Synthetic Gastric Fluid Research Articles

Conversion of water-soluble CBD to Δ9-THC in synthetic gastric fluid - an unlikely cause of positive drug tests.

Cannabidiol (CBD) has been shown to convert to Δ9-tetrahydrocannabinol (Δ9-THC) in acidic environments, raising a concern of conversion when exposed to gastric fluid after consumption. Using synthetic gastric fluid (SGF), it has been demonstrated that the conversion requires surfactants, such as sodium dodecyl sulfate (SDS), due to limited solubility of CBD. Recently, water-compatible nano-emulsions of CBD have been prepared as a means of fortifying beverages and water-based foods with CBD. Since these emulsions contain surfactants as part of their formulation, it is possible that these preparations might enhance the production of Δ9-THC even in the absence of added surfactants. Three THC-free CBD products; an oil, an anhydrous powder, and a water-soluble formulation were incubated for 3 hours in SGF without SDS. The water-soluble CBD product produced a dispersion while the powder and the oil did not mix with the SGF. No THC was detected with the CBD oil (<0.0006% conversion), and up to 0.063% and 0.0045% conversion to Δ9-THC was observed with the water-soluble CBD and the CBD powder, respectively. No formation of Δ8-THC was observed. In comparison, when the nano-formulated CBD was incubated in SGF with 1% SDS, 33-36% conversion to Δ9-THC was observed. Even though the rate of conversion with the water-soluble CBD was at least 100-fold higher compared to the CBD oil, it was still smaller than Δ9-THC levels reported in CBD products labelled "THC-free" or "<0.3% THC" based on the Agricultural Improvement Act of 2018 (the Farm Bill). Assuming a daily CBD dose of around 30 mg/day, it is unlikely that conversion of CBD to Δ9-THC could produce a positive urinary drug test for Δ9-THC-COOH (15 ng/mL cutoff).

Impact of Elaeagnus angustifolia flour added to bio-yogurt on probiotic survival and monitoring of in vitro acid tolerance in synthetic gastric fluid

Impact of Elaeagnus angustifolia flour added to bio-yogurt on probiotic survival and monitoring of in vitro acid tolerance in synthetic gastric fluid

Ion-imprinted-based nanochelators for iron(III) removal from synthetic gastric fluid

Ion-imprinted-based nanochelators for iron(III) removal from synthetic gastric fluid

Determination of the changes in the gastric fluid endurance of O157 and non-O157 Shiga toxin-producing Escherichia coli during storage of experimentally produced beef frankfurter.

Resistance of Shiga toxin-producing Escherichia coli (STEC) O157:H7 and serogroups O103, O26 and O145 to synthetic gastric fluid (SGF, pH 1.5) were investigated during frankfurter storage. Pathogens were inoculated (5 ± 1 log10cfug-1) on frankfurters and frankfurters were stored at 4°C for 75days in vacuum packages. Population changes of the competitive flora and STEC, changes in the pH of the frankfurters and resistance of STEC to SGF were monitored on days 0, 15, 30, 45, 60 and 75 of frankfurter storage. Direct synthetic gastric fluid (DSGF) challenges were also conducted to assess pathogen resistance without being effected by frankfurters, by inoculating pathogen cultures directly into SGF. Results showed that acid resistance of O145 and O26 was stronger than that of O103 and O157 during frankfurter storage. Resistance of O103 to SGF was better than that of O157 during frankfurter storage but, was similar to that of O157 during DSGF challenges. Results indicate that acid resistance of some strains of STEC pathogens might differentiate during storage of frankfurters. Different resistance capabilities to SGF were observed in the STEC strains when inoculated and stored on frankfurters than directly inoculated in the SGF.

