The Experts below are selected from a list of 447 Experts worldwide ranked by ideXlab platform
Yutaka Tamura - One of the best experts on this subject based on the ideXlab platform.
-
phylogenetic analysis and pcr detection of Clostridium chauvoei Clostridium haemolyticum Clostridium novyi types a and b and Clostridium septicum based on the flagellin gene
Veterinary Microbiology, 2002Co-Authors: Yoshimasa Sasaki, Noriyasu Takikawa, Akemi Kojima, Hiroshi Aoki, Yasuaki Ogikubo, Yutaka TamuraAbstract:Abstract The flagellin genes ( fliC ) of Clostridium chauvoei , Clostridium haemolyticum , Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene ( fliC ) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I ( Collins et al., 1994 ), are more closely related to Bacillus subtilis than to Clostridium difficile , which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei , C. haemolyticum , C. novyi types A and B, and C. septicum . PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
-
phylogenetic analysis and pcr detection of Clostridium chauvoei Clostridium haemolyticum Clostridium novyi types a and b and Clostridium septicum based on the flagellin gene
Veterinary Microbiology, 2002Co-Authors: Yoshimasa Sasaki, Noriyasu Takikawa, Akemi Kojima, Hiroshi Aoki, Yasuaki Ogikubo, Yutaka TamuraAbstract:The flagellin genes (fliC) of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene (fliC) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I (), are more closely related to Bacillus subtilis than to Clostridium difficile, which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei, C. haemolyticum, C. novyi types A and B, and C. septicum. PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
-
amplification of the 16s 23s rdna spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum
Research in Veterinary Science, 2001Co-Authors: Y Sasaki, Yasuaki Ogikubo, Kinya Yamamoto, Mari Norimatsu, K Amimoto, A Kojima, H Ogata, Yutaka TamuraAbstract:Amplification of the 16S-23S rDNA spacer region by polymerase chain reaction (PCR) was used for the rapid detection of Clostridium chauvoei and C septicum. To assess its specificity, PCR was performed with total DNA from 42 strains of clostridia and three strains of other genera. PCR products specific to C chauvoei or to C septicum were generated from homologous cultures only. Clostridium chauvoer-specific or C septicum-specific amplicons were also generated from tissues of cows experimentally infected with C chauvoei or C septicum and in DNA samples from cows clinically diagnosed as having blackleg or malignant oedema. These results suggest that a species-specific PCR may be useful for the rapid and direct detection of C chauvoei and C septicum in clinical specimens.
-
amplification of the 16s 23s r dna spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum
Research in Veterinary Science, 2001Co-Authors: Y Sasaki, Yasuaki Ogikubo, Kinya Yamamoto, Mari Norimatsu, K Amimoto, A Kojima, H Ogata, Yutaka TamuraAbstract:Abstract Amplification of the 16S-23S r DNA spacer region by polymerase chain reaction ( PCR ) was used for the rapid detection of Clostridium chauvoei and C septicum . To assess its specificity, PCR was performed with total DNA from 42 strains of clostridia and three strains of other genera. PCR products specific to C chauvoei or to C septicum were generated from homologous cultures only. Clostridium chauvoer -specific or C septicum -specific amplicons were also generated from tissues of cows experimentally infected with C chauvoei or C septicum and in DNA samples from cows clinically diagnosed as having blackleg or malignant oedema. These results suggest that a species-specific PCR may be useful for the rapid and direct detection of C chauvoei and C septicum in clinical specimens.
-
rapid detection and identification of Clostridium chauvoei by pcr based on flagellin gene sequence
Veterinary Microbiology, 2001Co-Authors: Akemi Kojima, Yoshimasa Sasaki, Yasuaki Ogikubo, Ikuo Uchida, Tsutomu Sekizaki, Yutaka TamuraAbstract:We developed a one-step polymerase chain reaction (PCR) system that specifically detects Clostridium chauvoei. Oligonucleotide primers were designed to amplify a 516-bp fragment of the structural flagellin gene. The specificity of the PCR was investigated by analyzing 59 strains of clostridia, and seven strain of other genera. A 516-bp fragment could be amplified from all the C. chauvoei strains tested, and no amplification was observed by using DNAs from the other strains tested, including Clostridium septicum. Similarly, this PCR-based method specifically detected C. chauvoei DNA sequences in samples of muscle and exudate of obtained from mice within 12 h of inoculation. In tests using samples of muscle or liver, the limit of detection was about 200 organisms per reaction. These results suggest that the one-step PCR system may be useful for direct detection and identification of C. chauvoei in clinical specimens.
