Freshwater microalgae Nannochloropsis limnetica for the production of ß-galactosidase from whey powder.

This study investigated the first-ever reported use of freshwater Nannochloropsis for the bioremediation of dairy processing side streams and co-generation of valuable products, such as ß-galactosidase enzyme. In this study, N. limnetica was found to grow rapidly on both autoclaved and non-autoclaved whey-powder media (referred to dairy processing by-product or DPBP) without the need of salinity adjustment or nutrient additions, achieving a biomass concentration of 1.05-1.36 g L-1 after 8 days. The species secreted extracellular ß-galactosidase (up to 40.84 ± 0.23 U L-1) in order to hydrolyse lactose in DPBP media into monosaccharides prior to absorption into biomass, demonstrating a mixotrophic pathway for lactose assimilation. The species was highly effective as a bioremediation agent, being able to remove > 80% of total nitrogen and phosphate in the DPBP medium within two days across all cultures. Population analysis using flow cytometry and multi-channel/multi-staining methods revealed that the culture grown on non-autoclaved medium contained a high initial bacterial load, comprising both contaminating bacteria in the medium and phycosphere bacteria associated with the microalgae. In both autoclaved and non-autoclaved DPBP media, Nannochloropsis cells were able to establish a stable microalgae-bacteria interaction, suppressing bacterial takeover and emerging as dominant population (53-80% of total cells) in the cultures. The extent of microalgal dominance, however, was less prominent in the non-autoclaved media. High initial bacterial loads in these cultures had mixed effects on microalgal performance, promoting ß-galactosidase synthesis on the one hand while competing for nutrients and retarding microalgal growth on the other. These results alluded to the need of effective pre-treatment step to manage bacterial population in microalgal cultures on DPBP. Overall, N. limnetica cultures displayed competitive ß-galactosidase productivity and propensity for efficient nutrient removal on DPBP medium, demonstrating their promising nature for use in the valorisation of dairy side streams.

Pectin/sodium alginate-based active film integrated with microcrystalline cellulose and geraniol for food packaging applications.

Biopolymer-based packaging films were prepared from pectin (PEC) and sodium alginate (SA), with the incorporation of 10 % MCC and different concentrations of geraniol (GER at 2.5, 5.0, 7.5, and 10.0 %). Rheological properties suggested that film-forming solutions and film-forming emulsions exhibited a shear-thinning or pseudo-plastic non-Newtonian behaviour. The dried films were crosslinked with 2.0 % CaCl2. The addition of MCC into PEC/SA film enhanced the TS but reduced it with the impregnation of GER without influencing the EAB and toughness of the film. The water solubility of the films significantly reduced with the rise in the GER levels but enhanced the water vapor and oxygen barrier attributes. TGA demonstrated that incorporating MCC reduced the film's thermal degradation (44.92 % to 28.81 %), but GER had an insignificant influence on the thermal stability. FTIR spectra revealed that hydrogen bond formation was positively linked with the GER addition in the film formulation. X-ray diffractograms showed that prepared films were predominantly amorphous. Antimicrobial studies showed a complete reduction of Escherichia coli and Bacillus cereus in 24 h. Overall, the composite film displayed excellent physical and active properties and PEC/SA/MCC/5.0 %GER/CaCl2 film was considered the best formulation for food packaging applications.

Assessing the Effects of Cold Atmospheric Plasma on the Natural Microbiota and Quality of Pork during Storage.

Cold atmospheric plasma (CAP) is a novel non-thermal technology with significant potential for use in meat processing to prolong shelf life. The objective of the study was to evaluate the efficiency of CAP treatment on the natural microbiota and quality traits of pork stored for 8 days at 4 °C. CAP treatment was applied by employing piezoelectric direct discharge technology to treat pork samples for 0, 3, 6, and 9 min. Reductions of approximately 0.8-1.7 log CFU/g were observed in total viable counts (TVC) and Pseudomonas spp. levels for CAP treatments longer than 3 min, immediately after treatment. A storage study revealed that CAP-treated pork (>6 min) had significantly lower levels of TVC, Pseudomonas spp., and Enterobacteriaceae throughout storage. Regarding quality traits, CAP application for longer than 3 min significantly increased water retention and yellowness and decreased meat redness compared to untreated pork. However, other parameters such as pH, tenderness, and lightness exhibited no statistically significant differences between untreated and CAP-treated pork. Lipid oxidation levels were higher only for the 9-min treatment compared to untreated pork. Our results revealed that CAP is a promising technology that can extend the microbiological shelf life of pork during refrigeration storage.

