Uterine response to multiple inoculations with Arcanobacterium pyogenes and Escherichia coli in nulliparous ewes.

PROBLEM: Uterine infections seem more severe in nulliparous animals. Our objective was to determine whether intrauterine inoculation of nulliparous ewes with Arcanobacterium pyogenes and Escherichia coli would produce an antibody response and reduce the severity of subsequent infections. METHOD OF STUDY: Nulliparous ewes (n = 9/treatment) received (i) 'primary intrauterine inoculation' with phosphate-buffered saline (PBS) and 'secondary intrauterine inoculation' with PBS; (ii) primary PBS-secondary 75 x 10(7) cfu of A. pyogenes and 35 x 10(7) cfu of E. coli (PBS-Bacteria); (iii) primary bacteria-secondary PBS; or (iv) primary bacteria-secondary bacteria (Bacteria-Bacteria). RESULTS: Inoculations evoked an antibody response. Postmortem examinations 6 days after the secondary inoculation indicated that PBS-treated ewes did not develop uterine infections, but all bacteria-treated ewes did. Infections were either less severe or closer to resolution in Bacteria-Bacteria than they were in PBS-Bacteria ewes. CONCLUSIONS: Intrauterine inoculation of nulliparous ewes with A. pyogenes and E. coli evokes an antibody response that may help the uterus reduce the severity of subsequent infections.

Evaluation of the pathology, pathogenesis and aetiology of auricular elephantiasis in slaughter pigs.

Ears from slaughter pigs with auricular elephantiasis (n = 24) and the corresponding lymph nodes (lnn.) (n = 26) were grossly, histopathologically and microbiologically examined. Immunostaining for IgM, IgG, Cd3epsilon and bacterial antigens of Arcanobacterium pyogenes and Staphylococcus aureus was performed by indirect enzyme-based techniques. Ears were variably thickened depending on the sampled area (basis, centre and apex). However, at all locations the thickness, the length from basis to apex and the weigh of whole ears with elephantiasis were significantly increased (P < 0.01). The corresponding lnn., that is, ln. parotideus superficialis and profundus, had also increased significantly (P < 0.01) in volume. Histopathologically, lesions of the ears and the corresponding lnn. revealed changes characterized by diffuse fibrosis intermingled with multiple pyogranulomatous foci containing asteroid bodies. In the majority of lesions, four distinct zones due to different cellular infiltrates encircled the central core of the asteroid bodies. In several lesions, the pyogranulomatous foci were contained within the lymph vessels. Immunohistochemically, only the bacterial antigen of S. aureus was detected within the cytoplasm of the macrophages and/or in the asteroid bodies of the ears (41.5%) and in the regional lnn. (30.8%). An abundant number of IgM, IgG and CD3epsilon-positive cells were present in all the pyogranulomatous lesions, whereas a positive IgG-staining was observed only in a single asteroid body. Thus, porcine auricular elephantiasis is a chronic pyogranulomatous inflammation that is frequently positive for S. aureus and is lymphogenically spread. Therefore, the lesions of the ears with auricular elephantiasis and the corresponding lnn. should be termed auricular botryomycosis and botryomycotic lymphadenitis, respectively. Moreover, as the disease is observed frequently in slaughter pigs it must also be considered according to the welfare of the animals and in relation to post-mortem meat inspection.

[Neutrophil defense against bovine uterine infections]. / Neutrophile Abwehr gegen bovine Uterusinfektionen.

The killing ability of bovine peripheral blood neutrophils did not vary during the oestrous cycle. Administration of sex steroids to ovariectomised cows reduced the intrinsic killing ability of neutrophils but enhanced the opsonising ability of serum. Exudate from experimental uterine infection with Corynebacterium pyogenes and Fusobacterium necrophorum impaired neutrophil function, probably as a result of the action of bacterial leucotoxins.

Effect of infection of the genital tract on the concentration of IgG and albumin in bull serum and semen.

Bovine IgG and albumin concentrations were determined from serum and semen of 59 bulls that were divided into 4 groups: 35 non-infected bulls (Group 1); 10 with vesiculitis due to Corynebacterium pyogenes (Group 2); 10 bulls with orchitis due to Chlamydia psittaci (Group 3); and 4 bulls with infectious vesiculitis (Group 4) sampled both before and after antibiotic treatment. Serum IgG concentrations (25 mg/ml approximately) were similar in non-infected (Gp 1) and infected bulls (Gp 2,3,4) whereas serum albumin concentrations were greater in infected than in non-infected bulls (51 mg/ml vs.41 mg/ml; p less than 0.01). By contrast, both semen IgG and albumin concentrations in infected bulls (0.47 and 0.54 mg/ml respectively) were significantly different from those of non-infected bulls (0.14 and 0.32 mg/ml; p less than 0.01). In addition, bulls with chlamydial orchitis had both semen (but not serum) IgG and albumin levels higher than those suffering from vesiculitis (p less than 0.01). Antibiotic therapy led to recovery and simultaneously to decreased concentrations of semen IgG and albumin. These results strongly suggest a local IgG synthesis or selective diffusion after such genital infections and further indicate that semen IgG and albumin assays could be a new and valuable tool for diagnosis and evaluation of genital infections.

