promising nanoformulation for IBD management.

Exploring the Potential of Montmorillonite as an Antiproliferative Nanoagent against MDA-MB-231 and MCF-7 Human Breast Cancer Cells.

Unlike MCF-7 cells, MDA-MB-231 cells are unresponsive to hormone therapy and often show resistance to chemotherapy and radiotherapy. Here, the antiproliferative effect of biocompatible montmorillonite (Mt) nanosheets on MDA-MB-231 and MCF-7 human breast cancer cells was evaluated by MTT assay, flow cytometry, and qRT-PCR. The results showed that the Mt IC50 for MDA-MB-231 and MCF-7 cells in a fetal bovine serum (FBS)-free medium was ~50 and ~200 µg/mL, and in 10% FBS medium ~400 and ~2000 µg/mL, respectively. Mt caused apoptosis in both cells by regulating related genes including Cas-3, P53, and P62 in MDA-MB-231 cells and Bcl-2, Cas-8, Cas-9, P53, and P62 in MCF-7 cells. Also, Mt arrested MCF-7 cells in the G0/G1 phase by altering Cyclin-D1 and P21 expression, and caused sub-G1 arrest and necrosis in both cells, possibly through damaging the mitochondria. However, fewer gene expression changes and more sub-G1 arrest and necrosis were observed in MDA-MB-231 cells, confirming the higher vulnerability of MDA-MB-231 cells to Mt. Furthermore, MDA-MB-231 cells appeared to be much more vulnerable to Mt compared to other cell types, including normal lung fibroblast (MRC-5), colon cancer (HT-29), and liver cancer (HepG2) cells. The higher vulnerability of MDA-MB-231 cells to Mt was inferred to be due to their higher proliferation rate. Notably, Mt cytotoxicity was highly dependent on both the Mt concentration and serum level, which favors Mt for the local treatment of MDA-MB-231 cells. Based on these results, Mt can be considered as an antiproliferative nanoagent against MDA-MB-231 cells and may be useful in the development of local nanoparticle-based therapies.

pH-responsive bentonite nanoclay carriers control the release of benzothiazolinone to restrain bacterial wilt disease.

Ralstonia solanacearum (R. solanacearum) is one of the most devastating pathogens in terms of losses in agricultural production. Bentonite (Bent) is a promising synergistic agent used in development of effective and environmentally friendly pesticides against plant disease. However, the synergistic mechanism of Bent nanoclays with benzothiazolinone (BIT) against R. solanacearum is unknown. In this work, acid-functionalized porous Bent and cetyltrimethylammonium bromide (CTAB) were employed as the core nanoclays, and BIT was loaded into the clay to form BIT-loaded CT-Bent (BIT@CT-Bent) for the control of bacterial wilt disease. BIT@CT-Bent exhibited pH-responsive release behavior that fit the Fickian diffusion model, rapidly releasing BIT in an acidic environment (pH = 5.5). The antibacterial effect of BIT@CT-Bent was approximately 4 times greater than that of the commercial product BIT, and its biotoxicity was much lower than that of BIT under the same conditions. Interestingly, R. solanacearum attracted BIT@CT-Bent into the nanocomposites and induced cytoplasmic leakage and changes in membrane permeability, indicating an efficient and synergistic bactericidal effect that rapidly reduced bacterial density. In addition, BIT@CT-Bent significantly inhibited R. solanacearum biofilm formation and swimming activity, by suppressing the expression of phcA, solR and vsrC. Indeed, exogenous application of BIT@CT-Bent significantly suppressed the virulence of R. solanacearum on tobacco plants, with control effect of 75.48%, 72.08% and 66.08% at 9, 11 and 13 days after inoculation, respectively. This study highlights the potential of using BIT@CT-Bent as an effective, eco-friendly bactericide to control bacterial wilt diseases and for the development of sustainable crop protection strategies.

Chitosan-based intelligent polymeric networks for site-specific colon medication delivery: A comprehensive study on controlled release of diloxanide furoate and network formation dynamics.

