Combined Dietary Supplementation of Tenebrio molitor Larvae and Chitosan in Growing Pigs: A Pilot Study.

Nowadays, the global animal industry faces considerable challenges in securing sufficient feed resources. Responding to consumer demands for reduced use of antibiotics in animal nutrition, better animal welfare status, and reduced impact on the environment, there is an increased urgency to develop innovative functional feeds with a reduced environmental footprint and the ability to improve meat quality and safety. In an effort to explore innovative feed ingredients for growing pig diets, the combined dietary supplementation of Tenebrio molitor larvae and chitosan was investigated. An experimental trial was performed with 48 weaned pigs (34 days of life; mixed sex) that were randomly assigned to four treatment groups (with six males and six females each): Group A (control), Group B (supplemented with T. molitor larvae at 10%), Group C (supplemented with chitosan at 0.05%), and Group D (supplemented with both ingredients at 10% and 0.05%, respectively). On the 42nd day of the experimental trial, samples of blood, feces, and carcass parts were taken for analysis. The results indicated that the insect larvae meal significantly improved (p < 0.05) overall performance, increased (p < 0.05) blood red blood cell content, increased meat phenolic content (p < 0.05), improved meat oxidative stability (p < 0.05), and affected meat fatty acid profile (p < 0.05). On the other hand, chitosan had no significant effect on overall performance (p > 0.05), but it significantly increased blood lymphocyte content (p < 0.05), affected the fecal microbiota (p < 0.05), improved meat oxidative stability (p < 0.05), increased meat phenolic content (p < 0.05), and affected meat fatty acid composition (p < 0.05) and (p < 0.05) meat color. Finally, the combined use of both T. molitor and chitosan significantly affected some important zootechnical parameters (p < 0.05), fecal microbial populations (p < 0.05), meat color (p < 0.05), and fatty acid profile (p < 0.05). Further investigation into the potential interaction between insect larvae meals and chitosan in pig diets is advised.

An mRNA-based reverse-vaccinology strategy to stimulate the immune response against Nipah virus in humans using fusion glycoproteins.

The zoonotic pathogen, Nipah virus, is considered a potential healthcare threat due to its high mortality rates and detrimental symptoms like encephalitis. Ribavirin, an antiviral drug helps in overcoming the number of casualties and reducing the mortality rate, but no long-lasting solution has been proposed yet putting global health security in jeopardy. Given the cognizance of mRNA-based vaccines as safe and efficacious preventative strategies against pathogens, the current study has utilized the reverse-vaccinology approach coupled with immunoinformatics to propose an mRNA-based vaccine candidate against the Nipah virus. To ensure the effectiveness of the vaccine candidate against all strains of Nipah and associated viruses, three fusion glycoproteins from Nipah and Hendra viruses were selected. A total of 30 potential epitopes, 10 B-cell-, 10 MHC-I-, and 10 MHC-II-specific, were screened for the construct. The finalized epitopes were highly antigenic with scores ranging from 0.75 to 1.7615 at a threshold of 0.4 for viruses and non-homologous to Homo sapiens eradicating any chance of immune tolerance. The construct, with a World population coverage of 97.2%, was structurally stable, thermostable, and hydrophilic with indices of 32.91, 93.62, and -0.002, respectively. The vaccine candidate's tertiary structure was predicted with a TM score of 0.131 and the refined model displayed superlative RAMA improvement (98.2) and MolProbity score (0.975). A quality factor of 93.5421% further validated the structural quality and stability. A prompt and stable immune response was also simulated, and the vaccine candidate was shown to eliminate from the body within the first five days of injection. Immune complexes count of 7000 mg/mL was predicted against the antigen with a small but nonsignificant danger signal, countered by the cytokines. Lastly, strong molecular interactions of the vaccine candidate with TLR-3 (331.09 kcal/mol) and TLR-4 (-333.31 kcal/mol) and molecular dynamics simulation analysis authenticated the immunogenic potential of the vaccine candidate. This vaccine candidate can serve as a foundation for future in-vitro and in-vivo trials to minimize or eradicate the diseases associated with the Nipah virus or the Henipaviral family.

Breeding Substrate Containing Distillation Residues of Mediterranean Medicinal Aromatic Plants Modulates the Effects of Tenebrio molitor as Fishmeal Substitute on Blood Signal Transduction and WBC Activation of Gilthead Seabream (Sparus aurata).

This work assesses the dietary use of two insect meals of Tenebrio molitor (TM) larvae reared in conventional (TM-10) or MAP-enriched substrates (MAP-TM-10) as fish meal replacements (10%) in the diets of gilthead seabream (Sparus aurata). Fish (n = 4500; 207.19 ± 1.47 g) were divided into three groups with triplicates: control (fed conventional diet), TM-10, and MAP-TM-10 groups. The fish were reared in floating cages for 12 weeks and the dietary effects on white blood cell activation, heat shock proteins, MAPKs, and apoptosis of the fish were evaluated. The MAP-TM-10 group exhibited the highest eosinophilic induction. Phosphorylated levels of p38 MAPK, p44/42 MAPK, HSP70, and HSP90 increased in the TM-10 and MAP-TM-10 groups. In terms of apoptosis, Bax levels were lower in the TM groups compared to the control, and the MAP-TM-10 group showed even lower levels than the TM-10 group. Bcl-2 levels increased in the TM-10 group compared to the control, and further increased in the MAP-TM-10 group. The Bax/Bcl-2 ratio, an apoptosis indicator, decreased in the TM groups, with the MAP-TM-10 group showing a further decrease compared to TM-10. These findings suggest that insects' breeding substrate being enriched with MAPs modulated the effect of TM on cellular stress and apoptosis.

