旋毛虫T668基因转录及表达的时空特性鉴定

采用RT-PCR和免疫荧光染色技术,分别检测T668基因mRNA及蛋白在旋毛虫不同发育时期中的时空表达情况。RT-PCR结果显示,该基因从旋毛虫新生幼虫出生即开始转录,随着日龄的增加,T668基因mRNA逐渐减少,在感染后11～14 d T668基因的转录趋于停止。免疫荧光染色结果表明,在感染后1d,旋毛虫虫体即有T668蛋白表达;感染后6～13d,T668蛋白分泌到肿大的肌细胞核上;14～20d,肿大的肌细胞核上基本观察不到T668蛋白,但整个虫体仍有很强的蛋白表达。这提示在旋毛虫的寄生过程中,T668基因的转录及表达呈一定的时空特点,这可能对旋毛虫包囊形成有着重要作用。     

PCR检测感染早期小鼠血液中旋毛虫幼虫的研究

为了研究PCR检测感染小鼠血液中旋毛虫DNA的敏感性,应用旋毛虫1.6 kb重复序列为扩增靶序列对旋毛虫(T1)、乡土旋毛虫(T2)、布氏旋毛虫(T3)、伪旋毛虫(T4)和南方旋毛虫(T7)肌幼虫DNA进行PCR扩增,并检测小鼠感染20、100、300条T1肌幼虫后不同时间的外周血.结果表明,T1、T4和T7肌幼虫可扩增出特异性目的条带(510 bp),而T2和T3无扩增产物;1、0.04和0.02条T1、T4和T7肌幼虫均能扩增到清晰的目的条带(510 bp).20条幼虫感染小鼠后5 d～6 d,PCR阳性率均为7.69%;100条幼虫感染小鼠后5 d～12 d可检出旋毛虫DNA,其中感染后5 d～7 d的阳性率分别为30.77%、38.46%及30.77%;300条幼虫感染小鼠后5 d～15 d可检出旋毛虫DNA,感染后7 d的阳性率为61.54%,感染后6 d与8 d～10 d的阳性率均为53.85%. 3组旋毛虫感染小鼠PCR阳性率间的差异有统计学意义(p<0.01),PCR阳性率随感染剂量的增加而升高(p<0.01),100条与300条感染小鼠感染后不同时间的PCR阳性率与检测时间有相关性(p<0.01).以上实验结果表明PCR检测感染小鼠血液中旋毛虫DNA的敏感性与感染程度和检测时间有关,对感染早期旋毛虫抗体阴性宿主有一定诊断价值.     

牛旋毛虫病肌组织病理形态学及包囊形成的观察

用旋毛虫对3头牛成功地进行了实验性人工感染,并用光学显微镜和电子显微镜系统观察了牛肌旋毛虫的形态学变化、肌组织的病理变化及旋毛虫包囊的形成。经口感染旋毛虫后第7d,牛肌组织中出现旋毛虫幼虫,呈杆状,位于肌纤维之间。感染后11d,幼虫进入肌纤维之中,呈杆状。感染后19d,幼虫的一端出现弯曲,并开始包囊的形成,感染后27d,幼虫蟠曲,100d以后,形成完整包囊,其形状为梭形或卵圆形。电镜下观察,包囊壁分为内、外两层,外层由胶原微纤维组成,来自增生的内芽肿;内层由不定形基质、不能显示周期性横纹的胶原微纤维和小泡组成,来自受侵肌纤维。     

猪旋毛虫和囊虫合并感染病例

1988年4月13日河南省中牟县送入我厂190头猪,宰后检出囊虫12头,旋毛虫6头,其中1头属旋毛虫和囊虫合并感染。检出率为8.95%。合并感染猪只40平方厘米内有4—6个囊虫虫体,后腿、前腿、颈背肌肉均有寄生;旋毛虫镜检24个膈角肉片     

