Organoids are self-organizing, self-renewing three-dimensional cellular structures that resemble organs in structure and function. They can be derived from adult stem cells, embryonic stem cells, or induced pluripotent stem cells. They contain most of the relevant cell types with a topology and cell-to-cell interactions resembling that of the in vivo tissue. The widespread and increasing adoption of organoid-based technologies in human biomedical research is testament to their enormous potential in basic, translational- and applied-research. In a similar fashion there appear to be ample possibilities for research applications of organoids from livestock and companion animals. Furthermore, organoids as in vitro models offer a great possibility to reduce the use of experimental animals. Here, we provide an overview of studies on organoids in livestock and companion animal species, with focus on the methods developed for organoids from a variety of tissues/organs from various animal species and on the applications in veterinary research. Current limitations, and ongoing research to address these limitations, are discussed. Further, we elaborate on a number of fields of research in animal nutrition, host-microbe interactions, animal breeding and genomics, and animal biotechnology, in which organoids may have great potential as an in vitro research tool.

Black soldier fly (Hermetia illucens; BSF) larvae as dietary protein source have the ability to deliver nutrients and could possess functional properties that positively support animal productivity and health. More knowledge, however, is needed to assess the impact of feeding a BSF based diet on gut and animal health. Sixteen post-weaned male pigs were randomly assigned to two groups and fed for three weeks with iso-caloric and iso-proteinaceous experimental diets prepared with either soybean meal (SBM) as reference protein source or with BSF as single source of dietary protein. At the end of the trial, the pigs were sacrificed to collect relevant digesta, gut tissue and blood samples to study changes induced by the dietary treatments using ~ omics based analyses. Inclusion of BSF in the diet supports the development of the intestinal microbiome that could positively influence intestinal health. By amine metabolite analysis, we identified two metabolites i.e. sarcosine and methionine sulfoxide, in plasma that serve as markers for the ingestion of insect based ingredients. BSF seems to possess functional properties indicated by the appearance of alpha-aminobutyric acid and taurine in blood plasma of pigs that are known to induce health beneficial effects.

The fatty acid content of microalgae, especially the high content of omega-3 fatty acids such as eicosapentaenoic acid (EPA, C20:5) and docosahexaenoic acid (DHA, C22:6), could enrich eggs when fed to laying hens. Moreover, the properties and bioactive components of omega-3 fatty acids could positively influence the health and production performance of laying hens. In this study, the effects of dried Nannochloropsis limnetica inclusions in diets on yolk omega-3 fatty acid content, laying hen performance, nutrient retention, intestinal morphometry and systemic inflammatory markers were measured. A total of 240 twenty-five-wk-old laying hens were randomly assigned to 5 treatments distributed among 30 pens. Treatment A received the reference diet, while diets in treatments B, C, and D contained the control diet with 1, 2, and 3% microalgae added, respectively. In treatment E, a portion of ingredients of the control diet was replaced with rapeseed meal to induce a mild nutritional challenge, along with an inclusion of 3% microalgae. Compared to the control group the rate of lay increased by approximately 5% (P = 0.039) when birds were fed 2 or 3% microalgae. Furthermore, inclusion of 2 and 3% microalgae resulted in higher feed intake compared to the control group (126, 125, and 119 g/hen/d respectively; P = 0.001). Other performance parameters such as nutrient retention and egg characteristics were not affected by the dietary treatments. The EPA and DHA content of the yolk increased with increasing microalgae inclusion level (P < 0.001). A 2% algal inclusion resulted in 58.3 (EPA) and 603 (DHA) mg per 100 g dry yolk, respectively. Plasma haptoglobin levels of laying hens in both treatments receiving 3% microalgae were almost 3 times lower compared to the control group (1.25 and 1.62 vs. 5.60; P < 0.001), regardless of the inclusion of rapeseed in the diet. Based on these results, it can be concluded that the inclusion of N. limnetica enriches the egg yolk without negatively affecting the performance of laying hens and egg characteristics. Due to the positive effect on feed intake, microalgae in the diet provide nutritional benefits for laying hens. However, the positive effects of microalgae, especially on the health of laying hens, warrants further research.

