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1.
提取鸡致病性大肠杆菌分离株O1，O2和O78的基因组DNA作为模板，用TD－PCR技术从其中分别扩增出了0．55kb的I型菌毛细菌基因（piliA)，将扩增得到的piliA基因片段，TA克隆的方法分别克隆进pGEM-T载体中，转化至受体菌JM109中，用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法，得到含阳性重组子的菌株，提取质粒用PstI单酶切及NcoI和PstI双酶切鉴定，结果证实，所构建的克隆质粒中均含有相应piliA基因，经DNA序列分析，其结果基因阅读框架大小为549bp，但其中O1菌株I型菌毛基因在第72位发生突变，有6个碱基插入，经DNA Star核酸分析软件分析，此3菌株的I型菌毛基因的同源性为89．9％－92％。  相似文献   

2.
提取鸡致病性大肠杆菌分离株O1、O2和O78的基因组DNA作为模板,用TD-PCR技术从其中分别扩增出了0.55kb的Ⅰ型菌毛结构基因(piliA).将扩增得到的piliA基因片段,用TA克隆的方法分别克隆进pGEM-T载体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得到含阳性重组子的菌株,提取质粒用PstI单酶切及NcoI和PstI双酶切鉴定,结果证实,所构建的克隆质粒中均含有相应piliA基因.经DNA序列分析,其结构基因阅读框架大小为549 bp,但其中O1菌株Ⅰ型菌毛基因在第72位发生突变,有6个碱基插入.经DNA Star核酸分析软件分析,此3菌株的Ⅰ型菌毛基因的同源性为89.9%～92%.  相似文献   

3.
提取鸡致病性大肠杆菌分离株O1、O2和O78的基因组DNA作为模板,用TD-PCR技术从其中分别扩增出了0.55kb的I型菌毛结构基因(piliA)。将扩增得到的piliA基因片段,用TA克隆的方法分别克隆进pGEM-T载体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得到含阳性重组子的菌株,提取质粒用PstI单酶切及NcoI和PstI双酶切鉴定,结果证实,所构建的克隆质粒中均含有相应piliA基因。经DNA序列分析,其结构基因阅读框架大小为549bp,但其中O1菌株I型菌毛基因在第72位发生突变,有6个碱基插入。经DNAStar核酸分析软件分析,此3菌株的I型菌毛基因的同源性为89.9%~92%。  相似文献   

4.
提取鸡致病性大肠杆菌分离株O1、O2和O78的基因组DNA作模板,用TD-PCR技术从其中分别扩增出0.55 KB的I型菌毛结构基因(piliA).将扩增得到的piliA基因片段,用TA克隆的方法分别克隆进pGEM-T栽体中,转化至受体菌JM109中,用Amp/IPTG/X-gal琼脂平板蓝白菌落筛选法,得到舍阳性重组子的菌株,提取质粒用PstI单酶切及NcoI和PstI双酶切鉴定,结果证实,所构建的克隆质粒中均含有相应piliA基因.经DNA序列分析,其结构基因阅读框架大小为549 bp,但其中O1菌株Ⅰ型菌毛基因在第72位发生突变,有6个碱基插入.经DNAStar核酸分析软件分析,3个基因同源性为89.9%～92.0%.  相似文献   

5.
从山东省各地分离到107株鸡致病性大肠杆菌，选择了其中2株高致病性大肠杆菌，血清型分别为O78和OM，经增菌培养后提取菌毛制备为单价菌毛油乳苗，分别接种于1日龄和14日龄雏鸡，于4周龄攻毒。结果二者之间有一定的交叉保护作用。根据Genbank收录的人源大肠杆菌I型菌毛FimA基因和P型菌毛papA基因序列，分别设计了2对引物。通过PCR对上述2株大肠杆菌进行扩增，结果只有FimA的一对引物扩增出相应的条带，经测序证明为FimA基因。papA基因的引物未扩出任何条带，证明这2株大肠杆菌表达I型菌毛。通过对2株大肠杆菌结构基因FimA进行分析，发现二者具有高度的同源性。本研究的目的是探讨菌毛亚单位之间的交叉保护性与其菌毛的结构基因之间是否存在相关性。  相似文献   

6.
研究选择从四川规模化鸡场分离鉴定的5株优势血清型鸡源致病性大肠杆菌代表株(O89,O119,O141,O127)的1型菌毛pilA基因的PCR扩增片段分别定向克隆到pUC18质粒的多克隆位点,并转化到DH5 α株大肠杆菌载体菌中,对目的基因的序列测定结果表明：5个克隆片段均含有1型菌毛pilA基因的全序列,全长459bp,编码信号肽和结构蛋白。用生物软件(DNASTAR)对9株大肠杆菌1型菌毛pilA基因序列、氨基酸序列、菌毛蛋白质二级结构预测及抗原性进行了分析比较,结果显示：鸡源致病性大肠杆菌1型菌毛间具有同源性,1型菌毛间存在一定的相同抗原位点。  相似文献   

