Germicidal activities of representatives of five different teat dip classes against three bovine mycoplasma species using a modified excised teat model.

Six representative teat dips from five different teat dip classes were tested for germicidal activity against challenge exposure to Mycoplasma bovis, Mycoplasma californicum, and Mycoplasma bovigenitalium using a modified excised teat model. All teat dip formulations tested were efficacious against all of the Mycoplasma species, providing bacterial logarithmic reductions above 4. The germicides performed best against M. bovigenitalium with an average log reduction (LR) of 6.29. Average LR were 5.41 and 5.70 against M. bovis and M. californicum, respectively. The iodine and chlorhexidine products performed best against M. bovis and M. californicum, respectively, with complete kill of all organisms. The chlorhexidine and the barrier chlorine product also had complete kill of M. bovigenitalium organisms.

The effect of chronic immunostimulation of the nonlactating bovine mammary gland with interleukin-2, pokeweed mitogen, and lipopolysaccharide.

Interleukin-2, pokeweed mitogen, and lipopolysaccharide were evaluated for their ability to accelerate involution and to stimulate local cellular defenses in the nonlactating bovine mammary gland. Twelve cows were divided into three treatment groups of 4 cows each to receive interleukin-2, pokeweed mitogen, or lipopolysaccharide. One day after drying off, 3 mammary quarters of each cow were infused with 100 micrograms of immunostimulant daily for 21 d; the remaining control quarter received PBS. Secretion samples were collected weekly to determine bacteriologic status, total SCC, and differential cell counts. On d 21, cows were killed, and tissues were collected for microscopy. Overall, SCC were higher in immunostimulated quarters, but only those infused with interleukin-2 were significantly elevated over controls. By wk 3, the percentage of neutrophils decreased in interleukin-2 and pokeweed mitogen quarters over pretreatment values, percentage of macrophages increased in interleukin-2 quarters, and percentage of lymphocytes increased in pokeweed mitogen and lipopolysaccharide quarters. Percentage of alveolar lumina was reduced, and connective tissue stroma increased, in all immunostimulated quarters compared with those of controls, suggesting accelerated involution. Involution was greatest in quarters treated with interleukin-2. Leukocyte infiltration was greater in immunostimulated quarters than in control quarters. Similarly, concentrations of Ig-producing plasma cells were greater in immunostimulated quarters than in control quarters. Quarters infused with interleukin-2 exhibited the greatest concentration of plasma cells, followed by quarters treated with pokeweed mitogen and lipopolysaccharide; IgG1 plasma cells predominated, followed by IgG2, IgA, and IgM. Interleukin-2 accelerated involution and stimulated local antibody production more than did the two mitogens, suggesting a potential role for this cytokine as a general immunostimulant at drying off.

Histological response of the bovine mammary gland to intramammary devices.

Histologic response of the bovine mammary gland to presence of three intramammary device models (abraded, star, or grooved) was studied in 12 lactating cows. Uninfected quarters fitted with devices exhibited greater leukocyte infiltration into teat and gland cistem linings as well as into mammary parenchyma adjacent to the gland cistem compared with unfitted control quarters. Cytologic alterations to cistem linings in device-fitted quarters ranged from degeneration and sloughing of surface cells of the double-layered epithelium to hyperplasia, stratification, and keratin formation. In uninfected quarters, quantification of mammary parenchymal components showed no differences among models for percentages of epithelium, but percentage of lumen was lowest and stroma highest for the star intramammary device model, suggesting reduced secretory activity or mammary involution in response to these intramammary devices. Presence of bacterial infection amplified the histologic responses to all devices. Leukocyte infiltration remained greater in device-fitted quarters compared with controls but was elevated over uninfected quarters for all treatments. Likewise, hyperplasia, stratification, and keratin formation of the cistem epithelial lining were more frequently observed in infected quarters. Among models in infected quarters, percentage of lumen was lowest and stroma highest in quarters fitted with abraded devices. In most cases, presence of infection masked any effect of devices on mammary parenchymal components. Plaque formation was observed on all models and tended to be thicker on those retrieved from infected quarters. Electron microscopic examination showed that plaque was composed of leukocytes, cell debris, keratin, and amorphous material. Results demonstrated that most intramammary device models were successful in stimulating leukocytosis into the gland, and tissues from fitted quarters displayed alterations to cisternal linings; however, quarters fitted with these devices exhibited reduced secretory activity.

