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PubMed2006

Sequencing of hsp65 distinguishes among subsets of the Mycobacterium avium complex.

The Mycobacterium avium complex consists of epidemiologically distinct subsets. The classification of these subsets is complicated by a number of factors, including the ambiguous results obtained with phenotypic and genetic assays and the recent appreciation that human and avian strains appear to be distinct. In previous work, sequencing based on a 441-bp portion of the hsp65 gene has proven to efficiently classify isolates within the Mycobacterium genus but provides low resolution for distinguishing among members of the M. avium complex. Therefore, in this study, we have targeted the more variable 3' region of the hsp65 gene to determine whether it can effectively discriminate M. avium complex isolates at the levels of species and subspecies. Primers designed for this target consistently generated amplicons for all organisms classified as M. avium complex. Sequences obtained indicate that M. intracellulare is genetically divergent from M. avium organisms, and distinct sequevars were obtained for M. avium subsets, including M. avium subsp. avium (bird type), M. avium subsp. hominissuis, and M. avium subsp. paratuberculosis. In addition, sequence differences served to distinguish bovine from ovine strains of M. avium subsp. paratuberculosis. A unique profile for M. avium subsp. silvaticum was not obtained. These results indicate that sequencing the 3' region of the hsp65 gene can simply and unambiguously distinguish species and subspecies of the M. avium complex.

Animals↗
PubMed1980

Agglutinins to bacteria in Crohn's disease.

Sera from patients with Crohn's disease were tested for antibodies against organisms which are thought to cause inflammatory bowel disease in animals, or have been implicated in human Crohn's disease. Control sera were collected from healthy individuals and patients with ulcerative colitis. Sera from Crohn's disease and controls failed to agglutinate Clostridium colinum or Campylobacter sputorum subsp. mucosalis and two strains of Mycobacterium paratuberculosis (M26 and M27). Most of the sera agglutinated a Citrobacter freundii variant, Mycobacterium paratuberculosis (M28) and Mycobacterium avium (M41) but Crohn's disease sera did not differ from controls. A complement fixation test against Chlamydia gave more positive reactions in patients with Crohn's disease and colitis than in healthy controls. There was a clear difference between the sera from patients with Crohn's disease and other sera, including ulcerative colitis, in agglutination tests with the commensal coccoid rods of the genera Eubacterium and Peptostreptococcus; in these tests 54% of sera from Crohn's disease were positive compared with 11% in ulcerative colitis and none of the sera from healthy controls. All the results were essentially negative with the exception of those from Eubacterium and Peptostreptococcus and these bacteria merit investigation.

Agglutination Tests↗
PubMed2000

Sensitive differential detection of genetically related mycobacterial pathogens in archival material.

A polymerase chain reaction (PCR) assay targeted to the immunogenic protein MPB64 gene was used to detect members of the Mycobacterium tuberculosis complex, and an outward-primed PCR (OPPCR) designed on the IS6110 element allowed differentiation between Mycobacterium bovis and Mycobacterium tuberculosis. Additionally, the amplification of IS1110 and 16S ribosomal RNA sequences combined with a dot blotting assay were able to differentially detect Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium paratuberculosis. The validity of the experimental procedure was tested on reference material and formalin-fixed paraffin-embedded samples from patients with tuberculosis, sarcoidosis, or Crohn disease. We demonstrated mycobacterial DNA in 59 of 75 cases with histologic lesions typical of tuberculosis; we detected M tuberculosis and M paratuberculosis in 6 of 25 sarcoidosis cases and in 7 of 20 Crohn disease specimens, respectively. The proposed diagnostic procedure is directly applicable to archival material and allows differentiation of genetically related mycobacterial pathogens in more detail than other molecular methods. It provides a tool for the diagnostic study of tuberculosis, sarcoidosis, and Crohn disease.

Antigens, Bacterial↗
PubMed1999

A case of intestinal Mycobacterium simiae infection in an SIV-infected immunosuppressed rhesus monkey.

