Mycoplasma gallisepticum HS strain(MG-HS)is a pathogen that causes chronic respiratory disease(CRD)in chicken,which is characterized by host respiratory inflammatory damage,brings huge economic losses to the poultry i...Mycoplasma gallisepticum HS strain(MG-HS)is a pathogen that causes chronic respiratory disease(CRD)in chicken,which is characterized by host respiratory inflammatory damage,brings huge economic losses to the poultry industry.Recently,emerging Chinese herbal medicines(CHM)have been used to treat CRD.This study was aimed to investigate the preventive and therapeutic effects and their potential mechanisms of Chinese herbal medicinal formulae(CHMF),which consisted of 10 kinds of Chinese herbal medicine including Scutellaria,Houttuynia cordate and licorice,on MGinduced CRD in chickens.With respect to the preventive effect,the results showed that CHMF could effectively recover the MG-induced decrease on body weight and feed conversion ratio.Histopathological analysis showed that both prevention and treatment of CHMF could significantly alleviate the severe respiratory inflammation induced by MG infection.Moreover,compared with the MG infection group,both the prevention groups and the treatment groups of CHMF could effectively reduce the expression of MG adhesion protein(p MGA1.2)to inhibit the proliferation of MG,and thus effectively inhibit the expression of MG-induced inflammatory factors interleukin-1β(IL-1β)and tumour necrosis factor-α(TNF-α).In summary,these findings confirm that CHMF can protect chickens from various tissue damage caused by MG infection and has no adverse effects on the performance of chickens in the short term.And its efficacy against MG is equal to or better than that of tiamulin.展开更多
The study and entire laboratory works were conducted from December 2014 to April 2015 in National Veterinary Institute, Bishoftu, Ethiopia. Formaldehyde inactivated Montanide ISA70 based Mycoplasma gallisepticum (MG) ...The study and entire laboratory works were conducted from December 2014 to April 2015 in National Veterinary Institute, Bishoftu, Ethiopia. Formaldehyde inactivated Montanide ISA70 based Mycoplasma gallisepticum (MG) trial vaccine strain was confirmed the identity with known primer using PCR from locally isolates of National Veterinary Institute of Ethiopia. This study was aimed to develop formaldehyde inactivated Montanide ISA70 based MG vaccine in Ethiopia. It can help to device strategies in controlling the disease mainly through developing more effective vaccine which will replace the currently being imported vaccines by some farms. After culturing procedure, oil based inactivated MG trial vaccine was produced in suitable clean and secure accommodation. In this study, among different isolates, local isolate of Samuel farm in NVI was prepared and evaluated in chickens. The amount of immune antigen per 0.5 ml of the dose was 107 Colony forming units (CFU) of the bacteria. The trail vaccine was prepared and evaluated at the age of 16 weeks of chickens;the chickens were randomly divided into three groups (A, B and C), each having twenty birds (10 male and 10 female). Each of group B was vaccinated group of imported-live vaccine with 30 μl intraocularly for comparing with inactivated trial vaccine, each bird of group C was inoculated with 0.5 ml indigenous or trial vaccine subcutaneously at mid neck region and group A was used as a control then challenge tests were performed. After challenge test, among non-vaccinated chickens (control or group A) 2 chickens were died (10%), thicken and cloudy appearance of the air sac showed 18 (90%), 2 chickens were not showed thickened and cloudy air sack (10%). Although among vaccinated group (inactivated vaccine or group C), all chickens did not show clinical signs or post mortem changes (100%). From attenuated imported live vaccine (group B), no clinical signs or post mortem changes were observed (100%). It was concluded that oil based MG vaccine induces protective level of anti MG antibodies in chickens.展开更多
