S.C. Nickerson

SECRETION COMPOSITION DURING BOVINE MAMMARY INVOLUTION AND THE RELATIONSHIP WITH MASTITIS

1. Bacteriological analysis revealed that 30% of quarters contained coagulase-negative staphylococci, Staphylococcus aureus, Corynebacterium bovis, or streptococci. 2. As involution progressed, somatic cell counts, percent protein, pH, and concentrations of serum albumin, lactoferrin, and immunoglobulin G increased while percent fat, concentrations of citrate, and the citrate to lactoferrin molar ratio decreased. 3. Mammary secretion from infected quarters had significantly higher numbers of somatic cells, percent polymorphonuclear leukocytes, and pH, but lower percentage lymphocytes, fat, and lactoferrin concentrations compared to uninfected quarters. 4. Results suggest intramammary infection altered normal secretion composition during bovine involution and lactogenesis. 5. Lower levels of antibacterial components in bovine mammary secretion during the peripartum period may have reduced the natural defense potential of the gland.

Mastitis and its Impact on Structure and Function in the Ruminant Mammary Gland

It is a given in biology that structure and function go hand-in-hand. At the level of the mammary alveoli, copious milk production depends on the proliferation of mammary epithelial cells and the biochemical and structural differentiation of these cells after parturition. For example, data from quantitative structural studies demonstrate that differences in milk production between beef and dairy cows correspond with a relative failure of alveolar cell differentiation in cattle not specifically selected for milk yield. It is likely, but not proven, that production differences within or between dairy breeds are also determined by differences in the capacity of alveolar cells to differentiate or to maintain an adequate state of differentiation. These observations strongly support the belief that insults from mastitis that lead to losses in mammary function are directly related to disruption of alveolar cell integrity, sloughing of cells, induced apoptosis, and increased appearance of poorly-differentiated cells. Ironically, reduced milk production in cases of subclinical mastitis, is also associated with increases in milk somatic cell count. Thus the elevated neutrophil migration evoked to fight inflammation can inadvertently rendered alveolar epithelial cells non-secretory. A challenge to future researchers will be to devise mastitis treatments and therapies that prevent and/or repair damage to alveolar structure and maximize subsequent secretory cell differentiation.

Control of heifer mastitis: antimicrobial treatment-an overview.

Initial studies in Louisiana, USA to determine the prevalence of mastitis in breeding age dairy heifers demonstrated that intramammary infections (IMIs) were present in 97% of heifers and 75% of quarters. Most common isolates were Staphylococcus aureus, Staphylococcus hyicus, and Staphylococcus chromogenes; somatic cell counts (SCCs) ranged from 12.4 to 17.3 x 10(6)ml(-1). Histologic examination of Staph. aureus-infected quarters demonstrated significant reductions in alveolar epithelial and luminal areas, and increases in connective tissue and leukocytosis, illustrating limited secretory development and marked inflammation. A one-time infusion of various nonlactating cow antibiotic preparations into infected quarters during different stages of gestation but >45 days prepartum resulted in cure rates for Staph. aureus IMI of 67-100%. Mean SCC was 50% lower at calving for treated heifers, and milk yield over the first 2 months of lactation was 10% greater than that of untreated controls. Subsequent multiple herd studies, however, revealed that use of nonlactating cow therapy was beneficial only in herds exhibiting a high prevalence of heifer mastitis and not in low prevalence herds. Results of lactating cow antibiotic therapy infused 1-2 weeks prepartum demonstrated cure rates of 59-76% vs. 26-31.7% in untreated controls. In some studies, milk production during the first lactation in treated heifers was approximately 10% higher than untreated controls, and SCC were significantly lower; however, in other studies, prepartum treatment was successful in reducing prevalence of infection but had no effect on SCC or milk yield during the subsequent lactation. Thus, treatment of heifers is advantageous because the cure rate is much higher than during lactation, there is no milk loss, and risk of antibiotic residues minimal; however, successful therapy may not necessarily result in lowered SCC and increased milk production in all herds.

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.

A comparison of the STAPH-ident and STAPH-Trac systems to conventional methods in the identification of staphylococci isolated from bovine udders

The STAPH-Ident and STAPH-Trac systems (Analytab Products, Plainview, N.Y.) were compared to conventional methods for identification of staphylococci isolated from bovine udders. The STAPH-Ident system identified 80.5% of isolates correctly. An additional 7.6% of Staphylococcus hyicus strains were delineated from S. epidermidis by characterizing acetoin and pigment production. Final accuracy of the STAPH-Ident system was 88.1%. The STAPH-Trac system identified 66.1% of isolates. Negative phosphatase tests for 42.3% of S. hyicus strains resulted in misidentification as S. simulans. Consequently, only 45.5% of S. hyicus isolates were identified correctly by the STAPH-Trac system. Minor modification of each system would permit accurate, rapid identification of staphylococci isolated from bovine udders.

