AIM:To examine the detection rate of viable Mycobacterium avium subspecies paratuberculosis(MAP) in patients with inflammatory bowel disease [Crohn's disease(CD) and ulcerative colitis(UC)].METHODS:Thirty patients...AIM:To examine the detection rate of viable Mycobacterium avium subspecies paratuberculosis(MAP) in patients with inflammatory bowel disease [Crohn's disease(CD) and ulcerative colitis(UC)].METHODS:Thirty patients with CD(15 with at least one NOD2/CARD15 mutation),29 with UC,and 10 with no inflammatory bowel disease(IBD).were tested for MAP by polymerase chain reaction(specific IS900 fragment) and blood culture.RESULTS:MAP DNA was detected in all original blood samples and 8-wk blood cultures(CD,UC and non-IBD).Positive MAP DNA status was confirmed by dot blot assays.All 69 cultures were negative by acid-fast Ziehl-Neelsen staining.Viable MAP,in spheroplast form,was isolated from the 18-mo blood cultures of all 30 CD patients,one UC patient,and none of the non-IBD controls.No association was found between positive MAP cultures and use of immunosuppressive drugs or CDassociated single nucleotide polymorphisms.CONCLUSION:MAP is widely present in our area and MAP DNA can be recovered from the blood of CD,UC and non-IBD patients.However,MAP spheroplasts were only found in CD patients.展开更多
Mycobacterium avium subspecies paratuberculosis(MAP),originally called Johne’s bacillus was firstdescribed from Germany in 1895 as the cause of achronic inflammatory disease of the intestine in acow.As the 20th centu...Mycobacterium avium subspecies paratuberculosis(MAP),originally called Johne’s bacillus was firstdescribed from Germany in 1895 as the cause of achronic inflammatory disease of the intestine in acow.As the 20th century progressed,clinical andsub-clinical MAP infection in farm animals inWestern Europe appeared to become moreprevalent.Among the early reviews available展开更多
The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne's dis...The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne's disease is a granulomatous chronic enteritis of cattle and sheep caused by Mycobacterium avium subspecies paratuberculosis(MAP) and shows some analogies with Crohn's disease(CD). Several studies have tried to clarify if MAP has a role in the etiology of CD. The present article provides an overview of the evidence in favor and against the "MAP-hypothesis", analyzing the methods commonly adopted to detect MAP and the role of antimycobacterial therapy in patients with inflammatory bowel disease. Studies were identified through the electronic database, MEDLINE, and were selected based on their relevance to the objective of the review. The presence of MAP was investigated using multiple diagnostic methods for MAP detection and in different tissue samples from patients affected by CD or ulcerative colitis and in healthy controls. On the basis of their studies, several authors support a close relationship between MAP and CD. Although increasing evidence of MAP detection in CD patients is unquestionable, a clear etiological link still needs to be proven.展开更多
This study characterizes the 19 kDa protein expressed by Mycobacterium avium subspecies paratuberculosis (MAP) as a glycolipoprotein, providing the foundation for future experiments regarding its antigenicity and role...This study characterizes the 19 kDa protein expressed by Mycobacterium avium subspecies paratuberculosis (MAP) as a glycolipoprotein, providing the foundation for future experiments regarding its antigenicity and role in disease pathogenicity. We have previously shown that a 4.8 kb insert from MAP will produce a 16 kDa recombinant protein when expressed in Escherichia coli and 19 kDa recombinant protein when expressed in M. smegmatis (smeg19K). The difference of 3 kDa in size of these expressed proteins may be related to post translational modifications that occur in Mycobacterium species. We hypothesized that smeg19K is a glycolipoprotein since BLAST analysis revealed approximately 76% amino acid identity between the MAP 19 kDa protein and a known lipoglycoprotein, the 19 kDa protein of M. tuberculosis. This prediction was confirmed by the following positive staining of smeg19K with Sudan Black 4B, a postelectrophoresis dye used to stain for lipids. Smeg19K has also stained positively for glycosylation with the lectin concavalin A, a highly specific stain for mannose residues. As expected, treatment with tunicamycin (an antibiotic known to inhibit N-glycosylation) and treatment with deglycosylation assay (non-specific for mannose), showed no reduction in size of 19 kDa glycolipoprotein.展开更多
