High Digestion Efficiency Research Articles

Microplastic extraction from digestive tracts of large decapods

The reliable quantification of microplastic contamination in chitinous organisms requires validated methods to remove interfering complex organic and inorganic material. This study trialled KOH, H2O2 and HNO3 digestion methods on the digestive tracts of two large decapods (Panulirus cygnus and Portunus armatus) to validate a protocol that facilitates reliable microplastic extraction. KOH digestion provided the best recovery (>95 %) of all polymers (e.g. polyamide, polyethylene, polyethylene terephthalate, polypropylene, polystyrene and polyvinyl chloride), with the lowest impact to their physical morphology and chemical spectra. While HNO3, and HNO3 + H2O2 treatments were more effective at digesting chitin, they destroyed polyamide, and altered several other polymers. High digestion efficiency did not result in high matrix clarification or high microplastic recovery for large decapods. This study emphasises the importance of validating species-specific microplastic extraction methods, whilst proposing additional post-digestion protocols, such as density separation, for complex samples, that can be applied in future research investigating plastic contamination in large decapods.

Dietary protein from different sources escapes host digestion and is differentially modified by the microbiota.

Protein is an essential macronutrient and variations in its source and quantity have been shown to impact long-term health outcomes. Differential health impacts of dietary proteins from various sources are likely driven by differences in their digestibility by the host and subsequent availability to the intestinal microbiota. However, our current understanding regarding the fate of dietary proteins from different sources in the gut, specifically how component proteins within these sources interact with the host and the gut microbiota, is limited. To determine which dietary proteins are efficiently digested by the host and which proteins escape host digestion and are used by the gut microbiota, we used high-resolution mass spectrometry to quantify the proteins that make up different dietary protein sources before and after digestion in germ-free and conventionally raised mice. Contrary to expectation, we detected proteins from all sources in fecal samples of both germ-free and conventional mice suggesting that even protein sources with a high digestive efficiency make it in part to the colon where they can serve as a substrate for the microbiota. Additionally, we found clear patterns where specific component proteins of the dietary protein sources were used as a preferred substrate by the microbiota or were not as accessible to the microbiota. We found that specific proteins with functions that could impact host health and physiology were differentially enriched in germ-free or conventionally raised mice. These findings reveal large differences in the fate of dietary protein from various sources in the gut that could explain some of their differential health impacts.

Efficacy of chemical digestion methods to reveal undamaged microplastics from planktonic samples

Standardisation and validation of methods for microplastics research is essential. A major methodological challenge is the removal of planktonic organisms from marine water samples allowing for the identification of microplastics associated to planktonic communities. To improve the reproducibility and accuracy of digestion methods for the removal of planktonic biomass, we compared and modified existing chemical digestion methods. These digestion methods included an acidic digestion using nitric acid, alkaline digestions with potassium hydroxide (alkaline 1 digestion) and sodium hydroxide from drain cleaner (alkaline 2 digestion), an oxidative digestion using sodium dodecyl sulfate with hydrogen peroxide, and an enzymatic digestion using enzyme drain clean pellets. Chemical digestion of three densities of zooplankton communities (high, medium, and low) in the presence of five commonly found environmental microplastic pollutants (polyamide, polyethylene, polyethylene terephthalate, polypropylene, and polystyrene) were performed for each treatment. The chemical treatments were assessed for (i) their digestion efficiency of zooplankton communities by different biomass densities, and (ii) their impact on microplastic particles through the comparison of both chemical (Raman spectroscopy) and physical (length, width, and visual) changes, between the pre-treatment and post-treatment microplastic particles. The alkaline 1, alkaline 2 and oxidative methods demonstrated significantly better digestion efficiency (p < 0.05) than the modified enzymatic and acidic treatments. The acidic, alkaline 1, and alkaline 2, treatments caused the most damages to the microplastic particles. We suggest future studies to implement the oxidative digestion method with sodium dodecyl sulfate and hydrogen peroxide because of its high digestion efficiency, and low damage to microplastic particles. This method is similar to the wet peroxide oxidation digestion method used throughout the literature but can be implemented at a lower cost.

Enhancing Chymotrypsin Activity and Stability of Capillary Immobilized Enzyme Microreactors Using Zeolitic Imidazolate Frameworks as Encapsulation Materials.

