Biomedical subjects

A B Thiermann

Publications and source records attributed to A B Thiermann.

At least 19 recordsLinked to original sources

PubMed1991

Restriction endonuclease analysis as a taxonomic tool in the study of pig isolates belonging to the Australis serogroup of Leptospira interrogans.

Restriction endonuclease analysis was performed on DNAs from the type strains of the Australis serogroup of Leptospira interrogans by using 20 restriction enzymes, and the electrophoretic patterns obtained were compared with patterns obtained from 162 Australis serogroup isolates from pigs. It proved to be a quick and reliable method for typing such strains. All of the pig isolates were identified as either serovar bratislava or muenchen. It also showed differences at the subserovar level which may be important in (i) understanding the epidemiology of the Australis serogroup, (ii) the development of suitable vaccines, and (iii) pathogenesis and pathogenicity studies. Two genotypes (B2b and M2) accounted for 92% of isolates from aborted or stillborn piglets, while a third genotype (B2a) was the only one recovered from the brains of piglets with meningitis.

Animals↗

PubMed1989

Effect of vaccination with a pentavalent leptospiral vaccine on Leptospira interrogans serovar hardjo type hardjo-bovis infection of pregnant cattle.

Effectiveness of a pentavalent leptospiral vaccine to protect cattle from infection and reproductive problems caused by Leptospira interrogans serovar hardjo type hardjo-bovis was evaluated. Seven cows were vaccinated once and 8 cows were vaccinated twice with a USDA-licensed pentavalent leptospiral vaccine. Five cows were maintained as nonvaccinated controls. Cows were bred 1 to 2 months after the last vaccination. During the 4th to 6th month of gestation, all cows were challenge exposed on 4 occasions by conjunctival instillation of 10(8) serovar hardjo type hardjo-bovis organisms and on 3 occasions by conjunctival instillation of urine from a cow shedding hardjo-bovis. All control cows and 13 of 15 vaccinated cows became infected and shed leptospires in the urine. Leptospires were detected in fewer urine samples collected from vaccinated cows, compared with those collected from control cows. Four stillborn calves and 3 weak calves were born to control and vaccinated cows. Leptospires were detected in the kidneys of 11 apparently healthy calves born to vaccinated and control cows. Agglutinating antibodies were not detected in the precolostral serum of these calves.

Animals↗

PubMed1988

Leptospirosis.

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Animals↗

PubMed1988

The distribution of ferritin, lactoferrin and transferrin in granulomatous lymphadenitis of bovine paratuberculosis.

Immunohistochemical examination of iron-binding proteins was carried out in the formalin-fixed mesenteric lymph nodes of normal cattle and of cattle with paratuberculosis. Ferritin (FT) and lactoferrin (LF) were found in the granulomas in ileal lymph nodes from six infected cattle. A weak reaction for transferrin (TF) was found in granulomas of a lymph node from one of the infected cattle. FT was found in the macrophages in the medullary sinuses of normal and infected nodes; however, the reaction in infected nodes was generally stronger than that in normal ones. LF in the macrophages was found in only two infected nodes. Neutrophils in both normal and infected cattle always reacted strongly for LF. The TF was always found in the blood vessels and intracellular space. These results suggest that: (1) FT and LF may be important in vivo sources of iron for Mycobacterium paratuberculosis, since their own iron-binding compounds are considered to acquire iron from FT and LF in vitro; (2) the increase in FT and LF in the granulomas may be related to inflammatory hyposideraemia associated with paratuberculosis and (3) epithelioid and giant cells may have a different iron metabolism, from normal macrophages.

Animals↗

PubMed1988

Role of M cells and macrophages in the entrance of Mycobacterium paratuberculosis into domes of ileal Peyer's patches in calves.

