ABSTRACT
Adlay bran is known for its nutrient-rich profile and multifunctional properties, and steam explosion (SE) is an emerging physical modification technique. However, the specific effects of SE on the activity composition and antioxidant capacity of adlay bran soluble dietary fiber (SDF) during in vitro digestion, as well as its influence on gut microbiota during in vitro fermentation, remain inadequately understood. This paper reports the in vitro digestion and fermentation characteristics of soluble dietary fiber from adlay bran modified by SE (SE-SDF). Compared with the untreated samples (0-SDF), most of the phenolic compounds and antioxidant capacity were significantly increased in the SE-SDF digests. Additionally, SE was beneficial for adlay bran SDF to increase the content of acetic acid, propionic acid and total short-chain fatty acids (SCFAs) in fermentation broth during in vitro fermentation. SE-SDF could promote the growth of beneficial bacteria while inhibiting the proliferation of pathogenic microbes. Our research indicates that SE-SDF shows strong antioxidant properties after in vitro digestion and plays a pivotal role in regulating gut microbiota during in vitro fermentation, ultimately enhancing human intestinal health.
Subject(s)
Antioxidants , Coix , Dietary Fiber , Digestion , Fatty Acids, Volatile , Fermentation , Gastrointestinal Microbiome , Steam , Dietary Fiber/metabolism , Gastrointestinal Microbiome/physiology , Antioxidants/metabolism , Antioxidants/analysis , Fatty Acids, Volatile/metabolism , Fatty Acids, Volatile/analysis , Coix/chemistry , Humans , Propionates/metabolism , Food Handling/methodsABSTRACT
As a member of the neonicotinoid group, imidaclothiz has garnered increasing attention due to its possible health risks. This study investigated the metabolism and distribution of imidaclothiz in mice. Seven imidaclothiz metabolites were found, four of which are known, and three are unknown. The metabolic reactions observed were hydroxylation, nitrate ester hydrolysis, methylation, urea formation, and reduction to NO. Precise quantification revealed that after 2 h of oral administration, imidaclothiz rapidly dispersed into various organs and tissues, peaking at 4 h, and was then swiftly eliminated. No propensity for accumulation in the body, particularly in the liver, was observed. Toxicity data from the T.E.S.T prediction indicated that imidaclothiz had moderate toxicity to rats, and a majority of its metabolites were more toxic than the parent compound. These findings complement the existing knowledge of the imidaclothiz environmental fate in mammals and offer a reference point for its application in agriculture and industry.
Subject(s)
Insecticides , Neonicotinoids , Animals , Mice , Insecticides/metabolism , Neonicotinoids/metabolism , Male , Liver/metabolism , Liver/drug effects , Rats , FemaleABSTRACT
Sterigmatocystin (STC) is an emerging mycotoxin that poses a significant threat to the food security of cereal crops. To mitigate STC contamination in maize, this study employed selected lactic acid bacteria as biocontrol agents against Aspergillus versicolor, evaluating their biocontrol potential and analyzing the underlying mechanisms. Lactiplantibacillus plantarum HJ10, isolated from pickle, exhibited substantial in vitro antifungal activity and passed safety assessments, including antibiotic resistance and hemolysis tests. In vivo experiments demonstrated that L. plantarum HJ10 significantly reduced the contents of A. versicolor and STC in maize (both >84 %). The impact of heat, enzymes, alkali, and other treatments on the antifungal activity of cell-free supernatant (CFS) was investigated. Integrated ultra-high-performance liquid chromatography (UPLC) and gas chromatography-mass spectrometry (GC-MS) analysis revealed that lactic acid, acetic acid, and formic acid are the key substances responsible for the in vitro antifungal activity of L. plantarum HJ10. These metabolites induced mold apoptosis by disrupting cell wall structure, increasing cell membrane fluidity, reducing enzyme activities, and disrupting energy metabolism. However, in vivo antagonism by L. plantarum HJ10 primarily occurs through organic acid production and competition for growth space and nutrients. This study highlights the potential of L. plantarum HJ10 in reducing A. versicolor and STC contamination in maize.