Clinical Pattern of Pediatric Hydatid Disease.

Context: Literature regarding hydatid disease in children is sparse. Aims: To highlight the peculiarities in the clinical pattern of pediatric hydatid disease (PHD). Settings and Design/Materials and Methods: Data were collected retrospectively from all children aged <18 years who presented to our tertiary care institute from July 2021 to June 2023 with hydatid disease involving any organ. Statistical Analysis Used: Simple statistical analysis involving sums, means, averages, and percentages. Results: Four of the 10 cases (40%) involved the lung, while only 2 (20%) involved the liver. There were five females and four males with an age range of 2-17 years. Four of the cases had primary extrahepatic extrapulmonary hydatid disease (40%), two involving the pancreas, one in the rectouterine pouch, and one intracranial. Conclusions: The clinical pattern of PHD is different from that of adults. Pulmonary echinococcosis is more common than hepatic involvement. Primary extrahepatic extrapulmonary hydatid disease is also more common in children than previously thought. A cystic lesion anywhere in a child warrants a differential of hydatid disease.

Synthesis of cationic liposome nanoparticles using a thin film dispersed hydration and extrusion method.

Liposome nanoparticles can carry a wide range of therapeutic molecules including small molecules and nucleic acid-based therapeutics. Potential benefits include translocation across physiological barriers, reduced systemic toxicity, and enhanced pharmacokinetic parameters such as absorption, distribution, selective release and optimal elimination kinetics. Liposome nanoparticles can be generated with a wide range of natural and synthetic lipid-based molecules that confer desirable properties depending on the desired therapeutic application Nel et al (2023), Large (2021), Elkhoury (2020). This protocol article seeks to detail the procedures involved in the production of cationic liposomes using thin-film dispersed hydration method with an estimated uniform size of 60-70 nm for targeted drug administration in tumor cells, by modifying the previous one also published by the same authors cited here. The method was carrying out using N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl (DOTAP, 2 mg) as cationic lipid and cholesterol (0.5 mg) in a molar ratio of 7:3 respectively. The liposomal suspension was obtained and its physical, chemical and biological properties were determined. A two-step extrusion process, using 100 nm and 50 nm polycarbonate membranes, was carried. The results demonstrate generation of liposome nanoparticles with a size of 60-70 nm stable for at least 16 weeks and with an encapsulation efficiency of approximately 81% using Doxorubicin.

Upcycling of seafood side streams for circularity.

The upcycling of seafood side streams emerges as a crucial facet in the quest for circularity within the food industry, surpassing other food sources in its significance. Seafood side stream plays an indispensable role in global food security and human nutrition. Nevertheless, losses ensue throughout the seafood supply chain, resulting in substantial waste generation. These underutilized seafood by-products contain valuable resources like edible proteins and nitrogenous compounds. Projections indicate that fishery products' utilization for human consumption will soar to 204 MT by 2030. Yet, the industry annually generates millions of tonnes of waste, predominantly from crab, shrimp, and lobster shells, leading to environmental impacts due to COD and BOD issues. A five-tier circular economic model offers a framework to manage seafood side-streams efficiently, with applications spanning pharmaceuticals, food production, animal feed, fertilizers, and energy fuel, thereby maximizing their potential and reducing waste in line with sustainability goals.

Acoustic cavitation for agri-food applications: Mechanism of action, design of new systems, challenges and strategies for scale-up.