Although Mycobacterium simiae was identified and classified more than three decades ago, only a few cases are mentioned in the current literature. After experimental simian immunodeficiency virus infection, a 9-year-old female rhesus monkey (Macaca mulatta) developed progressive immunosuppression and gastrointestinal disease very similar to the clinical and pathomorphologic features of Johne's disease, which is caused by M. paratuberculosis. Acid-fast-positive bacteria reacted immunohistochemically with antibodies against M. paratuberculosis and M. bovis but were not useful for differentiation because of a high degree of cross-reactivity. In contrast to immunohistochemistry and histopathology, biochemical methods and cycle sequencing analysis of the 16S ribosomal RNA identified M. simiae as the disease-causing pathogen. This case demonstrates the importance of molecular biological methods for the diagnosis of M. simiae infection in monkeys.

Animals↗
PubMed2001

Mycobacterium avium and Mycobacterium intracellulare infection in mammals.

Mycobacterium avium subsp. avium and M. intracellulare are ubiquitous organisms in the environment. The reservoir of M. avium subsp. avium is generally accepted to be environmental, in particular, water and soil are sources of the organism. In contrast to M. avium infection in wild and domestic birds, M. avium infection in mammals occurs only sporadically and is rarely transmissible. Generalised disease is usually uncommon, owing to the non-progressive, chronic character of the infection. However, some cases of disseminated disease have been reported, e.g. in captive non-domestic hoofed animals as well as in immunosuppressed dogs and cats. The majority of M. avium and M. intracellulare infections in livestock are detected at slaughter and the diagnosis is confirmed by bacteriological procedures. Condemnation of affected portions of the carcass can result in significant economic losses, although gross lesions are mostly restricted to lymph nodes close to the alimentary tract. Successful treatment with antibiotics in combination with surgery has been reported in some affected domestic cats, but is not considered to be effective or economical in other species. In the past, differentiation of M. avium bacteria from the closely related M. avium subsp. paratuberculosis was based on the mycobactin dependence and prolonged incubation period of the latter. More recently, amplification of the genomic insertion sequence IS900 has proved to be a powerful tool for identification of M. avium subsp. paratuberculosis. The potential zoonotic importance of M. avium infections has been indicated, but requires clarification.

Animals↗
PubMed2005

Protothecal enteritis as a cause of protein-losing enteropathy in a bull.

Prototheca spp are achlorophyllic saprophytic algae found in wastewater, sewage, agricultural waste, and possibly elsewhere in the environment. Infections with these organisms have been reported in cattle, humans, and dogs; affected cattle commonly develop mastitis. A 5-year-old Brahman-cross bull was evaluated because of a history of diarrhea and weight loss. The history and physical examination and clinicopathologic findings were similar to those associated with granulomatous enteritis caused by Mycobacterium avium subsp paratuberculosis (Johne's disease), which is the most common protein-losing enteropathy of cattle. However, diagnostic tests for paratuberculosis yielded negative results. Biopsy specimens from the ileum, jejunum, and ileocecal lymph node were collected for histologic examination and preparation of tissue impression smears; Prototheca-like organisms were identified. Because of the poor prognosis associated with this infection and the lack of safe and economical therapeutic agents for cattle, the owner decided to euthanatize the bull. Infection with Prototheca organisms was confirmed postmortem. As this case illustrates, protothecosis may be a cause of granulomatous enteritis in cattle.

Animals↗
PubMed1997

Detection of Mycobacterium avium complex in cerebrospinal fluid of a sarcoid patient by specific polymerase chain reaction assays.

The etiology of sarcoidosis is unknown, but it has long been suspected to be mycobacterial. In the present study, we used 4 mycobacterial species-specific polymerase chain reaction assays on cerebrospinal fluid obtained from a patient with neurosarcoidosis. Positive hybridization was observed with both the Mycobacterium avium complex probe and the insertion element IS900-specific probe that has been found in M. paratuberculosis species. There was no hybridization with M. tuberculosis or M. avium woodpigeon strain-specific probes. This case report demonstrates that M. paratuberculosis or some closely related M. avium spp which perhaps also carry IS900, or contain closely related DNA sequences, are associated with at least some cases of sarcoidosis disease.

Adult↗
PubMed2003

Microbiological quality of raw goat's and ewe's bulk-tank milk in Switzerland.

