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J F Timoney

Publications and source records attributed to J F Timoney.

At least 73 records · Page 4Linked to original sources
PubMed1991

Antigenic and genetic homogeneity of Streptococcus uberis strains from the bovine udder.

DNA- fingerprints (Hind III) of Streptococcus uberis field isolates from New York State and Europe showed substantial homogeneity, but were different to those of the type strain of the newly proposed psychrophilic species S. parauberis. S. uberis strains had major SDS-heat extracted antigens of molecular masses (Mr) less than 14, 40-41, 42-43, 59-61, 80-86 and 118-122 kDa following immunoblotting with rabbit hyperimmune sera. Bovine sera and milk reacted with the 40-41 and 118-122 kDa antigens. Variations in the Mr of particular bands were too unevenly distributed to permit formation of subgroups. Although cross reactive, the sizes of the antigens of S. parauberis strain NCDO 2020 were substantially different to those of S. uberis, the most prominent antigen having a Mr of 50 kDa. The antigenic and genetic data therefore strongly support the introduction of S. parauberis as a distinct species. S. uberis strains reacted with antiserum to Lancefield groups B, E, G and P, their grouping reactions showing no correlation with DNA and immunoblot fingerprints. Lancefield grouping of S. uberis therefore appears to have little value in identification.

Animals↗
PubMed1991

Characterization of a protective protein antigen of Erysipelothrix rhusiopathiae.

Although vaccination is widely practiced against infection by Erysipelothrix rhusiopathiae in pigs and turkeys, the protective antigen(s) involved have not been fully characterized or purified to homogeneity. Antigens of E. rhusiopathiae strain T28, serotype 2b, and of FRANKFURT XI, serotype N, in culture supernatant and in extracts made with hot acid, 10 mM NaOH, ultrasound or EDTA were compared by SDS-PAGE and immunoblotting and in a mouse protection test. EDTA and 10 mM NaOH yielded highly protective extracts; culture supernatant was less protective and ultrasonic or hot acid extracts stimulated little or no protection in mice. Protective antisera from swine, horses and mice recognized prominent bands of molecular mass (m.m.) of 66-64 and 40-39 kDa in EDTA and 10 mM NaOH extracts. Mice immunized with preparations of the 66-64 kDa band purified by preparative electrophoresis were protected. Both antigens were trypsin sensitive, contained no detectable polysaccharide, and showed a marked tendency to aggregate in the absence of SDS.

Animals↗
PubMed1991

A comparison of different methods of extraction of the M-protein from Streptococcus equi.

The molecular weights of the proteins produced in different extracts of Streptococcus equi were compared on immunoblots with antisera against acid extracted and mutanolysin extracted M-protein. Acid and alkaline extracts of S. equi contained some peptides of similar molecular weight that reacted with antiserum against an acid extracted 41,000 m.w. fragment suggesting that these fragments contained common epitopes. Comparison of the amino acid compositions of the 35,000 m.w. fragment of the alkaline extract and the 41,000 m.w. fragment of the acid extract suggest that these immunologically reactive fragments were probably derived from the same protein. Little cross-reactivity was observed between antisera against S. equi acid extracted protein and the native 58,000 m.w. M-protein. This suggests that conformational epitopes on the native M molecule are not present after acid treatment.

Amino Acids↗
PubMed1990

Cloning and expression in Escherichia coli of a protective antigen of Erysipelothrix rhusiopathiae.

Erysipelothrix rhusiopathiae is a primary pathogen of swine and turkeys and sporadic cause of disease in a variety of other hosts, including humans. A genomic library of the highly virulent strain of E. rhusiopathiae E1-6P was constructed in the expression-cloning vector lambda gt11 and screened with serum from a pig convalescent from an E. rhusiopathiae experimental infection. Immunoreactive clones were screened for their ability to protectively immunized mice. Two clones, lambda gt11/ersA and lambda gt11/ersB, were obtained that protected mice against challenge with E. rhusiopathiae E1-6P. Antisera against the recombinant clones reacted with polypeptides of molecular weights 66,000, 64,000, and 43,000 in detergent-solubilized surface antigen preparations and whole-cell lysates of E. rhusiopathiae. These polypeptides were also the major antigens recognized by convalescent pig serum when reacted with the same preparations. Western immunoblot and Southern blot analysis revealed that the cloned genes and gene products were present in all of the E. rhusiopathiae strains tested.

