Detection and characterization of leptospiral antigens using a biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay and immunoblot.

A biotin/avidin double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of antigens of Leptospira interrogans serovars in experimentally inoculated bovine urine samples was evaluated. Immunoglobulin G (IgG) from rabbits immunized with L. interrogans serovar hardjo type hardjobovis sonicated, whole cell, and formalinized-heated antigen preparations were purified by a protein A-superose column coupled to fast protein liquid chromatography, and evaluated for species specificity in the ELISA. The ELISA using each specific IgG detected as few as 10(4) leptospires of the homologous serovar hardjo diluted in phosphate-buffered saline solution with Tween 20 (PBSS-Tween 20). On immunoblot analysis of proteinase-K-digested whole cell leptospiral preparations, each IgG revealed the presence of bands specific to serovar hardjo, suggesting the presence of serovar-specific epitopes on the lipopolysaccharide molecules. The minimum number of cells of heterologous serovars pomona, grippotyphosa, bratislava, icterohaemorrhagiae and copenhageni detected by each ELISA was greater, ranging from 10(6) to 10(7). The common antigenic determinants observed on immunoblot analysis were different for each specific IgG, except for a major cross-reacting, possibly flagellar, protein doublet at approximately 36-36.5 kDa. Leptospires were equally well detected by the ELISA in both bovine urine and PBSS-Tween 20.     

Identification by isoelectric focusing of creatine kinase-MM isoforms in the plasma and striated muscle of pigs

Total creatine kinase (CK) and CK-MM isoforms were determined in plasma and longissimus dorsi muscle extracts from normal pigs. Based on their total CK activity, the pigs were divided into two groups. Pigs of group 1 (n=16) had a mean plasma total CK of 298±16 U/L and the distribution of the CK-MM isoforms was 65.7±2.5% CK-MM₃, 18.9±1.6% CK-MM₂ and 15.3±1.5% CK-MM. In group 2 (n=18; 826±75 U/L total CK) four isoforms were observed: 3.1±0.9% CK-MM, 67.9±3.0% CK-MM₃, 21.5±2.3% CK-MM₂ and 7.5±1.3% CK-MM₁. The differences between the two groups of pigs were significant (p<0.001) for CK-MM₁ and the presence of CK-MM. Four CK-MM isoforms were also detected in longissimus dorsi muscle homogenates: 45.6±8.1% CK-MM, 32.6±11.7% CK-MM₃, 16.6±2.3% CK-MM₂ and 5.1±2.8% CK-MM₁. The release of CK-MM isoforms from muscle into plasma seems to be unrelated to the concentration of these isoforms in striated muscle.     

Humoral immune responses of black-footed penguins (Spheniscus demersus) after DNA-mediated immunization with a beta-galactosidase reporter gene.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a beta-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-beta, containing the beta-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-beta-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to beta-galactosidase were detected in the sera of pCMV-beta-inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding beta-galactosidase and will mount a humoral response against the in vivo-expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.     

STUDIES ON THE MANAGEMENT OF CARDIOPULMONARY EMBARRASSMENT DUE TO THIOPENTONE ANAESTHESIA IN THE EQUINE

Cardio pulmonary embarrassment was induced with thiopentone sodium in 35 healthy adult donkeys divided in to seven groups of five animals. “To effect” anaesthetic dose was (12.72 ± 0.69 mg/kg) which caused transient hypotension, tachycardia and hypoxemia. A stable fall in MAP 1. 6.77 ± 0.287 Kpa (50.78 ± 2.15 mmHg) and a flat EEG were considered to indicate acute circulatory insufficiency due to thiopentone overdose.     

Effects of monensin on Mg, Ca, P and Zn metabolism and tissue concentrations in lambs