Subject(s)
Aspergillus , Lactobacillales , Sterigmatocystin , Zea mays , Zea mays/microbiology , Aspergillus/metabolism , Aspergillus/growth & development , Lactobacillales/metabolism , Antifungal Agents/pharmacology , Food Contamination/prevention & control , AntibiosisABSTRACT
Mycotoxins are secondary fungal metabolites harmful to humans and animals. Patulin (PAT) is a toxin found in different food products but especially in apples and their derivative products. The most common fungi producers of this compound are Aspergillus clavatus and Penicillium expansum. The production of patulin, as other mycotoxins, can be impacted by diverse phenomena such as water and nutrient availability, UV exposure, and the presence of antagonistic organisms. Consequently, gaining a comprehensive understanding of climate and environmental conditions is a crucial step in combating patulin contamination. In this study, moulds were isolated from 40 apple samples collected from seven locations across Hungary: Csenger, Damak, Pallag, Lövopetri, Nagykálló, and Újfehértó. A total of 183 moulds were morphologically identified, with 67 isolates belonging to the Alternaria, 45 to the Aspergillus, and 13 to the Penicillium groups. The location possessed a higher influence than farming method on the distribution of mould genera. Despite the requirement of higher temperature, Aspergillus species dominated only for the region of Újfehértó with approximately 50% of the isolates belonging to the genus. Four of the seven locations assessed: Csenger, Debrecen-Pallag, Nyírtass and Nagykálló, were dominated by Alternaria species. All isolates belonging to the genera Aspergillus and Penicillium were tested for the presence of the isoepoxidone dehydrogenase (idh) gene, a key player in the patulin metabolic pathway. To guarantee patulin production, this ability was confirmed with TLC assays. The only Aspergillus strain that presented a positive result was the strain Aspergillus clavatus B9/6, originated from the apple cultivar Golden Reinders grown in Debrecen-Pallag by integrated farming. Of the Penicillium isolates only one strain, B10/6, presented a band of the right size (500-600 bp) for the idh gene. Further sequencing of the ITS gene showed that this strain should be classified as Talaromyces pinophilus. The TLC tests confirmed this microorganism as the only patulin producer under the studied conditions for its cluster.
Subject(s)
Aspergillus , Malus , Patulin , Penicillium , Patulin/analysis , Penicillium/metabolism , Penicillium/isolation & purification , Malus/chemistry , Malus/microbiology , Aspergillus/metabolism , Aspergillus/isolation & purification , Aspergillus/chemistry , Hungary , Food Contamination/analysis , Food MicrobiologyABSTRACT
In this study, a QuEChERS method based on citrate was developed and utilized for the analysis of twelve neonicotinoid pesticides in fresh red chilies, fresh green chilies, and dried chilies, coupled with ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS). In the sample preparation, acetonitrile containing 1% formic acid was used as the extraction solvent. Anhydrous sodium sulfate replaced the traditional anhydrous magnesium sulfate for water removal, effectively eliminating the issues of salt caking. Graphitized carbon black, octadecyl silica, and primary secondary amine were used as cleaning agents. The method showed good sensitivity, with the limits of quantification below 0.03 mg/kg for fresh chilies and below 0.15 mg/kg for dried chilies. Values of matrix effects ranged from -19.5% to 8.4%, and the recovery was 86.9% - 105.2%. The analytical method provided an effective tool for the high throughput detection of neonicotinoid pesticide residues in multiple chili matrices.
Subject(s)
Capsicum , Food Contamination , Pesticide Residues , Chromatography, High Pressure Liquid , Capsicum/chemistry , Food Contamination/analysis , Pesticide Residues/analysis , Pesticide Residues/chemistry , Pesticide Residues/isolation & purification , Mass Spectrometry/methods , Neonicotinoids/analysis , Neonicotinoids/chemistry , Tandem Mass Spectrometry/methodsABSTRACT
Identifying aflatoxin-detoxifying probiotics remains a significant challenge in mitigating the risks associated with aflatoxin contamination in crops. Biological detoxification is a popular technique that reduces mycotoxin hazards and garners consumer acceptance. Through multiple rounds of screening and validation tests, Geotrichum candidum XG1 demonstrated the ability to degrade aflatoxin B1 (AFB1) by 99-100%, exceeding the capabilities of mere adsorption mechanisms. Notably, the degradation efficiency was demonstrably influenced by the presence of copper and iron ions in the liquid medium, suggesting a potential role for proteases in the degradation process. Subsequent validation experiments with red pepper revealed an 83% reduction in AFB1 levels following fermentation with G. candidum XG1. Furthermore, mass spectrometry analysis confirmed the disruption of the AFB1 furan ring structure, leading to a subsequent reduction in its toxicity. Collectively, these findings establish G. candidum XG1 as a promising candidate for effective aflatoxin degradation, with potential applications within the food industry.
