Aflatoxins and fumonisins co-contamination effects on laying hens and use of mycotoxin detoxifiers as a mitigation strategy.

This study examined the effects of fumonisins (FBs) and aflatoxin B1 (AFB1), alone or in combination, on the productivity and health of laying hens, as well as the transfer of aflatoxins (AFs) to chicken food products. The efficacy and safety of mycotoxin detoxifiers (bentonite and fumonisin esterase) to mitigate these effects were also assessed. Laying hens (400) were divided into 20 groups and fed a control, moderate (54.6 µg/kg feed) or high (546 µg/kg feed) AFB1 or FBs (7.9 mg/kg feed) added diets, either alone or in combination, with the mycotoxin detoxifiers added in selected diets. Productivity was evaluated by feed intake, egg weight, egg production, and feed conversion ratio whereas health was assessed by organ weights, blood biochemistry, and mortality. Aflatoxins residues in plasma, liver, muscle, and eggs were determined using UHPLC-MS/MS methods. A diet with AFB1 at a concentration of 546 µg/kg feed decreased egg production and various AFB1-contaminated diets increased serum uric acid levels and weights of liver, spleen, heart, and gizzard. Interactions between AFB1 and FBs significantly impacted spleen, heart, and gizzard weights as well as AFB1 residues in eggs. Maximum AFB1 residues of 0.64 µg/kg and aflatoxin M1 (below limits of quantification) were observed in liver, plasma, and eggs of layers fed diets with AFB1. The mycotoxin detoxifiers reduced effects of AFB1 and FBs on egg production, organ weights, blood biochemistry, and AFB1 residues in tissues. This study highlights the importance of mycotoxin detoxifiers as a mitigation strategy against mycotoxins in poultry production.

Farmed crickets (Orthoptera: Gryllidae) raised with dermestids (Coleoptera: Dermestidae) suffer from reduced and delayed growth, but not enough to explain reports of dramatic yield loss.

The mass production of insects for food and feed is an expanding North American industry. Facilities that mass rear insects are at risk of pest infestations because the optimal environmental conditions for rearing beneficial species may also support the development of pest species. Here, we present the first recorded results detailing the interactions between dermestids and farmed crickets. Cricket farms have reported extremely low harvest yield during heavy dermestid infestations, but the exact reasons for this low yield are unknown. Many dermestid larvae are covered in dense, detachable, barbed setae called hastisetae, which are used by the larvae as an active trapping system against arthropod predators. We designed a series of experiments to test the hypotheses that a dermestid pest of cricket farms, black larder beetle (Dermestes ater DeGeer (Coleoptera: Dermestidae)), may be directly impacting Gryllodes sigillatus Walker (Orthoptera: Gryllidae) yield through the physical effects of hastisetae ingestion and/or indirectly impacting cricket yield through competition for fishmeal, a primary source of protein in conventional cricket feed. Our predictions that G. sigillatus life history and survival would be negatively affected by dermestids were largely refuted. Females fed infested diets grew less mass, but not smaller body size, compared to females fed uninfested diets. We also found that while G. sigillatus experienced delayed growth early in life after living with dermestids, they were able to tolerate living with, and consuming, dermestid larvae. We discuss how these findings have led to new hypotheses concerning how dermestid infestations drive reductions in cricket farm yield.

Mycotoxin management: exploring natural solutions for mycotoxin prevention and detoxification in food and feed.

Mycotoxins, secondary metabolites produced by various fungi, pose a significant threat to food and feed safety worldwide due to their toxic effects on human and animal health. Traditional methods of mycotoxin management often involve chemical treatments, which may raise concerns about residual toxicity and environmental impact. In recent years, there has been growing interest in exploring natural alternatives for preventing mycotoxin contamination and detoxification. This review provides an overview of the current research on the use of natural products for mitigating mycotoxin risks in food and feed. It encompasses a wide range of natural sources, including plant-derived compounds, microbial agents, and enzymatic control. The mechanisms underlying the efficacy of these natural products in inhibiting mycotoxin synthesis, adsorbing mycotoxins, or enhancing detoxification processes are discussed. Challenges and future directions in the development and application of natural products for mycotoxin management are also addressed. Overall, this review highlights the promising role of natural products as sustainable and eco-friendly alternatives for combating mycotoxin contamination in the food and feed supply chain.

