Effects of Dietary Supplementation with Mulberry (Morus alba L.) Leaf Polysaccharides on Immune Parameters of Weanling Pigs.

In this study, the effect of dietary supplementation of mulberry leaf polysaccharides (MLPs) on the immune parameters-i.e., the immune organ weight, serum immunoglobulins, cytokines, nitric oxide (NO) production, and insulin-Like growth factor-1 (IGF1) mRNA expression-of weanling pigs as a model animal was investigated. A total of 120 healthy weanling pigs (aged 28 ± 2 d) with the same body weights were randomly divided into four groups: (1) Control treatment (CT), basal diet (BD), (2) MLP low-dose treatment (MLT), 0.6 g/kg MLP + BD, (3) MLP high-dose treatment (MHT), 1.2 g/kg MLP + BD, and (4) antibiotic treatment (AT), 0.15 g/kg chlortetracycline + BD. The results revealed that the thymus and spleen indices were significantly increased (P < 0.05) in both MLT and MHT groups in comparison with the CT group, while the serum levels of immunoglobulin G (IgG), interleukin (IL)-1ß, IL-2, IL-8, and interferon (IFN-γ) in the MLT group and IL-2, IL-6, and IFN-γ in the MHT group were also considerably greater (P < 0.05) than the corresponding levels in the CT group. The serum contents of IgG, IL-1ß, IL-2, and IL-8 in the MLT group and IL-2 and IL-6 in the MHT group were significantly increased in comparison with the corresponding contents in the AT group (P < 0.05). The transformation rate of lymphocytes in the MLT and MHT groups was higher compared to the CT and AT groups. However, a notable difference was found between the MLT group and the two control groups. The peripheral lymphocyte NO production in the MLT, MHT, and AT groups was significant relative to the CT group. The expression levels of IGF1 mRNA in the liver and muscle longissimus tissues of both the MLT and MHT groups showed significant improvement (P < 0.05) over those in the CT group. Moreover, the IGF1 mRNA expression in the muscle longissimus from the MLT group was significantly higher than in the AT group. In conclusion, the results suggest that incorporating MLPs into the diets of weanling pigs improves the animals' metabolisms and immune functions, and the effects of the MLT group were superior to those of both the MHT and AT groups.

Selectivity of deproteinization and demineralization using natural deep eutectic solvents for production of insect chitin (Hermetia illucens).

Selectivity of deproteinization and demineralization using natural deep eutectic solvents for preparation of insect chitin (Hermetia illucens) was investigated. The relationships between the pH and pKa values of NADESs and demineralization, deproteinization and crystallinity indexes of the obtained chitin were discussed, respectively. It was found that the acidic NADESs consist of basic hydrogen bond acceptor (HBA) and acidic hydrogen bond donor (HBD), or the alkali NADESs composed of acid HBA and alkali HBD were better for chitin products. After reused for three times, demineralization and deproteinization abilities of NADESs were not significantly decreased. Mechanistic analysis indicated that H+ released from HBA or HBD was the main reason for demineralization, whereas intermolecular and intramolecular hydrogen bond in NADESs facilitates the removal of protein. Overall, this report provides a preliminary reference for design of the sustainable NADESs for preparation of chitin from natural resources.

Identification and validation of reference genes for qRT-PCR analysis in mulberry (Morus alba L.).

Mulberry (Morus alba L.) is an important economic tree species in many countries. Quantitative real time PCR (qRT-PCR) has become a widely used method for gene expression studies in plants. A suitable reference gene is essential to ensure accurate and reliable results for qRT-PCR analyses. However, no reports describing the selection of reference genes have been published for mulberry. In this work, we evaluated the stability of twenty candidate reference genes in different plant tissues and under different stress conditions by qRT-PCR in mulberry using algorithms in two programs-geNorm and NormFinder. The results revealed that TUB2, UBI4, ACTIN3 and RPL4 were ranked as the most stable reference genes in the samples subsets, whereas EF1α4 and TUB3showed the least stability with both algorithms. To further validate the stability of the reference genes, the expression patterns of six genes of mulberry were analyzed by normalization with the selected reference genes. Our study will benefit future analyses of gene expression in mulberry.

Proteomic and immunological identification of two new allergens from silkworm (Bombyx mori L.) pupae.

This study explored food allergy caused by eating silkworm (Bombyx mori L.) pupae, a traditionally accepted food and animal feed in East and Southeast Asia, and identified two new allergens by proteomic and immunological methods. Proteins isolated from silkworm pupae were separated by two-dimensional gel electrophoresis (2-DE); pooled sera from patients allergic to silkworm pupa proteins were used to detect immunoglobulin E (IgE)-binding proteins by western blotting, and allergens specific for silkworm pupa consumption-caused allergy were visualised with the ECL reagents. The selected allergen proteins were further identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. Finally, chitinase and paramyosin were identified as silkworm pupa proteins showing strong immunoglobulin (IgE)-binding reaction. Analysis of the sequence homology of the two proteins using the AllergenOnline database indicated that chitinase and paramyosin shared 24.8% and 62.8% sequence homology with known allergens Der f 18 (Dermatophagoides farinae) and Der p 11 (Dermatophagoides pteronyssinus), respectively. Our results shed light on the understanding and treatment of silkworm pupa allergy.

CpG oligodeoxynucleotide protect neonatal piglets from challenge with the enterotoxigenic E. coli.

CpG motifs activates mammalian lymphocytes and macrophages to produce cytokines and polyclonal Ig. These include IFN-γ, IL-12, TNF-a, which are important in the control of bacterial infection. But thus far, the innate immunostimulatory effects of CpG ODN against pathogen have been established mainly in mouse, monkey, sheep, chicken, but not in neonatal piglets. The purpose of this study is to determine the potential protection of CpG ODN against enterotoxigenic Escherichia coli (ETEC) (with which neonatal piglets were susceptible to infection in our lab) in neonatal piglets. Here, we show intranasal (IN)-mucosal and intramuscularly (IM) systemic administration of CpG ODN could enhance innate cellular (cytokine) immunity in the sera and intestine mucosa post challenge, and thereafter the development of antigen-specific antibodies in piglets. IN and IM immunizations of neonatal piglets without antigen both reduced the ETEC excretion and alleviated diarrhoea symptoms upon challenge, and IN route had better protection effects than IM route. Protection in this study was linked to induction of a Th1 response which induced by CpG ODN. Co-delivery with Emulsigen (EM), could improve protection mediated by CpG ODN. These observations indicate that IN administration of 100 µg/kg CpG ODN with 20% EM codelivery may represent a valuable strategy for induction of innate immunity against ETEC infection in neonatal piglets.