Ligated ileal loops of calves were inoculated with live and heat-killed Mycobacterium paratuberculosis and were examined by light and electron microscopy. At 5 hours after inoculation, acid-fast bacilli were in subepithelial macrophages, but not in M cells covering domes. At 20 hours, more than 50 acid-fast bacilli per cross section were in subepithelial macrophages in domes. Both living and heat-killed bacilli passed into domes. Addition of anti-M. paratuberculosis bovine serum to the inoculum enhanced entry of bacteria into domes. By electron microscopy, intact bacilli with electron-transparent zones (peribacillary spaces) were in the supranuclear cytoplasm of M cells at 20 hours. M cells also contained vacuoles, including electron-dense material interpreted as degraded bacilli. Subepithelial and intraepithelial macrophages contained bacilli and degraded bacterial material in phagosomes. These results suggest that calf ileal M cells take up bacilli, and that subepithelial and intraepithelial macrophages secondarily accept bacilli or bacterial debris which are expelled from M cells.

Animals↗

PubMed1988

Restriction endonuclease analysis of Leptospira interrogans serovar hardjo isolates from cattle.

The genomes of 253 strains of Leptospira interrogans serovar hardjo were examined by restriction endonuclease analysis of their DNA. The strains had been isolated from cattle at an abattoir (190), milk of agalactic cows (seven) and from aborted bovine fetuses (56). Two distinct genome types, Hardjoprajitno and Hardjobovis, were detected. The majority (91 per cent) of isolates from abattoir cattle were of the Hardjobovis type while most (76 per cent) of the isolates from clinical/pathological material were similar to Hardjoprajitno.

Animals↗

PubMed1987

Genetic and antigenic differences of serologically indistinguishable leptospires of serovar hardjo.

Pathogenic leptospires of serovar hardjo isolated from North American cattle were compared genetically and antigenically to reference strain hardjoprajitno of the Sejroe serogroup. Guanine-plus-cytosine (G+C) contents were determined for the genomes, and microscopic agglutination, Western blotting (immunoblotting), and immunoprecipitation were used to characterize antigenic relatedness. Major differences were demonstrated between the isolates and the reference strain. The G+C content of the reference strain was calculated to be approximately 34 +/- 1%, and those of the isolates were calculated at 39 +/- 1%. Antigenic differences between the isolates and the reference strain were identified by using rabbit immune serum raised against a hardjo isolate exhaustively adsorbed with hardjoprajitno whole and sonicated cells. Western blot analysis and immunoprecipitation using this adsorbed serum revealed antigens apparently unique for the hardjo isolates. Microscopic agglutination with the adsorbed rabbit serum did not agglutinate hardjoprajitno when diluted 1:2 but agglutinated bovine isolates to a 1:32 dilution. Bovine antiserum raised against the isolates was also used to identify antigens by immunoprecipitation.

Agglutination Tests↗

PubMed1987

Isolation and analysis of restriction endonuclease digestive patterns of chromosomal DNA from Mycobacterium paratuberculosis and other Mycobacterium species.

A relatively rapid and efficient method for the extraction of chromosomal DNA from Mycobacterium paratuberculosis and other mycobacteria was developed. Approximately 25 to 50 micrograms of DNA could be extracted from 100 mg (wet weight) of cells, which was sufficient to perform several restriction endonuclease analyses from a single preparation. The DNA from five Mycobacterium species, including four strains of M. paratuberculosis and four strains of M. avium, was analyzed by this method. Digestion with the restriction endonucleases BstEII and PstI yielded the most definitive restriction patterns. For some strains, the restriction endonuclease analysis results were in agreement with the current identification of these organisms. The two strains of M. avium serotype 2 had identical fragment patterns. Similarly, the two strains of M. avium complex serotype 6 had identical fragment patterns. The three mycobactin-dependent M. paratuberculosis strains were very similar, whereas the mycobactin-independent M. paratuberculosis strain was more similar to the M. avium serotype 2 strains. Although many more cultures would need to be evaluated to determine correct groupings, the results of this study demonstrated the potential of restriction enzyme analysis for the differentiation of slowly growing mycobacteria.

Animals↗

PubMed1986

Survey of selected diseases in wild swine in Texas.