Survival of a norovirus surrogate on surfaces in synthetic gastric fluid

Norovirus (NoV) can survive in buffer at a range of pH values. However, when variations to pH are combined with the enzymatic conditions typical in gastric fluid, NoV survival is less predictable. Added to this, the material on which expelled vomitus fluid lands e.g. safety vinyl or carpet, might also affect the level of NoV survival in the environment. This study assessed survival of the NoV surrogate Feline calicivirus (FCV) after application onto four material types in the presence of synthetic gastric fluid of varying pH values. Synthetic gastric fluid of varying pH values (1.5, 2.5, 3.5 and 4.5) containing FCV (1×106 PFU/100 μL of gastric fluid) was seeded (100 μL) onto four material types: Formica, safety vinyl, cotton sheet and carpet tiles (2 cm × 2 cm) and left for 0, 30 and 60 minutes before recovering and identifying any viable viruses via plaque assay. FCV survived in synthetic gastric fluid at pH values as low as 2.5 and in some cases as low as 1.5 for at least 30 min when associated with certain materials. Greater average numbers of viable virus were recovered from the more absorbent materials of cotton (1900 PFU/sample) and carpet (1600 PFU/sample) compared with Formica (360 PFU/sample) and safety vinyl (380 PFU/sample). This study showed that FCV can survive in synthetic gastric fluids of low pH values for significant time periods especially on absorbent materials. This emphasises the importance of ensuring effective cleaning and decontamination procedures, particularly of soft furnishings and laundry, for infection prevention and control of NoV outbreaks.

Global transcriptomic response of Listeria monocytogenes during growth on cantaloupe slices

Understanding a pathogen's response to food environments is imperative to develop effective control strategies as well as to elucidate the impact of foods on virulence potential. The purpose of this study was to assess transcriptional response of Listeria monocytogenes after growth in cantaloupe, as well as its impact on survival in synthetic gastric fluid (SGF). The transcriptional profiles of L. monocytogenes grown in cantaloupe or Brain Heart Infusion (BHI) under refrigeration were compared by a custom-designed microarray. A total of 286 and 175 genes were significantly up- and down-regulated, respectively, in L. monocytogenes grown in cantaloupe as compared to BHI (fold change ≥ 2.5 and adj. P < 0.05). The majority of upregulated genes belonged to functions related to amino acid and nucleotide metabolism, flagellar biosynthesis, and iron acquisition, while most downregulated genes belonged to carbohydrate metabolism. Notably, the branched chain amino acid (BCAA: leucine, isoleucine, valine) biosynthesis operon was shown to be highly upregulated as well as the purine and pyrimidine biosynthesis pathways. Transcript levels of several stress- and virulence-related genes were significantly altered, implying an impact of growth in cantaloupe on the virulence potential of L. monocytogenes. Enhanced survival of L. monocytogenes in SGF following growth in cantaloupe further demonstrated the impact of cantaloupe-associated growth on the pathogen's subsequent response to a host relevant stress.

Separation of aflatoxin B1 from synthetic physiological fluids using talc and diatomite: Kinetic and isotherm aspects.

The objective of the study was to examine adsorption of the aflatoxin B1 from synthetic gastric fluid and synthetic intestinal fluid by talc, raw and calcined diatomite. The kinetic and equilibrium adsorption processes were studied in the batch adsorption experiments applying high performance liquid chromatography for the aflatoxin B1 determination. The kinetic study showed a very fast adsorption of the aflatoxin B1 onto the selected adsorbents from the both physiological fluids with reaching equilibrium within 1-15min. The aflatoxin B1 was almost completely adsorbed in initial linear step of the kinetic process that can be described well by the zero-order kinetics model. The experimental data of the equilibrium adsorption were characterized using the Langmuir and Freundlich isotherm models. The high adsorption effectiveness was found in a range of 90%-100% and 60%-100% for the diatomite samples and the talc respectively at the initial concentrations of the aflatoxin B1 as 31-300ng/mL. The possible mechanisms of the aflatoxin adsorption onto the used mineral adsorbents are also discussed in the work.

On-line dynamic extraction system hyphenated to inductively coupled plasma optical emission spectrometry for automatic determination of oral bioaccessible trace metal fractions in airborne particulate matter.

