宁夏部分地区奶牛球虫感染情况调查与遗传进化分析

doi:10.3969/j.issn.1004-1524.2021.08.04

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (8): 1379-1384.DOI: 10.3969/j.issn.1004-1524.2021.08.04

宁夏部分地区奶牛球虫感染情况调查与遗传进化分析

赵洪喜(), 刘继兵

宁夏大学农学院,宁夏银川 750021

收稿日期:2020-10-14 出版日期:2021-08-25 发布日期:2021-08-27
作者简介:赵洪喜(1977—),男,内蒙古赤峰人,博士,副教授,研究方向为动物寄生虫及其分子生物学。E-mail: zhaohongxi2006@163.com
基金资助:
国家自然科学基金(31760727);宁夏自然科学基金(NZ17007)

Infection investigation and genetic evolution analysis of dairy cow coccidiosis in parts of Ningxia

ZHAO Hongxi(), LIU Jibing

Colloge of Agriculture, Ningxia University, Yinchuan 750021, China

Received:2020-10-14 Online:2021-08-25 Published:2021-08-27

摘要/Abstract

摘要：

为了解宁夏地区奶牛球虫的感染现状与流行特征,以宁夏吴忠、石嘴山、贺兰地区5个不同规模化奶牛场采集的犊牛腹泻粪便样品为材料,利用饱和NaCl溶液飘浮法和麦克马斯特虫卵计数法观察统计球虫形态、种类与感染情况;并提取球虫卵囊基因组DNA,进行18S rRNA基因的扩增和测序,用DNAstar软件对奶牛球虫18S rRNA基因进行序列分析,并构建系统发育树。结果显示,在随机抽查的179份样品中,牛球虫病的平均感染率为56.98%,感染率的范围为40.00%~80.84%,平均每g粪便虫卵数(OPG)为1 358。孢子化卵囊呈圆形或椭圆形,初步确定该地区优势虫种为邱氏艾美尔球虫、牛艾美尔球虫,其大小为(25.7~36.0)μm×(21.4~27.1)μm。序列分析表明,宁夏地区所分离的虫株与其他虫株亲缘关系较远,但本地虫株之间有较近的亲缘关系。该研究丰富了奶牛球虫流行病学材料,也为该病的防治研究奠定了基础。

关键词: 宁夏, 奶牛, 球虫, 遗传进化

Abstract:

To investigate dairy cow coccidiosis infection status and epidemic characteristics in Ningxia, the fecal samples of calves with diarrhea collected from five large scale dairy farm in Wuzhong, Shizuishan and Helan of Ningxia were collected. The morphology, species and infection status of pathogeny were counted by saturated NaCl floating method and McMaster Egg Slide Counting method. Genomic DNA of coccidial oocyst was extracted for amplification and sequencing of 18S rRNA. DNAstar was used for sequence analysis with the published 18S rRNA gene of coccidia of dairy cattle, and a phylogenetic tree was constructed. Results showed that in the random of 179 samples, the average cow coccidiosis infection rate was 56.98%, the range of infection rate was 40.00%-80.84%, and average number of oocysts per gram feces (OPG) value was 1 358. Morphological observation showed that sporogenous oocysts were round or elliptic, and preliminarily demonstrated the dominant species in this area were Eimeria zuerni and Eimeria bovis, whose size were (25.7-36.0)μm×(21.4-27.1)μm. Sequence analysis showed that strains isolated from Ningxia had a distant relationship with other strains, but there was a close relationship between the local strains. This study not only enriched the epidemiological materials of dairy cow coccidiosis, but also laid a foundation for the prevention and treatment of the disease.

Key words: Ningxia, cow, coccidia, genetic evolution

中图分类号:

S858.23

赵洪喜, 刘继兵. 宁夏部分地区奶牛球虫感染情况调查与遗传进化分析[J]. 浙江农业学报, 2021, 33(8): 1379-1384.

ZHAO Hongxi, LIU Jibing. Infection investigation and genetic evolution analysis of dairy cow coccidiosis in parts of Ningxia[J]. Acta Agriculturae Zhejiangensis, 2021, 33(8): 1379-1384.

