Joint effect of heat, pH and grape extract on Bacillus cereus spores survival in a rice solution.

Rice due to its high carbohydrate content, is an ideal medium for Bacillus cereus growth, a spore-producing microorganism. The objective of this study was to determine the antimicrobial activity of a grape extract in combination with heat treatments and different pH against B. cereus spores in a rice solution. The survivor data obtained were fitted to the Weibull survival function, and the values of parameters a and b (scale and shape indexes, respectively) were determined. Results showed that the grape extract affected the survival of B. cereus spores at 90 °C and 95 °C, reaching greater logarithmic reductions in acidic pH values. This behaviour was reflected in a parameter of the Weibull survival function which decreased as the temperature increased and at acidic pH values. In addition, a secondary model was developed by relating the logarithm of a to the independent variables (temperature and pH). A global model relating B. cereus inactivation with temperature and pH was developed, and validated by calculating the accuracy factor. The results demonstrate the usefulness of grape extract as a by-product, which can be used as an additional control measure for rice, especially when combined with mild heat treatments and acidic pH values.

Joint effect of temperature and insect chitosan on the heat resistance of Bacillus cereus spores in rice derivatives.

The heat resistance of Bacillus cereus spores inoculated in a rice substrate supplemented with insect chitosan as an alternative antimicrobial was studied. Two concentrations of insect chitosan were considered in order to assess the role of the insect chitosan concentration during the heat process. Results of the study indicated that the DT values were higher in the substrate without chitosan than in the substrate containing chitosan thus indicating a greater heat resistance to heat treatment of the microorganism inoculated in the substrate without chitosan. This behaviour was also evidenced in the survival curves. There were no great differences between either of the insect chitosan concentrations tested regarding the DT values. The z values were 9.8°C on rice substrate and8.9°C on rice substrate supplemented with insect chitosan at 150 µg/mL and 10.7°C on rice substrate supplemented with 250 µg/mL of insect chitosan. The chitosan concentration appears to affect the z value of the microorganism. Our results indicate that the combination of heat with insect chitosan as an antimicrobial on foodstuffs subjected to cooking is feasible and can improve the safety of rice derivatives.

Insect chitosan as a natural antimicrobial against vegetative cells of Bacillus cereus in a cooked rice matrix.

This study investigates the antimicrobial activity of insect chitosan against vegetative cells of Bacillus cereus in a rice matrix. Sample culture solutions were prepared with different concentrations of insect chitosan (150, 180, 220 and 250 µg/mL) and tested at three temperatures (30 °C, 20 °C and 10 °C), which simulate different storage temperature scenarios of precooked rice. The results indicate that insect chitosan has antimicrobial activity that depends on temperature and chitosan concentration. For the assays with chitosan at 10 °C, all concentrations were bactericidal during the study time, reaching a maximum inactivation of 6 log cycles for 250 µg/mL. At 20 °C and at 30 °C a bacteriostatic activity was observed for concentrations of 150 µg/mL and 180 µg/mL. Results also showed that concentrations of 220 µg/mL and 250 µg/mL were bactericidal for all the temperatures tested during the storage time. When rice is cooked and not stored at an appropriate temperature, below 10 °C, the consumer's health is at risk. In these cases, insect chitosan could be a good additional control measure to control B. cereus growth and toxin formation in cooked rice.

Surface Micro Discharge-Cold Atmospheric Pressure Plasma Processing of Common House Cricket Acheta domesticus Powder: Antimicrobial Potential and Lipid-Quality Preservation.

