We investigated the ability of an antibody-specific, O antigen-based ELISA to document Salmonella typhimurium herd infections by screening of milk samples. Three cattle populations, 20 herds with no history of salmonellosis, 8 herds with history of S. typhimurium episodes within the previous 7 months, and 220 herds of unknown disease status, were tested. A herd was considered ELISA positive if at least 5% of the cows had OD values > 0.3. Among the 20 herds without history of salmonellosis, only 2 herds were ELISA positive, whereas all 8 herds with a known history of salmonellosis were ELISA positive (herd specificity, 0.9 and herd sensitivity, 1.0). A significant correlation (P < 0.001) was found between the OD values of serum and milk samples from cows in the herds with a history of salmonellosis. It was concluded that ELISA testing of individual milk samples can be used for surveillance of herds for S. typhimurium infections, but further modifications are needed to test bulk tank milk samples.

Introduction: Abortion in cattle is a major source of economic losses for the agriculture sector. It can be due to infectious or non-infectious factors. Among infectious factors, parasites, bacteria, viruses, and fungi can be involved. The present work investigated the prevalence of the main infectious agents of abortion in Algerian cattle. Material and Methods: Altogether 278 non-aborting and 82 aborting cows were analysed. Results: The prevalence ranged from 0% for Tritrichomonas foetus to 15% for Neospora caninum. Additionally, a case-control study was performed to find the association between the presence of the pathogens and the occurrence of abortion in cows. The odds ratios were significant for Neospora caninum, bovine herpes virus 4, BVD virus, Brucella abortus, Salmonella Dublin, Leptospira interrogans serovar Hardjo, and Coxiella burnetii. Conclusions: The pathogens enumerated here could be major causes of abortion among Algerian cattle.

We investigated the ability of an antibody-specific, O antigen-based ELISA to document Salmonella typhimurium herd infections by screening of milk samples. Three cattle populations, 20 herds with no history of salmonellosis, 8 herds with history of S. typhimurium episodes within the previous 7 months, and 220 herds of unknown disease status, were tested. A herd was considered ELISA positive if at least 5% of the cows had OD values > 0.3. Among the 20 herds without history of salmonellosis, only 2 herds were ELISA positive, whereas all 8 herds with a known history of salmonellosis were ELISA positive (herd specificity, 0.9 and herd sensitivity, 1.0). A significant correlation (P < 0.001) was found between the OD values of serum and milk samples from cows in the herds with a history of salmonellosis. It was concluded that ELISA testing of individual milk samples can be used for surveillance of herds for S. typhimurium infections, but further modifications are needed to test bulk tank milk samples.

Cows and calves from a 1,600-cow drylot dairy were screened for IgG antibodies to Salmonella dublin lipopolysaccharide (LPS), using an indirect ELISA. The ELISA was performed on milk samples from lactating cows and on sera from nonlactating cows and calves. Fecal samples were collected from calves and nonlactating cows for culture of Salmonella spp. All seropositive cattle were retested by culture and ELISA 5 times at monthly intervals or until antibody concentration decreased. None of the cattle remained culture-positive and seronegative. Prior to and during the sample collection period, approximately 30% of calves < 8 weeks old died of S dublin infection. Vaccination of cows with a killed S dublin/S typhimurium vaccine at cessation of lactation was a routine management practice. The ELISA-determined Igg response to vaccination had decreased by 50 days after vaccination. Eight cows and 5 calves that maintained a high serologic response to S dublin were purchased and moved to a research facility for 6 months of intensive monitoring. Lactating cows were milked twice daily, and culture of milk and feces for Salmonella spp was performed 5 times/wk. Serum IgG antibodies to S dublin LPS were measured weekly, using ELISA. At the end of 6 months, all 13 cattle were necropsied and tissues were obtained for culture of Salmonella spp. All 8 cows and 5 calves maintained persistently high ELISA titer for the 6 months of testing, and shed S dublin in the milk and/or feces during the same period. On this basis, they were termed S dublin carriers. Salmonella dublin was isolated from mammary tissue of 2 calves at necropsy, indicating that bacteremia may be a mode of mammary infection by S dublin. Results of the study indicated serologic testing can be used successfully on a large dairy to identify S dublin carrier cattle. Using initial milk screening, 42 of 1,268 lactating cows were identified as suspect, requiring repeated serologic testing. One nonlactating cow, 7 of the 42 suspect lactating cows, and 5 of the 222 calves maintained an Igg response, and were found to be S dublin carriers. Carrier cows shed S dublin in 3.35% of fecal samples and 2.51% of milk samples, and carrier calves shed S dublin in 17.26% of fecal samples.

