白粉菌侵染Pm21近等基因系的早期蛋白组差异研究   总被引：2，自引：0，他引：2  

李达  李强  关西贞  张卫东  田纪春 《华北农学报》2012,(1):5-11

分析接种小麦白粉菌E09后Pm21近等基因系及其亲本叶片的总蛋白质差异,从蛋白质组调控角度揭示Pm21基因抗白粉病的抗病机理。以Pm21近等基因系及其亲本百农3217为试材,显微观察接种后5 d内的叶片表面细胞与白粉菌孢子表型差异;采用非离子去污剂提取总蛋白质,运用双向电泳、质谱分析与数据库检索技术,比较Pm21近等基因系及其亲本接菌第4天后的叶片总蛋白质差异。超显微和显微观察结果表明,12 h后百农3217叶细胞表面与Pm21近等基因系相比褶皱明显;在接菌第2天后,在Pm21近等基因系白粉菌孢子生长明显慢于其亲本表面孢子,且孢子侵入个数较少,不萌发现象较多;在第4天后Pm21近等基因系被侵染叶片细胞出现过敏性反应;通过MALDI-TOF-MS/MS质谱测序和MASCOT软件序列分析,注释了15个表达上调蛋白点;功能分类表明,差异蛋白质分别参与叶绿体光合呼吸作用(40%)、放氧作用(13%)、代谢物合成(20%)、抗病、动力蛋白、运输等生理功能。蛋白质组水平发现,光合呼吸蛋白和叶谷氨酰胺合成酶在Pm21近等基因系感病3～4 h后就大量表达,可能是导致叶片明显变绿原因;叶谷氨酰胺合成酶的增加与光合呼吸蛋白的增加密不可分;光合呼吸蛋白和抗坏血酸过氧化物酶的上调表达为植物体在高H2O2浓度下正常生长提供了保障。另外,β-1,3-葡聚糖苷酶的过量表达也是Pm21基因前期抵御白粉菌侵染有效物质。钾离子通道蛋白可能在抗病防御过程中起到了信号转导的作用。  相似文献   

木材顺纹压缩率与PDR的影响机制研究     

张燕  宋魁彦*  佟达 《林业机械与木工设备》2012,(11):35-37

木材顺纹压缩弹塑性和PDR(永久变形率)变化规律及机理研究是攻克木材顺纹压缩与多维弯曲技术的核心.从木材的树种、密度、软化处理条件、化学组分和相对结晶度以及压缩速度几个方面,分析了木材顺纹压缩过程中这些因素对木材顺纹压缩率和PDR的影响机制.  相似文献   

Qualitative and quantitative polymerase chain reaction (PCR) for detection of Leishmania in spleen samples from naturally infected dogs     

Solcà Mda S  Guedes CE  Nascimento EG  Oliveira GG  dos Santos WL  Fraga DB  Veras PS 《Veterinary parasitology》2012,184(2-4):133-140

Because infected dogs are widely considered to be the main domestic reservoir for Leishmania infantum (syn Leishmania chagasi) parasites in Brazil, the diagnosis of canine visceral leishmaniasis (CVL) must be made both accurately and promptly. The present study attempted to standardize a conventional polymerase chain reaction (cPCR) protocol for the detection of L. infantum DNA in canine spleen samples. Quantitative PCR (qPCR) technique was used to confirm the presence of Leishmania DNA in the canine spleen fragments. A comparison was made between the efficacies of these molecular diagnostic techniques and conventional parasitological and serological methods. cPCR protocols for spleen samples were standardized using primers that amplify a 145 bp fragment, located at the parasite kinetoplast minicircle. The genus specificity of the cPCR protocol was assessed by its inability to amplify the DNA of other common canine pathogens, such as Ehrlichia canis, Babesia canis, Toxoplasma gondii and Trypanosoma cruzi. cPCR protocol sensitivity was tested by assessing the reaction detection limit, determined to be 10 fg of L. infantum reference strain DNA, which corresponds to a range of 0.03-0.1 parasites per fragment. Standardized cPCR protocol was used to detect the presence of Leishmania in 45 dog spleen samples. Our results showed that 40% of the spleen fragment cultures were positive for Leishmania parasites, 58% of the dog serum samples tested positive using ELISA, and parasite DNA was detected in 44% using qPCR, while 47% of the spleen samples using cPCR. Diagnostic methods performance was assessed and revealed a better degree of ascertainment for cPCR when compared to other diagnostic methods. The sensitivity of ELISA was 83.3%, qPCR was 83.3%, and cPCR was 88.9%; PPV for ELISA was 57.7%, qPCR was 75% and cPCR was 76.2%; the Kappa coefficients were found to be 0.40 (fair) for ELISA, 0.64 (substantial) for qPCR and 0.68 (substantial) for cPCR. In both oligosymptomatic and polysymptomatic dogs, cPCR revealed the better performance analysis when compared to other diagnostic methods. The findings presented herein establish cPCR as the most indicated test to detect Leishmania when compared to the other two diagnostic methods evaluated. Despite the fact that the qPCR protocol provides a highly accurate quantification of parasites when targeting the SSU rRNA gene, this technique does not significantly improve the diagnosis of CVL when compared with the performance of the cPCR protocol, which focused on the kinetoplast minicircle.  相似文献   

