The propagation of influenza A (PR 8 strain) and Newcastle disease virus in the bovine mammary gland led to the presence in milk and blood serum of neutralizing antibody. The removal of the gland had a profound effect upon the antibody content of the blood. It is suggested that the major portion of antibody was produced locally in the gland. Whether this results from continuous antigenic stimulation or is a function developed from a fleeting contact with the virus remains unanswered.

Specific immune globulins have been prepared in goat milk in response to the intramammary gland instillation of Neisseria meningitidis 608, serogroup B strain. Isolation, purification, and characterization of the goat whey by gel filtration, electrophoresis, and analytical ultracentrifugation demonstrated that the active immune component resided in the IgA class of globulins, specifically 9.2-S IgA. The potential of the lactating mammary gland as a "biological factory" for the large-scale production of diagnostic antiserum to killed bacterial whole cell antigen is described.

The removal of the udders from two cows known to be infected with Br. melitensis (abortus) was followed in both cases by a reduction in agglutination titre. In the one case, the agglutination titre declined from 1:600 to a value within the negative range, 1:25, and in the other the very high agglutination titre (1:28000) rapidly declined to a point just within the positive range, 1:100.

Goat 111ilk was obtained from the right lactiferous sinus following instillation of PR8 influenza type A virus. The proteins precipitated from goat milk serum by (NH.1)aSO.a were found to contain antibody activity ancl further purification was undertaken. I n the clevelopment of the method the protcin solutions were subjected to paper chromatography and paper and polyucetate high voltage electrophoresis, both before and after a number of isolation a~~d purilication steps. In addition, preparations from each step were run on thc 'Tcclinicon amino acid analyzer for the presence of amino acicls and (or) peptides.As a working procedure, a preparation obtained after cold acetone precipitation was subjected to rivanol treatment, and after electrophoresis a single protein band containing the antibody was obtained. This ~natcrial was further purified by gel filtration using a Bio-Rad P150 column. 'l'he antibod), containing proteln was bnally isolated in the eluate by a fractio~i collector using phosphate buffer as the eluting fluid. There was escellent recovery of the antibody activity In each preparation.

This report describes the development of a method for the isolation of antibodies produced in the mammary gland and found in the milk after the instillation and propagation of various myxoviruses. Biochemical fractionation and isolation procedures have been modified and improved over our previous initial reports. The antibodies of the various influenza and mumps viruses that propagated in the gland were found to be associated with lactogammaglobulin. This report also demonstrates that the antibodies produced in the gland and thus given off in the milk are probably the same as those found in the blood if the animal were infected by conventional routes. Purification of the lactoglobulin protein fraction containing the antibody eliminated the non-specific inhibitors. These results were obtained from the various myxoviruses and mumps that propagated in the goat mammary gland.The main advantages of using the mammary gland as compared to using laboratory animals and their blood are as follows.1. Larger volumes of antibodies can be produced at one time (lactation period) against the influenza and mumps viruses for diagnostic and possibly for therapeutic uses.2. The animal (goat) does not appear to be affected whatsoever by the virus instillation and propagation techniques.3. The milk technique is therefore more humane towards laboratory animals. Invariably the laboratory animal does not have to be sacrificed.