The expression of antigens in transgenic plants has increasingly been used as an alternative to the classical methodologies for the development of experimental vaccines.This paper reports here the development of a nov...The expression of antigens in transgenic plants has increasingly been used as an alternative to the classical methodologies for the development of experimental vaccines.This paper reports here the development of a novel oral immunization system for foot-and-mouth disease (FMD) in transgenic maize with two serotypes of the structural protein VP1 of the foot-and-mouth disease virus (FMDV) viz.,O-and Asia 1-type,respectively.The transgenic plantlets were identified and investigated by polymerase chain reaction (PCR),Southern blot,and real-time PCR.Moreover,it was found that the VP1 genes in transgenic plants could be transmitted stably to the next generation through PCR detection.To our knowledge,this is the first report in an attempt to induce a protective systemic antibody response in animals by feeding the transgenic plants in which two serotypes antigen protein of FMDV expressed together.Results of the experiment provide the possibility of using plant-based vaccines as feedstuff or feedstuff additives.展开更多
[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] Th...[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] The optimal fermentation conditions were determined by single factor test and orthogonal design. [ Result J The optimum fermentation conditions are as following: inoculum weight ratio (Aspergillus oryzae vs Trichoderma vinde), 1:1 ; inoculum size, 30% ; water content, 40% ; fermentation time, 96 h; and fermentation temperature, 30℃. Under these conditions, glucosinolates were reduced by 90.71% and neutral detergent fibers were degraded by 20.65%. [ Condusion] In laboratory, solid state fermentation with Aspergillus oryzae and Trichoderrna viride can be used to produce high-quality rapeseed meal.展开更多
In recent years, subgroup J avian leukosis virus (ALV-J) has been found to frequently infect layers in China. This virus is responsible for economic losses due to both mortality and decreased performance in chickens...In recent years, subgroup J avian leukosis virus (ALV-J) has been found to frequently infect layers in China. This virus is responsible for economic losses due to both mortality and decreased performance in chickens. In this study, 45-d-old cloned flee-range layers were suspected to be infected with ALV and other immunosuppressive diseases because their feathers were unkempt and their growth rate was impaired. To estimate the infection status and determine the source of ALV-J in the flock, 30 cloacal swabs were randomly collected to measure the p27 antigen level by enzyme-linked immunosorbent assay (ELISA). Among the birds that were tested, 87% (26/30) were positive. In addition, 6 anticoagulant blood samples were aseptically collected at random from the flock when the layers were 60 d old. These samples were centrifuged to obtain the leukocytes, which were then used to inoculate chicken embryo fibroblast (CEF) cells for the identification of ALV-J by indirect immunofluorescence (IFA). Of the samples tested, 100% (6/6) were positive. The flock's production performance was also investigated, and 10 layers were necropsied to evaluate pathological changes at 115 d of age. The flock never laid eggs even though they reached the age of the first laying (110 d). Furthermore, there were pathological changes present, including atrophy of the thymus and bursa of Fabricius, undeveloped ovaries, glandular stomach haemorrhage, and hepatosplenomegaly. Paraffin-embedded sections of intumescent liver and spleen were prepared for antigen localisation using IFA. Positive signals were prevalent in paraffin-embedded sections of the intumescent liver and spleen. Furthermore, provirus DNA was extracted from 4 cloned flee-range layers, and 2 patemal parents (HR native cocks), and the gp85 gene of ALV-J was amplified by PCR to analyse the genetic variation. The results of the autogenous variation analysis showed that the 6 strains were 98.5-99.7% homologous. This study indicated that there was persistent infection with ALV-J by dynamic inspection, which seriously reduced the production performance of the flock. In addition, the genetic variation analysis showed that ALV-J in the flock was more likely to have originated from the paternal parent, the HR native cock.展开更多
基金supported by the National Natural Science Foundation of China (30800687 and 31071434)the Applied Basic Research Program of Sichuan Provincial Science and Technology Department,China (2008JY0096)+1 种基金the Foundation for Young Scientists of Sichuan Provincial Education Department,China(09ZB051)the Youth Innovation Project of Sichuan Agricultural University,China,the Postdoctoral Project of Sichuan Agricultural University,China
文摘The expression of antigens in transgenic plants has increasingly been used as an alternative to the classical methodologies for the development of experimental vaccines.This paper reports here the development of a novel oral immunization system for foot-and-mouth disease (FMD) in transgenic maize with two serotypes of the structural protein VP1 of the foot-and-mouth disease virus (FMDV) viz.,O-and Asia 1-type,respectively.The transgenic plantlets were identified and investigated by polymerase chain reaction (PCR),Southern blot,and real-time PCR.Moreover,it was found that the VP1 genes in transgenic plants could be transmitted stably to the next generation through PCR detection.To our knowledge,this is the first report in an attempt to induce a protective systemic antibody response in animals by feeding the transgenic plants in which two serotypes antigen protein of FMDV expressed together.Results of the experiment provide the possibility of using plant-based vaccines as feedstuff or feedstuff additives.