Acoustic cavitation, an intriguing phenomenon resulting from the interaction of sound waves with a liquid medium, has emerged as a promising avenue in agri-food processing, offering opportunities to enhance established processes improving primary production of ingredients and further food processing. This comprehensive review provides an in-depth analysis of the mechanisms, design considerations, challenges and scale-up strategies associated with acoustic cavitation for agri-food applications. The paper starts by elucidating the fundamental principles of acoustic cavitation and its measurement, delving then into the diverse effects of different parameters associated with, the acoustic wave, mechanical design and operation of the ultrasonic system, along with those related to the food matrix. The technological advancements achieved in the design and set-up of ultrasonic reactors addressing limitations during scale up are also discussed. The design, engineering and mathematical modelling of ultrasonic equipment tailored for agri-food applications are explored, along with strategies to maximize cavitation intensity and efficiency in the application of brining, freezing, drying, emulsification, filtration and extraction. Advanced US equipment, such as multi-transducers (tubular resonator, FLOW:WAVE®) and larger processing surface areas through innovative designing (Barbell horn, CascatrodesTM), are one of the most promising strategies to ensure consistency of US operations at industrial scale. This review paper aims to provide valuable insights into harnessing acoustic cavitation's potential for up-scaling applications in food processing via critical examination of current research and advancements, while identifying future directions and opportunities for further research and innovation.

Conventional extraction of fucoidan from Irish brown seaweed Fucus vesiculosus followed by ultrasound-assisted depolymerization.

Fucoidan has attracted considerable attention from scientists and pharmaceutical companies due to its antioxidant, anticoagulant, anti-inflammatory, anti-tumor, and health-enhancing properties. However, the extraction of fucoidan from seaweeds often involves the use of harsh chemicals, which necessitates the search for alternative solvents. Additionally, the high viscosity and low cell permeability of high molecular weight (Mw) fucoidan can limit its effectiveness in drug action, while lower Mw fractions exhibit increased biological activity and are also utilized as dietary supplements. The study aimed to (1) extract fucoidan from the seaweed Fucus vesiculosus (FV) using an environmentally friendly solvent and compare it with the most commonly used extraction solvent, hydrochloric acid, and (2) assess the impact of ultrasound-assisted depolymerization on reducing the molecular weight of the fucoidan extracts and examine the cytotoxic effect of different molecular weight fractions. The findings indicated that the green depolymerization solvent, in conjunction with a brief ultrasound treatment, effectively reduced the molecular weight. Moreover, a significant decrease in cell viability was observed in selected samples, indicating potential anticancer properties. As a result, ultrasound was determined to be an effective method for depolymerizing crude fucoidan from Fucus Vesiculosus seaweed.

Investigation into the use of novel pretreatments in the fermentation of Alaria esculenta by Lactiplantibacillus plantarum and kombucha SCOBY.

High pressure processing (HPP), ultrasound probe (USP) and ultrasound bath (USB) were applied to Alaria esculenta as a fermentation pre-treatment. Seaweed was then fermented by Lactiplantibacillus plantarum (LAB) or symbiotic culture of bacteria and yeast (SCOBY). Physiochemical properties of fermented seaweed were measured. pH was significantly different (p < 0.05) across SCOBY-fermented samples with different pre-treatments but not LAB-fermented samples (p > 0.05). There was a significant difference (p < 0.05) in total viable count (TVC) with the highest count in HPP-treated samples, and lowest in control samples. Organic acids differed significantly (p < 0.05) across pre-treatments for both fermentation groups. 27 volatile compounds were detected in the samples, with alcohols and ketones the most prominent groups. The quantity of volatile compounds was not significantly lower (p > 0.05) from seaweed powder. The control sample had the highest levels of tropomyosin (15.92 mg/kg) followed by HPP samples.

Added insult to injury? The response of meat-associated pathogens to proposed antimicrobial interventions.

Modern requirements for 'green label' meat products have led to the design of novel antimicrobial innovations which prioritise quality, safety and longevity. Plasma-functionalised water (PFW), ultraviolet light and natural antimicrobial compositions have been investigated and optimised for control of foodborne pathogens like Campylobacter jejuni and Salmonella enterica serovar Typhimurium. However, given the adaptive mechanisms present in bacteria under external stresses, it is imperative to understand the effect that sublethal treatment may have on the bacterial transcriptome. In this study, Salmonella Typhimurium and C. jejuni were treated with sublethal doses of ultraviolet light, a citrus juice/essential oil marinade, and 'spark' or 'glow' cold plasma generation system-produced PFW. Immediately after treatment, cells were lysed and RNA was extracted and purified. mRNA was converted to cDNA by reverse transcription-PCR and sequenced by an Illumina MiSeq® system. Sequences were filtered and analysed using the Tuxedo workflow. Sublethal treatment of Campylobacter jejuni and Salmonella Typhimurium led to increased immediate cellular and metabolic activity, as well as diversification in protein and metabolic functioning. There was further expression of pathogenesis and virulence-associated traits associated with spark PFW and marinade treatment of Salmonella Typhimurium. However, similar concerns were not raised with glow PFW or UV-treated samples. This study provides science-based evidence of the efficacy of multi-hurdle antimicrobial system using green-label marinades and PFW or UV to inactivate pathogens without upregulating virulence traits in surviving cells. This study will inform policymakers and food industry stakeholders and reinforces the need to incorporate in-line novel technologies to ensure consumer safety. KEY POINTS: • Salmonella and C. jejuni showed increased cell activity in immediate response to stress. • Virulence genes showed increased expression when treated with natural antimicrobials and sPFW. • Reduced immediate transcriptomic response to gPFW and UV treatment indicates lower risk.

