鸡球虫的分离鉴定及防治鸡球虫病中西药复方制剂的研究
|
摘要
鸡球虫病是由艾美耳属一种或多种球虫寄生于鸡体内所引起的呈世界性分布的原虫病,该病对雏鸡的危害十分严重,给养鸡业造成了巨大的经济损失。鸡球虫种类多,且临床多呈混合感染,其中致病性最强的是柔嫩艾美耳球虫和毒害艾美耳球虫。本研究从安徽部分地区鸡的粪样内分离球虫卵囊,并采用形态学方法进行鉴定;通过抗药性试验筛选出抗球虫效果较好的西药;根据鸡球虫生物学特性、鸡的生理特点,结合抗药性试验结果,在本实验室前期抗球虫中药复方研究的基础上,研制防治鸡球虫病的中西药复方制剂,并对中西药复方制剂抗球虫效果进行研究。
首先,从安徽省肥东县、肥西县、长丰县、五河县和宣城郊区等五个县(区)采集鸡的新鲜粪样,共采集172份。用饱和食盐水溶液漂浮法对每个粪样进行检查,球虫总感染率为100%。用沉浮法收集粪样中球虫卵囊,加入2.5%重铬酸钾水溶液,置29℃恒温培养箱中培养至孢子化。用所获孢子化卵囊进行人工鸡体增殖试验,对孢子化卵囊进行形态学观察,初步鉴定本次分离的7种球虫分别为毒害艾美耳球虫(Eimeria necatrix)、堆型艾美耳球虫(E.acervulina)、巨型艾美耳球虫(E.maxima)、柔嫩艾美耳球虫(E.tenella)、和缓艾美耳球虫(E.mitis)、哈氏艾美耳球虫(E.hagani)和早熟艾美耳球虫(E.praecox),均隶属于艾美耳属。
其次,选取地克珠利、尼卡巴嗪、托曲珠利、磺胺喹噁啉、磺胺氯吡嗪钠和盐霉素钠等6种药物进行鸡球虫抗药性试验,筛选出尼卡巴嗪和磺胺喹噁啉2种抗球虫效果较好的药物,为研制抗球虫中西药复方制剂提供了材料。
再次,分别将磺胺喹噁啉和尼卡巴嗪与马杜拉霉素、氨丙啉配伍,并进行鸡体试验,筛选出抗鸡球虫西药复方。将筛选出的西药复方与中药复方(本实验室提供)进行配制试验、剂量用药和时间选取鸡体试验,研制成中西药复方散剂,该制剂综合抗球虫指数(ACI)预防组达189、治疗组达187,其质量符合《中华人民共和国2000年药典》标准。
最后,进行了中西药复方制剂疗效试验。以混合孢子化球虫卵囊接种雏鸡,研究中西药复方制剂对球虫感染鸡血液生理生化指标的影响。结果表明,球虫感染后能引起鸡血清葡萄糖、镁、钠、总蛋白、白蛋白、琥珀酸脱氢酶、淀粉酶、碱性磷酸酶含量明显下降;血清钙、一氧化氮、溶菌酶、谷草转氨酶、谷丙转氨酶、IgG、IgM、IgA含量升高;血清超氧化物歧化酶、钾、尿素变化不明显。中西药复方制剂能够维持血糖等指标稳定在较正常水平,并能明显提高溶菌酶等指标的水平。中西药复方制剂能显著促进雏鸡生长,提高养鸡业的经济效益。
综上所述,本次对安徽部分地区鸡球虫感染情况进行了调查,其结果可为这些地区鸡球虫病的防治提供理论依据;研制出一个防治鸡球虫病的中西药复方制剂,所研制的中西药复方制剂不仅具有较好的抗球虫效果,而且能够促进雏鸡生长,提高饲料转化率。因此,本研究为鸡球虫病的防治提供了一个新选择,对提高养鸡业经济效益具有一定意义。
Chicken coccidiosis is caused by one kind or more kinds of Eimeria and happens worldwidely.The sickness causes harm to chicken seriously,which causes the huge economic loss in chickens industry.Chicken coccidiosis consists of a mixed infections caused by various coccidiosis,in which Eimeria tenella and E.necatrix's are stronger than others.This research separated Chicken coccidiosis from chicken's excrement in some areas of Anhui province.The identification of coccidiosis was carried by morphological method.Antibiotic experiment was carried on to find anti-coccidiosis medicine.According to the antibiotic nature test results,the Chicken coccidiosis biological characteristics and chicken's physiological characteristics,Chinese herb and antibiotic therapy method were develped on the basis of previous research of Chinese herb.The prevention effect on coccidiosis,and its effect on the disease were also studied.
Firstly,chicken's fresh excrement was gathered from folowing counties in Anhui province,namely,Feidong,Feixi,Changfeng,Wuhe and Xuancheng.Oogonium were collected by the up-and-down methed,suspended with 2.5%potassium solution and incubated at 29℃.And then mutipled in Chicken.Gathered oogonium were observed using light microsope,The results showed that the seprated ooganium consisted of Eimeria necatrix,E.acervulina,E.maxima,E.tenella,E.mitis,E.hagani and E.praecox.All of them were included in Eimeria.
Secondly,Sulfachloropyridazine pyrazine sodium,salinomycin,diclazuril, sulfaquinoxaline and Toltrazurila were tested for anti-coccidiosis effects.The results showed that the effect of Nicarbazin and sulfaquinoxaline were best.
