为了改善地克珠利(Diclazuril)的水溶性问题,用氧化石墨烯(GO)作为药物载体制备地克珠利-氧化石墨烯纳米缓释剂,通过高效液相色谱仪(HPLC)测定石墨烯的载药量,以及通过存活率、相对增重率、盲肠病变值、卵囊值和抗球虫指数(ACI)等指标对制备药物进行抗柔嫩艾美耳球虫的疗效评价.结果显示:氧化石墨烯对地克珠利的最大负载量可达2.2 mg/mg,稀释为用药浓度范围时可在水中均匀分布;抗柔嫩艾美耳球虫的疗效评价显示:地克珠利-氧化石墨烯高、中剂量组的相对增重率(94.96％、89.9％)和抗球虫指数(ACI=192.96、179.40),均高于地克珠利药物对照组的相对增重率(83.36％)和抗球虫指数(ACI=163.36).本研究结果表明,制备的地克珠利-氧化石墨烯纳米缓释剂能有效改善地克珠利在水中的溶解问题,且可作为柔嫩艾美耳球虫的治疗药物,以中剂量组浓度1 mg/mL为最适推荐用量.

Cryptosporidium spp. are important foodborne and waterborne pathogens in humans and animals, causing diarrheal diseases. Cattle are one of the reservoirs of Cryptosporidium infection in humans. However, data on the occurrence of Cryptosporidium spp. in cattle in Yunnan Province remains limited. A total of 700 fecal samples were collected from Holstein cows (n = 442) and dairy buffaloes (n = 258) in six counties of Yunnan Province. The occurrence and genotypes of Cryptosporidium spp. were analyzed using nested PCR and DNA sequencing. Furthermore, the C. andersoni isolates were further analyzed using multilocus sequence typing (MLST) at four gene loci (MS1, MS2, MS3, and MS16), and the C. parvum isolate was subtyped by 60-kDa glycoprotein (gp60) loci. The occurrence of Cryptosporidium spp. in Holstein cows and dairy buffaloes was 14.7% (65/442) and 1.1% (3/258), respectively. Of these positive samples, 56 Holstein cow samples represented C. andersoni, four Holstein cow samples represented C. bovis, three Holstein cow samples represented C. ryanae, and one represented C. parvum. Meanwhile, only three dairy buffalo samples represented C. ryanae. MLST analysis of subtypes of C. andersoni detected four subtypes, including A5A4A2A1 (n = 7), A4A4A4A1 (n = 7), A1A4A4A1 (n = 2), and A4A4A2A1 (n = 1). One C. parvum isolate was identified as the IIdA18G1 subtype. These results revealed the high occurrence and high genetic diversity of Cryptosporidium spp. in Holstein cows in Yunnan Province, enriching the knowledge of the population genetic structure of Cryptosporidium spp. in Yunnan Province.

