[ Objective] To develop multiplex-PCR for diagnosis of mixed infection caused by Pasteurella multocida ( PM), Haomophilus parasuis (HPS) and Actinbaci/lus pleuropneumoniae (App). [ Method ] PCR method was develo...[ Objective] To develop multiplex-PCR for diagnosis of mixed infection caused by Pasteurella multocida ( PM), Haomophilus parasuis (HPS) and Actinbaci/lus pleuropneumoniae (App). [ Method ] PCR method was developed to detect single infection caused by PM, HPS or App. The conditions of amplification and primers were optimized, and the multiple-PCR was developed to detect mixed infection of PM, HPS and App. [ Result] A 457-bp band, a 821-bp band and a 342-bp band were simultaneously amplified in the one PCR reaction system. The method had high sensitivity and specificity. [ Conduslon] The multiple-PCR is successfully developed and can be used for differential diagnosis of PM, HPS and App.展开更多
Pasteurella multocida, a Gram-negative nonmotile coccobacillus, is the causative agent of fowl cholera, bovine hemorrhagic septicemia, enzoonotic pneumonia and swine atropic rhinitis. Two filamentous hemagglutinin gen...Pasteurella multocida, a Gram-negative nonmotile coccobacillus, is the causative agent of fowl cholera, bovine hemorrhagic septicemia, enzoonotic pneumonia and swine atropic rhinitis. Two filamentous hemagglutinin genes, fhaB1 and JhaB2, are the potential virulence factors. In this study, an inactivationfhaB1 mutant ofP. multocida in avian strain C48-102 was constructed by a kanamycin-resistance cassette. The virulence of thefhaB1 mutant and the wild type strain was assessed in chickens by intranasal and intramuscular challenge. The inactivation offhaB1 resulted in a high degree of attenuation when the chickens were challenged intranasally and a lesser degree when challenged intramuscularly. ThefhaB1 mutant and the wild type strain were investigated their sensitivity to the antibody-dependent classical complement-mediated killing pathway in 90% convalescent chicken serum. ThefhaB1 mutant was serum sensitive as the viability has reduced between untreated serum and heat inactivated chicken serum (P〈0.007). These results confirmed that FhaB1 played the critical roles in the bacterial pathogenesis and further studies were needed to investigate the mechanism which caused reduced virulence of the fhaB1 mutant.展开更多
Dear Editor,Rabbit hemorrhagic disease(RHD)is a highly contagious disease of both wild and domesticated rabbits(Oryctolagus cuniculus).The causative agent of the disease is the rabbit hemorrhagic disease virus(RHDV),b...Dear Editor,Rabbit hemorrhagic disease(RHD)is a highly contagious disease of both wild and domesticated rabbits(Oryctolagus cuniculus).The causative agent of the disease is the rabbit hemorrhagic disease virus(RHDV),belongs to the genus Lagovirus within the family Caliciviridae(Granzow et al.,1996;Ohlinger et al.,1990).It is a small and non-enveloped virus with a 7.5 kb single stranded positive sense RNA genome(Meyers et al.,1991;Meyers et al.,2000).Based on an analysis of VP60展开更多
为建立同时检测家兔肺炎克雷伯菌(K.pneum oniae)、兔病毒性出血症病毒(RH D V)、多杀性巴氏杆菌(P.m ultocida)和支气管败血波氏杆菌(B.bronchiseptica)的多重PC R方法,根据G en Bank中肺炎克雷伯菌的16S r RN A基因、RHDV的VP60基因...为建立同时检测家兔肺炎克雷伯菌(K.pneum oniae)、兔病毒性出血症病毒(RH D V)、多杀性巴氏杆菌(P.m ultocida)和支气管败血波氏杆菌(B.bronchiseptica)的多重PC R方法,根据G en Bank中肺炎克雷伯菌的16S r RN A基因、RHDV的VP60基因、多杀性巴氏杆菌的ktm1基因和支气管败血波氏杆菌的ptx A基因的保守序列设计4对特异性引物,优化反应体系与反应条件,建立能同时扩增兔肺炎克雷伯菌(127bp)、RHDV(322 bp)、多杀性巴氏杆菌(457 bp)和支气管败血波氏杆菌(872 bp)相应基因的四重PCR方法。结果显示,该方法能单管同时特异性检测兔源肺炎克雷伯菌、RHDV、多杀性巴氏杆菌与支气管败血波氏杆菌,且对兔源金黄色葡萄球菌、无乳链球菌、大肠杆菌与魏氏梭菌等无特异性扩增,说明该方法具有较强的特异性。该方法的灵敏度分别为9.16、7.24×10-3、7.96、6.8×10-1pg/L。采用建立的方法对人工感染试验病例与临床样本检测结果表明,该方法能快速、特异地鉴别诊断4种病原的单独与混合感染,与单一PCR方法相比,无显著性差异(P>0.05)。该方法的建立为临床上鉴别诊断4种家兔呼吸道疾病病原提供了重要的技术手段。展开更多