Whole-genome sequencing of two captive black soldier fly populations: Implications for commercial production.

Black soldier fly (BSF; Hermetia illucens) is a promising insect species for food and feed production as its larvae can convert different organic waste to high-value protein. Selective breeding is one way to optimize production, but the potential of breeding is only starting to be explored and not yet utilized for BSF. To assist in monitoring a captive population and implementing a breeding program, genomics tools are imperative. We conducted whole genome sequencing of two captive populations separated by geographical distance - Denmark (DK) and Texas, USA (TX). Various population genetics analyses revealed a moderate genetic differentiation between two populations. Moreover, we observed higher inbreeding in the DK population, and the detection of a subpopulation within DK population aligned well with the recent foundation of the DK population from two captive populations. Additionally, we generated gene ontology annotation and variants annotation for wider potential applications. Our findings establish a robust marker set for research in population genetics, facilitating the monitoring of inbreeding and laying the groundwork for practical breeding programs for BSF.

Analysis of the original notifications in the Tuscany region "Rapid Alert System for Food and Feed" in the seven-year period 2015-2021.

The original notifications (n=1355) managed by the Rapid Alert System for Food and Feed of the Tuscany region in the seven-year period 2015-2021 were analyzed. 68.9% of them were classified as alert notifications, and they mostly originated (56.3%) from official controls on the market, followed by the company's own checks (29.1%). Italy was the most represented notifying country (73.3%) and the most represented country of origin of the notified products (64.5%). Out of the 28 considered food categories, "fish and products thereof" (F), "cereals and bakery products" (C), "bivalve mollusks and products thereof" (B), and "meat and meat products (other than poultry)" (M) were the most notified. F were especially notified for the presence of heavy metals (45.7%), C for undeclared allergens (32.1%), B for microbial contaminants (Escherichia coli, 49.6%), and M for pathogenic microorganisms (55.9%), mostly Salmonella spp. and Listeria monocytogenes. This study provides an updated framework for the main food safety issues at the regional level. The Rapid Alert System for Food and Feed should be periodically analyzed to evaluate historical trends and emerging or poorly known hazards. This is especially important for the planning of official control activities and the prevention of risks through the food supply chain.

Modernizing and harmonizing regulatory data requirements for genetically modified crops-perspectives from a workshop.

Genetically modified (GM) crops that have been engineered to express transgenes have been in commercial use since 1995 and are annually grown on 200 million hectares globally. These crops have provided documented benefits to food security, rural economies, and the environment, with no substantiated case of food, feed, or environmental harm attributable to cultivation or consumption. Despite this extensive history of advantages and safety, the level of regulatory scrutiny has continually increased, placing undue burdens on regulators, developers, and society, while reinforcing consumer distrust of the technology. CropLife International held a workshop at the 16th International Society of Biosafety Research (ISBR) Symposium to examine the scientific basis for modernizing global regulatory frameworks for GM crops. Participants represented a spectrum of global stakeholders, including academic researchers, GM crop developers, regulatory consultants, and regulators. Concurrently examining the considerations of food and feed safety, along with environmental safety, for GM crops, the workshop presented recommendations for a core set of data that should always be considered, and supplementary (i.e., conditional) data that would be warranted only on a case-by-case basis to address specific plausible hypotheses of harm. Then, using a case-study involving a hypothetical GM maize event expressing two familiar traits (insect protection and herbicide tolerance), participants were asked to consider these recommendations and discuss if any additional data might be warranted to support a science-based risk assessment or for regulatory decision-making. The discussions during the workshop highlighted that the set of data to address the food, feed, and environmental safety of the hypothetical GM maize, in relation to a conventional comparator, could be modernized compared to current global regulatory requirements. If these scientific approaches to modernize data packages for GM crop regulation were adopted globally, GM crops could be commercialized in a more timely manner, thereby enabling development of more diverse GM traits to benefit growers, consumers, and the environment.

Rearing methods of four insect species intended as feed, food, and food ingredients: a review.

