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Biomedical subjects

C H Armstrong

Publications and source records attributed to C H Armstrong.

At least 19 recordsLinked to original sources

Relationship among selected Leptospira interrogans serogroups as determined by nucleic acid hybridization.

Leptospiral DNAs from a variety of Leptospira interrogans serogroups of veterinary significance, as well as a nonpathogenic leptospira, were compared by Southern blot hybridization of EcoRI-digested genomic DNA. The serogroups examined could be assigned to one of three groups on the basis of the degree of cross-hybridization between genomic DNAs. Only a few restriction fragments hybridized between the three groups, and most of these were shown to contain ribosomal DNA. The restriction fragment length polymorphism observed among the intergroup hybridizations allowed differentiation among serogroups and, in some cases, serovars. Under the hybridization conditions used, no hybridization was observed between leptospiral DNA and Leptonema, Escherichia coli, or porcine DNA.

Animals↗

Systemic salmonellosis in mature beef cows.

Systemic infection of mature beef cows with Salmonella typhimurium resulted in death of cows, abortions, and premature births. Salmonella typhimurium was isolated from the kidney, liver, and spleen of cows but not from an aborted fetus. Diarrhea was not a prominent clinical feature of the epizootic. The source of the salmonella was not determined.

Acute Disease↗

Etiologic studies on late-term swine abortions.

One hundred thirty-eight swine abortions were studied in detail in an effort to identify an etiologic agent. A viral agent was implicated in 7 cases. Bacteria were isolated in less than half of the examined cases: however, in 61% of the cases, motile, filamentous organisms were observed in tissues and fluids. Although swine sera from farms experiencing reproductive problems had a high reactor rate to Leptospira bratislava antigen, electron microscopy of the observed organism revealed a wall-free prokaryote morphologically typical of the class Mollicutes.

Abortion, Veterinary↗

Serological, pathological and cultural evaluations of swine infected experimentally with Mycoplasma flocculare.

Fourteen caesarean-derived, colostrum-deprived pigs and seven conventional swine were exposed to low passage, cloned, field isolates of Mycoplasma flocculare. Sera were collected at varying intervals postexposure (PE) and tested against M. flocculare and M. hyopneumoniae antigens in a semi-automated ELISA. Swine were killed six to 17 weeks PE and their lungs examined grossly for lesions and culturally for mycoplasmas. Pure cultures of M. flocculare were recovered from the lungs of 11 of 14 swine killed six to 12 weeks PE. Mycoplasmas were not isolated from the swine killed 15 to 17 weeks PE. Only one pig had gross lesions of pneumonia. Immunoassays revealed that swine were slow to seroconvert and titers (expressed in terms of optical density) were low. Three of 21 swine had antibodies to M. flocculare five weeks PE, five of 17 had seroconverted at seven to eight weeks and all surviving swine had antibodies to M. flocculare 76 days PE and beyond. Net optical density of positive sera was in the range of 0.201 to 0.412 (an optical density of 0.2 regarded as the breakpoint between negative and positive reactions in our ELISA). All of the sera were ELISA-negative when tested against M. hyopneumoniae antigen. This is regarded as a very significant finding. There has been concern that field sera might contain antibodies to M. flocculare and that such antibodies could render serodiagnostic tests for mycoplasmal pneumonia of swine nonspecific. Results of the present study suggest that swine infected with M. flocculare do not develop sufficient levels of antibodies to interfere with enzyme immunoassays for M. hyopneumoniae.

Animals↗

Cross-reactions between Mycoplasma hyopneumoniae and Mycoplasma flocculare--practical implications for the serodiagnosis of mycoplasmal pneumonia of swine.