斯氏艾美耳球虫四川株生物学特性研究

为了解斯氏艾美耳球虫四川株致病性、免疫原性等生物学特性,为后续免疫和防控等研究提供参考。将45只36日龄幼兔随机分为9组,第1组为空白对照组,第2组～第5组为致病性试验组,于50日龄每只兔分别经口感染1×10~4、5×10~4、1×10~5、2×10~5个孢子化卵囊;第6组～第9组为免疫原性试验组,于36日龄每只兔分别口服接种5×10~2、5×10~3、1×10~4、5×10~4个孢子化卵囊,14 d后给予2×10~5个孢子化卵囊的感染量。测定最小潜隐期和排卵高峰并以临床症状、卵囊排出量、相对增重、肝指数、肝脏病理变化作为评价指标。分离培养斯氏艾美耳球虫四川株卵囊,测定其最短孢子化时间及卵囊大小。结果表明,斯氏艾美耳球虫四川株最小潜隐期是13 d,排卵高峰在感染后22 d～23 d,孢子化卵囊的平均大小为36.2μm(28.2μm～38.1μm)×19.9μm(17.80μm～22.3μm),卵囊指数为1.81,27℃恒温培养最短孢子化时间为41 h。致病性试验组与空白对照组肝指数、平均增重、相对增重率差异显著(P0.05);免疫原性试验组与攻毒对照组肝指数、平均增重、相对增重率差异不显著(P0.05)。斯氏艾美耳球虫四川株致病性较强,低剂量感染就能显著降低增重并引起明显病变,但其免疫原性较差。     

小鼠实验感染不同种旋毛虫后肉汁抗体水平的研究

为了观察小鼠感染不同种旋毛虫后肉汁抗体水平的变化及其与血清抗体水平的相关性，将200只昆明小鼠随机分成4组（每组50只），每只分别感染300条乡土旋毛虫（T2）、布氏旋毛虫（T3）、伪旋毛虫（T4）及纳氏旋毛虫（T7）幼虫，感染后2～6周每组每周随机剖杀10只小鼠，收集血清及肉汁，用旋毛虫肌幼虫ES抗原ELISA检测血清及肉汁中抗体水平；另取40只昆明小鼠随机分成4组（每组10只），每只分别感染500条T2、T3、T4、T7幼虫，感染后6周剖杀，制备肉样，用ELISA检测4℃及-20℃保存不同时间后的肉汁抗旋毛虫抗体动态。T2、T3、T4、T7感染小鼠后的肉汁抗体水平和变化趋势相似，均是在感染后第3周检测出肉汁特异性抗体，抗体阳性率分别为60％、30％、30％、30％，至感染后第4周肉汁抗体阳性率均升至100％。4组小鼠感染后2～6周的肉汁与血清抗体水平均具有相关性。T2、T3、T4、T7感染小鼠肌肉4℃保存7d与1d的肉汁抗体水平相比均无显著性差异；所有的感染小鼠肌肉-20℃保存2个月的肉汁抗体水平与保存1个月的相比均无显著性差异；虽然保存3个月的肉汁抗体水平与保存1个月的相比均已有显著性差异，但抗体阳性率仍均为100％并持续至实验结束时的4个月保存期。结果表明，肉汁中抗旋毛虫抗体的检测可用于新鲜肉、冷藏肉及冷冻肉中乡土旋毛虫、布氏旋毛虫、伪旋毛虫、纳氏旋毛虫捡疫的初筛。     

胶体金法检测猪旋毛虫病的敏感性研究

将3组试验仔猪分别人工感染旋毛虫幼虫1000条／头、5000条／头和25000条／头,感染后每7d抽血1次,采集全血和血清;感染后46d,将试验猪全部剖杀,取全血、血清和膈肌肉待检。所有样品均用胶体金试纸检测猪旋毛虫抗体和抗原,膈肌肉样品并需用显微镜检查有无旋毛虫包囊。结果表明,最早可于感染后21d,在低感染量试验组的1头仔猪的血清中检出阳性抗体;血清样品的抗体检出率高于全血样品;在血液和血清中未检出虫体抗原;抗体试纸条的检测结果较为准确可靠,而且敏感性明显强于抗原试纸条。建议在检疫检验工作中使用抗体试纸条检测血清或肌肉样品。     