Over the past 2 decades, the potential of insects as food and feed has been recognized globally. Insects as feed ingredients can improve sustainability because of their lower greenhouse gas emissions and their potential to transform organic wastes into high-quality feed rich in nutrients. However, currently, the practical use of insects as food or feed is limited by the high costs of insect production. A great deal of effort is required to improve the rearing technology necessary to establish the principles of insect farming. Several insect species have become industrialized using existing methods of production. The most common industrialized insect species intended as feed and food include the yellow mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae); the house cricket, Acheta domesticus L. (Orthoptera: Gryllidae); the black soldier fly, Hermetia illucens L. (Diptera: Stratiomyidae); and the house fly, Musca domestica L. (Diptera: Muscidae). This review focuses on describing the existing rearing methods for these 4 insect species, which may provide a basis for future research to enhance insect production capabilities.

Grinding as a slaughter method for farmed black soldier fly (Hermetia illucens) larvae: Empirical recommendations to achieve instantaneous killing.

At least 200 billion black soldier fly (Hermetia illucens) larvae (BSFL) are reared each year as food and feed, and the insect farming industry is projected to grow rapidly. Despite interest by consumers, producers, and legislators, no empirical evidence exists to guide producers in practicing humane - or instantaneous - slaughter for these novel mini-livestock. BSFL may be slaughtered via freezing, boiling, grinding, or other methods; however standard operating procedures (SOPs) and equipment design may affect the likelihood of instantaneous death using these methods. We tested how larval body size and particle size plate hole diameter affect the likelihood of instantaneous death for black soldier fly larvae that are slaughtered using a standard meat grinder. Larval body size did not affect the likelihood of instantaneous death for larvae that are 106-175 mg in mass. However, particle size plate hole diameter had a significant effect on the likelihood of instantaneous death, with only 54% of larvae experiencing an instant death when using the largest particle size plate (12-mm hole diameter) compared to 84% using the smallest particle size plate (2.55 mm). However, a higher percentage of instantaneous death (up to 99%) could be achieved by reducing the proportion of larvae that become stuck in the machine. We conclude by outlining specific recommendations to support producers in achieving a 99% instantaneous death rate through specific SOPs to be used with similarly designed machines. We also develop a protocol for producers that wish to test their own grinding SOPs.

Valorization of non-edible fruit seeds into valuable products: A sustainable approach towards circular bioeconomy.

Global imperatives have recently shown a paradigm shift in the prevailing resource utilization model from a linear approach to a circular bioeconomy. The primary goal of the circular bioeconomy model is to minimize waste by effective re-usage of organic waste and efficient nutrient recycling. In essence, circular bioeconomy integrates the fundamental concept of circular economy, which strives to offer sustainable goods and services by leveraging biological resources and processes. Notably, the circular bioeconomy differs from conventional waste recycling by prioritizing the safeguarding and restoration of production ecosystems, focusing on harnessing renewable biological resources and their associated waste streams to produce value-added products like food, animal feed, and bioenergy. Amidst these sustainability efforts, fruit seeds are getting considerable attention, which were previously overlooked and commonly discarded but were known to comprise diverse chemicals with significant industrial applications, not limited to cosmetics and pharmaceutical industries. While, polyphenols in these seeds offer extensive health benefits, the inadequate conversion of fruit waste into valuable products poses substantial environmental challenges and resource wastage. This review aims to comprehend the known information about the application of non-edible fruit seeds for synthesising metallic nanoparticles, carbon dots, biochar, biosorbent, and biodiesel. Further, this review sheds light on the potential use of these seeds as functional foods and feed ingredients; it also comprehends the safety aspects associated with their utilization. Overall, this review aims to provide a roadmap for harnessing the potential of non-edible fruit seeds by adhering to the principles of a sustainable circular bioeconomy.

Pilot market surveillance of GMM contaminations in alpha-amylase food enzyme products: A detection strategy strengthened by a newly developed qPCR method targeting a GM Bacillus licheniformis producing alpha-amylase.

