We generated monoclonal antibodies (MAB) against feline immunodeficiency virus (FIV) and characterized these MAB by single competition enzyme immunoassays (EIA), immunoblot analysis, and radioimmunoprecipitation. Four MAB identified 3 distinct epitopes of the FIV p24/26 gag major core protein. One MAB recognized the p16/17 gag protein none recognized envelope proteins. We developed an FIV p26 antigen capture EIA that proved more sensitive (0.5 ng of p26/ml), less expensive, and less time-consuming than reverse transcriptase assay. The same MAB were used to develop an antibody EIA specific for FIV p26. The MAB and capture assays reported should prove useful in FIV diagnosis and research.

Marked differences in bone marrow cellularity were observed between cattle affected with leukocyte adhesion deficiency (LAD) and control cattle. The number of nucleated cells in bone marrow was 2.9 to 8.8 times higher in cattle affected with LAD, compared with controls. The myeloid-to-erythroid ratio of bone marrow from 3 cattle affected with LAD ranged from 2.4 to 12. Deficient CD18 expression on neutrophils isolated from bone marrow of cattle with LAD was clearly detected by flow cytometric analysis. Neutrophils from bone marrow of cattle affected with LAD appeared round and not flat, after adherence to plastic wells under agarose, whereas neutrophils from bone marrow of clinically normal cattle were firmly spread on the surface of plastic wells. In the chemotaxis under-agarose assay, many pseudopodia were detected on bone marrow neutrophils from clinically normal cattle, but were not detected on bone marrow neutrophils from cattle with LAD. Activities of chemotactic movements and phagocytosis of neutrophils isolated from bone marrow of cattle affected with LAD were documented to be severely impaired.

Colostrum-deprived calves (n = 34) were fed various amounts of colostrum, colostrum substitute, or milk replacer to establish a range in titer of passively acquired viral neutralizing antibody in serum. The calves were then challenge exposed intranasally with a virulent, noncytopathic bovine viral diarrhea virus (BVDV-890). After viral challenge exposure, calves were monitored for fever, leukopenia, thrombocytopenia, and diarrhea. In addition, viral isolation and viral titration were performed on specimens of nasal secretions, buffy coat cells, and serum obtained from the calves. Fever and systemic spread of virus were detected in calves that had viral neutralizing titer of 256 or lower. Calves that had viral neutralizing titer lower than 16 developed severe clinical disease manifested by fever, leukopenia, thrombocytopenia, and diarrhea. Seventy and duration of signs of disease decreased as titers of passively acquired viral neutralizing antibody increased. These results indicate that low to intermediate titers of passively acquired viral neutralizing antibody were not sufficient to fully protect calves from virulent bovine viral diarrhea virus.

Serum samples from 593 white-tailed deer in Pennsylvania that were killed by hunters in 1991 were examined for Toxoplasma gondii antibodies, by use of the modified agglutination test. Sixty percent (357/593) of the deer had T gondii antibodies; 10% had titer of 25, 23% had titer of 50, and 27% had titer greater than or equal to 500. Sex-specific differences in prevalence were not detected.

Four colostrum-deprived calves each were immunized passively with antisera to whole Pasteurella haemolytica, leukotoxin-containing supernatants of P haemolytica, P haemolytica lipopolysaccharide, or newborn calf serum. Calves were challenge exposed intrabronchially with 5 X 10(9) P haemolytica, and 24 hours later, the resulting lesions were evaluated. The greatest protection against challenge exposure was provided by the antiserum to whole P haemolytica (lesion score = 6.3), whereas newborn calf serum provided the least protection (lesion score = 28.3). Calves that received antiserum to P haemolytica supernatants were moderately protected (lesion score = 16.3), and the antiserum to lipopolysaccharide provided minimal protection (lesion score = 21.8). Antibodies that were unique to whole P haemolytica antiserum and produced dense bands on immunoblots were detected to antigens at 66, 50, and 30 kd. Antibodies in the supernatant preparation that produced prominent bands reacted to antigens between 100 and 90 kd. Collectively, antibodies to these antigens may be responsible for enhancing resistance to experimentally induced pneumonic pasteurellosis. Antibodies to antigens in P haemolytica lipopolysaccharide provided little to no protection.

The effect of oxytetracycline (OTC) on bovine blood mononuclear cells and neutrophil functions was examined in vitro. Neutrophil functions tested include respiratory burst, peroxidase, and antibacterial activities. Neutrophils were treated with OTC (10 to 1,500 > microgram/ml) before exposure to either opsonized zymosan or bacteria. A dose-response inhibition of antibacterial activity to high concentrations of OTC (500 to 1,000 microgram/ml) was observed. Beginning at a concentration of 15 microgram/ml, OTC treatment of neutrophil Iysates resulted in decreased peroxidase activity. A dose response was not observed. In contrast, respiratory burst, measured by nitroblue tetrazolium dye reduction, increased after OTC exposure, but only at high concentrations (500 and 1,000 microgram/ml) of OTC. Mitogen-induced proliferation of blood mononuclear cells cocultured with OTC and concanavalin A, phytohemagglutinin-P, or pokeweed mitogen was inhibited at an OTC concentration of 100 microgram/ml at 48 and 72 hours of culture. These results indicate that blood mononuclear cells are more sensitive to the inhibitory effects of OTC than are neutrophils. Furthermore, the OTC-mediated inhibition of neutrophil antimicrobial activity is inversely related to the increase in nitroblue tetrazolium reduction. This suggests that OTC is uncoupling the hexose monophosphate shunt from production of secreted oxygen radicals. These results also suggest that the peroxidase enzyme system has a large biological reserve capacity.