A total of 407 samples of bulk-tank milk (344 of goat's milk and 63 of ewe's milk) collected from 403 different farms throughout Switzerland, was examined. The number of farms investigated in this study represents 8% of the country's dairy-goat and 15% of its dairy-sheep farms. Standard plate counts and Enterobacteriaceae counts were performed on each sample. Furthermore, the prevalence of Staphylococcus aureus, Campylobacter spp., Shiga toxin-producing Escherichia coli, Salmonella spp., and Mycobacterium avium ssp. paratuberculosis was studied. The median standard plate count for bulk-tank milk from small ruminants was 4.70 log cfu/ml (4.69 log cfu/ml for goat's milk and 4.78 log cfu/ml for ewe's milk), with a minimum of 2.00 log cfu/ml and a maximum of 8.64 log cfu/ml. Enterobacteriaceae were detected in 212 (61.6%) goat's milk and 45 (71.4%) ewe's milk samples, whereas S. aureus was detected in 109 (31.7%) samples of goat's milk and 21 (33.3%) samples of ewe's milk. Campylobacter spp. and Salmonella spp. were not isolated from any of the samples. However, 16.3% of the goat's milk and 12.7% of the ewe's milk samples were polymerase chain reaction (PCR)-positive for Shiga toxin-producing E. coli. Seventy-nine (23.0%) goat's tank-milk and 15 (23.8%) ewe's tank-milk samples were PCR-positive for insertion sequence 900, providing presumptive evidence for the presence of M. avium ssp. paratuberculosis. These results form the basis for determining the microbiological quality standards for goat's and ewe's milk. Moreover, the data presented form part of the risk assessment program for raw milk from small ruminants in Switzerland.

Animals↗
PubMed2005

Improving food safety within the dairy chain: an application of conjoint analysis.

This study determined the relative importance of attributes of food safety improvement in the production chain of fluid pasteurized milk. The chain was divided into 4 blocks: "feed" (compound feed production and its transport), "farm" (dairy farm), "dairy processing" (transport and processing of raw milk, delivery of pasteurized milk), and "consumer" (retailer/catering establishment and pasteurized milk consumption). The concept of food safety improvement focused on 2 main groups of hazards: chemical (antibiotics and dioxin) and microbiological (Salmonella, Escherichia coli, Mycobacterium paratuberculosis, and Staphylococcus aureus). Adaptive conjoint analysis was used to investigate food safety experts' perceptions of the attributes' importance. Preference data from individual experts (n = 24) on 101 attributes along the chain were collected in a computer-interactive mode. Experts perceived the attributes from the "feed" and "farm" blocks as being more vital for controlling the chemical hazards; whereas the attributes from the "farm" and "dairy processing" were considered more vital for controlling the microbiological hazards. For the chemical hazards, "identification of treated cows" and "quality assurance system of compound feed manufacturers" were considered the most important attributes. For the microbiological hazards, these were "manure supply source" and "action in salmonellosis and M. paratuberculosis cases". The rather high importance of attributes relating to quality assurance and traceability systems of the chain participants indicates that participants look for food safety assurance from the preceding participants. This information has substantial decision-making implications for private businesses along the chain and for the government regarding the food safety improvement of fluid pasteurized milk.

Animal Feed↗
PubMed2000

[Mycobacterium avium-intracellulare complex: phenotypic and genotypic markers and the molecular basis for interspecies transmission].

The Mycobacterium avium complex (MAC) comprises a heterogeneous group of slowly-growing mycobacteria that are pathogenic for both humans and animals. Two genetically distinct species within MAC are M. avium, which tends to infect HIV-infected patients, and M. intracellulare more common among immunocompetent individuals. Contrary to M. intracellulare which relates to a single species, M. avium is separated into three subspecies; M. avium subsp. avium, a major opportunistic pathogen leading to a disseminated disease among terminal AIDS patients; M. avium subsp. paratuberculosis, causing Johne's disease among ruminants and implicated in Crohn's disease among humans; and M. avium subsp. silvaticum, a pathogen affecting birds that may cause chronic enteritis among calves but has not yet been associated with human disease. With the exception of mycobactin-dependent growth of M. paratuberculosis, most of the biochemical and cultural tests cannot discriminate among the three subspecies of M. avium. However, recently developed molecular methods and fingerprinting of strains using insertion sequences allows not only to distinguish among them but also further to explore the polymorphism of human and animal isolates. Numerous studies have underlined the probable role of various ecological niches (water, dust, soil, pigs, poultry and ruminants etc.) as a possible source of contamination for AIDS patients. This paper reviews the phenotypic and genotypic markers and epidemiology of M. avium complex organisms and current knowledge of the molecular basis of of inter-species transmission.

