坦布蘇病毒對雛鵝的致病性研究
發(fā)布時間：2019-05-17 12:49
【摘要】：自2010年4月起,我國多地蛋鴨、種鵝養(yǎng)殖場相繼爆發(fā)了一種以產(chǎn)蛋量嚴重下降為主要臨床癥狀的新型傳染病,經(jīng)分離鑒定確定病原為坦布蘇病毒(Tembusu virus,TMUV)。坦布蘇病毒主要感染鴨,但也能感染鵝,造成鵝的發(fā)病和死亡。種鵝感染TMUV后嚴重影響種鵝產(chǎn)蛋量,雛鵝感染坦布蘇病毒發(fā)病嚴重,生長遲緩,最終可導致部分發(fā)病嚴重雛鵝死亡,已經(jīng)給我國養(yǎng)鵝業(yè)帶來巨大的經(jīng)濟損失。本研究用實驗室分離到的鴨源坦布蘇病毒分別對5日齡和20日齡的雛鵝進行人工感染試驗,研究該病毒對雛鵝的致病性,旨在為今后鵝坦布蘇病毒感染的防控提供理論依據(jù)。100只1日齡健康雛鵝平均分為5組,每組20只。1-4組為實驗組,5組為對照組。1-2組于5日齡、3-4組于20日齡分別經(jīng)靜脈和點眼滴鼻人工接種TMUV SDSG株(ELD50為10-2.17/0.2mL),5組接種等劑量的生理鹽水。觀察鵝的發(fā)病情況和臨床癥狀,在攻毒后第3d、6d、9d、12d于各組隨機選取3只雛鵝,采血并剖殺,觀察剖檢變化,采集腦、肝臟、肺臟、胰臟、脾臟等,進行病理組織學觀察、載毒量分析和血液細胞因子測定。結果顯示:發(fā)病鵝臨床癥狀主要表現(xiàn)為,5日齡靜脈注射攻毒組自攻毒后第4d開始發(fā)病,點眼滴鼻組自攻毒后第5d開始發(fā)病,發(fā)病鵝出現(xiàn)精神沉郁,厭食,排白綠色、棕色稀便,兩腿無力,站立不穩(wěn),顫抖,生長遲緩,伴有腹部朝上兩腿掙扎呈游泳狀,角弓反張直至死亡等癥狀,偶見靜脈注射組發(fā)病鵝出現(xiàn)頭頸震顫神經(jīng)癥狀。自攻毒后第5d靜脈注射組有病鵝出現(xiàn)死亡,攻毒后第6d點眼滴鼻組開始出現(xiàn)死亡,發(fā)病時間集中在攻毒后4-8d,死亡時間集中在攻毒后的5-8d,從第9d開始臨床癥狀逐漸減輕。20日齡攻毒組自攻毒5d后開始出現(xiàn)食欲下降,精神不振,排黃綠色稀便,病程維持3d左右,攻毒后8-9d臨床癥狀消失,采食量回升。分別在攻毒后第3d、6d、9d、12d測定攻毒鵝和對照鵝的體重,5日齡攻毒組較20日齡攻毒組發(fā)病鵝體重下降更為明顯。病死鵝剖檢病變?yōu)槎鄠�組織器官的出血和壞死,包括腦膜充血、出血、腦水腫;心冠脂肪出血、心內膜出血、心肌壞死;肺水腫、出血、淤血;腺胃乳頭輕度出血;肝臟出血、壞死;胰腺水腫、壞死;脾臟腫大、出血;腎臟出血、水腫等病變。病理組織學變化主要包括,腦血管周圍間隙變大水腫、腦膜有大量炎性細胞浸潤、充血、腦間質散在小膠質細胞增生;肝、胰腺、腎臟的腺管上皮細胞凋亡、壞死,腺管間隙擴張淤血;脾臟大量淋巴細胞凋亡空泡化;心肌斷裂、變性、壞死、心肌纖維間距變大并伴有炎性細胞浸潤;肺出血、淤血等病變明顯。實時熒光定量PCR檢測結果顯示,胰腺和腦是病毒載量最高且持續(xù)時間最長的器官,心、脾、腎、腸、胸腺、法氏囊、腺胃含量則次之,相對表達量較低的器官主要是肝和肺。攻毒3d后各組織均可檢測到病毒,攻毒6 d后病毒載量達到高峰,隨后病毒含量逐漸降低,攻毒后第12d肝、肺、胸腺和腺胃檢測不到病毒。比較5日齡攻毒組和20日齡攻毒組各組織器官的病毒載量,5日齡攻毒組是20日齡攻毒組的10倍以上。細胞因子測定結果顯示:測定IL-4,IFN-γ結果顯示,細胞因子水平呈現(xiàn)先下降后上升的總趨勢。人工感染雛鵝第3d,IL-4、IFN-γ的含量與對照組相比無明顯差異性;第6-9d,IL-4、IFN-γ含量低于對照組,自第9d開始IL-4、IFN-γ的含量開始逐漸上升,第12d可見IL-4、IFN-γ的含量高于對照組。抗體水平測定結果顯示:在攻毒后第3d未檢測到坦布蘇病毒抗體,第6d可檢測到抗體,隨后抗體水平呈上升趨勢直至第12d依然上升。綜上所述,坦布蘇病毒對雛鵝致病性的顯著,5日齡試驗組臨床癥狀、剖檢病變和病理組織學變化明顯顯著于20日齡試驗組。因此,在生產(chǎn)中,應做好鵝坦布蘇病毒感染的防控。
[Abstract]:Since April,2010, a new type of infectious disease, which is a major clinical symptom, has been developed in China's laying ducks and breeding farms, and it is determined that the pathogen is Tembusu virus (TMUV). The Tambsu virus is mainly infected with duck, but can also be infected with the goose, causing the morbidity and death of the goose. The infection of the goose with TMV seriously affects the egg production of the goose, the infection of the gosling is serious and the growth is slow, which can lead to the death of some serious goslings, which has brought great economic losses to the goose-raising industry in China. In this study, the experiment of artificial infection of the 5-day-old and 20-day-old goslings was carried out by using the duck-source, which was separated from the laboratory, and the pathogenicity of the virus to the goose was studied. The aim of this study was to provide a theoretical basis for the prevention and control of the infection of the goose in the future. 3-4 groups were inoculated with TMUV SDSG (10-2.17/ 0.2 mL) and 5 groups of normal saline at 20-day-old. The morbidity and clinical symptoms of the goose were observed. Three young geese were randomly selected at the 3rd day, the 6th day, the 9th day and the 12th day after the challenge. The changes of the cross-section were observed, the brain, the liver, the lung, the pancreas, the spleen and the like were collected. Analysis of drug-carrying amount and determination of blood cytokine. The results showed that the clinical symptoms of the goose were mainly characterized by the onset of the fourth day after the self-tapping of the 5-day-old group of the challenge group, and the onset of the fifth day after the self-tapping of the group of the eye drops. The disease of the goose was depressed, the anorexia, the white-green, the brown and the weak, the legs were weak, the standing was unstable, and the shaking, The growth retardation, with the abdomen facing upward, the legs struggle to swim, the angle bow to the back to death, and so on, occasionally see the onset of the IV group, the disease of the head and neck tremors. In the fifth day after self-tapping, there was a dead goose in the IV group. At the 6th day after the challenge, the group started to die, and the time of the attack was 4-8 days after challenge, and the time of death was concentrated on 5-8 days after challenge. The clinical symptoms were gradually reduced from the 9th day. The 20-day-old challenge group began to show a decrease in appetite after 5 days of challenge, and the symptoms disappeared after the challenge. The clinical symptoms of 8-9 days after challenge disappeared and the feed intake returned. The weight of the challenge goose and the