Thirdly,after combining sullfaquinoxaline,Nicarbazin,Madubamycin and amprolium respectively,anti-coccidiosis prescription was selected in chicken.The research was to choose choice proper time,dose and prescription of chinese herb and anti-biotic medicine for preventing and treating coccidiosis.The results showed that anti-coccidiosis index were up to 189 and 187 in preventing group and curing group respectively.Its quality met the pharmacopoeia standard of People's Republic of China in 2000.
Fourthly,the experiment was carried out to prevent chiken coccidiosis using Chinese herb and western medicine compound prescription.We studied the change pattern of physiological biochemistry target in young chicken that was vaccinated after using Chinese herb and western medicine compound prescription.The results indicated that the content of GLU,Mg,Na,TP,ALB,SDH,AMS and ALP reduced significantly;and the content of Ca,NO,LZM,GOT and GPT increased,as well as IgG,IgM and IgA.But SOD,GU and K didn't change obviously.The level of GLU was maintained normally while LZM increased significantly.The Chinese and western medicine compound prescription increased obviously the chickling to grow and improved chickens industry and economic efficiency.
In summary,this study carried on the epidemiology investigation successfully in some areas of Anhui province.We develop a compound prescription of chicken coccidiosis's Chinese herb and western medicine,which not only has the good anti-coccidiosis effect,but also can promote the chickling to grow,improve the feed conversion rate.Therefore,this research has explored a new choice for the chicken coccidiosis's prevention.It can enhance economic effciency in chiken industy.
首先,从安徽省肥东县、肥西县、长丰县、五河县和宣城郊区等五个县(区)采集鸡的新鲜粪样,共采集172份。用饱和食盐水溶液漂浮法对每个粪样进行检查,球虫总感染率为100%。用沉浮法收集粪样中球虫卵囊,加入2.5%重铬酸钾水溶液,置29℃恒温培养箱中培养至孢子化。用所获孢子化卵囊进行人工鸡体增殖试验,对孢子化卵囊进行形态学观察,初步鉴定本次分离的7种球虫分别为毒害艾美耳球虫(Eimeria necatrix)、堆型艾美耳球虫(E.acervulina)、巨型艾美耳球虫(E.maxima)、柔嫩艾美耳球虫(E.tenella)、和缓艾美耳球虫(E.mitis)、哈氏艾美耳球虫(E.hagani)和早熟艾美耳球虫(E.praecox),均隶属于艾美耳属。
其次,选取地克珠利、尼卡巴嗪、托曲珠利、磺胺喹噁啉、磺胺氯吡嗪钠和盐霉素钠等6种药物进行鸡球虫抗药性试验,筛选出尼卡巴嗪和磺胺喹噁啉2种抗球虫效果较好的药物,为研制抗球虫中西药复方制剂提供了材料。
再次,分别将磺胺喹噁啉和尼卡巴嗪与马杜拉霉素、氨丙啉配伍,并进行鸡体试验,筛选出抗鸡球虫西药复方。将筛选出的西药复方与中药复方(本实验室提供)进行配制试验、剂量用药和时间选取鸡体试验,研制成中西药复方散剂,该制剂综合抗球虫指数(ACI)预防组达189、治疗组达187,其质量符合《中华人民共和国2000年药典》标准。
最后,进行了中西药复方制剂疗效试验。以混合孢子化球虫卵囊接种雏鸡,研究中西药复方制剂对球虫感染鸡血液生理生化指标的影响。结果表明,球虫感染后能引起鸡血清葡萄糖、镁、钠、总蛋白、白蛋白、琥珀酸脱氢酶、淀粉酶、碱性磷酸酶含量明显下降;血清钙、一氧化氮、溶菌酶、谷草转氨酶、谷丙转氨酶、IgG、IgM、IgA含量升高;血清超氧化物歧化酶、钾、尿素变化不明显。中西药复方制剂能够维持血糖等指标稳定在较正常水平,并能明显提高溶菌酶等指标的水平。中西药复方制剂能显著促进雏鸡生长,提高养鸡业的经济效益。
综上所述,本次对安徽部分地区鸡球虫感染情况进行了调查,其结果可为这些地区鸡球虫病的防治提供理论依据;研制出一个防治鸡球虫病的中西药复方制剂,所研制的中西药复方制剂不仅具有较好的抗球虫效果,而且能够促进雏鸡生长,提高饲料转化率。因此,本研究为鸡球虫病的防治提供了一个新选择,对提高养鸡业经济效益具有一定意义。
Chicken coccidiosis is caused by one kind or more kinds of Eimeria and happens worldwidely.The sickness causes harm to chicken seriously,which causes the huge economic loss in chickens industry.Chicken coccidiosis consists of a mixed infections caused by various coccidiosis,in which Eimeria tenella and E.necatrix's are stronger than others.This research separated Chicken coccidiosis from chicken's excrement in some areas of Anhui province.The identification of coccidiosis was carried by morphological method.Antibiotic experiment was carried on to find anti-coccidiosis medicine.According to the antibiotic nature test results,the Chicken coccidiosis biological characteristics and chicken's physiological characteristics,Chinese herb and antibiotic therapy method were develped on the basis of previous research of Chinese herb.The prevention effect on coccidiosis,and its effect on the disease were also studied.
Firstly,chicken's fresh excrement was gathered from folowing counties in Anhui province,namely,Feidong,Feixi,Changfeng,Wuhe and Xuancheng.Oogonium were collected by the up-and-down methed,suspended with 2.5%potassium solution and incubated at 29℃.And then mutipled in Chicken.Gathered oogonium were observed using light microsope,The results showed that the seprated ooganium consisted of Eimeria necatrix,E.acervulina,E.maxima,E.tenella,E.mitis,E.hagani and E.praecox.All of them were included in Eimeria.
