Digesting the Indigestible: Microplastic Extraction from Prawn Digestive Tracts

Microplastics (MPs) have become ubiquitous in the marine environment, and are likely ingested by a broad cross-section of marine life. The extent to which marine organisms ingest MPs is uncertain due to limitations in analytical methods. Effective identification and analysis of ingested MPs is a precursor to understand their impact on marine organisms and their human consumers. This is particularly challenging for crustaceans, due to the chitin present in their exoskeleton and digestive systems, which is resistant to chemical degradation. This study presents a novel application that can efficiently break down the stable organic tissue of banana prawns (Penaeus merguiensis), and subsequently isolate putative MP polymers from the digestive tract without damaging their integrity. Five treatments were examined for their capacity to break down chitin from the prawn digestive system; namely acid, alkaline, oxidant, enzyme and microwave assisted oxidant digestion. Gravimetric and image analysis revealed that the organic tissue of the prawn gastrointestinal tract can be effectively removed by acid, oxidant, and microwave assisted oxidant digestion methods. However, testing on seven reference polymers (polyamide (PA), polyethylene (PE), polyester (PES), polypropylene (PP), polystyrene (PS), polyvinyl chloride (PVC), and rayon) revealed significant degradation when exposed to acid digestion. Overall, microwave assisted oxidant digestion achieved the best recovery rate of spiked MPs ( > 90%) with minimal size, shape, and Fourier transform infrared (FTIR) spectral changes for all polymers except for rayon. These results highlight a new direction for tissue removal and MP extraction in crustacean ingestion studies

Plastics for dinner: Store-bought seafood, but not wild-caught from the Great Barrier Reef, as a source of microplastics to human consumers

Seafood accounts for more than 17% of the global consumption of animal protein, with an excess of 335000 t consumed in Australia throughout 2019-2020. Recently, the presence of microplastics (MPs) within commercial seafood and the potential vectorisation of MPs to human consumers has become a significant concern for the public and the scientific community. Here, four commonly harvested wild-caught marine organisms were assessed for MP presence. These species comprise a significant proportion of the Queensland seafood industry, as well as being highly desirable to Australian consumers. The edible muscle tissue and discarded digestive tissue (GIT) of barramundi (Lates calcifer), coral trout (Plectropomus leopardus), blue leg king prawns (Melicertus latisulcatus), and Ballot's saucer scallops (Ylistrum balloti), were analysed discretely to determine the extent to which these species may be contaminated in the wild (GIT tissue), and the extent to which they themselves may act as a vector for human exposure (edible muscle tissue). Wild-caught seafood was predominantly free of MPs, with digestive tissues from two of ten coral trout containing only two fibres each. All wild-caught muscle tissue samples were free of MPs, as was the GIT of scallops, prawns, and barramundi. On the other hand, fresh, skinless barramundi muscle tissues, purchased from various commercial suppliers, were examined and found to be significantly contaminated with MPs (0.02 - 0.19 MP g-1). Overall, these results highlight the growing consensus that food can become contaminated simply by being prepared in the human environment, and the focus must shift to determining the extent of MP proliferation within the processing and point-of-sale environment

Taking control of microplastics data: A comparison of control and blank data correction methods

Although significant headway has been achieved regarding method harmonisation for the analysis of microplastics, analysis and interpretation of control data has largely been overlooked. There is currently no consensus on the best method to utilise data generated from controls, and consequently many methods are arbitrarily employed. This study identified 6 commonly implemented strategies: a) No correction; b) Subtraction; c) Mean Subtraction; d) Spectral Similarity; e) Limits of detection/ limits of quantification (LOD/LOQ) or f) Statistical analysis, of which many variations are possible. Here, the 6 core methods and 45 variant methods (n = 51) thereof were used to correct a dummy dataset using control data. Most of the methods tested were too inflexible to account for the inherent variation present in microplastic data. Only 7 of the 51 methods tested (six LOD/LOQ methods and one statistical method) showed promise, removing between 96.3 % and 100 % of the contamination data from the dummy set. The remaining 44 methods resulted in deficient corrections for background contamination due to the heterogeneity of microplastics. These methods should be avoided in the future to avoid skewed results, especially in low abundance samples. Overall, LOD/LOQ methods or statistical analysis comparing means are recommended for future use in microplastic studies

Safe food through better labelling: A robust method for the rapid determination of caprine and bovine milk allergens

