Biomedical subjects

M Jacques

Publications and source records attributed to M Jacques.

At least 73 records · Page 4Linked 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

Affinity for porcine respiratory tract mucus is found in some isolates of Actinobacillus pleuropneumoniae.

The ability of 17 Actinobacillus pleuropneumoniae isolates representing serotypes 1, 2, 5, and 7, to adhere in vitro to porcine respiratory tract mucus was examined. Adherence of bacteria to crude mucus preparations was evaluated by use of a dot-blot assay and an enzyme immunoassay. Seventy per cent (12/17) of the isolates of A. pleuropneumoniae had affinity, to various degrees, for porcine respiratory tract mucus. No relationship was found between affinity for respiratory mucus and serotype, haemagglutination, lipopolysaccharide (LPS) profiles, or adherence to porcine tracheal rings. However, a correlation was found between affinity for respiratory mucus and capsular material thickness; heavily encapsulated isolated showed no or less affinity for mucus than isolates with a thinner layer of capsular material. Moreover, two encapsulated isolates showed less affinity for mucus than their acapsulated variant. Finally, the affinity of A. pleuropneumoniae for respiratory mucus was heat- and proteinase-K-resistant. Our data suggest that capsular material of A. pleuropneumoniae could mask a surface component, possibly LPS, which has affinity for porcine respiratory mucus.

Actinobacillus Infections↗

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

Cell surface characteristics and virulence in mice of Pasteurella multocida.

The virulence of three avian strains of Pasteurella multocida was evaluated in mice. Strains P-1059I (serotype A:3) and its uncapsulated variant P-1059B and strain 2723 (serotype A:16) were compared. Capsular material thickness after polycationic ferritin labelling of dextrose starch agar (DSA)-grown P. multocida was shown to vary with the strain and was not always related to virulence. Addition of alpha,alpha' bipyridyl (BIP) (160 microM) to the culture medium did not affect capsule production but increased virulence of strains P-1059B and 2723. None of the strains tested showed dermonecrotic activity. Using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), outer membrane protein (OMP) profiles indicated for strains P-1059I and P-1059B three proteins of 30, 35, and 38 KDa with the 30 KDa protein being the major one. Strain 2723 showed the same OMP profile but the 38 KDa protein was the major one. DSA + BIP-grown strains showed the same OMP profiles. Whole cell profiles were similar for all strains tested. However, addition of BIP to the culture media increased the virulence of strains P-1059B and 2723 and for all strains a 39 KDa protein was induced by the iron chelator. The results indicate that encapsulation may be important for virulence, but other surface components such as OMPs may be required as well.

Animals↗

PubMed1992

Isolation and characterization of adhesin-defective TnphoA mutants of septicaemic porcine Escherichia coli of serotype O115:K-:F165.

Non-enterotoxigenic porcine Escherichia coli strains belonging to the serogroup O115 have been associated with septicaemia and diarrhoea. Putative factors important in the pathogenicity of E. coli of serogroup O115 include fimbrial antigen F165, haemagglutination (MRHA), lipopolysaccharide, serum resistance, capsule and production of aerobactin. Using TnphoA transposon insertion mutagenesis, two classes of mutants were obtained from E. coli of serotype O115:F165 with respect to the phenotypic expression of fimbrial antigen F165 and MRHA of sheep erythrocytes: class I, F165-MRHA-, serum resistant; class II, F165+MRHA-, serum resistant. In a chicken lethality model, class I mutants were either virulent or of intermediate virulence, while class II mutants were of intermediate virulence. Alkaline phosphatase activity of class I and class II TnphoA mutants showed similar environmental regulation to that of fimbrial antigen F165. Moreover, class I and class II mutants were mutated in the prs-like locus, and lacked a 18.5 kDa and/or a 17.5 kDa fimbrial band.

Adhesins, Escherichia coli↗

PubMed1992

Molecular cloning of a determinant coding for fimbrial antigen F165(1), a Prs-like fimbrial antigen from porcine septicaemic Escherichia coli.