Circular economy approaches for the production of high-value polysaccharides from microalgal biomass grown on industrial fish processing wastewater: A review.

The discharge of high-strength wastewater from the fish-processing industries, comprising undefined blends of toxic and organic compounds, has always been a subject of great disquiet worldwide. Despite a large number of effluent treatment methodologies known to date, biosorption with the aid of naturally grown microalgae has been recognized recently to possess promising outcomes in eradicating pollutants comprising organic compounds from liquid effluents. Interestingly, the microalgal biomass harvested from phytoremediation of fish effluent was identified to be abundant in bio compounds that exhibited potential application in pharmaceutical, nutraceutical, and, aquaculture feed, generating a circular economy. In this context, the focus of the review is to emphasize the applications of microalgal species as naturally occurring and zero-cost adsorbents for the elimination of organic contaminants from fish liquid effluents. The summary of the literature encompassed in this work is supposed to benefit the readers to comprehend the primary mechanisms by which microalgae uptakes the organic matter from fish processing effluents and converts them into various biological molecules. From the scientific works assessed through this review, the most promising microalgae species regards to nutrient uptake and removal efficiency from fish effluent, were identified as Chlorella sp. > Spirulina sp. > Scenedesmus sp. The review further revealed supercritical fluid extraction as the robust extraction tool for the extraction of targeted bioproducts from microalgal biomass grown within fish effluents. Eventually, the information presented through this review establishes phytoremediation using microalgal biomass to be a natural cost-effective, sustainable circular bio-economy approach that could be robustly applied for the efficient treatment of wastewater discharged from food processing industries.

Sequential extraction of fucoidan, laminarin, mannitol, alginate and protein from brown macroalgae Ascophyllum nodosum and Fucus vesiculosus.

The study involves development of a green biorefinery process for obtaining fucoidan, laminarin, mannitol, alginate and protein from dry and fresh Fucus vesiculosus and Ascophyllum nodosum using hydrochloric acid and a green extraction solvent. After the extraction of fucoidan which was the targeted biomolecule, an extract and by-product (residual biomass) were obtained. The extract was passed through an ultrafiltration membrane, where fucoidan was obtained in the ultrafiltration retentate while ultrafiltration permeate was analysed for laminarin and mannitol. The residual biomass was used for obtaining alginate using ultrasound (20 kHz, 64 % amplitude and 32 min, optimum parameters for alginate extraction based on our previous study). All the samples, showed good results for alginate, laminarin and mannitol, indicating that the by-products can be utilised using this green extraction process. The comparison of both dry and fresh seaweed is relevant from an industry perspective, as fresh seaweed can directly be used for extraction, avoiding drying which adds significantly to the cost of the process. Life cycle impact assessment of the complete seaweed value chain has been carried out to identify the energy demand and key environmental hotspots. This biorefinery process can be used by industry to improve their processes and utilise the by-products generated efficiently.

Ultrasound, microwave and enzyme-assisted multiproduct biorefinery of Ascophyllum nodosum.