Secondly,Sulfachloropyridazine pyrazine sodium,salinomycin,diclazuril, sulfaquinoxaline and Toltrazurila were tested for anti-coccidiosis effects.The results showed that the effect of Nicarbazin and sulfaquinoxaline were best.
Thirdly,after combining sullfaquinoxaline,Nicarbazin,Madubamycin and amprolium respectively,anti-coccidiosis prescription was selected in chicken.The research was to choose choice proper time,dose and prescription of chinese herb and anti-biotic medicine for preventing and treating coccidiosis.The results showed that anti-coccidiosis index were up to 189 and 187 in preventing group and curing group respectively.Its quality met the pharmacopoeia standard of People's Republic of China in 2000.
Fourthly,the experiment was carried out to prevent chiken coccidiosis using Chinese herb and western medicine compound prescription.We studied the change pattern of physiological biochemistry target in young chicken that was vaccinated after using Chinese herb and western medicine compound prescription.The results indicated that the content of GLU,Mg,Na,TP,ALB,SDH,AMS and ALP reduced significantly;and the content of Ca,NO,LZM,GOT and GPT increased,as well as IgG,IgM and IgA.But SOD,GU and K didn't change obviously.The level of GLU was maintained normally while LZM increased significantly.The Chinese and western medicine compound prescription increased obviously the chickling to grow and improved chickens industry and economic efficiency.
In summary,this study carried on the epidemiology investigation successfully in some areas of Anhui province.We develop a compound prescription of chicken coccidiosis's Chinese herb and western medicine,which not only has the good anti-coccidiosis effect,but also can promote the chickling to grow,improve the feed conversion rate.Therefore,this research has explored a new choice for the chicken coccidiosis's prevention.It can enhance economic effciency in chiken industy.
引文
[1]孔繁瑶.家畜寄生虫学[M].北京:中国农业大学出版社,1997:328-334.
[2]汪明.兽医寄生虫学[M].北京:中国农业出版社,2003:284-288.
[3]彼得L朗.球虫生物学[M].蒋金书,刘钟灵,陆信武,等译.广西:广西科学技术出版社,1990:103-120.
[4]Fabrice Laurent,Roselyne Mancassola,Sonla Lacroix et al.Analysis of Chicken Mucosal Immune Response to Eimeria tenella and Eimeria maxima Infection by Quantitative Reverse Transcription-PCR[J].Infection and Immuity,2001,69(4):2527-2534.
[5]索勋,李国清.鸡球虫病学[M].北京:中国农业大学出版社,1998:238-268.
[6]缪德年,樊生超,姚惠娟,等.鸡球虫虫种分离鉴别方法研究[J].国外畜牧学-猪与禽,1999,1:51-52.
[7]北京农业大学主编.家畜寄生虫学[M].北京:农业出版社,1981:416-420.
[8]蒋金书.动物原虫病学[M].北京:中国农业大学出版社,2000
[9]S.H.M.Jeurissen.Eimeria tenella infections in chickens:aspects of host parasite:interaction[J].Veterinary Immunology and Immunopathology,1996,54(2):231-238
[10]安健,汪明,孔繁瑶等.柔嫩艾美耳球虫大配子发育和卵囊的超微结构[J].中国兽医学报,2001,21(6):559-561
[11]安健,汪明,孔繁瑶等.柔嫩艾美耳球虫裂殖生殖阶段超微结构的研究[J].畜牧兽医学报,2001,32(5):452-457
[12]安健,汪明,孔繁瑶等.柔嫩艾美耳球虫配子发生的超微结构[J].动物学报,2001,47(4):431-435
[13]林青.鸡球虫单卵囊分离技术的构建及YL地理株的致病性与抗药性研究[硕士学位论文].杨陵:西北农林科技大学,2002.
[14]缪德年,樊生超,姚惠娟,等.鸡球虫虫种分离鉴别方法研究[J].国外畜牧学-猪与禽,1999,1·51-52.
[15]Levine N D.Veterinary Protozoology[M].Ames,Iowa:the Iowa State University Press,1985:183-184.
[16]卡尔尼克B W.禽病学[M].第9版.高福,刘文军主译.北京:北京农业大学出版社,1991:682-692.
[17]陶建平,辛玲,李文超,等.巨型艾美耳球虫(Eimeda maxima)地理株研究Ⅰ.11株巨型艾美耳球虫的分离与基本特征[J].中国兽医学报,2004,24(4):349-352.
[18]聂奎,胡世君,汤明.重庆市鸡球虫种类调查[J].黑龙江畜牧兽医,2002,(11):24-25.
[19]曾燕玲,赵福葳,陈能桥.遵义市鸡球虫种类的调查[J].中国兽医寄生虫病,2004,12(3):22-23.
[20]杨发龙,蒋锡仕,李必富,等.成都地区鸡球虫种类的初步调查[J].四川畜牧兽医,2001,(4):28-29.
[21]杜爱芳,李肖梁,王素华.杭州地区鸡球虫种类研究[J].浙江农业学报,2002,14(2):100-104.
[22]陈丽凤,李蕴玉,张香斋,等.秦皇岛地区十个鸡场鸡球虫种类的初步调查[J].中国兽医寄生虫病,2003,11(1):37-39.
[23]李培英,彭克森,张学道.安徽合肥市鸡球虫种类及感染情况调查[J].中国兽医寄生虫病,2004,12(3):24-26.