Animals↗
PubMed1992

Distinctive western blot antibody patterns induced by infection of mice with individual strains of the Mycobacterium avium complex.

Systemic infection of mice with organisms of the Mycobacterium avium complex (MAC) induced antibody responses, characteristic for each of the three tested individual strains. The influence of host genetic factors was reflected up to 3 months after infection by the finding of generally oligobanded and multibanded Western blot patterns in C57B1/6 and BALB/c mice, respectively. Nevertheless, more bands developed at 6 months in C57BL/6 mice. The response to three antigens of 18,000, 38,000 and 24,000 MW was analysed in greater detail. Antibodies to a protease-resistant 18,000 MW band produced only by BALB/c mice were either strain specific, following infection with M. avium, strain Maa-B2, or cross-reactive within MAC, following infection with M. avium strain Maa-A6 and M. paratuberculosis, strain Map-203. Another protease-resistant antigen of 38,000 MW was immunogenic only in Maa-B2 infected mice. This constituent was found to be related to the protease-sensitive antigen of corresponding molecular weight from M. tuberculosis. Two 24,000 MW proteins of M. paratuberculosis were separated by two-dimensional gel electrophoresis: antibodies to the anodic band were induced by Map-203 infection, whilst the cathodic band was revealed by heteroclitic antibodies from Maa-B2-infected mice. The latter antigen is apparently expressed during in vivo replication, but not during in vitro culture of Maa-B2 bacteria. We generally conclude, that the selective antibody patterns after live infection, could be attributed to differences in the release of native antigens within mycobacterial lesions. In view of a high degree of species specificity, some of the immunogenic constituents identified may also be useful for serodiagnostic application.

Animals↗
PubMed1990

Studies on heterocyclic compounds: spiro [indole-3,2'-thiazolidine] derivatives. Antimicrobial activity of monohalogenated 3'-phenylspiro 3H-indole-3,2'-thiazolidine-2,4' (1H)-diones.

The following halogenated 3'-phenyl [3H-indole-3,2'-thiazolidine]-2,4'(1H)-dione of general formula (A) were synthesized and screened for antimicrobial activity. (formula: see text) where: X = H (I, III, V, VII, IX, XI, XIII, XV), CH3 (II, IV, VI, VIII, X, XII, XIV, XVI); Y = H (I, II), 3-F (III, IV), 2-Cl (V, VI), 3-Cl (VII, VIII), 4-Cl (IX, X), 2-Br (XI, XII), 3-Br (XIII, XIV), 4-Br (XV, XVI). The synthetic approach involves the preparation of variously substituted Schiff-bases of indol-2,3-dione, which then are subjected to cyclocondensation with alpha-mercaptoalkanoic acids, to give spirothiazolidinones of type (A). The prepared compounds were screened against S. aureus, B. cereus, M. paratuberculosis, E. coli, S. typhi, Pr. mirabilis, Ps. aeruginosa, C. albicans, S. cerevisiae, A. niger by a disk-diffusion assay (Kirby-Bauer modified. The results of the antimicrobial screening showed that the prepared compounds exhibited varying degrees of activity against Gram-positive, Gram-negative bacteria, and fungi. 3-Fluoro-derivative (III) showed inhibitory activity especially toward S. aureus and C. albicans. Chloroderivatives (VII) and (VIII) showed broad-spectrum "in vitro" antimicrobial activity, and were especially inhibitory toward S. aureus, E. coli, and S. Typhi. Fluoro-derivative (IV) and bromo-derivatives (XIII) and (XIV) possessed marked antimicrobial activity against M. paratuberculosis.

Anti-Bacterial Agents↗
PubMed1990

Studies on heterocyclic compounds: spiro [indole-3,2'-thiazolidine] derivatives. II. Antimicrobial activity of halogenated 3'-phenylspiro 3H-indole-3,2'-thiazolidine -2,4 (1H)-diones.

