Subchronic oral exposure to polystyrene microplastics affects hepatic lipid metabolism, inflammation, and oxidative balance in gilthead seabream (Sparus aurata).

Microplastics (MPs) pose a clear threat to aquatic organisms affecting their health. Their impact on liver homeostasis, as well as on the potential onset of nonalcoholic fatty liver disease (NAFLD), is still poorly investigated and remains almost unknown. The aim of this study was to evaluate the outcomes of subchronic exposure to polystyrene MPs (PS-MPs; 1-20 µm; 0, 25, or 250 mg/kg b.w./day) on lipid metabolism, inflammation, and oxidative balance in the liver of gilthead seabreams (Sparus aurata Linnaeus, 1758) exposed for 21 days via contaminated food. PS-MPs induced an up-regulation of mRNA levels of crucial genes associated with lipid synthesis and storage (i.e., PPARy, Srebp1, Fasn) without modifications of genes involved in lipid catabolism (i.e., PPARα, HL, Pla2) or transport and metabolism (Fabp1) in the liver. The increase of CSF1R and pro-inflammatory cytokines gene expression (i.e., TNF-α and IL-1ß) was also observed in exposed fish in a dose-dependent manner. These findings were confirmed by hepatic histological evaluations reporting evidence of lipid accumulation, inflammation, and necrosis. Moreover, PS-MPs caused the impairment of the hepatic antioxidant defense system through the alteration of its enzymatic (catalase, superoxide dismutase, and glutathione reductase) and non-enzymatic (glutathione) components, resulting in the increased production of reactive oxygen species (ROS) and malondialdehyde (MDA), as biomarkers of oxidative damage. The alteration of detoxifying enzymes was inferred by the decreased Ethoxyresorufin-O-deethylase (EROD) activity and the increased activity of glutathione-S-transferase (GST) at the highest PS-MP dose. The study suggests that PS-MPs affect the liver health of gilthead seabream. The liver dysfunction and damage caused by exposure to PS-MPs result from a detrimental interplay of inflammation, oxidative damage, and antioxidant and detoxifying enzymatic systems modifications, altering the gut-liver axis homeostasis. This scenario is suggestive of the involvement of MP-induced effects in the onset and progression of hepatic lipid dysfunction in gilthead seabream.

Dietary Tenebrio molitor Larvae Meal Inclusion Exerts Tissue-Specific Effects on Cellular, Metabolic, and Antioxidant Status in European Sea Bass (Dicentrarchus labrax) and Gilthead Seabream (Sparus aurata).

The present study addresses the effects of dietary Tenebrio molitor (TM) larvae meal inclusion on cytoprotective, cell death pathways, antioxidant defence, and intermediate metabolism in the heart, muscle, and digestive tract of gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax). Three experimental diets were formulated to contain 0%, 25%, or 50% inclusion TM levels. Heat Shock Proteins (HSPs) induction was apparent in both species' muscle at 50% inclusion. Conversely, p44/42 Mitogen-Activated Protein Kinase (MAPK) activation was increased (p < 0.05) in both species' muscle and digestive tract at 25% inclusion. Regarding the apoptotic machinery, TM inclusion exerted no influence on gilthead seabream, while suppression through autophagy may have occurred in the muscle. However, significant apoptosis (p < 0.05) was evident in European sea bass muscle and digestive tract. Both fish species' heart seemed to additionally rely on lipids compared to muscle and digestive tract. In contrast to gilthead seabream, European sea bass exhibited increased (p < 0.05) antioxidant activity at 50% TM inclusion. The present findings highlight the dietary derived induction of cellular responses in a species- and tissue-specific manner, whereas European sea bass appears to be more susceptible to TM inclusion.

Identification of spectral responses of different plastic materials by means of multispectral imaging.

In this work, multispectral imaging (MSI) is introduced as an innovative, practical, and non-invasive solution capable of identifying and detecting (micro)plastics. MSI holds significant appeal for industry due to its flexibility, ease of implementation, and portability. The integration of MSI with Principal Components Analysis (PCA) enables precise identification of different plastics and differentiation of microplastics within mixtures. The technique successfully identifies and quantifies the pure spectral response (endmembers) of each microplastic in every pixel of the original image. As a result, the model excels in distinguishing specific plastic materials from their surrounding backgrounds. This novel approach facilitates the identification of randomly dispersed microplastics in water.

Enhancement of fruit byproducts through bioconversion by Hermetia illucens (Diptera: Stratiomyidae).