control goose was determined on the 3rd day, the 6th day, the 9th day and the 12th day after the challenge, and the weight of the goose was more obvious in the 5-day-old challenge group than that of the 20-day-old attack group. The disease is the bleeding and necrosis of a plurality of tissue organs, including meninges congestion, hemorrhage, cerebral edema, coronary heart crown fat hemorrhage, endocardium hemorrhage, myocardial necrosis, pulmonary edema, hemorrhage, blood stasis, slight bleeding of the gland and stomach papilla, liver hemorrhage, necrosis, pancreatic edema, necrosis, and the like. Splenomegaly, hemorrhage; renal hemorrhage, edema, etc. The pathological changes mainly include, the peripheral clearance of the cerebral vessel is enlarged, the meninges have a large number of inflammatory cell infiltration, hyperemia, and the interstitial powder of the brain is in the proliferation of the microglia; the cells of the glandular tube of the liver, the pancreas and the kidney are apoptosis, necrosis, and the gap of the glandular tube is expanded and blood stasis; A large number of lymphocytes in the spleen were vacuolated; the myocardial rupture, degeneration, necrosis, the distance between the cardiac muscle fibers and the infiltration of inflammatory cells, the pulmonary hemorrhage, the blood stasis, and the like were obvious. The results of real-time fluorescence quantitative PCR showed that the pancreas and brain were the most viral load and the longest duration of organ, heart, spleen, kidney, intestine, thymus, bursa of Fabricius, and glandular stomach, and the organ with lower relative expression was mainly liver and lung. After 3 days of challenge, the virus was detected and the viral load reached the peak after 6 days of challenge, and the virus content was gradually reduced, and the virus was not detected in the liver, lung, thymus and glandular stomach at the 12th day after challenge. The 5-day-old challenge group was more than 10-fold higher than that of the 20-day-old attack group, compared with the viral load of each tissue organ in the 5-day-old attack group and the 20-day-old attack group. The results of the measurement of cytokines showed that the results of IL-4 and IFN-PCR showed that the level of cytokines decreased first and then increased. The content of IL-4 and IFN-1 was lower than that in the control group. The content of IL-4 and IFN-1 in the first day of day 9 was higher than that of the control group, and the content of IFN-1 was higher than that of the control group. The results of the antibody level determination showed that the antibody was not detected on the 3rd day after challenge, and the antibody was detected on the 6th day, and then the antibody level was on the rise until the 12th day. In conclusion, the clinical symptoms, cross-section and pathological changes of the test group in the 5-day-old group were significantly higher than that of the 20-day-old test group. Therefore, in the production, it is necessary to do the prevention and control of the infection of the goose.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.33
本文編號：2479103
[Abstract]:Since April,2010, a new type of infectious disease, which is a major clinical symptom, has been developed in China's laying ducks and breeding farms, and it is determined that the pathogen is Tembusu virus (TMUV). The Tambsu virus is mainly infected with duck, but can also be infected with the goose, causing the morbidity and death of the goose. The infection of the goose with TMV seriously affects the egg production of the goose, the infection of the gosling is serious and the growth is slow, which can lead to the death of some serious goslings, which has brought great economic losses to the goose-raising industry in China. In this study, the experiment of artificial infection of the 5-day-old and 20-day-old goslings was carried out by using the duck-source, which was separated from the laboratory, and the pathogenicity of the virus to the goose was studied. The aim of this study was to provide a theoretical basis for the prevention and control of the infection of the goose in the future. 