[24]苑淑贤,姚新华,任科研,等.柔嫩艾美耳球虫纯种的分离鉴定和致病性观察[J].农业与技术,2005,25(4):90-92.
[25]张龙现,蒋金书,刘群,等.毒害艾美耳球虫纯种卵囊收集鉴定及致病性测定[J].中国兽医杂志,2001,37(9):12-13.
[26]安健,汪明,韩春来,等.柔嫩艾美球虫裂殖子的分离纯化[J].中国兽医科技,2003,33(12):54-57.
[27]黄兵,吴薛忠,史天卫,等.毒害艾美球虫纯种分离的初步鉴定和致病性研究[J].中国兽医寄生虫病,1993,1(4):24-28.
[28]李凯,周本江,杨照青.球虫卵囊的分子生物学研究进展[J].中国寄生虫学与寄生虫病杂志,2005.23(3):178-181.
[29]Gasser RB.PCR-based technology in veterinary parasitology[J].Vet.Parasitol,1999,84:229-258.
[30]W.G.Woods,K.G.Whithear,D.G.Richards et al.Single-strand restriction fragment length polymorphism analysis of the second internal transcribed spacer(ribosomal DNA) for six species of Eimeria from chickens in Australia[J].International Journal for Parasitology,2000,30:1019-1023
[31]缪德年,樊生超,姚惠娟等.鸡球虫虫种分离鉴别方法研究[J].国外畜牧学-猪与禽,1999,1:51-52
[32]Guay.J-M.Dubois.D.Morency,Gagon,S et al.Detection of the pathogenic parasite Toxoplasma gondii by specific amplification of ribosomal sequences using comultiples polymerase chain reaction[J].Jounal of Clinical Microbiology,1993,31:203-207
[33]Holmdahl.O.J.M.& Mattsson.J.G.Rapid and sensitive identification of Neospora caninum by in viro amplification of the inemal transcribed spacer 1[J].Parasitology,1996,112:177-182
[34]Schnitzler BE,Thebo PL,Mattsson JG et al.Development of a diagnostic PCR assay for the detection and discrimination of four pathogenic Eimeria species of the chicken[J].Avian Pathology,1998,27:490-497
[35]Schnitzler BE,Thebo PL,Tomley FM et al.PCR identification of chicken Eimeria:a simplified read-out[J].Avian Pathology,1999,28:89-93
[36]Yao-Chi Su,Andrew Chang-Young Fei.Differential diagnosis of five avian Eimeria species by polymerase chain reaction using primers derived from the internal transcribed spacer 1(ITS-1)sequence[J].Veterinary Parasitology,2003,117:221-227
[37]Waitumbi J N,Murphy N B.Infra-and infra-species differentiation of trypanosomes by genomic fingerprinting with arbitrary primers[J].Molicular and Biochemical Parasitology,1993,58:181-186
[38]Awad F M.Detection and species identification of Cryptosporidium oocysts using a system based on PCR and endonuclease restrction[J].Parasitol,1994,109:19-22
[39]MacPherson J M,Grajadhar A A.Sensitive and specific polymerase chain reaction detection of Toxoplasima gondii for veterinary and medical diagnosis[J].Can J Vet Res,1992,57:45-48
[40]MacPherson J M,Gajadhar A A.Specific amplification of Sorcocystis cruzi DNA using a randomly primed polymerase chain reaction assay[J].Vet Parasitology,1994,55:267-277
[41]Macpherson J M,Gajadhar A A.Differentiation of seven Eimeria species by random amplified polymorphic DNA[J].Vet Parasitology,1993,45:257-266
[42]Macpherson J M,Gajadhar A A.Differentiation of seven Eimeria species by random amplified polymorphic DNA[J].Vet Parasitology,1993,45:257-266
[43]Fernando MA,Pastemak JJ.Eimeria spp.of the domestic fowl:resolution of chromosomes by field inversion gel electrophoresis[J].Exp Parasitol.1991,72(3):306-310.
[44]陈仗榴.兽医药理学[M].中国农业出版社.2002.1
[45]MaesL,Coussement W,Vanparijs,et al.[J]JParasitology,1988,74(6):931-938
[46]赵其平,吴薛忠.马杜拉霉素与地克珠利联合用药预防鸡E.tenella球虫笼饲试验[J].中国兽医寄生虫病,1999,7(3):12-13
[47]聂奎,颜焰,高俊伟,等.地克珠利、氨丙啉与左旋咪唑联合使用对E.tenella的效力试验[J].中国兽医寄生虫病,2006,1(14):1-4
[48]缪德年,樊生超,录慧娟.鸡球虫病防治策略[J].中国禽业导刊,1998,14(2):48-49
[49]史晓丽,余祖功,江善祥.马杜拉霉素联合尼卡巴嗪的抗球虫效果与毒性试验[J].畜牧与兽医,1997,29(6):246-247
[50]许金俊,陶建平,彭金彪等.柔嫩艾美耳球虫扬州分离株对8种抗球虫药的抗药性[J]畜牧与兽医,2008,40(1):18-21
[51]Du A,Hu S.Effects of a Herbal Complex Against Eimeria tenella Infection in Chickens[J].J Vet Med B Infect Dis Vet Public Health,2004,51(4):194-197
[52]陈汉忠,李桂庆,李致宝等.中药与化学药物对鸡球虫病的疗效对比试验[J].中国家禽学报,2004,1(8):23-24
[53]商云霞,腾德全,王培.不同种类抗球虫药对常见鸡球虫效果的评价试验[J].甘肃畜牧兽医,2004(3):2-4
[54]董发明,王天奇,龙塔.等中西复方制剂“球霸”对人工感染鸡球虫病疗效试验[J].中国兽医杂志2005,41(2):32-33
[55]东彦新,李景峰中草药预防雏鸡堆型艾美耳球虫病试验[J]中国家禽,2008,30(6):46-48
[56]缪德年,姚惠娟,樊生超.中西药结合预防鸡球虫病试验[J].畜牧与兽医,2002 9(6):26-27
[57]李致宝.化学抗球虫药和中草药对球虫病的疗效对比试验[J].畜禽业,2000(8):16-18
[58]于咏兰,汪明.一种复方抗球虫制剂的研制[J].中国兽医杂志,2006,42(9):49-50
[59]颜卉,郗伟斌,杜绍范.中西结合制剂防治鸡柔嫩艾美耳球虫病试验[J].中国兽医杂志,2006,42(5):37-38
[60]孔繁瑶,宁长申,殷佩云,等.15株柔嫩艾美耳球虫Eimeria tenella对5种抗球虫药的抗药性调查[J].北京农业大学学报,1994,20(3):302-307.