Francisco Carlos Faria Lobato - One of the best experts on this subject based on the ideXlab platform.
-
intracellular survival of Clostridium chauvoei in bovine macrophages
Veterinary Microbiology, 2017Co-Authors: Prhiscylla Sadanã Pires, Rodrigo Otávio Silveira Silva, Renato L Santos, Tatiane A Paixao, Laura Cristina Oliveira Bernardes, Auricelio A Macedo, Luciana Aramuni Goncalves, Carlos Augusto Oliveira, Francisco Carlos Faria LobatoAbstract:Clostridium chauvoei is the etiological agent of blackleg, a severe disease of domestic ruminants, causing myonecrosis and serious toxemia with high mortality. Despite the known importance of this agent, studies evaluating its pathogenesis of blackleg are scarce, and many are based on an unproven hypothesis that states that macrophages are responsible for carrying C. chauvoei spores from the intestines to muscles in the early stages of blackleg. Therefore, the present study aimed to investigate the survival of C. chauvoei vegetative cells or spores after phagocytosis by a murine macrophage cell line (RAW 264.7) and bovine monocyte-derived macrophages and to profile inflammatory and anti-inflammatory cytokine transcripts of bovine macrophages infected with C. chauvoei vegetative cells or spores. Both vegetative cells and spores of C. chauvoei remain viable after internalization by murine and bovine macrophages. Bovine macrophages infected with vegetative cells showed a pro-inflammatory profile, while those infected with spores displayed an anti-inflammatory profile. Together, these results corroborate the classical hypothesis that macrophages may play a role in the early pathogenesis of blackleg. Moreover, this is the first study to evaluate the infection kinetics and cytokine profile of bovine monocyte-derived macrophages infected with a Clostridium species.
-
edema maligno em equino causado por Clostridium chauvoei malignant edema caused by Clostridium chauvoei in a horse
2013Co-Authors: Juliana Targino, Silva Almeida, Emmanuel Emydio, G Pinheiro, Ricardo Santana De Oliveira, Rodrigo Otavio, S A Silva, Francisco Carlos Faria Lobato, Pedro Miguel Ocampos PedrosoAbstract:Background: Malignant edema occurs when a wound is contaminated by one or a combination of the following bacteria: Clostridium septicum, C. sordellii, C. chauvoei, C. novyi type A, and C. perfringens type A. Although malignant edema has been recognized as a disease of economic importance in domestic ruminants, it has rarely been reported in horses in Brazil. Immunohistochemistry is a method that combines histological, immunological and biochemical techniques which, with the use of specifi c antibodies, detects the agent in situ. This paper reports a case of malignant edema by Clostridium chauvoei in a horse in Bahia, Brazil. Case: A 4.5-year-old male horse was submitted for necropsy. The clinical history was obtained from the owner. Samples from the abdominal cavity, thoracic cavity, central nervous system and integumentary system were collected. The samples were fi xed in 10% formalin, processed routinely for histology, embedded in paraffi n, sectioned at fi microns, and stained with hematoxylin and eosin. Additionally, histological sections were subjected to immunohistochemistry anti-Clostridium perfringens type A, C. septicum, C. chauvoei, C. novyi type A, and C. sordellii. Positive and negative controls were included. The animal presented diffi culty in walking, swelling of the pelvic limbs two days after intramuscular anti-infl ammatory treatment, and died seven hours after the onset of clinical signs. Macroscopically, large amounts of hemorrhagic edema and tissue emphysema in the subcutaneous tissue and skeletal muscles were seen. Microscopically, there was moderate bleeding, particularly in subcutaneous areas and between myofi ber bundles, with few and discrete degeneration foci. Immunohistochemistry technique was positive for Clostridium chauvoei. Discussion: The diagnosis of malignant edema was confi rmed considering clinical and pathological alterations as well as the positive immunohistochemistry result for Clostridium chauvoei. Cases of malignant edema are occasionally observed in horses, and Clostridium perfringens and Clostridium septicum are the major etiological agents involved. Two cases have been reported in Brazil, both caused by Clostridium septicum. As in the present report, the animals developed the disease after administration of intramuscular anti-infl ammatory medication. The clinical course of the disease in horses is considered severe, with the fi rst clinical signs occurring between 24 and 48 h post infection. Clinical signs, particularly the marked diffi culty in walking, edema, and crepitus of the affected areas, may be considered characteristic. The pathological fi ndings were similar to those described by other authors, which are attributed to toxins produced by the bacteria; however, the lesions described in this study were similar to those reported in less severe infections by Clostridium septicum. Bacterial culture and polymerase chain reaction (PCR) are standards for the identifi cation of Clostridium species; however, immunohistochemistry is considered a very sensitive and specifi c diagnostic technique. To control the disease, animals may be treated with high doses of penicillin or broad-spectrum antibiotics, which are effi cient when used during the early stages of the disease. Prophylactic measures include good hygiene while handling syringes and surgical instruments, as well as annual vaccination of the animals.
