Detection of Salmonella dublin mammary gland infection in carrier cows, using an enzyme-linked immunosorbent assay for antibody in milk or serum
Authors: Smith Bradford P. , Oliver Diane G. , Singh Prithipal , Dilling George , Marvin P. Anthony , Ram Bhanu P. , Jang Larry S. , Sharkov Nikolai , Orsborn John S. , Jackett Karen 
Published: February 2025

SUMMARY
An elisa has been developed for measurement of milk and serum IgG concentrations directed against Salmonella dublin. Four groups of cows were studied: group A—7 experimentally challenge-exposed cows (infected, recovered group); group B—6 normal uninfected randomly selected control cows; group C—7 naturally occurring S dublin carrier cows; and group D—6 normal uninfected S dublin negative cows from the same herd as group C.
Group-A cows were inoculated orally, or inoculated orally and then iv, but none became a S dublin carrier. As expected, all 7 group-A cows responded with a marked increase in elisa titer after oral exposure to virulent S dublin, starting with a mean serum titer of 17.7% and reaching a peak mean serum titer of 79.3% approximately 76 days after initial exposure. As determined by necropsy and organ culturing of the remaining cows, none of the group-A cows became carriers.
The mean serum elisa titer for group-B uninfected control cows was 14.1% (sd ± 12.8%). The mean milk elisa titer was -1.0% (sd ± 5.5%). Colostrum and then milk gave false-positive results for up to 2 weeks after onset of lactation. Group-B cows were culture negative for S dublin in feces and milk during lactation, and when tissues were cultured after euthanasia.
Milk and serum samples for elisa, and milk and fecal samples for culturing were taken from all group-A and -B cows twice a week for 6 months. Statistical correlation (P < 0.05) was found between serum and milk elisa titers. A highly significant (P < 0.001) difference in serum elisa titers was demonstrated between control (group B) and infected cows (group A). Milk and feces from group-C carrier cows were cultured for S dublin 5 days a week for 11 to 13 months. Six of the 7 cows calved during this period. Fecal shedding was sporadic in 7 cows. Milk shedding was frequent in certain quarters of 4 of the cows and was sporadic or absent in other quarters of these cows and it was sporadic in 2 cows, and 1 cow had culture-positive milk only twice. The overall milk-shedding rate was 46% (792 positives/1,733 samples), whereas the overall fecal-shedding rate was 4% (65 positives/1,733 samples). Shedding in the 4 weeks after parturition was 28% in milk and 5% in feces.
Six group-C cows had strongly positive elisa titers in serum and milk, whereas 1 cow (the cow that had only 2 positive milk cultures) had relatively low elisa titers. Group-C cows had a mean serum titer of 85.2% (sd ± 19%) and mean milk titer of 70.6% (sd ± 35.5%). These results indicate that IgG elisa may be useful in detection of S dublin milk shedding (mammary gland infection) carrier cows. Milk shedding in the 4 persistent shedders ranged from 101 to 105 organisms/ml, and was associated with evidence of chronic active mastitis.
Group-D cows, culture-negative herd mates of group-C carrier cows, were monitored in a manner identical to that used for group-C cows. All cows remained culture-negative for S dublin in feces and milk and results of organ culturing were negative for S dublin after euthanasia. The elisa titers remained negative, with a mean group-D titer of 8 ± 7.7% on serum, and 0.6 ± 5.5% on milk. A highly significant difference in serum (P < 0.0001) and milk (P < 0.0001) elisa titers was demonstrated between group-C carrier cows and group-D uninfected herd mates.