Using high-throughput metagenomics on commercial microbial fermentation products, DNA from a new unauthorized genetically modified microorganism (GMM), namely the GM B. licheniformis strain producing alpha-amylase (GMM alpha-amylase2), was recently discovered and characterized. On this basis, a new qPCR method targeting an unnatural association of sequences specific to the GMM alpha-amylase2 strain was designed and developed in this study, allowing to strengthen the current GMM detection strategy. The performance of the newly developed qPCR method was assessed for its specificity and sensitivity to comply with the minimum performance requirements established by the European Network of GMO Laboratories for GMO analysis. Moreover, the transferability of the in house validated qPCR method was demonstrated. Finally, its applicability was confirmed by a pilot market surveillance of GMM contaminations conducted for the first time on 40 alpha-amylase food enzyme products labelled as containing alpha-amylase. This pilot market surveillance allowed also to highlight numerous contaminations with GMM alpha-amylase2, including frequent cross-contaminations with other GMM strains previously characterized. In addition, the presence of full-length AMR genes, raising health concerns, was also reported.

The Genome of the Yellow Mealworm, Tenebrio molitor: It's Bigger Than You Think.

BACKGROUND: Insects are a sustainable source of protein for human food and animal feed. We present a genome assembly, CRISPR gene editing, and life stage-specific transcriptomes for the yellow mealworm, Tenebrio molitor, one of the most intensively farmed insects worldwide. METHODS: Long and short reads and long-range data were obtained from a T. molitor male pupa. Sequencing transcripts from 12 T. molitor life stages resulted in 279 million reads for gene prediction and genetic engineering. A unique plasmid delivery system containing guide RNAs targeting the eye color gene vermilion flanking the muscle actin gene promoter and EGFP marker was used in CRISPR/Cas9 transformation. RESULTS: The assembly is approximately 53% of the genome size of 756.8 ± 9.6 Mb, measured using flow cytometry. Assembly was complicated by a satellitome of at least 11 highly conserved satDNAs occupying 28% of the genome. The injection of the plasmid into embryos resulted in knock-out of Tm vermilion and knock-in of EGFP. CONCLUSIONS: The genome of T. molitor is longer than current assemblies (including ours) due to a substantial amount (26.5%) of only one highly abundant satellite DNA sequence. Genetic sequences and transformation tools for an insect important to the food and feed industries will promote the sustainable utilization of mealworms and other farmed insects.

Fatty acids derived from oviposition systems guide female black soldier flies (Hermetia illucens) toward egg deposition sites.

The black soldier fly, Hermetia illucens, comes with big promises for industrial purposes since its larvae feed polyphagously on a broad spectrum of organic substrates. However, research focusing on adult flies is scarce, which is inconsistent with their reproductive relevance within the rearing cycle. In particular, directed oviposition is a challenge in artificial systems. Currently, decomposing organic matter is commonly used as oviposition substrate, which has extensive potential for improvement in view of the lack of standardization and the risk of microbial contamination. Here, we identified three fatty acids and one fatty acid methyl ester derived from the surface of old oviposition sites and targeted to elucidate their effect on preference behavior and oviposition site selection using Y-olfactometry and prepared oviposition sites, respectively. Exposure to tetradecanoic acid attracted gravid females and stimulated oviposition most strongly, while decanoic acid demonstrated a repulsive effect. Females kept in mixed-sex populations were attracted by tetradecanoic acid, resulting in a higher egg mass found in the compound box (3.0-11.4 fold), a ≥ 2.3 fold reduction of nonspecifically deposited eggs, and the highest total egg mass. Conversely, decanoic and dodecanoic acid caused females to lay a greater proportion of eggs nonspecifically outside both boxes. Our data suggest that fatty acids, especially tetradecanoic acid, are important cues for oviposition site selection in black soldier flies. In order to achieve a directed oviposition behavior, the role of further short- and long-chain fatty acids as attractants should be examined.

Mating compatibility and offspring traits evaluation among different strains of Tenebrio molitor.