Animals↗
PubMed1990

Modified Feist broth as a serum-free alternative for enhanced production of protective antigen of Erysipelothrix rhusiopathiae.

The production of protective antigen in modified serum-free nutrient broth (H. Feist, K.-D. Flossmann, and W. Erler, Arch. Exp. Veterinaermed. 30:49-57, 1976) and in brain heart infusion broth supplemented with 10% horse serum (BHIS) was evaluated for six strains of Erysipelothrix rhusiopathiae serotypes 1a, 2, 2b, 4, and N. All six strains grew to higher cell densities in modified Feist medium than in BHIS and produced larger amounts of 64,000- to 66,000- and 39,000- to 40,000-molecular-weight antigens involved in immunity to erysipelas. A vaccine produced in Feist medium from E. rhusiopathiae SE-9 (serotype 2) was highly effective in a mouse protection test. We therefore suggest that modified Feist medium is an excellent, if not superior, alternative to BHIS for production of erysipelas vaccine.

Animals↗
PubMed1989

Clonal groups of Salmonella typhimurium in New York State.

The epidemiology of 278 strains of Salmonella typhimurium isolated from 1973 to 1981 from animals in New York State was studied by using four "fingerprinting" techniques, bacteriophage type (B.R. Callow, J. Hyg. 57:346-359, 1959), biotype (J. P. Duguid, E. S. Anderson, G. A. Alfredsson, R. Barker, and D. C. Old, J. Med. Microbiol. 8:149-166, 1975), plasmid profile, and antibiogram. Phage type with biotype was the most useful marker for distinguishing clonal groups of S. typhimurium. Four clones of S. typhimurium predominated, i.e., phage type/biotypes U275/26, 49/26, 10/3, and 2/3. U275/26 and 49/26 were commonly found until 1976, but clones 10/3 and 2/3 were predominant after 1976. Comparison of results with data from Canada suggested a dissemination of strains of S. typhimurium between Canada and New York. Cattle were a common source of phage type 49, as has been observed in other countries.

Animals↗
PubMed1989

Virulence determinants of Salmonella typhimurium from animal sources.

Two hundred seventy-eight strains of Salmonella typhimurium isolated from 1973 to 1981 from animal sources in New York State were studied for possible virulence determinants and for a serotype-specific plasmid possibly linked with virulence. Of the strains, 98% possessed type-1 fimbriae. All strains possessed flagella and were motile. One hundred twenty-three strains (44%) treated with mitomycin C tested positive for the cholera-Escherichia coli heat labile family of toxins by a kinetics-based ELISA; when treated with mitomycin C and extracted with polymyxin B, 249 (90%) were positive in the kinetics-based ELISA. All strains were negative in the Biken test. A smooth cell wall was found in 99% of the strains. Sixty-one percent (169) of the strains had a 62-Md plasmid. Seventy-six (27%) of the strains had detectable plasmids ranging in size from 1 to 124 Md.

Animals↗
PubMed1988

Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe.

A series of isolates of Streptococcus equi from the United States and Europe were compared by the bactericidal test, immunoblotting, DNA restrictions, and Southern hybridization analysis. All isolates tested were sensitive to the same bactericidal serum. In addition, immunoblotting revealed no differences in M proteins prepared by acid or mutanolysin extraction. Immunoblotting of acid extracts of the isolates with mucosal nasopharyngeal mucus from a convalescent horse revealed the presence of the 41,000- and 46,000-Mr polypeptide fragments of the M protein of S. equi known to be important in stimulating mucosal nasopharyngeal immune responses. DNA restriction analysis of total cell DNA digests, as well as Southern hybridizations using an S. equi M protein gene probe, did not detect any differences among these isolates. Our results, therefore, confirm the antigenic homogeneity of the M proteins of S. equi isolates and suggest that variation in this antigen is not a reason for the failure of commercial vaccines in the field. Interestingly, the protoplast M proteins of all isolates showed remarkable size homogeneity, in contrast to the size variation reported in M proteins of group A streptococci.