-
Edema maligno em equino causado por Clostridium chauvoei
Acta Scientiae Veterinariae, 2013Co-Authors: Juliana Targino Silva Almeida E Macêdo, Ricardo Santana De Oliveira, Francisco Carlos Faria Lobato, Prhiscylla Sadanã Pires, Emmanuel Emydio Gomes Pinheiro, Rodrigo Otávio Silveira Silva, Pedro Miguel Ocampos PedrosoAbstract:Background: Malignant edema occurs when a wound is contaminated by one or a combination of the following bacteria: Clostridium septicum, C. sordellii, C. chauvoei, C. novyi type A, and C. perfringens type A. Although malignant edema has been recognized as a disease of economic importance in domestic ruminants, it has rarely been reported in horses in Brazil. Immunohistochemistry is a method that combines histological, immunological and biochemical techniques which, with the use of specifi c antibodies, detects the agent in situ. This paper reports a case of malignant edema by Clostridium chauvoei in a horse in Bahia, Brazil. Case: A 4.5-year-old male horse was submitted for necropsy. The clinical history was obtained from the owner. Samples from the abdominal cavity, thoracic cavity, central nervous system and integumentary system were collected. The samples were fi xed in 10% formalin, processed routinely for histology, embedded in paraffi n, sectioned at fi ve microns, and stained with hematoxylin and eosin. Additionally, histological sections were subjected to immunohistochemistry anti-Clostridium perfringens type A, C. septicum, C. chauvoei, C. novyi type A, and C. sordellii. Positive and negative controls were included. The animal presented diffi culty in walking, swelling of the pelvic limbs two days after intramuscular anti-infl ammatory treatment, and died seven hours after the onset of clinical signs. Macroscopically, large amounts of hemorrhagic edema and tissue emphysema in the subcutaneous tissue and skeletal muscles were seen. Microscopically, there was moderate bleeding, particularly in subcutaneous areas and between myofi ber bundles, with few and discrete degeneration foci. Immunohistochemistry technique was positive for Clostridium chauvoei. Discussion: The diagnosis of malignant edema was confi rmed considering clinical and pathological alterations as well as the positive immunohistochemistry result for Clostridium chauvoei. Cases of malignant edema are occasionally observed in horses, and Clostridium perfringens and Clostridium septicum are the major etiological agents involved. Two cases have been reported in Brazil, both caused by Clostridium septicum. As in the present report, the animals developed the disease after administration of intramuscular anti-infl ammatory medication. The clinical course of the disease in horses is considered severe, with the fi rst clinical signs occurring between 24 and 48 h post infection. Clinical signs, particularly the marked diffi culty in walking, edema, and crepitus of the affected areas, may be considered characteristic. The pathological findings were similar to those described by other authors, which are attributed to toxins produced by the bacteria; however, the lesions described in this study were similar to those reported in less severe infections by Clostridium septicum. Bacterial culture and polymerase chain reaction (PCR) are standards for the identifi cation of Clostridium species; however, immunohistochemistry is considered a very sensitive and specifi c diagnostic technique. To control the disease, animals may be treated with high doses of penicillin or broad-spectrum antibiotics, which are effi cient when used during the early stages of the disease. Prophylactic measures include good hygiene while handling syringes and surgical instruments, as well as annual vaccination of the animals.
