Background: Cryptosporidium is a globally distributed zoonotic protozoan parasite in humans and animals. Infection is widespread in dairy cattle, especially in calves, resulting in neonatal enteritis, production losses and high mortality. However, the occurrence of Cryptosporidium spp. in pre- and post-weaned calves in Yunnan Province remains unclear. Methods: We collected 498 fecal samples from Holstein calves on 10 different farms in four regions of Yunnan Province. Nested PCR and DNA sequencing were used to determine the infection, species and genotypes of Cryptosporidium spp. in these animals. Results: The overall occurrence of Cryptosporidium spp. in Holstein calves was 32.9% (164/498), and the prevalence in pre- and post-weaned calves was 33.5% (106/316) and 31.9% (58/182), respectively. Four Cryptosporidium species were identified in these animals, namely C. bovis (n = 119), C. parvum (n = 23), C. ryanae (n = 20) and C. andersoni (n = 2). Based on sequencing analysis of the 60 kDa glycoprotein gene of C. bovis, C. parvum and C. ryanae, six subtypes of C. bovis (XXVIe, XXVIb, XXVIf, XXVIa XXVIc and XXVId), two subtypes of C. parvum (IIdA19G1 and IIdA18G1) and four subtypes of C. ryanae (XXIf, XXId, XXIe and XXIg) were identified. Conclusions: These results provide essential information to understand the infection rate, species diversity and genetic structure of Cryptosporidium spp. populations in Holstein pre-weaned and post-weaned calves in Yunnan Province. Further, the presence of IIdA18G1 and IIdA19G1 in C. parvum implies significant animal and public health concerns, which requires greater attention and more preventive measures.

Cyclospora spp. is a food-borne intestinal protozoan, which is widely distributed in the world and poses the risk of zoonosis. In order to reveal the prevalence of Cyclospora spp. in Holstein cattle in partial areas of the Yunnan Province, 524 fresh fecal samples of Holstein cattle were collected from Dali, Kunming, Chuxiong, and Qujing in Yunnan Province. A nested PCR amplification of the small subunit (SSU) rRNA gene of Cyclospora spp. was carried out, and the products of the nested PCR were further analyzed by restriction fragment length polymorphism (RFLP) using Bsp E Ⅰ. The results of the present study showed that 13 samples were positive for Cyclospora spp., and the total infection rate of Cyclospora sp. was 2.48%. The infection of Cyclospora spp. was detected in Dali, Qujing, and Chuxiong. Chuxiong showed the highest infection rate (5.71%), and infection rate in Dali and Qujing was 2.19% and 3.16%, respectively. Interestingly, the infection of Cyclospora spp. was not detected in Kunming. The infection of Cyclospora spp. showed no significant differences among different regions (p > 0.05). Cyclospora sp. infection was detected in all ages and sexes, but the differences were not significant (p > 0.05). Sequencing and phylogenetic analysis showed that five Cyclospora spp. samples were closely related to the Cyclospora spp. of humans, and the others were closely related to the Cyclospora spp. of bovines. The results of the present study suggested that there was an infection of Cyclospora spp. in Holstein cattle in the Yunnan Province, and the Cyclospora spp. showed a risk of zoonosis. Thus, the prevention and control of Cyclospora spp. should be strengthened in the Yunnan Province, China. The results of this investigation provide data references for the further research of Cyclosporiasis in Holstein cattle in the Yunnan Province.

[Objective] In order to understand the infection of intestinal parasites of beef cattle in Mang City and provide reference for the prevention and treatment of parasitic diseases. [Method] 191 fresh feces samples were collected from Dehong beef farm in Yunnan province, and the infection of parasites was examined by the methods of saturated brine floating and natural precipitation. [Result] A total of 124 samples were found to be infected with parasites, with a total infection rate of 64.92%. Coccidia, nematode and trematode were detected, with an infection rate of 30.89%, 4.19% and 15.71% respectively, and a mixed infection rate of 14.41%. There was significant difference in parasite infection among beef cattle of different breeds（P＜0.05）, and there was no significant difference in parasite infection among beef cattle of different farms, genders, and ages（P＞0.05）. [Conclusion] Cattle in Mang City are seriously infected with parasites, especially coccidia. Regular deinsectization and comprehensive control should be strengthened to promote the healthy breeding of beef cattle.