The growing world population and the need to reduce the environmental impact of food production drive the exploration of novel protein sources. Insects are being cultivated, harvested, and processed to be applied in animal and human nutrition. The inherent microbial contamination of insect matrices requires risk management and decontamination strategies. Thermal sterilization results in unfavorable cooking effects and oxidation of fatty acids. The present study demonstrates the risk management in Acheta domesticus (home cricket) powder with a low-energy (8.7-22.0 mW/cm2, 5 min) semi-direct surface micro discharge (SMD)-cold atmospheric pressure plasma (CAPP). At a plasma power density lower than 22 mW/cm2, no degradation of triglycerides (TG) or increased free fatty acids (FFA) content was detected. For mesophilic bacteria, 1.6 ± 0.1 log10 reductions were achieved, and for Enterobacteriaceae, there were close to 1.9 ± 0.2 log10 reductions in a layer of powder. Colonies of Bacillus cereus, Bacillus subtilis, and Bacillus megaterium were identified via the mass spectral fingerprint analyzed with matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS). The spores of these Bacillus strains resisted to a plasma power density of 22 mW/cm2. Additional inactivation effects at non-thermal, practically non-oxidative conditions are supposed for low-intensity plasma treatments combined with the powder's fluidization.

Benefits and Challenges in the Incorporation of Insects in Food Products.

Edible insects are being accepted by a growing number of consumers in recent years not only as a snack but also as a side dish or an ingredient to produce other foods. Most of the edible insects belong to one of these groups of insects such as caterpillars, butterflies, moths, wasps, beetles, crickets, grasshoppers, bees, and ants. Insect properties are analyzed and reported in the articles reviewed here, and one common feature is nutrimental content, which is one of the most important characteristics mentioned, especially proteins, lipids, fiber, and minerals. On the other hand, insects can be used as a substitute for flour of cereals for the enrichment of snacks because of their high content of proteins, lipids, and fiber. Technological properties are not altered when these insects-derived ingredients are added and sensorial analysis is satisfactory, and only in some cases, change in color takes place. Insects can be used as substitute ingredients in meat products; the products obtained have higher mineral content than traditional ones, and some texture properties (like elasticity) can be improved. In extruded products, insects are an alternative source of proteins to feed livestock, showing desirable characteristics. Isolates of proteins of insects have demonstrated bioactive activity, and these can be used to improve food formulations. Bioactive compounds, as antioxidant agents, insulin regulators, and anti-inflammatory peptides, are high-value products that can be obtained from insects. Fatty acids that play a significant role in human health and lipids from insects have showed positive impacts on coronary disease, inflammation, and cancer. Insects can be a vector for foodborne microbial contamination, but the application of good manufacturing practices and effective preservation techniques jointly with the development of appropriate safety regulations will decrease the appearance of such risks. However, allergens presented in some insects are a hazard that must be analyzed and taken into account. Despite all the favorable health-promoting characteristics present in insects and insects-derived ingredients, willingness to consume them has yet to be generalized.

Applications of Natural Antimicrobials in Food Packaging and Preservation.

In the food science field, the term "antimicrobial" basically refers to active substances of synthetic or natural origin, that are directly or indirectly present in a specific food, packaging material or food contact surface that affect the viability or the growth of microorganisms in that matrix [...].

Effect of HHP, Enzymes and Gelatin on Physicochemical Factors of Gels Made by Using Protein Isolated from Common Cricket (Acheta domesticus).

The effect of high hydrostatic pressure (HHP) combined with enzymatic methods or gelatin incorporation in the gelation process of protein isolated from Acheta domesticus was investigated. The results indicate that transglutaminase (TGasa) or glucose oxidase (GOx) enzymes can induce reversible aggregation in dispersions of insoluble protein fractions and increase viscosity in dispersions of soluble fractions, but does not induce gel formation even after HHP treatment; in consequence, enzymatic treatment on cricket protein can be used to increase viscosity but not to form gels. It is technically feasible to obtain gels by adding 2% porcine gelatin to dispersions of protein fractions and subjecting them to HHP. The firmness and syneresis variation values of those gels during storage depended on the protein extracted fraction (insoluble or soluble protein) and on the concentration of protein used. The highest hardness and lowest syneresis was found with the gels obtained from the insoluble fraction at 11 and 15% (w/w) protein concentration. Color difference (ΔE* > 3) appreciable to the naked eye was observed along the storage period and no noticeable pH variations were found after 28 days of storage. Results indicate that new raw materials of interest can be developed for the food industry based on cricket protein isolates, to make high-protein foods which could be applied in a wide variety of different food applications including 3D printing or fat substitution.