-
pcr multiplex para identificacao de isolados de Clostridium chauvoei e Clostridium septicum
Arquivo Brasileiro De Medicina Veterinaria E Zootecnia, 2008Co-Authors: Ronnie Antunes De ,assis, Francisco Carlos Faria Lobato, Z.i.p. Lobato, M.f. Camargos, R.a.p. Nascimento, A.p.c. Vargas, F.m. Salvarani, Francisco A UzalAbstract:Multiplex PCR was optimized to detect Clostridium chauvoei and Clostridium septicum in pure cultures. In each reaction, a pair of primers for a specific segment of the flagellin gene of C. chauvoei and a pair of primers for a specific segment of alpha toxin gene of C. septicum were employed. Reference strains of both microorganisms were used as control. The multiplex PCR was evaluated by testing 16 clinical isolates of C. chauvoei from ruminants, 15 clinical isolates of C. septicum from ruminants and, four vaccine strains of each one of these agents. Reference strains of both microorganisms were used as control. To evaluate the specificity, genomic DNA of the following microorganisms was used: C. sordellii, C. novyi type A, C. novyi type B, C. perfringens type A, C. haemolyticum, C. botulinum type D, Pseudomonas aeruginosa, Staphylococcus aureus, Enterobacter aerogenes, Escherichia coli, and Salmonella typhimurium. All the isolates and vaccine strains of C. chauvoei and C. septicum were positive by PCR assay and cross reactions were not observed with the other species of clostridia, the other bacterial species or amongst both investigated agents. The smallest concentrations of DNA detected from C. chauvoei and C. septicum were 45pg/µl and 30pg/µl, respectively. The multiplex PCR was useful for the specific identification of C. chauvoei and C. septicum in pure cultures.
-
reacao em cadeia da polimerase para deteccao de Clostridium chauvoei em tecidos de cavia porcellus
Pesquisa Agropecuaria Brasileira, 2005Co-Authors: Ronnie Antunes De ,assis, Francisco Carlos Faria Lobato, Mariano Fernandez E Miyakawa, Francisco Alejandro UzalAbstract:The objective of this work was the standardization of a polymerase chain reaction (PCR) for detection of Clostridium chauvoei in formalin-fixed, paraffin-embedded tissues of guinea-pigs (Cavia porcellus) infected experimentally with this microorganism. The animals were sacrificed, and samples of muscle from inoculation area (MIA), liver, myocardium and spleen were available for PCR technique. Clostridium chauvoei was detected in all sections of the MIA, liver and myocardium, and no product was observed in sections of the spleen. Cross-reactions were not observed in sections of MIA of the animals inoculated with other clostridia, as well as no amplification was observed in sections of MIA of control animals. These results show that the PCR technique developed in this study may be useful for detection of C. chauvoei in formalin-fixed, paraffin-embedded tissues.
Ronnie Antunes De ,assis - One of the best experts on this subject based on the ideXlab platform.
-
pcr multiplex para identificacao de isolados de Clostridium chauvoei e Clostridium septicum
Arquivo Brasileiro De Medicina Veterinaria E Zootecnia, 2008Co-Authors: Ronnie Antunes De ,assis, Francisco Carlos Faria Lobato, Z.i.p. Lobato, M.f. Camargos, R.a.p. Nascimento, A.p.c. Vargas, F.m. Salvarani, Francisco A UzalAbstract:Multiplex PCR was optimized to detect Clostridium chauvoei and Clostridium septicum in pure cultures. In each reaction, a pair of primers for a specific segment of the flagellin gene of C. chauvoei and a pair of primers for a specific segment of alpha toxin gene of C. septicum were employed. Reference strains of both microorganisms were used as control. The multiplex PCR was evaluated by testing 16 clinical isolates of C. chauvoei from ruminants, 15 clinical isolates of C. septicum from ruminants and, four vaccine strains of each one of these agents. Reference strains of both microorganisms were used as control. To evaluate the specificity, genomic DNA of the following microorganisms was used: C. sordellii, C. novyi type A, C. novyi type B, C. perfringens type A, C. haemolyticum, C. botulinum type D, Pseudomonas aeruginosa, Staphylococcus aureus, Enterobacter aerogenes, Escherichia coli, and Salmonella typhimurium. All the isolates and vaccine strains of C. chauvoei and C. septicum were positive by PCR assay and cross reactions were not observed with the other species of clostridia, the other bacterial species or amongst both investigated agents. The smallest concentrations of DNA detected from C. chauvoei and C. septicum were 45pg/µl and 30pg/µl, respectively. The multiplex PCR was useful for the specific identification of C. chauvoei and C. septicum in pure cultures.