Previous studies using hyperimmune antisera revealed significant serologic cross-reactions between Mycoplasma hyopneumoniae and M. flocculare. These findings, coupled with observations that M. flocculare infection may be widespread, indicated that antibodies to M. flocculare might be common in swine sera and thus cause nonspecific reactions in serodiagnostic tests for mycoplasmal pneumonia of swine (MPS). Information reported here indicates that this premise may be invalid. Twenty-one swine were infected experimentally with M. flocculare. Sera were collected every 2 weeks and tested against M. flocculare and M. hyopneumoniae antigens in an enzyme-linked immunosorbent assay (ELISA). Immunoassays against M. flocculare revealed that swine were slow to seroconvert, and that seroconversion was characterized by low titers. Significantly, none of the sera reacted with M. hyopneumoniae. M. hyopneumoniae reference antisera, and sera from swine naturally infected with M. hyopneumoniae and M. flocculare were also evaluated by an antibody inhibition procedure, i.e., sera were mixed with M. hyopneumoniae or M. flocculare antigen, incubated overnight and tested against M. hyopneumoniae antigen in the ELISA. Treatment with M. hyopneumoniae antigen markedly reduced or abolished seroreactivity, whereas treatment with M. flocculare antigen had little effect. These findings suggest that swine infected with M. flocculare usually do not develop sufficient levels of antibodies to detract from the specificity of the ELISA used to diagnose MPS.

Animals↗

Campylobacter sputorum subsp mucosalis and Campylobacter hyointestinalis infections in the intestine of gnotobiotic pigs.

At 4 days of age, 7 gnotobiotic pigs were orally inoculated with broth cultures of both Campylobacter sputorum subsp mucosalis (CSM) and Campylobacter hyointestinalis (CH). One pig was killed and evaluated each week for 7 weeks. Forty-eight hours after inoculation, CH and CSM were recovered from the feces of the pigs; thereafter, only CH was recovered. Organisms morphologically typical of Campylobacter sp were observed on the mucosal surface and on the crypt epithelial cells of the ileum, cecum, and colon from post-inoculation weeks (PIW) 2 through 7. Bacteria were clustered around the surface opening of goblet cells in pigs at PIW 6 and 7. Crypt epithelial cell proliferation and intracellular bacteria were not seen, except in 1 pig (killed at PIW 7) in which intracellular bacteria were seen only in the cecum. Therefore, CSM and CH did not induce porcine proliferative enteritis in gnotobiotic pigs.

Animals↗

Evaluation of the indirect hemagglutination assay as a practical serodiagnostic test for mycoplasmal pneumonia of swine.

Sera from swine experimentally or naturally infected with Mycoplasma hyopneumoniae (the etiological agent of mycoplasmal pneumonia of swine, MPS) were tested by the indirect hemagglutination assay (IHA), the enzyme-linked immunosorbent assay (ELISA) and the complement fixation (CF) test. The IHA detected antibody at comparable times and levels to the other 2 serological tests following experimentally-induced infection. In the late antibody response (greater than or equal to 86 days post-infection), the ELISA titres were higher than either the IHA or the CF test. The IHA appeared least satisfactory when it was used to test sera from commercial swine herds. When 1000 sera were tested, the IHA was positive for only 30 (22%) of 135 sera which were positive by the ELISA and the CF test. The IHA titres were low; 20 of the 30 sera had a titre of only 10. The end-points for the IHA were difficult to read for sera of this low titre. The relationship between positive IHA results for the herd sera obtained at necropsy, and the occurrence of gross or microscopic lesions typical of MPS was poor (41 and 50% agreement, respectively). An agreement of 39% was noted between positive IHA results and the localization of mycoplasmal antigens by an indirect immunofluorescence (IIF) test. However, IHA results correlated significantly (P less than 0.05) with gross and microscopic lesions, but not with the IIF test. No significant correlation was noted between the IHA (or the other 2 serologic tests) and the cultural isolation of M. hyopneumoniae or M. flocculare. On the basis of these results, the IHA appears to have limited promise as a practical test for the diagnosis of MPS in commercial swine herds because of the low titres observed, poor correlation of the IHA and other indicators of MPS, the necessarily subjective determination of end-points, and other inherent technical limitations of the test.

