基于18S rRNA和HSP70基因序列的隐孢子虫种系发育分析

为阐明河南区域隐孢子虫分子流行病学特点,用PCR技术扩增分离虫株的18S rRNA基因全序列和HSP70基因序列,并对扩增片段进行测序。用PAUP 4.0和TREEPUZZLE 4.1构建进化树,试图从分子水平证明河南省不同地区不同宿主来源隐孢子虫的遗传特征,以阐明隐孢子虫病的分子流行病学特点。通过18S rRNA基因全序列和HSP70基因序列分析,其结果：河南人源隐孢子虫分离株为Cryptosporidium parvum鼠基因型;河南鹿源隐孢子虫分离株为C. parvum鹿基因型;河南猪源隐孢子虫的2个分离株均为C. parvum猪基因I型,即C. suis;河南鹌鹑源的隐孢子虫2个分离株分别为C. baileyi和C. meleagridis;河南乌鸡源隐孢子虫和鸵鸟源隐孢子虫分离株均为C. baileyi;河南牛源隐孢子虫分离株为C.andersoni。     

基于ITS-1基因序列的隐孢子虫种群分类

本试验以内转录间隔区1(ITS-1)基因作为研究对象,用PCR技术分别对广东(GD)禽源、广东(GD)和安徽(AH)牛源3个隐孢子虫分离株ITS-1基因进行扩增、克隆、序列测定和分析.将扩增序列用DNAStar(4.0)和ClustalX1.81软件与GenBank上登录的参考序列比对,用Phylip3.67构建种群发育进化树,以确定分离株与其他隐孢子虫虫种之间的亲缘关系.通过对ITS-1基因全序列分析,结果表明:禽源隐孢子虫GD分离株为Cryptosporidium baileyi,牛源隐孢子虫GD分离株和AH分离株均为C.andersoni.因此,ITS-1基因可作为隐孢子虫分离株种群分类的遗传标记,从而为隐孢子虫属的种间鉴定以及进一步的分子流行病学调查和分子诊断学研究奠定了基础.     

不同禽源贝氏隐孢子虫分离株对鹌鹑致病性的比较研究

本研究旨在观察不同禽源贝氏隐孢子虫(Cryptosporidium baileyi)对鹌鹑的致病性差异。采用光镜、扫描电镜等技术研究了鸡源、鸭源、鹌鹑源、白文鸟源、鸵鸟源贝氏隐孢子虫分离株对10日龄鹌鹑的致病性。结果表明,感染鹌鹑均出现腹泻和呼吸道症状,排卵囊持续期11~18 d,虫体寄生于喉头、气管、法氏囊和泄殖腔。其中鸭源C.baileyi感染组鹌鹑发病率和死亡率分别为82.5%、25%,鸡源C.baileyi(郑州株)感染组发病率和死亡率分别为77.5%、12.5%,鹌鹑源C.baileyi感染组发病率、死亡率分别为37.5%和17.5%。本研究结果提示,不同禽源C.baileyi对鹌鹑有一定致病性,其致病性强弱存在差异。     

应用巢式PCR检测隐孢子虫

应用巢式聚合酶链反应(Nested PCR)建立了一种检测隐孢子虫(Cryptosporidium)的方法。试验中隐孢子虫卵囊纯化采用庶糖密度梯度离心法,以液氮-热水浴反复冻融及酚-氯仿抽提冷乙醇沉淀法制备模板DNA,根据隐孢子虫18S rRNA序列高度保守区设计2对引物,建立Nested PCR诊断方法。该方法特异性强,可检出牛源微小隐孢子虫(C.parvum)、羊源微小隐孢子虫(C.parvum)、牛源安氏隐孢子虫(C.andersoni)、鸡源贝氏隐孢子虫(C.baileyi)及猪源隐孢子虫(C.suis);敏感性高,该方法最低核酸DNA检测量达到10fg。初步应用结果表明,所建立的Nested PCR方法适合于隐孢子虫病的诊断和分子流行病学调查。     

隐孢子虫（Cryptosporidium）病毒的鉴定及其特性

根据已发表的隐孢子虫病毒的序列,设计合成2对引物,对小球隐孢子虫(Cryptosporidiumparvum)、鼠隐孢子虫(C.muris)、贝氏隐孢子虫(C.baileyi)、火鸡隐孢子虫(C.meleagridis)进行RT-PCR扩增,结果发现仅小球隐孢子虫(C.parvum)含有大小约为1.7×103nt和1.3×103nt2个片段的dsRNA病毒.将其PCR产物连接到pMD18-T载体上进行克隆测序,经与GenBank比较,含这2个片段的dsRNA病毒与已发表的该虫病毒的同源性分别为96%和98%.电泳鉴定发现,该病毒对低浓度RNA酶不敏感,且不能被DNA酶降解.依赖RNA的RNA聚合酶活性测定结果表明,该病毒具有该聚合酶的活性.电镜观察未发现存在病毒样粒子.     

