奶山羊干酪性淋巴結(jié)炎病原的分離鑒定及血清學(xué)調(diào)查
發(fā)布時間：2018-09-18 19:11
【摘要】：奶山羊規(guī)�；B(yǎng)殖已成為我國奶山羊產(chǎn)業(yè)的主要發(fā)展方向,由于羊奶價格的競爭優(yōu)勢,養(yǎng)殖奶山羊的地區(qū)也越來越多,如河南、黑龍江、福建、貴州等省份。在我國奶山羊主要養(yǎng)殖省份陜西和云南,規(guī)�；B(yǎng)殖已經(jīng)被養(yǎng)殖者普遍接受。但奶山羊傳染病至今仍是嚴重制約我國奶山羊產(chǎn)業(yè)規(guī)�；】蛋l(fā)展的主要因素之一,尤其是難以治愈的奶山羊干酪性淋巴結(jié)炎(又名山羊偽結(jié)核病,Caseous lymphadenitis(CLA))�；疾∧躺窖虿粌H出現(xiàn)體重下降、生產(chǎn)性能差、繁殖功能障礙等疾病癥狀。尤為嚴重的是,泌乳羊乳汁中含有一定數(shù)量的病原菌,從而對喜食生羊乳的消費者健康造成巨大隱患。奶山羊干酪性淋巴結(jié)炎在我國各地很多養(yǎng)殖場都可看到,但更為準(zhǔn)確地流行病學(xué)調(diào)查數(shù)據(jù)仍是空白,這也是本研究要解決的主要問題之一。本研究選取陜西富、隴縣和云南瀘西的奶山羊養(yǎng)殖場,采集疑似山羊干酪性淋巴結(jié)炎病羊膿腫膿汁,采用細菌分離培養(yǎng)、顯微鏡觀察、生化試驗、協(xié)同溶血試驗、PCR、動物致病性試驗等方法對病原菌進行分離鑒定;采用間接ELISA對采集的奶山羊血清進行CLA抗體測定。研究結(jié)果如下:1.從陜西富平、隴縣和云南瀘西奶山羊羊群采集的樣品中共分離到3株菌株。分離菌株在普通瓊脂平板上生長貧瘠,形成灰白色、圓形、較干燥小菌落;麥康凱瓊脂平板上不生長。鮮血瓊脂平板上可見灰白色、不透明、同心圓狀、邊緣不齊、有狹窄β溶血環(huán)菌落,傳代后溶血現(xiàn)象消失。LB液體培養(yǎng)基中表面形成白色菌膜,管底有顆粒狀、絮狀沉淀。細菌鏡檢革蘭氏染色陽性,無莢膜,無芽孢,不運動,多呈棒狀或球桿狀。細菌生化特性與《伯杰細菌鑒定手冊》所述偽結(jié)核棒狀桿菌生化特性基本一致。2.在綿羊鮮血平板上,分離菌株能與馬紅球菌呈現(xiàn)特征性協(xié)同溶血現(xiàn)象。3.分子生物學(xué)檢測顯示3株菌株與Gen Bank中已收錄的偽結(jié)核棒狀桿菌參考株親緣關(guān)系97%以上。4.Balb/c小鼠致病性試驗和奶山羊回歸試驗證實分離菌株具有很強的致病性。5.陜西富平、隴縣和云南瀘西3個地區(qū)奶山羊干酪性淋巴結(jié)炎血清抗體陽性率為59.92%。其中,富平、隴縣和瀘西奶山羊干酪性淋巴結(jié)炎血清抗體陽性率分別為:71.50%、67.50%和25.47%;半放牧半舍飼養(yǎng)殖、集約化養(yǎng)殖和合作社養(yǎng)殖下奶山羊干酪性淋巴結(jié)炎血清抗體陽性率分別為:25.47%、67.66%和75%。6.經(jīng)兩樣本百分數(shù)檢驗發(fā)現(xiàn):成年奶山羊比其他生理階段的山羊?qū)δ躺窖蚋衫倚粤馨徒Y(jié)炎更易感;與其他地區(qū)奶山羊相比,云南瀘西奶山羊?qū)ι窖蚋衫倚粤馨徒Y(jié)炎最不易感。在不同養(yǎng)殖模式下,合作社養(yǎng)殖奶山羊?qū)ι窖蚋衫倚粤馨徒Y(jié)炎最易感。結(jié)論:1.成功分離3株山羊干酪性淋巴結(jié)炎病原菌;2.明確陜西富平、隴縣和云南瀘西奶山羊CLA現(xiàn)行流行情況。
[Abstract]:Large-scale breeding of dairy goats has become the main development direction of dairy goat industry in China. Due to the competitive advantage of goat milk prices, there are more and more areas where milk goats are raised, such as Henan, Heilongjiang, Fujian, Guizhou and other provinces. In Shaanxi and Yunnan, the main provinces of dairy goat farming in China, large-scale farming has been widely accepted by farmers. However, dairy goat infectious disease is still one of the major factors restricting the healthy development of dairy goat industry, especially caseous lymphadenitis (CLA)., which is difficult to cure. Sick dairy goats not only have weight loss, poor production performance, reproductive dysfunction and other disease symptoms. Especially, the milk of lactating sheep contains a certain number of pathogens, which causes a great danger to the health of consumers who like to eat raw goat milk. Cheese lymphadenitis of dairy goats can be seen in many farms in China, but more accurate epidemiological survey data are still blank, which is one of the main problems to be solved in this study. In this study, milk goat farms in Fu, Longxian and Luxi, Yunnan Province were selected to collect abscess from goat abscess suspected to be caseous lymphadenitis. Bacteria isolation culture, microscopic observation and biochemical test were used. The pathogenic bacteria were isolated and identified by synergistic hemolysis test and animal pathogenicity test. Indirect ELISA was used to detect the CLA antibody in the collected milk goat serum. The results are as follows: 1. Three strains were isolated from sheep samples collected from sheep in Fuping, Longxian and Luxi, Yunnan. The isolated strains grew barren on the ordinary Agar plate and formed a small colony with gray white round and dry while that of wheat Kang Kai did not grow on the Agar plate. On the blood Agar plate, there were grayish white, opaque, concentric circle, irregular edge, narrow 尾 hemolytic circle colony. After passage, the hemolysis phenomenon disappeared. The surface of the liquid culture medium of LB formed white bacterial membrane, and the bottom of the tube was granular and flocculent. Microscopical examination showed Gram-positive staining, no capsule, no spores, no movement, mostly stick or club shape. The biochemical characteristics of bacteria are basically consistent with those of Corynebacterium pseudotuberculosis described in Berger's Manual of bacterial Identification. On the sheep blood plate, the isolated strain showed characteristic synergistic hemolysis with Erythrococcus equisetii. 3. Molecular biology analysis showed that the relationship between the three strains and the reference strain of Corynebacterium pseudotuberculosis in Gen Bank was more than 97%. 4. The pathogenicity test of Balb / c mice and the regression test of dairy goat confirmed that the isolated strains had strong pathogenicity. 