Subject(s)
Aflatoxin B1 , Food Contamination , Geotrichum , Probiotics , Aflatoxin B1/metabolism , Aflatoxin B1/chemistry , Aflatoxin B1/analysis , Probiotics/metabolism , Probiotics/chemistry , Geotrichum/metabolism , Geotrichum/chemistry , Food Contamination/analysis , Fermentation , Capsicum/chemistry , Capsicum/metabolism , Capsicum/microbiology , ChinaABSTRACT
Chlorophylls and ß-carotene are fat-soluble phytochemicals in daily diets, while their bioaccessibility interaction remains unknown. Eight dietary chlorophylls and their derivatives (chlorophyll a, chlorophyll b, pheophytin a, pheophytin b, chlorophyllide a, chlorophyllide b, pheophorbide a, pheophorbide b) were combined with ß-carotene in six different oil matrices (corn oil, coconut oil, medium-chain triglycerides, peanut oil, olive oil and fish oil) and were subjected to in vitro digestion. Generally, chlorophylls significantly decreased ß-carotene bioaccessibility by competitive incorporation into micelles. Dephytylated chlorophylls had a greater inhibitory effect on the micellarization and bioaccessibility of ß-carotene compared to phytylated chlorophylls. In their co-digestion system, olive oil group exhibited the smallest particle size and biggest zeta potential in both digesta and micelles. For chlorophylls, the phytol group and their levels are key factors, which was also buttressed by the mice model where additional supplementation of pheophorbide a significantly hindered the accumulation of ß-carotene and retinoids compounds.
Subject(s)
Chlorophyll , beta Carotene , Chlorophyll/chemistry , Chlorophyll/metabolism , beta Carotene/chemistry , beta Carotene/metabolism , Animals , Mice , Biological Availability , Digestion , Humans , Plant Oils/chemistry , Plant Oils/metabolism , Models, Biological , MicellesABSTRACT
Aspergillus flavusï¼A. flavusï¼, a common humic fungus known for its ability to infect agricultural products, served as the subject of investigation in this study. The primary objective was to assess the antifungal efficacy and underlying mechanisms of binary combinations of five volatile organic compounds (VOCs) produced by lactic acid bacteria, specifically in their inhibition of A. flavus. This assessment was conducted through a comprehensive analysis, involving biochemical characterization and transcriptomic scrutiny. The results showed that VOCs induce notable morphological abnormalities in A. flavus conidia and hyphae. Furthermore, they disrupt the integrity of the fungal cell membrane and cell wall, resulting in the leakage of intracellular contents and an increase in extracellular electrical conductivity. In terms of cellular components, VOC exposure led to an elevation in malondialdehyde content while concurrently inhibiting the levels of total lipids, ergosterol, soluble proteins, and reducing sugars. Additionally, the impact of VOCs on A. flavus energy metabolism was evident, with significant inhibition observed in the activities of key enzymes, such as Na+/K+-ATPase, malate dehydrogenase, succinate dehydrogenase, and chitinase. And they were able to inhibit aflatoxin B1 synthesis. The transcriptomic analysis offered further insights, highlighting that differentially expressed genes (DEGs) were predominantly associated with membrane functionality and enriched in pathways about carbohydrate and amino acid metabolism. Notably, DEGs linked to cellular components and energy-related mechanisms exhibited down-regulation, thereby corroborating the findings from the biochemical analyses. In summary, these results elucidate the principal antifungal mechanisms of VOCs, which encompass the disruption of cell membrane integrity and interference with carbohydrate and amino acid metabolism in A. flavus.
