Axenic culture and identification of amastigotes from Sichuan human strain of Chinese Leishmania isolates

This work describes a simple method to yield large amounts of the isolate MHOM/CN/90/SC10H2 amastigotes-like forms in axenic cultures using promastigotes as the starting population. The isolate MHOM/CN/90/SC10H2, used in this study, belongs to an undescribed species of Leishmania endemic to hill foci in China. The method describes induced extracellular amastigote transformation of this isolate. The rounded parasite obtained in axenic culture was morphologically similar, even at the ultrastructural level, to intracellular amastigotes. Moreover, the axenic amastigotes remained viable as verified by the stage-specific genes (gp46 and p4 genes) with RT-PCR. A 70-80 kDa protein was recognized by polyclonal antibody HRP-IgG only in axenic-derived amastigotes and not in promastigotes.     

高效液相色谱法测定猪血浆及组织中喹赛多及其脱二氧代谢物

建立了测定猪血浆及肝脏、肾脏、肌肉等组织中的喹赛多及其代谢物脱二氧喹赛多的高效液相色谱(HPLC)法.血浆样用甲醇沉淀蛋白后离心取上清液进样测定.组织样先用乙腈匀浆,用正己烷脱脂后作HPLC检测.色谱柱为ODS C18柱;流动相血浆样测定为乙腈水(2080),组织样测定为甲醇水(4258),流速为1.0 mL/min;紫外检测波长305 nm.药物工作液质量浓度范围为0.005～0.500 mg/L时,血浆样品及组织样品测定条件下药物质量浓度与响应值均具良好线性关系,相关系数＞0.999.血浆中药物质量浓度为0.02、0.10、0.50 mg/L时,喹赛多及脱二氧喹赛多的回收率均大于70%,组织中药物含量为0.05、0.20、1.00μg/g时,肌肉样品的回收率均大于70%,肝脏、肾脏样品则为50%～80%.本试验条件下,喹赛多及其脱二氧代谢物的最低检出质量浓度,血浆样品分别为0.01、0.02mg/L,组织样品2种检测物均为0.025μg/g.测定了工作液3种质量浓度0.01、0.05、0.25 mg/L的仪器精密度,日内相对偏差＜8.0%,日间相对偏差＜17.0%.     

猪瘟退出强制免疫后的若干问题

猪瘟在我国被列为一类动物传染病,是危害养猪业生产的主要传染病之一。自2007年起,我国制定了以强制免疫为主的猪瘟综合防控策略,在实施了近10年的强制免疫后,我国的猪瘟群体免疫抗体合格率持续维持在较高水平,病原检出率逐年下降,猪瘟临床表现趋于和缓,流行情况趋于稳定。虽然现阶段仍存在原种场、种猪场和商品场的免疫水平参差不齐,不同省份间免疫状况差异较大,诊断、鉴别技术发展相对缓慢等情况,但强制免疫政策的实施有效控制了猪瘟的发生与流行。2017年猪瘟退出了国家强制免疫计划,改为对符合条件的养殖场实行"先打后补"的财政补助政策,这表明我国猪瘟防控取得阶段性成果,猪瘟防控进入了新的历史阶段。但防控也面临着若干新问题:一是养殖者主体责任意识不强;二是病原阳性率与临床发病率匹配度较低;三是疫情监测网络监测效率低;四是如何做好高密度免疫;五是财政支持政策需进行科学调整;六是需研究猪瘟净化方案。解决好这些问题是我国下一阶段猪瘟防控工作的关键:一要明确养殖者的防疫主体责任;二要厘清猪瘟监测阳性率与实际发病率之间关系;三要强化猪瘟监测网络建设;四要建立系统的疫苗效果评价机制;五要优化财政支持政策,充分发挥政府和市场两种资源;六要尽快研究制定符合我国国情的猪瘟净化技术方案。     

新发动物疫病检测方法研究及口岸检疫应用展望

目前我国口岸已有的一些疫病检测方法,主要是针对进境动物检疫疫病名录中提及的已知病原,而针对新发病原的检测方法较少。本文介绍了传统病原的检测方法,包括病原分离和培养、血清学方法、蛋白质免疫印迹和电子显微镜观察等,阐述了新型分子生物学检测方法,如随机引物PCR法、序列非依赖性单引物扩增技术、小RNA深度测序技术、病毒宏基因组学技术等,并对上述方法在口岸中的应用前景进行了分析。     