Identification of bovine lymphocyte subpopulations by a combined bacterial adherence and fluorescent antibody technique.

It is known that certain strains of bacteria bind selectively to subpopulations of human peripheral blood lymphocytes. We have developed a technique which used the specificity of bacterial binding concurrently with fluorescent antibody staining methods to identify 5 B-cell and 5 T-cell subpopulations of bovine lymphocytes. In addition, greater than 95% of the peripheral blood lymphocytes could be positively identified as being either T-cells or B cells. Using ethidium bromide-stained bacteria and lymphocytes in combination with fluorescent antibody staining to detect surface immunoglobulins or T-cell antigens, the method provided a simple yet highly specific technique for the enumeration of both B and T cells in 1 preparation of peripheral blood lymphocytes. The use of bacterial rosetting with fluorescent antibody staining was found to be easier and more reliable than the methods currently used to identify bovine B- and T-lymphocyte subpopulations.

Cross-reactions between mycobacteria. II. Crossed immunoelectrophoretic analysis of soluble antigens of BCG and comparison with other mycobacteria.

Cross-reactions between Mycobacterium bovis BCG and various other mycobacteria, Nocardia asteroides, Corynebacterium pyogenes and Listeria monocytogenes were studied by incorporating antibodies against these bacteria in the intermediate gel of a crossed immunoelectrophoretic system with BCG antigen and anti-BCG antibodies. In the BCG reference system forty-four distinct antigenic components were recorded, of which thiryt-three cross-reacted with Mycobacterium tuberculosis, twenty-five with M. avium, twenty-one with M. suvalii, eighteen with M. smegmatis, Fifteen with M. nonchromogenicum, twelve with M. phlei, eight with N. anteroides and two with C. pyogenes, whereas no cross-reaction was detected with L. monocytogenes. The value of the method for characterization of mycobacterial antigens is discussed. A taxonomic system based on this method appears particularly valuable for studies of non-cultivativable mycobacteria such as M. leprae. A majority of twenty-one patients with lepromatous leprosy had anti-BCG antibodies of restricted specificity, affecting only four or five BCG antigens, although one patient had twelve anti-BCG specificities. Most of these antibodies reacted with those BCG antigens that cross-react extensively with other mycobacteria.

[Immunofluorescence tests for the detection of antibodies against Corynebacterium pyogenes and streptococci in blood serum and vaginal mucus of cattle]. / Immunofluoreszenzuntersuchungen zum Nachweis von Antikörpern gegen Corynebacterium pyogenes und Streptokokken im Blutserum und Vaginalschleim von Rindern.

Bacteriological tests were applied to cattle with endometritis and vaginitis. Included were cervical mucus samples and immunofluorescence tests to detect in that mucus as well as in blood serum antibody to Corynebacterium pyogenes and Streptococcus haemolyticus. The results pointed at intensive contact of the animals with the above pathogens and to their frequent occurrence in cervical mucus of cattle afflicted with endometritis and vaginitis. They also supported the assumption of localised antibody formation in the sexual organs or female cattle.

Precipitating antibody against protease of Corynebacterium pyogenes in pigs.

Antibody in sera from pigs carrying an abscess associated with Corynebacterium pyogenes and healthy pigs was examined by the agar gel diffusion test. In the test, the concentrated culture fluid containing the protease of C. pyogenes was used as antigen. As a result, precipitating antibody was demonstrated in sera from 25 of 30 abscessed pigs and a few of the healthy pigs. When the relationship between precipitating antibody and protease was examined by the immunoelectrophoresis and gel filtration of the concentrated culture fluid, the antibody was shown in the same position as the protease. From the result, it was clear that the precipitating antibody was against the protease of C. pyogenes. All the proteases produced by 27 strains of C. pyogenes of porcine and bovine origin were serologically identical with one another. They were, however, serologically different from those of Staphylococcus aureus, Bacillus cereus, and B. subtilis. In the inhibition test, the proteolytic activity of C. pyogenes was inhibited by the serum of the abscessed pig. It was also inhibited by healthy pig serum. From the results, it seems that the determination of precipitating antibody may be useful for the diagnosis of C. pyogenes infection.

Antibody response to and immunity induced by Corynebacterium pyogenes vaccine.

A procedure is described for producing a high quality C. pyogenes vaccine. A satisfactory antibody response was obtained by administering 3 subcutaneous injections of vaccine at 10-day intervals, using doses of 2,0 ml for rabbits, 5,0 ml for sheep and 10,0 ml for cattle. The titres resulting from this procedure compared favourably with those obtained from following the more intensive schedule and higher doses of vaccine recommended by the British Veterinary Codex. The antibody response was at its best 1-2 months after immunization but dropped to a plateau 6 months after immunization. Mice were also successfully immunized against challenge with 1,5x108 organisms by the application of 3 doses of vaccine administered at 10-day intervals.

Corynebacterium pyogenes and bovine abortion.

In the examination of bovine fetal material it was found that there was a significant increase in the proportion of mixed infections identified as the time between abortion and the collection of the samples increased. Examination of paired serum samples from abortions from which only Corynebacterium pyogenes was isolated revealed evidence of active infection in two-thirds, suggesting that C. pyogenes may have been acting as a primary abortifacient in these cases.