【摘要】：目的 本研究以長角血蜱和漢賽巴爾通體為對象,對長角血蜱感染漢賽巴爾通體后的生化反應和生存狀態(tài)進行較為系統(tǒng)的研究。從生物化學水平分析蜱媒對該病原體入侵的防御反應,并分析蜱媒與病原體間的相互適應結果。通過上述研究,試圖為長角血蜱與漢賽巴爾通體的媒介關系提供實驗室證據,同時也為探討蜱媒對相關病原體的免疫應答機制提供線索。 方法 實驗采用人工注射的方式將漢賽巴爾通體菌液懸浮液注射入長角血蜱成蜱血腔內,直接觀察蜱感染漢賽巴爾通體后的存活情況；在染毒后不同時間內,測定血淋巴中超氧化物歧化酶(SOD)和過氧化氫酶(CAT)的活性及SOD的同工酶變化；對處理后不同時間蜱的血淋巴進行革蘭染色以及其與蜱殘體的分離培養(yǎng),觀察漢賽巴爾通體在蜱體內的存活情況,通過解剖分離感染不同時間的長角血蜱內臟,觀察漢賽巴爾通體在蜱體內的動態(tài)變化。另外通過研究自然狀態(tài)下幾種蜱感染漢賽巴爾通體的流行情況了解該種菌的分布情況。 結果 1、在對長角血蜱處理后的不同時間內,注射漢賽巴爾通體組與注射生理鹽水組蜱的死亡率沒有統(tǒng)計學差異。 2、長角血蜱在接種漢賽巴爾通體后,SOD和CAT活性相對于生理鹽水均在一定時間內受到抑制,雖然兩種酶的表現并不一致,但在48h時均與生理鹽水沒有差異,這說明在48h時漢賽巴爾通體對蜱的脅迫作用已與生理鹽水無異,漢賽巴爾通體對蜱可能不會產生很大的傷害。SOD同工酶譜顯示在不同的時段和組織內可能有不同的同工酶參與防御反應。 3、在處理后96h內,對長角血蜱的血淋巴革蘭染色,均為陽性,而血淋巴分離培養(yǎng)則未分離到巴爾通體。對處理后1h及24h蜱的勻漿接種均分離到漢賽巴爾通體,其他時間段未分離到。對感染后96h內蜱殘體的PCR檢測也全部顯示陽性結果。解剖分離蜱唾液腺、中腸、卵巢及馬氏管在9天內除個別外均有陽性條帶。31天時漢賽巴爾通體在不同個體均有不同組織的陽性結果。 4、對蜱自然種群中漢賽巴爾通體的PCR檢測,漢賽巴爾通體在海南采集的微小牛蜱中帶菌率較高,其中石家莊的長角血蜱中分離培養(yǎng)呈陽性。 結論 1、長角血蜱在受到漢賽巴爾通體的攻擊之后,抗氧化酶的活性在一定時間內受到了抑制,在不同的組織中有不同的SOD同工酶參與反應。 2、漢賽巴爾通體在進入長角血蜱的血腔中之后,可以存活一段時間,并且可以通過血腔進入蜱的唾液腺,中腸、馬氏管等組織內。 通過上述實驗室和現場調查結果,提示長角血蜱有可能成為漢賽巴爾通體的潛在傳播媒介。
[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.
【學位授予單位】：鄭州大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R363

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