饲喂漂白粉液浸消叶对蚕体生理及肠道产酶菌的影响

发布时间：2018-05-29 19:04

本文选题：漂白粉 + 家蚕　；参考：《西南大学》2017年硕士论文

【摘要】：用漂白粉液浸消桑叶全程饲喂家蚕已经成为有效防控微粒子病的方法,被广泛应用在蚕种生产上。但家蚕饲喂消毒桑叶后出现了体质减弱、饲育成绩下降等现象,针对这些问题,学者们从家蚕饲育成绩方面着手进行了探究,但更深层次的关于家蚕生理方面及肠道微生物菌群变化的研究鲜有报道。本实验用不同有效氯浓度(0.3%、0.4%、0.5%、0.6%)漂白粉液浸消后晾干的桑叶饲喂4龄起蚕直到其上蔟,以清水组为对照,另设全程0.3%处理组,即从收蚁至上蔟全程饲喂0.3%有效氯浓度漂白粉液浸消叶。调查统计了家蚕饲喂浸消叶后幼虫期生长发育指标、茧期相关生理指标的变化;测定了5龄1天、3天、5天、7天家蚕肠液蛋白酶、淀粉酶、脂肪酶的活性及血液超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)的活性,并比较分析不同处理组与对照组之间的差异;用NA培养基分离纯化了家蚕肠道细菌,分析了不同处理组与对照组家蚕肠道细菌数量的差异;利用不同筛选培养基研究了不同处理组与对照组家蚕肠道产消化酶(蛋白酶、淀粉酶、脂肪酶、纤维素酶)细菌的菌株数量、产酶种类及相对产酶能力等的差异;用16S rDNA测序技术对产酶菌分离株进行了鉴定,比较不同处理组及对照组产酶菌种类的不同。通过实验,主要得到以下结果:1、从4龄起饲喂漂白粉液浸消叶的实验组中,有效氯浓度为0.3%时明显促进了4龄家蚕体重增长;有效氯浓度为0.5%和0.6%时显著抑制了4龄家蚕体重的增长,显著减弱了5龄家蚕消化量和消化率;而全程饲喂0.3%有效氯浓度漂白粉液浸消叶时显著降低了5龄家蚕的消化量和消化率;各处理组家蚕结茧率均显著低于对照,死笼率均显著高于对照,其中全程0.3%组对结茧率影响最大。以上说明,目前蚕种生产上采用的全程饲喂0.3%有效氯浓度漂白粉液浸消叶的消毒方式会影响家蚕的消化吸收,进而影响蚕体健康,对家蚕生理功能有一定影响。2、饲喂不同有效氯浓度漂白粉液浸消叶对家蚕肠液消化酶和血液保护酶活性的影响实验结果表明,有效氯浓度为0.3%时家蚕肠液蛋白酶和淀粉酶活性受到抑制,尤其淀粉酶活性受到显著抑制,血液SOD、CAT、POD活性均显著上升;0.4%、0.5%、0.6%有效氯浓度下,血液POD、CAT活性均显著上升,家蚕肠液蛋白酶和淀粉酶活性显著下降,脂肪酶活性显著升高,推测是家蚕抵御有效氯胁迫的一种抗性反应。而全程饲喂0.3%有效氯浓度漂白粉液浸消叶后家蚕肠液蛋白酶和淀粉酶活性受到抑制,尤其淀粉酶活性受到显著抑制,血液CAT和POD活性显著上升。说明全程饲喂0.3%有效氯浓度漂白粉液浸消叶后影响了家蚕对桑叶蛋白和糖类的消化吸收,尤其对淀粉的消化受到显著抑制。此外,漂白粉的强氧化性对蚕体构成一种胁迫,使家蚕血液保护酶体系活性增强,相互协调,保护蚕体免受自由基损伤。3、饲喂各有效氯浓度漂白粉液浸消叶均显著抑制了家蚕肠道细菌菌落数量,且随有效氯浓度升高抑制增强,全程0.3%组影响显著。饲喂漂白粉液浸消叶后,家蚕肠道内产蛋白酶和淀粉酶菌株数量减少,相对产酶能力减弱,产脂肪酶菌株只在0.6%组和全程0.3%组中筛选到,且0.6%组产脂肪酶菌株产酶能力明显高于全程0.3%组,推测脂肪酶可能与蚕体抵抗有效氯胁迫的抗性有关。饲喂浸消叶后家蚕肠道产消化酶细菌类群构成有一定变化,清水组和全程0.3%组产酶菌属有葡萄球菌属Staphylococcus sp.、Lelliottia sp.、布丘氏菌属Buttiauxella sp.;其他处理组的产酶菌属均只有葡萄球菌属Staphylococcus sp.,以上说明有效氯胁迫抑制了产蛋白酶和淀粉酶菌株的生长,产酶菌类群单一,这可能和肠道内消化酶活性的变化及家蚕的营养消化吸收相关,而全程0.3%组由于长期饲喂消毒桑叶导致蚕体已经形成对有效氯胁迫的应对方式。4、生产上用0.3%有效氯浓度漂白粉液全程浸消桑叶的消毒方式致使家蚕肠液蛋白酶、淀粉酶活性下降,抑制了肠道产蛋白酶和淀粉酶菌株的数量和产酶能力,影响了家蚕对营养物质的消化吸收,蚕体血液抗氧化性保护酶POD、CAT活性在有效氯胁迫下升高,总体来看,家蚕的体质下降、结茧率下降、死笼率升高,影响了家蚕的生长发育和正常生理活动。因此,蚕种生产上应该改进漂白粉的使用方式,或筛选更有效的蚕用药物,以保证蚕种生产的安全进行。
[Abstract]:The whole course of feeding silkworm in mulberry leaves with bleaching powder has become an effective method to prevent and control the disease of microparticle, and it has been widely used in the production of silkworm. However, the silkworm was fed by the silkworm to disinfect the mulberry leaves. There are few reports about the changes in the physiological and intestinal microflora of the silkworm. In this experiment, the dried mulberry leaves were fed with different effective chlorine concentrations (0.3%, 0.4%, 0.5%, 0.6%) to feed 4 age silkworms until they were used as control, and the whole course 0.3% treatment group was set up in the whole course, that is to feed 0.3% effective chlorine concentration from the ant to the top. The growth and development indices of the silkworm larvae and the changes of related physiological indexes in the cocoon period were investigated, and the activity of protease, amylase, lipase, SOD, CAT and POD activity of silkworm in the silkworm, 5 instar, 1 days, 3 days, 5 days and 7 days were investigated and analyzed. The differences between the different treatment groups and the control group were compared and analyzed. The intestinal bacteria in the silkworm of the silkworm were isolated and purified by NA medium, and the difference of intestinal bacteria in the silkworm was analyzed between the different treatment groups and the control group, and the digestive enzymes (protease, amylase, lipase, fiber, fiber, fiber) of the silkworm in the different treatment groups and the control group were studied by different screening medium. The difference in the number of bacteria strains, the species of enzyme producing and the ability of relative enzyme production, and 16S rDNA sequencing technology was used to identify the isolated strains of enzyme producing strains, and the difference between the different treatment groups and the control group was compared. Through the experiment, the main results were as follows: 1, the effective chlorine concentration in the experiment group feeding the bleaching powder from 4 years old, effective chlorine concentration. The weight of silkworm, silkworm, 4 years old was significantly increased when the degree was 0.3%. When the concentration of effective chlorine was 0.5% and 0.6%, the body weight of the silkworm, silkworm, and the digestibility and digestibility of the 5 years old silkworm were significantly reduced, and the digestibility and digestibility of the 5 instar silkworms were significantly reduced when the 0.3% effective chlorine bleaching powder was fed by 0.3%. The silkworm, silkworm, 5 years old was