Supramammary Lymph Nodes Research Articles

Immunophysiological activity of supramammary lymph nodes of the ewe during the very early phase of staphylococcal mastitis

Efferent mammary lymph was collected from lactating ewes which were unimmunised (controls) or immunised during pregnancy with two doses of an attenuated live Staphylococcus aureus vaccine either in the hindlimb ("directly primed' supramammary nodes) or in the brisket ("indirectly primed' supramammary nodes). Mammary lymph was also collected from unimmunised animals in which the supramammary nodes had been extirpated several months before. Ewes in which the supramammary nodes had been directly primed by staphylococcal vaccination before challenge had a significantly greater output of IgM- and IgG2-containing cells in lymph and higher concentrations of IgG1 and IgG2 antibody against S aureus surface antigens than did other groups. Lymphadenectomised ewes had fewer leucocytes in mammary lymph but a much higher proportion of neutrophils than other ewes, indicating that afferent mammary lymph has an unusually high number of neutrophils and most of these cells are filtered out in the supramammary lymph nodes under normal circumstances. The results indicated that most of the leucocytes in efferent lymph were derived from the supramammary nodes. After induction of experimental staphylococcal mastitis there was a rapid drop in leucocyte output in lymph within one to four hours after infection; the data indicated that events within the supramammary nodes were responsible for this phenomenon. The output of immunoglobulin-containing cells was reduced during this phase. No significant increases in output of lymphoblasts, immunoglobulin-containing cells or specific antibody occurred during the six hours immediately following infection.

Isolation of Brucella abortus from supramammary lymph nodes of cattle from infected herds vaccinated with low dose strain 19.

Isolation of Brucella abortus from supramammary lymph nodes of cattle from infected herds vaccinated with low dose strain 19.

Synthesis and transudation of antibody during acute inflammation in the mammary gland.

Nonlactating ewes were immunized with a mixture of ovalbumin and tetanus toxoid either in the hind limb, resulting in "priming" of the supramammary lymph node (group L), or in the brisket, resulting in "priming" of the precapsular lymph node (group B). Substantial systemic antibody responses were mounted in both groups. The animals were challenged by intramammary infusion of either tetanus toxoid and lipopolysaccharide (LPS) or ovalbumin and LPS, and changes in the ratios of the two antibody specificities in mammary secretions were monitored for the first 8 h of the resulting acute inflammatory episode. Following challenge with tetanus toxoid and LPS, the ratio of antitetanus toxoid-antiovalbumin in mammary secretion remained close to 1 for the first 6 h postchallenge in both group L and group B. Similarly in animals challenged with ovalbumin and LPS, the ratio of antiovalbumin-antitetanus toxoid in mammary secretion approximated 1 for the first 6 h postchallenge but increased sharply in both groups between 6 and 8 h postchallenge. Measurement of antibody in efferent supramammary lymph suggested that cells in the supramammary lymph node synthesized significant quantities of specific antibody to the infused antigen during the 8-h inflammatory episode.

Comparative effect of selected adjuvants on the response in the bovine mammary gland to staphylococcal and streptococcal antigens

The effect of 4 adjuvants on the response in the lactating bovine mammary gland to an antigenic stimulus was examined. Fifty four lactating Holstein Friesian cows were randomly allocated to 6 groups. Four of these groups received a staphylococcal and streptococcal bacterin-toxoid vaccine administered systemically in association with an adjuvant preparation. The adjuvants used were: aluminum hydroxide gel, Freund's incomplete adjuvant, a metabolizable lipid emulsion and Bordetella pertussis . Two further groups serving as controls received saline, or the vaccine suspended in saline only. The immunoglobulin G response specific for each of 3 vaccine antigens, was monitored in the milk by means of enzyme-linked immunosorbent assay for a period of 23 weeks. The results indicated that high levels of antibody may be maintained in the milk, throughout the average lactation, if cows are vaccinated in the region of the supramammary lymph node with an optimum dose of antigen emulsified in Freund's incomplete adjuvant.

Udder edema of Jersey cattle affected with rectovaginal constriction

Ten Jersey cows affected with rectovaginal constriction (RVC), which developed udder oedema near calving, were studied. One normal Jersey and 3 normal Holsteins served as controls. Before euthanasia the animals underwent daily clinical examination, with special emphasis on the size of the foramen in the rectus abdominis muscle (milk wells). The permeability of the mammary blood vessels was studied by giving one gram of 1% solution Evans blue i/v and examining changes in the colour of the milk and also of interstitial fluid collected by stab incisions. After euthanasia, the entire udder, milk veins, milk wells, subcutaneous tissues and muscles covering the abdomen were removed intact, except in two cows where udder vascular systems were studied by latex infusion. Fifteen minutes after Evans blue injection, the colour of the milk and interstitial fluid were whitish blue and yellowish blue respectively. Oedema involved the entire udder but was more pronounced ventrally. Microscopically there was excessive fluid infiltration between the muscle fibres of the subcutaneous tissues, in the interlobular and interalveolar interstitial spaces of the udder parenchyma, and in the supramammary lymph nodes. The latex studies showed that both milk veins were demarcated by ring-shaped depression of constrictions at the entrance of the milk wells. The studies with Evans blue demonstrated an increase in udder vascular filtration of fluid and protein, possibly due to enlargement or stretching of capillary pores. The latex studies suggested that the milk wells may be a restrictive point for venous blood flow from the udder. Udder oedema may be associated with RVC in Jersey cows.

The influence of site of antigen deposition on the local immune response in the mammary gland of the ewe.

