猫爪草内生真菌的分离、鉴定及抑菌活性的初步研究

植物内生真菌是一类具有开发潜力的新型微生物资源。为了从药用植物猫爪草中分离有生物活性的内生真菌,试验采用常规组织块分离法分离内生真菌,形态学及分子鉴定结合进行菌株分类,圆纸片法和对峙培养法检测抑菌活性。结果表明:从栽培猫爪草的根、茎、叶中分离纯化获得23株内生真菌,形态学初步鉴定为7个属;检测到有8株内生真菌对病原细菌金黄色葡萄球菌和病原真菌核盘菌、灰葡萄孢菌都有抑制作用。同属镰刀菌(Fusarium sp.)的菌株R1、L6、L11,刺盘孢菌(Colletotrichum sp.)P2及链格孢菌(Alternaria sp.)L2菌株具有开发生物防治药物的潜力,值得进一步深入研究。     

紫花苜蓿内生细菌的拮抗作用及优良拮抗菌株的鉴定和生物功能

从紫花苜蓿(Medicago sativa)茎部及根部分离内生细菌,采用对峙培养法测定内生细菌对紫花苜蓿根腐病菌厚垣镰孢菌(Fusarium chlamydosporum)、辣椒根腐病菌尖孢镰刀菌(Fusarium oxysporum)和茄镰孢菌(Fusarium solani)、黄瓜枯萎病菌(Fusarium oxysporum)、马铃薯枯萎病菌(Fusarium oxysporum)和核桃枝枯病菌燕麦镰刀菌(Fusarium avenaceum)和锐顶镰刀菌(Fusarium acuminatum)的拮抗作用,测定优良拮抗菌株的固氮、溶磷(无机磷)和分泌生长素的能力,并对所筛选的优良拮抗内生细菌进行形态学特征和16S rDNA基因序列分析鉴定。结果表明,从紫花苜蓿茎部及根部共分离得到的80株内生细菌中,39株对紫花苜蓿根腐病菌的抑菌率高于50%,9株抑菌率在61%以上,分别为MS-43 (63.96%)、MS-40 (63.75%)、MS-46 (63.54%)、MS-2 (63.13%)、MS-31 (62.29%)、MS-52 (62.08%)、MS-80 (61.88%)、MS-55 (61.46%)和MS-33 (61.25%);MS-33、MS-40、MS-43、MS-46和MS-52对其他6株镰刀菌(Fusarium spp.)的抑菌率均高于60%,MS-43的平均抑菌率达66.11%,具有固氮能力和分泌生长素能力;通过培养性状、形态特征和16S rDNA序列相似性分析,菌株MS-43属于类芽孢杆菌属(Paenibacillus)。     

采用EF-1α序列分析法对苜蓿根腐病病原菌——锐顶镰刀菌的鉴定

苜蓿根腐病对苜蓿生长和产量造成严重影响,为了解引起根腐病病原真菌的种类,本文对从陕西省定边县牧草草场紫花苜蓿根部分离得到的根腐病病原菌进行形态学观察、EF-1α序列分析鉴定及接种试验。结果表明:通过对病原菌在PDA培养基中的分离、纯化得到的单菌落形态学观察及显微镜下菌丝、孢子的观察,该病原菌属于引起苜蓿根腐病的镰刀菌属(Fusarium);用CTAB法提取病原菌基因组DNA,并对EF-1α序列进行PCR扩增、回收纯化、克隆测序,将测序结果进行Blast比对,构建系统发育树,分析与Genbank已知近缘种属亲缘关系,结果显示与锐顶镰刀菌(F.acuminatum)亲缘关系最近,可信度达99%;最后通过根部接种试验,接种苜蓿根部发病症状与田间根腐病发病症状一致,鉴定结果显示本试验研究的引起紫花苜蓿根腐病的病原菌为镰刀菌属锐顶镰刀菌(F.acuminatum)。     

