本文選題：漂白粉 + 家蠶�。� 參考：《西南大學(xué)》2017年碩士論文

【摘要】：用漂白粉液浸消桑葉全程飼喂家蠶已經(jīng)成為有效防控微粒子病的方法,被廣泛應(yīng)用在蠶種生產(chǎn)上。但家蠶飼喂消毒桑葉后出現(xiàn)了體質(zhì)減弱、飼育成績(jī)下降等現(xiàn)象,針對(duì)這些問題,學(xué)者們從家蠶飼育成績(jī)方面著手進(jìn)行了探究,但更深層次的關(guān)于家蠶生理方面及腸道微生物菌群變化的研究鮮有報(bào)道。本實(shí)驗(yàn)用不同有效氯濃度(0.3%、0.4%、0.5%、0.6%)漂白粉液浸消后晾干的桑葉飼喂4齡起蠶直到其上蔟,以清水組為對(duì)照,另設(shè)全程0.3%處理組,即從收蟻至上蔟全程飼喂0.3%有效氯濃度漂白粉液浸消葉。調(diào)查統(tǒng)計(jì)了家蠶飼喂浸消葉后幼蟲期生長(zhǎng)發(fā)育指標(biāo)、繭期相關(guān)生理指標(biāo)的變化;測(cè)定了5齡1天、3天、5天、7天家蠶腸液蛋白酶、淀粉酶、脂肪酶的活性及血液超氧化物歧化酶(SOD)、過氧化氫酶(CAT)、過氧化物酶(POD)的活性,并比較分析不同處理組與對(duì)照組之間的差異;用NA培養(yǎng)基分離純化了家蠶腸道細(xì)菌,分析了不同處理組與對(duì)照組家蠶腸道細(xì)菌數(shù)量的差異;利用不同篩選培養(yǎng)基研究了不同處理組與對(duì)照組家蠶腸道產(chǎn)消化酶(蛋白酶、淀粉酶、脂肪酶、纖維素酶)細(xì)菌的菌株數(shù)量、產(chǎn)酶種類及相對(duì)產(chǎn)酶能力等的差異;用16S rDNA測(cè)序技術(shù)對(duì)產(chǎn)酶菌分離株進(jìn)行了鑒定,比較不同處理組及對(duì)照組產(chǎn)酶菌種類的不同。通過實(shí)驗(yàn),主要得到以下結(jié)果:1、從4齡起飼喂漂白粉液浸消葉的實(shí)驗(yàn)組中,有效氯濃度為0.3%時(shí)明顯促進(jìn)了4齡家蠶體重增長(zhǎng);有效氯濃度為0.5%和0.6%時(shí)顯著抑制了4齡家蠶體重的增長(zhǎng),顯著減弱了5齡家蠶消化量和消化率;而全程飼喂0.3%有效氯濃度漂白粉液浸消葉時(shí)顯著降低了5齡家蠶的消化量和消化率;各處理組家蠶結(jié)繭率均顯著低于對(duì)照,死籠率均顯著高于對(duì)照,其中全程0.3%組對(duì)結(jié)繭率影響最大。以上說明,目前蠶種生產(chǎn)上采用的全程飼喂0.3%有效氯濃度漂白粉液浸消葉的消毒方式會(huì)影響家蠶的消化吸收,進(jìn)而影響蠶體健康,對(duì)家蠶生理功能有一定影響。2、飼喂不同有效氯濃度漂白粉液浸消葉對(duì)家蠶腸液消化酶和血液保護(hù)酶活性的影響實(shí)驗(yàn)結(jié)果表明,有效氯濃度為0.3%時(shí)家蠶腸液蛋白酶和淀粉酶活性受到抑制,尤其淀粉酶活性受到顯著抑制,血液SOD、CAT、POD活性均顯著上升;0.4%、0.5%、0.6%有效氯濃度下,血液POD、CAT活性均顯著上升,家蠶腸液蛋白酶和淀粉酶活性顯著下降,脂肪酶活性顯著升高,推測(cè)是家蠶抵御有效氯脅迫的一種抗性反應(yīng)。而全程飼喂0.3%有效氯濃度漂白粉液浸消葉后家蠶腸液蛋白酶和淀粉酶活性受到抑制,尤其淀粉酶活性受到顯著抑制,血液CAT和POD活性顯著上升。說明全程飼喂0.3%有效氯濃度漂白粉液浸消葉后影響了家蠶對(duì)桑葉蛋白和糖類的消化吸收,尤其對(duì)淀粉的消化受到顯著抑制。此外,漂白粉的強(qiáng)氧化性對(duì)蠶體構(gòu)成一種脅迫,使家蠶血液保護(hù)酶體系活性增強(qiáng),相互協(xié)調(diào),保護(hù)蠶體免受自由基損傷。3、飼喂各有效氯濃度漂白粉液浸消葉均顯著抑制了家蠶腸道細(xì)菌菌落數(shù)量,且隨有效氯濃度升高抑制增強(qiáng),全程0.3%組影響顯著。飼喂漂白粉液浸消葉后,家蠶腸道內(nèi)產(chǎn)蛋白酶和淀粉酶菌株數(shù)量減少,相對(duì)產(chǎn)酶能力減弱,產(chǎn)脂肪酶菌株只在0.6%組和全程0.3%組中篩選到,且0.6%組產(chǎn)脂肪酶菌株產(chǎn)酶能力明顯高于全程0.3%組,推測(cè)脂肪酶可能與蠶體抵抗有效氯脅迫的抗性有關(guān)。飼喂浸消葉后家蠶腸道產(chǎn)消化酶細(xì)菌類群構(gòu)成有一定變化,清水組和全程0.3%組產(chǎn)酶菌屬有葡萄球菌屬Staphylococcus sp.、Lelliottia sp.、布丘氏菌屬Buttiauxella sp.;其他處理組的產(chǎn)酶菌屬均只有葡萄球菌屬Staphylococcus sp.,以上說明有效氯脅迫抑制了產(chǎn)蛋白酶和淀粉酶菌株的生長(zhǎng),產(chǎn)酶菌類群?jiǎn)我?這可能和腸道內(nèi)消化酶活性的變化及家蠶的營(yíng)養(yǎng)消化吸收相關(guān),而全程0.3%組由于長(zhǎng)期飼喂消毒桑葉導(dǎo)致蠶體已經(jīng)形成對(duì)有效氯脅迫的應(yīng)對(duì)方式。4、生產(chǎn)上用0.3%有效氯濃度漂白粉液全程浸消桑葉的消毒方式致使家蠶腸液蛋白酶、淀粉酶活性下降,抑制了腸道產(chǎn)蛋白酶和淀粉酶菌株的數(shù)量和產(chǎn)酶能力,影響了家蠶對(duì)營(yíng)養(yǎng)物質(zhì)的消化吸收,蠶體血液抗氧化性保護(hù)酶POD、CAT活性在有效氯脅迫下升高,總體來看,家蠶的體質(zhì)下降、結(jié)繭率下降、死籠率升高,影響了家蠶的生長(zhǎng)發(fā)育和正常生理活動(dòng)。因此,蠶種生產(chǎn)上應(yīng)該改進(jìn)漂白粉的使用方式,或篩選更有效的蠶用藥物,以保證蠶種生產(chǎn)的安全進(jìn)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.
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S884.1

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