Open-tubular immobilized enzyme microreactors (OT-IMERs) are some of the most widely used enzyme reaction devices due to the advantages of simple preparation and fast sample processing. However, the traditional approaches for OT-IMERs preparation had some defects such as limited enzyme loading amount, susceptibility to complex sample interference, and less stability. Here, we report a strategy for the preparation of highly active and stable OT-IMERs, in which the single-stranded DNA-enzyme composites were immobilized in capillaries and then encapsulated in situ in the capillaries via zeolitic imidazolate frameworks (ZIF-L). The phosphate groups of the DNA adjusted the surface potential of the enzyme to negative values, which could attract cations, such as Zn2+, to promote the formation of ZIF-L for enzyme encapsulation. Using chymotrypsin (ChT) as a model enzyme, the prepared ChT@ZIF-L-IMER has higher activity and better affinity than the free enzyme and ChT-IMER. Moreover, the thermal stability, pH stability, and organic solvent stability of ChT@ZIF-L-IMER were much higher than those of free enzyme and ChT-IMER. Furthermore, the activity of ChT@ZIF-L-IMER was much higher than that of ChT-IMER after ten consecutive reactions. To demonstrate the versatility of this preparation method, we replaced ChT with glucose oxidase (GOx). The stability of GOx@ZIF-L-IMER was also experimentally demonstrated to be superior to that of GOx and GOx-IMER. Finally, ChT@ZIF-L-IMER was used for proteolytic digestion analysis. The results showed that ChT@ZIF-L-IMER had a short digestion time and high digestive efficiency compared with the free enzyme. The present study broadened the synthesis method of OT-IMERs, effectively integrating the advantages of metal-organic frameworks and IMER, and the prepared OT-IMERs significantly improved enzyme stability. All of the results indicated that the IMER prepared by this method had a broad application prospect in capillary electrophoresis-based high-performance enzyme analysis.

Synergistic digestibility effect by planktonic natural food and habitat renders high digestion efficiency in agastric aquatic consumers

A digestibility enhancing effect of natural food on stomachless fish model (Cyprinus carpio) was verified by fluorogenic substrate assays of enzymatic activities in experimental pond carp gut flush and planktonic food over a full vegetative season. Then compared with size-matched conspecific grown artificially (tank carp) and an advanced omnivore species possessing true stomach (tilapia, Oreochromis niloticus). Results suggested activities of digestive enzymes (except amylolytic) were significantly higher in pond carp (p ≤ 0.05) than in the size-matched tank carp. Even compared to tilapia, pond carp appeared superior (p < 0.05; proteolytic or chitinolytic activities) or comparable (p > 0.05; phosphatase or cellulolytic activities). Amylolytic, chitinolytic, and phosphatases activities in pond carp gut significantly increased (p ≤ 0.01) over season. Several orders-of-magnitude higher enzymatic activities were detected in planktonic natural food than expressed in carp gut. Amino acid markers in planktonic food revealed a higher share of zooplankton (microcrustaceans), but not phytoplankton, synchronized with higher activities of complex polysaccharide-splitting enzymes (cellulolytic and chitinolytic) in fish gut. Periods of clear water phase low in chlorophyll-a and nutrients, but high in certain zooplankton (preferably cladocerans), may create a synergistic digestibility effect in pond carp. We conclude aquatic ecosystem components (natural food, water, microbiota) enhance fishes' hydrolyzing capabilities of C/N/P macromolecules and even their complex polymers such as cellulose, chitin, and maybe phytate (to be validated), to the extent that being stomachless is not an issue. Aquatic nutritional ecologists may consider that laboratory-based understandings of digestibility may underestimate digestion efficiency of free-ranging fish in ponds or lakes.

Heat 'n Beat: A Universal High-Throughput End-to-End Proteomics Sample Processing Platform in under an Hour.