Risk of Bacillus cereus in Relation to Rice and Derivatives.

Rice is a very popular food throughout the world and the basis of the diet of the citizens of many countries. It is used as a raw material for the preparation of many complex dishes in which different ingredients are involved. Rice, as a consequence of their cultivation, harvesting, and handling, is often contaminated with spores of Bacillus cereus, a ubiquitous microorganism found mainly in the soil. B. cereus can multiply under temperature conditions as low as 4 °C in foods that contain rice and have been cooked or subjected to treatments that do not produce commercial sterility. B. cereus produces diarrhoeal or emetic foodborne toxin when the consumer eats food in which a sufficient number of cells have grown. These circumstances mean that every year many outbreaks of intoxication or intestinal problems related to this microorganism are reported. This work is a review from the perspective of risk assessment of the risk posed by B. cereus to the health of consumers and of some control measures that can be used to mitigate such a risk.

In vivo Assessment of Cold Atmospheric Pressure Plasma Technology on the Bioactivity of Spirulina.

The present study challenges the in vivo assessment of cold atmospheric pressure plasma (CAPP) technology on the bioactive activity (antioxidant/antiaging and antimicrobial potential) of Spirulina powder, using Caenorhabditis elegans as an animal model. Surface microdischarge cold atmospheric pressure plasma (SMD-CAPP) treatment was 3.3 W discharge power for 7 min. C. elegans lifespan and egg laying were used as indicators of antioxidant/antiaging potential of Spirulina (1 mg/mL), when grown with Spirulina CP-treated [E_SCP] and untreated [E_S], compared with a control [E_0] (non-supplemented with Spirulina). According to our results, under both Spirulina supplemented media [E_SCP and E_S] and for the first 17 days, nematodes experienced an increase in lifespan but without significant differences (p > 0.05) between control and Spirulina CP-treated. Regarding the in vivo assay of the antimicrobial potential of Spirulina against Salmonella enterica serovar Typhimurium (infected worms), no significant differences (p > 0.05) were found between the three exposure scenarios (control [S_0]; Spirulina supplemented media [S_S]; CP-treated Spirulina supplemented media [S_SCP]). According to present results, CAPP-treatment do not influence negatively the lifespan of C. elegans but a reduction in the Spirulina antiaging potential was found. No in vivo modifications in antimicrobial activity seem to be linked to CAPP-processed Spirulina.

Rare non-papillary lithiasis of calcium oxalate monohydrate generated on a central core of potassium urate.

This report describes a patient who developed a spheroidal calculus with a central part composed of potassium urate, surrounded by a continuous layer of calcium oxalate monohydrate with crystals of calcium oxalate dihydrate on the surface. The mechanism of calculus development is also suggested.

Antimicrobial effect of insect chitosan on Salmonella Typhimurium, Escherichia coli O157:H7 and Listeria monocytogenes survival.

The antimicrobial capability of chitosan from Tenebrio molitor as compared with chitosan from crustacean (Penaeus monodon) on different pathogenic microorganisms of concern in food safety was studied. The antimicrobial effect was tested at pH 5 and pH 6.2 and at two different initial concentrations (103 or 106 CFU/mL). Results indicated that chitosan from both sources have antimicrobial activity, although the effect depended on the microorganism considered (Salmonella Typhimurium, Listeria monocytogenes and Escherichia coli O157:H7). Our results indicated that Salmonella was the most resistant bacteria, and that chitosan from insect was less active than chitosan from crustacean, especially against Salmonella. Another important factor on antimicrobial activity was the pH of the sample. When chitosan was added to a solution with a pH of 6.2 it was more active against Listeria and Escherichia coli, than at pH 5.00. Besides, the effect of chitosan appears to decrease with the incubation time, since some increases in counts were observed on E. coli and Salmonella after the 24 and 49 hours of incubation.