[61]汪明,孔繁瑶,殷佩云,等.10株柔嫩艾美耳球虫对四种药物的抗药性检测[J].中国农业大学学报,1996,1(5):110-114.
[62]曾明华,胡志平.柔嫩艾美耳球虫野外株对3种聚醚类离子载体抗生素的敏感性试验[J].中国兽医学报,1996,16(4):390-393.
[63]Elizabeth Culotta.Funding cruch hobbles antibiotic resistance research[J].Science,1994,264(15):362-363.
[64]白喜婷.中西药合用防治鸡球虫病试验.河南科技大学学报[J],2006,24(1):17-19
[65]李佩国,李蕴玉,刘建辉等中西药复方抗鸡球虫病的效果观察.中兽医医药杂志[J],2002,2:12-14
[66]缪德年,姚惠娟,樊生超.中西药结合防治鸡球虫病的试验,畜牧与兽医[J],2002,34(6):26-27
[67]葛凯,李培英.中草药防治鸡球虫病研究进展[J].动物医学进展,2006,27(2):17-21.
[68]颜卉,郗伟斌,杜绍范.中药复方制剂抗鸡球虫效果的研究[J].黑龙江畜牧兽医,2005,(6):68-69.
[69]王天奇,范文利,董发明,等.中药煎剂抗鸡柔嫩艾美耳球虫的效果研究[J].中兽医医药杂志,2005,(2):29-31.
[70]金光明,丁希强.直杀球虫与强力灭球王对鸡柔嫩艾美耳球虫的疗效观察[J].中国畜牧兽 医,2004,31(12):42-44.
[71]景诗韵,谢百炼,吴传刚,等.中药对鸡人工感染球虫病免疫功能的影响[J].贵州畜牧兽医,2001,25(1):1-2.
[72]臧海军,张克英.动物肠道正常菌群对粘膜免疫影响的研究进展[J].饲料博览,2007,(3):35-37.
[73]郭莉,顾小龙,秦建华.复方中草药免疫增强剂对鸡堆型艾美耳球虫早熟株免疫的影响研究[J].中同家禽,2006,28(11):21-23.
[74]韦进钟,吴玄光,林辉环.感染堆形艾美球虫的鸡血清生化指标检测[J].中国兽医科技,2002,32(8):25-27.
[75]连宏军,丁熙成,濮红.雏鸡巨型艾美耳球虫感染后对血液若干生理指标的影响[J].中国兽医杂志,2000,26(5):23-24.
[76]李玉焕.家禽球虫感染的生理生化变化[J].中国兽医寄生虫病,1996,4(2):59-60.
[77]陈刚,蒋金书,蒋进.鸡感染巨型艾美耳球虫后血清生化指标的变化[J].中国兽医杂志,1999,25(7):6-8.
[78]杜爱芳.药物对球虫感染鸡血液及生化指标的影响[J].中国兽医学报,2002,22(1):73-75.
[79]顾有方,罗浩,陈会良.柔嫩艾美耳球虫感染对鸡的若干生理生化指标的影响[J].畜牧与兽医,2003,35(5):13-15.
[80]Allen P C.Production of free radical species during Eimeria maxima infections in chickens[J].Poultry Science,1997,76:814-821.
[81]包永占,史万玉,秦建华,等.不同剂量柔嫩艾美耳球虫感染对雏鸡脾重与血清NO含量的影响[J].中国兽医杂志,2005,41(10):20-21.
[82]Bhogal B S,Miller G A.Potential of a recombinant antigen as a prophylactic vaccine for one-day-old broiler chickens against E.acervulina and E.tenella infections[J].Veterinary Immunology and Immunopathology,1992,31:323-335.
[83]李祥瑞.球虫感染与家禽生理生化变化[J].畜牧与兽医,1990,22(3):137-139.
[84]Witlock D R.Physio logical basis of Eimeria tenella induced mortality in individual chickens[J].J Parasitol,1981,67(1):65-69.
[85]王晓良.钾离子通道在神经元凋亡中的重要作用药理通讯[J],2005,23(5)4-5
[86]李彩燕,黄春萍,于增华等.酸血症和痛风[J]中华临床医药2004(5)8:45-47
[87]卓曲.几种中草药抗鸡球虫的研究[硕士学位论文].广州:华南农业大学,2002.
[88]刘晶,郑世民,胡京友.雏鸡感染毒害艾美耳球虫后的体液免疫[J].中国兽医学报,2006,26(4):373-375.