The genetic determinant coding for F165(1) fimbriae was cloned from the chromosome of the porcine Escherichia coli wild-type strain 4787 (O115:K-:H51:F165). The fimbrial determinant was further subcloned into the BamHI site of pACYC184 and a restriction map was established. On Southern hybridization, identity between the chromosomally encoded prs-like determinant of strain 4787 and its cloned counterparts was demonstrated. The cloned F165(1) fimbriae and those of the wild-type strain possessed a major protein subunit of molecular mass 18.5 kDa. Strains expressing F165(1) fimbriae were detected using an F165-specific polyclonal antiserum and caused mannose-resistant haemagglutination and agglutination of Forssman latex beads. Antiserum against the cloned F165(1) fimbriae recognized a 18.5 kDa band in the parent strain 4787.

Amino Acid Sequence↗

PubMed1992

Effects of sub-MICs of antibiotics on cell surface characteristics and virulence of Pasteurella multocida.

The effects of sub-MICs of certain antibiotics, namely, penicillin G, tetracycline, and trimethoprim-sulfamethoxazole, on the cell surface characteristics and the virulences of two toxigenic isolates of Pasteurella multocida representing capsular types A and D were evaluated. Expression of proteins, in particular, outer membrane proteins and iron-regulated proteins, was not affected by exposure of bacterial cells to low concentrations of antibiotics. However, exposition of surface antigens was modified by sub-MICs of the antibiotics tested. The lipopolysaccharide profile of one isolate (capsular type D) was altered by penicillin G. Sub-MICs of penicillin G and tetracycline diminished the virulence of the capsular type A isolate and adherence to porcine tracheal rings of the capsular type D isolate. Production of dermonecrotic toxin was not affected by sub-MICs of the antibiotics tested. Our results indicate that growth of P. multocida in the presence of low concentrations of antibiotics seems to have, depending on the isolate, profound effects on cell surface characteristics, with concomitant effects on adherence or virulence. Our results also indicate that production of dermonecrotic toxin, an important virulence factor of P. multocida isolates associated with porcine atrophic rhinitis, was not affected by sub-MICs of the antibiotics studied.

Animals↗

PubMed1992

Modification in penicillin-binding proteins during in vivo development of genetic competence of Haemophilus influenzae is associated with a rapid change in the physiological state of cells.

By using whole-cell labeling assay with 125I-penicillin V, we observed a reduction in the binding of the radiolabeled beta-lactam to four or five penicillin-binding proteins (PBPs) in Haemophilus influenzae cells cultivated under specific conditions. PBPs 3A, 3B, 4, and 6 were altered after the growth of bacteria in diffusion chambers implanted in the peritoneal cavity of rats. PBP 2 was also modified when cells were cultivated in human cerebrospinal fluids. Because this observation may have important consequences on the efficacy of beta-lactams during antibiotic therapy, we characterized the physiological state of bacteria cultivated in animals in the hope of explaining how such important changes in cell properties develop in vivo. Since the development of natural genetic competence occurs at the stationary phase of growth in H. influenzae, we used a DNA transformation assay to evaluate the physiological state of bacteria grown in diffusion chambers implanted in rats. Chromosomal DNA isolated from an antibiotic-resistant donor strain was mixed with bacteria in diffusion chambers. At different times during a 5-h incubation period, recipient bacteria were collected from the chambers, CFU were determined by plate counting, and antibiotic-resistant transformants were isolated on selective plates. Genetic competence rapidly developed in cells grown in rats, and the frequency of transformation by test DNA was elevated. Electron microscopy revealed an irregular cell shape and blebs at the surface of bacteria cultivated in animals and in cerebrospinal fluids. In an attempt to induce a similar physiological state in vitro, we supplemented broth cultures with cyclic AMP or synchronized cultures by a nutritional upshift. No changes in PBPs were observed with supplemental cyclic AMP or during a single cell cycle. Finally, a reduction in the affinity of PBPs for 125I-penicillin V identical to that observed in bacteria grown in rats was observed in cells isolated from the stationary phase of growth in vitro. These results clearly indicate that H. influenzae cells grown in animals undergo a rapid change to a physiological state similar to that found in late-stationary-phase cultures in vitro. This observation indicates that the rational design of future and improved antibiotic therapy of H. influenzae infections should consider cell properties of slow-growing or latent bacteria.