Oral medications are prone to gastric degradation and enzymatic inactivation, diminishing their efficacy. This study investigates a solution by developing intelligent polymeric networks, incorporating chitosan, methacrylic acid, N, N, methylene bisacrylamide, and montmorillonite clay, to enable the controlled release of Diloxanide Furoate (DF), an anti-protozoal drug. Employing a swelling-assisted diffusion technique, drug loading percentages varied from 63.96 % to 76.82 % among different formulations. Increased chitosan and methacrylic acid content enhanced drug loading, while N, N, methylene bisacrylamide and montmorillonite clay demonstrated an inverse relationship affecting diffusion and swelling. Equilibrium swelling studies unveiled formulation-dependent behaviors, with chitosan reducing swelling and methacrylic acid promoting it. Higher N, N, methylene bisacrylamide concentrations decreased swelling, indicating a denser cross-linked structure, while montmorillonite clay reduced hydrophilicity and swelling capacity. Further analyses confirmed successful gel formation, particularly in formulations with higher chitosan, methacrylic acid, and N, N, methylene bisacrylamide content, while montmorillonite clay limited gel fraction due to restricted polymer chain mobility. Techniques such as Fourier transform infrared spectroscopy, Differential scanning calorimetry, and thermal gravimetric analyses supported network development, enhancing thermal stability and cross-linking density. This research underscores the flexibility of polymeric networks for precise drug delivery, offering potential advancements in targeted therapies for various medical conditions.

Evaluation of the Adsorption Efficacy of Bentonite on Aflatoxin M1 Levels in Contaminated Milk.

The existence of aflatoxin M1 (AFM1) in raw milk results in economic losses and public health risks. This research aims to examine the capability of bentonite to adsorb and/or eliminate AFM1 from various raw milk types. In addition, the effects of numerous bentonites (HAFR 1, 2, 3 and 4) on the nutritional characteristics of the milk were studied. Our findings revealed that goat milk had the highest value of AFM1 (490.30 ng/L) in comparison to other milks. AFM1 adsorption was influenced by applying bentonite (0.5 and 1 g) in a concentration-dependent manner for different time intervals (from 0 to 12 h). The percentage of AFM1 reached the maximum adsorption level after 12 h to 100, 98.5 and 98% for bentonites HAFR 3, 1 and 2, respectively. HAFR 3 (1 g bentonite) presented higher adsorption efficiency than other bentonites used in the phosphate buffer saline (PBS) and milk. Residual levels of AFM1 reached their lowest values of 0 and 1.5 ng/L while using HAFR 3 in PBS and milk, respectively. With regard to the influence of bentonite on the nutritional characteristics of milk, there was an increase in fat, protein and solid non-fat ratio while using HAFR 3 and 4, yet decreased lactose in comparison with the control. Scanning Electron Microscopy and Fourier Transform-Infrared Spectroscopy both identified bentonites as superior AFM1 binders. The results demonstrated that bentonite, particularly HAFR 3, was the most effective adsorbent and could thus be a promising candidate for the decontamination of AFM1 in milk.

Potential of montmorillonite modified by an organosulfur surfactant for reducing aflatoxin B1 toxicity and ruminal methanogenesis in vitro.

BACKGROUND: Montmorillonite clay modified by organosulfur surfactants possesses high cation exchange capacity (CEC) and adsorption capacity than their unmodified form (UM), therefore they may elevate the adverse impact of aflatoxin B1 (AFB1) on ruminal fermentation and methanogenesis. Chemical and mechanical modifications were used to innovate the organically modified nano montmorillonite (MNM). The UM was modified using sodium dodecyl sulfate (SDS) and grounded to obtain the nanoscale particle size form. The dose-response effects of the MNM supplementation to a basal diet contaminated or not with AFB1 (20 ppb) were evaluated in vitro using the gas production (GP) system. The following treatments were tested: control (basal diet without supplementations), UM diet [UM supplemented at 5000 mg /kg dry matter (DM)], and MNM diets at low (500 mg/ kg DM) and high doses (1000 mg/ kg DM). RESULTS: Results of the Fourier Transform Infra-Red Spectroscopy analysis showed shifts of bands of the OH-group occurred from lower frequencies to higher frequencies in MNM, also an extra band at the lower frequency range only appeared in MNM compared to UM. Increasing the dose of the MNM resulted in linear and quadratic decreasing effects (P < 0.05) on GP and pH values. Diets supplemented with the low dose of MNM either with or without AFB1 supplementation resulted in lower (P = 0.015) methane (CH4) production, ruminal pH (P = 0.002), and ammonia concentration (P = 0.002) compared to the control with AFB1. Neither the treatments nor the AFB1 addition affected the organic matter or natural detergent fiber degradability. Contamination of AFB1 reduced (P = 0.032) CH4 production, while increased (P < 0.05) the ruminal pH and ammonia concentrations. Quadratic increases (P = 0.012) in total short-chain fatty acids and propionate by MNM supplementations were observed. CONCLUSION: These results highlighted the positive effects of MNM on reducing the adverse effects of AFB1 contaminated diets with a recommended dose of 500 mg/ kg DM under the conditions of this study.