广东台山2012—2013年鳗鱼药残监测分析

广东台山是我国最大的鳗鱼养殖基地,其鳗鱼产品以出口为主,如活鳗的出口量约占全国活鳗总出口量的80%。药残是影响出口鳗鱼的主要因素,本研究采用液相色谱-串联质谱法(LC-MS-MS)和气相色谱-质谱法(GC-MS),对台山鳗鱼产品可能含有的100种药物残留做了两年的连续跟踪监测,结果表明,台山鳗鱼养殖业发展平稳,具有养殖科学、管理完善、用药合理、监管到位,养殖场用药少,疫病发生率低等特点。     

宁夏东部荒漠草原猪毛蒿光合特征和水分利用效率对降水变化的响应

荒漠草原植物的生长和生存受降水影响很大,本研究以宁夏东部荒漠草原猪毛蒿种群为研究对象,2017-2020年连续4年监测了降水改变条件下土壤水分特征,测定了猪毛蒿净光合速率、蒸腾速率、气孔导度和胞间CO₂浓度等光合特征,并对其内禀水分利用效率和瞬时水分利用效率进行了分析。研究表明：4年中无效降水事件(<5mm)发生次数较多,占当年降水频次的60.00%～82.54%;大雨(≥25 mm且≤49.9 mm)和暴雨等级(≥50 mm)降水事件较少,平均单次降水事件的降水量仅为3.3～6.5 mm·次^-1。不同处理各土层的土壤含水量存在较大差异,表层(10 cm)土壤含水量较次表层(20 cm)和深层(60 cm)的变幅大;次表层(20 cm)土壤平均含水量低于表层,与小降水事件发生频次较高无法入渗补给以及植物吸收消耗有关;深层(60 cm)土壤含水量变化小,较稳定。净光合速率随着光合有效辐射的增加而增大,当光合有效辐射在400～1600μmol·m^-2·s^-1时,增雨处理提高了猪毛蒿的净光合速率;在...     

Temporal aspects of the epidemic of bovine spongiform encephalopathy in Great Britain: individual animal-associated risk factors for the disease

The objectives of this study were first to determine the cumulative incidence of bovine spongiform encephalopathy (BSE) in the British cattle population from July 1986 to June 1997, secondly, to identify individual animal-associated risk factors that influenced the age of onset of clinical signs in confirmed BSE cases, and, thirdly, to assess the effectiveness of the measures introduced to control BSE during the epidemic. The analyses were based on the population of British cattle at risk, derived from agricultural census data collected between 1986 and 1996, and BSE case data collected up to June 30, 1997. The unit of interest was individual adult cattle recorded on annual agricultural censuses between June 1986 and June 1996. Univariate and multivariate survival analysis techniques were used to characterise the age of onset of clinical signs. In total 167,366 cases of BSE were diagnosed in Great Britain up to June 30, 1997. The cumulative incidence of BSE between July 1986 and June 1997 was 1.10 (95 per cent confidence interval [CI] 1.09 to 1.10) cases per 100 adult cattle at risk. Cattle from the South east, South west and Eastern regions of England had 4.26 to 5.96 (95 per cent CI 4.15 to 6.14) times as great a monthly hazard of being confirmed with BSE as cattle from Scotland. Compared with cattle born before June 1985, those born between July 1987 and June 1988 had 22.5 (95 per cent CI 22.1 to 22.8) times the monthly hazard of being confirmed with BSE, whereas those born in the 12 months after July 1988 had only 7.39 (95 per cent CI 7.24 to 7.54) times the monthly hazard of being confirmed with BSE. This reduction in hazard was directly attributable to the ban on the use of ruminant protein as a feed instituted in July 1988. Successive cohorts from 1989 to 1991 experienced further reductions in the hazard of experiencing BSE. The additional decrease in hazard observed for the 1990 cohort may be attributed to the effect of the Specified Bovine Offal ban instituted in September 1990.     