P asteurella multocida (P. multocida), a Gram-negative pathogen, is mainly associated with animal infections. Human P. multocida infections are occasionally encountered and most human cases are due to the anima...P asteurella multocida (P. multocida), a Gram-negative pathogen, is mainly associated with animal infections. Human P. multocida infections are occasionally encountered and most human cases are due to the animal exposure or contact, especially due to cat bites. 1 In China, P. multocida infections are rarely described. CASE REPORT A 50-year old woman was sent to the Emergency Department of West China Hospital on June 25, 2004, due to the complain of an onset of pain, erythema, bleeding, and swelling at a wound on the right leg, caused by a pet cat bite 11 hours ago. She was diagnosed as having a wound infection and received intravenous penicillin G 4 million U/day. One day later, though the bleeding was ceased, she felt worse. Then, she was admitted to the Department of Infectious Diseases. She has no fever nor other systemic symptoms when she was admitted. On examination, three close, small and deep holes were found on the right leg near the ankle. Around the holes, the skin and soft tissue were red swelling and tenderness with a high skin temperature. Little pus was presented when the wound was crushed. Regional lymphadenopathy in right groin was found. No dyskinesia and tenderness of the ankle joint was found. Examination of her respiratory, cardiovascular, and gastrointestinal systems was unremarkable. Initial blood routine examination for her revealed: haemoglobin 12.5 g/dl; white blood cell count 7×10 9/L (neutrophils 4.48, lymphocytes 2.45); platelet, 184×10 9/L. But the white cell count increased to 11.2×10 9/L (neutrophils 8.52, lymphocytes 2.58) on the next day. Routine biochemical examination for her revealed normality.展开更多
Respiratory diseases are one of the most important health problems in pig herds. The porcine respiratory disease complex (PRDC) is the term used to describe pneumonic diseases caused by multiple infectious agents that...Respiratory diseases are one of the most important health problems in pig herds. The porcine respiratory disease complex (PRDC) is the term used to describe pneumonic diseases caused by multiple infectious agents that provoke weight loss in animals or death. In the PRDC multiple pathogens (bacteria and/or viruses) work in combination to induce this respiratory disease. Within this complex, Actinobacillus pleuropneumoniae, Streptococcus suis, Pasteurella multocida, Bordetella bronchiseptica, Haemophilus parasuis and Mycoplasma hyopneumoniae are the main bacterial pathogens involved in great economic losses to the swine industry. The aim of this work was to estimate the presence of A. pleuropneumoniae, S. suis, P. multocida, B. bronchiseptica, H. parasuis and M. hyopneumoniae in the upper respiratory tract of pigs in representative swine farms inAguascalientes,Mexico, using PCR technique. The study was performed in 14 swine farms. We obtained a total of 212 nasal swabs. Near 20% of samples were positive for A. pleuropneumoniae (located in the 79% of farms);17% were positive for S. suis (in 86% of farms), of these, 3% were S. suis serovar 2;30% were positive for H. parasuis (93% of farms);23% of the samples to P. multocida (in 79% of farms);and 19% to M. hyopneumoniae (in 64% of farms). B. bronchiseptica was not detected in this study. The results obtained show that bacterial pathogens of PRDC were present in the upper respiratory tract of pigs in all farms studied;therefore, these pathogens are widely disseminated in pig farms of Aguascalientes, Mexico.展开更多
Background:Pasteurella multocida is an important and old zoonotic pathogen worldwide which has an impressive host spectrum including numerous domestic and wild animals as well as birds,causing specific diseases or out...Background:Pasteurella multocida is an important and old zoonotic pathogen worldwide which has an impressive host spectrum including numerous domestic and wild animals as well as birds,causing specific diseases or outbreak with great economic impact.It has never been reported that P.multocida can cause an epidemic in wild rodents.In June 5–17,2016,more than 1000 rodent deaths of an unknown cause quickly spread in the PuEr City,Yunnan province,southwestern China.Methods:The rodents in affected areas and outside of the epidemic areas were collected and screened for possible known pathogens including Yersinia pestis,rabies virus and hantavirus as well as other bacteria.The possible bacterial pathogens were isolated both by culture medium and by mouse inoculation in parallel.The isolates were identified by the Vitek GNI card and PCR assays for 16S rRNA genes.The pathogen strains were selected for whole genome sequencing analysis.Results:A total of 123 rodents were collected from 25 sample sites at affected area,among of which,all 119 dead rodents were negative for the pathogen under consideration except P.multocida,and all four live rodents