Subject(s)
Antifungal Agents , Aspergillus flavus , Volatile Organic Compounds , Volatile Organic Compounds/pharmacology , Aspergillus flavus/drug effects , Aspergillus flavus/metabolism , Antifungal Agents/pharmacology , Lactobacillales/metabolismABSTRACT
Serratia marcescens strains from a dairy-producing environment were tested for their inhibitory effect on Listeria monocytogenes, Salmonella Hartford, Yersinia enterocolitica and Escherichia coli. Inhibition of foodborne pathogens was observed in the case of a non-pigmented Serratia strain, while the pigment-producing isolate was able to inhibit only Y. enterocolitica. The co-culturing study in tryptone soya broth (TSB) and milk showed that the growth of Salmonella was inhibited in the first 24 h, but later the pathogen could grow in the presence of the Serratia strain even if its cell concentration was 1000 times higher than that of Salmonella. However, we found that (1) concentrated cell-free supernatants had stronger inhibitory activity, which confirms the extracellular nature of the antagonistic compound(s). We proved that (2) protease and chitinase enzymes can take part in this mechanism, but they are not the main inhibitory compounds. The presence of prodigiosin was observed only in the case of the pigmented strain; thus, (3) we hypothesized that prodigiosin does not take part in the inhibition of the pathogens. However, (4) the combined effect of different extracellular metabolites might be attributed to the inhibitory property. Application of concentrated S. marcescens cell-free supernatant can be an effective antibacterial strategy in the food industry, mainly in the form of a bio-disinfectant on surfaces of food-processing areas.
ABSTRACT
To explore the antifungal mechanisms of volatile organic compounds (VOCs) produced by Pseudomonas fluorescens ZX against Botrytis cinerea, biochemical analyses and transcriptomic techniques were employed in this work. The results showed that P. fluorescens ZX-producing VOCs can increase the cell membrane permeability of B. cinerea and disrupt cell membrane integrity, resulting in leakage of the pathogen's cellular contents, inhibition of ergosterol biosynthesis (about 76%), and an increase in malondialdehyde (MDA) content. Additionally, for B. cinerea respiration, P. fluorescens ZX-producing VOCs (1 × 109 CFU /mL) significantly inhibited the activities of ATPase (55.7%), malate dehydrogenase (MDH) (33.1%), and succinate dehydrogenase (SDH) (57.9%), seriously interfering with energy metabolism and causing accumulation of reactive oxygen species (ROS). Furthermore, transcriptome analysis of B. cinerea following exposure to VOCs revealed 4590 differentially expressed genes (DEGs) (1388 upregulated, 3202 downregulated). Through GO analysis, these DEGs were determined to be enriched in intrinsic components of membrane, integral components of membrane, and membrane parts, while KEGG analysis indicated that they were enriched in many amino acid metabolism pathways. Significantly, the DEGs related to ergosterol biosynthesis, ATPase, mitochondrial respiratory chain, malate dehydrogenase, and cell membrane showed down-regulation, corroborating the biochemical analyses. Taken together, these results suggest that the antifungal activity of P. fluorescens ZX-producing VOCs against B. cinerea occurs primary mechanisms: causing significant damage to the cell membrane, negatively affecting respiration, and interfering with amino acid metabolism.