For a realistic evaluation of the potential hazard emanating from airborne particulate matter (APM), the determination of the total inhaled metal amounts associated with APM is insufficient in risk assessment. Additional information about metal fractions that can be mobilized by the human body is necessary, because only those soluble (also called bioaccessible) fractions can be absorbed by the human body, and thus potentially cause adverse health effects. In the present study, a dynamic flow-through approach as a front end to inductively coupled plasma optical emission spectrometry (ICP-OES) exploiting advanced flow analysis is employed for on-line handling of multiple APM samples and determination of bioaccessible trace metals under worst case extraction scenarios. The method is based on on-line continuous extraction of filter samples with synthetic gastric fluid followed by on-line ICP-OES measurement of the dissolved fraction of trace metals. The assembly permits an automated successive measurement of three sample replicates in less than 19min. The on-line extraction procedure offers increased sample throughput and reduced risk of sample contamination and overcomes metal re-adsorption processes compared to the traditional batch-wise counterparts. Furthermore, it provides deeper information on the kinetics of the leaching process. The developed procedure was applied to the determination of bioaccessible metal fractions (Al, Ba, Cu, Fe and Mn as model analytes) in PM10 samples from Palma de Mallorca (Spain) and Vienna (Austria). Graphical Abstract On-line gastric bioaccessibility of elements in airborne particulate matter.

Survival and expression of acid resistance genes in Shiga toxin-producing Escherichia coli acid adapted in pineapple juice and exposed to synthetic gastric fluid.

The aim of this research was to determine the ability of non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups to survive with exposure to synthetic gastric fluid (SGF) after adaptation to pineapple juice (PJ) at room and refrigerated temperatures compared to E.coli O157:H7 and to examine the relative transcriptional expression of acid resistance (AR) genes, rpoS, gadA and adiA. Resistant and sensitive strains belonging to five different STEC serogroups (O26, O103, O104, O111 and O157; n=10) were used in this study. All strains were adapted in PJ (pH 3·8) stored at 4 and 20°C for 24h, and then the relative transcription levels of genes in all strains were quantified using a real-time quantitative-PCR assay. After adaptation in PJ, the STEC strains were exposed to SGF (pH 1·5 and 2·0) at 37°C for 2h. Generally, the STEC adapted in PJ at 4°C displayed enhanced survival compared to acid adaptation in PJ at 20°C and nonadapted controls with exposure to SGF (P<0·05). Moreover, resistant strains exhibited higher survival rates compared to sensitive strains (P<0·05). Overall, adaptation at 4°C resulted in significantly (P<0·05) enhanced gene expression levels in PJ, and transcript levels of gadA were higher than those of the rpoS and adiA genes. The up-regulation of AR genes due to adaptation in PJ at low temperature may increase STEC survival in acidic environments such as the gastrointestinal tract. Some non-O157 STEC strains, including serotypes O103:H2 and O111:H8, showed relatively high AR levels similar to those of STEC O157:H7. Induction of AR genes in acidic fruit juice, and potentially in other acidic foods may increase the risk of foodborne illness by non-O157 STEC serogroups.

Chitosan Microparticles Exert Broad-Spectrum Antimicrobial Activity against Antibiotic-Resistant Micro-organisms without Increasing Resistance.

Antibiotic resistance is growing exponentially, increasing public health concerns for humans and animals. In the current study, we investigated the antimicrobial features of chitosan microparticles (CM), engineered from chitosan by ion gelation, seeking potential application for treating infectious disease caused by multidrug resistant microorganisms. CM showed excellent antimicrobial activity against a wide range of microorganisms, including clinically important antibiotic-resistant pathogens without raising resistant mutants in serial passage assays over a period of 15 days, which is a significantly long passage compared to tested antibiotics used in human and veterinary medicine. In addition, CM treatment did not cause cross-resistance, which is frequently observed with other antibiotics and triggers multidrug resistance. Furthermore, CM activity was examined in simulated gastrointestinal fluids that CM encounter when orally administered. Antimicrobial activity of CM was exceptionally strong to eliminate pathogens completely. CM at a concentration of 0.1 μg/mL killed E. coli O157:H7 (5 × 10(8) CFU/mL) completely in synthetic gastric fluid within 20 min. Risk assessment of CM, in an in vitro animal model, revealed that CM did not disrupt the digestibility, pH or total volatile fatty acid production, indicating that CM likely do not affect the functionality of the rumen. Given all the advantages, CM can serve as a great candidate to treat infectious disease, especially those caused by antibiotic-resistant pathogens without adverse side effects.