Histological response of bovine mammary tissue to an intracisternal bead device.

Placement of an intracisternal bead device into teat cisterns of dairy cows at drying off was studied. Histological analysis of teat cisternal tissues from quarters fitted with devices demonstrated significant elevation in leukocyte infiltration and hypertrophy of the epithelial lining. Tissue changes ranged from degeneration and sloughing of surface cells of double-layered epithelia to marked thickening and lysis of cisternal epithelium, leaving only the basement membrane intact. Quantification of mammary parenchymal components from quarters fitted with devices demonstrated no differences in percentage area occupied by alveolar epithelium, lumen, and intraalveolar stroma from unfitted control quarters. However, quantification of leukocyte numbers demonstrated slightly greater leukocytosis into tissues of quarters fitted with devices in comparison with controls. Devices were successful in stimulating a localized leukocytosis into teat cisternal tissues, which may serve to amplify the leukocyte response to mastitis-causing organisms once they penetrate the teat duct.

Prevalence and ultrastructural characteristics of bovine mammary corpora amylacea during the lactation cycle.

Morphologic and histochemical study of spherical, lamellated inclusion bodies in bovine mammary parenchyma established presence of corpora amylacea. The majority (90%) were in alveolar lumina, followed by stroma (7%) and epithelium (3%). Occurrence of corpora amylacea was unrelated to lactation age, intramammary infection status, milk somatic cell concentration, or milk production. Quantitative analysis demonstrated a progressive increase of prevalence of these structures from the early to later stages of lactation, followed by a return during the late dry period to concentrations at early lactation. Ultrastructural study revealed deeply basophilic, dense, lamellated deposits (70%) and less dense, fibrillar deposits (30%). Dense bodies stained partially or not at all for amyloid but stained for presence of calcium salts, whereas fibrillar bodies were amyloid positive and calcium negative. Corpora amylacea were most prevalent in fully differentiated parenchymal areas (47.1 bodies/unit tissue area of 2.4 X 10(6) micron 2), less numerous in intermediately differentiated areas (23.4), and sparse in nonsecretory areas (7.1). Morphologic relationships between deposits and parenchyma suggest that corpora amylacea suppress milk secretion and flow during late lactation in isolated areas by engorging luminal spaces and clogging small ducts, leading to milk stasis and involution.

Evaluation of protein A and a commercial bacterin as vaccines against Staphylococcus aureus mastitis by experimental challenge.

Protein A and a commercial staphylococcal bacterin were evaluated by experimental challenge with Staphylococcus aureus (ATCC 29740). Thirty cows in first lactation were in three treatment groups, protein A, bacterin, and nonvaccinated controls. Studies were through three lactations and included bacteriological and cytological analyses of quarter milk samples. Rate of intramammary infection with Staphylococcus aureus was similar for vaccinated and unvaccinated cows. Rates of spontaneous cure within each lactation were significantly higher for vaccinated cows. For all three lactations, spontaneous cure rates were 83, 73, and 47% for protein A, bacterin, and control cows. Somatic cell counts were significantly lower for vaccinated cows for quarters infected with Staphylococcus aureus, but no differences were demonstrated for milk production by lactation. Incidence of clinical mastitis was higher in unvaccinated cows, but too few developed for a valid comparison.

Distribution, location, and ultrastructure of plasma cells in the uninfected, lactating bovine mammary gland.

Quantitative cytological analysis demonstrated a marked and progressive increase in concentration of subepithelial plasma cells from milk-secreting parenchyma to the distal teat end mucosa in the normal, lactating bovine mammary gland. Parenchymal plasma cells exhibited typical ultrastructure with abundant, flattened stacks of rough endoplasmic reticulum, and hypertrophied Golgi components, while many cells in the teat end mucosa displayed an ergastoplasm distended with fine, flocculent material. Intraepithelial plasma cells were also observed in teat end mucosa. Results suggest that teat end tissues function by recruiting plasma cells to serve as protection against invading mastitis-causing organisms.