鸡杆菌检测方法的建立、应用及鸡输卵管囊肿的病理学研究
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摘要
鸭源鸡杆菌(Gallibacterium anatis)是感染产蛋鸡群的一种潜在病原菌,与鸡输卵管囊肿有一定的关系。本研究于2007～2010年,在对河南、山东、山西、安徽等地鸡杆菌流行病学调查的基础上,从自然病例和实验病例两方面,通过PCR方法、荧光原位杂交方法和病理组织学观察等技术手段,建立快速、特异的检测方法并观察自然感染病例、鸡杆菌单因子感染SPF蛋鸡及大肠杆菌和鸡杆菌混合感染SPF蛋鸡的病理学变化,建立病理学诊断方法,为鸡杆菌的鉴别诊断和发病机理提供一定的理论依据。研究结果表明:
1.本研究建立的鸡杆菌属特异性PCR方法和荧光原位杂交法(FISH),可对临床病料组织、分泌液及血液中的鸡杆菌进行快速、特异地检测,其特异性、敏感性均高于细菌分离法。可用于对鸡杆菌早期检测和预防。
2.鸡输卵管囊肿病例的病变主要表现在鸡的呼吸系统和生殖系统,严重影响蛋鸡的产蛋量及产蛋质量;主要剖检病变为腹膜炎和输卵管不同程度的囊肿;主要组织学变化为呼吸道黏膜上皮脱落,毛细血管充血,炎性细胞浸润,输卵管病变在膨大部和子宫的黏膜层充血、炎性细胞浸润及腺体细胞变性。
3.鸡杆菌单因子人工感染开产前后SPF鸡,引起开产前鸡群的开产日龄平均推迟7天,产蛋量下降69.7 %,劣质蛋增加70 %;开产后鸡群产蛋量下降33 %,劣质蛋增加23 %。未出现典型的剖检病变;组织学病变主要为输卵管膨大部和子宫黏膜上皮细胞增生,纤毛脱落及炎性渗出,腺体上皮细胞变性、胞核呈多形态等。
4.大肠杆菌感染开产前SPF蛋鸡后产蛋量无变化,无特征性病理组织学变化;大肠杆菌和鸡杆菌混合感染开产前SPF鸡,引起鸡群产蛋量下降87.8 %,劣质蛋增加75 %,组织学观察主要为呼吸系统和生殖系统出血、淤血,大量嗜异性白细胞浸润等。开产后SPF鸡感染大肠杆菌和/或鸡杆菌后均出现急性死亡,剖检可见心包炎、肝周炎及腹膜炎症状;呼吸系统出现严重的组织学病变。
5.混合感染对鸡群引起的病理损伤比单一感染大肠杆菌或鸡杆菌严重,鸡杆菌单因子仅影响鸡群产蛋量和产蛋质量,不引起鸡群的死亡;证实了鸡杆菌与大肠杆菌有一定的协同作用。
本项研究在国内外首次利用鸡杆菌人工感染SPF蛋鸡,并与自然感染病例进行了比较病理学研究。对鸡杆菌病的流行病学调查和鉴别诊断有重要的参考价值,并具有重要的学术意义和实用价值。
Gallibacterium anatis, which mainly infectes layers, is a latent pathogenic bacteria and has a certain relationship with oviduct cysts of chickens. From 2007 to 2010, based on the investigation of epidemiology in Henan, Anhui, Shanxi and Shandong provinces, PCR, fluorescence in situ hybridization (FISH) and histopathology were carried out to detect G.anatis from naturally and experimentally infected chickens. The detection methods developed above were rapid and specific and the pathology of chickens naturally and experimentally infected with G.anatis were observed in order to develop a pathology diagnostic method. This study provided a certain theory foundation for the differential diagnosis and the pathogensis of the disease of G. anatis. The results demonstrated that:
1. The methods of PCR and FISH developed were used to detect G. anatis in blood, excreti -on and tissues of diseased chickens. The specificity and sensitivity of the methods were higher than bacteria isolation, and could be used to early detection and epidemiological investigation.
2. The main necropsy findings were seen in the respiratory tract and genital tract of oviduct cysts, which affected the egg production and egg quality of layers; The pathological changes were peritonitis and different degree of oviduct cysts and the main histopathology were exfoliation of the mucosa epithelium cells, hyperaemia of cortex capillary and infiliatration of inflammation cells in up-respiratory tract; inflammation cells infiltrated in mucosa of magnum and uterus in oviduct, and nucleus of secretory cells broke into small pieces. The pathological changes of oviduct could be one of the important diagnosis points of the disease.
3. SPF layers artifically infected with G.anatis showed the reproductive disorders, deforme -d eggs and lowered productivity. The dominant gross pathological lesions were seen in respiratory and genital tract, and pathological changes exhibited as hyperemia, oedema, serosity and inflammation exudation. The specific histopathology in magnum and uterine part of oviduct could be one of the important diagnostic character ofthe disease. It can be affirmed, from the above results, that the G. anatis, isolated from naturally occurring cases, had a certain pathogenicity, and this study provided some differential diagnosis for complicate co-infection in clinic.
4. There was no change in egg production and quality, and no significant pathological chan -ges of 112-day layers infected with only E.coli. The combined infection with E.coli and G.anatis induced egg production dropped by 87.8%,poor quality of eggs increased by 75%; and the main histopathology changes were hemorrhage, congestion, lymphocyte cells and leukocytic infiltrate. E.coli and/or G.anatis infection caused an acute death of 164-day SPF layers; the symptoms of pericarditis, perihepatitis and peritonitis were recorded in the autopsy changes; and there was no significant histopathology expect trachea and lung.