Recombinant Thaumatin-Like Protein (rTLP) and Chitinase (rCHI) from Vitis vinifera as Models for Wine Haze Formation.

Cross-linking net aggregates of thermolabile thaumatin-like proteins (TLPs) and chitinases (CHIs) are the primary source of haze in white wines. Although bentonite fining is still routinely used in winemaking, alternative methods to selectively remove haze proteins without affecting wine organoleptic properties are needed. The availability of pure TLPs and CHIs would facilitate the research for the identification of such technological advances. Therefore, we proposed the usage of recombinant TLP (rTLP) and CHI (rCHI), expressed by Komagataella phaffii, as haze-protein models, since they showed similar characteristics (aggregation potential, melting point, functionality, glycosylation levels and bentonite adsorption) to the native-haze proteins from Vitis vinifera. Hence, rTLP and rCHI can be applied to study haze formation mechanisms on a molecular level and to explore alternative fining methods by screening proteolytic enzymes and ideal adsorptive resins.

Rumen Parameters, Nutrients Digestibility and Milk Production of Lactating Boer Goats Fed Diets Containing Clay Minerals.

<b>Background and Objective:</b> The inclusion of clay minerals in dairy nutrition is getting attention owing to their proven beneficial effects. The current study aimed to evaluate the effect of dietary supplementation of three different clay minerals (bentonite, zeolite and humic acid) on the performance of lactating Boer goats. <b>Materials and Methods:</b> Twenty lactating Boer goats (having an average body weight ~42.7 kg) were divided into four groups (5 animals each) by using a completely randomized design (CRD). Each group was fed with one of four dietary treatments: Control group with basal ration R1: Consisting of concentrate feed mixture (CFM) and clover hay (50:50%, C:R) on a dry matter (DM) basis, R2: Basal ration plus 1% bentonite, R3: Basal ration plus 1.25% zeolite and R4: Basal ration plus 0.5% humic acid. <b>Results:</b> The results revealed that bentonite and humic acid increased (p<0.05) the nutrient digestibility and nutritive values compared to zeolite and control groups. Ruminal ammonia and total volatile fatty acids (TVFAs) contents increased (p<0.05) with supplementation of bentonite. No effect of clays supplementation was observed on plasma total protein, urea and creatinine, however, it increased (p<0.05) the albumin concentration and albumin/globulin ratios compared to the control while decreasing the plasma globulin contents. Supplementation of humic acid increased (p<0.05) the AST concentrations. Bentonite supplementation recorded the highest milk yield (p<0.05) and composition, while the zeolite group had the lowest values. <b>Conclusion:</b> The present study indicated that the inclusion of clay minerals particularly bentonite (at 1%) can positively affect the performance of lactating Boer goats.

Does Bentonite Cause Cytotoxic and Whole-Transcriptomic Adverse Effects in Enterocytes When Used to Reduce Aflatoxin B1 Exposure?