7.
从禽源大肠杆菌037(O78)、166(O78)、120(O18)分离株和猪源大肠杆菌107/86分离株分别提取基因组DNA,并以此为聚合酶链反应(polymerasechainreaction,PCR)的模板,扩增上述分离株的1型菌毛主要亚单位结构基因pilA,得到了大小约570bp的扩增产物,将此扩增产物克隆于T载体,通过内切酶酶切分析得到4个阳性重组质粒;对上述4个大肠杆菌分离株的pilA基因进行序列测定,通过其编码的主要菌毛亚单位FimA蛋白氨基酸的序列比较发现:3个禽源株间FimA的同源性为94.3%至99.0%;禽源株和猪源株间FimA的同源性为89.6%至91.1%。在pilA开放性阅读框所编码的FimA182个氨基酸序列中,禽源大肠杆菌O78血清型的2个分离株037株和166株间只有2个氨基酸不同,其同源性为99.0%。  相似文献   

8.
从禽源大肠杆菌037(O78)、166(O78)、120(O18)分离株和猪源大肠杆菌107／86分离株分别提取基因组DNA，并以此为聚合酶链反应(Polymerase chain reaction，PCR)的模板，扩增上述分离株的1型菌毛主要亚单位结构基因pilA，通过其编码的主要菌毛亚单位FimA蛋白氨基酸的序列比较发现：3个禽源株间FimA的同源性为94．3％至99．0％；禽源株和猪源株间FimA的同源性为89．6％至91．1％。通过对重组大肠杆菌的菌体裂解物的SDS-PAGE电泳分析及Western blot分析，禽源大肠杆菌O78 037株、O18 120株出现了一致的强反应，O78 166株反应较弱，而猪源大肠杆菌107／86株反应最弱。这些结果表明：禽源大肠杆菌与猪源大肠杆菌1型菌毛间存在抗原多样性，这种多样性甚至出现在禽病原性大肠杆菌同一血清型的2个不同分离株之间，如O78 037株O78 166株之间，尽管其FimA氨基酸的同源性很高，为99．0％。  相似文献   

9.
用限制性内切酶SacI和BamHI双酶切克隆有鸡致病性大肠杆菌分离株O1、O2和O78的piliA基因的质粒pTFimO1、pTFimO2和pTFimO78,分别回收从此3质粒上切下的545bp的piliA基因片段,再将载体pET-28a(+)用SacI和BamHI双酶切,最后将pET-28a(+)DNA片段与545bp的piliA DNA片段进行连接,转化至受体菌BL21(DE3)中,经SacI和BamHI酶切反应鉴定重组子,得到了理想重组子质粒pETFimO1、pETFimO2和pETFimO78.重组菌株BL21(DE3) (pETFimO1)、BL21(DE3)(pETFimO2)和BL21(DE3)(pETFimO78)经IPTG诱导后,其表达产物经SDS-PAGE检测,结果表明重组菌株可以良好地表达鸡致病性大肠杆菌Ⅰ型菌毛蛋白.  相似文献   

10.
用限制性内切酶SacI和BamHI双酶切克隆有鸡致病性大肠杆菌分离株O1、O2和O78的piliA基因的质粒pTFimO1、pTFimO2和pTFimO78，分别回收从此3质粒上切下的545bp的piliA基因片段，再将载体pET-28a( )用SacI和BamHI双酶切，最后将pET-28a( )DNA片段与545bp的piliA DNA片段进行连接，转化至受体菌BL21（DE3）中，经SacI和BamHI酶切反应鉴定重组子，得到了理想重组子质粒pETFimO1、pETFimO2和pETFimO78。重组菌株BL21（DE3），（pETFimO1)、BL21（DE3）（pETFimO2)和BL21（DE3）（pETFimO78)经IPTG诱导后，其表达产物经SDS－PAGE检测，结果表明重组菌株可以良好地表达鸡致病性大肠杆菌I型菌毛蛋白。  相似文献   

11.
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.  相似文献   

12.
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.  相似文献   

13.
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.  相似文献   

14.
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.  相似文献   

15.
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.  相似文献   

16.
Comparison of different tests to diagnose feline infectious peritonitis   总被引：1，自引：0，他引：1  
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17.
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.  相似文献   

18.
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  相似文献   

19.
正We are pleased to publish the second special issue on avian brood parasitism and to be responsible guest editors for the two special issues of Chinese Birds (Vol. 3, No. 4, 2012 and Vol. 4, No. 1, 2013), entitled "Avian Brood Parasitism - A Growing Research Area in Behavioral Ecology". The first issue was published in December 2012. The goal of the two special issues is to publish accumulated knowledge and some of the recent developments in  相似文献   

20.
Tropical Animal Health and Production -  相似文献   

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