were negative for P.multocida.In addition,480 rodents collected from other 23 counties outside of the epidemic area in Yunnan were negative for with P.multocida.A total of 14 strains of P.multocida(six directly isolated from the field rodents and eight from the experimental mice that were injected with the organ substrates from the dead rodents)belonged to serogroup A and serogroup F represented by 9 N and 20 N were identified in these epidemic areas.Whole genome sequencing revealed that the serogroup F strain shared 99%similarity to P.multocida Pm70 from chicken,but contained a 50 k bp insertion sequence.The serogroup A strain shared 95%similarity to P.multocida FDAARGOS_385 from a human patient,but contained four large structural differences.Histological abnormalities were identified in the livers,lungs,hearts and brains of the inoculated mice.Conclusions:The simultaneous occu展开更多
Objective: To evaluate the effects of Pasteurella multocida(P. multocida) vaccines on the expression and release of antibodies, interleukin(IL)-6 and IL-12 by serum. Methods: Balb/c mice were immunized with two formal...Objective: To evaluate the effects of Pasteurella multocida(P. multocida) vaccines on the expression and release of antibodies, interleukin(IL)-6 and IL-12 by serum. Methods: Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA(AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA. Results: The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge(P<0.05). The highest release of these cytokines was obtained by P.multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum(P<0.05). Conclusions: Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines.These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.展开更多
Animal bites are frequently encountered in the emergency department(ED). Aortitis leading to mycotic abdominal aortic aneurysm is a rare and potentially deadly complication of Pasteurella multocida(P. multocida) follo...Animal bites are frequently encountered in the emergency department(ED). Aortitis leading to mycotic abdominal aortic aneurysm is a rare and potentially deadly complication of Pasteurella multocida(P. multocida) following an animal bite. We present the case of a 68-year-old male who presented to the ED after falling at home. He complained of weakness and abdominal pain. He was in septic shock and was treated empirically with broad-spectrum antibiotics and intravenous fluids. He reported previous antibiotic treatment of a cellulitis secondary to a cat bite injury to his right thumb four weeks prior. Abdominal ultrasound and subsequent computed tomography scan revealed a leaking mycotic abdominal aneurysm that was surgically repaired. Blood cultures and aortic wall tissue cultures grew P. multocida. Given how common animal bite presentations are in the ED, this case highlights the need to consider aortitis and mycotic abdominal aortic aneurysm in an unwell patient with an animal bite.展开更多
Objective:To characterize Pasteurella isolated from backyard chickens using whole cell protein lysate profiles and random amplified polymorphic DNA(RAPD)techniques to show their genetic relationship because Pasteurcll...Objective:To characterize Pasteurella isolated from backyard chickens using whole cell protein lysate profiles and random amplified polymorphic DNA(RAPD)techniques to show their genetic relationship because Pasteurclla multocida(P.multocida)is an important cause of fatal infections in backyard chickens.Methods:Twenty one P.multocida isolates were recovered previously from clinical cases of fowl cholera belonging to individual owners and phenotypically analysed using biochemical tests and serotyping were used far the genetic characterization.Results:Phylogenetic study based on both methods revealed that the recovered population of P.multocida isolated from backyard chickens differs markedly,constituting a well-separated cluster and appearance of 3 distinguishing lineages with greater discrimination shown by RAPDPCR that resulted in two suclusters in cluster A and three subclusters in cluster B and were related greatly with capsular serogroups for the examined strains.The whole cell protein revealed the presence of dominant protein bands at approximately 41 and 61 kDa in all of the examined isolates that may be a virulent proteins share in the increasing of its pathogenicity.Clear distinctive bands ranged from 123 to 