火鸡隐孢子虫18S核糖体DNA部分序列测定与系统发育分析

从长春地区鸭的粪便中分离纯化了火鸡隐孢子虫（Cryptosporidium meleagridis）卵囊，根据隐孢子虫18S rDNA基因序列设计合成引物，用PCR扩增了卵囊基因组DNA大小586bp的片段，PCR产物经电泳鉴定后用试剂盒回收纯化，纯化后PCR产物直接测序，将测得的序列用Dnastar软件分析并与国外已发表的相应序列进行了同源性比较，并绘制了系统发育进化树。结果初步建立了火鸡隐孢子虫的PCR检测方法，序列分析显示长春外国语源火鸡隐孢子虫与国外9株隐孢子虫相应序旬同源性在82.7%-99.8%之间，其中与国外火鸡源火允隐孢子虫相应序列同源性为85.3%；与C.felis同源性最低，与C.muris同源性最高。本研究为火鸡隐孢子虫病诊断及该病分子流行病学研究打下了良好基础。     

不同禽源贝氏隐孢子虫分离株对雏鸡致病性比较研究

为研究不同禽源贝氏隐孢子虫对雏鸡的致病性差异,人工感染3日龄雏鸡鸡源、鸭源、鹌鹑源、白文鸟源贝氏隐孢子虫.结果发现,各感染组雏鸡潜隐期均为4d,显露期为11～18 d,均未发生死亡,但其增重均受影响.鸡源C.baileyi(林州株)、C.baileyi(郑州株)和鸭源C.bailey(郑州株)感染组雏鸡临床症状较为明显,发病率较高,分别为71.4％、85.7％和77.1％,剖检可见气管有较多粘液分泌物,法氏囊粘膜充血肿胀,平均增重较不感染对照组分别减少34.5、53.1和19.7 g；鹌鹑源C.bailey(武陟株)和白文鸟源C.bailey(郑州株)感染组雏鸡仅部分表现轻微临床症状,发病率分别为31.4％和28.6％,平均增重较不感染对照组分别减少39.7和43.1g.结果表明不同禽源C.bailey对雏鸡均有不同程度的致病性,以鸡源和鸭源C.baileyi致病性较强.     

保定地区奶牛源隐孢子虫的分离与基因鉴定

为了解保定地区奶牛源隐孢子虫感染情况,从当地3个奶牛场随机采集145份奶牛粪便经饱和蔗糖溶液漂浮后直接镜检和抗酸染色后镜检,阳性样品采用PCR方法扩增18S rRNA基因,同时将扩增出的目的片段进行克隆和序列分析,并将分离株与其他11个隐孢子虫参考株的18S rRNA序列进行比对和遗传距离比较。结果表明:有10份粪便样本为隐孢子虫阳性,感染率为6.9%(10/145),不同奶牛场、年龄段奶牛隐孢子虫感染率有显著性差异(P0.05)。10份粪便样品采用PCR方法扩增后有7份为18S rRNA基因阳性,扩增出的18S rRNA部分基因片段长528 bp。保定地区隐孢子虫分离株扩增的基因序列与牛源隐孢子虫18S rRNA基因序列(Gen Bank登录号为HQ179571)同源性最高,确定保定地区分离到的奶牛源隐孢子虫为牛隐孢子虫。     

广西2月龄内犊奶牛隐孢子虫感染情况调查

<正>自Tyzzer(1907)首次发现小鼠隐孢子虫(C.muris)以来,国内外学者对隐孢子虫病进行了大量研究,已在人、哺乳动物、禽类、爬行动物和鱼类等多种动物中发现15个种类的隐孢子虫感染[1-3],其中可感染牛的隐孢子虫种为安氏隐孢子虫(C.an-dersoni)、小球隐孢子虫(C.parvum)、牛隐孢子虫(C.bovis)、小鼠隐孢子虫(C.muris)、犬隐孢子虫(C.ca-     

河南地区鸭隐孢子虫病流行病学调查

隐孢子虫病是重要的人兽共患原虫病,广泛分布于世界各地.隐孢子虫(Crptosporidium)宿主种类广泛,可以寄生于240多种动物,隐孢子虫有效种目前已达到16个,另有40多个基因型.其中已确定的禽类有效种有3个,即贝氏隐孢子虫(C.baileyi)、火鸡隐孢子虫(C.meleagridis)和鸡隐孢子虫(C.galli),还可能存在新的有效种和基因型,至少存在有2个未命名种(分别寄生于鹌鹑和鸵鸟[1])和3个新基因型(从黑鸭分离到的鸭基因型和加拿大鹅分离到的两个鹅基因型)[2].……     

白术、微米白术和白术多糖对断奶仔猪生长性能和肠道形态及微生态区系的影响

试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。     

Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia

Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.     

