坦布苏病毒对雏鹅的致病性研究
发布时间：2019-05-17 12:49
【摘要】：自2010年4月起,我国多地蛋鸭、种鹅养殖场相继爆发了一种以产蛋量严重下降为主要临床症状的新型传染病,经分离鉴定确定病原为坦布苏病毒(Tembusu virus,TMUV)。坦布苏病毒主要感染鸭,但也能感染鹅,造成鹅的发病和死亡。种鹅感染TMUV后严重影响种鹅产蛋量,雏鹅感染坦布苏病毒发病严重,生长迟缓,最终可导致部分发病严重雏鹅死亡,已经给我国养鹅业带来巨大的经济损失。本研究用实验室分离到的鸭源坦布苏病毒分别对5日龄和20日龄的雏鹅进行人工感染试验,研究该病毒对雏鹅的致病性,旨在为今后鹅坦布苏病毒感染的防控提供理论依据。100只1日龄健康雏鹅平均分为5组,每组20只。1-4组为实验组,5组为对照组。1-2组于5日龄、3-4组于20日龄分别经静脉和点眼滴鼻人工接种TMUV SDSG株(ELD50为10-2.17/0.2mL),5组接种等剂量的生理盐水。观察鹅的发病情况和临床症状,在攻毒后第3d、6d、9d、12d于各组随机选取3只雏鹅,采血并剖杀,观察剖检变化,采集脑、肝脏、肺脏、胰脏、脾脏等,进行病理组织学观察、载毒量分析和血液细胞因子测定。结果显示:发病鹅临床症状主要表现为,5日龄静脉注射攻毒组自攻毒后第4d开始发病,点眼滴鼻组自攻毒后第5d开始发病,发病鹅出现精神沉郁,厌食,排白绿色、棕色稀便,两腿无力,站立不稳,颤抖,生长迟缓,伴有腹部朝上两腿挣扎呈游泳状,角弓反张直至死亡等症状,偶见静脉注射组发病鹅出现头颈震颤神经症状。自攻毒后第5d静脉注射组有病鹅出现死亡,攻毒后第6d点眼滴鼻组开始出现死亡,发病时间集中在攻毒后4-8d,死亡时间集中在攻毒后的5-8d,从第9d开始临床症状逐渐减轻。20日龄攻毒组自攻毒5d后开始出现食欲下降,精神不振,排黄绿色稀便,病程维持3d左右,攻毒后8-9d临床症状消失,采食量回升。分别在攻毒后第3d、6d、9d、12d测定攻毒鹅和对照鹅的体重,5日龄攻毒组较20日龄攻毒组发病鹅体重下降更为明显。病死鹅剖检病变为多个组织器官的出血和坏死,包括脑膜充血、出血、脑水肿;心冠脂肪出血、心内膜出血、心肌坏死;肺水肿、出血、淤血;腺胃乳头轻度出血;肝脏出血、坏死;胰腺水肿、坏死;脾脏肿大、出血;肾脏出血、水肿等病变。病理组织学变化主要包括,脑血管周围间隙变大水肿、脑膜有大量炎性细胞浸润、充血、脑间质散在小胶质细胞增生;肝、胰腺、肾脏的腺管上皮细胞凋亡、坏死,腺管间隙扩张淤血;脾脏大量淋巴细胞凋亡空泡化;心肌断裂、变性、坏死、心肌纤维间距变大并伴有炎性细胞浸润;肺出血、淤血等病变明显。实时荧光定量PCR检测结果显示,胰腺和脑是病毒载量最高且持续时间最长的器官,心、脾、肾、肠、胸腺、法氏囊、腺胃含量则次之,相对表达量较低的器官主要是肝和肺。攻毒3d后各组织均可检测到病毒,攻毒6 d后病毒载量达到高峰,随后病毒含量逐渐降低,攻毒后第12d肝、肺、胸腺和腺胃检测不到病毒。比较5日龄攻毒组和20日龄攻毒组各组织器官的病毒载量,5日龄攻毒组是20日龄攻毒组的10倍以上。细胞因子测定结果显示:测定IL-4,IFN-γ结果显示,细胞因子水平呈现先下降后上升的总趋势。人工感染雏鹅第3d,IL-4、IFN-γ的含量与对照组相比无明显差异性;第6-9d,IL-4、IFN-γ含量低于对照组,自第9d开始IL-4、IFN-γ的含量开始逐渐上升,第12d可见IL-4、IFN-γ的含量高于对照组。抗体水平测定结果显示:在攻毒后第3d未检测到坦布苏病毒抗体,第6d可检测到抗体,随后抗体水平呈上升趋势直至第12d依然上升。综上所述,坦布苏病毒对雏鹅致病性的显著,5日龄试验组临床症状、剖检病变和病理组织学变化明显显著于20日龄试验组。因此,在生产中,应做好鹅坦布苏病毒感染的防控。
[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.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.33
本文编号：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.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.33
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相关期刊论文 前4条
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