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Biomedical subjects

M Gottschalk

Publications and source records attributed to M Gottschalk.

At least 109 records · Page 6Linked to original sources
PubMed1993

Production of capsular material by Streptococcus suis serotype 2 under different growth conditions.

The procedure currently used for the production of Streptococcus suis antigen is very long and includes several subcultures. The aim of the present work was to study the in vitro production of capsular material by S. suis serotype 2 after each of these subcultures. The amount of capsular material produced was evaluated by electron microscopy using bacterial cells grown on blood-agar plates and in Todd-Hewitt broth (THB) or THB supplemented with serum. In addition, the production of antibodies in rabbits with antigens produced using different growth conditions was compared. Antigens produced after only three subcultures possessed as much capsular material as cells obtained after the complete procedure and induced a similar antibody response. The use of serum as a supplement to the broth did not assure a higher production of capsule; in addition, antibody titers obtained with antigens produced in THB were as high as those obtained with antigens produced in THB supplemented with serum. We recommend the use of three subcultures in nonsupplemented broth for the production of immunogens. This revised protocol offers two main advantages: it is less time-consuming because of the limited number of subcultures and is also less expensive since nonsupplemented broths are used.

Animals↗
PubMed1992

Studies on a murine model for evaluation of virulence of Streptococcus suis capsular type 2 isolates.

Five different parameters, time of incubation of the culture, type of culture medium, inoculum, strain of inbred mice, and age of mice, were tested using the LD50 technique to standardize a murine model for the evaluation of the virulence of Streptococcus suis capsular type 2 isolates. A model using 28 day-old mice belonging to CF1 strain appeared to give the best results. The inoculum size was the parameter most influencing the 50% lethal dose obtained with mice. Inoculation with 1-ml volume of a bacterial suspension instead of 0.1 or 0.5 ml decreased the LD50. The standardized model was used to evaluate the virulence of some isolates of known pathogenicity for pigs. The minimum lethal dose was used in the model and it appeared that the virulence of Streptococcus suis capsular type 2 isolates can be measured from highly virulent to totally avirulent.

Animals↗
PubMed1992

Production and characterization of two Streptococcus suis capsular type 2 mutants.

Two avirulent mutants of Streptococcus suis capsular type 2 (M2 and M42) were produced from a highly virulent strain. Mutant M2, obtained after serial subcultures of the parent strain in the presence of rabbit anti-capsular type 2 serum, no longer possessed the type-specific capsular antigen, as demonstrated by serotyping methods and immunoelectron microscopy. The Lancefield group D antigen could not be detected on the cell surface of this mutant using the immunogold labelling technique. SDS-PAGE of lysozyme treated cells demonstrated that a 44 kDa protein which was present in the parent strain, was absent in mutant M2. Immunoblotting using rabbit whole cell homologous anti-serum revealed that the protein was strongly immunogenic. Mutant M2 was totally avirulent in mice, and the homologous antiserum completely failed to protect mice against challenge with the parent strain. However, mutant M42, obtained after passages of the parent strain at 42 degrees C, remained capsulated but lacked the same 44 kDa protein as mutant M2. The quantity of sialic acid present in the capsule was similar to that of the parent strain. Despite the presence of antibodies against the capsule, antiserum prepared against M42 only partially protected mice against a challenge with the parent strain. The 44 kDa cell wall protein could act as a virulence factor as well as an important immunogen of S. suis capsular type 2.

Agglutination Tests↗
PubMed1992

Reduction in morbidity due to diarrhea in nursing beef calves by use of an inactivated oil-adjuvanted rotavirus-Escherichia coli vaccine in the dam.