Blastocystis is an opportunistic pathogenic protozoan with worldwide distribution,to identify the infection status and the genotypes of Blastocystis that infected Struthio camelus in the Struthio camelus breeding profes-sional cooperatives in Yunnan province,90 faecal samples were collected to amplify the SSU rRNA gene of Blasto-cystis by nested PCR.The result showed that 62 samples were identified as the positive of Blastocystis;The detec-tion rate of Blastocystis was 68.89％(62/90),the highest infection rate was 86.36％(19/22)at 90 days of age group and the lowest infection rate was 6.25％(20/32)at 60 days of age group,there was no significant difference in the infection rate of Blastocystis among different age groups(P＞0.05,X2=6.67,df=3);Based on the sequence analysis of SSU rRNA gene,three subtypes of Blastocystis(ST5,ST7 and ST20)were identified.ST5 is the domi-nant subtype,and ST5,ST7 are the subtypes of zoonosis.This results enriched the genotyping data of Blastocystis from Struthio camelus in Yunnan province as well as provided reference for the prevention and control of Blasto-cystis.

[目的]对云南某动物园袋鼠粪样中分离到的1株隐孢子虫分离株进行种/基因型的分子鉴定,为云南动物园的袋鼠隐孢子虫病防控提供理论依据.[方法]针对隐孢子虫小亚基核糖体RNA基因序列设计引物,进行巢式PCR扩增,并对扩增产物进行测序和序列比对;使用DNAMAN和MEGA7.0等软件进行同源性分析并构建系统发育进化树,以确定该袋鼠源分离株与其他源隐孢子虫间的亲缘关系.[结果]成功扩增出隐孢子虫阳性片段.BLAST结果表明:获得的袋鼠源隐孢子虫分离株与鼠隐孢子虫(Cryptosporidium muris)一致,同源性可达100％.种系发育进化树表明:该分离株与鼠隐孢子虫处于同一分支,亲缘关系最近,属于胃寄生隐孢子虫的鼠隐孢子虫.[结论]该园袋鼠存在隐孢子虫感染,鉴定为具有人兽共患潜能的鼠隐孢子虫.本研究首次报道了中国袋鼠感染鼠隐孢子虫.

为了解元江县部分山羊肠道寄生虫感染状况,采用饱和食盐水漂浮法结合麦克马斯特氏法,对该县2个采样点6个不同养殖场中采集的168份山羊粪样进行了寄生虫检测及阳性粪样虫卵计数,并采用巢氏PCR方法对粪样中毕氏肠微孢子虫、芽囊原虫和隐孢子虫进行检测.结果显示,该地区山羊肠道寄生虫总感染率为44.64％,以线虫和球虫感染为主,球虫感染率为44.64％(75/168),每克粪便样品中的球虫卵囊数(OPG值)在0～23800,平均为888.33;线虫感染率为3.57％(6/168),每克粪便样品中的线虫虫卵数(EPG值)在0～200,平均为7.14;混合感染率为3.57％(6/168).不同采样地间感染率差异极显著(P<0.01),不同年龄段、性别间感染率差异不显著(P>0.05).巢式PCR检测结果显示,毕氏肠微孢子虫、芽囊原虫感染率分别为4.76％(8/168)、0.60％(1/168),未检测到隐孢子虫感染.调查结果表明元江县山羊肠道寄生虫感染已普遍存在,可为元江县山羊肠道寄生虫病的科学防控提供参考.

Giardia duodenalis is an important zoonotic protozoon, which can infect a variety of animals, causing diarrhea and even death of animals or humans. Dairy cattle have been implicated as important sources of human G. duodenalis. However, the information about the prevalence and genetic diversity of G. duodenalis in dairy cattle in China's Yunnan Province remains limited. This study investigated the occurrence and multilocus genotyping of G. duodenalis of Holstein cattle in Yunnan Province, China. A total of 524 fresh fecal samples of Holstein cattle were randomly collected from 8 farms in Yunnan. In this study, 27.5% (144/524) of tested samples were positive for G. duodenalis infection. The highest infection ratio was found in preweaned calves (33.7%), and the infection rates of postweaned calves, growing cattle, and adult cattle were 24.5%, 23.0%, and 17.3%, respectively. The sequence analysis of SSU rRNA gene showed that the predominant assemblage of G. duodenalis in this study was assemblage E (97.9%, 141/144), whereas assemblage A was identified only in three samples (2.1%, 3/144). All G. duodenalis-positive samples were further assayed with nested polymerase chain reaction (PCR) targeting β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh) genes, and 87, 41, and 81 sequences were obtained, respectively. Mixed infection of assemblages A and E of G. duodenalis was detected in three samples. Multilocus genotyping yielded 23 multilocus genotypes (MLGs). This is the first study that reveals the prevalence data of G. duodenalis in Holstein cattle in Yunnan Province, and the results of this study provided baseline data for the prevention and control of G. duodenalis infection in Holstein cattle in Yunnan Province, China.