Proteomic analysis by mass spectrometry of small (≤2 mg) solid tissue samples from diverse formats requires high throughput and comprehensive proteome coverage. We developed a nearly universal, rapid, and robust protocol for sample preparation, suitable for high-throughput projects that encompass most cell or tissue types. This end-to-end workflow extends from original sample to loading the mass spectrometer and is centered on a one-tube homogenization and digestion method called Heat 'n Beat (HnB). It is applicable to most tissues, regardless of how they were fixed or embedded. Sample preparation was divided into separate challenges. The initial sample washing and final peptide cleanup steps were adapted to three tissue sources: fresh frozen (FF), optimal cutting temperature (OCT) compound embedded (FF-OCT), and formalin-fixed paraffin embedded (FFPE). Third, for core processing, tissue disruption and lysis were decreased to a 7 min heat and homogenization treatment, and reduction, alkylation, and proteolysis were optimized into a single step. The refinements produced near doubled peptide yield when compared to our earlier method ABLE delivered a consistently high digestion efficiency of 85-90%, reported by ProteinPilot, and required only 38 min for core processing in a single tube, with the total processing time being 53-63 min. The robustness of HnB was demonstrated on six organ types, a cell line, and a cancer biopsy. Its suitability for high-throughput applications was demonstrated on a set of 1171 FF-OCT human cancer biopsies, which were processed for end-to-end completion in 92 h, producing highly consistent peptide yield and quality for over 3513 MS runs.

Biochemical and biohythane production from anaerobically digested water plant by hydrothermal liquefaction/gasification

Water hyacinth (Eichhornia crassipes) was anaerobically digested with waste sludge in a batch system at varying total solid (TS) contents (3.3–8.3%) and temperatures (35–55 °C). Then, the high organic content of digested biomass was utilized for hydrothermal liquefaction/gasification in the batch reactor system at different temperatures (200–600 °C) to yield biofuels and biochemicals. Hydrothermal liquefaction/gasification was performed in sub- and super-critical water (above 374 °C and 221 bar) conditions. Prevailing products are some biochemical compounds (carboxylic acids, furfurals, aldehyde/ketones, phenols etc.) at lower temperatures (200–300 °C) while biohythane (total of hydrogen and methane) gaseous fuel are produced at higher temperatures (400–600 °C). The highest carbon gasification efficiency (73 g C in product/g C in feed) was obtained at 600 °C with the sample that has a high anaerobic digestion efficiency (digested at 6.3 TS% at 35 °C). The highest carbon liquefaction efficiency (37.2 g C in product/g C in feed) was obtained at 200 °C with the sample which has a low anaerobic digestion yield (digested at 8.3 TS% at 35 °C).

Investigation of Microplastics (≥10 μm) in Meconium by Fourier Transform Infrared Microspectroscopy.

Microplastics are prevalent emerging pollutants with widespread distribution in air, land and water. They have been detected in human stool, blood, lungs, and placentas. However, human fetal microplastic exposure remains largely under-studied. To assess fetal microplastic exposure, we investigated microplastics using 16 meconium samples. We used hydrogen peroxide (H2O2), nitric acid (HNO3) and a combination of Fenton's reagent and HNO3 pretreatment methods respectively to digest the meconium sample. We analyzed 16 pretreated meconium samples with an ultra-depth three-dimensional microscope and Fourier transform infrared microspectroscopy. The result showed that H2O2, HNO3 and Fenton's reagent combined with HNO3 pretreatment methods could not digest our meconium samples completely. Alternatively, we developed a novel approach with high digestion efficiency using petroleum ether and alcohol (4:1, v/v), HNO3 and H2O2. This pretreatment method had good recovery and non-destructive advantages. We found no microplastics (≥10 μm) in our meconium samples, indicating that microplastic pollution levels in the fetal living environment are miniscule. Different results between previous studies' and ours underscore that comprehensive and strict quality control are necessary for further studies on microplastic exposure using human bio-samples.

In situ digestion of alcohol-fixed cells for quantitative proteomics.

Currently, the bottom-up approach, in which proteins are digested by enzymes such as trypsin prior to mass spectrometry, is the mainstream approach in mass spectrometer-based proteomics. In this approach, the enzymatic digestion process strongly affects the reproducibility of protein identification and quantification. Here, we quantitatively evaluated the enzymatic digestion of proteins under various conditions by quantitative proteomics using data-independent acquisition and found that proteins precipitated with acetone after solubilization with SDS were fully digestible without re-solubilization. This result implies that organic solvent treatment makes cells amenable to trypsin digestion. Direct trypsin digestion of methanol-fixed cells achieved the same digestion efficiency and quantitative reproducibility as the conventional method. Furthermore, this method was found to be equally applicable to mouse liver samples. The establishment of this method indicates that the sample preparation process in bottom-up proteomics can be simplified while maintaining high digestion efficiency and is expected to become a general method for sample preparation in bottom-up proteomics in the future.