Bioconversion is a biological process by which organic materials are converted into products with higher biological and commercial value. During its larval stage the black soldier fly Hermetia illucens is extremely voracious and can feed on a wide variety of organic materials. To study the impact of different fruit byproducts on the insect's growth, final larval biomass, substrate reduction, bioconversion parameters, and larval nutritional composition, 10 000 black soldier fly larvae (BSFL) were reared on 7.0 kg of one of three substrates (strawberry, tangerine, or orange) or on a standard diet as a control. The results highlight that BSFL can successfully feed and grow on each of these diets, though their development time, growth rate, and final biomass were differently impacted by the substrates, with strawberry being the most suitable. The lipid and protein contents of BSFL were similar among larvae fed on different substrates; however, major differences were detected in ash, micronutrient, fiber, fatty acid, and amino acid contents. Overall, the results indicate that fruit waste management through the BSFL bioconversion process represents a commercially promising resource for regional and national agrifood companies. Our study offers new perspectives for sustainable and environmentally friendly industrial development by which fruit byproducts or waste might be disposed of or unconventionally enhanced to create secondary products of high biological and economic value, including BSFL biomass as animal feed or, in perspective, as alternative protein source for human nutrition.

Subchronic Exposure to Polystyrene Microplastic Differently Affects Redox Balance in the Anterior and Posterior Intestine of Sparus aurata.

Microplastics (MPs) are pollutants widely distributed in aquatic ecosystems. MPs are introduced mainly by ingestion acting locally or in organs far from the gastroenteric tract. MPs-induced health consequences for fish species still need to be fully understood. We aimed to investigate the effects of the subchronic oral exposure to polystyrene microplastics (PS-MPs) (1-20 µm) in the gilthead seabreams (Sparus aurata) used as the experimental model. We studied the detrimental impact of PS-MPs (25 and 250 mg/kg b.w./day) on the redox balance and antioxidant status in the intestine using histological analysis and molecular techniques. The research goal was to examine the anterior (AI) and posterior intestine (PI) tracts, characterized by morphological and functional differences. PS-MPs caused an increase of reactive oxygen species and nitrosylated proteins in both tracts, as well as augmented malondialdehyde production in the PI. PS-MPs also differently affected gene expression of antioxidant enzymes (i.e., superoxide dismutase, catalase, glutathione reductase). Moreover, an increased up-regulation of protective heat shock proteins (HSPs) (i.e., hsp70 and hsp90) was observed in PI. Our findings demonstrate that PS-MPs are responsible for oxidative/nitrosative stress and alterations of detoxifying defense system responses with differences in AI and PI of gilthead seabreams.

Impact of polystyrene microplastic exposure on gilthead seabream (Sparus aurata Linnaeus, 1758): Differential inflammatory and immune response between anterior and posterior intestine.

Plastics are the most widely discharged waste into the aquatic ecosystems, where they break down into microplastics (MPs) and nanoplastics (NPs). MPs are ingested by several marine organisms, including benthic and pelagic fish species, contributing to organ damage and bioaccumulation. This study aimed to assess the effects of MPs ingestion on gut innate immunity and barrier integrity in gilthead seabreams (Sparus aurataLinnaeus, 1758) fed for 21 days with a diet enriched with polystyrene (PS-MPs; 1-20 µm; 0, 25 or 250 mg /kg b.w./die). Physiological fish growth and health status were not impacted by PS-MPs treatments at the end of experimental period. Inflammation and immune alterations were revealed by molecular analyses in both anterior (AI) and posterior intestine (PI) and were confirmed by histological evaluation. PS-MPs triggered TLR-Myd88 signaling pathway with following impairment of cytokines release. Specifically, PS-MPs increased pro-inflammatory cytokines gene expression (i.e., IL-1ß, IL-6 and COX-2) and decreased anti-inflammatory ones (i.e., IL-10). Moreover, PS-MPs also induced an increase in other immune-associated genes, such as Lys, CSF1R and ALP. TLR-Myd88 signaling pathway may also lead to the mitogen-activated protein kinases (MAPK) signaling pathway activation. Here, MAPK (i.e., p38 and ERK) were activated by PS-MPs in PI, following the disruption of intestinal epithelial integrity, as evidenced by reduced gene expression of tight junctions (i.e. ZO-1, Cldn15, Occludin, and Tricellulin), integrins (i.e., Itgb6) and mucins (i.e., Muc2-like and Muc13-like). Thus, all the obtained results suggest that the subchronic oral exposure to PS-MPs induces inflammatory and immune alterations as well as an impact on intestinal functional integrity in gilthead seabream, with a more evident effect in PI.