Increased incidence of DNA amplification in follicular than in uterine and blood samples indicates possible tropism of Neospora caninum to the ovarian follicle     

Silva AF  Rangel L  Ortiz CG  Morales E  Zanella EL  Castillo-Velázquez U  Gutierrez CG 《Veterinary parasitology》2012,188(1-2):175-178

This study evaluated the presence of Neospora caninum in ovarian follicle aspirates and uterine flushes obtained from N. caninum seropositive dairy cows. Ninety-two cows that aborted within the previous 90 days were sampled to determine the presence of antibodies against N. caninum. Thirteen seropositive cows were chosen for collection of blood leukocytes, uterine flushes (UF; n=12) and follicular aspirates (OPU; n=13). Samples were centrifuged and the cellular sediment from the follicular fluid, uterine flushes and blood leukocytes were used for DNA extraction and PCR. Follicular aspirates had the highest frequency of DNA amplification for N. caninum (p<0.05, 92.3%; 12/13). Whereas uterine (4/12) and blood leukocyte (5/13) samples had similar (p>0.05) rate of positive results. Nonetheless, there was no agreement between blood leukocytes and follicular samples taken from the same animal (Cohen Kappa=-0.16). Similarly, blood leukocytes and uterine results had moderate agreement between them (Cohen Kappa=0.47). This study indicates that N. caninum is present in the ovarian follicle and uterus of seropositive cows, suggesting a possible risk of neosporosis transmission between females during oocyte and embryo collection and transfer. Hence, precautions should be taken to minimize the risk of N. caninum transmission. Furthermore, the high incidence of positive results in follicle samples, that exceeded that of their paired blood leukocytes, suggests a possible tropism of N. caninum for the ovarian follicle.  相似文献   

Partial characterization of an atypical family I inorganic pyrophosphatase from cattle tick Rhipicephalus (Boophilus) microplus     

Costa EP  Campos E  de Andrade CP  Façanha AR  Saramago L  Masuda A  Vaz Ida S  Fernandez JH  Moraes J  Logullo C 《Veterinary parasitology》2012,184(2-4):238-247

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Saccharomyces boulardii reduces infection intensity of mice with toxocariasis     

de Avila LF  Conceição FR  Telmo Pde L  Dutra GF  de los Santos DG  Martins LH  Berne ME  da Silva PE  Scaini CJ 《Veterinary parasitology》2012,187(1-2):337-340

Several studies have shown the benefit of the use of probiotics in the prevention and treatment of diseases; however, few of them have investigated the effect of probiotics on parasitosis. In this study, the effect of Saccharomyces boulardii on the intensity of infection of mice with toxocariasis was evaluated. The animals were fed with a diet supplemented with S. boulardii for 15 days before inoculation with Toxocara canis eggs and for 2 or 60 days post-inoculation. S. boulardii promoted a reduction of approximately 36% in the average number of recovered T. canis larvae, suggesting that it can be used as an alternative to help control toxocariasis.  相似文献   

A new type of pterocarpanquinone that affects Toxoplasma gondii tachyzoites in vitro     

Portes Jde A  Netto CD  da Silva AJ  Costa PR  DaMatta RA  dos Santos TA  De Souza W  Seabra SH 《Veterinary parasitology》2012,186(3-4):261-269