Accidental milk cross-contamination is one of the most common causes for costly food recalls. Yet, quantifying trace-levels of allergen is time-consuming and current methods are not adapted for routine analyses making quality control for trace-level allergen content impractical. This perpetuates voluntary “may-contain” statements that are unhelpful for people suffering from food allergies. Here, we developed a rapid LC-MS method enabling milk allergen quantification by comparing all tryptic-peptides of major milk allergens. The bovine-specific αS-2 casein peptide and allergen-epitope NAVPITPTLNR provided excellent performance in sensitivity (LOD 1 mg.kg − 1; LOQ 2 mg.kg − 1) across various dairy products, good recovery rates in baked croissants (77 % with a 10 % inter-day RSD) and a linear range of 2 – 2,000 mg.kg − 1. The method can be used for routine determination of trace-contamination with bovine milk allergen and the adulteration of high-value caprine dairy products with lower-value bovine milk products, protecting consumer trust and the growing population suffering from food allergies

Ingestion and depuration of microplastics by a planktivorous coral reef fish, Pomacentrus amboinensis

Microplastics are ubiquitous contaminants in marine environments and organisms. Concerns about potential impacts on marine organisms are usually associated with uptake of microplastics, especially via ingestion. This study used environmentally relevant exposure conditions to investigate microplastic ingestion and depuration kinetics of the planktivorous damselfish, Pomacentrus amboinensis. Irregular shaped blue polypropylene (PP) particles (longest length 125–250 μm), and regular shaped blue polyester (PET) fibers (length 600–700 μm) were selected based on physical and chemical characteristics of microplastics commonly reported in the marine environment, including in coral reef ecosystems. Individual adult damselfish were exposed to a single dose of PP particles and PET fibers at concentrations reported for waters of the Great Barrier Reef (i.e., environmentally relevant concentrations, ERC), or future projected higher concentrations (10x ERC, 100x ERC). Measured microplastic concentrations were similar to their nominal values, confirming that PP particles and PET fibers were present at the desired concentrations and available for ingestion by individual damselfish. Throughout the 128-h depuration period, the 88 experimental fish were sampled 2, 4, 8, 16, 32, 64, and 128-h post microplastic exposure and their gastrointestinal tracts (GIT) analyzed for ingested microplastics. While damselfish ingested both experimental microplastics at all concentrations, body burden, and depuration rates of PET fibers were significantly larger and longer, respectively, compared to PP particles. For both microplastic types, exposure to higher concentrations led to an increase in body burden and lower depuration rates. These findings confirm ingestion of PP particles and PET fibers by P. amboinensis and demonstrate for the first time the influence of microplastic characteristics and concentrations on body burden and depuration rates. Finally, despite measures put in place to prevent contamination, extraneous microplastics were recovered from experimental fish, highlighting the challenge to completely eliminate contamination in microplastic exposure studies. These results are critical to inform and continuously improve protocols for future microplastics research, and to elucidate patterns of microplastic contamination and associated risks in marine organisms

Application of near-infrared spectroscopy and chemometrics for the rapid detection of insect protein adulteration from a simulated matrix

The popularity of insect protein as a food and feed supplement is growing. Protein quality, end use and prices vary considerably between different insect species, which may incentivise insect protein adulteration. Here, near-infrared (NIR) spectroscopy and chemometrics were used to detect the presence of cricket, black soldier fly larvae (BSFL) and mealworm proteins in a simulated complex insect protein mixture. Additionally, BSFL protein powders collected from three commercial sources were investigated to determine whether the NIR-based technology can discriminate the proteins obtained from different companies based on their composition. The proximate analysis suggests compositional protein, fat and chitin differences between insect species. A partial least square (PLS) regression model obtained Q2 values ranging from 0.991 to 0.997 for the predictions of the content of protein mixtures containing BSFL, cricket and mealworm powders mixed at various proportions. The root mean square error of cross-validation (RMSCV) values range from 1.8% to 2.9%, and residual prediction deviation (RPD) values from 10.4 to 17.1 for the adulterated insect protein powders. The accuracy of the prediction model (∼2%) for the adulterated percentages varied depending on the insect species. The NIR spectra could differentiate (Q2 = 0.999) the origin of BSFL protein powders from three different companies and two types of processing (whole meal and defatted samples). Overall, this study established a rapid and low-cost insect protein adulteration monitoring pipeline for the three common insect protein powders. We envisage that NIR can be applied to assess insect adulteration, authentication, and quality control in the emerging insect food and feed industries