为了解云南大理地区奶牛内阿米巴原虫的流行现状,采集大理3个地区的散养和规模化养殖奶牛新鲜粪便样品共计113份,使用PCR方法对内阿米巴原虫的18S rRNA基因位点进行扩增分析.结果显示,奶牛内阿米巴原虫总感染率为30.09％;宾川、洱源和巍山3个地区平均感染率分别为32.20％、27.78％、27.78％;散养和规模化养殖的奶牛内阿米巴的感染率分别为25.00％、30.69％;测序及种系发育关系分析结果显示,34份阳性样品中共有32份样品为Entamoeba bovis阳性,2份为Entamoeba sp.MG107/BEL阳性.此次调查的3个地区均存在不同程度的内阿米巴原虫感染,不同方式养殖的奶牛,内阿米巴原虫感染率无显著差异,E.bovis为云南省奶牛感染内阿米巴的优势虫种.

为了解云南贡山县独龙鸡肠道寄生虫的感染情况,本试验在怒江贡山县丙中洛地区收集了67份独龙鸡粪,采用饱和食盐水漂浮法和显微镜观察,共检出42份存在寄生虫感染,平均感染率为62.69％,检出了线虫卵和球虫卵囊,线虫感染率为62.69％,球虫感染率为5.97％,其中4份样品存在线虫和球虫的混合感染,混合感染检出率为5.97％.由此可见,线虫感染率最高,为本鸡场主要感染虫种,应重点加强线虫的防治工作.

为了解乌鲁木齐市周边绵羊养殖农户的沙门菌污染及其耐药性和致病力,采集乌鲁木齐市周边5家农户绵羊源的285份样品,按照国家标准GB 4789.4-2016进行沙门菌的分离培养,结合PCR方法鉴定,检测了19种毒力基因的编码情况,采用琼脂稀释法测定了13种抗菌药物的耐药情况.共分离鉴定出62株沙门菌,分离率为24.03％,其中粪便、肛拭子、饲料、体表拭子的检出率依次为13.82％ (10/78),30.21％(29/96),13.33％(2/15),30.43％(21/69);19种毒力基因检出了16种,有沙门菌毒力岛上的sipB (100.00％)、invJ (98.39％)、mogA (87.10％)、spiA(98.39％)、sifA(87.10％)、ssaQ(100.00％)、sseL(66.13％)、mgtC(80.65％)、araB (98.39％)、sopB (93.55％),毒力质粒上的spvA(16.13％)、spvR(16.13％),菌毛上的lpfC(87.10％)、bcf(66.13％)以及肠毒素stn(96.77％);菌株的耐药率依次为:阿莫西林-克拉维酸钾和四环素(93.5％)、氨苄西林(91.9％)、头孢噻呋(80.6％)、多黏菌素E(79.0％)、头孢曲松钠(69.4％)、头孢西丁钠(66.1％)、卡那霉素(64.5％)、庆大霉素(59.7％)、恩诺沙星(40.3％)、氯霉素和阿奇霉素(1.6％);沙门菌的多重耐药最高达到10耐,依次从高到低是8耐(23.3％)、9耐(21.7％)、10耐(16.7％)、5耐和6耐(11.7％)、7耐(8.3％)、3耐(5.0％)、4耐(3.3％).乌鲁木齐市周边散农户羊场的沙门菌污染率较高,具有较高的致病性,耐药性现象严重.

鸡沙门氏菌为革兰氏阴性、兼性厌氧、无芽孢的小杆菌,具有血清型繁多、分布广、传染性强以及危害大等特点,一直是世界各地人类食物性中毒以及畜禽感染发病的重要致病菌.2018年7月,兴农动物医院接诊了1例5日龄雏鸡大量死亡病例,通过病理剖检和实验室检测诊断为鸡沙门氏菌病,结合药物敏感性实验使用阿莫西林和乳酸环丙沙星饮水,3 d后成功治愈该病.通过病例的诊断和治疗,可为临床上治疗鸡沙门氏菌病提供借鉴.