Animals↗

PubMed1992

Characterization of a polysaccharide capsular antigen of septicemic Escherichia coli O115:K "V165" :F165 and evaluation of its role in pathogenicity.

Escherichia coli strains of serogroup O115:K(-):F165 have been associated with septicemia in calves and piglets. These strains express a capsular antigen referred to as K"V165" which inhibits agglutination of the O antigen by anti-O115 serum. We used hybrid transposon TnphoA mutants M48, 18b, and 2, and a spontaneous O-agglutinable mutant, 5131a, to evaluate the role of K"V165" in the pathogenicity of E. coli O115. Mutant M48 was as resistant to 90% rabbit serum and as virulent in day-old chickens as the parent strain 5131, mutants 18b and 5131a were less resistant to serum and less virulent in chickens, and mutant 2 was serum sensitive and avirulent. Analysis of outer membrane protein and lipopolysaccharide profiles failed to show any difference between the transposon mutants and the parent strain. In contrast, the spontaneous O-agglutinable mutant showed additional bands in the 16-kDa region of the polysaccharide ladder-like pattern. Mutants 2 and 5131a produced significantly less K"V165" capsular antigen than the parent strain, as demonstrated by a competitive enzyme-linked immunosorbent assay with adsorbed anti-K"V165" serum. In addition, electron microscopic analysis revealed that mutants 2 and 5131a had lost the capsular layer observed in the parent strain after fixation with glutaraldehyde-lysine. This capsule contained carbohydrate compounds and resembled an O-antigen capsule since it prevented O-antigen agglutination before the bacteria were heated at 100 degrees C and induced bacterial serum resistance. The capsule-defective mutants colonized the intestinal epithelium of experimentally infected gnotobiotic pigs but failed to induce clinical signs of septicemia. We concluded that E. coli strains of serogroup O115 expressed a polysaccharide capsular antigen which induced serum resistance and consequently contributed to the pathogenicity of the bacteria.

Animals↗

PubMed1992

Molecular characterization of Serpulina (Treponema) hyodysenteriae isolates representing serotypes 8 and 9.

The study described here was carried out to further characterize reference strains of Serpulina (Treponema) hyodysenteriae representing serotypes 8 and 9. Results obtained from restriction fragment length polymorphism analysis, enteropathogenicity testing, and endotoxin profiles confirmed their identifications. Electron microscopy indicated that both strains were covered with a thin layer of capsule-like material. Immunoblot analysis indicated that an antigen in the 19-kDa region of proteinase K-digested whole cells reacted only with homologous antiserum. The serotype-specific antigens were sensitive to periodate oxidation but resistant to proteinase K digestion and migrated in the same region as purified lipopolysaccharides. Immunoblotting with proteinase K-digested whole cells appeared as useful as immunodiffusion with extracted lipopolysaccharide for the serological classification of S. hyodysenteriae. Immunogold labeling of whole cells and purified periplasmic flagella showed strong cross-reactions between S. hyodysenteriae and Serpulina innocens. Outer membrane preparations of strains representing serotypes 8 and 9 contained four major proteins which reacted with antisera against both species, and one major protein with a molecular mass of 46 kDa which reacted only with antisera against S. hyodysenteriae, irrespective of the serotype. Our findings suggest that periplasmic flagella and some outer membrane proteins are antigens common to both S. hyodysenteriae and S. innocens, whereas a 46-kDa outer membrane protein may be a species-specific antigen of S. hyodysenteriae. Finally, we propose immunoblotting as an alternative method to immunodiffusion for the serotyping of S. hyodysenteriae.

Animals↗

PubMed1992

In vitro colonization of porcine trachea by Mycoplasma hyopneumoniae.