Tissue, fecal, and serum specimens and swabs of nasal turbinates and tracheas were collected from 100 wild swine (Sus scrofa) from 10 populations in Texas and, along with 24 additional serum specimens, were evaluated for selected swine diseases. Swine positive for pseudorabies were detected in 7 populations. Brucella suis biovar 1 was isolated from 4 swine from 2 populations, but positive serologic results may indicate a more widespread distribution of the organism. All populations contained swine that were positive for leptospirosis. Trichinella spiralis was not found in the swine evaluated.

Animals↗

PubMed1986

Isolation of leptospires from the genital tract and kidneys of aborted sows.

Twenty sows were examined as part of an investigation into the relative importance of the genital and urinary tracts as sites of leptospiral persistence in pigs. The sows had previously either aborted leptospire-infected fetuses or aborted on farms where leptospire infection had been demonstrated in other litters. Leptospires belonging to the Australis serogroup were recovered from the upper genital tract of all the sows. In contrast, leptospires were only recovered from 75 per cent of their kidneys. Isolates from 10 of the sows were identified to serovar level: six were serovar muenchen and four were bratislava. Persistence was observed in renal and genital tissues for up to 147 days after abortion.

Abortion, Veterinary↗

PubMed1986

Reclassification of North American leptospiral isolates belonging to serogroups Mini and Sejroe by restriction endonuclease analysis.

The genomes of North American strains of leptospires belonging to serogroups Mini and Sejroe were analyzed and compared with those of reference strains by cleavage with restriction endonucleases. The isolates selected for this study, when typed by the serologic method, were identified as serovars szwajizak, hardjo, and balcanica. However, the results of restriction endonuclease analysis (REA) indicated that a different classification existed. The 2 isolates typed as serovar szwajizak seem to be georgia by REA. Isolates belonging to serovars balcanica and hardjo had REA patterns that differed from both reference strains. Differences were not observed in the REA patterns between balcanica and hardjo isolates. All hardjo and balcanica isolates examined are suggested to be classified into a previously described hardjo, REA subtype hardjobovis. Using the enzyme Hha1, these isolates were subdivided into 3 subgroups. When examining the REA pattern of the 17 reference strains in serogroup Sejroe, 3 identical pairs were observed: wolffi and roumanica; sejroe and polonica; and istrica and nyanza. The REA again indicated that it will be a valuable method for the classification of leptospires.

DNA Restriction Enzymes↗

PubMed1986

Isolation of Leptospira interrogans serovar bratislava from sows in Iowa.

Leptospira interrogans serovar bratislava was recovered from 2 of 10 sows examined from an Iowa slaughterhouse. Isolations were made from the kidney and genital tract of each sow. Serovar bratislava is not included in vaccines because it has not been previously isolated in the United States.

Animals↗

PubMed1986

Isolation of leptospires from the genital tracts of Iowa cows.

Leptospira interrogans serovar hardjo was recovered from 5 of 11 nonpregnant cows chosen at random from an Iowa slaughterhouse. Isolations were made from the kidney in each instance, the genital tract of 3 cows: uterus from 1 cow, oviduct of another, and oviduct and uterus of a 3rd cow.

Animals↗

PubMed1986

DNA homology studies of leptospires of serogroups Sejroe and Pomona from cattle and swine.

Hybridization studies of chromosomal DNA from leptospiral strains representing Leptospira interrogans, serogroups Sejroe and Pomona from cattle and swine were performed to determine the degree of homology among their DNA sequences. Chromosomal DNA isolated from leptospires of the Sejroe and Pomona serogroups was radiolabeled and used to probe DNA isolated from other strains in these serogroups. Serovars hardjo (hardjoprajitno), hardjo (hardjo-bovis), balcanica (1627 Burgas), pomona (pomona), and kennewicki (LT-1026) were probed to determine the degree of homology among their chromosomes. Serovars pomona and kennewicki were homologous to each other. They also had a high degree of homology with hardjo (hardjoprajitno) and, to a lesser extent, with hardjo (hardjo-bovis) strains. However, hardjoprajitno and hardjo-bovis had little homology to each other. Serovar balcanica had a high degree of homology with hardjo-bovis isolates, but little homology with hardjoprajitno. Although serologically indistinguishable, the reference strain hardjoprajitno was genetically dissimilar to hardjo-bovis strains isolated from North American cattle.