基金funded by the grants from National High Technology Research and Development Program of China (2009A-A02Z210)Key Technology R&D Program of Qingdao City,China (07-2-1-25-nsh)
文摘[Objective] To optimize solid state fermentation conditions of rapeseed meal and thus to reduce glucosinolates and neutral detergent fibers by mixed cultures of Aspergillus oryzae and Tnchoderrna viride. [ Method ] The optimal fermentation conditions were determined by single factor test and orthogonal design. [ Result J The optimum fermentation conditions are as following: inoculum weight ratio (Aspergillus oryzae vs Trichoderma vinde), 1:1 ; inoculum size, 30% ; water content, 40% ; fermentation time, 96 h; and fermentation temperature, 30℃. Under these conditions, glucosinolates were reduced by 90.71% and neutral detergent fibers were degraded by 20.65%. [ Condusion] In laboratory, solid state fermentation with Aspergillus oryzae and Trichoderrna viride can be used to produce high-quality rapeseed meal.
文摘In recent years, subgroup J avian leukosis virus (ALV-J) has been found to frequently infect layers in China. This virus is responsible for economic losses due to both mortality and decreased performance in chickens. In this study, 45-d-old cloned flee-range layers were suspected to be infected with ALV and other immunosuppressive diseases because their feathers were unkempt and their growth rate was impaired. To estimate the infection status and determine the source of ALV-J in the flock, 30 cloacal swabs were randomly collected to measure the p27 antigen level by enzyme-linked immunosorbent assay (ELISA). Among the birds that were tested, 87% (26/30) were positive. In addition, 6 anticoagulant blood samples were aseptically collected at random from the flock when the layers were 60 d old. These samples were centrifuged to obtain the leukocytes, which were then used to inoculate chicken embryo fibroblast (CEF) cells for the identification of ALV-J by indirect immunofluorescence (IFA). Of the samples tested, 100% (6/6) were positive. The flock's production performance was also investigated, and 10 layers were necropsied to evaluate pathological changes at 115 d of age. The flock never laid eggs even though they reached the age of the first laying (110 d). Furthermore, there were pathological changes present, including atrophy of the thymus and bursa of Fabricius, undeveloped ovaries, glandular stomach haemorrhage, and hepatosplenomegaly. Paraffin-embedded sections of intumescent liver and spleen were prepared for antigen localisation using IFA. Positive signals were prevalent in paraffin-embedded sections of the intumescent liver and spleen. Furthermore, provirus DNA was extracted from 4 cloned flee-range layers, and 2 patemal parents (HR native cocks), and the gp85 gene of ALV-J was amplified by PCR to analyse the genetic variation. The results of the autogenous variation analysis showed that the 6 strains were 98.5-99.7% homologous. This study indicated that there was persistent infection with ALV-J by dynamic inspection, which seriously reduced the production performance of the flock. In addition, the genetic variation analysis showed that ALV-J in the flock was more likely to have originated from the paternal parent, the HR native cock.