This study investigated the multiproduct (fucoidans, ß-glucans, proteins, carotenoids, fatty acids, amino acids and polyphenols) valorization of the invasive macroalgae Ascophyllum nodosum within a green biorefinery concept using ultrasound (US), microwave (MW) treatment followed by supercritical CO2 (SC-CO2) with co-solvent and enzymatic extraction. Water and 50% aqueous ethanol were used as green extraction solvents. The extraction methods using 50% ethanol as extraction solvent improved the yields of phenolic compounds and glucan and enhanced in vitro antioxidant activity. The characterization of SC-CO2 extracts revealed that pretreatment with US and MW improved the 2-fold yield of carotenoids, total phenolics and fatty acids. However, US/MW pretreatment and enzymatic extraction did not improve the yields of proteins and free amino acids. Overall, using concurrent green US/MW-assisted extraction methods enhanced the yields of the bioactive compounds in a short duration and provided extracts with a better antioxidant capacity in the field of food applications.

Anti-methanogenic potential of seaweeds and seaweed-derived compounds in ruminant feed: current perspectives, risks and future prospects.

With methane emissions from ruminant agriculture contributing 17% of total methane emissions worldwide, there is increasing urgency to develop strategies to reduce greenhouse gas emissions in this sector. One of the proposed strategies is ruminant feed intervention studies focused on the inclusion of anti-methanogenic compounds which are those capable of interacting with the rumen microbiome, reducing the capacity of ruminal microorganisms to produce methane. Recently, seaweeds have been investigated for their ability to reduce methane in ruminants in vitro and in vivo, with the greatest methane abatement reported when using the red seaweed Asparagopsis taxiformis (attributed to the bromoform content of this species). From the literature analysis in this study, levels of up to 99% reduction in ruminant methane emissions have been reported from inclusion of this seaweed in animal feed, although further in vivo and microbiome studies are required to confirm these results as other reports showed no effect on methane emission resulting from the inclusion of seaweed to basal feed. This review explores the current state of research aiming to integrate seaweeds as anti-methanogenic feed additives, as well as examining the specific bioactive compounds within seaweeds that are likely to be related to these effects. The effects of the inclusion of seaweeds on the ruminal microbiome are also reviewed, as well as the future challenges when considering the large-scale inclusion of seaweeds into ruminant diets as anti-methanogenic agents.

A review of alternative proteins for vegan diets: Sources, physico-chemical properties, nutritional equivalency, and consumer acceptance.

Alternate proteins are gaining popularity as a more sustainable and environmentally friendly alternative to animal-based proteins. These proteins are often considered healthier and are suitable for people following a vegetarian or vegan diet. Alternative proteins can be recovered from natural sources like legumes, grains, nuts, and seeds, while single cell proteins (mycoproteins), and algal proteins are being developed using cutting-edge technology to grow fungus, yeast and algal cells in a controlled environment, creating a more sustainable source of protein. Although, the demand for alternative protein products is increasing, there still happens to be a large gap in use among the general consumers mainly stemming from its lower bioavailability, lack of nutritional equivalency and reduced digestibility compared to animal proteins. The focus of the review is to emphasize on various sources and technologies for recovering alternative proteins for vegan diets. The review discusses physicochemical properties of alternative proteins and emphasise on the role of various processing technologies that can change the digestibility and bioavailability of these proteins. It further accentuates the nutritional equivalency and environmental sustainability of alternative protein against the conventional proteins from animals. The food laws surrounding alternative proteins as well as the commercial potential and consumer acceptance of alternative protein products are also highlighted. Finally, key challenges to improve the consumer acceptability and market value of plant-based proteins would be in achieving nutrient equivalency and enhance bioavailability and digestibility while maintaining the same physicochemical properties, taste, texture, as animal proteins, has also been highlighted.

The efficiency of UV light-emitting diodes (UV-LED) in decontaminating Campylobacter and Salmonella and natural microbiota in chicken breast, compared to a UV pilot-plant scale device.

This study investigated the combined effect of Ultraviolet (UV) light-emitting diode (LED) technology treatment with refrigerated storage of chicken breast meat over 7 days on Campylobacter jejuni, Salmonella enterica serovar Typhimurium, total viable counts (TVC) and total Enterobacteriaceae counts (TEC). An optimised UV-LED treatment at 280 nm for 6 min decreased inoculated S. Typhimurium and C. jejuni populations by 0.6-0.64 log CFU/g, and TVC and TEC population by 1-1.2 log CFU/g in chicken samples. During a 7-day storage at 4 °C, a 0.73 log reduction in C. jejuni was achieved compared with non-treated samples. Moreover, the UV-LED effectiveness to reduce TVC and TEC during refrigerated storage was compared with a conventional UV lamp and a similar efficiency was observed. The impact of UV-LED and UV lamp devices on the microbial community composition of chicken meat during storage was further examined using 16 S rRNA gene amplicon sequencing. Although similar bacterial reductions were observed for both technologies, the microbial communities were impacted differently. Treatment with the UV conventional lamp increased the proportion of Brochothrix spp. In meat samples, whilst Photobacterium spp. Levels were reduced.