An outbreak of neonatal diarrhea occurred among beef calves (2000 animals) from one large Argentinian farm in 1985. Rotavirus was detected in 78% (106/136) and enterotoxigenic Escherichia coli in 1.5% of the samples (2/136) obtained from sick calves. In comparison rotavirus was identified in only 1.6% (1/63) of the samples from clinically healthy calves. The rotavirus strain responsible for the outbreak was characterized as serotype 6 belonging to group A. In the following three years the protective capacity of a combined rotavirus-E. coli inactivated vaccine administered to the dams during the last third of the gestation period was evaluated on this farm by comparison of morbidity due to diarrhea in calves from vaccinated vs. placebo cows within the same year. The morbidity due to diarrhea among calves from dams in the vaccinated and placebo groups was 34% and 77%, respectively in 1986; 23% and 47% in 1987, and 15% and 34%, in 1988. In 1987 morbidity of diarrhea in calves born from vaccinated heifers was 54% and 74% in calves from placebo heifers. In 1988 morbidity from diarrhea was 41% and 54%, respectively among calves in these two groups. In all experiments, calves from heifers showed significantly greater morbidity than calves from cows. Differences in diarrhea morbidity between the vaccinated and placebo groups were statistically significant (P less than 0.05). Additional studies showed that the diarrhea had a significant influence (P less than 0.05) on the average live weight of the calves at weaning (5 to 7 months) with an average weight loss of 7.8 kg per calf among the calves affected with diarrhea.

Adjuvants, Immunologic↗
PubMed1992

Molecular analysis of isolates of Streptococcus suis capsular type 2 by restriction-endonuclease-digested DNA separated on SDS-PAGE and by hybridization with an rDNA probe.

This study was undertaken to assess the discriminatory value of restriction endonuclease (RE) digestion patterns of Streptococcus suis chromosomal DNA using polyacrylamide gel electrophoresis (SDS-PAGE) and DNA-rDNA hybridization. For the RE digestion patterns, DNAs were digested separately with the enzymes BamHI and BglII and the resultant fragments were separated by SDS-PAGE. An Escherichia coli rDNA probe derived from pKK3535 was used for the hybridization. Twenty-three S. suis capsular type 2 isolates recovered from diseased and clinically healthy pigs, from a human case, and from a cow were compared in this study. The majority of isolates associated with septicaemia belonged to one restriction endonuclease analysis (REA) profile group. Isolates associated with pneumonia belonged either to the REA profile group of isolates associated with septicaemia or to a second REA profile group. The REA profiles of isolates from clinically healthy animals were more heterogeneous. The REA profile of the type 2 reference strain, S735, which was originally isolated from a pig, was very different from those of the porcine and bovine isolates but similar to the profile of the human isolate. The profiles obtained after rDNA hybridization were more homogeneous. Although different patterns were detected in the 23 isolates, there was no correlation between the source of the isolate and the patterns observed with this technique.

Animals↗
PubMed1992

Antimicrobial resistance patterns and plasmid profiles of Streptococcus suis isolates.

Streptococcus suis isolates recovered from diseased animals in Quebec and western Canada and from human cases in Europe were tested for their susceptibility to different antimicrobial agents and screened for their plasmid content. Most isolates from Quebec were clindamycin, erythromycin, and tetracycline resistant; animal isolates from western Canada were notably less resistant to clindamycin and erythromycin, whereas human isolates were considerably more susceptible to most antimicrobials tested. More than 60% of isolates had plasmids that ranged from 1.5 to 35 kilobases (kb). Of the 7 plasmid profiles found, 2 were particularly frequent in isolates from Quebec and western Canada, suggesting the presence of epidemic strains in the swine population. A particular plasmid band of about 5 kb was present in most Canadian isolates. When this band was used as a probe in colony and Southern blot hybridization, most isolates harboring the 5-kb plasmid hybridized, even though their plasmid profiles were different. Human isolates from Europe differed in their plasmid content from Canadian isolates of animal origin. Although a high degree of antimicrobial resistance was associated with the presence of plasmids in most isolates, it was not possible to establish a causative relationship.

Animals↗
PubMed1991

Characterization and expression of a genomic pectin methyl esterase-encoding gene in Aspergillus niger.