Epidemiology and geographical variation of myasthenia gravis in the province of Pavia, Italy.

BACKGROUND AND AIMS: Previous studies have reported a prevalence estimate of myasthenia gravis (MG) from 7.7 to 11.1 per 100,000 inhabitants in Europe. Moreover, the study of the geographical distribution of MG should be useful to generate specific hypotheses. The aims are to estimate MG prevalence and to investigate its geographical variation in a delimited area in Northern Italy. METHODS: The primary source of data was the MG database of the Neurological Institute of Pavia and all other sources of case collection in and outside the province. We adopted a Bayesian approach to analyze MG geographical variation within the finest geographical grid. RESULTS: We identified 119 live MG prevalent cases resident in the province of Pavia on December 31, 2008. The overall crude prevalence was 24 per 100,000 inhabitants. The Bayesian analysis identified a small cluster of higher MG prevalence in the northern area of the province. CONCLUSIONS: The estimated MG prevalence sets the province of Pavia among the high-risk areas. The identification of high/low MG risk areas deserves further investigation of genetic and environmental factors possibly related to a major risk of the disease in that area.

COVID-19 pandemic: impacts on bees, beekeeping, and potential role of bee products as antiviral agents and immune enhancers.

COVID-19 pandemic has passed to the front all the contradictions of the beekeeping sector: the valuable role of bee products as immune enhancers and antiviral agents and the impact that unsustainability of human activities has on bees' health and survival. The COVID-19 emergency led several countries to adopt severe restriction measures to contrast the infection. The lowering of industrial and commercial activities, transports, and the general lockdown had immediate consequences on the air quality, significantly improving environmental conditions. This had a positive impact on honeybees' life's quality. On the other hand, the bee and beehive transportation limitations threaten to hit food production by affecting the pollinator service, and this is particularly true in large, food-exporting countries like the USA and China where due to the few numbers of local bees, beekeepers import them by other countries and convey by truck hives for thousands of kilometers to pollinate crops. Furthermore, honeybee products, focusing on their natural pharmacological properties, can play an essential role as a potential natural contrast to the virus by enhancing the immunity defenses of both humans and animals, and their demand by consumers is expected to increase. Several researchers in the last months focused their attention on bee products to evaluate their effect in the cure of COVID-19 patients to ameliorate the symptoms or to contrast the coronavirus directly. This review reports these preliminary results.

Tenebrio molitor larvae meal inclusion affects hepatic proteome and apoptosis and/or autophagy of three farmed fish species.

Herein, the effect of dietary inclusion of insect (Tenebrio molitor) meal on hepatic pathways of apoptosis and autophagy in three farmed fish species, gilthead seabream (Sparus aurata), European seabass (Dicentrarchus labrax) and rainbow trout (Oncorhynchus mykiss), fed diets at 25%, 50% and 60% insect meal inclusion levels respectively, was investigated. Hepatic proteome was examined by liver protein profiles from the three fish species, obtained by two-dimensional gel electrophoresis. Although cellular stress was evident in the three teleost species following insect meal, inclusion by T. molitor, D. labrax and O. mykiss suppressed apoptosis through induction of hepatic autophagy, while in S. aurata both cellular procedures were activated. Protein abundance showed that a total of 30, 81 and 74 spots were altered significantly in seabream, European seabass and rainbow trout, respectively. Insect meal inclusion resulted in individual protein abundance changes, with less number of proteins altered in gilthead seabream compared to European seabass and rainbow trout. This is the first study demonstrating that insect meal in fish diets is causing changes in liver protein abundances. However, a species-specific response both in the above mentioned bioindicators, indicates the need to strategically manage fish meal replacement in fish diets per species.

Effects of Dietary Supplementation with Honeybee Pollen and Its Supercritical Fluid Extract on Immune Response and Fillet's Quality of Farmed Gilthead Seabream (Sparus aurata).