目的 了解云南省部分地区奶牛芽囊原虫感染情况及其基因亚型,为该病的防控提供数据支持.方法 于2019年6月-2020年8月从昆明、大理、保山地区采集荷斯坦奶牛和奶水牛粪样,提取粪样DNA,巢式PCR扩增芽囊原虫小亚基(SSU) rRNA基因,测序后在NCBI上进行BLAST比对,用MEGA 6.0软件以邻接法构建系统进化树.采用SPSS 20.0与SAS 9.1软件对不同地区、季节、养殖方式、品种、性别和发育阶段奶牛感染率进行统计分析,对差异有统计学意义的因素进行二元logistic回归分析.结果 共采集奶牛粪样841份,其中65份SSU rRNA基因扩增阳性,扩增片段长1100 bp.奶牛芽囊原虫总感染率为7.7％ (65/841,95％CI:5.92％～9.53％),其中,昆明、保山、大理地区的感染率分别为12.9％(32/248,95％CI:8.73％～17.08％)、8.1％ (19/236,95％CI:4.58％～11.52％)和3.9％ (14/357,95％CI:1.91％～5.94％),不同地区之间感染率差异有统计学意义(x2=16.601,P＜0.01);夏、秋、冬季的感染率分别为9.4％(56/599,95％ CI:7.86％～37.30％)、8.3％ (8/96,95％CI:2.80％～13.86％)和0.7％(1/146,95％CI:0～2.02％),不同季节之间感染率差异有统计学意义(x2=12.411,P＜0.05);圈养、放养的感染率分别为8.3％ (58/702,95％CI:6.23％～10.30％)、5.0％(7/139,95％CI:1.40％～8.67％),二者差异无统计学意义(x2=1.693,P＞0.05);荷斯坦奶牛、奶水牛的感染率分别为8.4％ (41/490,95％CI:5.92％～10.82％)、6.8％ (24/351,95％CI:4.20％～9.48％),二者差异无统计学意义(x2=0.671,P＞0.05);公牛、母牛的感染率分别为11.1％(10/90,95％CI:4.62％～17.60％)、7.3％ (55/751,95％CI:5.46％～9.19％),二者差异无统计学意义(x2=1.617,P＞ 0.05);断奶前犊牛、断奶后犊牛、育成牛、成年牛的感染率分别为2/18 (95％CI:0～25.63％)、26.2％ (11/42,95％CI:12.89％～39.49％)、15.1％ (11/73,95％CI:6.86％～23.28％)、5.8％ (41/708,95％CI:4.07％～7.51％),不同阶段间感染率差异有统计学意义(x2=29.604,P＜ 0.01).二元logistic回归分析结果显示,季节(P＜0.05)和发育阶段(P＜0.01)是奶牛感染芽囊原虫的独立影响因素.序列比对分析结果显示,1份粪样的SSU rRNA基因序列与GenBank登录号KX618192(ST3亚型)的序列一致性为96.3％,35份与MH104059、KC148207、MH358363(ST10亚型)的序列一致性为97.3％～～99.7％,23份与MT453998、MT453999(ST12亚型)的序列一致性为98.7％～～ 100％,6份与MH358361、MH358362、KC148205、KC148206(ST14亚型)的序列一致性为98.7％～100％.系统进化树分析结果显示,本研究中的芽囊原虫分别汇聚在ST3、ST10、ST12和ST14亚型分支上,各亚型的构成比分别为1.5％(1/65)、53.9％ (35/65)、35.4％ (23/65)和9.2％ (6/65).结论 云南省奶牛存在芽囊原虫感染,ST3和ST12基因亚型具有人兽共患风险,应该加强对该病原体的防控.

为了解中甸牦牛消化道寄生虫感染情况,为牦牛寄生虫病提供有效的防治措施,从昆明小哨和香格里拉市随机采集106份新鲜粪便样品,采用饱和盐水漂浮法和自然沉淀法共检测出49份样品有寄生虫感染,平均感染率为46.23％.其中,线虫感染率为40.57％,吸虫感染率为16.98％,线虫和吸虫混合感染率为11.32％.调查表明,该地区中甸牦牛肠道寄生虫感染情况较为严重,以线虫感染为主.中甸牦牛对驱虫药比较敏感,驱虫效果明显,感染率显著低于自由放牧牛群.因此,生产中应加强采取防治措施,提高对寄生虫病的认识和防范,以确保牦牛养殖的健康可持续发展,提质增效.