Y‐chromosomal variation of local goat breeds of Turkey close to the domestication centre

Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.     

A brief history of the prevention of infectious diseases by immunisations

Infectious diseases have always been a terrible scourge for humans. The appearance of these plagues, as they were called without distinction, was generally connected to various conditions: asters, climatic changes or religious reasons. The concept of contagious, and then infectious, diseases came slowly. Variolation, i.e. transmission of ‘virulent’ matter to induce a natural disease and the immunity against it, was brought from Constantinople to England by Lady Montague, in 1721. This ‘variolation’ technique was also often performed in veterinary medicine against diseases like sheep-pox or pleuropneumonia. As ‘vaccination’ is the term generally accepted for ‘immunisation’, variolation can be the word designating such a technique. The second period of the history of immunisation began, in 1880, with the studies of Pasteur and his collaborators. A great number of bacterial vaccines were developed: dead, live but attenuated or only parts of pathogens. The viruses were produced in animals, then in eggs and at last, in tissue cultures. Second generation vaccines appeared with genetic engineering: recombinant vaccines, vector vaccines, nucleic acids vaccines, and markers vaccines, among others. These novel technologies can permit the development of new ones and improve the quality of the vaccines already existing.     

Micronutrient Content of Certain Tropical Conventional and Unconventional Feed Resources of Southern India

The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies.     

Bovine Mastitis in Selected Areas of Southern Ethiopia

A study on bovine mastitis, designed to determine the causal agents, prevalence of infection and impact of risk factors in three cattle breeds, was conducted in selected areas of southern Ethiopia. A total of 307 lactating and non-lactating cows, of which 162 were indigenous Zebu, 85 Jersey and 60 Holstein-Friesian, were examined by clinical examination and the California mastitis (CMT) test. Of these, 40.4% were positive by CMT and bacteriology for clinical or subclinical mastitis, with prevalence rates of 37.1% and 62.9%, respectively. Out of 1133 quarters examined, 212 (18.7%) were found to be infected, 83 (39.2%) clinically and 129 (60.8%) subclinically. The prevalence of mastitis was significantly higher in Holstein-Friesian than in indigenous Zebu, in non-lactating cows than in lactating cows, in the early lactation stage than in the mid-lactation stage, in cows with lesions and/or tick infestation on skin of udder and/or teats than in cows without this factor, and in the wet season than in the dry season. Mastitis increased with parity number (R = 0.9). Of 248 CMT and clinically positive udder quarter samples analysed microbiologically, 212 were culturally positive for known mastitis pathogens and 36 were negative. Of the 199 positive samples, Staphylococcus accounted for 39.2%, Streptococcus for 23.6%, coliforms for 14.1%, Micrococcus and Bacillus species for 8.0% each and Actinomyces or Arcanobacterium (Corynebacterium) for 7.0%. It was concluded that there was a high prevalence of clinical and subclinical mastitis, mainly caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli, in this study area.     

不同日粮氮水平对山羊氮代谢和微生物蛋白质合成的影响

本试验旨在探讨不同日粮氮水平对内蒙古白绒山羊氮代谢和微生物蛋白质合成(MCP)的影响,以达到提高氮利用率,减少环境污染和饲料资源的浪费。试验选用9只体况良好、体重为(43.83±2.95)kg,装有瘤胃瘘管的内蒙古白绒山羊,按体重随机分为3组,每组3只。日粮分为低氮7.5%、中氮10.5%、高氮13.5%3个氮水平。试验使用常规试验方法分析测定粪尿血液等生化指标,通过全收尿法和尿嘌呤衍生物法估测MCP。结果表明:随着氮水平增加,氨氮(NH3-N)浓度、氮摄入量、尿氮排出、尿中尿素氮(UUN)和MCP显著增加(P0.05),干物质采食量(DMI)显著降低(P0.01);沉积氮占总摄入氮的比例在中氮组最高,MCP在高氮组最高。综上所述,适当降低日粮氮水平可减少粪尿氮排放,提高反刍动物氮利用率。     

Microsporidial keratopathy in two dogs

A microsporidial keratopathy is described in two dogs. Both dogs presented with a unilateral stromal keratopathy characterized by multifocal coalescing opacities, and the diagnosis was made on histopathologic examination of keratectomy specimens. Transmission electron microscopy (TEM) on formalin‐fixed, paraffin‐embedded corneal tissue was performed in one dog, and the morphologic features were consistent with Nosema species infection. Both dogs were initially diagnosed and treated by superficial keratectomy. One dog received additional antifungal medication and underwent a penetrating keratoplasty following local recurrence two years later. No other systemic lesions attributable to the microsporidial infection were identified clinically. The clinical and diagnostic pathology findings, treatment, and follow‐up are discussed.     