Animals↗
PubMed1988

Chemotactic response of equine polymorphonuclear leucocytes to Streptococcus equi.

Streptococcus equi infection in horses is characterised by intense infiltration of lymph nodes by polymorphonuclear leucocytes (PMNs) suggesting a potent chemotactic response to the organism or its products. Equine PMNs were separated using Ficoll-Hypaque medium and used in an assay of chemotaxis under agarose to study the components of S equi involved in this response. Results showed that complement-derived chemotactic factors generated by activation of the alternative complement pathway were important in chemotactic responses to S equi. Both whole bacteria and peptidoglycan preparations were potent complement activators, whereas purified M protein was less active. In contrast, S equi culture supernatant protein did not activate complement; instead it directly inhibited migration of PMNs. Moreover, PMNs, when incubated with culture supernatant of a non-haemolytic strain, showed signs of cellular degeneration suggesting the presence of a cytotoxin distinct from haemolysin.

Animals↗
PubMed1987

Molecular analysis of the M protein of Streptococcus equi and cloning and expression of the M protein gene in Escherichia coli.

A Streptococcus equi gene bank was constructed in the bacteriophage lambda gt11 cloning vector, and hybrid phage plaques were screened with S. equi M protein antiserum. A hybrid phage expressing the S. equi M protein (lambda gt11/SEM7) was identified and lysogenized into Escherichia coli Y1089. The cloned M protein appeared in immunoblots as three polypeptides with relative molecular weights of 58,000, 53,000, and 50,000. When reacted with S. equi M protein antiserum in an agar double-diffusion assay, the cloned M protein formed a line of identity with a protein in an acid extract of S. equi. Furthermore, lambda gt11/SEM7 protein inhibited opsonization of S. equi by antiserum to S. equi M protein. In addition, the recombinant protein expressed determinants of the antigen in the immune complexes of purpura hemorrhagica. Native M protein obtained from S. equi and recombinant M protein showed very similar molecular weight distributions on immunoblots, appearing as multiple closely spaced bands with molecular weights ranging from 52,000 to 60,000. Antisera prepared separately against each of the acid-extracted polypeptides shown to be important in serum bactericidal responses (molecular weight, 29,000) and nasopharyngeal local antibody responses (molecular weights, 41,000 and 46,000) of the horse each reacted with all three polypeptides in an acid extract. Moreover, antisera against protoplasts and against recombinant M protein of S. equi also reacted with these polypeptides. These results suggest that the entire M protein molecule of S. equi is present in these preparations and that the fragments in acid extracts carry overlapping segments.

Animals↗
PubMed1986

Passive transfer of mucosal antibody to Streptococcus equi in the foal.

Passive transfer of mucosal antibody to Streptococcus equi was studied in foals during the first 2 months of life. Immunoglobulin G (IgG) and IgA antibodies were found in sera and nasal secretions of foals shortly after colostrum intake. Titers were highest 2 days after birth; IgG predominated in sera, and IgA predominated in nasal washes. Intragastrically administered 99mTc-labeled IgA was transported from the bloodstream to the nasal mucosa of a newborn foal within a few hours of colostrum intake. Western blot analysis of the specificities of colostral and serum antibodies showed that selective transfer of immunoglobulins of defined specificity did not occur. Antibodies from milk samples taken a month or more into lactation had different specificities than those of colostrum or serum samples. Acid-extracted M protein fragments of S. equi recognized by milk antibodies were the same as those recognized by IgG and IgA from nasopharyngeal mucus of horses recently recovered from strangles. We postulate that passive antibody protection of the foal is derived both by secretion of colostral immunoglobulins onto the nasopharyngeal mucosa and by immunoglobulins ingested in milk that directly coat the upper respiratory and oral mucosa during the first months of life.

Age Factors↗
PubMed1986

Salmonella serotypes from animals in New York State, 1978-1983.

The salmonella serotypes isolated during 1978 to 1983 at the diagnostic and clinical laboratories of the New York State College of Veterinary Medicine from animal sources in New York state were reviewed and compared to earlier data from New York state animals and to national data for both human and animal sources. A total of 255 salmonella strains were studied from the six year period and included 33 serotypes. Salmonella enteritidis ser Typhimurium continued to be the most commonly reported serotype followed by serotype Anatum. Generally serotypes from New York state animals reflected national data on salmonella serotypes with a few exceptions. Serotype Dublin continued to be absent from New York animal populations. Bovine and equine species were the most common salmonella sources.