Enterocytozoon bieneusi is a fungus-like protist parasite that can cause diarrhea and enteric diseases. The infection of E. bieneusi has been reported in many host species, including cattle and humans. However, information on prevalence and genotype distribution of E. bieneusi in dairy cattle in Yunnan province in China is still absent. In this study, 490 Holstein Cows and 351 dairy buffalo fecal samples were collected from three regions in Yunnan province, China. By using nest-PCR that targets the internal transcribed spacer (ITS), we found that the prevalence of E. bieneusi was 0.59% (5/841). DNA sequence analysis showed that five E. bieneusi genotypes were identified in this study, including two novel genotypes, YNDCEB-90 and YNDCEB-174, and three known genotypes (I, J, BEB4). Phylogenetic analysis revealed that two novel genotypes, YNDCEB-90 and YNDCEB-174, were clustered into Group 1, representing the zoonotic potential. The remaining genotypes I, J, and BEB4, which are the most frequent genotypes of E. bieneusi infection in cattle and lead to E. bieneusi infection in humans, belonged to Group 2. Although the lower prevalence of E. bieneusi was detected in dairy cattle in Yunnan province, it indicates that dairy cattle should be considered to be one of the potential hosts for transmitting E. bieneusi to humans. These findings are important for the development of effective prevention strategies for microsporidiosis.

为明确云南地区部分养鸡场住白细胞虫的流行情况,试验采用巢式PCR方法对970份鸡血样扩增住白细胞虫的线粒体细胞色素b基因(cytb).结果显示:34份样本为住白细胞虫阳性,阳性检出率为3.51％(34/970);同源性分析表明,阳性样本均为卡氏住白细胞虫(Leucocytozoon caulleryi).对阳性样本进行不同性别、日龄、品种和地区等因素比对分析,发现地理区域、日龄、品种间的阳性率均存在极显著相关性(P＜0.001),而性别间无显著相关性(P＞0.05).本试验结果可为云南养鸡场住白细胞虫病的防治提供参考.

Blastocystis is an enteral eukaryote that omnipresent existence in animals and humans global. The objective of the current study investigated the frequency and subtypes to evaluate risk factors for Blastocystis infection in pigs in southwestern China. The 866 fecal samples were collected from farmed pigs in Yunnan province, which were determined by the molecular method of the nested PCR and phylogenetic analysis established on the small subunit rDNA (SSU rDNA) gene fragment. A total of 433 specimens were detected Blastocystis positive, and the prevalence of is 50.0% (433/866). The significant differences in prevalence were also observed in pigs from regions (P < 0.01, df = 8, χ2 = 29.17) and age groups (P < 0.01, df = 2, χ2 = 66.6). Older pigs had more Blastocystis infection than younger ones. Moreover, three zoonotic subtypes were identified by DNA sequence analysis, included ST1 (7.39%, 32/433), ST3 (4.39%, 19/433) and ST5 (88.22%, 382/433). The results suggested a high prevalence and diversified subtypes in farmed pigs in Yunnan province, southwestern China, which indicated pigs are most likely to potential reservoirs and zoonotic transmission of Blastocystis for human infection. To our knowledge, the study is the first large-scale systematic statement of Blastocystis colonization in Yunnan province, Southwestern China, which contributed to the in-depth study of genetic characteristics and prevention, control and treatment of Blastocystis in pigs in Yunnan province and other regions.

Amebiasis is a worldwide parasitic zoonosis, with symptoms of abdominal discomfort, indigestion, diarrhea, and even death. However, limited information about the prevalence of Entamoeba spp. in experimental nonhuman primates (NHPs) in southwestern China is available. The objective of the current study was to investigate the frequency and species identity of Entamoeba to evaluate potential zoonotic risk factors for Entamoeba spp. infection in experimental NHPs. A total of 505 fecal samples were collected from NHPs (macaques) and analyzed by PCR analysis the small subunit rRNA (SSU rRNA) gene of Entamoeba spp. Forty-seven specimens were positive for Entamoeba spp., and the prevalence of Entamoeba spp. was 9.31% (47/505). Significant differences in the prevalence rates among the three breeds (P = 0.002 < 0.01, df = 2, χ2 = 12.33) and feed types (P = 0.001 < 0.01, df = 1, χ2 = 10.12) were observed. Altogether, four Entamoeba species, including E. dispar (57.44%), E. chattoni (29.78%), E. histolytica (6.38%), and E. coli (6.38%), were identified by DNA sequence analysis. The results suggested a low prevalence but high diversity of Entamoeba species in experimental NHPs in Yunnan Province, southwestern China. Results of this study contribute to the knowledge of the genetic characteristics of Entamoeba spp. in NHPs.