Immunoglobulin reactions to Salmonella dublin in serum and milk from 4 groups of lactating cows were measured by an indirect ELISA. The groups consisted of (1) cows that were natural carriers of S dublin in the mammary gland, (2) experimentally infected cows that did not become carriers, (3) cows inoculated with a commercial S dublin bacterin, and (4) cows used as S dublin-negative controls. Milk and serum samples were obtained at monthly intervals. Models for predicting carrier status were developed by use of stepwise logistic regression. Independent variables consisted of serum and milk IgG and IgM titers to S dublin lipopolysaccharide and a ratio of IgG to IgM. The utility of a single sample vs multiple samples obtained at 1-month or 2-month intervals was tested by comparison of goodness-of-fit X2 P values for 8 models predicting carrier status. Immunoglobulin reactions specific to S dublin were a significant predictor of carrier status (P < 0.001). Serum IgG titers specific for S dublin were the most important variable for predicting carrier status. Two serum IgG titers to S dublin obtained 2 months apart was a better predictor of carrier status than measurement of the IgG:IgM ratio from a single serum sample. Immunoglobulin recognizing S dublin epitopes also were detected in milk samples. In milk, performing 2 ELISA 60 days apart to determine IgG and IgM reactions to S dublin appeared to be useful for the prediction of carrier status, but was not as accurate as models for serum immunoglobulin reactions.

Serologic testing to detect persistent IgG titer directed at Salmonella lipopolysaccharide (LPS) has proven useful in detecting Salmonella carrier cattle without clinical signs of disease and in seroepidemiologic studies. Although little cross-reactivity exists between most Salmonella serogroups, groups B (O1, 4 [5], 12) and D (O1, 9, 12) share somatic (LPS cell wall) antigens O1 and O12, which results in some cross-reactions. This may be unimportant in most instances, because group-B and group-D carriers need to be identified and culled. It may be desirable in some situations, such as when trying to control S dublin, to determine which serogroup is present in a given herd. For this reason, a procedure to produce a pure O9 group-D antigen was developed. Salmonella dublin (group D) was grown by use of standard procedures, and LPS was extracted by use of the phenol-water method. The LPS was then oxidized with sodium periodate, dialyzed, reduced with sodium borohydride, cleaved with hydrochloric acid, and again dialyzed. This procedure successfully cleaved the saccharides comprising O antigens 1 and 12, leaving a pure O9 ELISA antigen. Sera from cattle vaccinated or naturally infected with S typhimurium, S agona, and S schwarzengrund (all group B), S montevideo (group C1), and S dublin (group D) were tested by ELISA, using modified and unmodified antigens. When the ELISA antigen used was the chemically modified (pure O9) group-D antigen, elimination of cross-reactions confirmed the structural loss of cross-reacting O1 and O12 antigens.

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 less than 0.05) was found between serum and milk ELISA titers. A highly significant (P less than 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 10(1) to 10(5) 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 less than 0.0001) and milk (P less than 0.0001) ELISA titers was demonstrated between group-C carrier cows and group-D uninfected herd mates.

Studying of efficiency of application of modified scheme of hyperimmunization of bull-producers with a designer salmonella polyantigenin in comparative analysis with the production scheme of hyperimmunization was realized in the conditions of the Republic of Belarus. Research results showed that engineered polyvalent antigen for hyperimmunization of producer bulls contained in their formula the following serovariants of salmonella in 1:1 ratio: S. typhimurium 371, S. dublin 373, S. choleraesuis 370, and S. enteritidis CB. The obtained antigen was sterile, harmless and had pH 7,3. Hyperimmunization of producer bulls in the experimental scheme revealed in increasing of leukocytes quantity on 57,3 %, banded neutrophils on 86,7 %, B-lymphocytes - on 42,2 %, content of total protein - on 25,6 %, content of Ig G - on 77,8 %, lg M - on 51,6%. After the fifth administration of antigen there was stated the lowering of agglutinating activity of blood serum on 25%, content of Ig G - on 54,4%, Ig M - on 14,6 %, that indicated to the inefficiency of antigen administration and became the basis for termination of hyperimmunization cycle. Immunobiological reaction of bull-producers which were hyperimmunized by the suggested scheme differed from reaction of animals which were hyperimmunized by the production scheme: content of total protein increased on 10,1%, protein content - on 27,3 %, content of lg G - on 24,8 %, lg M - in 23,7%

Experimental research on production of antigen preparation for hyperimmunization of bulls from blood of which it was possible to produce an active medical and preventive serum against cattle salmonellosis and pasteurellosis was realized in the conditions of the Republic of Belarus. There were developed four variants of antigen on the basis of the formolated aluminous concentrated vaccine against calf salmonellosis and on the basis of semiliquid aluminum hydroxide vaccine against cattle pesteurellosis. All obtained variants of associated antigen had concentration of hydrogen ions, which was close to the neutral rating; they were also sterile and safe for white mice. Associated antigen which was constructed from vaccines in the ratio of 1:1, 1:2, 1:3, and 2:3 had an ability to raise the protection of immunized experimental guinea pigs in the conditions of controlled introduction of infection with S. dublin 373 and S. tythimurium 371. Pasteurellosis component of associated antigen in 1:1 and 1:2 variants protected against death only 3 out of 5 immunized doves, both in relation to P. multocida strain N 796, and bacteria P. multocida strain N 5264. Vaccine ratio in the associated antigen 1:3 and 2:3 turned to be more immunogenically balanced: all tested doves survived after introduction of infection of broth culture P. multocida of N 796 and N 5264 strains. Experimental results showed that the associated antigen on the basis of vaccines in ratio of 1:3 was sterile, safe, active and suitable for hyperimmunization of bull producers serum against salmonellosis and pasteurellosis of calves, as well as antigen in the variant 2:3, but this ratio increased the expenses