西藏牦牛源枯草芽孢杆菌的分离培养及鉴定

从西藏健康牦牛的新鲜粪便中分离、纯化枯草芽孢杆菌,并筛选出耐受性最好的菌株,并进行形态学观察、生化试验、PCR扩增、电泳检测、16s rRNA序列测定、构建进化树并进行同源性分析,对通过鉴定的枯草芽孢杆菌进行耐肠液、耐胆盐、耐高温等耐受性试验.结果显示:分离出疑似枯草芽孢杆菌4株,革兰氏染色为阳性;芽孢染色液染色菌体为...     

鸡约氏乳杆菌的分离鉴定及进化分析

从3日龄健康小鸡肠道中分离到1株乳杆菌,用PCR方法从分离菌株扩增16S rRNA基因,获得大小为1340 bp的DNA片段,该片段的核酸序列已提交GenBank,登录号为EU290749。将分离株的16S rRNA基因核苷酸序列与GenBank上其它乳杆菌进行同源性分析并建立进化树。结果表明,分离株的16S rRNA基因核苷酸序列与NCBI公布的鼠约氏乳杆菌分离株(AB295648)的同源性为99.6%,因此该分离菌株被鉴定为鸡约氏乳杆菌(chicken Lactobacillus johnsonii),命名为HN/0711。     

猪源枯草芽孢杆菌的分离鉴定及其生物特性研究

为研究和开发动物新型微生态制剂,改变抗生素滥用、药物残留等现状。通过热处理从健康猪肠道内容物、粪便、土壤中分离芽孢杆菌,结合形态特征、生理生化特性及16S rRNA序列分析进行鉴定;对菌株耐胆盐性、酸性耐受性及产酶情况等特性进行研究。结果表明,分离到11株芽孢杆菌,经细菌形态学、生理生化特征和16S rRNA序列分析的结果显示,3株为枯草芽孢杆菌;特性研究结果证实菌株SR-096具有较好的耐胆盐性、酸性耐受性和产脂肪酶、淀粉酶、蛋白酶和纤维素酶的能力。说明枯草芽孢杆菌SR-096具有较强的抗逆性和潜在的益生性能,为研制微生态制剂提供了优良的菌种参考。     

一株鸭源枯草芽胞杆菌的分离鉴定及药敏试验

为了鉴定从患鸭疫里默氏杆菌病鸭脑组织中分离到的混合感染细菌,采用常规方法观察分离菌株的菌落形态、革兰染色特征、生化反应特点,采用细菌16S rDNA PCR及序列分析方法对分离菌株进行分子生物学鉴定。结果表明,分离菌株在胰蛋白大豆肉汤培养基上形成边缘不规则,凸起,表面灰白色,粗糙的大菌落;镜检呈革兰阳性杆菌,能够产生芽胞;在兔血平板上生长时出现明显的β溶血环;生化反应特点与枯草芽胞杆菌模式株基本相符,但略有不同;其16S rDNA序列与GenBank中已经报道的枯草芽胞杆菌16S rDNA序列的相似性达到99%,最终确定分离菌株为枯草芽胞杆菌;分离菌株对红霉素、万古霉素、阿莫西林等抗菌药物敏感,对克林霉素和多西环素中等敏感,仅对头孢噻肟耐药。研究结果为临床治疗由鸭疫里默氏杆菌和枯草芽胞杆菌引起的混合感染提供了科学用药方案。     

水貂肠道枯草芽孢杆菌的分离及16S rRNA鉴定

为了获得水貂的有益菌种,及研制毛皮动物益生菌制剂提供基础,并为进一步深入研究枯草芽孢杆菌提供参考,从健康水貂的肠道中分离纯化出1株枯草芽孢杆菌,编号为 P2,通过形态学观察、生理生化试验以及16S rRNA 序列分析及同源性比较进行综合鉴定,结果显示菌株 P2与菌株 Bacillus subtilis CICC 10023（GU980947.1）亲缘关系最近,同源性达到了99.9%,因此鉴定 P2菌株为枯草芽孢杆菌。     