[目的]为调查云岭牛肠道寄生虫的感染情况,以其提供有效的防治措施.[方法]从小哨云岭牛核心育种场和楚雄锦达牧业扩繁场采集382份新鲜粪便样品,采用饱和盐水漂浮法和自然沉淀法检测寄生虫感染情况.[结果]共检测出89份样品存在寄生虫感染,平均感染率为23.30％.分别检出了线虫、球虫和吸虫的感染,其感染率分别为17.28％、7.07％、0.26％,混合感染率为1.31％.云岭牛核心育种场寄生虫感染率为31.25％,楚雄锦达牧业扩繁场为15.26％;自由放牧云岭牛寄生虫感染率为36.05％,圈养云岭牛为19.59％;不同养殖场、不同养殖方式的云岭牛之间寄生虫感染存在极显著差异(P＜0.01),而不同性别和年龄的云岭牛之间无显著性差异(P＞0.05).[结论]云岭牛寄生线虫感染较为突出,应加强综合防治以促进肉牛养殖业的健康可持续发展.

为了解武定县牛肠道寄生虫的感染情况,以及为其提供有效的防治措施,从武定县某养殖户随机采集14份黄牛和10份水牛粪便样品,通过饱和盐水漂浮法和自然沉淀法检查寄生虫感染情况,其中,线虫卵检出14份,检出率为58.33％,球虫卵囊检出17份,检出率为70.83％,吸虫卵检出14份,检出率为58.33％,混合感染检出19份,检出率为79.17％.调查结果表明,武定县牛肠道寄生虫感染情况十分严重,以线虫、球虫和吸虫感染为主,需加强综合防治措施,以确保养殖户的经济效益.

芽囊原虫(Blastocystis spp.)病是一种人兽共患寄生虫病,广泛分布于世界各地.曾经长期被认为是一种对人体无害的酵母,近年来大量证据表明,芽囊原虫是一类可寄生在人和灵长类等多种动物的肠道内可致病的原虫.随着研究的深入,非人灵长类动物芽囊原虫病逐渐引起人们的关注,非人灵长类动物和人类具有相似的基因背景和表型特征而成为研究人类疾病中的理想动物模型.本文从流行病学、 基因分型等方面总结了国内外芽囊原虫在非人灵长类动物中感染情况和不同国家之间基因型差异的研究进展,探讨了非人灵长类动物芽囊原虫病的公共健康危害和防治方法,为养殖人员防治芽囊原虫提供参考.芽囊原虫的生活史尚不完全清楚,其发病机制也尚未明确;对其生活史和发病机制研究的深入,将有助于防治芽囊原虫病.

为了鉴定送检池鹭所感染的消化道线虫种属关系,通过对虫体、 虫卵形态学观察和虫体cox1基因的序列分析进行虫体鉴定,即采用PCR扩增线粒体DNA中细胞色素I(cox1)基因,获得有效序列进行NCBI-Blast在线比对,并构建遗传系统进化树.结果显示,池鹭消化道线虫cox1序列长度为443 bP,与狮弓首蛔虫同源性为99％,系统进化分析显示与狮弓首蛔虫位于同一分支.结合形态学观察结果表明,池鹭所感染的线虫属于狮弓首蛔虫,鉴定结果可为池鹭消化道线虫的防治提供参考.

在云南省武定县某笼养和放养混合养殖的鸡场采样38只成年活鸡,用完全剖检法对其进行体外寄生虫和消化道寄生虫的检测,以期了解武定鸡寄生虫的感染情况,以便制定合适的驱虫方案.检测结果表明,32只鸡检出羽虱,检出率84.2％;13只检出异刺线虫,检出率34.2％;8只检出蛔虫,检出率21.1％;2只检出绦虫,检出率5.3％.结果显示,该鸡场有寄生虫感染,且十分严重,出现这种情况可能与鸡场的饲养方式和驱虫方案有关.