Aflatoxin B1 (AFB1) is a major food safety concern, threatening the health of humans and animals. Bentonite (BEN) is an aluminosilicate clay used as a feed additive to reduce AFB1 presence in contaminated feedstuff. So far, few studies have characterized BEN toxicity and efficacy in vitro. In this study, cytotoxicity (WST-1 test), the effects on cell permeability (trans-epithelial electrical resistance and lucifer yellow dye incorporation), and transcriptional changes (RNA-seq) caused by BEN, AFB1 and their combination (AFB1 + BEN) were investigated in Caco-2 cells. Up to 0.1 mg/mL, BEN did not affect cell viability and permeability, but it reduced AFB1 cytotoxicity; however, at higher concentrations, BEN was cytotoxic. As to RNA-seq, 0.1 mg/mL BEN did not show effects on cell transcriptome, confirming that the interaction between BEN and AFB1 occurs in the medium. Data from AFB1 and AFB1 + BEN suggested AFB1 provoked most of the transcriptional changes, whereas BEN was preventive. The most interesting AFB1-targeted pathways for which BEN was effective were cell integrity, xenobiotic metabolism and transporters, basal metabolism, inflammation and immune response, p53 biological network, apoptosis and carcinogenesis. To our knowledge, this is the first study assessing the in vitro toxicity and whole-transcriptomic effects of BEN, alone or in the presence of AFB1.

Effect of Diet Supplementation with the Mycotoxin Binder Montmorillonite on the Bioavailability of Vitamins in Dairy Cows.

The objective of this study was to determine the effect of the mycotoxin binder montmorillonite (MMT) supplemented in the diet of dairy cows on the bioavailability of vitamins A, D, E, B1 and B6. Six multiparous Holstein-Friesian cows were used in a crossover design with two periods. Treatments were a control diet with or without MMT. Vitamins were infused individually into the abomasum through the ruminal cannula. Blood samples were collected from the jugular vein at 0, 1, 2, 3, 4, 6, 9, 12, 24 and 48 h after the administration of each vitamin. Results showed that vitamin A reached maximal concentration (Tmax) at 5.3 h after dosing, the maximal concentration (Cmax) was 1.2 times higher than the basal concentration (Cbasal), and the area under the curve (AUC) was 739 arbitrary units. Vitamin B6 reached the Tmax at 13 h after dosing, the Cmax was 1.4 times higher than the Cbasal, and the AUC was 222 arbitrary units. No differences were observed in Cbasal, Tmax, Cmax and AUC of vitamin A and B6 between control vs. MMT-supplemented cows. Plasma concentrations of vitamins D, E and B1 had no concentration peaks, and were not affected by MMT addition. The lack of a response suggests that their plasma concentration may be tightly regulated. Results of this study do not show evidence that MMT affects the bioavailability of vitamins A and B6 in vivo.

Bentonite does not affect in vitro ruminal gross fermentations but could modify ruminal metabolome and mineral content. A proof of concept.

Clay minerals, such as bentonite, are used as feed additives capable of adsorbing mycotoxins and heavy metals and have been related to many positive effects on animal health and productivity. However, these compounds seem to induce also side effects and to interact with the intestinal and ruminal microbiota. The present in vitro study is aimed at evaluating the effects of different doses of bentonite on ruminal fermentations, metabolome and mineral content. Five doses of bentonite (0, 2.5, 5, 10 and 50 mg in 150 mL total volume) were incubated (39 °C for 24 h) with a dairy cow Total Mixed Ratio (TMR) and the ruminal fluid obtained from one healthy Holstein lactating cow. The kinetics of gas production (GP) continuously monitored during the incubation evidenced no significant differences in either cumulative GP (mL/g DM) or GP rate (mL/g DM/h) between the treatment groups. After the incubation, metabolome and mineral content of treated ruminal fluids were studied in pooled replicate samples by 1H NMR spectroscopy and Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES), respectively. The NMR analysis led to the identification of 20 metabolites and suggested a clear metabolic differentiation among treatments. The ICP-OES analysis suggested that the addition of bentonite affected the concentration of Al, Ba, Ca, Cr, Mn, Mo and Sr. It is conceivable that bentonite administration does not affect gross ruminal fermentations, while it seems to modify the ruminal metabolome and the concentrations of few minerals in ruminal fluid.

