Affinity of microorganisms of the genus Ureaplasma to the reproductive organs of cattle

The purpose of this work was to define more precisely the role of Ureaplasma organisms in the aetiology of granular vulvovaginitis and balanoposthitis (GVVBP) of cattle. To contribute to this question the frequency and degree of infection with Ureaplasmas in two main groups of cattle was taken into account: (a) in cattle with symptoms of the mentioned disease, (b) in cattle without clinical symptoms. The samples of semen from 301 sires with symptoms of GVVBP and from 43 healthy sires as also vaginal mucus swabs from 96 cows with GVVBP and from 40 cows mated by the sire infected with Ureaplasma organisms and from 50 cows inseminated with semen which contained Ureaplasma organisms were taken for bacteriological examinations. The control group in relation to the above mentioned cows constituted of 22 heifers free from symptoms of GVVBP and neither inseminated nor mated naturally. It has been shown that on an average 78.1% of sires with pathological changes in the mucosa of the penis or prepuce and only 25.6% of healthy sires were infected with Ureaplasma organisms. The concentration of Ureaplasma organisms was also significantly higher in material obtained from sires with symptoms of the disease than in that from healthy animals. Ureaplasma organisms were demonstrated more frequently (72.7%) in cows with GVVBP than in cows without these symptoms (13.3%). Similarly, as in the material obtained from sires, in the material taken from cows with symptoms of the disease the concentration of Ureaplasma organisms was significantly higher than that in the material originating from the healthy cows. The obtained findings may indicate that Ureaplasma organisms play a role in the aetiology of GVVBP.     

Infectious bovine keratoconjunctivitis: Bacteriologic,immunologic, and clinical responses of cattle to experimental exposure with Moraxella bovis

The bacteriologic, immunologic, and clinical responses of 3- to 4-month old Holstein-Friesian calves to experimental exposure with Moraxella bovis type 10900 has been investigated. After u.v. radiation and intraconjunctival exposure with 1.9 × 10⁷ microorganisms, each eye of 16 calves exhibited signs of blepharospasm, photophobia, and increased lacrimation. Bacteria were recovered from exposed eyes for 2–7 consecutive weeks before maximal clinical response occurred. The severity of the cases varied from eyes that exhibited mild signs to severe clinical cases with profuse lacrimation, conjunctival swelling, corneal opacity, and ulceration. By 70 days after exposure, M. bovis could not be recovered from any conjunctival swabs, and clinical signs were not observed. Four non-exposed control animals did not develop clinical signs nor was M. bovis recovered from conjunctival swabs.Lacrimal secretions collected at the time of and 1 week after maximal clinical response had significantly elevated levels of total protein as compared to those collected 3, 2, and 1 week before, and 2 and 3 weeks after maximal clinical response. A passive hemagglutination test, using tanned formalized sheep erythrocytes sensitized with M. bovis sonicate antigen, detected antibody in lacrimal secretions from 22 of 32 eyes. The appearance of specific antibody in lacrimal secretions correlated with the amelioration of clinical signs and the decline in numbers of M. bovis microorganisms recovered from conjunctival swabs.     

Potential use of lymphocyte blastogenic responses in diagnosis of bovine tuberculosis

A comparison of in vitro lymphocyte responses and delayed type tuberculin skin test responses was made in an animal experimentally exposed to a Mycobacterium bovis-infected animal and in cattle naturally infected with M. bovis. Tuberculin skin tests did not suppress in vitro lymphocyte responses to M. bovis PPD and to M. avium PPD tuberculin. The whole blood test used in these studies provided for considerable savings in time as compared to use of purified lymphocytes for evaluating in vitro cellular responses. Variations in the responsiveness of lymphocytes to specific mycobacterial antigens was observed, therefore, it is recommended that profiles be established using three or more tests conducted at 14-day intervals.     

Differential responses of cells from human skin keratinocyte and bovine mammary epithelium to attack by pore-forming Staphylococcus aureus alpha-toxin

Human skin keratinocytes HaCat attacked by Staphylococcus aureus α-toxin showed a transient drop of cellular ATP levels whereas in toxin-perforated bovine mammary epithelial cells (BMEC), the ATP levels dropped more slowly. Morphologically, during the ATP level depletion, HaCat cell developed a spacious intracellular vacuole together with the transient influx of trypan blue. WST-1 signal, which tested the function of mitochondrial enzyme in viable cells, also decreased concomitantly. On the other hand, BMEC excluded trypan blue and vacuolation was not observed throughout the experiment. We conclude that mammary epithelial cells resist the toxin better than keratinocytes. This is the first report showing that α-toxin enhances transient membrane permeability to large molecules, temporary vacuole formation and the transient defect of mitochondrial enzyme in viable cells without cell lysis.     

Laboratory diagnosis of respiratory diseases of cattle

The authors describe the procedure of laboratory diagnosis for bovine respiratory diseases: direct diagnosis by isolation and for identification of bacteria or viruses and indirect diagnosis by serological methods. They specify the restraints and limits of this diagnosis and the significance results which are obtained and connected with knowledge of anamnestic information.     

A comparison of the mitogenic effects of a phorbol ester on lymphocytes from bovine spleen,lymph node and peripheral bloodComparaison des effets mitogenes d'un ester de phorbol sur des lymphocytes bovins de rate,de ganglions lymphatiques et de sang peripherique

A comparison of the mitogenic effects of a phorbol ester on lymphocytes from bovine spleen, lymph node and peripheral blood.Bovine lymphocytes from three tissues, lymph node, spleen and peripheral blood were compared for their mitogenic responses to 12-O-tetraecanoylphorbol-13-acetate (TPA), a phorbol ester tumor promoter. TPA alone was found to be either not mitogenic or caused only a weak response when compared with the mitogenic lectin phytohemagglutinin (PHA). Of the three lymphocyte preparations, blood cells showed the greatest proliferative response to TPA. However, all three, lymph node, blood and spleen cells, showed a co-mitogenic response to TPA. That is, TPA synergistically enhanced DNA synthesis in cells stimulated with a suboptimal concentration of PHA.     