1534 bp.Conclusions:Based on the previous findings,there are three spreading clusters that may indicate the association of a small number of P.multocida variants with the majority of cases suggesting that certain clones of P.multocida are able to colonue the examined backyard chickens.Also,the ease and rapidity of RAPD-PCR support the use of this technique as alternative to the more labour-intensive SDS-PACE system for strain differentiation and epidemiological studies of avian P.multocida.Further application of RAPD technology to the examination of avian cholera outbreaks in commercially available flocks may facilitate more effective management of this disease by providing the potential to investigate correlations of P.multocida genotypes,to identify affiliations between bird types and bacterial genot展开更多
[Objective] To study on genetic inactivation bacterial ghosts of Pasteurella multocida based PhiX174 gene E lysis cassette mediated. [ Method ] Recombinant pPBA1100-e was constructed by which the gene E of bacteriopha...[Objective] To study on genetic inactivation bacterial ghosts of Pasteurella multocida based PhiX174 gene E lysis cassette mediated. [ Method ] Recombinant pPBA1100-e was constructed by which the gene E of bacteriophage Phix174 and temperature sensitivity expressing control system hybridized with plasmid pPBA1100 by genetic engineering method. Recombinant was transformed to Pasteurella multocida and lysis gene E expressed by temperature induction. Recombinant was detected by restriction endonuclease. Cell morphology of bacterial ghost of Pasteurella mul- tocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysis. I Result~ The results indicated that the recombination plasmid presented three bands by restriction endonuclease and agarose electrophoresis and that molecular weight of every band ac- corded with theoretical value. The result of SEM observing showed that recombination plasmid expressed successfully in P. multocida and produced bacterial ghost. The result of CFU detecting demonstrated that inactivation ratio of P. multocida reached 99 per cent. ~Conclusion~ This study pro- vided technical bases for the preparation of antigen vaccine of natural bacterial outer membrane protein.展开更多
基金funded by the subproject of National Key Technology R&D Program (2006BAD06A01 and 2006BAD06A12)Basic Work of National Science and Technology Special Plan (2008FY210200)
文摘[ Objective] To develop multiplex-PCR for diagnosis of mixed infection caused by Pasteurella multocida ( PM), Haomophilus parasuis (HPS) and Actinbaci/lus pleuropneumoniae (App). [ Method ] PCR method was developed to detect single infection caused by PM, HPS or App. The conditions of amplification and primers were optimized, and the multiple-PCR was developed to detect mixed infection of PM, HPS and App. [ Result] A 457-bp band, a 821-bp band and a 342-bp band were simultaneously amplified in the one PCR reaction system. The method had high sensitivity and specificity. [ Conduslon] The multiple-PCR is successfully developed and can be used for differential diagnosis of PM, HPS and App.
基金supported by the National Natural Science Foundation of China(31302109)
文摘Pasteurella multocida, a Gram-negative nonmotile coccobacillus, is the causative agent of fowl cholera, bovine hemorrhagic septicemia, enzoonotic pneumonia and swine atropic rhinitis. Two filamentous hemagglutinin genes, fhaB1 and JhaB2, are the potential virulence factors. In this study, an inactivationfhaB1 mutant ofP. multocida in avian strain C48-102 was constructed by a kanamycin-resistance cassette. The virulence of thefhaB1 mutant and the wild type strain was assessed in chickens by intranasal and intramuscular challenge. The inactivation offhaB1 resulted in a high degree of attenuation when the chickens were challenged intranasally and a lesser degree when challenged intramuscularly. ThefhaB1 mutant and the wild type strain were investigated their sensitivity to the antibody-dependent classical complement-mediated killing pathway in 90% convalescent chicken serum. ThefhaB1 mutant was serum sensitive as the viability has reduced between untreated serum and heat inactivated chicken serum (P〈0.007). These results confirmed that FhaB1 played the critical roles in the bacterial pathogenesis and further studies were needed to investigate the mechanism which caused reduced virulence of the fhaB1 mutant.