-
reacao em cadeia da polimerase para deteccao de Clostridium chauvoei em tecidos de cavia porcellus
Pesquisa Agropecuaria Brasileira, 2005Co-Authors: Ronnie Antunes De ,assis, Francisco Carlos Faria Lobato, Mariano Fernandez E Miyakawa, Francisco Alejandro UzalAbstract:The objective of this work was the standardization of a polymerase chain reaction (PCR) for detection of Clostridium chauvoei in formalin-fixed, paraffin-embedded tissues of guinea-pigs (Cavia porcellus) infected experimentally with this microorganism. The animals were sacrificed, and samples of muscle from inoculation area (MIA), liver, myocardium and spleen were available for PCR technique. Clostridium chauvoei was detected in all sections of the MIA, liver and myocardium, and no product was observed in sections of the spleen. Cross-reactions were not observed in sections of MIA of the animals inoculated with other clostridia, as well as no amplification was observed in sections of MIA of control animals. These results show that the PCR technique developed in this study may be useful for detection of C. chauvoei in formalin-fixed, paraffin-embedded tissues.
-
Reação em cadeia da polimerase para detecção de Clostridium chauvoei em tecidos de Cavia porcellus
Pesquisa Agropecuária Brasileira, 2005Co-Authors: Ronnie Antunes De ,assis, Lobato, Francisco Carlos Faria, Miyakawa, Mariano Fernandez, Uzal, Francisco AlejandroAbstract:O objetivo deste trabalho foi padronizar uma técnica de reação em cadeia da polimerase (PCR), para detecção de Clostridium chauvoei, em tecidos fixados em formol e incluídos em parafina de cobaias (Cavia porcellus) experimentalmente infectadas com esse microrganismo. Os animais foram sacrificados, e amostras do músculo da área de inoculação (MAI), fígado, miocárdio e baço foram disponibilizadas para a técnica de PCR. O clostrídio foi detectado em todas as secções do MAI, fígado e miocárdio, mas não foi observado em secções do baço. Reações cruzadas não foram observadas a partir de secções do MAI dos animais com inoculação de outras espécies de clostrídios, bem como nenhuma amplificação foi observada a partir de secções do MAI dos animais controle. Esses resultados mostram que a técnica de PCR desenvolvida neste estudo, pode ser usada para detecção de Clostridium chauvoei em tecidos fixados em formol e incluídos em parafina
-
PCR detection of Clostridium chauvoei in tissues of Cavia porcellus
'Embrapa Agroindustria Tropical', 2005Co-Authors: Ronnie Antunes De ,assis, Lobato, Francisco Carlos Faria, Miyakawa, Mariano Fernandez, Uzal, Francisco AlejandroAbstract:O objetivo deste trabalho foi padronizar uma técnica de reação em cadeia da polimerase (PCR), para detecção de Clostridium chauvoei, em tecidos fixados em formol e incluídos em parafina de cobaias (Cavia porcellus) experimentalmente infectadas com esse microrganismo. Os animais foram sacrificados, e amostras do músculo da área de inoculação (MAI), fígado, miocárdio e baço foram disponibilizadas para a técnica de PCR. O clostrídio foi detectado em todas as secções do MAI, fígado e miocárdio, mas não foi observado em secções do baço. Reações cruzadas não foram observadas a partir de secções do MAI dos animais com inoculação de outras espécies de clostrídios, bem como nenhuma amplificação foi observada a partir de secções do MAI dos animais controle. Esses resultados mostram que a técnica de PCR desenvolvida neste estudo, pode ser usada para detecção de Clostridium chauvoei em tecidos fixados em formol e incluídos em parafina.The objective of this work was the standardization of a polymerase chain reaction (PCR) for detection of Clostridium chauvoei in formalin-fixed, paraffin-embedded tissues of guinea-pigs (Cavia porcellus) infected experimentally with this microorganism. The animals were sacrificed, and samples of muscle from inoculation area (MIA), liver, myocardium and spleen were available for PCR technique. Clostridium chauvoei was detected in all sections of the MIA, liver and myocardium, and no product was observed in sections of the spleen. Cross-reactions were not observed in sections of MIA of the animals inoculated with other clostridia, as well as no amplification was observed in sections of MIA of control animals. These results show that the PCR technique developed in this study may be useful for detection of C. chauvoei in formalin-fixed, paraffin-embedded tissues
-
Reação em cadeia da polimerase para detecção de Clostridium chauvoei em tecidos de Cavia porcellus PCR detection of Clostridium chauvoei in tissues of Cavia porcellus
Embrapa Informação Tecnológica, 2005Co-Authors: Ronnie Antunes De ,assis, Francisco Carlos Faria Lobato, Mariano Fernandez Miyakawa, Francisco Alejandro UzalAbstract:O objetivo deste trabalho foi padronizar uma técnica de reação em cadeia da polimerase (PCR), para detecção de Clostridium chauvoei, em tecidos fixados em formol e incluídos em parafina de cobaias (Cavia porcellus) experimentalmente infectadas com esse microrganismo. Os animais foram sacrificados, e amostras do músculo da área de inoculação (MAI), fígado, miocárdio e baço foram disponibilizadas para a técnica de PCR. O clostrídio foi detectado em todas as secções do MAI, fígado e miocárdio, mas não foi observado em secções do baço. Reações cruzadas não foram observadas a partir de secções do MAI dos animais com inoculação de outras espécies de clostrídios, bem como nenhuma amplificação foi observada a partir de secções do MAI dos animais controle. Esses resultados mostram que a técnica de PCR desenvolvida neste estudo, pode ser usada para detecção de Clostridium chauvoei em tecidos fixados em formol e incluídos em parafina.The objective of this work was the standardization of a polymerase chain reaction (PCR) for detection of Clostridium chauvoei in formalin-fixed, paraffin-embedded tissues of guinea-pigs (Cavia porcellus) infected experimentally with this microorganism. The animals were sacrificed, and samples of muscle from inoculation area (MIA), liver, myocardium and spleen were available for PCR technique. Clostridium chauvoei was detected in all sections of the MIA, liver and myocardium, and no product was observed in sections of the spleen. Cross-reactions were not observed in sections of MIA of the animals inoculated with other clostridia, as well as no amplification was observed in sections of MIA of control animals. These results show that the PCR technique developed in this study may be useful for detection of C. chauvoei in formalin-fixed, paraffin-embedded tissues
Yoshimasa Sasaki - One of the best experts on this subject based on the ideXlab platform.
-
Human Fulminant Gas Gangrene Caused by Clostridium chauvoei
Journal of clinical microbiology, 2008Co-Authors: Noriyuki Nagano, Yoshimasa Sasaki, Shinji Isomine, Haru Kato, Motohide Takahashi, Koji Sakaida, Yukiko Nagano, Yoshichika ArakawaAbstract:The first human case of fulminant gas gangrene caused by Clostridium chauvoei, a pathogen causing ruminant blackleg, was confirmed for a 58-year-old man suffering from diabetes mellitus. The patient developed conspicuous emphysematous gangrene in the right chest wall as well as intravascular gas entrapments and died 2 h after hospital arrival.