5. The pathological damage of SPF layers infected with both E.coli and G.anatis was more severe than only G.anatis infection. G.anatis mainly affected the egg production and quality of layers, and did not cause the death of chickens. It was concluded that E.coli and G.anatis had a certain synergism.
This was the first study to induce the experimental cases infected with G. anatis and to research the pathological changes of naturally and experimentally cases. Furthermore, the results provided reference for the epidemiological investigation and differential diagnosis of G..anatis, and had very important academic and practical significance.
1.本研究建立的鸡杆菌属特异性PCR方法和荧光原位杂交法(FISH),可对临床病料组织、分泌液及血液中的鸡杆菌进行快速、特异地检测,其特异性、敏感性均高于细菌分离法。可用于对鸡杆菌早期检测和预防。
2.鸡输卵管囊肿病例的病变主要表现在鸡的呼吸系统和生殖系统,严重影响蛋鸡的产蛋量及产蛋质量;主要剖检病变为腹膜炎和输卵管不同程度的囊肿;主要组织学变化为呼吸道黏膜上皮脱落,毛细血管充血,炎性细胞浸润,输卵管病变在膨大部和子宫的黏膜层充血、炎性细胞浸润及腺体细胞变性。
3.鸡杆菌单因子人工感染开产前后SPF鸡,引起开产前鸡群的开产日龄平均推迟7天,产蛋量下降69.7 %,劣质蛋增加70 %;开产后鸡群产蛋量下降33 %,劣质蛋增加23 %。未出现典型的剖检病变;组织学病变主要为输卵管膨大部和子宫黏膜上皮细胞增生,纤毛脱落及炎性渗出,腺体上皮细胞变性、胞核呈多形态等。
4.大肠杆菌感染开产前SPF蛋鸡后产蛋量无变化,无特征性病理组织学变化;大肠杆菌和鸡杆菌混合感染开产前SPF鸡,引起鸡群产蛋量下降87.8 %,劣质蛋增加75 %,组织学观察主要为呼吸系统和生殖系统出血、淤血,大量嗜异性白细胞浸润等。开产后SPF鸡感染大肠杆菌和/或鸡杆菌后均出现急性死亡,剖检可见心包炎、肝周炎及腹膜炎症状;呼吸系统出现严重的组织学病变。
5.混合感染对鸡群引起的病理损伤比单一感染大肠杆菌或鸡杆菌严重,鸡杆菌单因子仅影响鸡群产蛋量和产蛋质量,不引起鸡群的死亡;证实了鸡杆菌与大肠杆菌有一定的协同作用。
本项研究在国内外首次利用鸡杆菌人工感染SPF蛋鸡,并与自然感染病例进行了比较病理学研究。对鸡杆菌病的流行病学调查和鉴别诊断有重要的参考价值,并具有重要的学术意义和实用价值。
Gallibacterium anatis, which mainly infectes layers, is a latent pathogenic bacteria and has a certain relationship with oviduct cysts of chickens. From 2007 to 2010, based on the investigation of epidemiology in Henan, Anhui, Shanxi and Shandong provinces, PCR, fluorescence in situ hybridization (FISH) and histopathology were carried out to detect G.anatis from naturally and experimentally infected chickens. The detection methods developed above were rapid and specific and the pathology of chickens naturally and experimentally infected with G.anatis were observed in order to develop a pathology diagnostic method. This study provided a certain theory foundation for the differential diagnosis and the pathogensis of the disease of G. anatis. The results demonstrated that:
1. The methods of PCR and FISH developed were used to detect G. anatis in blood, excreti -on and tissues of diseased chickens. The specificity and sensitivity of the methods were higher than bacteria isolation, and could be used to early detection and epidemiological investigation.
2. The main necropsy findings were seen in the respiratory tract and genital tract of oviduct cysts, which affected the egg production and egg quality of layers; The pathological changes were peritonitis and different degree of oviduct cysts and the main histopathology were exfoliation of the mucosa epithelium cells, hyperaemia of cortex capillary and infiliatration of inflammation cells in up-respiratory tract; inflammation cells infiltrated in mucosa of magnum and uterus in oviduct, and nucleus of secretory cells broke into small pieces. The pathological changes of oviduct could be one of the important diagnosis points of the disease.
3. SPF layers artifically infected with G.anatis showed the reproductive disorders, deforme -d eggs and lowered productivity. The dominant gross pathological lesions were seen in respiratory and genital tract, and pathological changes exhibited as hyperemia, oedema, serosity and inflammation exudation. The specific histopathology in magnum and uterine part of oviduct could be one of the important diagnostic character ofthe disease. It can be affirmed, from the above results, that the G. anatis, isolated from naturally occurring cases, had a certain pathogenicity, and this study provided some differential diagnosis for complicate co-infection in clinic.
4. There was no change in egg production and quality, and no significant pathological chan -ges of 112-day layers infected with only E.coli. The combined infection with E.coli and G.anatis induced egg production dropped by 87.8%,poor quality of eggs increased by 75%; and the main histopathology changes were hemorrhage, congestion, lymphocyte cells and leukocytic infiltrate. E.coli and/or G.anatis infection caused an acute death of 164-day SPF layers; the symptoms of pericarditis, perihepatitis and peritonitis were recorded in the autopsy changes; and there was no significant histopathology expect trachea and lung.
5. The pathological damage of SPF layers infected with both E.coli and G.anatis was more severe than only G.anatis infection. G.anatis mainly affected the egg production and quality of layers, and did not cause the death of chickens. It was concluded that E.coli and G.anatis had a certain synergism.
This was the first study to induce the experimental cases infected with G. anatis and to research the pathological changes of naturally and experimentally cases. Furthermore, the results provided reference for the epidemiological investigation and differential diagnosis of G..anatis, and had very important academic and practical significance.
引文
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