Evaluation of the bio-accessible non-process element concentrations in slaker grits by synthetic sweat and gastric fluids extraction

Aim: The non-process elements (NPEs) or more correctly inorganic process contaminants are compounds that are not necessary for the chemistry of kraft pulping, e.g. heavy metals. The aim of this study was to determine the extractability of NPEs in the slaker grits using artificial sweat and gastric fluids to assess the potential occupational risk from grits handling. Methods: The slaker grits investigated in this study were sampled in November 2013 from the outlet of the causticizing process at a pulp mill located in Finland. The bio-accessible concentrations of NPEs in the slaker grits were determined using their solubility (extractability) in synthetic sweat (pH = 6.4) and gastric (pH = 1.5) fluids. Results: The highest bio-accessible concentration of non-process elements (NPEs) in the synthetic sweat fluid was observed for Al (14.0 mg/kg; d.w.), and in the synthetic gastric fluid for Al (4920 mg/kg; d.w.) and Ba (163 mg/kg; d.w.). If we disregard the NPEs whose concentrations were lower than the detection limits, the synthetic gastric fluid was able to release between 42.6% (Pb) and 81.8% (Ni) of the total element concentrations of NPEs in the slaker grits. Conclusion: We conclude that due to the very high bio-accessible concentration of certain NPEs in the synthetic gastric fluid, it is necessary to prevent the ingestion and penetration of slaker grits particles across the human gastrointestinal tract.

Survival of Listeria monocytogenes during Production and ripening of traditional Turkish Savak tulum cheese and in synthetic gastric fluid.

This study investigated the survival and acid tolerance of Listeria monocytogenes during the 2-day processing stage and 90-day ripening of Savak tulum cheese, a traditional cheese in Turkey. Experimental Savak tulum cheese was produced from raw sheep's milk that was inoculated with a L. monocytogenes mixture consisting of five strains (average 7.0 log CFU/ml) and was ripened at 6°C for 90 days. Microbiological and chemical analyses and acid exposure experiments in synthetic gastric fluid (SGF) (pH 1.5 to 2.5) were carried out on days 1 and 2 during processing and on days 0, 15, 30, 45, 60, and 90 during ripening. The numbers of L. monocytogenes did not decrease during processing, but a total of 4.1 log CFU/g reduction was observed during ripening. Throughout the ripening period, L. monocytogenes cells survived direct 90-min exposures of the cheese samples to SGF. These results suggest that, although the pathogen numbers decreased in Savak tulum cheese ripened at 6°C for 90 days, a sublethal environment may have occurred in the cheese during the production stage, activating the acid-tolerance mechanisms of the pathogen and allowing L. monocytogenes to maintain its viability in the SGF for 90 min.

Effect of Selected Types of Beer on Bacteria of the Genus Arcobacter

The genus Arcobacter is related to the well-known human pathogen, Campylobacter jejuni, and has been linked to human diseases. In this study, the survival of Arcobacter spp. in various concentrations of ethanol, in various samples of beers, and in a model stomach has been investigated. For most of these bacteria, a concentration of 10 % ethanol was determined to be the minimum inhibitory concentration. The fact that these organisms are able to survive under these conditions may have an impact in the food processing industry. We studied the activity of beer against arcobacters. These bacteria were killed in all samples of beer within 30 min. A model stomach, containing a food matrix and a synthetic gastric fluid, was used to deduce the effect of beer against Arcobacter spp. during food consumption. Complete inactivation of all monitored arcobacters was detected usually within 15 min. However, the presence of beer does not potentiate the effect of gastric fluid against these bacteria. This is apparently the first study focusing upon the effect of beer on Arcobacter spp.

The antimicrobial effect of wine on Bacillus cereus in simulated gastro-intestinal conditions

This study aimed to evaluate the antimicrobial activity of wine against Bacillus cereus vegetative cells and spores. The results clearly show that wine exerts a strong inactivation effect against vegetative cells of B. cereus. The red wine tested inactivated stationary phase cultures to undetectable numbers in less than 10 s. Thus, further inactivation assays were carried out with wine diluted with water (1:4 and 1:8). Diluted wine 1:4 caused a reduction of approximately 5 log cycles on viable cell counts in 20 s. On the other hand, B. cereus spores were found to be highly resistant to wine exposure. The influence of wine components (organic acids, ethanol and phenolic compounds) was investigated on vegetative cells. The wine organic acids tested exhibited a strong inactivation effect, and, when combined with ethanol, a slight synergistic effect was observed. The wine phenolic compounds assayed displayed no activity against the vegetative cells at the concentrations tested. At the simulated gastric conditions studied (in the presence of food), wine diminished considerably the number of B. cereus viable cells in addition to the effect of the synthetic gastric fluid. The behaviour of B. cereus spores under gastro-intestinal conditions was also evaluated. In a consumption-like scenario, the addition of wine led to lower total counts (vegetative cells + spores) of B. cereus in the simulated intestine conditions, showing that wine inhibits the proliferation of the vegetative cells obtained from the germination of spores. This work provides evidence that consumption of wine during a meal may diminish the number of viable cells of B. cereus and reduces the impact of the germination of spores that may occur in the small intestine, thus lowering the risk of toxi-infection that may be caused by this pathogen.

