苜蓿假盘菌与苜蓿叶片亲和性互作的超微结构特征

从细胞角度揭示苜蓿假盘菌与苜蓿叶片亲和性互作机制。以两者亲和性互作系统为研究材料,利用透射电子显微镜技术研究了苜蓿假盘菌与苜蓿叶片亲和性互作的超微结构特征。结果表明,病菌侵入寄主组织后,菌丝直接穿透寄主细胞壁进入寄主细胞形成胞内菌丝,以胞内生长并向相邻细胞扩展。病菌菌丝穿透寄主细胞壁时,菌丝中的液泡给予了较大的压力帮助其穿透。进入寄主细胞内的病菌菌丝,被内陷的寄主原生质膜包裹,菌丝与质膜始终是隔离的,寄主原生质膜和细胞壁之间沉积电子致密度深的物质。病菌菌丝不断地在寄主原生质膜区域扩展,伴随菌丝体在寄主细胞内的不断扩大,周围的原生质膜也相应扩大其面积,但始终将寄主原生质与菌丝体隔开,而脱离质膜的菌丝形成菌丝鞘包裹。随侵染程度的增加,未被穿透的寄主原生质膜区域逐步被降解。病菌侵染叶绿体等细胞器时,首先是菌丝鞘与叶绿体等细胞器膜相连,然后降解其基粒片层结构,被降解的细胞器组织沿菌丝和胞壁周围沉积。侵染后期,菌丝胞内和胞外扩展,但处在细胞降解物中的菌丝显示较厚的细胞壁,寄主细胞内充满了大量的黑色物质和结晶状的颗粒物。     

In vivo and ex vivo assessment of the interaction between ivermectin and danofloxacin in sheep

The impact of an efflux pump-related interaction between ivermectin and danofloxacin on their intestinal transport (ex vivo) and disposition kinetics (in vivo) was assessed. Eighteen male Corriedale sheep were randomly assigned to one of three groups. Animals in Group A received 0.2mg/kg ivermectin by SC injection, those in Group B were given 6 mg/kg danofloxacin SC on two occasions 48 h apart and those in Group C were treated with both compounds at the same rates. Plasma concentrations of ivermectin and danofloxacin were measured by HPLC using fluorescence detection. Ex vivo intestinal drug transport activity was measured by the use of the Ussing chamber technique. Plasma concentrations of ivermectin in the first 6 days after injection tended to be higher in Group C than Group A. Contemporaneous treatment with ivermectin significantly increased systemic exposure to danofloxacin (AUC values were 32-35% higher) and prolonged the elimination half-life of danofloxacin (40-52% longer). Ex vivo, incubation with ivermectin significantly decreased the efflux transport of rhodamine 123, a P-glycoprotein substrate, in sheep intestine, but no significant effect of danofloxacin on transport activity was observed. Evaluation of the interaction of danofloxacin with the breast cancer resistance protein (BCRP) showed that pantoprazole and ivermectin significantly decreased danofloxacin secretion in the rat intestine. Thus, the ivermectin-induced reduction of danofloxacin efflux transport observed in this study may involve BCRP activity but the involvement of P-glycoprotein cannot be ruled out.     

Prevalence of Salmonella and Yersinia in free-living pied flycatchers (Ficedula hypoleuca) in central Spain

Salmonella and Yersinia are important enteropathogens in poultry and can affect birds of all ages, including embryos. These food-borne zoonotic enteropathogens are of great economic and medical concern worldwide and are intensely studied in poultry. Information regarding the prevalence of these bacteria in wild birds is scarce and biased toward avian species ecologically linked to humans, which have often been incriminated as both reservoirs and disseminators of these enteropathogens. The prevalence of Salmonella and Yersinia recovered from both the feces and eggs in a population of female pied flycatchers (Ficedula hypoleuca) breeding in nest-boxes in central Spain was evaluated. Salmonella arizonae was recovered from the feces of one female but was not recovered from eggs. Yersinia was not detected in either the feces or eggs. These results may suggest that Salmonella and Yersinia may be uncommon in this population studied and may indicate that these birds are unlikely reservoirs of Salmonella and Yersinia.     

一株无血凝性兔出血症病毒的分离鉴定及其衣壳蛋白VP60基因的克隆和序列分析

从临床病料中分离鉴定出1株无血凝性的致病性兔出血症病毒（RHDV），命名为Yaan-1。HA试验测得该毒株原代及经兔体传3代后的肝毒在25℃、37℃和4℃时均无血凝性；但在琼脂扩散试验和对流免疫电泳中均可见分离株与常规RHDV高免血清形成清晰的沉淀线；电镜观察可见30nm左右的病毒粒子。采用RT-PCR方法将Yaan-1株的衣壳蛋白VP60基因进行了克隆测序，序列分析表明VP60基因序列全长1740bp，编码579个氨基酸，测序结果提交GenBank（注册号为DQ069280），并与世界各国的常规RHDV分离株进行比较，结果表明核苷酸同源性为90．0％～98．0％，氨基酸同源性为94．1％～99．0％，氨基酸变异多发生在衣壳蛋白C、E区；同时对Yaan-1株的分子量、等电点、疏水性、跨膜区等分子结构特征进行了分析。本研究首次报道了无血凝性RHDV中国分离株，丰富了地方毒株资源，为明确我国地方株的分子流行病学，以及对VP60蛋白分子特征研究提供了新材料。     