基金supported by the Shaanxi Agriculture Science and Technology Research Projects (2014K02-06-01)
文摘Dear Editor,Rabbit hemorrhagic disease(RHD)is a highly contagious disease of both wild and domesticated rabbits(Oryctolagus cuniculus).The causative agent of the disease is the rabbit hemorrhagic disease virus(RHDV),belongs to the genus Lagovirus within the family Caliciviridae(Granzow et al.,1996;Ohlinger et al.,1990).It is a small and non-enveloped virus with a 7.5 kb single stranded positive sense RNA genome(Meyers et al.,1991;Meyers et al.,2000).Based on an analysis of VP60
文摘为建立同时检测家兔肺炎克雷伯菌(K.pneum oniae)、兔病毒性出血症病毒(RH D V)、多杀性巴氏杆菌(P.m ultocida)和支气管败血波氏杆菌(B.bronchiseptica)的多重PC R方法,根据G en Bank中肺炎克雷伯菌的16S r RN A基因、RHDV的VP60基因、多杀性巴氏杆菌的ktm1基因和支气管败血波氏杆菌的ptx A基因的保守序列设计4对特异性引物,优化反应体系与反应条件,建立能同时扩增兔肺炎克雷伯菌(127bp)、RHDV(322 bp)、多杀性巴氏杆菌(457 bp)和支气管败血波氏杆菌(872 bp)相应基因的四重PCR方法。结果显示,该方法能单管同时特异性检测兔源肺炎克雷伯菌、RHDV、多杀性巴氏杆菌与支气管败血波氏杆菌,且对兔源金黄色葡萄球菌、无乳链球菌、大肠杆菌与魏氏梭菌等无特异性扩增,说明该方法具有较强的特异性。该方法的灵敏度分别为9.16、7.24×10-3、7.96、6.8×10-1pg/L。采用建立的方法对人工感染试验病例与临床样本检测结果表明,该方法能快速、特异地鉴别诊断4种病原的单独与混合感染,与单一PCR方法相比,无显著性差异(P>0.05)。该方法的建立为临床上鉴别诊断4种家兔呼吸道疾病病原提供了重要的技术手段。
文摘P asteurella multocida (P. multocida), a Gram-negative pathogen, is mainly associated with animal infections. Human P. multocida infections are occasionally encountered and most human cases are due to the animal exposure or contact, especially due to cat bites. 1 In China, P. multocida infections are rarely described. CASE REPORT A 50-year old woman was sent to the Emergency Department of West China Hospital on June 25, 2004, due to the complain of an onset of pain, erythema, bleeding, and swelling at a wound on the right leg, caused by a pet cat bite 11 hours ago. She was diagnosed as having a wound infection and received intravenous penicillin G 4 million U/day. One day later, though the bleeding was ceased, she felt worse. Then, she was admitted to the Department of Infectious Diseases. She has no fever nor other systemic symptoms when she was admitted. On examination, three close, small and deep holes were found on the right leg near the ankle. Around the holes, the skin and soft tissue were red swelling and tenderness with a high skin temperature. Little pus was presented when the wound was crushed. Regional lymphadenopathy in right groin was found. No dyskinesia and tenderness of the ankle joint was found. Examination of her respiratory, cardiovascular, and gastrointestinal systems was unremarkable. Initial blood routine examination for her revealed: haemoglobin 12.5 g/dl; white blood cell count 7×10 9/L (neutrophils 4.48, lymphocytes 2.45); platelet, 184×10 9/L. But the white cell count increased to 11.2×10 9/L (neutrophils 8.52, lymphocytes 2.58) on the next day. Routine biochemical examination for her revealed normality.