Caenorhabditis elegans as an in vivo model to assess fucoidan bioactivity preventing Helicobacter pylori infection.

Currently, Helicobacter pylori is the unique biological carcinogenic agent. The search for antimicrobial alternatives to antibiotics against this pathogen has been categorized as a priority due to the drastic failure associated with current applied antibiotic therapy. The present study assessed the bioactive antimicrobial capability of fucoidan ("Generally Recognized as Safe" approval - European Commission December 2017) from different species of Phaeophyceae algae (Fucus vesiculosus, Undaria pinnatifida, Macrocystis pyrifera) against H. pylori. All the studied fucoidans showed bacteriostatic and bactericidal effects at the studied concentrations [5-100] µg ml-1 and exposure times [0-7 days]. The most effective anti-H. pylori fucoidan was validated in Caenorhabditis elegans as an in vivo model. C. elegans feed was supplemented with Undaria pinnatifida [0-100] µg ml-1 fucoidan, resulting in a significant improvement in lifespan, lowered H. pylori concentration in the digestive tract, and increased egg-laying pattern. New research lines proposing this compound as an active agent in nutraceutical and preventive novel therapies should be opened.

Analysis of urine composition from split 24-h samples: use of 12-h overnight samples to evaluate risk factors for calcium stones in healthy and stone-forming children.

INTRODUCTION: The analysis of 24-h urine is the gold standard to diagnose metabolic abnormalities in the stone-forming patient. However, urinary composition changes throughout the day and analyzing the whole 24-h urine may mask peaks of increased risk of crystallization. OBJECTIVE: To examine variations of stone-promoting and stone-inhibiting factors in urine using split 24-h samples from healthy and stone-forming children. STUDY DESIGN: Urine was collected from 87 healthy and 26 stone-forming children using a split collection procedure (12-h daytime urine and 12-h overnight urine). Urine volume, pH, calcium (Ca), magnesium (Mg), phosphate (P), citrate (Cit), uric acid (Ur), and oxalate (Ox) were determined, and the Ca/Cit ratio was calculated. RESULTS: The overnight urine samples in both groups had higher levels of P and Mg, lower volume, lower pH, and less citrate and uric acid. As can be seen in the table, higher percentages of healthy and stone-forming children had altered 12-h night urine than 24-h urine with regards to Ca/Cr, Cit/Cr and Ca/Cit ratios. All healthy subjects and all stone-forming children (except one) with altered Cit/Cr ratios or Ca/Cit ratios in the 24-h sample also had altered ratios in the 12-h overnight sample. DISCUSSION: This study indicates that urine composition changes throughout the day, and that there is daily variability in most of the parameters related to kidney stone formation. Furthermore, 12-h overnight samples seem to be more sensitive than 24-h samples in detecting the most common urinary abnormalities. The main limitation of this study is the relative low sample size of stone-forming children, owing to the low prevalence of nephrolithiasis in childhood. CONCLUSIONS: We observed a higher excretion of stone-promoting substances and a lower citrate in urine at night. However, the study results do not provide enough evidence to conclude that the use of a 12-h overnight sample collection can replace 24 h urine analysis in the metabolic evaluation of children with lithiasis.

Optimizing High Pressure Processing Parameters to Produce Milkshakes Using Chokeberry Pomace.

High hydrostatic pressure is a non-thermal treatment of great interest because of its importance for producing food with additional or enhanced benefits above their nutritional value. In the present study, the effect of high hydrostatic pressure processing parameters (200-500 MPa; 1-10 min) is investigated through response surface methodology (RSM) to optimize the treatment conditions, maximizing the phenol content and antioxidant capacity while minimizing microbiological survival, in milkshakes prepared with chokeberry pomace (2.5-10%). The measurement of fluorescence intensity of the samples was used as an indicator of total phenolic content and antioxidant capacity. The results showed that the intensity of the treatments had different effects on the milkshakes. The RSM described that the greatest retention of phenolic compounds and antioxidant capacity with minimum microbiological survival were found at 500 MPa for 10 min and 10% (w/v) chokeberry pomace. Therefore, this study offers the opportunity to develop microbiologically safe novel dairy products of high nutritional quality.