Bioactivity of Fucoidan-Rich Extracts from Fucus vesiculosus against Rotavirus and Foodborne Pathogens.

Marine algae are sources of bioactive components with defensive properties of great value against microbial infections. This study investigated the bioactivity of extracts from brown algae Fucus vesiculosus against rotavirus, the worldwide leading cause of acute gastroenteritis in infants and young children. Moreover, one of the extracts was tested against four foodborne bacteria: Campylobacter jejuni, Escherichia coli, Salmonella Typhimurium, and Listeria monocytogenes, and the non-pathogenic: E. coli K12. In vitro tests using MA104 cells revealed that both whole algae extracts and crude fucoidan precipitates neutralized rotavirus in a dose-responsive manner. The maximum neutralization activity was observed when the rotavirus was incubated with 100 µg mL-1 of the hydrochloric acid-obtained crude fucoidan (91.8%), although crude fucoidan extracted using citric acid also demonstrated high values (89.5%) at the same concentration. Furthermore, molecular weight fractionation of extracts decreased their antirotaviral activity and high molecular weight fractions exhibited higher activity compared to those of lower molecular weight. A seaweed extract with high antirotaviral activity was also found to inhibit the growth of C. jejuni, S. Typhimurium, and L. monocytogenes at a concentration of 0.2 mg mL-1. Overall, this study expands the current knowledge regarding the antimicrobial mechanisms of action of extracts from F. vesiculosus.

Recovery of Protein from Industrial Hemp Waste (Cannabis sativa, L.) Using High-Pressure Processing and Ultrasound Technologies.

Hemp seeds are currently used mainly for oil extraction, generating waste that could be potentially exploited further as a source of proteins and other bioactives. This study aims to valorise hemp waste (Cannabis sativa, L.) from previous oil extraction as a source of protein by analysing the effect of high-pressure processing (HPP) pre-treatments (0-600 MPa; 4-8 min) combined with conventional or ultrasound-assisted extraction (UAE) methods on protein recovery/purity, amino acid composition, and protein structure. Overall, maximum protein recovery (≈62%) was achieved with HPP (200 MPa, 8 min) with UAE. The highest protein purity (≈76%) was achieved with HPP (200 MPa, 4 min) with UAE. Overall, UAE improved the extraction of all amino acids compared to conventional extraction independently of HPP pre-treatments. Arg/Lys ratios of the protein isolates ranged between 3.78 and 5.34, higher than other vegetable protein sources. SDS-PAGE did not show visible differences amongst the protein isolates. These results seem to indicate the advantages of the use of UAE for protein recovery in the food industry and the need for further studies to optimise HPP/UAE for an accurate estimation of processing costs and their effects on the composition and structure of proteins to contribute further to the circular economy.

Investigation of differences in susceptibility of Campylobacter jejuni strains to UV light-emitting diode (UV-LED) technology.

Campylobacter jejuni remains a high priority in public health worldwide. Ultraviolet light emitting-diode technology (UV-LED) is currently being explored to reduce Campylobacter levels in foods. However, challenges such as differences in species and strain susceptibilities, effects of repeated UV-treatments on the bacterial genome and the potential to promote antimicrobial cross-protection or induce biofilm formation have arisen. We investigated the susceptibility of eight C. jejuni clinical and farm isolates to UV-LED exposure. UV light at 280 nm induced different inactivation kinetics among strains, of which three showed reductions greater than 1.62 log CFU/mL, while one strain was particularly resistant to UV light with a maximum reduction of 0.39 log CFU/mL. However, inactivation was reduced by 0.46-1.03 log CFU/mL in these three strains and increased to 1.20 log CFU/mL in the resistant isolate after two repeated-UV cycles. Genomic changes related to UV light exposure were analysed using WGS. C. jejuni strains with altered phenotypic responses following UV exposure were also found to have changes in biofilm formation and susceptibility to ethanol and surface cleaners.