汉赛巴尔通体感染长角血蜱后的体内分布及宿主SOD和CAT的活性变化

发布时间：2018-09-19 06:58

【摘要】：目的本研究以长角血蜱和汉赛巴尔通体为对象,对长角血蜱感染汉赛巴尔通体后的生化反应和生存状态进行较为系统的研究。从生物化学水平分析蜱媒对该病原体入侵的防御反应,并分析蜱媒与病原体间的相互适应结果。通过上述研究,试图为长角血蜱与汉赛巴尔通体的媒介关系提供实验室证据,同时也为探讨蜱媒对相关病原体的免疫应答机制提供线索。方法实验采用人工注射的方式将汉赛巴尔通体菌液悬浮液注射入长角血蜱成蜱血腔内,直接观察蜱感染汉赛巴尔通体后的存活情况；在染毒后不同时间内,测定血淋巴中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性及SOD的同工酶变化；对处理后不同时间蜱的血淋巴进行革兰染色以及其与蜱残体的分离培养,观察汉赛巴尔通体在蜱体内的存活情况,通过解剖分离感染不同时间的长角血蜱内脏,观察汉赛巴尔通体在蜱体内的动态变化。另外通过研究自然状态下几种蜱感染汉赛巴尔通体的流行情况了解该种菌的分布情况。结果 1、在对长角血蜱处理后的不同时间内,注射汉赛巴尔通体组与注射生理盐水组蜱的死亡率没有统计学差异。 2、长角血蜱在接种汉赛巴尔通体后,SOD和CAT活性相对于生理盐水均在一定时间内受到抑制,虽然两种酶的表现并不一致,但在48h时均与生理盐水没有差异,这说明在48h时汉赛巴尔通体对蜱的胁迫作用已与生理盐水无异,汉赛巴尔通体对蜱可能不会产生很大的伤害。SOD同工酶谱显示在不同的时段和组织内可能有不同的同工酶参与防御反应。 3、在处理后96h内,对长角血蜱的血淋巴革兰染色,均为阳性,而血淋巴分离培养则未分离到巴尔通体。对处理后1h及24h蜱的匀浆接种均分离到汉赛巴尔通体,其他时间段未分离到。对感染后96h内蜱残体的PCR检测也全部显示阳性结果。解剖分离蜱唾液腺、中肠、卵巢及马氏管在9天内除个别外均有阳性条带。31天时汉赛巴尔通体在不同个体均有不同组织的阳性结果。 4、对蜱自然种群中汉赛巴尔通体的PCR检测,汉赛巴尔通体在海南采集的微小牛蜱中带菌率较高,其中石家庄的长角血蜱中分离培养呈阳性。结论 1、长角血蜱在受到汉赛巴尔通体的攻击之后,抗氧化酶的活性在一定时间内受到了抑制,在不同的组织中有不同的SOD同工酶参与反应。 2、汉赛巴尔通体在进入长角血蜱的血腔中之后,可以存活一段时间,并且可以通过血腔进入蜱的唾液腺,中肠、马氏管等组织内。通过上述实验室和现场调查结果,提示长角血蜱有可能成为汉赛巴尔通体的潜在传播媒介。
[Abstract]:Objective to study the biochemical reaction and survival status of Haemaphysalis longicornis and Haemaphysalis Hansebartoni after infection with Haemaphysalis longicornis. The defense response of ticks to the invasion of the pathogen was analyzed from the biochemical level, and the results of mutual adaptation between ticks and pathogens were analyzed. Through the above research, we try to provide laboratory evidence for the vector relationship between Haemaphysalis longicornis and Hansebaltonella, and also provide clues for exploring the immune response mechanism of ticks to related pathogens. Methods the suspension of Haemaphysalis longicornis was injected into the blood cavity of Haemaphysalis longicornis by artificial injection, and the survival of tick infected with Hemibaltonella longicornis was observed directly, and the survival rate was observed at different time after exposure to Haemaphysalis longicornis. The activity of superoxide dismutase (SOD) and catalase (CAT) (CAT) and the changes of SOD isozyme in hemolymph were determined, the hemolymph of ticks were stained by Gram staining at different time after treatment, and the activity of superoxide dismutase (SOD) and catalase (CAT) (CAT) were separated from ticks. To observe the survival of Hansebaltonia in ticks and to observe the dynamic changes of Hansebaltonia in ticks by dissecting and isolating the viscera of Haemaphysalis longicornis infected with different time. In addition, the distribution of several ticks infected with Hansebaltonella was studied. Results 1. After treatment with Haemaphysalis longicornis, There was no significant difference in the mortality of ticks between the two groups. 2. The activities of sod and CAT of Haemaphysalis longicornis were inhibited in a certain period of time compared with those of normal saline after inoculation of Haemaphysalis longhornii. Although the expression of the two enzymes was not consistent, there was no difference between the two enzymes at 48 h, which indicated that the stress effect of Hansebalton on ticks at 48 h was no different from that on normal saline. The isozyme spectrum of SOD showed that there might be different isozymes involved in defense response at different time periods and tissues. Hemolymph Gram staining of Haemaphysalis longicornis was positive, but Baltonella was not isolated in hemolymph isolation culture. The homogenate inoculation of ticks at 1 h and 24 h after treatment were all isolated from Hansebalton, but not from other time periods. PCR detection of tick residues within 96 hours after infection also showed positive results. Dissecting the salivary glands and midgut of ticks, Positive bands were found in ovaries and Markov tubes within 9 days except for a few. The positive results of Hensaibaltonia in different individuals were found in different individuals at day 31. 4. PCR detection of Hensaibaltonia in natural ticks population. Hensaibaltonia was found to have a high rate of carrying bacteria in the samples of Bovine ticks collected in Hainan, and the isolated culture of Haemaphysalis longicornis in Shijiazhuang was positive. Conclusion 1 the antioxidant enzyme activity of Haemaphysalis longicornis was inhibited in a certain time after being attacked by Hansebaltonella. There are different SOD isozymes involved in the reaction in different tissues. 2. After entering the blood cavity of Haemaphysalis longicornis, the Hansebartonella can survive for a period of time, and can enter the salivary gland, midgut of tick through the blood cavity. In the tissues such as the Markovian tube. The results of laboratory and field investigation indicate that Haemaphysalis longicornis may be a potential transmission medium of Hansebaltonella.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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