The awareness of the correlation between administered diet, fish health and products' quality has led to the increase in the research for innovative and functional feed ingredients. Herein, a plant-derived product rich in bioactive compounds, such as honeybee pollen (HBP), was included as raw (HBP) and as Supercritical Fluid Extracted (SFE) pollen (HBP_SFE) in the diet for gilthead seabream (Sparus aurata). The experiment was carried out on 90 fish with an average body weight of 294.7 ± 12.8 g, divided into five groups, according to the administration of five diets for 30 days: control diet (CTR); two diets containing HBP at 5% (P5) and at 10% (P10) level of inclusion; two diets containing HBP_SFE, at 0.5% (E0.5) and at 1% (E1) level of inclusion. Their effects were evaluated on 60 specimens (336.2 ± 11.4 g average final body weight) considering the fish growth, the expression of some hepatic genes involved in the inflammatory response (il-1ß, il-6 and il-8) through quantitative real-time PCR, and physico-chemical characterization (namely color, texture, water holding capacity, fatty acid profile and lipid peroxidation) of the fish fillets monitored at the beginning (day 0) and after 110 days of storage at −20 °C. The results obtained showed that the treatment with diet E1 determined the up-regulation of il-1ß, il-6, and il-8 (p < 0.05); however, this supplementation did not significantly contribute to limiting the oxidative stress. Nevertheless, no detrimental effect on color and the other physical characteristics was observed. These results suggest that a low level of HBP_SFE could be potentially utilized in aquaculture as an immunostimulant more than an antioxidant, but further investigation is necessary.

Anti-Ma2/Ta antibodies in a woman with primary lateral sclerosis-like phenotype and Sjögren syndrome.

Anti-Ma2/Ta antibodies are rare paraneoplastic antibodies, which are mostly associated with limbic encephalitis in male patients with testicular cancer. We report on a 50-year-old woman with a pure progressive spastic paraparesis. Next, she was diagnosed as having a Sjögren syndrome, with serological positivity for anti-SS-Ro antibodies. The patient's serum and cerebrospinal fluid samples were positive for anti-Ma2/Ta antibodies, which were also proved to be intrathecally produced. These findings, and the coexistence of systemic autoimmunity, led us to treat the patient with corticosteroids first, and then with plasma exchange. Neurological symptoms scarcely responded to both the therapies. The search for cancer was negative up to 4 years after the disease onset. Our case expands the spectrum of clinical syndromes associated with anti-Ma2/Ta antibodies.

Different Combinations of Butchery and Vegetable Wastes on Growth Performance, Chemical-Nutritional Characteristics and Oxidative Status of Black Soldier Fly Growing Larvae.

Hermetia illucens larvae (five days old) were farmed on broiler feed (control diet), a vegetable diet (V100), a 50% of vegetable diet + 50% of butchery wastes (V50 + B50), and a 75% of vegetable diet + 25% of butchery wastes (V75 + B25) to evaluate their suitability. Ten kilograms of substrate and 6000 larvae composed each replicate (nine per group). Larvae were weighed and measured every two days until the 25% developed into prepupae. Larval mortality and growing indexes were calculated. Substrates, larvae, and frass chemical composition were analyzed. Larvae oxidative status and stability were measured in hemolymph and body. The V100 larvae showed the lowest live weight, length, thickness, and growth rate but had low mortality rate and high substrate reduction index and protein conversion ratio. The V100 larvae had similar protein to and lower lipids than the control ones, while the V50 + B50 and V75 + B25 larvae contained higher lipids and lower protein than the others. Despite the vegetable wastes, at different levels, the reactive oxygen species content decreased in hemolymph, and the V100 diet depressed growth performance and should be avoided. The use of butchery wastes combined with vegetable ingredients can be a suitable alternative to balance the high level of lipid and the low content of protein.

The angiotensin-converting enzyme insertion/deletion polymorphism modifies the clinical outcome in patients with Pompe disease.

PURPOSE: The insertion/deletion polymorphism of angiotensin-converting enzyme may influence muscle properties. We examined whether Pompe disease clinical manifestations, which are known to be highly variable among late-onset patients, may be modulated by angiotensin-converting enzyme polymorphism. METHODS: We included 38 patients with late-onset Pompe disease, aged 44.6 +/- 19.8 years. We compared the distribution of angiotensin-converting enzyme polymorphism according to demographic and disease parameters. RESULTS: The distribution of angiotensin-converting enzyme polymorphism was in line with the general population, with 16% of patients carrying the II genotype, 37% carrying the DD genotype, and the remaining patients with the ID genotype. The three groups did not differ in mean age, disease duration, Walton score, and other scores used to measure disease severity. The DD polymorphism was associated with earlier onset of disease (P = 0.041), higher creatine kinase levels at diagnosis (P = 0.024), presence of muscle pain (P = 0.014), and more severe rate of disease progression (P = 0.037, analysis of variance test for interaction). DISCUSSION: These findings suggest a potential role of angiotensin-converting enzyme polymorphism in modulating Pompe disease phenotype and prognosis.