Human Hepatic Transporter Signature Peptides for Quantitative Targeted Absolute Proteomics: Selection, Digestion Efficiency, and Peptide Stability.

Quantitative targeted absolute proteomics (QTAP) quantifies proteins by measuring the signature peptides produced from target proteins by trypsin digestion. The selection of signature peptides is critical for reliable peptide quantification. The purpose of this study was to comprehensively assess the digestion efficiency and stability of tryptic peptides and to identify optimal signature peptides for human hepatic transporters and membrane marker proteins. The plasma membrane fraction of the human liver was digested at different time points and the peptides were comprehensively quantified using quantitative proteomics. Transporters and membrane markers were quantified using the signature peptides by QTAP. Tryptic peptides were classified into clusters with low digestion efficiency, low stability, and high digestion efficiency and stability. Using the cluster information, we found that a proline residue next to the digestion site or the peptide position in or close to the transmembrane domains lowers digestion efficiency. A peptide containing cysteine at the N-terminus or arginine-glycine lowers peptide stability. Based on this information and the time course of peptide quantification, optimal signature peptides were identified for human hepatic transporters and membrane markers. The quantification of transporters with multiple signature peptides yielded consistent absolute values with less than 30% of coefficient variants in human liver microsomes and homogenates. The signature peptides selected in the present study enabled the reliable quantification of human hepatic transporters. The QTAP protocol using these optimal signature peptides provides quantitative data on hepatic transporters usable for integrated pharmacokinetic studies.

Extraction of biodegradable microplastics from tissues of aquatic organisms

Biodegradable plastics (BPs) have been given high hopes to substitute conventional plastics, but their biodegradation requires strict conditions. BPs can accumulate for a long time in the environment and even derive biodegradable microplastics (BMPs), thus threatening wildlife and ecosystems. However, no efficient method is available for extracting BMPs from organisms' tissues. This study used multi-criteria decision-making (MCDM) methods to comprehensively evaluate and optimize extraction protocols of five BMPs from economic aquatic species. Digestion time, digestion efficiency, mass loss, cost, polymer integrity and size change were selected as evaluating indictors. According to the screening results of MCDM methods, Pepsin+H2O2 was selected as the optimal digestion method of BMPs because of its highest comprehensive score, which has high digestion efficiency (99.56%) and minimum plastic damage. Compared with olive oil, NaI is more suitable for separating BMPs from the digested residues. Furthermore, the combination of Pepsin+H2O2 digestion and NaI density separation was used to extract all five kinds of BMPs from the bivalve, crab, squid, and crayfish tissues, and all the recovery rates exceeded 80%. These results suggest that the optimal protocol is practicable to extract various BMPs from various aquatic organisms.

Effect of salinity on the determination of dissolved non-reactive phosphorus and total dissolved phosphorus in coastal waters.

The salinity may affect the phosphorus (P) determination accuracy in coastal waters, especially for the dissolved non-reactive P (DNRP) and total dissolved P (TDP). In this work, the competition mechanism between NaCl and DNRP for oxidants (K2 S2 O8 , the most commonly used and recognized oxidant) was identified in different DNRP determinations. Furthermore, salinity influences on determinations of tetrasodium pyrophosphate decahydrate, glyphosate, phytic acid sodium salt hydrate, adenosine-5'-nomophosphate disodium, and TDP were investigated. The results indicated that approximately 10% IHP6 and AMP would be transferred to dissolved reactive P (DRP) during digestion without K2 S2 O8 . When NaCl increased from 0% to 3.5% with fresh water method, the determination of Gly + K2 HPO4 and IHP6 + K2 HPO4 decreased by 8.0% ± 0.00% and 24% ± 0.01%, respectively. In addition, the determinations of DNRPs and TDP with different salinities in natural coastal waters by fresh water method and seawater method were performed. It showed that when the salinity >5.0 PSU, the DNRPs and TDP determination results presented deviations. At a salinity of 35.0 PSU, the TDP (KH2 PO4 + Gly + IHP6 + AMP) reduction measured by two methods was more than 12.3% ± 0.46%. Furthermore, oxidants with higher digestion efficiency than K2 S2 O8 should be developed. PRACTITIONER POINTS: ~10% IHP6 and AMP could be transferred to DRP during digestion without K2 S2 O8 addition. Salinity affects the DNRPs determination results mainly due to competition for oxidants and complexation with metal ions. More than 12.3% TDP in coastal waters could not be measured when the salinity was 35.0 PSU.