文摘Respiratory diseases are one of the most important health problems in pig herds. The porcine respiratory disease complex (PRDC) is the term used to describe pneumonic diseases caused by multiple infectious agents that provoke weight loss in animals or death. In the PRDC multiple pathogens (bacteria and/or viruses) work in combination to induce this respiratory disease. Within this complex, Actinobacillus pleuropneumoniae, Streptococcus suis, Pasteurella multocida, Bordetella bronchiseptica, Haemophilus parasuis and Mycoplasma hyopneumoniae are the main bacterial pathogens involved in great economic losses to the swine industry. The aim of this work was to estimate the presence of A. pleuropneumoniae, S. suis, P. multocida, B. bronchiseptica, H. parasuis and M. hyopneumoniae in the upper respiratory tract of pigs in representative swine farms inAguascalientes,Mexico, using PCR technique. The study was performed in 14 swine farms. We obtained a total of 212 nasal swabs. Near 20% of samples were positive for A. pleuropneumoniae (located in the 79% of farms);17% were positive for S. suis (in 86% of farms), of these, 3% were S. suis serovar 2;30% were positive for H. parasuis (93% of farms);23% of the samples to P. multocida (in 79% of farms);and 19% to M. hyopneumoniae (in 64% of farms). B. bronchiseptica was not detected in this study. The results obtained show that bacterial pathogens of PRDC were present in the upper respiratory tract of pigs in all farms studied;therefore, these pathogens are widely disseminated in pig farms of Aguascalientes, Mexico.
基金This study was funded by the State Key Research Development Program of China(2016YFC1201902,2016YFC1200301)the Natural Science Foundation of China(81621005,81773492,81760607,81360413)the Program of Cultivation of Technologically Innovative Talents of Yunnan(2014HB093).
文摘Background:Pasteurella multocida is an important and old zoonotic pathogen worldwide which has an impressive host spectrum including numerous domestic and wild animals as well as birds,causing specific diseases or outbreak with great economic impact.It has never been reported that P.multocida can cause an epidemic in wild rodents.In June 5–17,2016,more than 1000 rodent deaths of an unknown cause quickly spread in the PuEr City,Yunnan province,southwestern China.Methods:The rodents in affected areas and outside of the epidemic areas were collected and screened for possible known pathogens including Yersinia pestis,rabies virus and hantavirus as well as other bacteria.The possible bacterial pathogens were isolated both by culture medium and by mouse inoculation in parallel.The isolates were identified by the Vitek GNI card and PCR assays for 16S rRNA genes.The pathogen strains were selected for whole genome sequencing analysis.Results:A total of 123 rodents were collected from 25 sample sites at affected area,among of which,all 119 dead rodents were negative for the pathogen under consideration except P.multocida,and all four live rodents were negative for P.multocida.In addition,480 rodents collected from other 23 counties outside of the epidemic area in Yunnan were negative for with P.multocida.A total of 14 strains of P.multocida(six directly isolated from the field rodents and eight from the experimental mice that were injected with the organ substrates from the dead rodents)belonged to serogroup A and serogroup F represented by 9 N and 20 N were identified in these epidemic areas.Whole genome sequencing revealed that the serogroup F strain shared 99%similarity to P.multocida Pm70 from chicken,but contained a 50 k bp insertion sequence.The serogroup A strain shared 95%similarity to P.multocida FDAARGOS_385 from a human patient,but contained four large structural differences.Histological abnormalities were identified in the livers,lungs,hearts and brains of the inoculated mice.Conclusions:The simultaneous occu
基金part of the project:Study on immune response pattern of cattle vaccinated by Razi pasteurellosis vaccine(Project number:12-18-18-9458-94014)
文摘Objective: To evaluate the effects of Pasteurella multocida(P. multocida) vaccines on the expression and release of antibodies, interleukin(IL)-6 and IL-12 by serum. Methods: Balb/c mice were immunized with two formalin and iron inactivated vaccine doses within 2 weeks. The vaccines were adjuvant with P. multocida A strain, P. multocida B strain and Salmonella typhimurium bacterial DNA(AbDNA, BbDNA and SbDNA for short, respectively). The animals were challenged 4 weeks after immunization. Blood of mice was collected to detect the change of specific antibody, IL-6, and IL-12 using ELISA. Results: The specific antibody and interleukins in the immunized group increased significantly compared to the control mice after vaccination and challenge(P<0.05). The highest release of these cytokines was obtained by P.multocida inactivated with iron and adjuvant with AbDNA at a concentration of 25 μg/mL. The antibody titer peak was 0.447 in mice vaccinated with iron-killed whole-cell antigen adjunct with AbDNA. The time-courses of release showed that bacterial DNA was able to stimulate IL-6 and IL-12 production more than alum(P<0.05). Conclusions: Our findings introduce that bacterial DNA is capable of releasing an immunological response with several cytokines.These indicate that bacterial DNA entrapped with killed P. multocida antigen is a new and effective adjuvant to enhance specific immunity and resistance of animal against the infectious pathogen, which could simplify the development of highly promising strong adjuvant.