Subject(s)
Antifungal Agents , Pseudomonas fluorescens , Volatile Organic Compounds , Adenosine Triphosphatases/metabolism , Amino Acids/metabolism , Antifungal Agents/chemistry , Antifungal Agents/metabolism , Botrytis , Ergosterol/metabolism , Malate Dehydrogenase/metabolism , Plant Diseases/microbiology , Pseudomonas fluorescens/chemistry , Pseudomonas fluorescens/metabolism , Volatile Organic Compounds/pharmacology , Volatile Organic Compounds/metabolismABSTRACT
The preparation of dephytylated chlorophyll standards is inefficient and the process is complicated, which hinders chlorophyll determination and related bioactive property investigation. In this paper, chlorophyll derivatives from four phytylated chlorophylls (chlorophyll a, chlorophyll b, pheophytin a, and pheophytin b) before and after the enzymatic reaction were qualitatively and quantitatively characterized by UPLC-DAD-MS. A simple index was proposed to characterize chlorophyll pigments from their oxidized counterparts by the λmax of the typical peak of chlorophyll derivatives in UV-visible spectrum and their signal intensity ratios. The optimal reaction conditions for the enzymatic reaction of four chlorophyll pigments were optimized, and kinetic models were fitted. The results showed that the optimal temperatures for the enzymatic reactions of chlorophyll a, chlorophyll b, pheophytin a, and pheophytin b were 30, 30, 60, and 60°C, respectively, and their optimal reaction time was 2, 3, 1, and 3 h, respectively. Kinetic models were fitted under optimal reaction conditions to study the Km and Vm values of the enzymatic reactions. PRACTICAL APPLICATION: Dephytylated chlorophylls, such as chlorophyllide and pheophorbide, are frequently determined in food industry and are always required to be prepared in lab with acetone powder from plant tissue. Moreover, chlorophyll pigments are easy to undergo oxidations, which make the characterization of dephytylated chlorophyll pigments more complicated and difficult. In this paper, four types of phytylated chlorophylls were investigated respectively about the dephytinization process with the citrus acetone powder, and the reaction mixture was analyzed with UPLC-DAD-MS, which can provide an important reference for relevant chlorophyll determination studies and the development of chlorophyll identification protocols.
Subject(s)
Acetone , Pheophytins , Chlorophyll A , Powders , ChlorophyllABSTRACT
Brocade orange (Citrus sinensis L. Osbeck) peel, a by-product which is usually discarded in large amounts, is rich in a broad spectrum of phenols. Accordingly, this study investigated the distribution and natural variation of free, esterified, glycosylated and insoluble-bound phenols (FPs, EBPs, GBPs, and IBPs) in the peels. Regardless of phenolic fraction or peel position, the total phenol and flavonoid contents, and most tested phenolic compound contents were generally abundant during the immature and semi-mature stages but existed at lower levels during the commercial mature period. The flavedo was much richer than the albedo in a few phenolic acids, flavonols, flavones, and especially polymethoxyflavonoids, which was particularly true for EBPs. Flavanones, particularly in GBP form, were generally present in equal or even much higher levels in the albedo. The four phenolic forms exhibited distinct trends in terms of abundance. In the flavedo (except the young fruit stage) followed the order: EBPs > GBPs ≈ FPs â« IBPs, and in the albedo: GBPs â« FPs ≈ EBPs â« IBPs. Generally, the phenols examined for this study were highly abundant in the citrus peels, endowing this agricultural waste with great potential to be an excellent natural source of functional ingredients.
Subject(s)
Citrus sinensis , Citrus , Flavonoids , Fruit/chemistry , Phenols/analysisABSTRACT
A digestibility trial was conducted with African catfish hybrid juveniles in order to determine the apparent digestibility coefficients (ADCs) of different nutrients. The experimental diets contained defatted black soldier fly (BSL), yellow mealworm (MW), or fully fat blue bottle fly (BBF) meals, in a 70 : 30 ratio between the control diet and the tested insect meals. The indirect method for the digestibility study was performed using 0.1% yttrium oxide as an inert marker. Fish juveniles of 217.4 ± 9.5 g initial weight were distributed in 1 m3 tanks (75 fish/tank) of a recirculating aquaculture system (RAS), in triplicates, and fed until satiation for 18 days. The average final weight of the fish was 346 ± 35.8 g. The ADCs of the dry matter, protein, lipid, chitin, ash, phosphorus, amino acids, fatty acids, and gross energy for the test ingredients and diets were calculated. A six-month storage test was carried out to evaluate the shelf life of the experimental diets, while the peroxidation and microbiological status of the diets were also assessed. The ADC values of the test diets differed significantly (p < 0.001) compared to those of the control for most of the nutrients. Altogether, the BSL diet was significantly more digestible for protein, fat, ash, and phosphorus than the control diet but less digestible for essential amino acids. Significant differences were found between the ADCs of the different insect meals evaluated (p < 0.001) for practically all nutritional fractions analyzed. The African catfish hybrids were able to digest BSL and BBF more efficiently than MW, and the calculated ADC values agreed with those of other fish species. The lower ADCs of the tested MW meal correlated (p < 0.05) with the markedly higher acid detergent fiber (ADF) levels present in the MW meal and MW diet. Microbiological evaluation of the feeds revealed that mesophilic aerobic bacteria in the BSL feed were 2-3 orders of magnitude more abundant than those in the other diets and their numbers significantly increased during storage. Overall, BSL and BBF proved to be potential feed ingredients for African catfish juveniles and the shelf life of the produced diets with 30% inclusion of insect meal retained the required quality during a six-month period of storage.