Bovine adenoviruses—IV. Two mixed antigens for routine serodiagnosis by complement fixation reaction

The frequency of bovine adenovirus infections occurring in cattle has been grossly underestimated up to the present day. Primary isolation of serotypes belonging to bovine subgroup II is cumbersome. Hardly any laboratory has used all 8 officially-recognised bovine serotypes in seroneutralisation-tests, which should be done when this type-specific method is used for serodiagnosis. The complement fixation test, known as group-reactive from human adenovirus serology, has failed to disclose the true incidence of bovine infections, as until recently the importance of a novel additional group-reactive bovine adenovirus antigen had been undisclosed. Here we describe production, composition and performance of two mixed antigens for complement fixation reactions, which take into account recent findings by our team on peculiarities of bovine adenovirus antigens. Mixed antigen 1 contains bovine serotypes 1, 2 and 3 and detects group-specific antibodies of the classical mastadenoviruses. Mixed antigen 2 contains bovine serotypes 4, 5, 6, 7 and 8 and determines group-specific antibodies against the novel paramastadenoviruses. In addition, each antigen is capable of demonstrating complement-fixing type-specific antibodies in sera against the respective types included in each mixed antigen, with a net enhancing effect produced by mixing. Use of the 2 mixed antigens promptly shows whether bovine adenoviruses, presently recognised as well as unclassified, are involved in a given outbreak of respiratory or enteric disease of cattle. If needed, the responsible type may be determined in a second step of serological investigation.     

Non-immune erythrocyte rosette formation of bovine peripheral blood and thymus lymphocytes

An E-rosetting reaction is described which gave 92.1%±2.4 (mean±S.D.) E-rosettes with bovine fetal thymocytes and 48.2%±8.4 with with bovine peripheral blood leukocyte (PBL) preparations. Both culture conditions and culture medium were critical factors in obtaining maximal and reproducible E-rosette numbers. Optimum rosette formation occurred when bovine PBL and neuraminidase treated sheep erythrocytes (nSRBC) were reacted in L-15 culture medium supplemented with 10% fetal calf serum (FCS). Other media including 100% FCS, MEM with 10% FCS, and RPMI-1640 with 10% FCS were less satisfactory. Cultural conditions found to be optimal for enumeration of bovine E-rosettes are similar to those reported as optimal for detection of human T cells. The specificity of rosette formation by bovine thymus derived (T) lymphocytes was shown by demonstration of (1) rosettes and surface membrane immunoglobulins (mIg) on different cells in PBL, (2) rosette formation by the majority of fetal thymocytes, and (3) no inhibition of rosette formation by anti-immunoglobulin serum. Using the E-rosette and mIg assays for presumptive bovine T and B lymphocytes, respectively, it was possible to differentiate from 57.5 to 90% (75.2%±9.3) of cells in bovine PBL preparations, and from 90.2 to 97.5% (94.2%±2.1) of cells in bovine fetal thymocyte preparations into T and B cells.     

The metabolism and transport of bovine serum SIgA

Experiments were undertaken in six Holstein-Friesian cattle to determine whether secretory IgA (SIgA) could be transported from serum into exocrine body fluids. Preliminary data indicated that when IgG₁, IgG₂, IgM and SIgA were administered i.v., only SIgA and IgG₁ appeared in bile 90 min later at concentrations equal to or exceeding those in serum at the same time. Two hours post-injection, 70% of the SIgA recovered in bile was intact however only 30% co-precipitable with anti-secretory component (SC) while >90% of the administered IgA was precipitated by this method. All recovered IgG₁ was of low molecular weight. More detailed studies indicated that the IgA recovered in bile 7 h post-injection or in milk 3 h post-injection, was predominately lower molecular weight than intact SIgA. Most of this low molecular weight radioactivity was TCA precipitable and ca 50% was dialyzable; these data indicate that TCA-precipitability is an inadequate criterion for determining whether intact SIgA is transported. The radioactivity recovered in parotid saliva was almost entirely non-TCA precipitable and dialyzable. Almost all SIgA recovered in bovine serum remained intact and had a of 15.7 h. When transport into milk and bile was calculated from total, recovered radioactivity (i.e. 29% and 2.7, respectively), data compared favorably with those conducted in sheep in which dimeric IgA (without SC) was administered i.v. When we calculated transport on the basis of recovered intact IgA, only 1.47 and 0.54% of the injected dose had been transported into milk and bile, respectively, 24 h later. Most IgA in ruminant bile may be of serum origin although the same appears to be unlikely for the IgA in milk.     

Characterization and separation of bovine lymphocyte populations

Bovine lymphocyte populations were characterized by surface markers, rosette-forming ability and behaviour towards mitogens. After pre-treatment with neuraminidase 16% of the bovine blood lymphocytes and 14% of the bovine spleen cells formed spontaneous (E) rosettes with sheep erythrocytes. About 20% EAC rosette-forming cells were detected among both cell populations. Protein A receptors were detectable among 8% of the blood lymphocytes and 26% of the spleen cells. Bovine lymphocytes responded to pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (Con A). An enrichment of bovine B and T cells was obtained by E-rosette sedimentation (81–84% B cells) and by filtration through nylon fiber columns (51–65% T cells). The T cells obtained after nylon filtration still responded to the mitogens PHA, Con A and PWM. Enriched B-cell populations responded to bacterial lipopolysaccharide (LPS). After monocyte depletion the mitogenic response of blood lymphocytes was not influenced.