犬吉氏巴贝斯虫病的诊断与治疗

犬吉氏巴贝斯虫,虫体小,直径约1～3.3μm,呈环形或椭圆形,寄生于红细胞内可引起溶血性贫血和黄疸等症,患犬感染初期,血液涂片经瑞士染色或吉姆萨染色可见虫体呈深紫色。本病常呈慢性经过,初期持续发热3～5 d,随后恢复正常,呈现不规则回归热。笔者于2012年5月诊治一例犬吉氏巴     

旋毛虫幼虫“感染力”和“肉眼可见度”的测定——对食肉旋毛虫“目检法”的兽医卫生评价

食肉旋毛虫的检验是兽医卫生检验工作中的一个重要环节。我国现行法定的检验方法是“压片镜检法”。 苏联学者教授指出:“旋毛虫幼虫于感染后第40～50天,在宿主肌组织中完成包囊的建造过程。之后,便具有对其它动物和人的侵袭能力”。还指出:“在沿肌纤维新剪开的肌肉上,若视力良好,则用肉眼亦能发现具有包囊的虫体,呈乳白色、微尘状”。 国内不少单位经实验观察与调查研究,结果表明:“旋毛虫自感染猪体后21天至7     

旋毛虫致小鼠心肌损伤的试验

本研究旨在对重度感染旋毛虫后小鼠心肌损伤的研究。通过对小鼠接种旋毛虫,1 000条/只,分别于不同时间对血清中心肌损伤标志物H-FABP、CK-MB和cTnT进行动态检测,以及心肌组织病理形态学变化进行观察。结果显示,H-FABP比CK-MB、cTnT敏感性高(P<0.05),出现及达到高峰的时间最早;感染旋毛虫后19d～24d,心肌组织损伤最为严重。H-FABP可用于早期旋毛虫病患者心肌损伤的诊断,具有较高的敏感性和特异性。     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ

An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein     

Evaluation of the Microbiological Status of Milk and Various Structures in Mammary Glands from Naturally Infected Dairy Cows

A knowledge of the microbiological status of milk and of the different structures in the mammary glands has great importance in elucidating the pathogenesis of mammary gland infections. The objective of this study was to evaluate the microbiological status of various structures in the mammary glands from naturally infected dairy cows following slaughter. A total of 94 samples of milk, 184 samples of mammary parenchyma, 168 samples of gland cisterns, and 168 samples of teat cisterns were collected for microbiological examination. Microorganisms were detected in 59.9% of all samples, 67.0% of the milk samples, 70.1% of the mammary parenchymas, 55.9% of the gland cisterns and 48.8% of the teat cistern samples. When all samples were considered, coagulase-negative Staphylococcus were the most prevalent (35.7%) followed by coagulase-positive Staphylococcus (12.2%), Corynebacterium bovis (2.4%), Prototheca sp. (1.9%), and Streptococcus dysgalactiae (1.5%). There was a significantly higher occurrence of microorganisms in the milk and mammary parenchyma compared to the gland cisterns and teat cisterns. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development and survival of free-living stages of gastrointestinal nematodes of sheep and goats on pasture in the Nigerian derived savanna

Four grass plots were sequentially contaminated with goat faeces containing known numbers of unembryonated eggs of predominantly Haemonchus contortus and Trichostrongylus spp. between October 1982 and April 1983. Four other plots were similarly contaminated with sheep faeces between February and May 1987. An additional plot was repeatedly contaminated with sheep faeces from February to April 1987. Populations of free-living stages in faeces and of infective larvae (L₃) in the herbage were subsequently monitored until the end of April and June of 1983 and 1987 respectively. During February and May 1987 two control cultures of sheep faeces were incubated in the laboratory at 25°C–30°C and at a constant temperature of 50°C and the free-living development was also monitored. L₃ developed very readily in the faeces cultured at 25°C–30°C and in those spread on a grass plot in October, at the end of the wet season, but developed less on the plot contaminated in May at the start of the wet season. Worm eggs in faeces deposited on plots during the hot dry season (December to April) or incubated at 50°C died and disintegrated after 24–48 h exposure to the high environmental temperatures. The results indicate that it is unlikely that gastrointestinal nematodes of sheep and goats can develop or survive on open pasture during the dry season in the Nigerian derived savanna zone.