The ameliorative effects of Nigella sativa, thymoquinone, and bentonite against aflatoxicosis in broilers via AFAR and Nrf2 signalling pathways, and down-regulation of caspase-3.

1. Aflatoxins (AFs) are metabolites which especially have toxic effects on proteins, and are detoxified by the aflatoxin-B1 aldehyde reductase (AFAR) pathway. In this pathway, the aldo-keto reductase family 7, member A2 (AKR7A2) enzyme, which is controlled by nucleic-related erythroid factor 2 (Nrf2), plays an active role. However, data on the efficacy of this critical pathway in broilers is limited.2. The aim of the following study was to investigate the changes in the expression levels of AKR7A2, Nrf2, and caspase-3, and the effects of Nigella sativa seeds (NS), thymoquinone (TMQ), and bentonite (BNT) in broilers exposed to AFs.3. One-hundred broilers were divided into ten groups (control (CNT); AF; NS; TMQ; BNT; AF+TMQ; AF+NS; AF+BNT; AF+BNT+NS; AF+BNT+TMQ) and fed for 28 d. AF, TMQ, NS and BNT were added to diets at levels of 2 mg/kg, 300 mg/kg, 50 g/kg and 10 g/kg respectively.4. The addition of AF to the diet decreased AKR7A2 and Nrf2 levels dramatically, but increased caspase-3 (P < 0.01). TMQ, NS and BNT additions to the diet eliminated all negative effects caused by AF (P < 0.01); and AKR7A2 and Nrf2 were further raised in TMQ and NS groups when compared to the control group. TMQ and NS showed a positive effect on detoxification parameters when given together with BNT.5. Supplementation with NS and TMQ enhanced AF detoxification via the AFAR pathway, by increasing AKR7A2 and Nrf2 levels, in addition to reducing hepatocyte apoptosis.

Role of Na-Montmorillonite on Microbially Induced Calcium Carbonate Precipitation.

The use of additives has generated significant attention due to their extensive application in the microbially induced calcium carbonate precipitation (MICP) process. This study aims to discuss the effects of Na-montmorillonite (Na-MMT) on CaCO3 crystallization and sandy soil consolidation through the MICP process. Compared with the traditional MICP method, a larger amount of CaCO3 precipitate was obtained. Moreover, the reaction of Ca2+ ions was accelerated, and bacteria were absorbed by a small amount of Na-MMT. Meanwhile, an increase in the total cementing solution (TCS) was not conducive to the previous reaction. This problem was solved by conducting the reaction with Na-MMT. The polymorphs and morphologies of the CaCO3 precipitates were tested by using X-ray diffraction and scanning electron microscopy. Further, when Na-MMT was used, the morphology of CaCO3 changed from an individual precipitate to agglomerations of the precipitate. Compared to the experiments without Na-MMT in the MICP process, the addition of Na-MMT significantly reduced the hydraulic conductivity (HC) of sandy soil consolidated.

Role of clay in detoxification of aflatoxin B1 in growing Japanese quail with reference to gender.

The present study investigated the influence of the quail diet polluted with aflatoxin B1 (AFB1) and its detoxification by using clay as a feed additive on the growth performance and some blood biochemical components of growing Japanese quail with reference to sex. A total number of 120 Japanese quail chicks (1 week old), was randomly divided into 10 groups (24 chicks/ group). A 5 × 2 factorial arrangement experiment was performed and included five levels of AFB1 (0 ppm, 1 mg/kg AFB1, 1 mg/kg AFB1 + 1% clay, 2 mg/kg AFB1 and 2 mg/kg AFB1 + 1% clay) and two sexes. Birds fed with aflatoxin free diet had significantly (P ≤ 0.05 and 0.01) higher final live body weight, weight gain and lower mortality rate than the other groups. Addition of 1% clay significantly (P ≤ 0.05 and 0.01) improved the growth performance traits and diminished aflatoxin effect when compared to groups without the addition of clay. Obtained results indicated significant (P ≤ 0.05) differences between the two sexes in their response to aflatoxicosis in the final live body weight and weight gain. Our results showed significant (P ≤ 0.01) changes in all blood biochemicals (total protein, albumin, globulin, total cholesterol, creatinine, uric acid) and activities of serum enzymes studied due to the toxicity of AFB1. Conclusively, the consumption of polluted diets with AFB1 caused deleterious effects on the growth performance and blood biochemicals components of Japanese quail, while dietary addition of natural clay to the diet of growing Japanese quail caused beneficial effects.