Toxoplasma gondii, the agent of Toxoplasmosis, is an obligate intracellular protozoan able to infect a wide range of vertebrate cells, including nonprofessional and professional phagocytes. Therefore, drugs must have intracellular activities in order to control this parasite. The most common therapy for Toxoplasmosis is the combination of sulfadiazine and pyrimethamine. This treatment is associated with adverse reactions, thus, the development of new drugs is necessary. In previous studies, naphthoquinone derivatives showed anti-cancer activity functioning as agents capable of acting on groups of DNA, preventing cancer cells duplication. These derivatives also display anti-parasitic activity against Plasmodium falciparum and Leishmania amazonensis. The derivative pterocarpanquinone tested in this work resulted from the molecular hybridization between pterocarpans and naphtoquinone that presents anti-tumoral and anti-parasitic activities of lapachol. The aim of this work was to determine if this derivative is able to change T. gondii growth within LLC-MK2 cells. The drug did not arrest host cell growth, but was able to decrease the infection index of T. gondii with an IC(50) of 2.5 μM. Scanning and transmission electron microscopy analysis showed morphological changes of parasites including membrane damage. The parasite that survived tended to encyst as seen by Dolichos biflorus lectin staining and Bag-1 expression. These results suggest that pterocarpanquinones are drugs potentially important for the killing and encystment of T. gondii.  相似文献   

Enrofloxacin is able to control Toxoplasma gondii infection in both in vitro and in vivo experimental models     

Barbosa BF  Gomes AO  Ferro EA  Napolitano DR  Mineo JR  Silva NM 《Veterinary parasitology》2012,187(1-2):44-52

Currently, toxoplasmosis is treated with sulfadiazine and pyrimethamine. However, this treatment presents several adverse side effects; thus, there is a critical need for the development and evaluation of new drugs, which do not present the same problems of the standard therapy. Enrofloxacin is a fluoroquinolone antibiotic known to control infection against several bacteria in veterinary medicine. Recently, this drug has demonstrated protective effects against protozoan parasites such as Neospora caninum. The present study aimed to determine the effect of enrofloxacin in the control of Toxoplasma gondii infection. For this purpose, human foreskin fibroblast (HFF) cells were infected with T. gondii RH strain and treated with sulfadiazine, penicillin/streptomycin, pyrimethamine, or enrofloxacin. Following treatment, we analyzed the infection index, parasite intracellular proliferation and the number of plaques. Additionally, tissue parasitism and histological changes were investigated in the brain of Calomys callosus that were infected with T. gondii (ME49 strain) and treated with either sulfadiazine or enrofloxacin. Enrofloxacin was able to reduce the infection index, intracellular proliferation and the number of plaques in HFF cells infected by T. gondii in comparison with untreated or penicillin/streptomycin-treated ones. Enrofloxacin was more protective against T. gondii in HFF infected cells than sulfadiazine treatment (P<0.001). In addition, pyrimethamine, enrofloxacin or the associations of sulfadiazine plus pyrimethamine, enrofloxacin plus sulfadiazine or enrofloxacin plus pyrimethamine-treatments were able to reduce the plaque numbers in HFF cells infected by T. gondii when compared to medium, penicillin/streptomycin or sulfadiazine alone. In vivo experiments demonstrated that enrofloxacin diminished significantly the tissue parasitism as well as the inflammatory alterations in the brain of C. callosus infected with T. gondii when compared with untreated animals. Based on our findings, it can be concluded that enrofloxacin is a potential alternative drug for the treatment of toxoplasmosis.  相似文献   

Effect of vascularity of corpus luteum in progesterone production of mares     

Renata Cristina Uliani  Mateus José Sudano  Eunice Oba  Luciano Andrade Silva  Marco Antonio Alvarenga 《Journal of Equine Veterinary Science》2012

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Effects of lactoferrin supplementation to milk- and caseinate-based extenders on sperm motility,membrane integrity and nitric oxide levels of cooled stallion semen     

H.S. Martins  M.R. Souza  C.F.A.M. Penna  G.C. da Silva  S.F. Côrtes  R. Stahlberg  M.F. Brito  M.A. Lagares 《Journal of Equine Veterinary Science》2012

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