[89]陈缓,周玫.自由基医学[M].北京:人民出版社,1991:122-140.
[90]Allen,P.C.Nitric Oxide Production During Eimeria tenella Infections in Chickens[J].Poultry Science,1997,76:810-813.
[91]韦进钟,吴玄光,林辉环.感染柔嫩艾美耳球虫后某些血清生化指标变化的研究[J].中国兽医杂志,2003,39(3):8-11.
[92]韦进钟,吴玄光,林辉环.感染堆型艾美耳球虫的鸡血清生化指标检测[J].中国兽医科技,2002,32(8):25-27
[93]李佩国,贾清辉,李蕴玉等.中药对球虫感染雏鸡血清乳酸脱氢酶和碱性磷酸酶活性的影响[J].河北科技师范学院学报,2007,21(1):1-3
[94]朱忠勇.实用医学检验学[M].北京:人民军医出版社,1992:95-115.
[95]李敏.溶茵酶及其应用[J].生物学教学,2006,31(4):2-3.
[2]汪明.兽医寄生虫学[M].北京:中国农业出版社,2003:284-288.
[3]彼得L朗.球虫生物学[M].蒋金书,刘钟灵,陆信武,等译.广西:广西科学技术出版社,1990:103-120.
[4]Fabrice Laurent,Roselyne Mancassola,Sonla Lacroix et al.Analysis of Chicken Mucosal Immune Response to Eimeria tenella and Eimeria maxima Infection by Quantitative Reverse Transcription-PCR[J].Infection and Immuity,2001,69(4):2527-2534.
[5]索勋,李国清.鸡球虫病学[M].北京:中国农业大学出版社,1998:238-268.
[6]缪德年,樊生超,姚惠娟,等.鸡球虫虫种分离鉴别方法研究[J].国外畜牧学-猪与禽,1999,1:51-52.
[7]北京农业大学主编.家畜寄生虫学[M].北京:农业出版社,1981:416-420.
[8]蒋金书.动物原虫病学[M].北京:中国农业大学出版社,2000
[9]S.H.M.Jeurissen.Eimeria tenella infections in chickens:aspects of host parasite:interaction[J].Veterinary Immunology and Immunopathology,1996,54(2):231-238
[10]安健,汪明,孔繁瑶等.柔嫩艾美耳球虫大配子发育和卵囊的超微结构[J].中国兽医学报,2001,21(6):559-561
[11]安健,汪明,孔繁瑶等.柔嫩艾美耳球虫裂殖生殖阶段超微结构的研究[J].畜牧兽医学报,2001,32(5):452-457
[12]安健,汪明,孔繁瑶等.柔嫩艾美耳球虫配子发生的超微结构[J].动物学报,2001,47(4):431-435
[13]林青.鸡球虫单卵囊分离技术的构建及YL地理株的致病性与抗药性研究[硕士学位论文].杨陵:西北农林科技大学,2002.
[14]缪德年,樊生超,姚惠娟,等.鸡球虫虫种分离鉴别方法研究[J].国外畜牧学-猪与禽,1999,1·51-52.
[15]Levine N D.Veterinary Protozoology[M].Ames,Iowa:the Iowa State University Press,1985:183-184.
[16]卡尔尼克B W.禽病学[M].第9版.高福,刘文军主译.北京:北京农业大学出版社,1991:682-692.
[17]陶建平,辛玲,李文超,等.巨型艾美耳球虫(Eimeda maxima)地理株研究Ⅰ.11株巨型艾美耳球虫的分离与基本特征[J].中国兽医学报,2004,24(4):349-352.
[18]聂奎,胡世君,汤明.重庆市鸡球虫种类调查[J].黑龙江畜牧兽医,2002,(11):24-25.
[19]曾燕玲,赵福葳,陈能桥.遵义市鸡球虫种类的调查[J].中国兽医寄生虫病,2004,12(3):22-23.
[20]杨发龙,蒋锡仕,李必富,等.成都地区鸡球虫种类的初步调查[J].四川畜牧兽医,2001,(4):28-29.
[21]杜爱芳,李肖梁,王素华.杭州地区鸡球虫种类研究[J].浙江农业学报,2002,14(2):100-104.
[22]陈丽凤,李蕴玉,张香斋,等.秦皇岛地区十个鸡场鸡球虫种类的初步调查[J].中国兽医寄生虫病,2003,11(1):37-39.
[23]李培英,彭克森,张学道.安徽合肥市鸡球虫种类及感染情况调查[J].中国兽医寄生虫病,2004,12(3):24-26.
[24]苑淑贤,姚新华,任科研,等.柔嫩艾美耳球虫纯种的分离鉴定和致病性观察[J].农业与技术,2005,25(4):90-92.
[25]张龙现,蒋金书,刘群,等.毒害艾美耳球虫纯种卵囊收集鉴定及致病性测定[J].中国兽医杂志,2001,37(9):12-13.
[26]安健,汪明,韩春来,等.柔嫩艾美球虫裂殖子的分离纯化[J].中国兽医科技,2003,33(12):54-57.
[27]黄兵,吴薛忠,史天卫,等.毒害艾美球虫纯种分离的初步鉴定和致病性研究[J].中国兽医寄生虫病,1993,1(4):24-28.
[28]李凯,周本江,杨照青.球虫卵囊的分子生物学研究进展[J].中国寄生虫学与寄生虫病杂志,2005.23(3):178-181.
[29]Gasser RB.PCR-based technology in veterinary parasitology[J].Vet.Parasitol,1999,84:229-258.