The following polyhalogenated 3'-phenyl 3H-indole-3,2'-thiazolidine -2,4' (1H)-dione of general formula (A) were synthesized and screened for antimicrobial activity. (formula: see text) where: X = H (I, III, V, VII, IX, XI), CH3 (II, IV, VI, VIII, X, XII); Y = H (I, II), 2,4-F2 (III, IV), 2,4-Cl2 (V, VI), 3,4-Cl2 (VII, VIII), 2,6-Cl2 (IX, X), 2,4,6-Cl3 (XI, XII). The general synthetic route involves the preparation of variously substituted isatin-3-imines, which are subjected to cyclocondensation with thioglycolic acid to give compounds I, III, V, VII, IX, XI, or thiolactic acid to give compounds II, IV, VI, VII, X, XII. The prepared compounds were screened against S. aureus, B. cereus, M. paratuberculosis, E. coli, Pr. mirabilis, Ps. aeruginosa, C. albicans, S. cerevisiae, A. niger by a disk-diffusion assay (Kirby-Bauer modified). The results of the antimicrobial screening showed that the polyhalogenated derivatives of type (A) exhibited varying degrees of activity against Gram-positive, Gram-negative bacteria, and fungi. Compound (III) showed a significant activity toward A. niger, moreover compound (IV) was active toward C. albicans. Compound (IX) was very active toward S. typhi and Ps. aeruginosa. Compounds (VII), (IX) and (XII) were very active toward M. paratuberculosis.

Anti-Bacterial Agents↗
PubMedDate not supplied

Absence of mycobactin in Mycobacterium leprae; probably a microbe dependent microorganism implications.

Ferric mycobactins were prepared from Mycobacterium phlei. Mycobacterium avium--intracellulare A and H, isolated respectively from armadillo and human leprosy specimens. Attempts were made to extract mycobactin from host grown M. leprae cells. The crude ferric mycobactin extracts were tested for growth supporting effect on the mycobactin dependent M. paratuberculosis strain ATCC 19698. Mycobactins prepared from M. phlei and the two M. avium--intracellulare strains had growth promoting effect on M. paratuberculosis. The same test organism did not grow in media supplemented with the extract prepared from M. leprae. Results indicate the absence of mycobactin from host grown M. leprae. Since M. leprae cells contain cytochrome c and since mycobactin is essential to growth of all mycobacteria, M. leprae might be considered as a microbe dependent microbe. It is proposed that secondary mycobacteria present in M. leprae infected humans and armadillos might provide mycobactin for in vivo multiplication of M. leprae.

Iron↗
PubMed1984

Biological activity of 4-hydroxyisophthalic acid derivatives. II. Anilides with antimicrobial activity.

A series of 1,3 -bis-anilides of 4-hydroxyisophthalic acid was prepared and tested for antibacterial and antifungal activity. The prepared compounds (I-XVIII), of general structure (A), (Formula: see text) where Xn = H (I); 2-F (II); 3-F (III); 4-F (IV); 2-Cl (V); 3-Cl (VI); 4-Cl (VII); 2-Br (VIII); 3-Br (IX); 4-Br (X); 2-J (XI); 3-J (XII); 4-J (XIII); 2,5-Cl2 (XIV); 2,4-Br2 (XV); 2,3,4-Cl3 (XVI), 2,4,5-Cl3 (XVII); 2,4,6-Cl3 (XVIII), were investigated for the purpose of determining the effect of halogen-substitution on the aniline rings of (A). All of these compounds were prepared in satisfactory hield by reaction of 4-hydroxyisophthalic acid with the appropriate aromatic amine at 175 degrees for 3 hours. The 1,3-bis-anilides prepared in this investigation were screened for antimicrobial activity by a disk-diffusion assay (Kirby-Bauer modified). The organisms used were laboratory cultures of S. aureus, B. subtilis, B. anthracis, M. paratuberculosis 607, E. coli Bb, S. typhi, S. typhimurium, S. paratyphi B, Pr. vulgaris, Kl. pneumoniae, Ps. aeruginosa, C. albicans, and A. niger. The results of this investigation indicated that most of the 1,3-bis-(halogen-anilides) of 4-hydroxyisophthalic acid had little or no antifungal activity "in vitro", while showed significant activity against Gram+ and Gram- bacteria. Some fluoro-derivatives showed inhibitory activity especially toward S. aureus and M. paratuberculosis. Iodo-derivatives showed broad-spectrum "in vitro" antimicrobial activity, and had some antifungal activity.