Blood profile and productive performance after partial substitution of maize grain with ancient wheat lines by-products in organic laying hens' diet.

The aim of this research was to evaluate the effects of the partial substitution of maize grain with local ancient wheats by-products in an organic laying hens farm on animal performance and blood profile, as well as on eggs quality. A total of 80 Hy-Line W-36 Single Comb White Leghorn, 18 weeks old hens were homogenously divided into two groups. The two groups (40 laying hens each, 8 replicates, 5 hens/replicate) were fed two isoprotein and isoenergetic diets: the Control group (C) was fed a standard, organic maize-soybean meal diet whereas in the Ancient Grains group (AG) part of the maize grain was replaced by a mix of ancient grains middling, consisting in 50:50 of Triticum aestivum var. spelta (spelt) and Triticum durum dicoccum L. (emmer wheat). The birds fed the maize based-diet (C) showed a higher (P < 0.01) egg weight and feed intake than the Ancient Grains (AG) group. By contrary, the feed conversion ratio was more favourable (P < 0.05) in hens fed the Ancient Grains diet. The eggs produced by the hens fed the Control diet showed higher length and width than the other group (P < 0.01). The shell thickness and strength were also higher in the Control group (P < 0.01 and P < 0.05, respectively). The experimental dietary treatment positively affected some blood parameters. The Control group showed higher levels of cholesterol and triglycerides (P < 0.05) as well as of alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) (P < 0.01). In addition, butyrate, which may have great implications for the regulation of the immune response, resulted significantly higher in the caecal content of hens from the Ancient Grains group. Overall, the Ancient Grains diet seemed to be able to guarantee the production performances with positive effects on the animal health.

The Inclusion of a Supercritical Fluid Extract, Obtained From Honey Bee Pollen, in the Diet of Gilthead Sea Bream (Sparus aurata), Improves Fish Immune Response by Enhancing Anti-oxidant, and Anti-bacterial Activities.

In the present study, the immune-stimulatory effect of two levels of honey bee pollen (5 and 10%, P5 and P10 treatment, respectively) and its supercritical fluid extract (0. 5 and 1%, E0.5 and E1, respectively) included in the diet, was tested in gilthead seabream (Sparus aurata). The in vivo trial was preceded by the evaluation of antioxidant properties of three different bee pollen extracts obtained by water, ethanol 80%, and Supercritic Fluids Extraction (SFE). The preliminary evaluation attested that the SFE showed the lowest extraction yield (10.47%) compared to ethanol 80% (48.61%) and water (45.99%). SFE extract showed good antioxidant properties with high polyphenol content (13.06 mg GAE/g), radical scavenging activity (3.12 mg/ml), reducing power (38.68 mg/mL EC50). On the contrary, the water extract showed the significantly lowest polyphenol content (2 mg GAE/g; P < 0.05). The results of in vivo trial demonstrate that the diets supplemented with SFE bee pollen extract had a stimulatory effect on fish serum immunity, respect to the inclusion of raw pollen, this latter revealing some inhibitory effects in the immune response, such a decrease of serum peroxidase and lysozyme activities, particularly in P10 group significantly different (P < 0.05) from the control group. On the contrary, serum peroxidase, protease, antiprotease, were significantly increased in fish fed the diets supplemented with supercritical fluid extract, respect to the fish fed on control and on diets supplemented with 5 and 10% of raw pollen. For what concerns the bactericidal activity against Vibrio harveyii, all the treatments containing bee pollen regardless of the type showed their serum bactericidal activity significantly increased with respect to the control groups (p < 0.05). Given its high antioxidant properties, the absence of toxic solvents and the positive action carried out on improving the humoral response in gilthead seam bream, honey bee pollen SFE extract can be taken into account in the formulation of fish feeds.

Honey Bee Pollen in Meagre (Argyrosomus regius) Juvenile Diets: Effects on Growth, Diet Digestibility, Intestinal Traits, and Biochemical Markers Related to Health and Stress.