The detailed genetic analysis of mycoplasmas has long been hampered by the lack of appropriate tools for genetic manipulation. In this study, the transposon vector, mini-Tn4001tetM, was constructed containing the tnp ...The detailed genetic analysis of mycoplasmas has long been hampered by the lack of appropriate tools for genetic manipulation. In this study, the transposon vector, mini-Tn4001tetM, was constructed containing the tnp gene, encoding a transposase gene in Staphylococcus aureus, two copies of the IS256 inverted repeat sequence (inner and outer) and the tetM gene, from the Enterococcus faecalis Tn916 transposon, conferring resistance to tetracycline. This vector was electro-transformed into Mycoplasma gallisepticum (MG). The recombinant cells were screened by tetracycline selection. The results indicated that the transposon vector could replicate in MG strain R by successive passages, indicating that MG is a potential vector for expressing protective antigens of other pathogens.展开更多
【目的】建立基于鸡毒支原体种特异性粘附蛋白编码基因pvpA的实时荧光定量PCR检测方法。【方法】根据GenBank公布的不同国家和地区鸡毒支原体pvpA基因序列,在高度保守区域设计一对引物。以临床分离鸡毒支原体RC1为模板扩增pvpA基因,将...【目的】建立基于鸡毒支原体种特异性粘附蛋白编码基因pvpA的实时荧光定量PCR检测方法。【方法】根据GenBank公布的不同国家和地区鸡毒支原体pvpA基因序列,在高度保守区域设计一对引物。以临床分离鸡毒支原体RC1为模板扩增pvpA基因,将其连接到PMD19-T载体,转化至大肠杆菌DH5α中,经PCR和酶切鉴定并测序验证后得到阳性重组质粒rPvpA90。以rPvpA90为模板建立SYBR Green I荧光定量的标准曲线和溶点曲线,并进行特异性,灵敏性,重复性及临床样本检测试验,评价该方法的可行性。【结果】所建立的荧光定量PCR标准曲线循环阈值与模板浓度呈良好的线性关系,溶点曲线特异,相关系数为0.990。最低检测限为72拷贝/20μL,其敏感性比常规PCR至少高100倍;无论是对不同病原DNA单模板还是几种病原DNA混合模板进行扩增,该方法都呈现很好的特异性;重复性试验中,批内和批间变异系数均小于2%,表明该方法重现性好;临床样本的检测结果表明所建立的荧光定量PCR检测方法的检测率明显高于常规PCR方法。【结论】本研究初步建立了基于种特异性基因pvpA的鸡毒支原体荧光定量PCR方法,为养禽场诊断和监测鸡毒支原体病原提供一种新的特异、灵敏的方法。展开更多
基金supported by the National Natural Science Foundation of China(31972681)the National Key Research and Development Program of China(2017YFD0501500)。
文摘Mycoplasma gallisepticum HS strain(MG-HS)is a pathogen that causes chronic respiratory disease(CRD)in chicken,which is characterized by host respiratory inflammatory damage,brings huge economic losses to the poultry industry.Recently,emerging Chinese herbal medicines(CHM)have been used to treat CRD.This study was aimed to investigate the preventive and therapeutic effects and their potential mechanisms of Chinese herbal medicinal formulae(CHMF),which consisted of 10 kinds of Chinese herbal medicine including Scutellaria,Houttuynia cordate and licorice,on MGinduced CRD in chickens.With respect to the preventive effect,the results showed that CHMF could effectively recover the MG-induced decrease on body weight and feed conversion ratio.Histopathological analysis showed that both prevention and treatment of CHMF could significantly alleviate the severe respiratory inflammation induced by MG infection.Moreover,compared with the MG infection group,both the prevention groups and the treatment groups of CHMF could effectively reduce the expression of MG adhesion protein(p MGA1.2)to inhibit the proliferation of MG,and thus effectively inhibit the expression of MG-induced inflammatory factors interleukin-1β(IL-1β)and tumour necrosis factor-α(TNF-α).In summary,these findings confirm that CHMF can protect chickens from various tissue damage caused by MG infection and has no adverse effects on the performance of chickens in the short term.And its efficacy against MG is equal to or better than that of tiamulin.