Development of Optimal Digesting Conditions for Microplastic Analysis in Dried Seaweed Gracilaria fisheri.

Currently, research on the accumulation of microplastics (MPs) in the marine food web is being highlighted. An accurate and reliable digestion method to extract and isolate MPs from complex food matrices has seldom been validated. This study aimed to compare the efficacy of MP isolation among enzymatic-, oxidative-, and the combination of two digestion methods on red seaweed, Gracilaria fisheri. The dried seaweed sample was digested using three different methods under various conditions using enzymes (cellulase and protease), 30% H2O2, and a combination of enzymes and 30% H2O2. The method possessing the best digestion efficiency and polymer recovery rate of MPs was selected, and its effect on spiked plastic polymer integrity was analyzed by Raman spectroscopy. As a result, the enzymatic method rendered moderate digestion efficiency (59.3–63.7%) and high polymer recovery rate (94.7–98.9%). The oxidative method using 30% H2O2 showed high digestion efficiency (93.0–96.3%) and high polymer recovery rate (>98%). The combination method was the most effective method in terms of digestion efficiency, polymer recovery rate, and expenditure of digestion time. The method also showed no chemical changes in the spiked plastic polymers (PE, PP, PS, PVC, and PET) after the digestion process. All the spiked plastic polymers were identifiable using Raman spectroscopy.

Nano-selenium on reproduction and immunocompetence: an emerging progress and prospect in the productivity of poultry research.

Nanotechnology, an emerging and promising technology, has been implicated to revolutionize the poultry industry. The main aspect of nanotechnology was to modify or alter the particle size into nanometers and thereby alter the physical as well as chemical features of the particular molecules. Selenium (Se), an essential trace element, can play an immense role in the maintenance of diverse physiological functions, body metabolism and cellular homeostasis, and the performance of poultry. Selenium nanoparticles (Se-NPs) are of growing importance due to its nutrients digestibility, medicinal therapy, targeted drug delivery system, and production of vaccines. Se-nanoparticles are having importance due to its high bioavailability and digestive efficiency. Se-NPs have been implicated to increase relative weights of immune-related organs (burse and thymus) to enhance immunity and thereby modulate egg production as well as the reproductive performance of birds. The present review is highlighted on the significant role of nano-selenium on reproductive performance and immunocompetence in poultry as comparative advantages over conventional sources of Se in poultry diets.

The effects of acute temperature change and digestive status on in situ cardiac function in mahi-mahi (Coryphaena hippurus)

In this study, we investigated the effect of acute increases in temperature on cardiovascular function of mahi-mahi (Coryphaena hippurus). We also describe, for the first time, an artery that supplies the gastrointestinal tract that originates from the fourth branchial artery. We used vascular casting to verify the anatomical location of this unique celiaco-mesenteric artery. We predicted that blood flow in this vessel would be correlated with the digestive state of the animal. Increasing water temperature from 25.0 to 30.5 °C resulted in a linear increase in heart rate (fH) from 165 ± 4 beats∙min−1to 232 ± 7 beats∙min−1. Over this temperature range, fH strongly correlated with water temperature (R2 = 0.79). At 31 °C fH no longer correlated with water temperature, and at 34 °C fH had dropped to 114 ± 19 beats∙min−1. Furthermore, we found that mahi are capable of maintaining constant cardiac output over a temperature range from 25 to 31 °C. Cardiac function appeared to be compromised at temperatures >31 °C. In fed anesthetized fish, blood flow was pulsatile in the celiaco-mesenteric artery and was not in fasted fish. In fed fish, blood flow in the left celiaco-mesenteric artery was 1.99 ± 0.78 ml·min−1·kg−1 compared to the total cardiac output of 168.6 ± 12.7 ml·min−1·kg−1. The data suggest that mahi can differentially regulate gastric blood flow based on feeding state, which may explain the high digestive efficiency and very high growth rates of these pelagic predators.