Animals↗

PubMed1985

Identification and characterization of the protein antigens of Leptospira interrogans serovar hardjo.

We radiolabeled Leptospira proteins with [35S]methionine. Solubilized extracts of radiolabeled L. interrogans serovar hardjo strain hardjoprajitno were analyzed by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. We compared the protein profile obtained in this manner to the protein profiles of various [35S]methionine-labeled Leptospira spp. The profiles of the pathogenic L. interrogans strains were very similar but not identical and exhibited no obvious relationship to those of the two nonpathogenic species. We used solubilized, radiolabeled hardjoprajitno extracts and a sensitive radioimmunoprecipitation procedure to identify protein antigens recognized by immunoglobulin G antibodies present in various rabbit anti-hardjo sera. Homologous hyperimmune rabbit serum efficiently precipitated a large subset of proteins, the majority of which were between 30,000 and 66,500 daltons. Radioimmunoprecipitations with sera prepared against each of four recent hardjo isolates cultured from infected cattle produced similar results. Immunoprecipitations done with various radiolabeled Leptospira extracts and anti-hardjoprajitno serum demonstrated that the pathogenic leptospires possessed a number of cross-reactive major and minor protein antigens. By cell fractionation procedures, we found that most of the major protein antigens were present in the outer envelope. These proteins were exposed on the leptospiral cell surface because intact radiolabeled leptospires bound antibodies directed against them.

Antigens, Bacterial↗

PubMed1985

New method for classification of leptospiral isolates belonging to serogroup pomona by restriction endonuclease analysis: serovar kennewicki.

The genomes of leptospiral field isolates belonging to serogroup Pomona were analyzed and compared with those of type strains by cleavage with restriction endonucleases. This new classification method shows differences among these organisms not indicated by the conventional serological typing method. No differences were observed among isolates from the United States, Canada, and New Zealand. Although all isolates selected for this study had been serologically typed as belonging to serovar pomona, the restriction endonuclease analysis indicates that they belong to serovar kennewicki. kennewicki, a serovar of North American origin, has recently been eliminated from the official serovar list because it was found to be indistinguishable from serovar pomona by the serological method.

DNA Restriction Enzymes↗

PubMed1985

Rapid and simplified protocol for isolation and characterization of leptospiral chromosomal DNA for taxonomy and diagnosis.

We have developed a rapid method for the isolation of leptospiral chromosomal DNA which yields DNA of a purity suitable for restriction endonuclease analysis. A small volume (15 to 20 ml) of an exponentially growing culture of leptospires yielded 2 to 4 micrograms of chromosomal DNA. In a 1-day protocol, the DNA was isolated, restricted with endonucleases, and fractionated on an agarose gel. Chromosomal DNA from dinger zones (visible subsurface zones of leptospiral growth) of first semisolid subcultures of field isolates was also isolated and characterized, thus greatly speeding up the diagnostic process.

Chromosomes, Bacterial↗

PubMed1985

Experimental infection of calves with Leptospira interrogans serovar hardjo: conjunctival versus intravenous route of exposure.

Eight-month-old calves, housed under maximum isolation, were exposed to pathogenic Leptospira interrogans serovar hardjo by the conjunctival route or IV. One calf served as an unexposed control. Infection was monitored serologically (microscopic agglutination test and enzyme-linked immunosorbent assay; ELISA) and by leptospiral culture isolation from periodic urine samples and from the kidneys, epididymides, and aqueous humor collected at slaughter. Microscopic agglutination test titers of greater than or equal to 1:40 were detected among all IV exposed calves at postinoculation day (PID) 7 and among conjunctival exposed calves at PID 14. By ELISA, all IV exposed calves were positive by PID 3, whereas conjunctival exposed calves were positive at PID 14. The ELISA was more sensitive for the detection of antibodies against leptospires in cattle. Leptospires were isolated from the urine of 4 calves and from the kidney of 3 calves exposed by the conjunctival route, but not from IV exposed calves. The results indicated that the conjunctival route of exposure was a more natural and successful route for experimental infection of cattle with serovar hardjo.

Agglutination Tests↗