鸡传染性法氏囊病河北野毒株的分离研究

采用血清学方法、鸡胚培养、病理组织学检查方法,从河北省不同年份鸡ＩＢＤ免疫鸡群中分离和鉴定了ＨＥ９３,ＨＧ９６,ＸＮ９８和ＨＱ９９ ４个毒株,４个毒株回归鸡的死亡率分别为５０％,４０％,０％和１０％,从年份上看,ＩＢＤＶ毒力有逐渐减弱的趋势。接种鸡胚并传代,前２个毒株均在第一代时鸡胚全部死亡,且出现典型的鸡胚病变,而后２个毒株,初代培养鸡胚不死亡,当传至第４代才出现大部分死亡和鸡胚病变。ＨＥ９３、     

香蕉茎叶粉固态发酵条件优化及鹅对其养分利用率的研究

本试验旨在研究应用固态发酵技术改善香蕉茎叶粉养分组成的工艺参数及鹅对其养分的利用率。选择米曲霉和产朊假丝酵母作为发酵菌种,逐步探究5个因素(单菌种发酵、硫酸铵添加量、米曲霉和产朊假丝酵母接种比例、混合菌液接种量)对发酵后香蕉茎叶粉蛋白质含量的影响。利用正交设计筛选最优发酵温度、底物水分和发酵时间的组合。结果显示:米曲霉和产朊假丝酵母单独发酵均能显著或极显著提高发酵后香蕉茎叶粉的粗蛋白质含量(P0.05或P0.01)。添加硫酸铵能显著或极显著提高发酵后香蕉茎叶粉的真蛋白质含量(P0.05或P0.01),其中添加2%硫酸铵组的真蛋白质含量最高。米曲霉∶产朊假丝酵母接种比例为2∶1的处理发酵后香蕉茎叶粉的蛋白质净增加量显著或极显著高于比例为1∶1、1∶3和3∶2的处理(P0.05或P0.01)。正交试验结果显示,以4%的接种量、接种比例为2∶1(米曲霉∶产朊假丝酵母)、2%的硫酸铵添加量,在基质水分为50%,30℃的环境下发酵4 d效果最佳。经过该工艺发酵后的香蕉茎叶粉粗蛋白质含量提高了33.82%,氨基酸分析结果显示,除赖氨酸和精氨酸外其余的15种氨基酸含量均有不同程度的提高。马冈鹅的代谢试验结果显示,发酵后香蕉茎叶粉中的粗蛋白质利用率提高了52.66%,极显著高于发酵前(P0.01);此外,代谢能和能量利用率也都略有提高(P0.05)。由此可见,经过该发酵工艺发酵后的香蕉茎叶粉营养价值不仅得到了改善,也促进了鹅对其养分的消化吸收。     

The Epidemiology and Characteristics of Q fever and Co‐infections with Scrub Typhus,Murine Typhus or Leptospirosis in Taiwan: A Nationwide Database Study

Q fever (QF) is a worldwide zoonosis associated with outbreaks. Only a few nationwide studies regarding the surveillance and epidemiology of human QF have been reported. Although QF is endemic in Taiwan, a nationwide database investigation of the epidemiology and characteristics of QF and its associations with scrub typhus (ST), murine typhus (MT) and leptospirosis (LS) has never been reported. We analysed nationwide databases of suspected QF, ST, MT and LS cases from October 2007 to December 2014 obtained from the Centers for Disease Control, Taiwan. A total of 468 (4.2%) QF cases were identified among 11 109 suspected QF cases. QF cases were mainly distributed in the southern and Kaohsiung–Pingtung regions but rarely in the eastern region. Compared to non‐QF cases, QF cases had significantly higher percentages of males (88.7 versus 66.2%) and high‐risk occupations (farming, animal husbandry or veterinary medicine) (16.2 versus 10.5%). But the percentages of specific animal contact, including cattle (0.6 versus 0.8%) and goats (0.9 versus 1.0%), were low in both. The majority of suspected QF cases (89.4%) were simultaneously suspected with ST, MT or LS, and the combinations of suspected diseases differed between regions. The number of suspected QF cases from the eastern region decreased since 2009, which was not observed in other regions. A total of 1420 (12.8%) cases had confirmed diseases, including QF (453, 4.1%), QF+ST (7, 0.06%), QF+MT (4, 0.04%), QF+LS (4, 0.04%), MT (186, 1.7%), ST (545, 4.9%), ST+LS (11, 0.1%) and LS (210, 1.9%). Compared to cases of unknown disease, QF cases had larger percentages of high‐risk occupations (16.2 versus 9.6%) but similar histories of animal contact (29.8 versus 25.1%). QF is an endemic disease in southern Taiwan. It is difficult to differentiate QF from ST, MT or LS only by high‐risk occupations and history of animal contact, and co‐infection of QF with these diseases should be considered.