Animals↗

Serological cross-reactivity of porcine reference antisera to Mycoplasma hyopneumoniae, M. flocculare, M. hyorhinis and M. hyosynoviae indicated by the enzyme-linked immunosorbent assay, complement fixation and indirect hemagglutination tests.

The enzyme-linked immunosorbent assay (ELISA) indicated significant cross-reactivity between the antigens of Mycoplasma hyopneumoniae ( HyoP ) and M. flocculare (Floc), another porcine mycoplasma of wide distribution but uncertain pathogenic significance, when porcine antisera of each specificity were tested against HyoP antigen. The titers of the anti-Floc sera ranged from threefold to 13-fold less than the titer of the anti- HyoP reference serum at different times after immunization. These values ranged from onefold less than to fourfold greater than the minimal positive titer of 80. The antisera to the other porcine mycoplasmal antigens [i.e. M. hyorhinis ( HyoR ) and M. hyosynoviae ( HyoS )] reacted less strongly to HyoP antigen but titers only slightly less than to slightly greater than the minimal positive titer were noted for some sera. Cross-reactivity was also detected by the complement fixation test, although the titers for this test were generally lower than for the ELISA, presumably reflecting lower sensitivity of the complement fixation test. Positive indirect hemagglutination titers to HyoP antigen were also observed for both anti-Floc sera obtained at one or more times during the immune response. With two exceptions (one anti- HyoR serum with a complement fixation titer of 16 and one anti- HyoR serum with an indirect hemagglutination titer of 10), none of the anti- HyoR or anti- HyoS sera had detectable indirect hemagglutination or complement fixation titers to HyoP antigen at any time after immunization. The levels of cross-reactivity detected by the complement fixation test and indirect hemagglutination and, especially, the ELISA would be of significance for the development of any practical sero-diagnostic test for mycoplasmal pneumonia of swine.

Animals↗

Evaluation of criteria for the postmortem diagnosis of mycoplasmal pneumonia of swine.

Ten swine from each of five herds believed to be affected with mycoplasmal pneumonia of swine and ten swine from each of five herds believed to be mycoplasmal pneumonia-free were selected for postmortem study. Lungs from the 100 swine were examined; grossly and microscopically for lesions typical of mycoplasmal pneumonia of swine and culturally and by an indirect immunofluorescent procedure for the presence of Mycoplasma hyopneumoniae. Nineteen of the lungs had both gross and microscopic lesions typical of mycoplasmal pneumonia of swine and 13 (68%) of these were infected, i.e. were culturally and/or indirect immunofluorescent positive. Absence of gross lesions did not prove freedom from mycoplasmal pneumonia, 14 of 73 (19%) grossly normal lungs were found to be infected with M. hyopneumoniae. Comparison of the indirect immunofluorescent and cultural examination, as methods of diagnosing mycoplasma pneumonia, revealed that neither procedure alone was reliable in the case of negative results. Ten lungs were indirect immunofluorescent negative and culturally positive and seven were culturally negative and indirect immunofluorescent positive (11 lungs were positive by both procedures). It was concluded that a definitive diagnosis of mycoplasmal pneumonia of swine requires that M. hyopneumoniae be visualized in indirect immunofluorescent stained lung sections or that it be recovered culturally.

Animals↗

Comparison of manually performed Microtiter plate and semiautomated (cuvette) indirect enzyme-linked immunosorbent assays for the serodiagnosis of mycoplasmal pneumonia of swine.

Standardized enzyme-linked immunosorbent assays for the serodiagnosis of mycoplasmal pneumonia of swine were developed, using the traditional Microtiter plate method and a method based on semiautomated processor-analyzer instrumentation. The results of the 2 procedures were not significantly different when the titers of positive and negative anti-Mycoplasma hyopneumoniae standard reference sera were determined in triplicate in 3 separate experiments. Although both methods yielded reproducible results, the instrument-assisted enzyme-linked immunosorbent assay would be preferable if large numbers of samples are to be tested.