significantly reduced. The cocoon yield was significantly lower than that of the control, and the dead cage rate was significantly higher than that of the control, among which the whole 0.3% groups had the greatest influence on the cocoon yield. The above indicated that the disinfection mode of 0.3% effective chlorine concentration bleaching powder in the whole process of silkworm breeding will affect the digestion and absorption of silkworm, and then affect the health of the silkworm body and the physiological function of silkworm. The effects of.2 on the activity of digestive enzymes and blood protective enzymes in silkworm intestinal juice were affected by different effective chlorine bleaching powder. The results showed that the activity of protease and amylase in silkworm intestinal liquid was inhibited when the effective chlorine concentration was 0.3%, especially the activity of amylase was significantly inhibited, the activity of SOD, CAT and POD in blood increased significantly, and 0.4%, 0.. 5%, 0.6% effective chlorine concentration, blood POD, CAT activity increased significantly, the silkworm intestinal fluid protease and amylase activity significantly decreased, lipase activity significantly increased, speculates that the silkworm resistance to effective chlorine stress resistance reaction. And the whole feeding of 0.3% effective chlorine concentration bleach solution after the bleaching of silkworm digestion and amylase activity of silkworm intestinal juice and amylase activity received In addition, the activity of amylase was significantly inhibited and the activity of CAT and POD in the blood increased significantly. It indicated that the digestion and absorption of silkworm to mulberry leaf protein and sugar was influenced by the 0.3% effective chlorine bleaching powder, especially the digestion of the starch, and the strong oxidation of bleaching powder was a threat to the silkworm. The activity of the silkworm blood protection enzyme system was enhanced and coordinated, and the silkworm body was protected from free radical damage.3. The number of bacterial colonies in the intestinal tract of silkworm was inhibited significantly by feeding the bleaching powder of each effective chlorine, and increased with the increase of the concentration of effective chlorine. The 0.3% groups were significantly affected by the increase of the concentration of the effective chlorine. The number of protease and amylase producing strains decreased, the relative enzyme producing ability was weakened, the lipase producing strain was only screened in 0.6% groups and 0.3% groups, and the ability of producing lipase in 0.6% groups was significantly higher than that of the whole course 0.3%. It was presumed that the lipase might be related to resistance to effective chlorine coercion in silkworm. There were certain changes in the composition of digestive enzyme bacteria. There were Staphylococcus Staphylococcus sp., Lelliottia sp., and Buttiauxella sp. in the genus Staphylococcus in clean water group and the whole process group. All the other treated groups were all only staphylococcal Staphylococcus sp., and the above indicated that effective chlorine stress inhibited proteinase and amylase production. The growth of the strain is single, which may be related to the changes of digestive enzyme activity in the intestines and the nutrient digestion and absorption of the silkworm, while the 0.3% groups of silkworms have formed a response to effective chlorine stress due to the long feeding of the mulberry leaves for a long period of time, and the 0.3% effective chlorine concentration bleaching powder is used to soak the disinfectant of mulberry leaves in the whole process. The activity of protease in silkworm intestinal fluid decreased, the amount of protease and amylase producing strain and the ability to produce enzyme were inhibited, the digestion and absorption of silkworm was influenced by the silkworm, the antioxidant enzyme POD and CAT activity in the silkworm body increased under the effective chlorine stress. In general, the constitution of the silkworm was decreased, the cocoon yield decreased and died. The increase of cage rate affects the growth and normal physiological activities of silkworm. Therefore, the use of bleaching powder should be improved and more effective silkworm drugs should be screened in sericulture production to ensure the safety of sericulture production.
【学位授予单位】：西南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S884.1

【参考文献】