图/表 4

表1 奶牛场球虫感染率与OPG值

Table 1 Infection rate and OPG of coccidia in dairy farms

牧场 Farms	被检数量 Number examined	感染头数 Number infected	感染率 Infection rate/%	OPG值OPG value
牧场 Farms	被检数量 Number examined	感染头数 Number infected	感染率 Infection rate/%	范围Range	平均Average
1	30	15	50.00	100~1 800	840
2	44	35	80.84	100~4 800	2 752
3	32	18	50.50	100~2 400	1 624
4	38	20	55.56	100~1 200	823
5	35	14	40.00	100~3 600	754
总计Total	179	99	56.98	100~4 800	1 358

图1 球虫孢子化卵囊显微镜观察图 A,邱氏艾美尔球虫;B,柱状艾美尔球虫; C,牛艾美尔球虫; D,加拿大艾美尔球虫;E,皮利他艾美耳球虫;F,椭圆艾美尔球虫。

Fig.1 Observations of sporulated coccidia oocysts by microscope A, Eimeria zurnii; B, Eimeria cylindriac; C, Eimeria bovis; D, Eimeria canadensis; E, Eimeria pellita; F, Eimeria ellipsoidalis.

图2 奶牛球虫18S rRNA PCR扩增产物 M,DL2000 DNA marker;1~8:奶牛球虫扩增产物。

Fig.2 PCR amplification of 18S rRNA from dairy coccidia M, DL2000 DNA marker; 1-8, Amplified product of dairy coccidian.