Animals↗

Comparison of the enzyme-linked immunosorbent assay and the indirect hemagglutination and complement fixation tests for detecting antibodies to Mycoplasma hyopneumoniae.

Caesarean-derived, colostrum-deprived swine were exposed to a broth culture of a low passage field isolate of Mycoplasma hyopneumoniae by intranasal inoculation. The intranasal-inoculated swine subsequently were commingled with their litter-mates to effect transmission via contact-exposure. Sera were collected from the swine at two to four week intervals for approximately one year postexposure and evaluated by the enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination and complement fixation tests. The intranasal-exposed swine seroconverted earlier, developed higher titers and remained indirect hemagglutination and complement fixation positive longer than the contact-exposed swine. It was concluded that the antibody response of intranasal-exposed swine was artificially high and that sera from such swine were not suitable for evaluating the sensitivity of mycoplasmal pneumonia of swine serodiagnostic tests. The indirect hemagglutination test was relatively insensitive and technically cumbersome and the least promising as a practical field test. The complement fixation test appeared to be slightly more sensitive in detecting early antibody production (especially in contact-exposed swine) but it was the least sensitive in detecting late antibodies. The ELISA was generally the most sensitive procedure. Individual high ELISA titers were from ten to 32 times greater than maximum complement fixation and indirect hemagglutination titers. The most striking difference among the three tests was the persistence of high ELISA titers late in the study. All swine were ELISA positive at necropsy approximately one year postexposure despite the fact that lungs were devoid of lesions and culturally and immunofluorescent negative for M. hyopneumoniae.

Animals↗

A microtitration agglutination test for detecting group E streptococcus infection in swine.

A microtitration agglutination test was developed and evaluated for detecting infection of swine with group E streptococci type IV, the most common causative agent of streptococcic lymphadenitis of swine. Whole cell agglutinogens representing group and type antigens of group E streptococci were tested in the microtitration agglutination test against reference antisera to Streptococcus groups A, B, C, D, E, F, G. H, K, L, M, N, O, P, Q, R, S and U, as well as specific antisera to types II, IV and V of group E. Group E specific agglutinogens were unsatisfactory in the microtitration agglutination test because of cross reactions with group P and U antisera and because of poor reproducibility of the test. Type specific agglutinogens of group E streptococci reacted only with their respective homologous antisera and not with any heterologous group antisera. None of the group E streptococci agglutinogens reacted with 52 normal swine sera. Agglutinogen made from group E streptococci type IV was selected for further evaluation in the microtitration agglutination test because group E streptococci types II and V are considered to be of minor importance in the etiology of streptococcic lymphadenitis of swine. Swine experimentally infected with a type IV strain developed significant titers in the microtitration agglutination test. All swine tested negative before exposure and seroconverted (titer >/=4) two to six weeks postexposure.The microtitration agglutination test was used by two different laboratories to test 187 duplicate samples of serum from infected swine. A total of 94.1% of the tests were read at either the same titer (48.1%) or a difference of not more than one dilution (46.0%) at the two laboratories. There was disagreement between the two laboratories in the test-positive test-negative status of 19 of the sera (10.2%). Titers of two of the sera differed by two dilutions (<4 at one laboratory and 8 at the other). The remaining 17 sera differed in titer by only one dilution (<4 at one laboratory and 4 at the other).

Agglutination Tests↗

Isolation of Mycoplasma flocculare from swine in the United States.

During 1977 to 1980, Mycoplasma flocculare was isolated from the lungs of 7 swine in the United States. Two of the swine were from a single Alabama herd, 1 was from Minnesota, 2 were from different Illinois herds, and 2 were from different Indiana herds. Three of the swine were from specific-pathogen-free herds and 4 were from conventional herds. The lungs from the 7 swine had gross lesions typical of mycoplasmal pneumonia. Two lungs yielded a mixture of M flocculare and M hyopneumoniae; the remaining 5 yielded M flocculare alone.

Animals↗

The significance of the serotype in the clinical and pathological features of naturally occurring porcine salmonellosis.