Effects of Orange Pulp Conservation Methods (Dehydrated or Ensiled Sun-Dried) on the Nutritional Value for Finishing Pigs and Implications on Potential Gaseous Emissions from Slurry.

Simple SummaryUtilization of local by-products in pig nutrition can reduce the environmental impact of feeds and contribute to the sustainable development of the livestock sector. Orange pulp (OP) is the most abundant citrus by-product worldwide, but its seasonal production and perishable nature requires storage and drying procedures that might affect its nutritive value. Conservation process by fuel drying is expensive and can impair feed sustainability. Instead, in the Mediterranean countries, OP is sun-dried in the open-air. This procedure often implies a previous silage (during storage) which occurs naturally, because OP has a high level of sugars available for fermentation. Orange pulp is also rich in soluble fiber, which is highly fermentable at the pig’s caecum and may reduce gas emissions from slurry. In this study, the nutritive value of conventional fuel-dehydrated (DOP) or ensiled-sun dried (ESDOP) was determined for pig diets. Sugars fermentation during ensiling increases fiber level in ESDOP and decreases energy digestibility compared to DOP, but both OP have an appreciable digestible energy content for pigs, around 87 and 94% that of barley, respectively. In addition, they do not differ in the amount of slurry excreted and contribute to reduce potential derived ammonia and methane emissions.The inclusion of orange pulp (OP) in pig diets may promote the circular economy, but drying procedures might influence its nutritional value and environmental impact. The purpose of this study was to determine the energy value and nutrient digestibility of dehydrated (DOP) and ensiled sun dried (ESDOP) orange pulp. The potential ammonia (NH3) and methane (CH4) emissions derived from slurry were also measured. Digestible energies of 14.2 and 13.2 MJ/kg DM for DOP and ESDOP, respectively, were estimated by difference after a 500 g/kg substitution of a basal diet with OPs. A high fiber digestion efficiency was observed for both OPs. Pigs fed the basal diet showed a higher intake and a greater excretion of urine N than pigs fed with OP, but fecal N excretion did not differ among diets. A higher benzoic and hippuric acid content in urine was observed in OP than in basal diet. Altogether, these findings explained a lower pH in slurry of OP diets and a reduction of potential NH3 emissions. The biochemical CH4 potential also decreased, especially with ESDOP. Overall, both OP are relevant sources of energy for pig diets. Their inclusion in feeds generate favorable changes of slurry characteristics that reduce potential NH3 and CH4 emissions.

Microwave-based strategies for sample preparation and halogen determination in blood using ICP-MS

In this work, two microwave-assisted sample preparation methods based on combustion and ultraviolet digestion for futher determination of Br, Cl, and I in blood by inductively coupled plasma mass spectrometry were proposed. For microwave-induced combustion (MIC), blood was spotted on square pieces of Munktell TFN paper, and water and diluted NH4OH were evaluated as absorbing solutions. The stability of the analytes in the spotted blood was also evaluated. During 90 days, no changes of halogen content were observed for samples (with and without anticoagulant) on paper stored in a desiccator at 20 ± 5 °C. Whole blood was also digested by microwave-assisted ultraviolet digestion (MAD-UV). Some parameters as the volume of HNO3 solution, the influence of H2O2, and the effect of UV radiation were investigated. The interferences caused by C and K on halogen determination were carefully investigated. The results for MIC and MAD-UV and Br, Cl and I determination by ICP-MS presented no difference. The accuracy of MIC and MAD-UV was also evaluated by analyte spike with reference solutions of Br, Cl and I using inorganic species and also organic I standard (T4-levothyroxine). For both methods, recoveries were 94 and 108% for Br, 96 and 103% for Cl, and 104 and 97% for I. In case of organic I, recoveries were 99 and 111% for MIC and MAD-UV, respectively. The limits of quantification for MIC and MAD-UV after ICP-MS determination were 0.06 and 0.04 μg g−1 for Br, 14 and 30 μg g−1 for Cl and 12 and 8 ng g−1 for I, respectively. The proposed methods provide a suitable digestion approach, assuring safety and high digestion efficiency for further halogen determination in blood, with the possibility to use in clinical analysis.