母猪粪便中乳酸菌菌株体外益生特性初步研究

试验从健康母猪粪便中分离乳酸菌,并对其益生特性进行初步研究。通过形态和生理生化鉴定,再利用16S rDNA序列同源性分析的方法进行16S rDNA基因扩增与测序,并将测序结果与GenBank中菌株的16S rDNA基因序列进行同源性比较分析,得到1株链球菌,2株约氏乳杆菌,4株乳酸片球菌。体外益生特性结果表明,各菌株在pH 2.0~3.0,胆盐浓度0.1%~0.3%均有较强的耐受能力,各菌株对大肠埃希氏菌和单核细胞增多症李氏杆菌均有较好的抑制作用。     

副猪嗜血杆菌的分离鉴定及16S rRNA序列分析

从云南某规模化养猪场病猪肺脏分离到1株革兰氏阴性小杆菌,经细菌生化鉴定、PCR鉴定和16S rRNA序列比对鉴定为副猪嗜血杆菌。抗生素药物敏感试验结果表明,分离菌株对四环素、红霉素、氯霉素、头孢噻吩高敏;对庆大霉素、氧氟沙星、诺氟沙星中敏;对磺胺甲唑耐药。16S rRNA分析结果表明,该分离株与GenBank中的Hps参考株AB078973(基因登录号)同源性为100%,将分离菌株鉴定为副猪嗜血杆菌。16S rRNA遗传进化关系表明,分离株与副猪嗜血杆菌3株血清5型参考株AB078972、AB078973、AB078974的16S rRNA序列位于一个分支上,遗传进化关系最近,它们之间的核苷酸同源性在99.0%~99.4%之间,初步鉴定为血清5型副猪嗜血杆菌,致病性试验结果表明,分离菌株对小白鼠有强致病性,命名为YN-1株。     

牦牛源蜡样芽胞杆菌的分离鉴定及耐药性分析

为了研究牦牛源蜡样芽胞杆菌致病性和耐药性,本实验对其进行了细菌形态学观察、16S rDNA PCR和序列比对、动物回归试验及药敏分析。结果显示：56份牦牛奶样,分离出135株细菌,对分离菌进行16S rDNA PCR并测序,其中有42株为蜡样芽孢杆菌,蜡样芽胞杆菌检出率为75%(42/56),占总检出细菌的31.11%(42/135);其中分离菌株B2019 16S rDNA PCR测序结果与Gen Bank中蜡样芽胞杆菌的同源性为88.6%～99.3%,运用Mega7.0将分离菌株B2019与6株参考菌株的16S rDNA序列进行同源性比对分析,并构建系统进化树;在动物回归试验中小鼠腹泻,剖检小鼠发现肝脏肿大,肺脏边缘有出血;药敏试验结果显示,分离菌株对麦迪霉素、克林霉素、红霉素、庆大霉素、卡那霉素、丁胺卡那霉素、新霉素、万古霉素、氧氟沙星、环丙沙星、呋喃唑酮和米诺环素等13种抗生素表现出高度敏感,对多粘菌素B表现为中介,对其他16种抗生素均表现为耐药。试验结果表明该分离菌株是一株具有较强致病性的蜡样芽胞杆菌。     

奶牛阴道分泌物中枯草芽孢杆菌的分离鉴定及抑菌效果分析

从奶牛阴道分泌物中分离出一株细菌,经过菌落形态特征、生理生化鉴定初步判定为芽孢杆菌属;根据16S rRNA基因序列及其同源性分析结果,证实分离得到的Y4菌株是枯草芽孢杆菌,与GenBank上的枯草芽孢杆菌16S rRNA基因序列同源性为99.6%。对此菌株进行抑菌试验,发现该菌株对大肠杆菌、金黄色葡萄球菌和链球菌有较好的抑菌效果。该研究为将来研制益生菌制剂防治奶牛生殖道疾病提供了物质基础。     

副猪嗜血杆菌广东流行株的分离鉴定与基因分析

本研究从广东省各个地区送检病料中成功分离鉴定了4株副猪嗜血杆菌,并且针对副猪嗜血杆菌16S rRNA基因特异性进行PCR检测和基因测序同源性分析,通过GenBank联机比对分析,所分离的菌株与国内外菌株16S rRNA序列同源性在98.2%以上,分离菌株之间同源性在99.6%~100%之间,证明了所暴发的菌株是副猪嗜血杆菌。     

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.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

Internal Parasites and Haematological Values in Cattle Slaughtered in Buea Subdivision of Cameroon

Tropical Animal Health and Production -     

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.     

Chemotaxis of macrophage in inflammation

Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.     

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