The genomic pectin methylesterase (PME)-encoding gene (pmeA) from Aspergillus niger strain RH5344 was cloned by probing a genomic DNA library with a cDNA coding for PME. The recombinant phage clone was isolated and a 6-kb HindIII fragment was subcloned and characterized. The gene consists of seven exons and six introns. The nucleotide sequences of the coding regions were identical to those found in the pmeA cDNA. Cotransformation of A. niger was achieved with the vector, pAN7-1, and transformants were then tested for PME production. Transformants which produced more PME than the untransformed recipient strain were subjected to Southern-blot and Northern-blot analysis. The results show that there is a reasonable correlation between gene copy number, mRNA levels and PME production. PME was produced by A. niger transformants in an active 43-kDa form, which is similar to that of the mature protein isolated from the strain, RH5344. On the basis of the results of affinity labeling of PME with sugar-specific lectins and the amino acid sequence data, it has been revealed that PME is a glycoprotein and the protein-bound glycans are oligosaccharides with a high mannose content.

Affinity Labels↗
PubMed1991

Characterization of a polygalacturonase gene of Aspergillus niger RH5344.

We have cloned a gene encoding a polygalacturonase (PG) in the filamentous fungus Aspergillus niger RH5344. The structural gene comprises 1141 bp coding for 362 amino acids and the open reading frame is disrupted by one intron of 52 bp. Eukaryotic consensus sequences for transcription regulation are found only in deviated forms. The biological functionality of the isolated PG gene was established by retransformation in A. niger and Aspergillus awamori. In addition, we have found that the PG protein of A. niger shares significant similarities with PG proteins from tomato and Erwinia carotovora. Comparison of the three enzymes revealed a highly conserved region in their C-terminal region probably comprising the elements of substrate binding and the catalytic centre.

Amino Acid Sequence↗
PubMed1991

Characterization of six new capsular types (23 through 28) of Streptococcus suis.

Six new capsular types of Streptococcus suis (types 23 to 28) are described. All reference strains were isolated from diseased pigs and were morphologically and biochemically similar to previously described capsular types 1 to 22. Clear and specific reactions were obtained for each of the new capsular types with three different typing techniques; no cross-reactions were detected among them or with other S. suis capsular types. Their capsular material presented similar ultrastructural characteristics, as shown by electron microscopy, and fimbriae similar to those described for other capsular types of S. suis were observed. When untypeable field isolates were tested with antisera raised against the six new capsular types, capsular type 23 appeared to be the most prevalent, representing more than 50% of all these isolates. Most isolates were recovered from cases of pneumonia, septicemia, and meningitis. Presumptive biochemical identification described for S. suis capsular types 1 to 22 may also be used for capsular types 23 to 28.

Animals↗
PubMed1991

Isolation and characterization of Streptococcus suis capsular types 9-22.

The incidence and biochemical patterns of Streptococcus suis capsular types 9-22 are presented. Of 148 untypeable (with types 1-8 antisera) isolates of S. suis recovered from diseased pigs, 10% were not capsulated. Of the remaining 134 isolates, only 53% belonged to capsular types 9-22; capsular types 22 and 9 were the most prevalent, representing 19% and 13%, respectively. Capsular type 15 (de Moor's group T Streptococcus) is reported here for the first time in North America since it was described in 1963 in Europe. Of 188 untypeable isolates recovered from clinically healthy pigs, 25% were noncapsulated. Of the remaining 141 isolates, 90% belonged to the new capsular types, and 87% were identified as 1 of 4 types: 17, 18, 19, and 21. Capsular types 12 and 20 were not detected among the Canadian isolates. Almost half of strains were arginine dihydrolase-negative, and 45% fermented mannitol, which is seldom a positive test with capsular types 1-8. Although some strains were negative with salicin or trehalose, none were negative for both sugars. Only 54% of isolates tested with 1 rapid multitest system were correctly identified as S. suis. A tentative biochemical profile that might be used with a microplate identification system is also presented. Biochemical identification using the conventional system instead of the rapid multitest system is preferable.

Agglutination Tests↗
PubMed1991

Adherence of Streptococcus suis capsular type 2 to porcine lung sections.