Purine Biosynthesis Mutants (purAandpurB) of Serotype 4bListeria monocytogenesAre Severely Attenuated for Systemic Infection in Intragastrically Inoculated A/J Mice

In this study, we demonstrate that purA and purB transposon mutants of serotype 4b Listeria monocytogenes were severely impaired in their ability to colonize the gastrointestinal tract and cause systemic infection of the spleen, liver, and gallbladder following intragastric inoculation of A/J mice. The mutant strains were also impaired in their ability to multiply within Caco-2 human intestinal epithelial cells. Neither mutant was affected in resistance to synthetic gastric fluid (pH 4.5). These findings indicate that purine biosynthesis is critical for gastrointestinal virulence of L. monocytogenes serotype 4b in mice.

Removal of zearalenone toxin from synthetics gastric and body fluids using talc and diatomite: A batch kinetic study

Adsorption kinetics of zearalenone (ZEA) toxin from synthetic gastric fluid (SGF) and synthetic body fluid (SBF) by talc and diatomite was studied in the batch experiments. Chemical composition, morphology and structure of the used adsorbents were examined by scanning electron microscopy, FTIR spectroscopy and low-temperature nitrogen adsorption/desorption method. High performance liquid chromatography (HPLC) method was used for ZEA determining. The study results showed that ZEA is more effectively adsorbed on the talc (73% and 54% from SGF and SBF respectively). The efficiency on the diatomite was lower (53% and 42% from SGF and SBF respectively). The first order kinetics model was applied to describe the adsorption process. Rate of the ZEA adsorption from SGF is very rapid initially with about 95% of amount of the toxin adsorbed during first 5min, while ZEA is adsorbed from SBF in two steps. The values of determined Gibbs free energy of adsorption (from −13 to −17kJ/mol) indicated that adsorption of ZEA toxin by the both adsorbents are spontaneous and exothermic.

Differential Gene Expression of E. coli O157:H7 in Ground Beef Extract Compared to Tryptic Soy Broth

E. coli O157:H7 is an important foodborne pathogen, and ground beef is a common vehicle of infection. DNA microarrays have been used for transcriptomic studies of E. coli O157:H7 using laboratory media; however, analysis of gene expression in complex matrices such as food are lacking. This study compared gene expression profiles of E. coli O157:H7 Sakai strain in raw ground beef extract (GBE) and tryptic soy broth (TSB). Total RNA was isolated from GBE and TSB after 2 h of incubation with E. coli O157:H7. Following reverse transcription (RT) of the RNA, labeled cDNA was hybridized to microarrays representing 9608 open reading frames (Operon; Genome Array-Ready Oligo Set) corresponding to 4 genomes of E. coli strains and 3 plasmids. There were 74 up-regulated (genes involved in protein and polysaccharide biosynthesis, transcription factors, membrane transport proteins, and acid shock proteins) and 54 down-regulated (encoding proteins for energy metabolism, biosynthesis of cofactors, transporters of small molecules, and transcription factors and enzymes responsible for protein degradation) genes in E. coli O157:H7 grown in GBE compared to TSB, respectively. Furthermore, compared to incubation in TSB, E. coli O157:H7 incubated in GBE for 2 h showed significantly increased survival when exposed to synthetic gastric fluid, pH 1.5. This study demonstrated that microarray analyses can be performed using complex food matrices, and gene expression of E. coli O157:H7 differs in TSB compared to GBE. The information will be useful for identification of genes that can be employed as potential targets for interventions to control E. coli O157:H7.