An ELISA was developed to determine serum concentration of apolipoprotein B-100, a major triglyceride-binding protein in very low-density lipoproteins and a putative maker for hepatic lipidosis of dairy cows. Serum apolipoprotein B-100 was prepared electrophoretically, and antibodies to this protein were raised in rabbits. The antiserum prepared was further purified by affinity chromatography, using bovine serum albumin-Sepharose 4B, to remove antibodies to albumin. For the ELISA, addition of 2-mercaptoethanol to the coating buffer (50 mM sodium carbonate, pH 9.6) was required to evaluate apolipoprotein B-100 concentration in serum. The ELISA developed was sensitive (detection limit was 300 to 400 ng/ml of serum) and reliable (coefficients of variance were in the range of 3.3 to 7.6%). By use of the established ELISA, the serum apolipoprotein B-100 concentration was found to be significantly (P < 0.01) lower during the early lactating stage than during other stages of lactation. Reduced hepatic synthesis or secretion of apolipoprotein B-100 during the early lactating stage, together with the excess uptake by the liver of serum nonesterified fatty acids, is suggested to be relevant in the accelerated accumulation of triglycerides in the liver of dairy cows during the periparturient period.

A rifampicin-resistant Pasteurella haemolytica serotype 1 with 2 added plasmids was used as a colonization-challenge strain in calves to test the resistance to colonization elicited by vaccination. Nine calves were vaccinated with a tissue culture-derived P haemolytica serotype-1 vaccine which, in a prior study, had elicited a serotype-specific inhibition of nasal and tonsillar colonization by the homologous serotype under field conditions. The vaccinates and 9 nonvaccinated control calves were exposed by tonsillar instillation with the challenge strain. The P haemolytica were enumerated in nasal secretion and tonsil wash specimens collected biweekly for 3 weeks. Rifampicin-supplemented agar medium inhibited growth of other bacterial species in the specimens and, thus, increased the sensitivity of detection of the challenge P haemolytica by 100-fold. The challenge strain retained its plasmids during the period of colonization. Inhibition of colonization was evidenced by lower frequency of isolations and fewer isolations of the challenge strain from nasal secretion and tonsil wash specimens of the vaccinates than from those of the nonvaccinates.

The sensitivity and specificity of various serologic tests for antibodies to Toxoplasma gondii were compared in 1,000 naturally exposed sows, using isolation of viable T gondii as the definitive test. Serum samples obtained from heart blood of 1,000 sows from Iowa were examined for T gondii antibodies by use of the modified agglutination test (MAT), latex agglutination test (LAT), indirect hemagglutination test (IHAT), and ELISA. Toxoplasma gondii was isolated from 170 hearts of 1,000 sows by bioassays in mice and cats. The percentage of samples diagnosed as positive for each of the serologic tests was: MAT = 22.2% (titer >greater than or equal to 1:20), IHAT = 6.4% (titer greater than or equal to 1:64), LAT = 10.4% (titer greater than or equal to 1: 64), and ELISA = 24.1% (OD > 0.360). The sensitivity and specificity of these tests were calculated respectively to be: 82.9 and 90.29% for MAT, 29.4 and 98.3% for IHAT, 45.9 and 96.9% for LAT, and 72.9 and 85.9% for ELISA. The dye test was run at 1:20 dilution on only 893 sera because of bacterial contamination and presence of anticomplement substances. Dye test antibodies were found in 17.8% of the sera, and sensitivity and specificity were 54.4 and 90.8%, respectively. Thus, the MAT had the highest sensitivity among all serologic tests used.

Cytologic examination of bronchoalveolar lavage fluid (BALF), including phenotypic analysis of lymphocytes, was performed on 32 Standardbreds with poor race performance and endoscopic examination findings characteristic of inflammatory airway disease (IAD). Nucleated cell counts in BALF from IAD-affected horses were higher than those in control horses; the cytologic profile of BALF in affected horses included mixed inflammation, characterized by mild neutrophilia, lymphocytosis, and monocytosis. Eosinophil and mast cell counts were not higher in the IAD-affected group, compared with those in the control group; however, 4 IAD-affected horses had marked eosinophilia (24.7 +/- 4.8% SEM) in BALF. Phenotypic analysis of lymphocytes in BALF obtained from IAD-affected horses revealed a low proportion of CD4-positive cells and B cells, compared with those in the control group; these findings may have been representative of a greater proportion of non-B, non-T cells (null cells) in horses with IAD. The cytologic profile of BALF obtained from horses with IAD differed from that in horses affected with chronic obstructive pulmonary disease, suggesting that the pathogenesis of inflammation in horses with IAD may differ from that of chronic obstructive pulmonary disease.