3-4 groups were inoculated with TMUV SDSG (10-2.17/ 0.2 mL) and 5 groups of normal saline at 20-day-old. The morbidity and clinical symptoms of the goose were observed. Three young geese were randomly selected at the 3rd day, the 6th day, the 9th day and the 12th day after the challenge. The changes of the cross-section were observed, the brain, the liver, the lung, the pancreas, the spleen and the like were collected. Analysis of drug-carrying amount and determination of blood cytokine. The results showed that the clinical symptoms of the goose were mainly characterized by the onset of the fourth day after the self-tapping of the 5-day-old group of the challenge group, and the onset of the fifth day after the self-tapping of the group of the eye drops. The disease of the goose was depressed, the anorexia, the white-green, the brown and the weak, the legs were weak, the standing was unstable, and the shaking, The growth retardation, with the abdomen facing upward, the legs struggle to swim, the angle bow to the back to death, and so on, occasionally see the onset of the IV group, the disease of the head and neck tremors. In the fifth day after self-tapping, there was a dead goose in the IV group. At the 6th day after the challenge, the group started to die, and the time of the attack was 4-8 days after challenge, and the time of death was concentrated on 5-8 days after challenge. The clinical symptoms were gradually reduced from the 9th day. The 20-day-old challenge group began to show a decrease in appetite after 5 days of challenge, and the symptoms disappeared after the challenge. The clinical symptoms of 8-9 days after challenge disappeared and the feed intake returned. The weight of the challenge goose and the control goose was determined on the 3rd day, the 6th day, the 9th day and the 12th day after the challenge, and the weight of the goose was more obvious in the 5-day-old challenge group than that of the 20-day-old attack group. The disease is the bleeding and necrosis of a plurality of tissue organs, including meninges congestion, hemorrhage, cerebral edema, coronary heart crown fat hemorrhage, endocardium hemorrhage, myocardial necrosis, pulmonary edema, hemorrhage, blood stasis, slight bleeding of the gland and stomach papilla, liver hemorrhage, necrosis, pancreatic edema, necrosis, and the like. Splenomegaly, hemorrhage; renal hemorrhage, edema, etc. The pathological changes mainly include, the peripheral clearance of the cerebral vessel is enlarged, the meninges have a large number of inflammatory cell infiltration, hyperemia, and the interstitial powder of the brain is in the proliferation of the microglia; the cells of the glandular tube of the liver, the pancreas and the kidney are apoptosis, necrosis, and the gap of the glandular tube is expanded and blood stasis; A large number of lymphocytes in the spleen were vacuolated; the myocardial rupture, degeneration, necrosis, the distance between the cardiac muscle fibers and the infiltration of inflammatory cells, the pulmonary hemorrhage, the blood stasis, and the like were obvious. The results of real-time fluorescence quantitative PCR showed that the pancreas and brain were the most viral load and the longest duration of organ, heart, spleen, kidney, intestine, thymus, bursa of Fabricius, and glandular stomach, and the organ with lower relative expression was mainly liver and lung. After 3 days of challenge, the virus was detected and the viral load reached the peak after 6 days of challenge, and the virus content was gradually reduced, and the virus was not detected in the liver, lung, thymus and glandular stomach at the 12th day after challenge. The 5-day-old challenge group was more than 10-fold higher than that of the 20-day-old attack group, compared with the viral load of each tissue organ in the 5-day-old attack group and the 20-day-old attack group. The results of the measurement of cytokines showed that the results of IL-4 and IFN-PCR showed that the level of cytokines decreased first and then increased. The content of IL-4 and IFN-1 was lower than that in the control group. The content of IL-4 and IFN-1 in the first day of day 9 was higher than that of the control group, and the content of IFN-1 was higher than that of the control group. The results of the antibody level determination showed that the antibody was not detected on the 3rd day after challenge, and the antibody was detected on the 6th day, and then the antibody level was on the rise until the 12th day. In conclusion, the clinical symptoms, cross-section and pathological changes of the test group in the 5-day-old group were significantly higher than that of the 20-day-old test group. Therefore, in the production, it is necessary to do the prevention and control of the infection of the goose.
【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.33
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