Production of an equine anti-bovine leukocyte serum.

A method is described for production of an equine anti-bovine leukocyte serum (EABLS). Leukocytes were harvested from the milk of cow's udders which had been irritated with endotoxin. The washed leukocytes as antigens were administered to 2 horses in a series of subcutaneous and intravenous injections. There was a variably progressive increase in total serum proteins and a decrease in albumin/blobulin ratios, but the most pronounced change was an increase in beta2-globulins. Accompanying these changes was an increase in the number of precipitin lines as shown by Ouchterlony analysis. Four old cows and 2 calves were intravenously given a 50- and 20-ml dose, respectively, of the EABLS. Two of the cows and 1 of the calves were given the EABLS which had been heated to 56 C for 30 minutes. Both calves and the 2 cows given heated EABLS and 1 cow given unheated EABLS developed marked neutropenia within the 1st hour. The 4th cow given unheated EABLS developed only slight neutropenia. It was concluded that there may be variation in the individual animal's susceptibility to the neutropenia-inducing activity of EABLS.

Effect of halothane and halothane-nitrous oxide on hematocrit and plasma protein concentration in dog and monkey.

Hematocrit and plasma protein concentration in healthy dogs and monkeys (Macaca arctoides) awake and anesthetized with halothane-oxygen and halothane-nitrous oxide oxygen were compared during conditions of spontaneous and controlled ventilation. Both hematocrit and plasma protein concentration decreased within 15 minutes following anesthetic induction. This decrease persisted throughout constant- or variable-depth anesthesia and did not vary appreciably with ventilation, anesthetic dose, or introduction of nitrous oxide. Plasma volume, determined by a dye dilution technique, concomitantly increased. This increase is compatible with the directional changes in hematocrit and plasma protein.

Conversion of chronic staphylococcal mastitis to acute gangrenous mastitis after neutropenia in blood and bone marrow produced by an equine anti-bovine leukocyte serum.

A stock strain of Staphylococcus aureus of mastitis origin, characterized by alpha-, beta-, and delta-toxins, was used to produce chronic mastitis of 20 to 300 days' duration in 6 lactating mammary quarters of 4 cows. Early acute Streptococcus agalactiae mastitis was produced in 1 additional mammary quarter of 1 cow. Equine anti-bovine leukocyte serum (EABLS) was administered to all cows by continuous intravascular drip for 12 to 32 hours. Neutropenia in blood and partial depletion of neutrophil reserve in bone marrow were produced. Chronic subclinical staphylococcal mastitis in 2 quarters of 1 cow changed to gangrenous mastitis by the 40th hour after EABLS administration and led to death of the cow. The disappearance of neutrophil leukocytes from the milk was followed by uninhibited multiplication of S aureus. Probably, staphylococcal leukocidins accelerated the destruction of neutrophils in the milk as S aureus multiplication became intensified. In another quarter of the same cow that was infected with Str agalactiae, neutrophil leukocytes were present in milk as long as 3 days after their disappearance from blood and bone marrow. This may give some indication of the extravascular life-span of the neutrophil in the udder in mastitis. The 2nd cow died at the 16th hour from the start of EABLS administration and at a time when gangrenous mastitis was in the initial stages of development. The S aureus-infected quarters of the 2 remaining cows did not become gangrenous. Administration of EABLS to these 2 cows did not significantly reduce the numbers of neutrophil leukocytes entering the milk of the 3 S aureus-infected quarters. It is concluded that continuous diapedesis of neutrophil leukocytes into the milk in chronic staphylococcal mastitis protects the gland against the development of gangrenous mastitis in the presence of a strain of S aureus capable of alpha-toxin production.

Cytologic features of bone marrow in normal and mastitic cows.

Rib bone marrow was aspirated from normal cows and from cows with experimentally induced or naturally acquired mastitis. Nucleated cell counts in marrow films were made on the basis of differentiation of 500 cells. The myeloid/erythroid ratio was less than 1.0 in normal cows and generally greater than 1.0 in mastitic cows. The marrow reserve of mature neutrophils was nearly depleted in response to experimentally induced Streptococcus agalactiae mastitis (4th postinoculation day) or in an acute flare-up of naturally acquired Str agalactiae mastitis. A dose of 0.05 mg of Escherichia coli endotoxin, introduced into a single, normal, lactating quarter produced similar results within 8 hours. Concurrently, with depletion of marrow reserves of mature neutrophils, a neutropenia with left shift developed in the blood, and immature granulocytes increased in the marrow. The intensification of granulopoiesis returned the marrow reserves to normal or above normal within a period of 4 to 5 days, thereby reestablishing homeostasis.