[30]W.G.Woods,K.G.Whithear,D.G.Richards et al.Single-strand restriction fragment length polymorphism analysis of the second internal transcribed spacer(ribosomal DNA) for six species of Eimeria from chickens in Australia[J].International Journal for Parasitology,2000,30:1019-1023
[31]缪德年,樊生超,姚惠娟等.鸡球虫虫种分离鉴别方法研究[J].国外畜牧学-猪与禽,1999,1:51-52
[32]Guay.J-M.Dubois.D.Morency,Gagon,S et al.Detection of the pathogenic parasite Toxoplasma gondii by specific amplification of ribosomal sequences using comultiples polymerase chain reaction[J].Jounal of Clinical Microbiology,1993,31:203-207
[33]Holmdahl.O.J.M.& Mattsson.J.G.Rapid and sensitive identification of Neospora caninum by in viro amplification of the inemal transcribed spacer 1[J].Parasitology,1996,112:177-182
[34]Schnitzler BE,Thebo PL,Mattsson JG et al.Development of a diagnostic PCR assay for the detection and discrimination of four pathogenic Eimeria species of the chicken[J].Avian Pathology,1998,27:490-497
[35]Schnitzler BE,Thebo PL,Tomley FM et al.PCR identification of chicken Eimeria:a simplified read-out[J].Avian Pathology,1999,28:89-93
[36]Yao-Chi Su,Andrew Chang-Young Fei.Differential diagnosis of five avian Eimeria species by polymerase chain reaction using primers derived from the internal transcribed spacer 1(ITS-1)sequence[J].Veterinary Parasitology,2003,117:221-227
[37]Waitumbi J N,Murphy N B.Infra-and infra-species differentiation of trypanosomes by genomic fingerprinting with arbitrary primers[J].Molicular and Biochemical Parasitology,1993,58:181-186
[38]Awad F M.Detection and species identification of Cryptosporidium oocysts using a system based on PCR and endonuclease restrction[J].Parasitol,1994,109:19-22
[39]MacPherson J M,Grajadhar A A.Sensitive and specific polymerase chain reaction detection of Toxoplasima gondii for veterinary and medical diagnosis[J].Can J Vet Res,1992,57:45-48
[40]MacPherson J M,Gajadhar A A.Specific amplification of Sorcocystis cruzi DNA using a randomly primed polymerase chain reaction assay[J].Vet Parasitology,1994,55:267-277
[41]Macpherson J M,Gajadhar A A.Differentiation of seven Eimeria species by random amplified polymorphic DNA[J].Vet Parasitology,1993,45:257-266
[42]Macpherson J M,Gajadhar A A.Differentiation of seven Eimeria species by random amplified polymorphic DNA[J].Vet Parasitology,1993,45:257-266
[43]Fernando MA,Pastemak JJ.Eimeria spp.of the domestic fowl:resolution of chromosomes by field inversion gel electrophoresis[J].Exp Parasitol.1991,72(3):306-310.
[44]陈仗榴.兽医药理学[M].中国农业出版社.2002.1
[45]MaesL,Coussement W,Vanparijs,et al.[J]JParasitology,1988,74(6):931-938
[46]赵其平,吴薛忠.马杜拉霉素与地克珠利联合用药预防鸡E.tenella球虫笼饲试验[J].中国兽医寄生虫病,1999,7(3):12-13
[47]聂奎,颜焰,高俊伟,等.地克珠利、氨丙啉与左旋咪唑联合使用对E.tenella的效力试验[J].中国兽医寄生虫病,2006,1(14):1-4
[48]缪德年,樊生超,录慧娟.鸡球虫病防治策略[J].中国禽业导刊,1998,14(2):48-49
[49]史晓丽,余祖功,江善祥.马杜拉霉素联合尼卡巴嗪的抗球虫效果与毒性试验[J].畜牧与兽医,1997,29(6):246-247
[50]许金俊,陶建平,彭金彪等.柔嫩艾美耳球虫扬州分离株对8种抗球虫药的抗药性[J]畜牧与兽医,2008,40(1):18-21
[51]Du A,Hu S.Effects of a Herbal Complex Against Eimeria tenella Infection in Chickens[J].J Vet Med B Infect Dis Vet Public Health,2004,51(4):194-197
[52]陈汉忠,李桂庆,李致宝等.中药与化学药物对鸡球虫病的疗效对比试验[J].中国家禽学报,2004,1(8):23-24
[53]商云霞,腾德全,王培.不同种类抗球虫药对常见鸡球虫效果的评价试验[J].甘肃畜牧兽医,2004(3):2-4
[54]董发明,王天奇,龙塔.等中西复方制剂“球霸”对人工感染鸡球虫病疗效试验[J].中国兽医杂志2005,41(2):32-33
[55]东彦新,李景峰中草药预防雏鸡堆型艾美耳球虫病试验[J]中国家禽,2008,30(6):46-48
[56]缪德年,姚惠娟,樊生超.中西药结合预防鸡球虫病试验[J].畜牧与兽医,2002 9(6):26-27
[57]李致宝.化学抗球虫药和中草药对球虫病的疗效对比试验[J].畜禽业,2000(8):16-18
[58]于咏兰,汪明.一种复方抗球虫制剂的研制[J].中国兽医杂志,2006,42(9):49-50
[59]颜卉,郗伟斌,杜绍范.中西结合制剂防治鸡柔嫩艾美耳球虫病试验[J].中国兽医杂志,2006,42(5):37-38
[60]孔繁瑶,宁长申,殷佩云,等.15株柔嫩艾美耳球虫Eimeria tenella对5种抗球虫药的抗药性调查[J].北京农业大学学报,1994,20(3):302-307.
