锰对意大利蜜蜂成年工蜂生理机能的影响

Effects of Manganese on Physiological Function of Adult Worker Bees for Apis mellifera L.

本试验以意大利蜜蜂(Apis mellifera L.)为研究对象,通过在成年工蜂饲粮中添加不同水平的锰,开展室内离群成年工蜂饲养试验,对寿命、机体生理生化指标以及锰相关基因表达量进行测定,力求探明锰对蜜蜂的营养作用及机制。从意大利蜜蜂姊妹蜂群中取1250只刚羽化出房的1日龄工蜂,随机分为5组,每组5个重复,每个重复50只。对照组蜜蜂供给不额外添加锰的糖浆饲粮;4个试验组蜜蜂供给添加不同水平葡萄糖酸锰的糖浆饲粮,使饲粮中锰添加水平分别为50、100、150、200 mg/kg。将各组蜜蜂放入专门的蜜蜂饲养装置中,每个饲养装置放入50只蜜蜂,置于恒温恒湿培养箱,设置培养箱内温度为30℃,相对湿度为(50±5)%,每天定时用注射器饲喂对应的糖浆饲粮3 mL,分别在蜜蜂7、14、21日龄时取样,测定成年工蜂体内锰相关酶活性及锰相关基因表达量;另在蜜蜂21日龄时取样,测定成年工蜂体内锰含量。另从意大利蜜蜂姊妹蜂群中取1250只刚羽化出房的1日龄工蜂,按照上述处理进行分组和饲喂,观察蜜蜂表现,清除死亡蜂体,饲喂至全部死亡,统计各组蜜蜂的平均寿命。结果显示:1)50、100 mg/kg锰添加组的平均寿命显著长于对照组及150、200 mg/kg锰添加组(P<0.05)。2)随着饲粮中锰添加水平的升高,21日龄成年工蜂体内锰含量显著增加(P<0.05)。3)随着饲粮中锰添加水平的升高,7、14日龄成年工蜂体内锰超氧化物歧化酶(MnSOD)活性均呈现先升高后下降的趋势,并在锰添加水平为150 mg/kg时MnSOD活性达到最高且显著高于对照组(P<0.05);7、14日龄成年工蜂体内锰过氧化氢酶(MnCAT)活性均呈现先升高后下降的趋势,21日龄成年工蜂体内MnCAT活性则呈上升趋势,且在锰添加水平为100 mg/kg时14日龄成年工蜂体内MnCAT活性达到最高且显著高于对照组(P<0.05);7、14、21日龄成年工蜂体内精氨酸酶活性均呈现先升高后下降的趋势。4)随着饲粮中锰添加水平的升高,7、14、21日龄成年工蜂体内MnSOD基因的表达量均呈现先上调后下调的趋势,且均在锰添加水平为150 mg/kg时达到最高并在7和21日龄时显著高于其他各组(P<0.05);14和21日龄成年工蜂体内MnCAT基因的表达量呈现先上调后下调的趋势,并在锰添加水平为100 mg/kg时显著高于对照组(P<0.05)。此外,14日龄时,150 mg/kg锰添加组成年工蜂体内精氨酸酶基因的表达量显著高于对照组和200 mg/kg锰添加组(P<0.05);21日龄时,200 mg/kg锰添加组成年工蜂体内精氨酸酶基因的表达量显著高于对照组和50、100 mg/kg锰添加组(P<0.05)。由此得出,饲粮中补充适量的锰能延长意大利蜜蜂成年工蜂的寿命,提高机体抗氧化能力、免疫能力等生理机能。

This experiment used Apis mellifera L.as a research object to carry out indoor out-of-group adult worker bee feeding experiments by adding different levels of manganese to the adult worker bee diet.The lifespan,physiological and biochemical indexes and manganese related gene expression of the bees were measured,and strive to find the nutritional effects and mechanisms of manganese on bees.From the Apis mellifera L.sister bee colony,a total of 1250 one-day-old work bees that had just emerged from the house were randomly divided into 5 groups with 5 replicates in each group and 50 bees per replicate.One group was control group,and bees in the control group were fed a syrup diet without manganese.The other four groups were experimental groups,and bees in the experimental group were fed the syrup diet supplemented with different levels of manganese gluconate,and the levels of manganese in the syrup diets were 50,100,150 and 200 mg/kg,respectively.Put 50 bees in each group into a special bee feeding device and placed them in a constant temperature and humidity incubator.Set the temperature in the incubator to 30℃ and the humidity of(50±5)%.Regularly used a syringe to fed 3 mL of syrup diet,each group was sampled at 7,14 and 21 days of age,and the manganese-related enzyme activities and manganese-related gene expression levels in adult worker bee body were measured.Moreover,samples were taken at 21 days of age to determine the manganese content in adult worker bee body.From the Apis mellifera L.sister bee colony,another 1250 one-day-old work bees that had just emerged from the house were grouped and fed according to the above treatment,observed bee performance,removed dead bee body,and fed until all death.The average lifespan of each group of bees was counted.The results of showed as follows:1)the average lifespan in the 50 and 100 mg/kg manganese addition groups was significantly longer than that in the 150 and 200 mg/kg manganese addition groups and the control group(P<0.05).2)With the supplemental level of manganese in the diet increasing,the manganese content in adult worker bee body was significantly increased(P<0.05).3)With the supplemental level of manganese in the diet increasing,the activity of manganese superoxide dismutase(MnSOD)in the 7-and 14-day-old adult worker bee body was firstly increased and then decreased.When the supplemental level of manganese was 150 mg/kg,the activity of MnSOD reached the highest value and significantly higher than that in the control group(P<0.05).The manganese catalase(MnCAT)activity in the 7-and 14-day-old adult worker bee body was firstly increased and then decreased.And the MnCAT activity in the 21-day-old adult worker bee body was increased.When the supplemental level of manganese was 150 mg/kg,the activity of MnCAT in the 14-day-old adult worker bee body reached the highest value and was significantly higher than that in the control group(P<0.05).The activity of arginase in the adult worker bee body at 7,14 and 21 days of age was firstly increased and then decreased.4)With the supplemental level of manganese in the diet increasing,the expression level of MnSOD gene in the adult worker bee body at 7,14 and 21 days of age was firstly increased and then decreased.When the manganese supplemental level was 150 mg/kg,the expression level of MnSOD gene in the adult worker bee body at 7 and 14 days of age was significantly higher than that in the other groups(P<0.05).The expression levels of MnCAT gene in 14-and 21-day-old adult worker bee body was firstly increased and then decreased.When the manganese supplemental level was 100 mg/kg,the expression level of MnCAT gene in 14-and 21-day-old adult worker bee body was significantly higher than that in the control group(P<0.05).In addition,the expression level of arginase gene in 14-day-old adult worker bee body in 150 mg/kg manganese addition group was significantly higher than that in the control group and 200 mg/kg manganese addition group(P<0.05),and the expression level of arginase gene in 21-day-old adult worker bee body in 200 mg/kg manganese addition group was significantly higher than that in the control group and 50,150 mg/kg manganese addition groups(P<0.05).The results indicate that supplementing an appropriate amount of manganese in the diet can prolong the lifespan,improve the antioxidant capacity,immunity and other physiological functions of adult worker bees for Apis mellifera L.