Common activation of canonical Wnt signaling in pancreatic adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDA) is an extremely aggressive malignancy, which carries a dismal prognosis. Activating mutations of the Kras gene are common to the vast majority of human PDA. In addition, recent studies have demonstrated that embryonic signaling pathway such as Hedgehog and Notch are inappropriately upregulated in this disease. The role of another embryonic signaling pathway, namely the canonical Wnt cascade, is still controversial. Here, we use gene array analysis as a platform to demonstrate general activation of the canonical arm of the Wnt pathway in human PDA. Furthermore, we provide evidence for Wnt activation in mouse models of pancreatic cancer. Our results also indicate that Wnt signaling might be activated downstream of Hedgehog signaling, which is an early event in PDA evolution. Wnt inhibition blocked proliferation and induced apoptosis of cultured adenocarcinoma cells, thereby providing evidence to support the development of novel therapeutical strategies for Wnt inhibition in pancreatic adenocarcinoma

Marine Mammal Brucella Genotype Associated with Zoonotic Infection

Marine Mammal Brucella Genotype Associated with Zoonotic Infectio

Mapping and identification of candidate loci responsible for Peromyscus hybrid overgrowth

Crosses between two recently diverged rodent species of the genus Peromyscus result in dramatic parent-of-origin effects on growth and development. P. maniculatus females crossed with P. polionotus males yield growth-retarded conceptuses, whereas the reciprocal cross results in overgrowth and lethality. These hybrid effects are particularly pronounced in the placenta. We previously detected linkage to two regions of the genome involved in the overgrowth effects. One locus, termed Peal, is a paternally expressed autosomal locus mapping to a domain whose house mouse equivalent contains several clusters of imprinted genes. The other locus, termed Mexl, maps to a gene-poor region of the X chromosome. Here we use an advanced intercross line to verify and narrow the regions of linkage and identify candidate genes for Mexl and Peal. While we have previously shown that Mexl affects both pre-and postnatal growth, we show here that Peal affects only prenatal growth. Utilizing criteria such as mutant phenotypes and allelic expression, we identify the loci encoding the homeobox protein Esx1 and the zinc-finger protein Pw1/Peg3 as candidates. Both loci exhibit expression changes in the hybrids

Use of the oral beta blocker bisoprolol to reduce the rate of exacerbation in people with chronic obstructive pulmonary disease (COPD): a randomised controlled trial (BICS)

Background: Chronic obstructive pulmonary disease (COPD) is associated with significant morbidity, mortality and healthcare costs. Beta blockers are well-established drugs widely used to treat cardiovascular conditions. Observational studies consistently report that beta blocker use in people with COPD is associated with a reduced risk of COPD exacerbations. The bisoprolol in COPD study (BICS) investigates whether adding bisoprolol to routine COPD treatment has clinical and cost-effective benefits. A sub-study will risk stratify participants for heart failure to investigate whether any beneficial effect of bisoprolol is restricted to those with unrecognised heart disease. Methods: BICS is a pragmatic randomised parallel group double-blind placebo-controlled trial conducted in UK primary and secondary care sites. The major inclusion criteria are an established predominant respiratory diagnosis of COPD (post-bronchodilator FEV1 < 80% predicted, FEV1/FVC < 0.7), a self-reported history of ≥ 2 exacerbations requiring treatment with antibiotics and/or oral corticosteroids in a 12-month period since March 2019, age ≥ 40 years and a smoking history ≥ 10 pack years. A computerised randomisation system will allocate 1574 participants with equal probability to intervention or control groups, stratified by centre and recruitment in primary/secondary care. The intervention is bisoprolol (1.25 mg tablets) or identical placebo. The dose of bisoprolol/placebo is titrated up to a maximum of 4 tablets a day (5 mg bisoprolol) over 4–7 weeks depending on tolerance to up-dosing of bisoprolol/placebo—these titration assessments are completed by telephone or video call. Participants complete the remainder of the 52-week treatment period on the final titrated dose (1, 2, 3, 4 tablets) and during that time are followed up at 26 and 52 weeks by telephone or video call. The primary outcome is the total number of participant reported COPD exacerbations requiring oral corticosteroids and/or antibiotics during the 52-week treatment period. A sub-study will risk stratify participants for heart failure by echocardiography and measurement of blood biomarkers. Discussion: The demonstration that bisoprolol reduces the incidence of exacerbations would be relevant not only to patients and clinicians but also to healthcare providers, in the UK and globally. Trial registration: Current controlled trials ISRCTN10497306. Registered on 16 August 201