饲粮α-亚麻酸水平对意大利蜜蜂工蜂幼虫生理机能的影响

doi:10.3864/j.issn.0578-1752.2019.13.015

Abstract

Abstract:

【Objective】 The objective of this study is to explore the effect of dietary α-linolenic acid levels on the antioxidant activity and immunity of Apis mellifera ligustica worker bee larvae. 【Method】 Twelve hundreds 1-day-old worker bee larvae were randomly divided into 5 group, with 5 replicates in each group and 48 worker bee larvae in each replicate. The control group was fed with basic diets without α-linolenic acid, and the 4 treatment groups were fed experimental diets containing different amounts of α-linolenic acid (0.02%, 0.04%, 0.06%, and 0.08%). According to the indoor feeding in vitro method, the 1-day-old worker bee larvae were moved to a 24-well cell culture plate with 200 μL diet at suitable temperature. The larvae were moved by worm-moving needle. The culture plate was placed in a constant temperature incubator (temperature 33℃, relative humidity 55%), the diet was changed daily during the test. Feeding until the end of the 6th day or the beginning of the 7th day, when the larvae started to stand upright or defecate, the larvae were moved to the 24-well cell culture plate covered with sterilized paper in advance to prepare for pupation. From the 1st day of feeding, the death number of larvae and pupae was checked and recorded every day, and the dead individuals were removed in time until all adult emerged. The number of undead pupae and emergence new bees was recorded, and pupation rate and emergence rate were calculated. The 5-, 6- and 7-day-old worker bee larvae were used to measure antioxidant, immune, lipid metabolism indexes and related gene expression. 【Result】 When the addition level of α-linolenic acid in the diet was 0.02% and 0.04%, the pupation rate and emergence rate were significantly higher than those in other treatment groups (P＜0.05), while the contents of triglyceride (TG), total cholesterol (TC) and low density lipoprotein (LDL) in the hemolymph of worker bee larvae were significantly lower than those of the control group, the content of high density lipoprotein (HDL) was significantly higher than that of the control group (P＜0.05). Compared with the control group, the activity of superoxide dismutase (SOD) was significantly increased, while the content of malondialdehyde (MDA) was significantly decreased when the addition level of α-linolenic acid was 0.04% (P＜0.05). When the addition level of α-linolenic acid was 0.02%, 0.04% and 0.06%, the activities of lysozyme and phenoloxidase (PO) of 6-day-old worker bee larvae were significantly higher than those in the control group (P＜0.05). When the addition level of α-linolenic acid was 0.04%, the activities of fatty acid synthase (FAS) and acetyl CoA carboxylase (ACC) of 6-day-old worker bee larvae were significantly lower than those in the control group (P＜0.05). When the addition level of α-linolenic acid was 0.04%, the relative expression levels of lysozyme and PO of 5- and 7-day-old worker bee larvae were significantly higher than those in the control group, but when the addition level of α-linolenic acid was 0.08%, the relative expression level of lysozyme was significantly decreased (P＜0.05). 【Conclusion】 Different dietary supplementation levels of α-linolenic acid have significant effects on antioxidant activity and immunity of A. m. ligustica worker bee larvae, and the appropriate addition level of α-linolenic acid in larval diet is 0.02%-0.04%.

Key words: Apis mellifera ligustica, worker bee, α-linolenic acid, addition level, antioxidant activity, immunity

YU Jing,ZHANG WeiXing,MA LanTing,XU BaoHua. Effect of Dietary α-Linolenic Acid Levels on Physiological Function of Apis mellifera ligustica Worker Bee Larvae[J].Scientia Agricultura Sinica, 2019, 52(13): 2368-2378.

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原料 Ingredient	α-亚麻酸添加水平 Level of α-linolenic acid (%)
原料 Ingredient	0	0.02	0.04	0.06	0.08
蜂王浆 Royal jelly 葡萄糖 Glucose 果糖 Fructose 酵母提取物 Yeast extract 无菌水 Sterile water α-亚麻酸α-linolenic acid 合计Total	50.00 6.00 6.00 1.00 37.00 0 100.00	50.00 6.00 6.00 1.00 37.00 0.02 100.00	50.00 6.00 6.00 1.00 37.00 0.04 100.00	50.00 6.00 6.00 1.00 37.00 0.06 100.00	50.00 6.00 6.00 1.00 37.00 0.08 100.00

目的基因 Target gene	引物序列 Primer sequence (5′-3′)	产物长度 Product length (bp)	GenBank登录号 GenBank accession number
酚氧化酶基因 PO	F: ATGAGGACTTCCTGTCTGTTGCAG R: CTTCGGTAAGCGGTGGTACTGG	132	XM_026444741.1
溶菌酶基因 lysozyme	F: TCGCACTACTCGCCACGATA R: TTTGATGCAGAGCGAGAGCG	114	NM_001011614.1

项目 Item	α-亚麻酸添加水平Level of α-linolenic acid (%)					P值 P-value
项目 Item	0	0.02	0.04	0.06	0.08	P值 P-value
化蛹率 Pupation rate (%)	82.86±0.80a	86.31±1.07a	84.65±0.26a	73.70±0.71b	51.14±0.56c	<0.0001
羽化率 Emergence rate (%)	60.00±0.39a	62.45±0.86a	52.50±0.35b	23.33±1.33c	16.67±0.66d	<0.0001

项目 Item	α-亚麻酸添加水平Level of α-linolenic acid (%)					P值 P-value
项目 Item	0	0.02	0.04	0.06	0.08	P值 P-value
总蛋白TP (g·L^-1)	40.80±0.37ab	39.38±1.31b	36.08±1.27c	42.42±0.74a	40.70±1.36ab	0.0013
甘油三酯TG (mmol·L^-1)	3.36±0.39a	1.59±0.04b	1.58±0.01b	2.01±0.21b	1.56±0.13b	0.0002
总胆固醇TC (mmol·L^-1)	0.94±0.01a	0.59±0.03c	0.63±0.02c	0.77±0.01b	0.78±0.03b	<0.0001
高密度脂蛋白HDL (mmol·L^-1)	0.38±0.01c	0.60±0.01b	0.79±0.03a	0.66±0.07b	0.40±0.03c	<0.0001
低密度脂蛋白LDL (mmol·L^-1)	0.17±0.01a	0.13±0.01b	0.09±0.01c	0.14±0.01b	0.18±0.00a	<0.0001

项目 Item	α-亚麻酸添加水平 Level of α-linolenic acid (%)					P值 P-value
项目 Item	0	0.02	0.04	0.06	0.08	P值 P-value
5日龄幼虫 5-day-old larva	7.89±0.54c	13.80±0.47b	9.50±0.40c	16.30±0.23a	13.62±1.47b	0.0002
7日龄幼虫 7-day-old larva	12.05±0.82c	13.49±0.11abc	12.35±0.07bc	14.61±0.87a	15.76±1.01a	0.0190

Effect of Dietary α-Linolenic Acid Levels on Physiological Function of Apis mellifera ligustica Worker Bee Larvae

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