Occurrence of Salmonella typhimurium resistance under sublethal/repeated exposure to cauliflower infusion and infection effects on Caernohabditis elegans host test organism.

Resistant bacteria to antimicrobials are increasingly emerging in medical, food industry and livestock environments. The present research work assesses the capability of Salmonella enterica var Typhimurium to become adapted under the exposure to a natural cauliflower antimicrobial by-product infusion in consecutive repeated exposure cycles. Caenorhabditis elegans was proposed as in vivo host-test organism to compare possible changes in the virulent pattern of the different rounds treated S. enterica var Typhimurium and untreated bacterial cells. According to the obtained results, S. enterica var Typhimurium was able to generate resistance against a repeated exposure to cauliflower by-product infusion 5% (w/v), increasing the resistance with the number of exposed repetitions. Meanwhile, at the first exposure, cauliflower by-product infusion was effective in reducing S. enterica var Typhimurium (≈1 log10 cycle), and S. enterica var Typhimurium became resistant to this natural antimicrobial after the second and third treatment-round and was able to grow (≈1 log10 cycle). In spite of the increased resistance observed for repeatedly treated bacteria, the present study reveals no changes on C. elegans infection effects between resistant and untreated S. enterica var Typhimurium, according to phenotypic parameters evaluation (lifespan duration and egg-laying).

Effect of the addition of liquid whey from cheese making factory on the physicochemical properties of whey protein isolate gels made by high hydrostatic pressure.

The objective of the present work was to study the effect of liquid whey from the cheese production process on the gels developed by high hydrostatic pressure from whey protein isolate powder (WPI). Changes in pH, color, textural parameters, and water retention capacity of the gels obtained were analyzed during storage for 28 days at refrigeration temperature (4 °C). Mixtures of liquid whey from cheese making processes and different WPI percentages gave gels with different characteristics after being processed by high hydrostatic pressures. The pH values and color parameters (L*, a*, b*) varied slightly, depending directly on WPI concentration and storage time. The values of hardness, elasticity, and cohesiveness were dependent on the liquid medium used to dissolve the WPI (liquid cheese whey or distilled water), WPI concentration, and storage time. The use of liquid cheese whey for gel formation favored water retention, reducing the appearance of syneresis (exudation). The results obtained in the present study indicated a possible use and revalorization of cheese whey obtained in cheese production to obtain WPI gels with improved physicochemical properties, using high hydrostatic pressure as technology for their production.

Effect of sample time on urinary lithogenic risk indexes in healthy and stone-forming adults and children.

BACKGROUND: The diagnosis and follow-up of stone forming patients is usually performed by analysis of 24-h urine samples. However, crystallization risk varies throughout the day, being higher at night. The main objective of this study is to evaluate the urinary crystallization risk in adults and children by calculating risk indexes based on different collection periods. METHODS: The study included 149 adults (82 healthy and 67 stone-formers) and 108 children (87 healthy and 21 stone-formers). 24-h urine was collected, divided into 12-h daytime sample (8 am to 8 pm), and 12-h overnight sample (8 pm to 8 am next morning). Solute concentrations, the calcium to citrate ratio (Ca/Cit), and the ion activity product of calcium oxalate (AP[CaOx]) and calcium phosphate (AP[CaP]) were calculated in each 12-h sample and in overall 24-h urine. Assessments were also related to stone type. RESULTS: Ca/Cit and AP(CaOx) were significantly higher in stone forming patients than in healthy subjects. The 12-h overnight samples had the highest values for both risk indexes, confirming a greater risk for crystallization at night. The AP(CaP) index was significantly higher in patients with pure hydroxyapatite stones than healthy controls, but was not significantly different between stone-formers overall and healthy controls. CONCLUSIONS: The calculation of risk indexes is a simple method that clinicians can use to estimate crystallization risk. For this purpose, the use of 12-h overnight urine may be a reliable alternative to 24-h collections.