浅谈二噁英的检验技术

1　检测二口恶英的技术难度食品和饲料中二口恶英及其类似物的检验属于超痕量级检验,通常以ｎｇ/ｋｇ和ｐｇ/ｋｇ表示。它比其他污染物含量低千倍到亿倍以上,检测难度很大,其多组分即同系物、异构体的化学前处理技术几乎包罗近代最先进的手段,涉及分离、富集、提取、净化、清洗和浓缩装置,还包含自动凝胶色谱(ＡｕｔｏｍａｔｅｄＧｅｌＰｅｒｍｅａｔｉｏｎＣｈｒｏｍａｔｏｇｒａｐｈｙ)、反相ＨＰＬＣ、玻璃色谱柱等。检测过程需要尖端大型分析仪器的支持,定量分析依靠ＨＲＧＣ/ＨＲＭＳ,对仪器的灵敏度、选择性和特异性要求极高,是现今分…     

鸡球虫耐药性的机理检测方法及其延缓措施

文章对鸡球虫耐药性的产生机制、检测方法及其延缓措施的研究进展进行了概述。     

Use of comparative proteomics to identify the effects of creatine pyruvate on lipid and protein metabolism in broiler chickens

Four hundred male chickens were selected to study the effects of pyruvate (Pyr), creatine pyruvate (CrPyr) and creatine (Cr) on the expression of hepatic mitochondrial and cytoplasm proteins associated with lipid and protein metabolism. Mitochondrial purification was accomplished using the two-step differential centrifugation and density gradient method, and the activities of organelle-specific marker enzymes were determined to assess the purity of the mitochondria. Proteins were extracted and fractionated by two-dimensional electrophoresis and the differential protein spots were assessed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. CrPyr reduced fatty acid accumulation by down-regulating adipose differentiation-related protein, inhibited ATP synthase expression, and reduced cholesteryl ester transfer protein (CETP) expression, thus reducing the levels of high density lipoprotein and triglycerol (TG) levels (thereby lowering fat and cholesterol deposition). CrPyr increased the expression of eukaryotic translation initiation factor (eIF) 2B, calreticulin (CRT) and eIF3a, thus promoting protein synthesis. CrPyr up-regulated the expression of fatty acid-binding proteins, CETP and apolipoprotein A-IV in cytoplasmic extracts, and these proteins accelerated the decomposition of fatty acids and TG, thus reducing fat deposition. In conclusion, CrPyr plays an important role in lipolysis and protein synthesis, and this effect was more pronounced than was the effect of Pyr and Cr.     

Septicaemia secondary to infection by Corynebacterium macginleyi in an Indian python (Python molurus)

A seven-year-old female Indian python (Python molurus) weighing about 35kg was euthanased after several clinical episodes of stomatitis, pneumonia, ophthalmitis and dystocia over a period of four years. The animal had been maintained in a terrarium in a circus truck at an adequate temperature. During shows, however, the snake was considered to be exposed to stressful conditions for several hours at a time at low temperatures and with noise and bright lights. A post-mortem examination indicated ulcerative stomatitis, osteomyelitis, severe pneumonia and numerous granulomata and multifocal necrosis in stomach and spleen. Corynebacterium macginleyi was isolated in pure culture from the ulcerative stomatitis, and mixed with Stenotrophomonas maltophilia from the lungs and spleen. The findings indicated that the snake had died from a septicaemic process caused by C. macginleyi, probably originating from the stomatitis. The role of S. maltophilia as a secondary agent is discussed. The stress of the circus show and poor husbandry may have predisposed the animal to infection and septicaemia. This is the first report of C. macginleyi causing disease in a snake.     

In vitro antiviral activity of dehydroepiandrosterone and its synthetic derivatives against vesicular stomatitis virus

In this work the antiviral activity of 20 dehydroepiandrosterone (DHEA) analogs with different substituents at positions C-3, C-15, C-16 and C-17 were evaluated against vesicular stomatitis virus (VSV) in Vero cell cultures. The selectivity indexes (SI) obtained with DHEA and epiandrosterone (EA) were 50 and 72.6, respectively. The work showed that the compounds 21-norpregna-5,17(20)-dien-3β,16α-diyl-diacetate, 17,17-ethylendioxyandrostan-5,15-dien-3β-ol and 3β-hydroxypregn-17(20)-en-16-one had higher SI values than ribavirin, which was used as a reference drug. The antiviral mode of action of DHEA was also investigated against VSV replication in Vero cells, and time of addition experiments showed that DHEA mainly affected a late event in the virus growth cycle. Analysis of RNA and protein synthesis indicated that DHEA adversely affected positive strand RNA synthesis and viral mature particle formation.