A study consisting of two trials was conducted to determine the effects of monensin on the apparent absorption and retention of magnesium (Mg), phosphorus (P), calcium (Ca) and zinc (Zn) and to determine mineral changes in tissue and ruminal fluid. Eight lambs (39 kg) were used in trial 1, and 10 lambs (37 kg) were used in trial 2. Animals were blocked by weight and fed a high concentrate diet with or without 20 mg/kg monensin. Trials began with a dietary adjustment period lasting 18 d in trial 1 and 21 d in trial 2. Animals were then placed in metabolism stalls for a 10-d stall adjustment period followed by a 12-d collection period. Collections to determine mineral balance were made during the first 10 d of the collection period. Blood and ruminal fluid samples were taken on d 11 of the collection period. Lambs were slaughtered on d 12 of the collection period and tissue samples were collected. Monensin supplementation increased (P less than .05) Mg retention 42.0%. Urinary Ca excretion decreased (P less than .05) 60.0% when monensin was fed. Monensin supplementation decreased (P less than .05) liver Ca and bone Ca, 45.5 and 2.9%, respectively. Apparent P digestibility increased (P less than .05) 40.0% and P retention increased (P less than .10) 26.8% due to monensin supplementation. Both apparent absorption and retention of Zn increased (P less than .01) 50.0 and 45.0%, respectively, with monensin supplementation. Ruminal fluid Zn concentrations decreased (P less than .05) 33.0% with the addition of monensin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevalence of leukemic blood and bone marrow in dogs with multicentric lymphoma

Multicentric lymphoma was diagnosed in 53 dogs. A study was performed to evaluate the prevalence of leukemic involvement in blood samples, bone marrow aspirates, and bone marrow core biopsy specimens at the time of initial diagnosis. Data indicated that 57% (30/53) of the dogs were leukemic when all materials were considered relative to the presence of cellular atypia or immaturity and abnormal tissue distribution. In the 30 leukemic dogs, detection was made in the specimens with the following frequency: 15 in blood (50%), 18 in bone marrow aspirates (60%), and 29 in bone marrow core biopsy specimens (97%). Five cases (17%) were only detected by core biopsy examination, even when dogs with bone marrow lymphocytosis of greater than 15% of nucleated cells were considered leukemic. Nondiffuse histologic colonization patterns accounted for the lack of correlation between the type of bone marrow specimens. Clinical staging for treatment response and prognosis was best determined by evaluation of concurrently obtained blood samples, bone marrow aspirates, and bone marrow core biopsy specimens.     

Environmental source of mycobacteriosis in a California swine herd.

Between July 1985 and April 1986, mycobacterial lymphadenitis was recorded in six of 2407 slaughter pigs from a commercial swine herd in which the majority of pigs were raised in confinement. Records showed that all six affected pigs had access to dirt-floored pens at least 81 days prior to slaughter. The mycobacteriosis lesion rate for pigs exposed to dirt pens was 9.4% while in nonexposed pigs the lesion rate was zero. The risk associated with movement of pigs from concrete floored pens to dirt pens was evaluated by a field trial. In the field trial, two litters (5 of 15 pigs) exposed to dirt pens at 12-24 days of age but none of nine nonexposed litters (39 pigs) developed lesions. Mycobacterium avium-complex bacteria were recovered from both exposed litters (9 of 15 pigs) but from none of nine nonexposed litters. Serovars of M. avium-complex isolated from trial pigs included 1, 4, 8, 9, the dual serovar 4/8, and an untypable serovar. Incense-cedar bark (Calocedrus decurrens) used as a flooring material in the pens was demonstrated to be a potential source of M. avium-complex serovar 9. The dual serovar 4/8 and an untypable M. avium-complex were isolated from the dirt-floored pens. No evidence of cross-transmission of M. avium-complex infection was detected. The sporadic pattern of mycobacteriosis observed in the herd probably resulted from infrequent exposure to a common environmental source.     

The effect of Harderian adenectomy on the antibody response in chickens

Intact chicks and those that had their glands of Harder (GH) removed (GHx) at 1 day of age were studied for their response to optically or intraperitoneally (IP) applied antigens. Following exposure of the chicks to sheep red blood cells (SRBCs), killed Brucella abortus, or bovine serum albumin (BSA), serum and tear samples were collected and assayed for antibodies. Of the two sources of antibodies, the serum generally had higher levels than did the tears. The only exception to this occurred in the intact chicks inoculated by the eye, in which serum and tear levels were equivalent. With SRBCs, no difference could be detected between the two routes of inoculation. However, IP inoculation produced higher levels of antibody in the serum of intact and GHx chicks inoculated with B. abortus or BSA and in the tears of the GHx chicks exposed to B. abortus. Removal of the GH resulted in a consistent decrease in antibody levels in the tears, regardless of the route of exposure. Although this effect was noted with all three antigens, it was more pronounced in the trials using B. abortus and BSA. This finding is discussed in terms of describing the importance of the GH as a source of antibodies to optically applied antigens, and its importance as a route of circulating antibody egress. Furthermore, the feasibility of using the antibody response in tears to a test antigen is discussed as a means of measuring the immune status of a functioning GH.