The Nature of the Increased Efficiency of Sorption-Stimulating Preparations Containing Non-Ionic Surfactants for Pre-Sowing Seed Treatment.

The effect of the introduction of a non-ionogenic surfactant Polysorbate 20 into a sorption preparation (CB-H-BYA) on the structure of sorbent layers formed on the surface of spring wheat seeds during their pre-sowing treatment has been studied using electron microscopy. According to the results, an increase in the efficiency of sorption preparations containing Polysorbate 20 is based on an intensification of the bentonite aggregate disintegration into individual montmorillonite particles and a formation of more dense protective sorption layer providing a better protection of seeds against allelotoxins on the seed surface. The introduction of non-ionogenic surfactants into the preparation increases a sorption capacity of a bentonite-humus complex that results in a decreased gibberellin activity in a solution. Therefore, to achieve the maximum physiological activity of gibberellin in a preparation solution, it is necessary to increase its concentration from 100 to 300 mg/L. As a result, the stimulating effect increases from 36 to 55%.

Calcination Improves the In Vivo Efficacy of a Montmorillonite Clay to Bind Aflatoxin G1 in Broiler Chickens: A Toxicokinetic Approach.

The goal of this study was to investigate the toxicokinetic characteristics of aflatoxin G1 (AFG1) in broiler chickens and the effect of calcination of a Tunisian montmorillonite clay on the in vivo absorption of AFG1. In this study, broiler chickens were randomly distributed into four groups of 10 animals. Group 1 was administered AFG1 (2 mg/kg body weight (BW)) by single intravenous injection (IV), group 2 received an intra-crop bolus (PO) of AFG1 without any clay, group 3 was dosed AFG1 PO together with an oral bolus of purified clay (CP), and group 4 received AFG1 PO with an oral bolus of calcined clay. A significant difference in the area under the curve (AUC0-t) was observed for group 4 (6.78 ± 4.24 h*ng/mL) in comparison with group 2 (12.83 ± 4.19 h*ng/mL). A significant reduction of the oral bioavailability of AFG1 was observed for group 4 (7.61 ± 4.76%) compared with group 2 (14.40 ± 4.70%), while no significant effect was observed of CP. In this experiment, no phase I nor phase II metabolites of AFG1 were observed. These findings confirm that calcination of the purified montmorillonite clay enhances the adsorption of AFG1 in the gastrointestinal tract after oral administration, thereby reducing its bioavailability, thus reducing its toxic effects.

Synergic antimicrobial properties of Carvacrol essential oil and montmorillonite in biodegradable starch films.

This work evaluated the synergic effect of carvacrol essential oil (EO) and montmorillonite (MMT) (named hybrid compound), incorporated in thermoplastic starch (TPS) films in different contents (4.5, 9, and 15 wt%). TPS films were characterized by FTIR, FT-Raman, DSC, XRD, and antimicrobial assay. FTIR and FT-Raman indicated new hydrogen bonds between hybrid and starch, altering the thermal properties and requiring more energy to destroy the TPS-hybrid bonds. XRD analysis showed slight crystallinity changes, and TPS-15 hybrid showed an amorphous structure that allows the diffusion of EO thought the films. TPS-hybrid films showed antimicrobial behavior against E. coli due to the synergistic effect of MMT and EO, which resulted in strong antimicrobial effects due to the destabilization and partial destruction of the bacteria cell membrane. The hybrid probably changed the E. coli cell permeability, respiration, and electron transport. TPS-15 hybrid sample presented the lowest crystallinity, facilitating the film's oil diffusion, maximizing the bacterium inhibitory effect, and acting as a biocidal agent. The results demonstrated that a hybrid compound is a good option for starch antimicrobial films for fresh food, such as fruits and vegetables.