-
phylogenetic analysis and pcr detection of Clostridium chauvoei Clostridium haemolyticum Clostridium novyi types a and b and Clostridium septicum based on the flagellin gene
Veterinary Microbiology, 2002Co-Authors: Yoshimasa Sasaki, Noriyasu Takikawa, Akemi Kojima, Hiroshi Aoki, Yasuaki Ogikubo, Yutaka TamuraAbstract:Abstract The flagellin genes ( fliC ) of Clostridium chauvoei , Clostridium haemolyticum , Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene ( fliC ) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I ( Collins et al., 1994 ), are more closely related to Bacillus subtilis than to Clostridium difficile , which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei , C. haemolyticum , C. novyi types A and B, and C. septicum . PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
-
phylogenetic analysis and pcr detection of Clostridium chauvoei Clostridium haemolyticum Clostridium novyi types a and b and Clostridium septicum based on the flagellin gene
Veterinary Microbiology, 2002Co-Authors: Yoshimasa Sasaki, Noriyasu Takikawa, Akemi Kojima, Hiroshi Aoki, Yasuaki Ogikubo, Yutaka TamuraAbstract:The flagellin genes (fliC) of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene (fliC) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I (), are more closely related to Bacillus subtilis than to Clostridium difficile, which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei, C. haemolyticum, C. novyi types A and B, and C. septicum. PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
-
rapid detection and identification of Clostridium chauvoei by pcr based on flagellin gene sequence
Veterinary Microbiology, 2001Co-Authors: Akemi Kojima, Yoshimasa Sasaki, Yasuaki Ogikubo, Ikuo Uchida, Tsutomu Sekizaki, Yutaka TamuraAbstract:We developed a one-step polymerase chain reaction (PCR) system that specifically detects Clostridium chauvoei. Oligonucleotide primers were designed to amplify a 516-bp fragment of the structural flagellin gene. The specificity of the PCR was investigated by analyzing 59 strains of clostridia, and seven strain of other genera. A 516-bp fragment could be amplified from all the C. chauvoei strains tested, and no amplification was observed by using DNAs from the other strains tested, including Clostridium septicum. Similarly, this PCR-based method specifically detected C. chauvoei DNA sequences in samples of muscle and exudate of obtained from mice within 12 h of inoculation. In tests using samples of muscle or liver, the limit of detection was about 200 organisms per reaction. These results suggest that the one-step PCR system may be useful for direct detection and identification of C. chauvoei in clinical specimens.
-
cloning and expression of a gene encoding the flagellin of Clostridium chauvoei
Veterinary Microbiology, 2000Co-Authors: Akemi Kojima, Yoshimasa Sasaki, Yasuaki Ogikubo, Mayumi Kijima, Ikuo Uchida, Tsutomu Sekizaki, Yutaka TamuraAbstract:Abstract Clostridium chauvoei is a causative agent of blackleg and the major protective antigen of the organism is the flagellar protein. Using an Escherichia coli expression library of the C. chauvoei Okinawa strain, we isolated the fliC gene encoding the flagellin protein. DNA sequence analysis revealed an open reading frame of 413 amino acid residues with a calculated molecular mass of 43 819 Da. Comparison of the sequence with those of flagellins from other bacteria showed considerable homology in the N-terminal and C-terminal domains. The glutathione-S-transferase (GST)-flagellin fusion protein and the purified FliC protein after removing the GST part with thrombin reacted with both polyclonal antisera and the non-protective monoclonal antibody (Mab), Mo-114. However, the protective Mab, Mo-41, which may recognize its conformational epitope, failed to react with both the GST-flagellin fusion protein and the purified FliC. Furthermore, the GST-flagellin fusion protein and the purified FliC induced very little protective immunity in mice. These results suggested that a conformation-dependent epitope play an important role in the development of immunity against blackleg.
Yasuaki Ogikubo - One of the best experts on this subject based on the ideXlab platform.