ABSTRACT
Nowadays, demand for products which beyond the overall nutritional value have a feature that protects the consumers health, have increased. Several studies have proved fruit juices can become suitable carrier or medium for probiotic organisms. Therefore the aim of our study was to investigate the possibility of the probiotication of sour cherry juice by lactic acid fermentation with probiotic starter culture. In the fermentation 9 Lactobacillus strains were used and two cultivars of sour cherry as raw material. The pH adjustment and supplement of nutrients were necessary and to reach the recommended probiotic cell count we also investigated the effect of dilution of sour cherry juice. Due to the optimized combination of the pH adjustment, supplementation and dilution, the investigated strains reached the desired 9 log cfu/mL cell density in sour cherry juices, however a significant difference was observed between the number of viable cells of some Lactobacillus strains. In the Újfehértói fürtös sour cherry L. acidophilus La-5 (9.43 log cfu/mL), while in the Petri species L. acidophilus 150 (9.60 log cfu/mL) resulted in the highest probiotic cell number. The lactic acid fermentation can increase the phenolic compounds, but in case of the bioactive compounds significant differences were not general between the strains.
Subject(s)
Probiotics , Prunus avium , Fermentation , Fruit and Vegetable Juices , Lactic Acid , Lactobacillus , Prunus avium/chemistryABSTRACT
Preharvest application of biocontrol agents is a promising strategy for promoting biosynthesis and accumulation of functional ingredients in fruit crops. In this study, we sought to evaluate the potential of Pseudomonas fluorescens ZX in stimulating the primary and secondary metabolism of citrus fruit peel. Pretreatment with P. fluorescens ZX was found to significantly affect the concentrations and profiles of both primary and secondary metabolites. More importantly, using P. fluorescens ZX suspension to increase inoculation numbers during fruit development typically elicited stronger stimulus effects, and multiple applications of P. fluorescens ZX significantly improved the biosynthesis process of beneficial compounds, resulting in their abundant accumulation in the peel. In fruit pretreated four times with P. fluorescens ZX, hesperidin, sinensetin, nobiletin, synephrine, and pectin were increased by approximately 26.0, 31.3, 44.8, 19.7, and 23.1%, respectively, compared to the untreated control. Collectively, these results indicated that, as a biostimulant, preharvest application of P. fluorescens ZX is an effective, affordable, ecological, and ecofriendly alternative agricultural technique for exploiting citrus crops. This approach is also promising for increasing the value of citrus fruit peel (currently regarded primarily as processing waste), thereby allowing industrial agricultural practices to move one step closer toward a circular economy.
Subject(s)
Citrus , Hesperidin , Pseudomonas fluorescens , Fruit , Pseudomonas fluorescens/genetics , Secondary MetabolismABSTRACT
In this study, volatile organic compounds (VOCs) were generated by Pseudomonas fluorescens ZX with incubation in nutrient broth (NB), on NA (NB with agar), and on healthy orange fruits, and pure individual components of VOCs were used to manage citrus green mold infected by Penicillium digitatum. At a concentration of 1 × 1010 cfu/mL, the VOCs from antagonist-containing NA plates inhibited P. digitatum conidial germination and mycelial growth by about 60%, while the VOCs from bacterial fluid exhibited approximately 75% inhibitory effect. Biofumigation by VOCs significantly reduced the disease index, with a higher biocontrol efficacy by VOCs from bacterial fluid (about 51%) than from antagonist-containing NA plates (around 40%) or from antagonist-infested fruit (approximately 24%). Exposure to VOCs led to morphological abnormalities of P. digitatum conidia and hyphae. However, VOCs exhibited poor preventative and curative action against P. digitatum. The storage test showed that biofumigation had no negative effects on fruit quality. Antifungal assays suggested that dimethyl disulfide and dimethyl trisulfide exhibited the highest inhibitory effects, which afforded complete inhibition at the lowest concentrations. In addition, organic acids were also promising in controlling green mold, but only at suitable low concentrations to avoid eliciting fruit's physiological diseases.