The present study was undertaken to evaluate the ability of 33 Streptococcus suis capsular type 2 isolates to adhere to frozen sections of porcine lung. Twenty isolates originated from diseased pigs and 13 from the nasal cavities of clinically healthy pigs. All isolates from diseased animals adhered to lung sections; isolates from pneumonia adhered, in general, in greater numbers than isolates from meningitis. Only four isolates from clinically healthy animals showed a weak adherence to lung sections. Hydrophobic surface properties were also evaluated. All isolates tested appeared to possess a hydrophilic cell surface. The thickness of the capsular material correlated well with the degree of adherence. However, when the adherence capacity of a noncapsulated mutant was compared with that of the parent strain, it was found that the mutant strain had at least the same adherence capacity as the capsulated parent strain. The data suggest that S. suis capsular type 2 isolates involved in pathological conditions can adhere to porcine lung tissue. The adherence activity does not seem to involve hydrophobic interactions. The amount of capsular material seems to influence the adherence activity, but is probably not the only mechanism involved.

Animals↗
PubMed1991

[Comparative studies of in-vivo and in-vitro contraction behavior of gallbladders with impaired function].

The contractility of human gallbladders without calculi of 10 patients with typical biliary pain was investigated in vivo (n = 7; pharmacosonography) and in vitro (n = 10; isometric registration). In 6 out of 7 cases, investigated in vivo as well as in vitro, the contractility was identical--therefore the smooth-muscle-preparation in the organ-bath could be an interesting model for studies with CCK-antagonists. 3 of the in-vitro-preparations showed hypomotility consecutive to acetylcholine- and CCK-stimulation. Histologically, the destruction of the normal gallbladder wall structure by scars, chronic inflammation and the evidence of so-called Rokitansky-Aschoff-sinuses could be proved. The importance of these findings probably causal to the process of cholelithogenesis is discussed.

Acetylcholine↗
PubMed1990

Cloning and DNA sequence analysis of a polygalacturonase cDNA from Aspergillus niger RH5344.

A 1319 bp long cDNA encoding for a polygalacturonase (EC 3.2.1.15) from Aspergillus niger RH5344 comprises a single open reading frame of 1089 bp which includes the mature protein of 362 amino acids and an NH2-terminal signal peptide of 27 amino acids. The directly determined peptides of the mature polygalacturonase confirmed the sequence information deduced from the cDNA.

Amino Acid Sequence↗
PubMed1990

Ultrastructural study of surface components of Streptococcus suis.

The presence of capsular material on cells of nine reference strains of Streptococcus suis representing serotypes 1 to 8 and 1/2 was determined by transmission electron microscopy after polycationic ferritin labeling, immunostabilization, or fixation with a combination of glutaraldehyde and lysine. All the cells of the reference strains examined were covered with a layer of capsular material whose thickness varied between 20 to 30 nm and 350 to 375 nm when examined by immunostabilization. Capsular material from cells exposed to homologous antiserum was usually thicker than that from polycationic ferritin-labeled cells or cells fixed with glutaraldehyde-lysine. Negative staining revealed detectable surface structures on S. suis strains. All strains carried peritichous, thin, and flexible fimbriae with a diameter of approximately 2 nm and a length of up to 250 nm. This study indicated that morphological differences of surface structure exist among S. suis reference strains.

Ferritins↗
PubMed1990

Hemagglutination properties of Streptococcus suis.

A total of 49 strains (23 reference strains and 26 field isolates) of Streptococcus suis were tested for their ability to agglutinate erythrocytes from different animal species. Ten different hemagglutination patterns were established. Thirty-three strains (67%) did not agglutinate any of the erythrocytes tested; sixteen strains (33%) agglutinated erythrocytes from one or more animal species. Different strains belonging to the same capsular type presented different hemagglutination patterns. No correlation was found between the tissue origin and/or the virulence (evaluated in 4-week-old mice) of different field isolates and their hemagglutination activity. Hydrophobic surface properties were also evaluated. All S. suis strains studied appeared to possess a hydrophilic cell surface. Morphologically similar fimbriae were observed on hemagglutinating as well as on nonhemagglutinating strains of S. suis. This study provides evidence that certain strains of S. suis possess hemagglutinating properties which do not appear to involve hydrophobic interactions. The possible role of fimbriae in hemagglutination remains unclear.

Animals↗