图3 奶牛球虫18S rRNA基因序列的进化分析

Fig.3 Phylogenetic analysis of bovine coccidia based on 18S rRNA gene sequence

参考文献 22

[1]	孔繁瑶. 家畜寄生虫学:第二版[M]. 北京: 中国农业大学出版社, 1997: 341-343.
[2]	张龙现, 宁长申, 何青军, 等. 郑州奶牛球虫病流行病学调查与虫种鉴定[J]. 河南农业大学学报, 2000, 34(1):88-93.
	ZHANG L X, NING C S, HE Q J, et al. Survey of epidemiology of dairy cow coccidiosis in Zhengzhou and identification of Coccidia species[J]. Journal of Henan Agricultural University, 2000, 34(1):88-93.(in Chinese with English abstract)
[3]	ASFAW D, DENEKE Y, IBRAHIM N. Prevalence and associated risk factors of calf coccidiosis in and around asela town, southeast Ethiopia[J]. Journal of Natural Sciences Research, 2016, 6(3):107-111
[4]	沈杰, 黄兵. 中国家畜家禽寄生虫名录[M]. 北京: 中国农业科学技术出版社, 2004: 8-18.
[5]	CARDIM S T, SEIXAS M, TABACOW V B D , et al. Prevalence of Eimeria spp. in calves from dairy farms in northern Paraná state, Brazil[J]. Revista Brasileira De Parasitologia Veterinária, 2018, 27(1):118-122. DOI URL
[6]	FITZGERALD P R. The economic impact of coccidiosis in domestic animals[J]. Advances in Veterinary Science and Comparative Medicine, 1980, 24:121-143.
[7]	KOUTNY H, JOACHIM A, TICHY A, et al. Bovine Eimeria species in Austria[J]. Parasitology Research, 2012, 110(5):1893-1901. DOI URL
[8]	HIEPE T. Betrachtungen zur systematischen Bekampfung von parasiten und parasitosen[J]. Monatshefte fur Veterinarmedizin, 1972.
[9]	符敖齐, 林孟初, 黄燕, 等. 扬州市郊奶牛球虫种类调查[J]. 中国兽医科技, 1987, 17(7):22-24.
	FU A Q, LIN M C, HUANG Y, et al. Investigation on coccidia species of dairy cows in Yangzhou suburb[J]. Chinese Journal of Veterinary Science and Technology, 1987, 17(7):22-24.(in Chinese)
[10]	FLORIÃO M M, LOPES B D B, BERTO B P, et al. New approaches for morphological diagnosis of bovine Eimeria species: a study on a subtropical organic dairy farm in Brazil[J]. Tropical Animal Health and Production, 2016, 48(3):577-584. DOI URL
[11]	龚振兴. 鸡球虫E. tenella叶酸合成途径关键酶PPPK-DHPS cDNA的克隆及其株间变异研究[D]. 长沙: 湖南农业大学, 2007.
	GONG Z X. The clone of chicken E. tenella PPPK-DHPS cDNA and the study of mutation in difference strains[D]. Changsha: Hunan Agricultural University, 2007. (in Chinese with English abstract)
[12]	郝力力, 李锐, 段玲, 等. 四川红原县牦牛隐孢子虫感染情况分子流行病学调查[J]. 浙江农业学报, 2016, 28(11):1842-1846.
	HAO L L, LI R, DUAN L, et al. Prevalence and molecular identification of Cryptosporidium spp. in yaks in Hongyuan County of Sichuan Province[J]. Acta Agriculturae Zhejiangensis, 2016, 28(11):1842-1846.(in Chinese with English abstract)
[13]	BANGOURA B, MUNDT H C, SCHMSCHKE R, et al. Prevalence of Eimeria bovis and Eimeria zuernii in German cattle herds and factors influencing oocyst excretion[J]. Parasitology Research, 2012, 110(2):875-881. DOI URL
[14]	曹希亮, 董辉, 黄兵, 等. 奶牛球虫流行病学调查研究[J]. 中国奶牛, 2012(3):28-31.
	CAO X L, DONG H, HUANG B, et al. Epidemiological investigation of coccidia in dairy cows[J]. Chinese Dairy Cattle, 2012(3):28-31. (in Chinese)
[15]	李豪, 郭雨佳, 刘超, 等. 我国部分地区奶牛球虫病流行病学调查[J]. 中国畜牧杂志, 2016, 52(14):35-38.
	LI H, GUO Y J, LIU C, et al. The epidemiological investigation of dairy coccidiosis in parts of our country[J]. Chinese Journal of Animal Science, 2016, 52(14):35-38. (in Chinese with English abstract)
[16]	李复煌, 于童, 苏山春, 等. 北京地区奶牛球虫感染情况调查[J]. 中国兽医杂志, 2018, 54(12):3-5.
	LI F H, YU T, SU S C, et al. Epidemiological investigation of dairy cow coccidiosis in parts of Beijing[J]. Chinese Journal of Veterinary Medicine, 2018, 54(12):3-5.(in Chinese with English abstract)
[17]	郭志宏, 彭毛, 沈秀英, 等. 青海部分地区不同年龄段牦牛球虫感染情况的调查[J]. 青海大学学报, 2017, 35(2):42-47.
	GUO Z H, PENG M, SHEN X Y, et al. Epidemiological investigation of yak coccidiosis with different ages in some regions of Qinghai[J]. Journal of Qinghai University, 2017, 35(2):42-47.(in Chinese with English abstract)
[18]	张宽宽, 齐萌, 赵爱云, 等. 库车县规模化场牛球虫感染情况及种类鉴定[J]. 中国草食动物科学, 2019, 39(3):48-50.
	ZHANG K K, QI M, ZHAO A Y, et al. Infection and species identification of bovine coccidia in large scale farms in Kuqa County[J]. China Herbivore Science, 2019, 39(3):48-50.(in Chinese)
[19]	倪晓婷, 殷铭阳, 谭启东, 等. 甘肃榆中县奶牛球虫病流行病学调查与虫种鉴定[J]. 中国畜牧兽医, 2015, 42(10):2806-2811.
	NI X T, YIN M Y, TAN Q D, et al. Epidemiologic survey and species determination of Coccidia infection in dairy cattle in Yuzhong, Gansu Province[J]. China Animal Husbandry & Veterinary Medicine, 2015, 42(10):2806-2811.(in Chinese with English abstract)
[20]	KVICEROVČ J, PAKANDL M, HYPŠA V. Phylogenetic relationships among Eimeria spp. (Apicomplexa, Eimeriidae) infecting rabbits: evolutionary significance of biological and morphological features[J]. Parasitology, 2008, 135(4):443-452. DOI URL
[21]	王鑫, 韩红玉, 姜连连, 等. 鸡球虫18S rRNA基因序列的测定与分析[J]. 生物技术通报, 2009(S1):305-309.
	WANG X, HAN H Y, JIANG L L, et al. Sequence and phylogenetic analysis of 18S rRNA gene for Eimeria in chicken[J]. Biotechnology Bulletin, 2009(S1):305-309.(in Chinese with English abstract)
[22]	崔平, 顾小龙, 方素芳, 等. 肠艾美尔球虫纯种分离及18SrDNA部分序列测定[J]. 中国动物检疫, 2010, 27(8):40-43.
	CUI P, GU X L, FANG S F, et al. The pure specie isolation and 18S rDNA part sequence of Eimeria intestinalis[J]. Chinese Journal of Animal Health Inspection, 2010, 27(8):40-43.(in Chinese with English abstract)

宁夏部分地区奶牛球虫感染情况调查与遗传进化分析

Infection investigation and genetic evolution analysis of dairy cow coccidiosis in parts of Ningxia

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