The fast-growing sector of insects for food and feed stimulates researchers and the industry to explore uncharted territories, such as insect breeding, to improve economically important insect fitness traits. The yellow mealworm, Tenebrio molitor L., is one of the most thoroughly studied insect species as food and feed. However, data on mating compatibility and the effect of cross-breeding between different strains on the performance and fitness of the hybrids are scarce. In the present study, we comparatively evaluated the mating compatibility between two T. molitor inbred strains (Greek and Italian) and their outbred strains, i.e., Italian (♀)-Greek (♂) and Greek (♀)-Italian (♂), as well as the performance of their hybrid offspring. Based on the results, there is good mating compatibility among adults of the strains tested. Offspring performance, quantified as larval survival and final larval weight, followed a similar pattern among the crossings examined. Even though differences were insignificant, the outbreeding of females of the Italian strain resulted in a higher cumulative number of eggs and hatching rate and higher offspring weight. The present study sheds light on the mating compatibility of different T. molitor strains and their hybrids' economically important life table characteristics to take the mass production of insects one step further.

A comparative review on methods of detection and quantification of mycotoxins in solid food and feed: a focus on cereals and nuts.

Many emerging factors and circumstances urge the need to develop and optimize the detection and quantification techniques of mycotoxins in solid food and feed. The diversity of mycotoxins, which have different properties and affinities, makes the standardization of the analytical procedures and the adoption of a single protocol that covers the attributes of all mycotoxins a tedious or even an impossible mission. Several modifications and improvements have been undergone in order to optimize the performance of these methods including the extraction solvents, the extraction methods, the clean-up procedures, and the analytical techniques. The techniques range from the rapid screening methods, which lack sensitivity and specificity such as TLC, to a spectrum of more advanced protocols, namely, ELISA, HPLC, and GC-MS and LC-MS/MS. This review aims at assessing the current studies related to these analytical techniques of mycotoxins in solid food and feed. It discusses and evaluates, through a critical approach, various sample treatment techniques, and provides an in-depth examination of different mycotoxin detection methods. Furthermore, it includes a comparison of their actual accuracy and a thorough analysis of the observed benefits and drawbacks.

Effects of Temperature and Density on House Cricket Survival and Growth and on the Prevalence of Acheta Domesticus Densovirus.

The house cricket, Acheta domesticus, is a commonly reared insect for food and feed purposes. In 1977, a report described a colony collapse, which was caused by the single-stranded DNA virus Acheta domesticus densovirus (AdDV). Currently, there are no confirmed A. domesticus colonies free of AdDV, and viral disease outbreaks are a continuous threat to A. domesticus mass rearing. Correlations between cricket rearing density or temperature and AdDV abundance have been hypothesized, but experimental evidence is lacking. Optimised rearing conditions, including temperature and density, are key to cost-effective cricket production. In this study, house crickets were subjected to different combinations of rearing density (10, 20, 40 crickets per box) and temperature (25, 30, 35 °C) to study the effect on cricket survival, biomass, and AdDV abundance. Rearing temperature affected had a minor effect on survival, which ranged between 80 and 83%. Total cricket biomass increased with higher temperatures and higher densities. Viral abundance in crickets at the end of the rearing period was variable; however, high rearing density seemed to result in higher AdDV abundance. At 35 °C, a temperature considered suboptimal for house cricket production, viral abundance tended to be lower than at 25 or 30 °C.

Genome and Genetic Engineering of the House Cricket (Acheta domesticus): A Resource for Sustainable Agriculture.

Background: The house cricket, Acheta domesticus, is one of the most farmed insects worldwide and the foundation of an emerging industry using insects as a sustainable food source. Edible insects present a promising alternative for protein production amid a plethora of reports on climate change and biodiversity loss largely driven by agriculture. As with other crops, genetic resources are needed to improve crickets for food and other applications. Methods: We present the first high quality annotated genome assembly of A. domesticus from long read data and scaffolded to chromosome level, providing information needed for genetic manipulation. Results: Gene groups related to immunity were annotated and will be useful for improving value to insect farmers. Metagenome scaffolds in the A. domesticus assembly, including Invertebrate Iridescent Virus 6 (IIV6), were submitted as host-associated sequences. We demonstrate both CRISPR/Cas9-mediated knock-in and knock-out of A. domesticus and discuss implications for the food, pharmaceutical, and other industries. RNAi was demonstrated to disrupt the function of the vermilion eye-color gene producing a useful white-eye biomarker phenotype. Conclusions: We are utilizing these data to develop technologies for downstream commercial applications, including more nutritious and disease-resistant crickets, as well as lines producing valuable bioproducts, such as vaccines and antibiotics.