Objective:To isolate and identify the exact species of the genus Mycobacterium from Didelphis(D.)virginiana,and the direct implications of this bacterium to public health and veterinary medicine.Methods:Thirty-one D.v...Objective:To isolate and identify the exact species of the genus Mycobacterium from Didelphis(D.)virginiana,and the direct implications of this bacterium to public health and veterinary medicine.Methods:Thirty-one D.virginiana were captured and necropsied in Hidalgo,Mexico.Tissue samples were collected to culture mycobacteria present and examine individual specimens’histopathology.Mycobacterium identification was obtained through the application of amplification and sequencing of 16S rDNA techniques.Results:Three strains were isolated and identified as Mycobacterium(M.)avium subsp.hominissuis by utilizing M.avium complexspecific primers.Granulomatous lesions were observed in the subpleural zone(granuloma gradeⅡ)and bronchial(granuloma gradeⅠ)of the lungs of D.virginiana with positive isolation.Conclusions:Three strains of M.avium subsp.hominissuis,from lung tissue samples of D.virginiana were identified.This subspecies of M.avium has important implications in public health and veterinary medicine.展开更多
Johne's disease or paratuberculosis is a chronic mycobacterial infection that affects cattle, sheep, goats and other ruminants, adversely, leading to huge economic losses throughout the world. The estimation of se...Johne's disease or paratuberculosis is a chronic mycobacterial infection that affects cattle, sheep, goats and other ruminants, adversely, leading to huge economic losses throughout the world. The estimation of sero-prevalence of this disease in the cattle population of south-western Bangalore, Karnataka, using an immunological assay and statistical analyses, was the objective of this study. One of the diagnostic tools used to detect an antigen or an antibody in animal serum or milk is the Enzyme Linked Immuno-Sorbent Assay, which has been widely used in the research and diagnosis of animal and human diseases as its accuracy is of nanogram-picogram/milliltre level. In the present study, indirect-ELISA was used to diagnose and estimate the sero-prevalence of paratuberculosis in cattle showing diarrhoea and/or anaemia, at 5 local dairy farms in south-west Bangalore, India. Out of 350 bovine serum samples, 53 (15.14%) were positive, 55 milk samples out of 300 were found positive (18.33%) for antibody against Johne's disease by indirect ELISA. The positive samples were then confirmed by direct smear examination of dung by Ziehl-Neelsen staining. Statistical analyses were carried out to indicate the seroprevalence of Johne's disease in the cattle population of this region to be 15 ± 10%, taking a confidence interval of 95%. The results emphasize the need to prevent the further spread of infection to other susceptible animals and humans as the causative organism, Mycobacterium avium subsp. paratuberculosis is implicated in Crohn's disease, an irritable bowel syndrome in humans.展开更多
基金Supported by A grant from Fundación de Investigación Médica Mutua Madrilena(FMM)(to Díaz-Rubio M)
文摘AIM:To examine the detection rate of viable Mycobacterium avium subspecies paratuberculosis(MAP) in patients with inflammatory bowel disease [Crohn's disease(CD) and ulcerative colitis(UC)].METHODS:Thirty patients with CD(15 with at least one NOD2/CARD15 mutation),29 with UC,and 10 with no inflammatory bowel disease(IBD).were tested for MAP by polymerase chain reaction(specific IS900 fragment) and blood culture.RESULTS:MAP DNA was detected in all original blood samples and 8-wk blood cultures(CD,UC and non-IBD).Positive MAP DNA status was confirmed by dot blot assays.All 69 cultures were negative by acid-fast Ziehl-Neelsen staining.Viable MAP,in spheroplast form,was isolated from the 18-mo blood cultures of all 30 CD patients,one UC patient,and none of the non-IBD controls.No association was found between positive MAP cultures and use of immunosuppressive drugs or CDassociated single nucleotide polymorphisms.CONCLUSION:MAP is widely present in our area and MAP DNA can be recovered from the blood of CD,UC and non-IBD patients.However,MAP spheroplasts were only found in CD patients.
文摘Mycobacterium avium subspecies paratuberculosis(MAP),originally called Johne’s bacillus was firstdescribed from Germany in 1895 as the cause of achronic inflammatory disease of the intestine in acow.As the 20th century progressed,clinical andsub-clinical MAP infection in farm animals inWestern Europe appeared to become moreprevalent.Among the early reviews available
文摘The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne's disease is a granulomatous chronic enteritis of cattle and sheep caused by Mycobacterium avium subspecies paratuberculosis(MAP) and shows some analogies with Crohn's disease(CD). Several studies have tried to clarify if MAP has a role in the etiology of CD. The present article provides an overview of the evidence in favor and against the "MAP-hypothesis", analyzing the methods commonly adopted to detect MAP and the role of antimycobacterial therapy in patients with inflammatory bowel disease. Studies were identified through the electronic database, MEDLINE, and were selected based on their relevance to the objective of the review. The presence of MAP was investigated using multiple diagnostic methods for MAP detection and in different tissue samples from patients affected by CD or ulcerative colitis and in healthy controls. On the basis of their studies, several authors support a close relationship between MAP and CD. Although increasing evidence of MAP detection in CD patients is unquestionable, a clear etiological link still needs to be proven.