紫花苜蓿内生细菌的拮抗作用及优良拮抗菌株的鉴定和生物功能

从紫花苜蓿(Medicago sativa)茎部及根部分离内生细菌,采用对峙培养法测定内生细菌对紫花苜蓿根腐病菌厚垣镰孢菌(Fusarium chlamydosporum)、辣椒根腐病菌尖孢镰刀菌(Fusarium oxysporum)和茄镰孢菌(Fusarium solani)、黄瓜枯萎病菌(Fusarium oxysporum)、马铃薯枯萎病菌(Fusarium oxysporum)和核桃枝枯病菌燕麦镰刀菌(Fusarium avenaceum)和锐顶镰刀菌(Fusarium acuminatum)的拮抗作用,测定优良拮抗菌株的固氮、溶磷(无机磷)和分泌生长素的能力,并对所筛选的优良拮抗内生细菌进行形态学特征和16S?rDNA基因序列分析鉴定.结果表明,从紫花苜蓿茎部及根部共分离得到的80株内生细菌中,39株对紫花苜蓿根腐病菌的抑菌率高于50％,9株抑菌率在61％以上,分别为MS-43?(63.96％)、MS-40?(63.75％)、MS-46?(63.54％)、MS-2?(63.13％)、MS-31?(62.29％)、MS-52?(62.08％)、MS-80?(61.88％)、MS-55?(61.46％)和MS-33?(61.25％);MS-33、MS-40、MS-43、MS-46和MS-52对其他6株镰刀菌(Fusarium spp.)的抑菌率均高于60％,MS-43的平均抑菌率达66.11％,具有固氮能力和分泌生长素能力;通过培养性状、形态特征和16S?rDNA序列相似性分析,菌株MS-43属于类芽孢杆菌属(Paenibacillus).     

采用EF-1α序列分析法对苜蓿根腐病病原菌——锐顶镰刀菌的鉴定

苜蓿根腐病对苜蓿生长和产量造成严重影响,为了解引起根腐病病原真菌的种类,本文对从陕西省定边县牧草草场紫花苜蓿根部分离得到的根腐病病原菌进行形态学观察、EF-1α序列分析鉴定及接种试验。结果表明:通过对病原菌在PDA培养基中的分离、纯化得到的单菌落形态学观察及显微镜下菌丝、孢子的观察,该病原菌属于引起苜蓿根腐病的镰刀菌属(Fusarium);用CTAB法提取病原菌基因组DNA,并对EF-1α序列进行PCR扩增、回收纯化、克隆测序,将测序结果进行Blast比对,构建系统发育树,分析与Genbank已知近缘种属亲缘关系,结果显示与锐顶镰刀菌(F. acuminatum)亲缘关系最近,可信度达99%;最后通过根部接种试验,接种苜蓿根部发病症状与田间根腐病发病症状一致,鉴定结果显示本试验研究的引起紫花苜蓿根腐病的病原菌为镰刀菌属锐顶镰刀菌(F. acuminatum)。     

郑州地区3种冷季型草坪草根部入侵真菌鉴定

研究不同草坪草根部入侵真菌的种类对草坪病害鉴定与防治具有重要意义。本试验以河南省郑州市内8个公共草坪上的3种冷季型草坪草高羊茅(Festuca arundinacea),草地早熟禾(Poa pratensis)和多年生黑麦草(Lolium perenne)为研究材料,于2013年春季,在8个混播草坪上采用"Z"字形法随机进行根部取样,每种草在各采样地内分别取12株根样,并在实验室内分离鉴定其根部入侵真菌种类。结果表明:从288株根样中共分离到13种真菌,其中6种真菌为3种草根部共同的入侵真菌,包括:细交链孢(Alternaria alternata)、尖孢镰孢(Fusarium oxysporum)、柱孢(Cylindrocarpon destructuns)、枝孢(Cladosporium herbarum)、多隔镰孢(F.decemcellulare)、德氏霉(Drechslera sp.)。郑州地区冷季型草坪草根部入侵的主要病原真菌是细交链孢、尖孢镰孢和枝孢,不同地点和草种根部主要的入侵真菌种类有明显的不同,根部带菌率最高的是多年生黑麦草。     