Effect of dietary supplementation on the antimicrobial activity of blood leukocytes isolated from Holstein heifers

The purpose of this investigation was to evaluate the effect of an immunostimulating feed supplement (OmniGen-AF®) on the antimicrobial properties of blood leukocytes in dairy heifers in an attempt to prevent mastitis. Blood leukocytes from supplemented and unsupplemented controls were used. Phagocytic activity and reactive oxygen species (ROS) production were studied on d 0 (prior to feed supplementation) and on days 30 and 60 after supplementation. L-selectin and IL-8R mRNA expressions on blood leukocytes were evaluated on d 0 (prior to feed supplementation) and monthly thereafter for 15 mo. On d 30 after supplementation, neutrophils from treated heifers exhibited greater binding and internalization of Escherichia coli and greater ROS production compared with unsupplemented controls. L-selectin mRNA expression was increased in supplemented heifers vs. controls; however, IL-8R mRNA expression was not different. Results support the continued study of dietary supplementation as an additional management tool to enhance udder health in dairy heifers.

A case study of Streptococcus group G infection in a dairy herd

In this paper an outbreak of bovine mastitis in a dairy herd caused by Streptococcus group G is described. Initial identification of the organism as Streptococcus agalactiae was based upon hemolysis, esculin reaction, and CAMP reaction observed on blood agar used for bulk milk analysis. Initial therapy with a penicillin-containing, lactating cow product cured 24.4% of all streptococcal infections. Definitive serogrouping by coagglutination determined that the majority of infections were due to a weakly-hemolytic, esculin-negative Streptococcus group G. Treatment with a cephalosporin, lactating cow product was only moderately successful (54.9%). Dry-cow therapy with 300 mg cephalosporin eliminated 69.5% of refractory infections. Animals remaining infected following dry-cow therapy were culled. Histopathological study of parenchymal tissue in the lower portion of infected quarters revealed mild damage and slight involutionary changes, whereas, deep parenchymal areas appeared relatively unaffected.

Evaluation of an Anti-Inflammatory Factor Derived from Hyperimmunized Cows

Abstract An anti-inflammatory factor isolated from milk of hyperimmunized cows was analyzed in vitro and in vivo. Macrophages collected from lacteal secretions of a unimmunized nonlactating cow showed increased ability to kill phagocytosed Staphylococcus aureus when incubated with the anti-inflammatory factor. Mice injected intraperitoneally with 10 mg/kg of anti-inflammatory factor demonstrated an increased LD50 to S. aureus when challenged intraperitoneally. Injected mice also demonstrated significantly (P < 0.05) less mammary inflammation and involution and increased clearance of S. aureus when challenged intramammarily. Quantitative histologic analysis of mammary tissues from mice injected with antiinflammatory factor demonstrated significantly (P < 0.05) more lumen, less interalveolar connective tissue, and less leukocytic infiltration compared with control mice. Mammary glands of mice injected with anti-inflammatory factor and challenged with S. aureus also contained fewer colony-forming units than control mice. The product appeared to exert its effect on the nonspecific defense system via modulation of leukocyte function.

Modulation of innate immune function and phenotype in bred dairy heifers during the periparturient period induced by feeding an immunostimulant for 60 days prior to delivery

The purpose of this study was to evaluate the effect of a feed additive (OmniGen-AF(®), reported to have immune modulating activity) on innate immunity and health events during the periparturient period in dairy heifers when immunity is suppressed. From 60 days prepartum through calving, supplemented heifers (n=20) received OmniGen-AF(®) daily and were compared with unsupplemented controls (n=20). Blood leukocyte innate immune activity (phenotype markers, phagocytic activity, and reactive oxygen species--ROS production) was measured prior to feeding (60 days prepartum), 30 days later, and on days 1, 7, 14, and 30 postpartum. Adverse health events (udder edema, ketosis, displaced abomasum, and death) and milk production were measured at calving and into early lactation. The fraction of leukocytes with measurable CD62L (L-selectin) on their surface from supplemented heifers tended to be greater during the periparturient period in treated heifers than controls (p=0.100). Likewise, leukocyte phagocytosis of Escherichia coli and Staphylococcus aureus during this time period tended to be greater in heifers supplemented with OmniGen-AF(®) (p=0.100). Conversely, ROS production in response to phorbol myristate acetate or when leukocytes were stimulated with killed S. aureus lysate tended to be greater among control heifers compared with supplemented animals (p=0.100). Supplemented heifers exhibited fewer incidents of udder edema than controls (p=0.030) and tended to exhibit a lower rate of new cases of mastitis (p=0.098); however, no differences were observed in milk somatic cell counts or level of milk production. Results demonstrate a positive role of OmniGen-AF(®) in amplifying leukocyte function consistent with antibacterial activity during the periparturient period, and support the continued study of dietary supplementation to enhance mammary gland health in dairy cows.

Cilia on bovine mammary epithelium: ultrastructural observations.

SummaryUltrastructural examination of bovine mammary tissues revealed the presence of 9 + 0 or primary cilia protruding from surfaces of alveolar epithelial and myoepithelial cells. Cilia of epithelial cells protruded approximately 1200 nm into lumina of alveoli and arose from a basal body centriole, the associated centriole of the diplosome, and an accessory rootlet system. Cilia on epithelial cells were more frequently observed than cilia on myoepithelial cells. Occasional cilia made contact with macrophages in the alveolar lumen. The structures were more commonly found in tissues from nonlactating cows, and most were observed in the ventral portion of the mammary gland.