[61]汪明,孔繁瑶,殷佩云,等.10株柔嫩艾美耳球虫对四种药物的抗药性检测[J].中国农业大学学报,1996,1(5):110-114.
[62]曾明华,胡志平.柔嫩艾美耳球虫野外株对3种聚醚类离子载体抗生素的敏感性试验[J].中国兽医学报,1996,16(4):390-393.
[63]Elizabeth Culotta.Funding cruch hobbles antibiotic resistance research[J].Science,1994,264(15):362-363.
[64]白喜婷.中西药合用防治鸡球虫病试验.河南科技大学学报[J],2006,24(1):17-19
[65]李佩国,李蕴玉,刘建辉等中西药复方抗鸡球虫病的效果观察.中兽医医药杂志[J],2002,2:12-14
[66]缪德年,姚惠娟,樊生超.中西药结合防治鸡球虫病的试验,畜牧与兽医[J],2002,34(6):26-27
[67]葛凯,李培英.中草药防治鸡球虫病研究进展[J].动物医学进展,2006,27(2):17-21.
[68]颜卉,郗伟斌,杜绍范.中药复方制剂抗鸡球虫效果的研究[J].黑龙江畜牧兽医,2005,(6):68-69.
[69]王天奇,范文利,董发明,等.中药煎剂抗鸡柔嫩艾美耳球虫的效果研究[J].中兽医医药杂志,2005,(2):29-31.
[70]金光明,丁希强.直杀球虫与强力灭球王对鸡柔嫩艾美耳球虫的疗效观察[J].中国畜牧兽 医,2004,31(12):42-44.
[71]景诗韵,谢百炼,吴传刚,等.中药对鸡人工感染球虫病免疫功能的影响[J].贵州畜牧兽医,2001,25(1):1-2.
[72]臧海军,张克英.动物肠道正常菌群对粘膜免疫影响的研究进展[J].饲料博览,2007,(3):35-37.
[73]郭莉,顾小龙,秦建华.复方中草药免疫增强剂对鸡堆型艾美耳球虫早熟株免疫的影响研究[J].中同家禽,2006,28(11):21-23.
[74]韦进钟,吴玄光,林辉环.感染堆形艾美球虫的鸡血清生化指标检测[J].中国兽医科技,2002,32(8):25-27.
[75]连宏军,丁熙成,濮红.雏鸡巨型艾美耳球虫感染后对血液若干生理指标的影响[J].中国兽医杂志,2000,26(5):23-24.
[76]李玉焕.家禽球虫感染的生理生化变化[J].中国兽医寄生虫病,1996,4(2):59-60.
[77]陈刚,蒋金书,蒋进.鸡感染巨型艾美耳球虫后血清生化指标的变化[J].中国兽医杂志,1999,25(7):6-8.
[78]杜爱芳.药物对球虫感染鸡血液及生化指标的影响[J].中国兽医学报,2002,22(1):73-75.
[79]顾有方,罗浩,陈会良.柔嫩艾美耳球虫感染对鸡的若干生理生化指标的影响[J].畜牧与兽医,2003,35(5):13-15.
[80]Allen P C.Production of free radical species during Eimeria maxima infections in chickens[J].Poultry Science,1997,76:814-821.
[81]包永占,史万玉,秦建华,等.不同剂量柔嫩艾美耳球虫感染对雏鸡脾重与血清NO含量的影响[J].中国兽医杂志,2005,41(10):20-21.
[82]Bhogal B S,Miller G A.Potential of a recombinant antigen as a prophylactic vaccine for one-day-old broiler chickens against E.acervulina and E.tenella infections[J].Veterinary Immunology and Immunopathology,1992,31:323-335.
[83]李祥瑞.球虫感染与家禽生理生化变化[J].畜牧与兽医,1990,22(3):137-139.
[84]Witlock D R.Physio logical basis of Eimeria tenella induced mortality in individual chickens[J].J Parasitol,1981,67(1):65-69.
[85]王晓良.钾离子通道在神经元凋亡中的重要作用药理通讯[J],2005,23(5)4-5
[86]李彩燕,黄春萍,于增华等.酸血症和痛风[J]中华临床医药2004(5)8:45-47
[87]卓曲.几种中草药抗鸡球虫的研究[硕士学位论文].广州:华南农业大学,2002.
[88]刘晶,郑世民,胡京友.雏鸡感染毒害艾美耳球虫后的体液免疫[J].中国兽医学报,2006,26(4):373-375.
[89]陈缓,周玫.自由基医学[M].北京:人民出版社,1991:122-140.
[90]Allen,P.C.Nitric Oxide Production During Eimeria tenella Infections in Chickens[J].Poultry Science,1997,76:810-813.
[91]韦进钟,吴玄光,林辉环.感染柔嫩艾美耳球虫后某些血清生化指标变化的研究[J].中国兽医杂志,2003,39(3):8-11.
[92]韦进钟,吴玄光,林辉环.感染堆型艾美耳球虫的鸡血清生化指标检测[J].中国兽医科技,2002,32(8):25-27
[93]李佩国,贾清辉,李蕴玉等.中药对球虫感染雏鸡血清乳酸脱氢酶和碱性磷酸酶活性的影响[J].河北科技师范学院学报,2007,21(1):1-3
[94]朱忠勇.实用医学检验学[M].北京:人民军医出版社,1992:95-115.
[95]李敏.溶茵酶及其应用[J].生物学教学,2006,31(4):2-3.