Use of an Innovative Silage of Agro-Industrial Waste By-Products in Pig Nutrition: A Pilot Study of Its Effects on the Pig Gastrointestinal Microbiota.

The aim of this study was to evaluate whether dietary supplementation with an innovative silage (IS) created using 60% olive mill waste, 20% grape pomace, and 20% deproteinised feta cheese waste solids can modulate the composition of the intestinal microbiota in weaned (Exp. 1) and finishing (Exp. 2) pigs. In Exp. 1 (40 day supplementation), forty-five crossbred weaned pigs were randomly assigned to the 0% (Control), 5%, or 10% IS groups (15 replicates/experimental diet). In Exp. 2 (60 day supplementation), eighteen finishing pigs from Exp. 1 were fed the control diet for 8 weeks before being re-assigned to their original experimental groups and fed with the 0% (Control), 5%, or 10% IS diets (six replicates/experimental diet). Performance parameters were recorded. Ileal and caecal digesta and mucosa were collected at the end of each experiment for microbiota analysis using 16S rRNA gene sequencing (five pigs/experimental diet for Exp. 1 and six pigs/experimental diet for Exp. 2). No significant effects on pig growth parameters were observed in both experiments. In Exp. 1, 5% IS supplementation increased the relative abundance of the Prevotellaceae family, Coprococcus genus, and Alloprevotella rava (OTU_48) and reduced the relative abundance of Lactobacillus genus in the caecum compared to the control and/or 10% IS diets (p < 0.05). In Exp. 2, 5% IS supplementation led to compositionally more diverse and different ileal and caecal microbiota compared to the control group (p < 0.05; p = 0.066 for ß-diversity in ileum). Supplementation with the 5% IS increased the relative abundance of Clostridium celatum/disporicum/saudiense (OTU_3) in the ileum and caecum and Bifidobacterium pseudolongum (OTU_17) in the caecum and reduced the relative abundance of Streptococcus gallolyticus/alactolyticus (OTU_2) in the caecum compared to the control diet (p < 0.05). Similar effects on C. celatum/disporicum/saudiense and S. gallolyticus/alactolyticus were observed with the 10% IS diet in the caecum (p < 0.05). IS has the potential to beneficially alter the composition of the gastrointestinal microbiota in pigs.

Utilization of Tenebrio molitor Larvae Reared with Different Substrates as Feed Ingredients in Growing Pigs.

The procurement of adequate feed resources is one of the most important challenges for the animal industry worldwide. While the need for feeds rich in protein is constantly increasing, their production cannot readily keep up. Consequently, to overcome this challenge in a sustainable way, it is necessary to identify and develop new feeding strategies and feed ingredients, such as insect meals. In the present study, Tenebrio molitor larvae that were reared on two different substrates (standard and enriched with medicinal aromatic plant material) were used as feed ingredients for growing pigs. A total of 36 weaned pigs (34 days old) were randomly allocated to three treatment groups and fed either the control diet (A) or diets supplemented at 10% with one of the two insect meals (B and C). At the end of the trial (42 days), blood, feces, and meat samples were collected for analysis. The insect meal supplementation did not affect (p > 0.05) overall performance but significantly modified (p < 0.001) the fecal microflora balance and the blood cholesterol (p < 0.001), while the rest of the blood parameters tested were not affected. Moreover, this dietary supplementation significantly affected some microbial populations (p < 0.001), improved the total phenolic content (p < 0.05), and the fatty acid profile (p < 0.001) of the meat cuts, but did not affect (p > 0.05) meat color or proximate composition. Further research is needed to evaluate the different types and levels of inclusion of insect meals in pig nutrition.

Innovative Use of Olive, Winery and Cheese Waste By-Products as Novel Ingredients in Weaned Pigs Nutrition.

New sustainable sources of feeds, which can enhance the health and welfare of farm animals, lower feeding costs, and lead to safer products, are of high priority in modern animal production systems. In the present study, a novel silage created from Greek olive, winery, and feta cheese waste by-products, was evaluated as a feed ingredient at different inclusion rates (0%, 5% or 10%) in 34-day-old weaned pigs. The potential beneficial effects on performance, health and intestinal digesta microflora balance of the pigs were evaluated. Additionally, chemical, microbiological and quality analysis of the meat was carried out. Results showed no detrimental effects (p > 0.05) on the pigs' performance and no significant changes (p > 0.05) in meat pH, color and chemical analysis. Ileum and cecum microflora populations (total anaerobes, Lactobacillaceae) were positively affected (p ≤ 0.05) by the dietary usage of the silage. The microbial populations (Clostridium spp.) of belly meat cuts were positively modified (p ≤ 0.01). The concentration of total phenols in the meat cuts were increased (p ≤ 0.05) and their resistance to oxidation was improved (p ≤ 0.05). In addition, the fatty acid profile of the meat lipids (polyunsaturated and n-3 fatty acids) was positively modified (p ≤ 0.001).