文摘This study characterizes the 19 kDa protein expressed by Mycobacterium avium subspecies paratuberculosis (MAP) as a glycolipoprotein, providing the foundation for future experiments regarding its antigenicity and role in disease pathogenicity. We have previously shown that a 4.8 kb insert from MAP will produce a 16 kDa recombinant protein when expressed in Escherichia coli and 19 kDa recombinant protein when expressed in M. smegmatis (smeg19K). The difference of 3 kDa in size of these expressed proteins may be related to post translational modifications that occur in Mycobacterium species. We hypothesized that smeg19K is a glycolipoprotein since BLAST analysis revealed approximately 76% amino acid identity between the MAP 19 kDa protein and a known lipoglycoprotein, the 19 kDa protein of M. tuberculosis. This prediction was confirmed by the following positive staining of smeg19K with Sudan Black 4B, a postelectrophoresis dye used to stain for lipids. Smeg19K has also stained positively for glycosylation with the lectin concavalin A, a highly specific stain for mannose residues. As expected, treatment with tunicamycin (an antibiotic known to inhibit N-glycosylation) and treatment with deglycosylation assay (non-specific for mannose), showed no reduction in size of 19 kDa glycolipoprotein.
基金the financial assistance from the Secretary of Research of Universidad Autónoma del Estado de Hidalgo(UAEH)through the project“Epidemiological surveillance of the genus Mycobacterium in the species Didelphis virginiana(Tlacuache)of the valley of Tulancingo Hidalgo”with number UAEH/DI/ICAp/MVZ/6.
文摘Objective:To isolate and identify the exact species of the genus Mycobacterium from Didelphis(D.)virginiana,and the direct implications of this bacterium to public health and veterinary medicine.Methods:Thirty-one D.virginiana were captured and necropsied in Hidalgo,Mexico.Tissue samples were collected to culture mycobacteria present and examine individual specimens’histopathology.Mycobacterium identification was obtained through the application of amplification and sequencing of 16S rDNA techniques.Results:Three strains were isolated and identified as Mycobacterium(M.)avium subsp.hominissuis by utilizing M.avium complexspecific primers.Granulomatous lesions were observed in the subpleural zone(granuloma gradeⅡ)and bronchial(granuloma gradeⅠ)of the lungs of D.virginiana with positive isolation.Conclusions:Three strains of M.avium subsp.hominissuis,from lung tissue samples of D.virginiana were identified.This subspecies of M.avium has important implications in public health and veterinary medicine.
文摘Johne's disease or paratuberculosis is a chronic mycobacterial infection that affects cattle, sheep, goats and other ruminants, adversely, leading to huge economic losses throughout the world. The estimation of sero-prevalence of this disease in the cattle population of south-western Bangalore, Karnataka, using an immunological assay and statistical analyses, was the objective of this study. One of the diagnostic tools used to detect an antigen or an antibody in animal serum or milk is the Enzyme Linked Immuno-Sorbent Assay, which has been widely used in the research and diagnosis of animal and human diseases as its accuracy is of nanogram-picogram/milliltre level. In the present study, indirect-ELISA was used to diagnose and estimate the sero-prevalence of paratuberculosis in cattle showing diarrhoea and/or anaemia, at 5 local dairy farms in south-west Bangalore, India. Out of 350 bovine serum samples, 53 (15.14%) were positive, 55 milk samples out of 300 were found positive (18.33%) for antibody against Johne's disease by indirect ELISA. The positive samples were then confirmed by direct smear examination of dung by Ziehl-Neelsen staining. Statistical analyses were carried out to indicate the seroprevalence of Johne's disease in the cattle population of this region to be 15 ± 10%, taking a confidence interval of 95%. The results emphasize the need to prevent the further spread of infection to other susceptible animals and humans as the causative organism, Mycobacterium avium subsp. paratuberculosis is implicated in Crohn's disease, an irritable bowel syndrome in humans.