Dietary protein sources can have profound effects on host-microbe interactions in the gut that are critically important for immune resilience. However more knowledge is needed to assess the impact of different protein sources on gut and animal health. Thirty-six wildtype male C57BL/6J mice of 35 d age (n = 6/group; mean ± SEM body weight 21.9 ± 0.25 g) were randomly assigned to groups fed for four weeks with semi synthetic diets prepared with one of the following protein sources containing (300 g/kg as fed basis): soybean meal (SBM), casein, partially delactosed whey powder, spray dried plasma protein, wheat gluten meal and yellow meal worm. At the end of the experiment, mice were sacrificed to collect ileal tissue to acquire gene expression data, and mammalian (mechanistic) target of rapamycin (mTOR) activity, ileal digesta to study changes in microbiota and serum to measure cytokines and chemokines. By genome-wide transcriptome analysis, we identified fourteen high level regulatory genes that are strongly affected in SBM-fed mice compared to the other experimental groups. They mostly related to the mTOR pathway. In addition, an increased (P < 0.05) concentration of granulocyte colony-stimulating factor was observed in serum of SBM-fed mice compared to other dietary groups. Moreover, by 16S rRNA sequencing, we observed that SBM-fed mice had higher (P < 0.05) abundances of Bacteroidales family S24-7, compared to the other dietary groups. We showed that measurements of genome-wide expression and microbiota composition in the mouse ileum reveal divergent responses to diets containing different protein sources, in particular for a diet based on SBM.

An experiment was performed to study the effects of a low inclusion level of Chlorella vulgaris (CV) biomass in broiler diets on performance, immune response related to inflammatory status, and the intestinal histomorphology. The study was performed with 120 Ross 308 male broiler chickens from 0–35 days of age. The broilers were housed in 12 floor pens (1.5 m²) bedded with wood shavings. The broilers received a three phase diet program, either with 0.8% CV biomass (CV) or without CV (CON). Each diet program was replicated in six pens. The final body weight increased (p = 0.053), and the feed conversion ratio (FCR), corrected for body weight, was reduced (p = 0.02) in birds fed CV compared to birds fed CON. In addition, decreased haptoglobins (p = 0.02) and interleukin-13 (p < 0.01) responses were observed during the grower phase of birds fed CV compared to the birds fed CON. A strong correlation (r = 0.82, p < 0.01) was observed between haptoglobin response and FCR. Histomorphology parameters of the jejunum were not different between the groups. It was concluded that the inclusion of 0.8% CV biomass in broiler diets is effective in influencing immune responses related to inflammatory status and promoting broiler growth.

Replacement of current protein resources for novel or alternative sources may be one of the solutions to abolish the expected scarcity of dietary protein for animal feeds. However, little is known about the nutritional, protein composition, and potential functional value of such novel or alternative protein sources. In the present study, we used advanced proteomic and bioinformatic approaches to characterize the protein component of 6 different protein sources and predicted the biofunctionalities of their in vivo digestion products. We used casein (CAS), partially delactosed whey powder (DWP), spray-dried porcine plasma (SDPP), soybean meal (SBM), wheat gluten meal (WGM), and yellow meal worm (YMW). We characterized and semiquantified the individual proteins present in these resources by nano-liquid chromatography–LTQ Orbitrap–mass spectrometry (nLCMS). Based on the data obtained, we calculated the AA composition of the proteins that constitute 90% of the total calculated protein content and compared this with the chemically determined AA compositions of the corresponding protein sources. By the use of bioinformatic procedures, we predicted the bioactive properties of these protein sources after in silico digestion with monogastric proteolytic enzymes. We detected and semiquantified 37, 58, 85, 188, 113, and 33 different individual proteins in CAS, DWP, SDPP, SBM, WGM, and YMW, respectively. The calculated AA composition of the various protein sources was almost identical to the chemically determined composition, with correlation values ranging from 0.85 to 0.94. Furthermore, we revealed that the selected protein sources are potentially rich in bioactive peptides, in particular of angiotensin-converting enzyme inhibitors and peptides with antioxidative properties. We discuss the results in terms of the benefit of the applied nLCMS-based approach for analyzing protein feed ingredients and the use of these alternative sources of protein in animal feeds for monogastrics. Furthermore, we discuss new potential applications of this method in the area of (animal) nutrition. Copyright © 2016. American Society of Animal Science .