Targeted High-Throughput Sequencing Enables the Detection of Single Nucleotide Variations in CRISPR/Cas9 Gene-Edited Organisms.

Similar to genetically modified organisms (GMOs) produced by classical genetic engineering, gene-edited (GE) organisms and their derived food/feed products commercialized on the European Union market fall within the scope of European Union Directive 2001/18/EC. Consequently, their control in the food/feed chain by GMO enforcement laboratories is required by the competent authorities to guarantee food/feed safety and traceability (2003/1829/EC; 2003/1830/EC). However, their detection is potentially challenging at both the analytical and interpretation levels since this requires methodological approaches that can target and detect a specific single nucleotide variation (SNV) introduced into a GE organism. In this study, we propose a targeted high-throughput sequencing approach, including (i) a prior PCR-based enrichment step to amplify regions of interest, (ii) a sequencing step, and (iii) a data analysis methodology to identify SNVs of interest. To investigate if the performance of this targeted high-throughput sequencing approach is compatible with the performance criteria used in the GMO detection field, several samples containing different percentages of a GE rice line carrying a single adenosine insertion in OsMADS26 were prepared and analyzed. The SNV of interest in samples containing the GE rice line could successfully be detected, both at high and low percentages. No impact related to food processing or to the presence of other crop species was observed. The present proof-of-concept study has allowed us to deliver the first experimental-based evidence indicating that the proposed targeted high-throughput sequencing approach may constitute, in the future, a specific and sensitive tool to support the safety and traceability of the food/feed chain regarding GE plants carrying SNVs.

Seasonal variations in mercury, cadmium, lead and arsenic species in Norwegian blue mussels (Mytilus edulis L.) - Assessing the influence of biological and environmental factors.

BACKGROUND: Blue mussels (Mytilus edulis L.) can accumulate undesirable substances, including the potentially toxic elements (PTEs) cadmium (Cd), mercury, (Hg), lead (Pb), arsenic (As) and As species. In this study, the levels of PTEs and As species were determined in samples of blue mussels to assess the influence of environmental and biological factors, and evaluate the potential risk associated with blue mussels in terms of food and feed safety. METHODOLOGY: Blue mussels were collected monthly from one location in Western Norway from February 2018 to December 2018, and from April 2019 to April 2020. Samples were analyzed for PTEs using inductively coupled plasma mass spectrometry (ICP-MS), and high-performance liquid chromatography (HPLC) coupled to ICP-MS. Temperature, salinity and fluorescence (chlorophyll a) were monitored in the seawater column by STD/CTD, to assess the potential influence of these environmental factors on the PTE levels in the mussels. RESULTS: The results showed seasonal variations in the PTEs, with somewhat higher concentrations in spring and winter months. Unusually high levels of total As (101.2 mg kg-1 dw) and inorganic As (53.6 mg kg-1 dw) were observed for some of the time points. The organic As species arsenobetaine was generally the major As species (17-82% of total As) in the mussels, but also simple methylated As species and arsenosugars were detected. Principal components analysis (PCA) did not show a consistent relationship between the environmental factors and the PTE concentrations, showing contrary results for some elements for the periods studied. The condition index (CI) could explain variations in element concentration with significant correlations for Cd (r = -0.67, p = 0.009) and Pb (r = -0.62, p = 0.02 in 2019/20 and r = -0.52, p = 0.02 in 2018), whereas the correlation between As and CI was not significant (r = 0.12 in 2018, and r = -0.06 in 2019/20). Higher concentrations of iAs and arsenosugars coincided with increased signals of chlorophyll a, suggesting that phytoplankton blooms could be a source of As in the blue mussels. CONCLUSION: To our knowledge, this is the first study of As species in blue mussels collected over a time period of two years, providing an insight into the natural variations of these chemical forms in mussels. In terms of mussel as food and future feed material, concentrations of Cd, Hg and Pb were below the maximum levels (MLs) established in the EU food and feed legislation. However, levels of As and iAs in mussels at some time points exceeded the MLs for As in the feed legislation, and the margin of exposure (MOE) was low if these mussels were for human consumption, highlighting the importance of determining the chemical forms of As in feed and food.