Thermal stabilization of probiotics by adsorption onto porous starches.

Industrial processing factors, such as temperature, compromise the viability of probiotic cells. Objective was to develop a system to thermally stabilize probiotic bacteria based on porous starches and using biopolymers as coating materials (gelatinized starch, guar gum and xanthan gum). Porous starches from corn and rice starches, having controlled number and size of porous were used as supporting material. Scanning electron microscopy confirmed the adsorption of the microorganism, leading microcapsules with corn starch but aggregates with rice starch. Surface pores of rice starch increased the encapsulation yield of rice starch around 10%, but that effect was not observed in porous corn starch. The highest encapsulation yield was obtained with porous starches coated with gelatinized starch, which ranged from 92 to 100%. Microencapsulates made with porous starches with small pores, like the ones obtained with α-amylase, and coated with gelatinized starch resulted in the highest thermal resistance at 55 °C.

Use of high hydrostatic pressure to inactivate natural contaminating microorganisms and inoculated E. coli O157:H7 on Hermetia illucens larvae.

A chemical and microbiological characterization on Hermetia illucens larvae was carried out as well as an inactivation study of natural contaminating microorganisms and inoculated E. coli O157:H7 in black soldier larvae by using High Hydrostatic Pressure (250 to 400 MPa, for 1.5 to 15 min). Hermetia illucens was mainly composed of proteins (46.49%, d.m.) followed by fat (37.88%, d.m.). Larvae had a high contamination load of Total Aerobic Mesophilic bacteria (AMB) (1.58x107 cfu/g) and Enterobacteriaceae (1.15x106cfu/g). The presence of pathogenic microorganism varied: no Listeria spp. were found, but Salmonella (1.15x106 cfu/g) and E. coli (7.08x105 cfu/g) were detected in the larvae extract. High Hydrostatic Pressure (HHP) was effective against natural contaminating yeasts and molds producing more than 5 log cycle reductions at 400 MPa for any of the times considered (2.5 to 7 min), but a low reduction of total microbial load was achieved. The inactivation level of larvae inoculated with E. coli O157:H7 varied. At 400 MPa for 7 min more than 5 log cycle reductions were achieved. Among the three inactivation models studied, the one that best described the inactivation pattern of the cells, according to the Akaike index, was the Biphasic model.

S. Typhimurium virulence changes caused by exposure to different non-thermal preservation treatments using C. elegans.

The aims of this research study were: (i) to postulate Caenorhabditis elegans (C. elegans) as a useful organism to describe infection by Salmonella enterica serovar Typhimurium (S. Typhimurium), and (ii) to evaluate changes in virulence of S. Typhimurium when subjected repetitively to different antimicrobial treatments. Specifically, cauliflower by-product infusion, High Hydrostatic Pressure (HHP), and Pulsed Electric Fields (PEF). This study was carried out by feeding C. elegans with different microbial populations: E. coli OP50 (optimal conditions), untreated S. Typhimurium, S. Typhimurium treated once and three times with cauliflower by-product infusion, S. Typhimurium treated once and four times with HHP and S. Typhimurium treated once and four times with PEF. Bayesian survival analysis was applied to estimate C. elegans lifespan when fed with the different microbial populations considered. Results showed that C. elegans is a useful organism to describe infection by S. Typhimurium because its lifespan was reduced when it was infected. In addition, the application of antimicrobial treatments repetitively generated different responses: when cauliflower by-product infusion and PEF treatment were applied repetitively the virulence of S. Typhimurium was lower than when the treatment was applied once. In contrast, when HHP treatment was applied repetitively, the virulence of S. Typhimurium was higher than when it was applied once. Nevertheless, in all the populations analyzed treated S. Typhimurium had lower virulence than untreated S. Typhimurium.