优质肉鸡S3系体重与体尺性状指标的主成分分析

为研究地方优质肉鸡S3系体重和体尺性状之间的内在相关性,12周龄时选取同一饲养条件下的150只鸡(公母各半)进行体重和体尺性状等7个指标的测定。结果表明:S3系公鸡的体重、体斜长、龙骨长、胸角均显著高于母鸡(P0.05);公母鸡体重、体斜长、胸宽、胫围这4项指标变异系数较大,有较大选育空间;体重与体斜长、龙骨长、胸宽、胸角、胫围呈极显著正相关(P0.01),相关系数分别为0.57、0.52、0.51、0.49和0.47,与胸深、胫长呈显著相关(P0.05),相关系数分别为0.44和0.39;其他各体尺指标间也存在不同程度的正负相关性(P0.05或P0.01);体重和7个体尺性能指标综合成5个复合指标,累计贡献率达92.6355%,其中第l、2、3、4、5主成分分别解释总变异的36.5353%、22.8642%、13.3115%、10.4535%和8.4710%;各主成分的特征根分布较广,经统计计算入选的5个主成分所包含信息的侧重点各有相同,分别都在一定程度上反映了S3鸡的体型特征和生长发育规律,为今后选育出更加符合市场需求的屠宰加工优质型肉鸡指明了的方向。     

Efficacy of tropicamide,homatropine, cyclopentolate,atropine and hyoscine as mydriatics in Angora goats

Abstract

AIM: To document the efficacy of five commercially available mydriatics for their potential for diagnostic and therapeutic use in Angora goats.

METHODS: Over 8 weeks, the mydriatic effects of 1% tropicamide, 2% homatropine, 1% cyclopentolate, 1% atropine and 0.25% hyoscine were evaluated. Given as block treatments, drugs were applied randomly to one eye of 10 Angora goats, and the contralateral eye served as a control. Vertical and horizontal pupil diameters were measured to document onset ofeffect, time to reach a difference of 5 mm in the vertical/horizontal pupil diameter between eyes, time to maximum pupillary dilation, and duration of mydriatic action.

RESULTS: Onset of mydriasis for all drugs occurred within 15 minutes. Time to reach a difference of 5 mm in the vertical pupil diameter between eyes was shortest for 1% tropicamide and 0.25% hyoscine (0.5 h), then 2% homatropine and 1% atropine (0.75 h), and longest for 1% cyclopentolate (1.5 h). The maximum vertical pupillary dilation occurred earliest with 1% tropicamide and 1% atropine (2 h), followed by 0.25% hyoscine (3 h), 2% homatropine (4 h), and latest with 1% cyclopentolate (8 h). The duration of vertical dilation of the pupil was shortest with 1% tropicamide (6 h), then 2% homatropine (12 h), 1% cyclopentolate (12 h), 1% atropine (24 h), and longest for 0.25% hyoscine (96 h).

The time to reach maximum horizontal dilation of the pupil in treated eyes was shortest with 1% cyclopentolate (1 h), followed by 1% tropicamide (1.5 h), 0.25% hyoscine (3 h), 2% homatropine (3.5 h), and 1% atropine (4 h). The duration of horizontal pupil dilation was shortest with 1% tropicamide (4.5 h), and longest with 0.25% hyoscine (48 h).

CONCLUSION: All five mydriatics induced clinical dilation. Tropicamide (1%) had the shortest duration of effect, but gave incomplete dilation. Good dilation was achieved with 1% cyclopentolate and 2% homatropine, but took too long to reach maximum dilation for routine mydriasis. The largest vertical dilation of the pupil was achieved with 1% atropine and 0.25% hyoscine, but pupils remained dilated for more than 24 h.

CLINICAL RELEVANCE: For routine mydriasis in goats, it is recommended that 1% tropicamide be used, though there may be incomplete dilation. For a longer duration of mydriasis, such as in the treatment of anterior uveitis, 1% atropine or 0.25% hyoscine would be the drugs of choice.