Subject(s)
Citrus , Penicillium , Pseudomonas fluorescens , Volatile Organic Compounds , Fruit , Plant DiseasesABSTRACT
Lactobacillus plantarum KFY02 (KFY02), isolated from naturally fermented milk yoghurt in Korla, Xinjiang, Northwest of China, showed gardenoside action for the intestinal regulation of constipated mice. Comparatively, the effects of KFY02 (0.5 × 108 CFU kg-1, by body weight (BW)), gardenoside (50 mg kg-1, BW), and KFY02 (0.5 × 108 CFU kg-1, BW) + gardenoside (50 mg kg-1, BW) on intestinal regulation in mice with montmorillonite-induced constipation were also studied. Enzyme linked immunoassay, hemotoxylin and eosin (H&E) staining, quantitative polymerase chain reaction (qPCR) assay and high performance liquid chromatography (HPLC) analysis were used for the study. Compared with the model group, KFY02 + genipin (combined group) increased the propelling rate of activated carbon in the small intestines of mice and accelerated the discharge of the first black stool in mice. At the same time, the combination group reduced the levels of motilin (MTL), substance P (SP) and endothelin-1 (ET-1) in the serum, and increased the somatostatin (SS), vasoactive intestinal peptide (VIP), acetylcholinesterase (AchE) and gastrin (Gastrin) levels in the serum, which made these parameters close to those of the normal group. Using qPCR analysis, it was observed that the combined group up-regulated the mRNA expression of endothelial nitric oxide synthase (eNOS), stem cell factor (SCF), stem cell factor receptor (c-Kit), glutathione (GSH), catalase and manganese-superoxide dismutase (Mn-SOD) and down-regulated the expression of inducible nitric oxide synthase (iNOS) and transient receptor potential vanilloid receptor 1 (TRPV1). The combination increased the Bacteroides and Akkermansia abundances and decreased the Firmicutes abundance in the feces of the constipated mice and decreased the Firmicutes/Bacteroides ratio. The expression of the above genes was similar to that of the normal group. The results indicate that KFY02 produced ß-glucosidase to hydrolyze the gardenoside glycosidic bond to produce genipin and can effectively promote the regulation of gastrointestinal hormones and intestinal peristalsis and reduce oxidative tissue damage in constipated mice. This study also confirmed that KFY02 has similar relieving effects to gardenoside for constipation in mice.
ABSTRACT
BACKGROUND: The application of chemical fungicides is currently the main method for the control of postharvest decay of fruits and vegetables. However, public concern has been growing towards the effect of fungicides on food safety, human health and environmental pollution. Thus, interest in microbial biocontrol agent development has grown, such agents being considered both safe and environmentally friendly. Pseudomonas fluorescens is widely distributed in nature, and one of the most valuable biocontrol and plant growth-promoting rhizobacteria. In this study, the efficacy and the potential associated modes of action of P. fluorescens ZX against Penicillium italicum on oranges (Citrus sinensis Osbeck) were investigated. RESULTS: The application of P. fluorescens ZX significantly reduced blue mold lesion size and incidence in comparison to the control, where P. fluorescens ZX was effective when applied preventatively but not curatively. In dual cultures, treatment with cell-free autoclaved cultures or culture filtrate had a limited capacity to suppress P. italicum, while P. italicum was inhibited by bacterial fluid and bacterial suspension with living cells in vitro. The P. fluorescens ZX isolate displayed protease, but not chitinase, glucanase or cellulose, activity, and produced siderophores and volatile organic compounds with antifungal abilities. Competition tests showed P. fluorescens ZX could use fructose, sucrose, aspartic acid, threonine, serine, glycine, valine, lysine and proline better than P. italicum. Furthermore, an effective biofilm that peaked after a 24-hour incubation at 30 °C was formed by the P. fluorescens ZX isolate. Light microscopy and scanning electron microscopy observations indicate the P. fluorescens ZX isolate could not undergo direct parasitism or hyperparasitism. CONCLUSIONS: Competition for nutrients and niches, biofilm formation, inhibition of spore germination and mycelial growth, and production of inhibitory metabolites may play important roles in P. fluorescens ZX antagonism of P. italicum. © 2019 Society of Chemical Industry.