Experiments were carried out to compare the local antibody responses in mammary glands of ewes immunized by infusion of antigen (killed Brucella abortus) into the lactiferous sinuses (trans-epithelial presentation of antigen) or injection into the mammary tissue near the supramammary lymph node (interstitial presentation of antigen). Although both methods of antigen presentation resulted in similar antibody levels in blood, infusion of antigen into the lactiferous sinus resulted in significantly higher levels of agglutinating antibody in milk whey than did injection of antigen. When within-animal comparisons were made, infusion of antigen was also significantly superior to injection of antigen in terms of levels of non-agglutinating antibody in milk as determined by Coomb's antiglobulin assays. Evidence from immunoglobulin estimations in milk whey suggested that any elevation in concentrations of immunoglobulins (including IgA) in milk from ewes, which had received injections of antigen into mammary tissue, was associated with chronic inflammatory damage to these glands.

Smooth phage-resistant Brucella abortus from bovine tissue

Conventional typing, oxidative metabolic, and virulence tests were conducted on a phage-resistant Brucella abortus strain isolated from the supramammary lymph node of a cow.

Immune Response in the Mammary Gland

1.Three classes of immunoglobulins have been identified in lacteal secretions. IgG1 appears to be selectively transferred through the glandular epithelium into the udder. Presumably any method of immunization that stimulates circulating antibodies of the IgG1 class will be beneficial, resulting in IgG1 antibodies in the milk.2.IgA class immunoglobulins are also found in the milk, however, in lower concentration than IgG1 immunoglobulins. IgA is produced locally in the gland since an increase is seen in antigen-infused quarters over noninfused quarters. A factor that might be important is that probably the total amount of any immunoglobulin in lacteal secretions will not be increased by systemic immunization whereas local immunization does increase the amount of IgA.3.The method and route of immunization is possibly very important. Intramuscular, subcutaneous, or other routes that result in systemic stimulation of antibodies, to be effective, must for the most part stimulate class IgG1 immunoglobulins. Hammer, Kickhofen, and Henning (7) have recently published that there is a quantitative shift of antibody activity from IgG1 to IgG2 between the first and third courses of immunization of cattle (7). These IgG2 antibodies would be of little value for mammary gland immunity insofar as antibodies in the milk are concerned. However, since the tissues of the mammary gland are involved in a mastitis infection, specific antibodies of all immunoglobulin classes may be beneficial. Immunization by inoculation of a Streptococcus agalactiae, type II preparation into the area of the supramammary lymph node stimulates protective antibodies in the milk. These antibodies have been shown to be of IgG class principally, with less IgA. Finally, by intramammary infusion of antigen into one or more quarters of a nonlactating gland, an increase in specific antibodies of only the IgA class was noted.4.Progress in the determination of the immunogenic substance necessary to stimulate resistance to staphylococcal mastitis has been slow. Serological typing of staphylococci is not available; however, on the basis of phage typing, most mastitis staphylococci may fall into a dozen or so types (14). Serological typing and the immunogenic response of the group B streptococci are much further advanced. The immunology of the group C and group E streptococci is also in a primitive state. It probably is not necessary to separate and purify each immunogen that is required in a preparation; however, it is necessary that inactivated bacteria of each different antigenic type be present in a bacterin in order to stimulate all the various antibodies necessary.One can speculate that a combination of immunization routes that result in the stimulation of several of the classes of immunoglobulins might produce sufficient protective antibodies in the milk and in the mammary gland tissues to increase resistance. Local stimulation would increase specific IgA while systemic stimulation would result in specific IgG1, IgG2 and IgM. Local stimulation would presumably have to occur in the nonlactating gland with a nonirritating or nontissue damaging antigen. The antigen may be a bacterin consisting of a combination of each of the types of staphylococci and streptococci and be inactivated with formalin or some other inactivant. The bacterin should also contain staphylococcal toxoids.

Ultrasonograms of Lactating Mammary Glands1,2

The use of ultrasound for locating and identifying structures in the lactating mammary gland was explored. A method was outlined for using ultrasound. Internal mammary gland measurements were taken at two different locations; i.e., Spots 1 and 2 being one-third and two-thirds the distance from the floor to the top of the udder, respectively. Change in quarter width as well as median width and gap were compared under five stimulative states—i.e., prestimulation; post-stimulation, premilking; post-normal milking; post-oxytocin injection, precomplementary milking; and post-complementary milking. Sonograms were presented which identified the skin, median and lateral suspensory ligaments, milk ducts, and supramammary lymph nodes. Some general observations were reported on the relation among quarter width, intramammary pressure, and quarter milk production, using limited data. It was shown that following a complete milking, the milk ducts are probably collapsed. The feasibility of using ultrasound to visualize internal structures and measure their responses to vary physiological conditions was demonstrated.

The Cellular and Immune Response in Milk and Regional Lymph During Experimental Mastitis

Experiments have been carried out to study antibody and cellular responses in mammary lymph and milk from ewes with experimental mastitis. Virulent staphylococci were infused into one mammary gland from which lymph was collected for varying periods by cannulation of the efferent mammary lymphatic duct. Anti-α-haemolysin titres rose to relatively high levels in the plasma and lymph, and in the secretion from the infected gland in ewes with acute mastitis, but not in ewes with subacute mastitis. The contribution of the supramammary lymph node to the lymph titre, appeared to be slight. In 2 animals, there was a suggestion of local antibody production by the infected gland. The cellular response in the lymph was characterised by the appearance of polymorphs in large numbers during the first 24. hr. after infection of the gland. In all animals, mature plasma cells in lymph reached a maximum at 5 days, while the maximum antibody titre in lymph occurred 2 days later. In sub-acute mastitis, as signs of inflammation diminished, cellular changes in the milk, similar to those seen during involution of the mammary gland, were observed.