Animals↗
PubMed1986

Characteristics of an R antigen common to Streptococcus equi and zooepidemicus.

An R antigen of the group C streptococcus S. equi that cross reacts with a similar antigen of S. zooepidemicus has been identified and characterized. It is acid, heat and trypsin resistant, but pepsin sensitive and has an isoelectric point of 4.8. The amino acids in highest concentration are glutamic, aspartic, alanine, leucine, and valine. Bacterial components released in a French Press contain large amounts of R antigen, which is present also in culture supernatants and acid extracts. It has a molecular weight of about 82,000. Trypsin extraction of cells yields molecules of predominantly 56,000 and 25,000 molecular weight that appeared by immunoblotting to be similar to those obtained by trypsinization of purified R protein from preparations derived from the French Press. Although horses naturally infected with S. equi or S. zooepidemicus develop cross reacting R antibodies, the role of the R antigen in pathogenesis is unknown. Purified R protein does not stimulate bactericidal antibody in horses nor is it protective for mice. Its occurrence in antigen preparations in assays for S. equi antibody could be a source of interpretive errors because of the presence in many sera of antibody to the immunologically similar R antigen of S. zooepidemicus, a normal nasopharyngeal commensal of Equidae.

Animals↗
PubMed1985

Serum bactericidal responses to Streptococcus equi of horses following infection or vaccination.

An indirect test based on horse blood was used to study bactericidal responses of the horse to Streptococcus equi following infection or vaccination. Bactericidal antibody appeared in convalescent sera between two and four weeks and high titres were usually attained by eight weeks. Infection without clinical evidence of abscessation was also effective in eliciting strong bactericidal responses. Serum bactericidal activity of horses either recovered from strangles or immunised with commercial bacterin had declined eight months after vaccination. However, horses that developed strangles eight to 10 months after vaccination exhibited rapid and substantial increases in serum bactericidal activity. Groups of yearlings immunised with commercial S equi vaccines consisting either of M protein or bacterin developed clinical strangles within six months of vaccination although the majority of the animals had exhibited strong serum bactericidal activity a few weeks before occurrence of the disease. Similarly, a group of seven yearling ponies hyperimmunised with experimental vaccine, rich in M protein, were found to be highly susceptible to an intranasal challenge of 5 X 10(8) colony forming units of S equi, although their sera exhibited strong bactericidal activity at the time of challenge. These observations suggest that the role of serum bactericidal antibody in protection of the horse against strangles has been overrated.

Animals↗
PubMed1985

Mucosal nasopharyngeal immune responses of horses to protein antigens of Streptococcus equi.

Mucosal nasopharyngeal immunoglobulin A (IgA) and IgG responses to proteins of Streptococcus equi were studied in horses after the experimental production of strangles. S. equi-specific IgA and IgG titers in nasopharyngeal mucus were much higher in samples from animals 1 to 2 weeks after challenge than in samples from control animals. Although IgA was the major immunoglobulin in nasal mucus, there was more antibody activity associated with IgG as measured by radioimmunoassay. Great differences between the specificities of antibodies in nasal mucus and in serum were detected. IgA and IgG of mucus origin recognized only two major proteins with molecular weights of about 41,000 and 46,000 in acid extracts of S. equi and gave no detectable reaction with culture supernatant proteins. Only one protein of about 62,000 molecular weight was recognized in acid extracts of an equine strain of S. zooepidemicus. In contrast, immunoglobulins in serum recognized a great variety of proteins in culture supernatants and acid extracts of S. equi and S. zooepidemicus which did not include those proteins recognized by immunoglobulins in mucus. These findings provide good evidence for the independence of the local and systemic immune responses of the horse to S. equi. Horses rechallenged shortly after recovery from the first infection were resistant to challenge with an inoculum of S. equi 10 times greater than that to which they were originally susceptible. This resistance appeared to be independent of the levels of bactericidal antibody in serum. We therefore suggest that immunity to S. equi infection is mediated by locally produced nasopharyngeal antibodies.

Animals↗