Phenotypic diversity of Escherichia coli O157:H7 strains associated with the plasmid O157.

Escherichia coli O157:H7, a food-borne pathogen, causes hemorrhagic colitis and the hemolytic-uremic syndrome. A putative virulence factor of E. coli O157:H7 is a 60-MDa plasmid (pO157) found in 99% of all clinical isolates and many bovine-derived strains. The well characterized E. coli O157:H7 Sakai strain (Sakai) and its pO157-cured derivative (Sakai-Cu) were compared for phenotypic differences. Sakai-Cu had enhanced survival in synthetic gastric fluid, did not colonize cattle as well as wild-type Sakai, and had unchanged growth rates and tolerance to salt and heat. These results are consistent with our previous findings with another E. coli O157:H7 disease outbreak isolate ATCC 43894 and its pO157-cured (43894-Cu). However, despite the essentially sequence identical pO157 in these strains, Sakai-Cu had changes in antibiotic susceptibility and motility that did not occur in the 43894-Cu strain. This unexpected result was systematically analyzed using phenotypic microarrays testing 1,920 conditions with Sakai, 43894, and the plasmid-cured mutants. The influence of the pO157 differed between strains on a wide number of growth/survival conditions. Relative expression of genes related to acid resistance (gadA, gadX, and rpoS) and flagella production (fliC and flhD) were tested using quantitative real-time PCR and gadA and rpoS expression differed between Sakai-Cu and 43894-Cu. The strain-specific differences in phenotype that resulted from the loss of essentially DNA-sequence identical pO157 were likely due to the chromosomal genetic diversity between strains. The O157:H7 serotype diversity was further highlighted by phenotypic microarray comparisons of the two outbreak strains with a genotype 6 bovine E. coli O157:H7 isolate, rarely associated with human disease.

Hazardous chemicals in synthetic turf materials and their bioaccessibility in digestive fluids

Many synthetic turf fields consist of not only artificial grass but also rubber granules that are used as infill. The public concerns about toxic chemicals possibly contained in either artificial (polyethylene) grass fibers or rubber granules have been escalating but are based on very limited information available to date. The aim of this research was to obtain data that will help assess potential health risks associated with chemical exposure. In this small-scale study, we collected seven samples of rubber granules and one sample of artificial grass fiber from synthetic turf fields at different ages of the fields. We analyzed these samples to determine the contents (maximum concentrations) of polycyclic aromatic hydrocarbons (PAHs) and several metals (Zn, Cr, As, Cd, and Pb). We also analyzed these samples to determine their bioaccessible fractions of PAHs and metals in synthetic digestive fluids including saliva, gastric fluid, and intestinal fluid through a laboratory simulation technique. Our findings include: (1) rubber granules often, especially when the synthetic turf fields were newer, contained PAHs at levels above health-based soil standards. The levels of PAHs generally appear to decline as the field ages. However, the decay trend may be complicated by adding new rubber granules to compensate for the loss of the material. (2) PAHs contained in rubber granules had zero or near-zero bioaccessibility in the synthetic digestive fluids. (3) The zinc contents were found to far exceed the soil limit. (4) Except one sample with a moderate lead content of 53 p.p.m., the other samples had relatively low concentrations of lead (3.12-5.76 p.p.m.), according to soil standards. However, 24.7-44.2% of the lead in the rubber granules was bioaccessible in the synthetic gastric fluid. (5) The artificial grass fiber sample showed a chromium content of 3.93 p.p.m., and 34.6% and 54.0% bioaccessibility of lead in the synthetic gastric and intestinal fluids, respectively.

Characterization of Virulence Properties of Listeria monocytogenes Serotype 4b Strains of Different Origins

In this study, the virulence heterogeneity of Listeria monocytogenes serotype 4b strains of different origins was analysed on different levels. On one hand, the survival of L. monocytogenes strains in synthetic gastric fluid was studied. On the other hand, the pathogenic potential of strains with different inlB expression levels was analysed in an A/J mouse model for gastrointestinal listeriosis. Differences in survival capacity in gastric fluid and in in vivo virulence potential were observed between the tested strains. No clear correlation between the origin and the obtained data could be made. However, these results confirm the existence of heterogeneity in virulence potential of L. monocytogenes serotype 4b strains.