文摘The study and entire laboratory works were conducted from December 2014 to April 2015 in National Veterinary Institute, Bishoftu, Ethiopia. Formaldehyde inactivated Montanide ISA70 based Mycoplasma gallisepticum (MG) trial vaccine strain was confirmed the identity with known primer using PCR from locally isolates of National Veterinary Institute of Ethiopia. This study was aimed to develop formaldehyde inactivated Montanide ISA70 based MG vaccine in Ethiopia. It can help to device strategies in controlling the disease mainly through developing more effective vaccine which will replace the currently being imported vaccines by some farms. After culturing procedure, oil based inactivated MG trial vaccine was produced in suitable clean and secure accommodation. In this study, among different isolates, local isolate of Samuel farm in NVI was prepared and evaluated in chickens. The amount of immune antigen per 0.5 ml of the dose was 107 Colony forming units (CFU) of the bacteria. The trail vaccine was prepared and evaluated at the age of 16 weeks of chickens;the chickens were randomly divided into three groups (A, B and C), each having twenty birds (10 male and 10 female). Each of group B was vaccinated group of imported-live vaccine with 30 μl intraocularly for comparing with inactivated trial vaccine, each bird of group C was inoculated with 0.5 ml indigenous or trial vaccine subcutaneously at mid neck region and group A was used as a control then challenge tests were performed. After challenge test, among non-vaccinated chickens (control or group A) 2 chickens were died (10%), thicken and cloudy appearance of the air sac showed 18 (90%), 2 chickens were not showed thickened and cloudy air sack (10%). Although among vaccinated group (inactivated vaccine or group C), all chickens did not show clinical signs or post mortem changes (100%). From attenuated imported live vaccine (group B), no clinical signs or post mortem changes were observed (100%). It was concluded that oil based MG vaccine induces protective level of anti MG antibodies in chickens.
基金supported by the National Natural Science Foundation of China (Grant Nos.30871883 and 31001077)the National Basic Foundation of China (Grant Nos.2006JB01 and 2008JB13)the Shanghai Agri & Tech Foundation (Grant No.2007-11-2)
文摘The detailed genetic analysis of mycoplasmas has long been hampered by the lack of appropriate tools for genetic manipulation. In this study, the transposon vector, mini-Tn4001tetM, was constructed containing the tnp gene, encoding a transposase gene in Staphylococcus aureus, two copies of the IS256 inverted repeat sequence (inner and outer) and the tetM gene, from the Enterococcus faecalis Tn916 transposon, conferring resistance to tetracycline. This vector was electro-transformed into Mycoplasma gallisepticum (MG). The recombinant cells were screened by tetracycline selection. The results indicated that the transposon vector could replicate in MG strain R by successive passages, indicating that MG is a potential vector for expressing protective antigens of other pathogens.
文摘【目的】建立基于鸡毒支原体种特异性粘附蛋白编码基因pvpA的实时荧光定量PCR检测方法。【方法】根据GenBank公布的不同国家和地区鸡毒支原体pvpA基因序列,在高度保守区域设计一对引物。以临床分离鸡毒支原体RC1为模板扩增pvpA基因,将其连接到PMD19-T载体,转化至大肠杆菌DH5α中,经PCR和酶切鉴定并测序验证后得到阳性重组质粒rPvpA90。以rPvpA90为模板建立SYBR Green I荧光定量的标准曲线和溶点曲线,并进行特异性,灵敏性,重复性及临床样本检测试验,评价该方法的可行性。【结果】所建立的荧光定量PCR标准曲线循环阈值与模板浓度呈良好的线性关系,溶点曲线特异,相关系数为0.990。最低检测限为72拷贝/20μL,其敏感性比常规PCR至少高100倍;无论是对不同病原DNA单模板还是几种病原DNA混合模板进行扩增,该方法都呈现很好的特异性;重复性试验中,批内和批间变异系数均小于2%,表明该方法重现性好;临床样本的检测结果表明所建立的荧光定量PCR检测方法的检测率明显高于常规PCR方法。【结论】本研究初步建立了基于种特异性基因pvpA的鸡毒支原体荧光定量PCR方法,为养禽场诊断和监测鸡毒支原体病原提供一种新的特异、灵敏的方法。