变灰青霉BIBA-G563对紫花苜蓿镰刀菌根腐病的防效

从黑龙江省安达市黑龙江八一农垦大学草业科学系试验田健康紫花苜蓿根部分离出1株拮抗真菌BIBA-G563,对其进行形态学观察和分子生物学鉴定,并研究其对紫花苜蓿根腐病病原真菌镰刀菌的抑制能力及盆栽防治根腐病的效果。经鉴定,拮抗真菌BIBA-G563为变灰青霉。BIBA-G563与腐皮镰刀菌、尖孢镰刀菌、层出镰刀菌、轮枝镰刀菌四种病原真菌的第9 d平板对峙抑制率在28.62%～64.67%之间,无菌发酵液对四种病原菌菌丝生长的抑制率在10.34%～15.98%之间,对四种病原菌孢子萌发的抑制率在16.95%～35.95%之间,对盆栽紫花苜蓿镰刀菌根腐病防效为75.40%。表明生防真菌BIBA-G563能够有效抑制紫花苜蓿根腐病病原菌,具有良好的开发前景。     

1株田无链霉菌T2的鉴定及其对紫花苜蓿根腐病的防效研究

为了验证从紫花苜蓿根际土壤中分离出的1株对根腐病致病菌——镰刀菌(Fusarium spp.)具有抑制作用的菌株T2,试验通过形态学、生理生化试验、16S rDNA基因鉴定确定菌株种属，通过平板对峙试验及孢子萌发试验验证菌株T2对镰刀菌的抑菌能力，通过盆栽试验验证菌株T2对紫花苜蓿根腐病的防效。结果表明：经形态学观察、生理生化鉴定及16S rDNA基因序列分析，确定菌株T2为田无链霉菌(Streptomyces tanashiensis)。在平板对峙试验中田无链霉菌T2对腐皮镰刀菌(Fusarium solani,F.solani)、尖孢镰刀菌(Fusarium oxysporum,F.oxysporum)、轮枝镰刀菌(Fusarium verticillioides,F.verticillioides)的抑制率分别为37.33%、53.33%和37.00%。田无链霉菌T2无菌发酵液对腐皮镰刀菌、尖孢镰刀菌及轮枝镰刀菌孢子萌发抑制率分别为42.11%、43.94%和44.96%。盆栽试验中，田无链霉菌T2对苗期紫花苜蓿根腐病的防效为50.21%。说明单独接种田无链霉菌T2可以促进紫花苜...     

郑州地区冷季型草坪草根腐病病原鉴定

通过对郑州地区冷季型草坪草根腐病病原物分离鉴定发现,主要病原物有镰刀菌(Fusarium spp.)、链格孢菌(Alternaria spp.)、丝核菌(Rhizoctonia spp.)、德氏霉菌(Drechslera spp.)、弯孢菌(Curvularia spp.)和腐霉菌(Pythium spp.)6大类。其中,茄腐镰刀菌(Fusarium solani)和立枯丝核菌(Rhizoctonia solani)(AG-4)的分离频率最高,致病力比较强,为优势病原。     

两株紫花苜蓿根际芽孢杆菌的筛选及生防效果研究

从紫花苜蓿根际土中分离出2株芽孢杆菌(Bacillus spp.)菌株CYY-6和CYY-42,并评价了其对尖孢镰刀菌(Fusarium oxysporum)的抑制作用,及其对由尖孢镰刀菌引起的苜蓿根腐病的防治效果。经过形态观察、16S rDNA序列分析和生理生化鉴定,确定菌株CYY-6为地衣芽孢杆菌(Bacillus licheniformis),CYY-42为短短芽孢杆菌(Brevibacillus brevis)。同时检测发现这2株芽孢杆菌均能不同程度地合成嗜铁素。在平板对峙实验中,2株菌株均能对尖孢镰刀菌产生拮抗作用,并产生抑菌带,抑菌率分别为49.3%和56.3%。且其发酵液可以致使尖孢镰刀菌菌丝及孢子生长异常,出现菌丝扭曲、断裂、破碎,孢子数量明显减少等现象。在盆栽实验中,2株菌株对由尖孢镰刀菌引起的紫花苜蓿根腐病的相对防效分别为57.55%和64.03%,在紫花苜蓿根腐病防治方面具有较好的应用潜力。     

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.     

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.     

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.     

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