-
phylogenetic analysis and pcr detection of Clostridium chauvoei Clostridium haemolyticum Clostridium novyi types a and b and Clostridium septicum based on the flagellin gene
Veterinary Microbiology, 2002Co-Authors: Yoshimasa Sasaki, Noriyasu Takikawa, Akemi Kojima, Hiroshi Aoki, Yasuaki Ogikubo, Yutaka TamuraAbstract:The flagellin genes (fliC) of Clostridium chauvoei, Clostridium haemolyticum, Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene (fliC) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I (), are more closely related to Bacillus subtilis than to Clostridium difficile, which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei, C. haemolyticum, C. novyi types A and B, and C. septicum. PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
-
phylogenetic analysis and pcr detection of Clostridium chauvoei Clostridium haemolyticum Clostridium novyi types a and b and Clostridium septicum based on the flagellin gene
Veterinary Microbiology, 2002Co-Authors: Yoshimasa Sasaki, Noriyasu Takikawa, Akemi Kojima, Hiroshi Aoki, Yasuaki Ogikubo, Yutaka TamuraAbstract:Abstract The flagellin genes ( fliC ) of Clostridium chauvoei , Clostridium haemolyticum , Clostridium novyi types A and B, and Clostridium septicum were analysed by PCR amplification and DNA sequencing. The five Clostridium species have at least two copies of the flagellin gene ( fliC ) arranged in tandem on the chromosome. The deduced N- and C-terminal aminoacid sequences of the flagellin proteins (FliCs) of these clostridia are well conserved but their central region aminoacid sequences are not. Phylogenic analysis based on the N-terminal aminoacid sequence of the FliC protein revealed that these clostridia, which belong to Clostridium 16S rDNA phylogenic cluster I ( Collins et al., 1994 ), are more closely related to Bacillus subtilis than to Clostridium difficile , which belongs to the cluster XI. Moreover, a multiplex polymerase reaction (PCR) system based on the fliC sequence was developed to rapidly identify C. chauvoei , C. haemolyticum , C. novyi types A and B, and C. septicum . PCR of each Clostridium amplified a species-specific band. The multiplex PCR system may be useful for rapid identification of pathogenic clostridia.
-
amplification of the 16s 23s rdna spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum
Research in Veterinary Science, 2001Co-Authors: Y Sasaki, Yasuaki Ogikubo, Kinya Yamamoto, Mari Norimatsu, K Amimoto, A Kojima, H Ogata, Yutaka TamuraAbstract:Amplification of the 16S-23S rDNA spacer region by polymerase chain reaction (PCR) was used for the rapid detection of Clostridium chauvoei and C septicum. To assess its specificity, PCR was performed with total DNA from 42 strains of clostridia and three strains of other genera. PCR products specific to C chauvoei or to C septicum were generated from homologous cultures only. Clostridium chauvoer-specific or C septicum-specific amplicons were also generated from tissues of cows experimentally infected with C chauvoei or C septicum and in DNA samples from cows clinically diagnosed as having blackleg or malignant oedema. These results suggest that a species-specific PCR may be useful for the rapid and direct detection of C chauvoei and C septicum in clinical specimens.
-
amplification of the 16s 23s r dna spacer region for rapid detection of Clostridium chauvoei and Clostridium septicum
Research in Veterinary Science, 2001Co-Authors: Y Sasaki, Yasuaki Ogikubo, Kinya Yamamoto, Mari Norimatsu, K Amimoto, A Kojima, H Ogata, Yutaka TamuraAbstract:Abstract Amplification of the 16S-23S r DNA spacer region by polymerase chain reaction ( PCR ) was used for the rapid detection of Clostridium chauvoei and C septicum . To assess its specificity, PCR was performed with total DNA from 42 strains of clostridia and three strains of other genera. PCR products specific to C chauvoei or to C septicum were generated from homologous cultures only. Clostridium chauvoer -specific or C septicum -specific amplicons were also generated from tissues of cows experimentally infected with C chauvoei or C septicum and in DNA samples from cows clinically diagnosed as having blackleg or malignant oedema. These results suggest that a species-specific PCR may be useful for the rapid and direct detection of C chauvoei and C septicum in clinical specimens.
-
rapid detection and identification of Clostridium chauvoei by pcr based on flagellin gene sequence
Veterinary Microbiology, 2001Co-Authors: Akemi Kojima, Yoshimasa Sasaki, Yasuaki Ogikubo, Ikuo Uchida, Tsutomu Sekizaki, Yutaka TamuraAbstract:We developed a one-step polymerase chain reaction (PCR) system that specifically detects Clostridium chauvoei. Oligonucleotide primers were designed to amplify a 516-bp fragment of the structural flagellin gene. The specificity of the PCR was investigated by analyzing 59 strains of clostridia, and seven strain of other genera. A 516-bp fragment could be amplified from all the C. chauvoei strains tested, and no amplification was observed by using DNAs from the other strains tested, including Clostridium septicum. Similarly, this PCR-based method specifically detected C. chauvoei DNA sequences in samples of muscle and exudate of obtained from mice within 12 h of inoculation. In tests using samples of muscle or liver, the limit of detection was about 200 organisms per reaction. These results suggest that the one-step PCR system may be useful for direct detection and identification of C. chauvoei in clinical specimens.