Nutrition Profile and Animal-Tested Safety of Morchella esculenta Mycelia Produced by Fermentation in Bioreactors.

Morchella esculenta (ME), or "true" morel mushrooms, are one of the most expensive mushrooms. M. esculenta contain all the important nutrients including carbohydrates, proteins, polyunsaturated fatty acids, and several bioactive compounds such as polysaccharides, organic acids, polyphenolic compounds, and tocopherols, which are promising for antioxidant, immunomodulation, anti-cancer, and anti-inflammatory applications. However, the M. esculenta fruiting body is difficult to collect in nature and the quality is not always reliable. For this reason, the cultivation of its mycelia represents a useful alternative for large-scale production. However, for M. esculenta mycelia to be used as an innovative food ingredient, it is very important to prove it is safe for human consumption while providing high-quality nutrients. Hence, for the first time in this study, the nutritional composition, as well as 90 days of oral toxicity of fermented ME mycelia in Sprague Dawley rats, is examined. Results showed that the ME mycelia contained 4.20 ± 0.49% moisture, 0.32 ± 0.07% total ash, 17.17 ± 0.07% crude lipid, 39.35 ± 0.35% crude protein, 38.96 ± 4.60% carbohydrates, and 467.77 ± 0.21 kcal/100 g energy, which provides similar proportions of macronutrients as the U.S. Dietary Reference Intakes recommend. Moreover, forty male and female Sprague Dawley rats administrating ME mycelia at oral doses of 0, 1000, 2000, and 3000 mg/kg for 90 days showed no significant changes in mortality, clinical signs, body weight, ophthalmology, and urinalysis. Although there were alterations in hematological and biochemical parameters, organ weights, necropsy findings, and histological markers, they were not considered to be toxicologically significant. Hence, the results suggest that the no-observed-adverse-effects level (NOAEL) of ME mycelia was greater than 3000 mg/kg/day and can therefore be used safely as a novel food at the NOAEL.

Effects of Deinococcus spp. supplement on egg quality traits in laying hens.

To counter the ill effects of synthetic dyes, bacterial pigment production as an alternative is now one of the promising and emerging fields of research. This study was conducted to evaluate the applicability of Deinococcus genus on the egg quality traits in laying hens. In study I, 24 single comb White Leghorn layers were fed with various 1 wt % Deinococcus bacterial strains for 10 d. In study II, 84 brown Hendrix layers were fed with one of 4 diets containing 0, 0.2, 1, or 5 wt % Deinococcus sp. GKB-Aid 1995 powder for 12 wk. In study III, 60 White Leghorn laying hens were fed either with or without 1 wt % Deinococcus sp. GKB-Aid 1995 powder, 1 wt % Deinococcus sp. GKB-Aid 1995 granules, or 1 wt % Deinococcus sp. GKB-Aid 1995 oily granules for 10 successive d. In all of the experiments, feeding Deinococcus powder did not affect egg quality traits except for the yolk color. In particular, supplementation with all Deinococcus powder treatments changed the yolk color (P < 0.05) in study I, with the best pigmentation score obtained by D. grandis and Deinococcus sp. GKB-Aid 1995. Moreover, longer supplementation of Deinococcus sp. GKB-Aid 1995 in study II had a significant effect on feed conversion ratio. With these findings under consideration, the present study suggests that the Deinococcus species, especially Deinococcus sp. GKB-Aid 1995, can be an excellent candidate for improving egg yolk color in laying hens.

Characterization and safety evaluation of a Deinococcus member as feed additive for hens.

As previous studies mainly focus on understanding the mechanisms of radioresistance in Deinococcus bacteria, the present study aimed at characterizing and verifying the safety use of the GKB-Aid 1995 strain, a member of the radiation-resistant bacterial genus Deinococcus, as an ingredient in feed supplements. Using Vitek 2 system and 16S rRNA gene sequencing, GKB-Aid 1995 most resembles Deinococcus grandis. The Ames test, in vitro chromosomal test, in vivo micronucleus test and acute toxicity test were performed subsequently for its safety evaluation. As there is a possibility that the pigment of GKB-Aid 1995 can pass from feed to eggs intended for human consumption, an acute toxicity test was also carried out in pigmented egg yolk. The results confirmed that GKB-Aid 1995 was non-genotoxic in three genotoxicity experiments, and the LD50 of GKB-Aid 1995 and the pigmented egg yolk in ICR mice was greater than 10 and 12 g kg(-1) body weight, respectively. Overall, these data indicate that GKB-Aid 1995 is a non-toxic substance with no genotoxicity and is therefore safe to be used as a feed supplement or feed additive. This study suggests there is potential in developing GKB-Aid 1995 as an animal feed additive intended to enhance yolk coloration to meet the demand of consumers.

The expression of two novel orange-spotted grouper (Epinephelus coioides) TNF genes in peripheral blood leukocytes, various organs, and fish larvae.

The tumour necrosis factor (TNF) super-family is a group of important cytokines involved in inflammation, apoptosis, cell proliferation, and the general stimulation of the immune system. The TNF gene has been cloned in some bony fish; however, its counterparts are still unidentified in the majority of fish species. In this study, we cloned gTNF-1 and gTNF-2 from the orange-spotted grouper (Epinephelus coioides), an economically important farmed fish. Both genes include 4 exons and 3 introns and encoded 253 and 241 amino acid proteins with a molecular weight of approximately 27 and 26 kDa, respectively. The identity of the putative amino acid sequences between gTNF-1 and gTNF-2 was only 38%. The positions of cysteine residues, a protease cleavage site, and a transmembrane domain sequence derived from gTNF-1 and gTNF-2 were similar to those in other fish and mammalian TNF-α. The mRNA expression levels of the 2 gTNF molecules were evaluated in unstimulated/stimulated peripheral blood leukocytes, various organs, and fish larvae. Following lipopolysaccharide (LPS) treatment, gTNF-2 was expressed at higher levels, was up-regulated more quickly, and was more sensitive to the immune response than gTNF-1. gTNF-1 was constitutively expressed in the thymus, brain, and spleen, but it was also expressed in the heart, head kidney, and trunk kidney after LPS stimulation. gTNF-2 was constitutively expressed in the thymus, head kidney, trunk kidney, spleen, and intestine; further, gTNF-2 was highly expressed in all organs post-LPS stimulation. Finally, the gTNF expression levels were evaluated at various developmental stages in grouper larvae. A higher variation of gTNF expression levels was observed in fish larvae from a contaminated hatchery. This study revealed the different expression patterns of gTNF-1 and gTNF-2. In addition, gTNF-2 was more sensitive to pathogens than gTNF-1; therefore, it may be an appropriate marker for pathogen invasion and the evaluation of the larval rearing environment.