Important procedural factors in the under-agarose assay for porcine neutrophil migration were identified, and optimal conditions were established. Three factors were tested: the concentration of zymosan-activated serum inoculated into the outer well; the number of neutrophils inoculated into the center well; and the time of incubation of the agarose plates. All factors had a significant (P < 0.0001, 0.0001, and 0.01, respectively) effect on the chemotactic index of porcine neutrophils. The optimal combination of these 3 factors was undiluted zymosan-activated serum as the chemoattractant, 8 x 10(5) neutrophils inoculated into the center well, and 5 hours of incubation. The assay was validated, using standard conditions, and the data were used to predict the number of pigs and/or repetitive assays needed to identify differences among experimental groups.

Neoplastic cells were isolated from 2 sibling Great Dane/Labrador Retriever mixed-breed dogs in which telangiectatic type osteosarcomas arose concurrently. Cells from various sites in the same osteosarcoma appeared similar in culture, but there were differences between the 2 osteosarcomas in growth characteristics and appearance of cells. Cells from 1 osteosarcoma had a small, but significant (P less than 0.05), cyclic adenosine monophosphate response to parathyroid hormone stimulation, indicating a low order of osteoblastic differentiation. Cells from the other osteosarcoma had no response to parathyroid hormone stimulation. Cells from both osteosarcomas and a concentrated cell-free filtrate from the osteosarcoma with osteoblastic differentiation were injected into nude mice, but osteosarcomas were not induced. Results of ultrastructural examination of osteosarcoma samples for viral particles were negative and supernatant fluids from cultured cells were considered negative for viral reverse transcriptase activity.

A study was conducted to examine sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood, including variation attributable to isolation of neutrophils from blood, variation between duplicate determinations of percentage phagocytosis, and the variation in the ability of neutrophils isolated from blood (over repeated collections from the jugular vein) to phagocytose. For the phagocytosis assay, jugular venous blood from each of 4 cows was divided into 2 equal portions. The neutrophils were isolated by lysis of red blood cells with 0.2% sodium chloride. The neutrophils (2 X 10(7)) were incubated in duplicate with 32P-labeled Staphylococcus aureus ([32P]SA; 2 X 10(8)) inskimmed milk samples (2.5% final concentration) prepared from 4 cows. This process was repeated thrice on neutrophils isolated from 4 cows at 2-week intervals. The proportions of variation in percentage of 32P-labeled S aureus phagocytosed between duplicate neutrophil isolations and between duplicate assay determinations were 0 and 1%. Differences among skimmed milk sources and among runs, using blood neutrophils taken at different times from the same donor cow, accounted for 62 and 36% of the total variation. The results indicated that variation arising from blood neutrophil isolation introduced into a phagocytosis assay within a single-day trial is of no concern. The large variation among skimmed milk sample sources indicated differences among cows in the ability of their milk to support phagocytosis. The variation in neutrophil isolations over time for any cow was considered too large to allow for evaluation of physiologic and environmental effects on phagocytosis of neutrophils isolated from blood.

In an attempt to isolate reticuloendotheliosis virus (REV) from field cases, plasma of commercial broiler chickens-suspect to have virus infection-were examined. Samples were inoculated in chicken embryo fibroblasts and after proper incubation infected cultures were assayed for REV-antigen by ELISA, immuno-peroxidase (IP) plaque assay, and PCR.Specificity of ELISA and IP was evaluated by comparing their results with that obtained by PCR. REV could be isolated and virus antigen was detected in cell cultures by all three techniques. Results showed that PCR and ELISA are more specific than IP in detection of REV-antigen. However, the sensitivity of ELISA was affected by the criterion used for determination of the cut-off point. Further studies are needed for full characterization of the isolated virus by using reference antiserum or strain specific primers for PCR.

Two experiments were carried out to evaluate the efficacy of the administration of active dry yeast and/or lactobacillus preparation (AVI-BAC), either before or after the infection with antibiotic resistant field strain of Escherichia coli O127 (E. coli O127) in controlling the severity of infection in quail chicks. The quail chicks of the different experimental groups were infected orally for two successive days with 3x107 CFU of E. coli O-127 as an individual dose. The used field strain proved to be highly pathogenic for quails. Probiotics were supplemented in the drinking water for the different treatment groups at a dose level of 0.5 gm/L. The results revealed that the inclusion of lactobacilli or active dry yeast before E. coli infection has been highly effective in reducing mortality rate, organ invasion and the number of E. coli positive quail chicks. In addition, it decreased the severity of macroscopic and microscopic lesions in different organs in the probiotic treated groups as compared to the infected controls. Lactobacilli preparations were more efficient in controlling the severity of the infection. On the other hand, the administration of yeast and /or lactobacilli after inducing E. coli infection reduced the mortality rate and the severity of lesion score in different organs but probiotics failed to protect quail chicks against the infection. It has been proved that the two probiotics have synergistic effect in controlling collibacillosis in quails.