Anilides↗
PubMed1983

Biological activity of 4-hydroxy-5-formylbenzoic acid derivatives.

A group of thirty 4-hydroxy-5-formylbenzoic acid derivatives of general formula: (formula; see text) where (formula; see text) have been prepared and characterized in an attempt to make available for testing a representative selection of hitherto undescribed hydroxyformylbenzoic acid derivatives. The products, which are listed in Table I, with pertinent data, have been obtained in satisfactory yield and in a good state of purity. The prepared compounds have been tested for "in vitro" activity against three fungi (A. niger, C. albicans, S. cerevisiae), six Gram- bacteria (E. coli Bb, S. typhi, S. infantis, S. paratyphi A, B. melitensis, P. mirabilis) and four Gram+ bacteria (M. paratuberculosis ATCC 607; S. epidermidis, B. subtilis, B. anthracis), by the agar diffusion method (Kirby-Bauer modified). In general, the results of the tests indicated that most of the compounds: a) didn't exhibit antifungal activity "in vitro"; b) had little activity on Gram- bacteria; showed inhibitory activity toward Gram+ species, in particular toward M. paratuberculosis. Other activities of pharmaceutical interest will be tested; the screening program includes tests for cardiac activity. The results of these studies will be published as soon as they are complete.

Bacteria↗
PubMed1982

[Hydrazide derivatives of fluorobenzoic acids and their antimicrobial activity].

The compounds of formula (A) were prepared by reaction between Rn-substituted benzoyl hydrazides and R-substituted benzoyl chlorides; the compounds of formula (B) were obtained by condensing R-substituted benzoyl chlorides with isonicotinic hydrazide. All the compounds were tested for in vitro activity against five Gram+ bacteria (S. aureus, S. epidermidis, B. subtilis, B. anthracis, M. paratuberculosis ATCC 607), seven Gram-bacteria (S. paratyphi A, S. paratyphi B, E. coli, B. abortus, B. melitensis, P. aeruginosa, P. mirabilis) and three fungi (C. albicans, A. niger, S. cerevisiae), by agar-diffusion method (Kirby-Bauer modified). The prepared compounds generally showed inhibitory activity against Gram+ bacteria. The greatest activity was observed in the compounds of general formula (B); they were especially inhibitory toward M. paratuberculosis ATCC 607.

Bacteria↗
PubMed2004

Simulation of alternatives for the Dutch Johne's disease certification-and-monitoring program.

To identify optimal method(s) for certification and subsequent monitoring of Mycobacterium avium subsp. paratuberculosis (Map)-unsuspected herds, certification-and-monitoring schemes were studied using a stochastic simulation model ("JohneSSim"). JohneSSim simulated the within-herd transmission and economic aspects of Map in closed Dutch dairy herds. The model was validated with field observations on Map-unsuspected herds. The current Dutch certification-and-monitoring schemes were compared with 11 alternative schemes in which individual and pooled fecal culture, ELISA, Johnin-intradermal test and gamma-IFN ELISA were used, varying the test frequency, tested age group and number of tested animals. On reaching the 'Map-free' status with the standard certification scheme, 11% of the simulated herds were not truly Map-free. Therefore, the designation 'Map-free' should be changed into, for instance, 'low-risk Map'. In the most-attractive alternative certification scheme, the 'Map-free' status was reached after four herd examinations (at 2-year intervals) consisting of serial testing of all cattle > or = 2 years of age with a pooled fecal culture and individual fecal culture of positive pools. This scheme resulted in lower total and annual discounted costs and a lower animal-level prevalence at reaching the 'Map-free' status compared to the standard scheme, assuming that there was no new introduction of the infection. Schemes to monitor the 'Map-free' status were compared, assuming that this status was reached with the standard certification scheme. In comparison to the standard monitoring scheme, none of the alternative monitoring schemes resulted in both a lower animal-level prevalence of undetected pre-existing Map infections in closed herds, and lower median annual discounted costs. Results of the model were very sensitive to the assumed sensitivity of the fecal culture test and to management measures that prevent within-herd transmission of Map. If these preventive measures were taken, the probability of undetected Map infections in closed 'Map-free' herds was decreased substantially.

Animal Husbandry↗