Porcine tracheae maintained in culture were used in order to study the colonization by Mycoplasma hyopneumoniae. Rings excised from tracheae of newborn piglets were infected with M hyopneumoniae strain BQ 14 and, after different incubation times, were examined by light and electron microscopy. Non-infected tracheal mucosae maintained a normal appearance for several days. Infected tracheal rings showed progressive colonization with concomitant progressive damage to the mucosal surface. Early on during the infection, few mycoplasmas occurred over a ciliated epithelium. As the infection progressed, there was gradual loss of cilia; mycoplasmas tended to form microcolonies and to accumulate over the remaining ciliated cells. Mycoplasmas, first seen at the apex of the cilia, were then seen deeper in the inter-ciliary space; some were even seen in contact with microvilli. In histological investigation, the final stage of the infection was characterized by a marked destruction of the epithelium with exfoliation of the epithelial cells. Infected mucosae showed typical damage caused by M hyopneumoniae, namely reduction of ciliary activity after 5 days, loss of cilia, and sloughing of ciliated cells. Our data indicate that porcine tracheal organ culture can be advantageously used to study colonization by M hyopneumoniae.

Animals↗

PubMed1992

Enhanced adherence of Pasteurella multocida to porcine tracheal rings preinfected with Bordetella bronchiseptica.

Adherence of 25 isolates of Pasteurella multocida to porcine tracheal rings was evaluated. Results indicated that adherence was not related to the isolate's origin, capsular or somatic types, dermonecrotoxin production or hemagglutination activity. The effect of a preinfection with Bordetella bronchiseptica on the colonization by P. multocida was then studied. On rings infected with P. multocida alone, bacteria initially adhered to the epithelium, but within a few hours, the level of colonization decreased progressively. On rings preinfected with B. bronchiseptica, or pretreated with a cell-free B. bronchiseptica culture supernate (or filtrate), a high level of P. multocida colonization was maintained for at least 24 hours. Results indicate that B. bronchiseptica appears to facilitate upper respiratory tract colonization by P. multocida by a process which involves a low molecular weight (less than or equal to 1000) heat-stable substance, possibly the tracheal cytotoxin.

Animals↗

PubMed1991

Adherence of Actinobacillus pleuropneumoniae to porcine tracheal epithelial cells and frozen lung sections.

The ability of 23 different Actinobacillus pleuropneumoniae isolates to adhere in vitro to porcine tracheal epithelial cells and to porcine frozen lung sections was examined. It was found that A. pleuropneumoniae adhered poorly to isolated tracheal epithelial cells. On the other hand, A. pleuropneumoniae adhered to frozen lung sections and marked variations were observed between and within serotypes. Adherence to lung sections did not seem related to the hemagglutinating activity of the isolate. Two noncapsulated variants adhered to lung sections in greater numbers than their capsulated parent strains. Adherence to lung sections was not inhibited by the extracellular matrix components tested namely, laminin, fibronectin, and collagen, but was inhibited by homologous serotype-specific antiserum. The data indicated that the A. pleuropneumoniae isolates tested possess the ability to adhere to porcine lung tissue, a property which did not seem to be related to the serotype and did not seem to involve the capsular material or the hemagglutinins of the isolates.

Actinobacillus↗

PubMed1991

Geographic distribution and inheritance of three cytoplasmic incompatibility types in Drosophila simulans.

Wolbachia-like microorganisms have been implicated in unidirectional cytoplasmic incompatibility between strains of Drosophila simulans. Reduced egg eclosion occurs when females from uninfected strains (type W) are crossed with males from infected strains (type R). Here we characterize a third incompatibility type (type S) which is also correlated with the presence of Wolbachia-like microorganisms. Despite the fact that the symbionts cannot be morphologically distinguished, we observed complete bidirectional incompatibility between R and S strains. This indicates that the determinants of incompatibility are different in the two infected types. S/W incompatibility is unidirectional and similar to R/W incompatibility. A worldwide survey of D. simulans strains showed that type S incompatibility was found only in insular populations which harbor the mitochondrial type SiI. Both W and R types were found among mainland and island populations harboring the worldwide mitochondrial type SiII. Type S incompatibility could be involved in the reinforcement of the geographical isolation of SiI populations.