Fabrication and characterization of pullulan-based nanocomposites reinforced with montmorillonite and tempo cellulose nanofibril.

Nanocomposite film of pullulan (PULL), tempo cellulose nanofibrils (TOCNs) and, montmorillonite clay (MMT) were prepared using a solution casting method with aqueous solutions. X-ray diffraction data revealed that exfoliated MMT nanoplatelets are distributed within the PULL/TOCNs/MMT film structure. Fourier-transform infrared results revealed that there might be interactions among the TOCNs, MMT and PULL matrix led to improved tensile strength, thermal stability, water barrier properties, and decrease moisture susceptibility while maintained reasonable transparency and biodegradability of the ternary PULL nanocomposites. These excellent properties of the nanocomposites clearly indicate towards a new strategy for developing high-performance PULL-based nanocomposites by using two different types of fillers with various geometric shapes and aspect ratio. This kind of ternary nanocomposite film can be broadly used in food packaging and protection as a green and biodegradable film.

Taguchi design-assisted immobilization of Candida rugosa lipase onto a ternary alginate/nanocellulose/montmorillonite composite: Physicochemical characterization, thermal stability and reusability studies.

Biomass from oil palm frond leaves (OPFL) is an excellent reservoir of lignocellulosic material which full potential remains untapped. This study aimed to statistically optimize the covalent immobilization of Candida rugosa lipase (CRL) onto a ternary support comprised of OPFL derived nanocellulose (NC) and montmorillonite (MMT) in alginate (ALG) (CRL-ALG/NC/MMT). The coarser topology and the presence of characteristic spherical globules in the field emission scanning electron micrographs and atomic force micrographs, respectively, supported the existence of CRL on ALG/NC/MMT. In addition, amide peaks at 3478 and 1640 cm-1 in the fourier transform infrared spectra affirmed that CRL was covalently bonded to ALG/NC/MMT. The optimized Taguchi Design-assisted immobilization of CRL onto ALG/NC/MMT (7 h of immobilization, 35℃, pH 5, 7 mg/mL protein loading) gave a production yield of 92.89 % of ethyl levulinate (EL), as proven by gas chromatography-mass spectrometric ([M] +m/z 144, C7H12O3), FTIR and nuclear magnetic resonance (CAS-539-88-8) data. A higher optimal reaction temperature (50℃) and the reusability of CRL-ALG/NC/MMT for up to 9 esterification cycles substantiated the appreciable structural rigidification of the biocatalyst by ALG/NC/MMT, which improved the catalytic activity and thermal stability of the lipase.

Dietary montmorillonite as growth promoter and immunomodulator in rainbow trout (Oncorhynchus mykiss).

A 98-day research was conducted to assess the influence of dietary montmorillonite (MMT) on growth and immune competence in rainbow trout (Oncorhynchus mykiss). Fish fry (mean weight: 500 ± 35 mg) were fed with four experimental diets contained MMT at levels of 0 (as control group), 1, 2 and 4%. At the end of the experiment, the survival rate and growth parameters and serum total protein and albumin levels in MMT experimental groups was higher than the control group. Regarding humoral immune responses, fish being fed with 2 and 4% MMT diets showed the greatest and the lowest serum C3, C4 and lysozyme levels, respectively. Serum alternative complement pathway activity (ACH50) in groups fed with 1 and 2% MMT supplemented diets was higher than the other groups. At the end of the challenge test with VHSV, the mortality rate reached to 28 and 20% in the control group and 4% MMT supplemented groups, respectively, whereas fish fed with 1 and 2% MMT supplemented diets showed the lowest mortality rate (2.5%) Fish fed with 1 and 2% MMT supplemented groups showed higher head kidney IL-1ß and TNF-α genes expression compared to the other treatment groups. In addition, fish fed with 2% MMT-supplemented diet had the highest head kidney Lyz-2 gene expression. In summary, the results of this research suggests that supplementing diet with MMT at 1 or 2% improve growth and non-specific humoral immune responses and induce head kidney immune relate genes expression and disease resistance against VHS in O. mykiss.