This research aimed to evaluate the effects of the inclusion of honey bee pollen (HBP) in meagre (Argyrosoumus regius) juveniles' diets on growth performance, diet digestibility, intestinal morphology, and immunohistochemistry. Furthermore, stress-related molecular markers and biochemical blood profile of fish were evaluated, together with mineral trace and toxic element concentration in pollen and diets. Specimens of meagre (360) of 3.34 ± 0.14 g initial body weight, were randomly allocated to twelve 500 L circular tanks (30 fish per tank). Four diets were formulated: a control diet and three experimental diets with 1%, 2.5%, and 4% of HBP inclusion. All the growth parameters and crude protein and ether extract digestibility coefficients were negatively linearly affected by increased HBP inclusion (p < 0.0001). Histology of medium intestine showed slight signs of alterations in group HPB1 and HPB2.5 compared to control. Fish from HBP4 group showed severe alterations at the intestinal mucosa level. Immunohistochemical detection of TNF-α in the medium intestine showed the presence of TNF-α+ cells in the lamina propria, which resulted in accordance with the increased level of the TNF-α protein detected by immunoblotting in the liver. This stress situation was confirmed by the increased hepatic level of HSP70 (p < 0.05) in fish fed the HBP4 diet and by the linear decrease of total serum protein levels in HBP-containing diets (p < 0.0001). These negative effects can be related to the ultrastructure of the bee pollen grain walls, which make the bioactive substances unavailable and can irritate the intestine of a carnivorous fish such as meagre.

Mirrors Can Affect Growth Rate, Blood Profile, Carcass and Meat Traits and Caecal Microbial Activity of Rabbits Reared in a "Small Group" Free-Range System.

The aim of this work was to propose a model of free-range raising for rabbit able to maximize the animal welfare and at the same time the productive performances through the use of mirrors. A total of 81 rabbits were allocated into free-range areas and divided into three groups (nine replicates per group): in the first group (face to face, F2F), the rabbits of each replicate could see each other. In the second group (blind) each replicate was isolated from the others; in the third group (mirrors), the replicates were divided as for the Blind group but two mirrors were placed in a corner of the perimeter. The blind group rabbits showed the lowest final weight (p < 0.05), while rabbits from the mirrors groups showed the best FCR and net dressing out values. The blind group showed the highest production of total short chain fatty acids, acetate (p < 0.05) and propionate (p < 0.01). The F2F rabbits showed higher levels of creatine phosphokinase and lactate dehydrogenase and lower values of blood glucose than those of the other groups, due to the higher locomotion activity. The use of mirrors can improve rabbit's growth performance and carcass traits by lowering the rabbit's locomotion activity in comparison to the other tested systems.

Mirrors Improve Rabbit Natural Behavior in a Free-Range Breeding System.

The aim of this research was to evaluate the possible usefulness of mirrors in improving rabbit behavior in a free-range breeding system. Three groups (each consisting of nine replicates of three animals) were compared: isolated, isolated with mirrors and separated by a wire mesh (possible visual and olfactory contacts). Rabbits allowed to have a visual and olfactory contact showed a significantly higher expression of important natural behaviors (olfactory investigation, gnawing, alertness, stretching, locomotion) compared to the isolated rabbits (with or without mirrors); while rabbits in the mirror group showed higher allo-grooming activity than those isolated and no different locomotion activity than those separated by wire mesh. Thus, mirrors seemed to be able to modify the behavioral repertoire of isolated rabbits by acting on social perception in rabbits reared in small groups in a free-range system. Nevertheless, despite it being advisable to use a combination of different indicators in order to assess the stress level of an animal, the accuracy of serum cortisol, as well as of some secondary stress markers assay, appeared to be limited in this type of breeding.

Intestinal Morphometry, Enzymatic and Microbial Activity in Laying Hens Fed Different Levels of a Hermetia illucens Larvae Meal and Toxic Elements Content of the Insect Meal and Diets.

To evaluate the effects of feeding a Hermetia illucens (HI) larvae meal on the different intestinal traits of hens, and to determine the toxic elements' concentration in the insect meal and diets, 162 hens were randomly allotted to three groups. The control received a corn-soybean meal-based diet (SBM); the HI25 and HI50 groups received two diets in which the 25% and 50% of the dietary protein were replaced by the HI protein, respectively. The duodenal and jejunal villi height and villi/crypt were higher (p < 0.01) in the SBM than in the HI groups. The ileal villi height was higher (p < 0.05) in the SBM and HI25 groups than the HI50. The HI50 group exhibited a lower duodenal maltase activity. The intestinal alkaline phosphatase (IAP) activity linearly decreased in the duodenum and jejunum as the dietary insect meal inclusion increased. The HI50 group had a higher acetate and butyrate level than the SBM. The levels of cadmium (Cd), lead (Pb), mercury (Hg), and arsenic (As) in the diets and insect meal were lower than the maximum values established by the EU Commission. The 25% soybean protein replacement with Hermetia illucens larvae meal in the diet of laying hens was more suitable and closer to the optimal level than 50%.