Animals↗

PubMed1991

Effects of antibiotics on the growth and morphology of Pasteurella multocida.

The effects of subminimal inhibitory concentrations (subMICs) of certain antibiotics, namely penicillin G, tetracycline and trimethoprim/sulphamethoxazole, on the growth and morphology of Pasteurella multocida were evaluated. SubMICs of penicillin markedly reduced the growth of P. multocida. Tetracycline and trimethoprim/sulphamethoxazole had no effect on its growth. SubMICs of penicillin greatly affected the morphology of P. multocida. At the highest concentrations tested (1/2 and 1/4 MIC) cells were acapsulate, and long filamentous cells (4-6 microns) were observed with some isolates. There was no correlation between the observed differences in the penicillin-binding proteins of the P. multocida isolates, and the extent of cell filamentation induced by penicillin G. SubMICs of tetracycline and trimethoprim/sulphamethoxazole did not seem to affect capsule production although filamentation was observed. Our results indicate that subMICs of penicillin can reduce growth of P. multocida. Furthermore, results also indicate that subMICs of antibiotics can affect the production of capsular material and the morphology of P. multocida.

Animals↗

PubMed1991

Experimental infection of newborn pigs with an attaching and effacing Escherichia coli O45:K"E65" strain.

The ability of a nonenterotoxigenic, K88-negative porcine Escherichia coli strain of serogroup O45:K"E65" to induce attaching-effacing lesions was investigated in newborn pigs. Typical attaching-effacing lesions, characterized by intimate adherence of bacteria to mature enterocyte brush borders with effacement of the microvilli, were observed on light and electron microscopy. Bacteria were also seen in intracytoplasmic vacuoles of mature enterocytes and, in areas of heavier colonization, in the lamina propria of the intestinal mucosa. A moderate inflammatory response with mild focal ulceration of the intestinal mucosa was observed. In a sequential study, we observed that the attaching-effacing lesions were well established in the duodenum, jejunum, and ileum at 12 h postinoculation but did not develop in the cecum and colon until 24 to 48 h postinoculation, although bacteria had colonized the latter areas as early as 12 h postinoculation. Initially, bacteria were very intimately attached, with an irregular arrangement on the enterocyte apical cell membrane, and subsequently reoriented to form a typical palisade arrangement with a narrow regular gap between the bacterial cell wall and the enterocyte apical cell membrane. This phenomenon of early intimate attachment of irregularly disposed bacteria has not been reported for human enteropathogenic attaching and effacing E. coli and could represent a new and different mechanism of attachment and effacement to intestinal epithelial cells.

Animals↗

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

Serotyping of Canadian isolates of Treponema hyodysenteriae and description of two new serotypes.

A total of 30 isolates of Treponema hyodysenteriae collected in the Saint-Hyacinthe (Quebec, Canada) area were serotyped by agar gel double immunodiffusion by using extracted lipopolysaccharide and hyperimmune rabbit antisera. Only 17% (5 of 30) of the isolates were typed with antisera specific for each of the seven known serotypes of T. hyodysenteriae. Antisera raised against 11 untypeable local isolates were then produced and tested against each lipopolysaccharide extract. Results showed two serologically distinct groups among 21 of the 25 untypeable isolates. The isolates in each group shared identical antigens. No detectable reactions could be observed between antisera raised against these 11 isolates and the antigens extracted from 7 reference serotype strains. On the basis of these results, two new serotypes of T. hyodysenteriae, serotypes 8 and 9, are proposed. We also propose isolate FM 88-90 as the reference strain for serotype 8 and isolate FMV 89-3323 as the reference strain for serotype 9. These two new serotypes, which represented 70% of the isolates tested, seem to be the major serotypes found in the province of Quebec.

Animals↗