Enzymatic purification of microplastics in soil

Accurate and effective analysis of microplastics depends on the purification of any biogenic matter present in the environmental sample. Within the soil environment, the presence of biogenic materials (living and non-living) obstruct reliable identification of microplastics. However, while conventional acidic, alkaline, and wet peroxidation methods are often used for microplastics purification, they can result in significant alteration to the polymer integrity. To overcome this issue of polymer damage, we have developed a more efficient protocol using enzymes to eliminate organic materials from soil media without affecting the synthetic polymers. This method describes a simple enzymatic protocol that eliminates the organic matter based on the application of enzymes that target the three natural components of lignocellulosic biomass (cellulose, hemicellulose, and lignin). The enzymatic protocol introduced in this study allows for the use of an oxidizing agent for the pre-treatment of samples and density separation step. Additionally, this method was used to determine the enzymatic digestion efficiencies of soil organic matter and recovery rates of microplastics. Finally, the recovered microplastics were characterized by techniques including stereomicroscopy, FT-IR, and SEM.This method proved to be effective in reducing approximately >90% of organic materials in soil samples. After showing a high digestion efficiency the method was applied to spiked microplastic soil samples where high recovery rates were established and plastic characteristics were conserved. Despite being a time consuming and expensive method when compared to other purification methods. The key benefits of this methodology are:•Minimal sample preparation•Representative purification of microplastic from diverse soil media; and•Most importantly, preservation of polymer integrity aiding requirements needed for particle identification (e.g.,% mass polymer remaining, SEM images revealing no obvious polymer morphology change after enzyme treatment).

Phenotypic timeline of gastrointestinal tract development in broilers divergently selected for digestive efficiency

Sustainability of poultry farming relies on the development of more efficient and autonomous production systems in terms of feed supply. This implies a better integration of adaptive traits in breeding programs, including digestive efficiency, to favor the use of a wider variety of feedstuffs. The objective of the study was to better characterize the kinetics of development of the digestive tract in broilers, in relationship with digestive efficiency by measuring various digestive parameters as well as serum color. Absolute and relative growth of gastrointestinal tract organs were compared between 2 divergent chicken lines selected on digestive efficiency (AMEn) during 7 wk of development. We show that as early as 7 d of age, these 2 lines differs for several organs developments and that these differences remain visible later on. In addition, the allometry of the gizzard and intestine segments is different between the 2 lines, with efficient birds putting more effort in the upper part of the digestive tract during postnatal development and less-efficient birds putting more effort in the lower part of the gastrointestinal tract. Interestingly, we also showed that differences in serum pigmentation, which is a good biomarker for digestive capacity, could be a convenient diagnostic tool to discriminate between chickens with high or low digestive efficiency at early stages of development. In conclusion, this study showed that selection of chickens for AMEn had large impacts in gastrointestinal development including at early stages and is a valuable resource for further studies on the genetic and physiological control of the response of the animal to feed variations.

Peculiar Paramecium Hosts Fail to Establish a Stable Intracellular Relationship With Legionella pneumophila.

Legionella pneumophila, an intracellular human pathogen, establishes intracellular relationships with several protist hosts, including Paramecium caudatum. L. pneumophila can escape the normal digestion process and establish intracellular relationships in Paramecium. In this study, we identify new Paramecium strains that significantly reduce the number of L. pneumophila during infection. As a result, stable intracellular relationships between L. pneumophila and these Paramecium strains were not observed. These digestion-type Paramecium also showed high efficiency for Escherichia coli elimination compared to other strains of Paramecium. These results suggest that the digestion-type strains identified have high non-specific digestion activity. Although we evaluated the maturation process of Legionella-containing vacuoles (LCVs) in the Paramecium strains using LysoTracker, there were no discriminative changes in these LCVs compared to other Paramecium strains. Detailed understanding of the mechanisms of high digestion efficiency in these strains could be applied to water purification technologies and L. pneumophila elimination from environmental water.