Salmonellosis was the diagnosis in 63 of 327 consecutive porcine necropsy accessions during a one year period in Indiana, an incidence four to ten times that previously reported in other areas. A herd outbreak was usually either a septicemic disease characterized by sudden mortality. Little overlap between these two syndromes were seen. Animals dying with the septicemic form usually yielded Salmonella choleraesuis var. kunzendorf on culture. Salmonella typhimurium was the usual isolate from cases of enteric salmonellosis. Oral inoculation of SPF pigs with S. typhimurium resulted in a disease similar to naturally occurring enteric salmonellosis. The pathology of porcine salmonellosis due to S. typhimurium is described.

Acute Disease↗

An antiphagocytic factor associated with group E streptococcus.

Group E streptococci (GES) grown in Tryptose phosphate broth (TPB) were highly susceptible to nonspecific phagocytosis by porcine leukocytes, whereas GES grown in serum-supplemented TPB (STPB) were resistant to nonspecific phagocytosis. It was concluded that serum supplementation induced the elaboration of an antiphagocytic factor (APF). The APF was distinct from the group-specific and type-specific polysaccharide antigens, and it was destroyed by trypsin. Electron photomicrographs revealed that GES grown in STPB possessed a fimbriated surface component similar to M protein-positive stains of group A streptococcus. The surface component was absent when GES were propagated in unfortified TPB. The APF was neutralized specifically by sera from swine immune to streptococcic lymphadenitis (SLS), a disease caused by GES. It was hypothesized that the APF was associated with the pathogenicity of GES and that it was involved in the induction of protective immunity to SLS.

Animals↗

Humoral antibody responses of swine infected experimentally with group E streptococcus. I. Whole cell agglutinin response.

Reference streptococcal antisera and sera collected from swine infected experimentally (by intranasal inoculation or contact exposure) with group E Streptococcus (GES) were studied in a tube agglutination system using whole GES cells. Specificity studies revealed common group specific antigen among GES serotypes I and III, GES strains devoid of type specific antigen (untypable by ring precipitin testing) and group P and group U Streptococcus. The group specific antigens were not agglutinated by GES type specific antisera or by group specific antisera against Streptococcus groups A, B, C, D, F, G, H, K, L, M, N, or O. Results of the study suggested that GES serotypes I and III are invalid; i.e., they are devoid of type specific antigen. Groug E Streptococcus type specific antigens II, IV, and V were agglutinated significantly only by their homologous antisera. Experimentally infected swine developed significant titers against both the group and type specific antigen of GES. Antibodies appeared from three to eight weeks postexposure and persisted for the duration of the experiment (six months). The potential utilization of the whole cell agglutination (WCA) test for detection of GES carrier swine is discussed.

Agglutination Tests↗

Humoral antibody responses of swine infected experimentally with group E streptococcus. II. Antistreptokinase and antistreptodornase responses.

Swine infected experimentally with group E Streptococcus (GES) produced significant antibody titers against streptokinase (streptococcal fibrinolysin, SK) and streptodornase (streptococcal deoxyribonuclease, SD). The antibodies directed against SK (antistreptokinase, ASK) appeared two to nine weeks postexposure and persisted for the duration of the experiment (nearly six months). The ASK inhibited SK produced by GES antigenic types III, IV, and V, by GES devoid of type specific antigen, and by a group P Streptococcus. Selected strains of GES serotypes I and II and group U Streptococcus did not produce detectable SK. The antibodies directed against SD (antistreptodornase, ASD) appeared two to three weeks postexposure, reached a peak about six weeks postexposure, and persisted at high levels for nearly six months (the duration of the experiment). The ASD inhibited SD produced by all known antigenic types of GES, by GES devoid of type specific antigen, and by strains of groups P and U Streptococcus. The antibodies failed to inhibit SD produced by group C Streptococcus. The potential utilization of ASK and ASD titers as serological means of identifying swine infected with GES (carrier swine) is discussed.

Animals↗