文摘Animal bites are frequently encountered in the emergency department(ED). Aortitis leading to mycotic abdominal aortic aneurysm is a rare and potentially deadly complication of Pasteurella multocida(P. multocida) following an animal bite. We present the case of a 68-year-old male who presented to the ED after falling at home. He complained of weakness and abdominal pain. He was in septic shock and was treated empirically with broad-spectrum antibiotics and intravenous fluids. He reported previous antibiotic treatment of a cellulitis secondary to a cat bite injury to his right thumb four weeks prior. Abdominal ultrasound and subsequent computed tomography scan revealed a leaking mycotic abdominal aneurysm that was surgically repaired. Blood cultures and aortic wall tissue cultures grew P. multocida. Given how common animal bite presentations are in the ED, this case highlights the need to consider aortitis and mycotic abdominal aortic aneurysm in an unwell patient with an animal bite.
基金Supported by Regular goveromental annual fund every Fiscal year from Assiut university(Grant No.9/020/08595/0)
文摘Objective:To characterize Pasteurella isolated from backyard chickens using whole cell protein lysate profiles and random amplified polymorphic DNA(RAPD)techniques to show their genetic relationship because Pasteurclla multocida(P.multocida)is an important cause of fatal infections in backyard chickens.Methods:Twenty one P.multocida isolates were recovered previously from clinical cases of fowl cholera belonging to individual owners and phenotypically analysed using biochemical tests and serotyping were used far the genetic characterization.Results:Phylogenetic study based on both methods revealed that the recovered population of P.multocida isolated from backyard chickens differs markedly,constituting a well-separated cluster and appearance of 3 distinguishing lineages with greater discrimination shown by RAPDPCR that resulted in two suclusters in cluster A and three subclusters in cluster B and were related greatly with capsular serogroups for the examined strains.The whole cell protein revealed the presence of dominant protein bands at approximately 41 and 61 kDa in all of the examined isolates that may be a virulent proteins share in the increasing of its pathogenicity.Clear distinctive bands ranged from 123 to 1534 bp.Conclusions:Based on the previous findings,there are three spreading clusters that may indicate the association of a small number of P.multocida variants with the majority of cases suggesting that certain clones of P.multocida are able to colonue the examined backyard chickens.Also,the ease and rapidity of RAPD-PCR support the use of this technique as alternative to the more labour-intensive SDS-PACE system for strain differentiation and epidemiological studies of avian P.multocida.Further application of RAPD technology to the examination of avian cholera outbreaks in commercially available flocks may facilitate more effective management of this disease by providing the potential to investigate correlations of P.multocida genotypes,to identify affiliations between bird types and bacterial genot
基金Supported by the Hongkong Fok Ying Tung Ming Yuan Fund(HK314-14591)Guangdong Provincial Agricultural Research Projects(2004B20201019)Shaoguan Science and Technology Innovation Projects(2010140473)
文摘[Objective] To study on genetic inactivation bacterial ghosts of Pasteurella multocida based PhiX174 gene E lysis cassette mediated. [ Method ] Recombinant pPBA1100-e was constructed by which the gene E of bacteriophage Phix174 and temperature sensitivity expressing control system hybridized with plasmid pPBA1100 by genetic engineering method. Recombinant was transformed to Pasteurella multocida and lysis gene E expressed by temperature induction. Recombinant was detected by restriction endonuclease. Cell morphology of bacterial ghost of Pasteurella mul- tocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysis. I Result~ The results indicated that the recombination plasmid presented three bands by restriction endonuclease and agarose electrophoresis and that molecular weight of every band ac- corded with theoretical value. The result of SEM observing showed that recombination plasmid expressed successfully in P. multocida and produced bacterial ghost. The result of CFU detecting demonstrated that inactivation ratio of P. multocida reached 99 per cent. ~Conclusion~ This study pro- vided technical bases for the preparation of antigen vaccine of natural bacterial outer membrane protein.