Subject(s)
Antibiosis , Citrus/microbiology , Pseudomonas fluorescens/physiology , Biofilms , Fruit/microbiology , Mycelium/growth & development , Mycelium/metabolism , Penicillium/growth & development , Penicillium/metabolism , Plant Diseases/microbiology , Plant Diseases/prevention & controlABSTRACT
Colitis severely affects the quality of life of patients, and lactic acid bacteria have been reported to be able to improve or treat colitis. In this study, we selected a strain of Lactobacillus fermentum (CQPC04) with good resistance in vitro to evaluate its effect on improvement in mice with dextran sulfate sodium (DSS)-induced colitis. We analyzed the effects of L. fermentum CQPC04 on mice with colitis macroscopically via colon length and histopathology. We also used conventional biochemical and ELISA kits, real-time quantitative PCR (RT-qPCR), and Western blotting to analyze microscopically the effects of L. fermentum CQPC04 on related oxidant indices and pro- and anti-inflammatory cytokines in serum and colon tissue of mice. The results indicated that L. fermentum CQPC04 notably increased colon length and ameliorated pathological damage of colon tissue in colitic mice. Serum indices showed that L. fermentum CQPC04 increased the enzyme activity of total superoxide dismutase (T-SOD) and catalase (CAT) and decreased the content of malondialdehyde (MDA) and the activity of myeloperoxidase (MPO). In addition, it inhibited the release of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), IFN-γ, IL-1ß, IL-6, and IL-12, and increased the release of the anti-inflammatory cytokine IL-10 in serum. The RT-qPCR experiments confirmed that L. fermentum CQPC04 downregulated the expression of pro-inflammatory cytokine nuclear factor-κB-p65 (NF-κBp65), NF-κB inhibitor-α (IκB-α), TNF-α, IFN-γ, IL-1ß, IL-6, cyclooxygenase 2 (COX-2), and inducible nitric oxide synthase (iNOS), and upregulated the expression of IL-10 in colon tissue. Western blot analysis indicated that L. fermentum CQPC04 significantly reduced expression of NF-κBp65, TNF-α, IL-1ß, COX-2, and iNOS in mouse colon tissues, and increased expression of IκB-α and superoxide dismutase 2 (SOD2). Thus, L. fermentum CQPC04 could effectively alleviate the symptoms of DSS-induced colitis mice and is a potential probiotic for human experiments.
Subject(s)
Colitis/diet therapy , Limosilactobacillus fermentum , NF-kappa B/metabolism , Probiotics/therapeutic use , Signal Transduction/drug effects , Animals , Colitis/chemically induced , Colitis/metabolism , Colitis/prevention & control , Cytokines/blood , Dextran Sulfate , Female , Mice , Mice, Inbred ICR , Nitric Oxide Synthase Type II/metabolism , Protective Agents , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Optical waveguide lightmode spectroscopy (OWLS) technique has been applied to label-free detection of aflatoxin B1 in a competitive immunoassay format, with the aim to compare the analytical goodness of the developed OWLS immunosenor with HPLC and enzyme-linked immunosorbent assay (ELISA) methods for the detection of aflatoxin in spice paprika matrix. We have also assessed applicability of the QuEChERS method prior to ELISA measurements, and the results were compared to those obtained by traditional solvent extraction followed by immunoaffinity clean-up. The AFB1 content of sixty commercial spice paprika samples from different countries were measured with the developed and optimized OWLS immunosensor. Comparing the results from the indirect immunosensor to that obtained by HPLC or ELISA provided excellent correlation (with regression coefficients above 0.94) indicating that the competitive OWLS immunosensor has a potential for quick determination of aflatoxin B1 in paprika samples.