5. The positive rate of serum antibodies against caseous lymphadenitis of dairy goats in three areas of Fuping, Longxian and Luxi of Yunnan was 59.92. The positive rates of antibodies against caseous lymphadenitis in Fuping, Longxian and Luxi dairy goats were 67.50% and 25.47%, respectively. The positive rates of serum antibodies against caseous lymphadenitis of dairy goats in intensive farming and cooperative farming were 67.66% and 75.6%, respectively. It was found that adult dairy goats were more susceptible to cheesy lymphadenitis than those in other physiological stages, and Luxi dairy goats were the least susceptible to caseous lymphadenitis compared with other dairy goats. In different breeding modes, dairy goats were most susceptible to caseous lymphadenitis in cooperative farming. Conclusion 1. Three strains of caseous lymphadenitis were isolated successfully. To determine the current prevalence of CLA in Fuping, Longxian and Luxi dairy goats in Yunnan.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S858.27
本文編號：2248841
[Abstract]:Large-scale breeding of dairy goats has become the main development direction of dairy goat industry in China. Due to the competitive advantage of goat milk prices, there are more and more areas where milk goats are raised, such as Henan, Heilongjiang, Fujian, Guizhou and other provinces. In Shaanxi and Yunnan, the main provinces of dairy goat farming in China, large-scale farming has been widely accepted by farmers. However, dairy goat infectious disease is still one of the major factors restricting the healthy development of dairy goat industry, especially caseous lymphadenitis (CLA)., which is difficult to cure. Sick dairy goats not only have weight loss, poor production performance, reproductive dysfunction and other disease symptoms. Especially, the milk of lactating sheep contains a certain number of pathogens, which causes a great danger to the health of consumers who like to eat raw goat milk. Cheese lymphadenitis of dairy goats can be seen in many farms in China, but more accurate epidemiological survey data are still blank, which is one of the main problems to be solved in this study. In this study, milk goat farms in Fu, Longxian and Luxi, Yunnan Province were selected to collect abscess from goat abscess suspected to be caseous lymphadenitis. Bacteria isolation culture, microscopic observation and biochemical test were used. The pathogenic bacteria were isolated and identified by synergistic hemolysis test and animal pathogenicity test. Indirect ELISA was used to detect the CLA antibody in the collected milk goat serum. The results are as follows: 1. Three strains were isolated from sheep samples collected from sheep in Fuping, Longxian and Luxi, Yunnan. The isolated strains grew barren on the ordinary Agar plate and formed a small colony with gray white round and dry while that of wheat Kang Kai did not grow on the Agar plate. On the blood Agar plate, there were grayish white, opaque, concentric circle, irregular edge, narrow 尾 hemolytic circle colony. After passage, the hemolysis phenomenon disappeared. The surface of the liquid culture medium of LB formed white bacterial membrane, and the bottom of the tube was granular and flocculent. Microscopical examination showed Gram-positive staining, no capsule, no spores, no movement, mostly stick or club shape. The biochemical characteristics of bacteria are basically consistent with those of Corynebacterium pseudotuberculosis described in Berger's Manual of bacterial Identification. On the sheep blood plate, the isolated strain showed characteristic synergistic hemolysis with Erythrococcus equisetii. 3. Molecular biology analysis showed that the relationship between the three strains and the reference strain of Corynebacterium pseudotuberculosis in Gen Bank was more than 97%. 4. The pathogenicity test of Balb / c mice and the regression test of dairy goat confirmed that the isolated strains had strong pathogenicity. 5. The positive rate of serum antibodies against caseous lymphadenitis of dairy goats in three areas of Fuping, Longxian and Luxi of Yunnan was 59.92. The positive rates of antibodies against caseous lymphadenitis in Fuping, Longxian and Luxi dairy goats were 67.50% and 25.47%, respectively. The positive rates of serum antibodies against caseous lymphadenitis of dairy goats in intensive farming and cooperative farming were 67.66% and 75.6%, respectively. It was found that adult dairy goats were more susceptible to cheesy lymphadenitis than those in other physiological stages, and Luxi dairy goats were the least susceptible to caseous lymphadenitis compared with other dairy goats. In different breeding modes, dairy goats were most susceptible to caseous lymphadenitis in cooperative farming. Conclusion 1. Three strains of caseous lymphadenitis were isolated successfully. To determine the current prevalence of CLA in Fuping, Longxian and Luxi dairy goats in Yunnan.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S858.27
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