Monoclonal antibodies that recognize specific antigens of Mycoplasma gallisepticum and M. synoviae

The polypeptide profiles of the type strains of Mycoplasma gallisepticum (PG 31) and M. synoviae (WVU 1853) resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were compared. Except for a few discrete peptides that were similar, the species varied considerably in peptide profiles. Congruence was observed between the type strains of each species and homologous cloned serotypes. Protein blots of each species were probed with 2 mouse monoclonal antibodies. Monoclonal antibody G 46 was specific for the antigen p 110 (G) in M. gallisepticum, and S 221 was specific for an antigen complex p 45-50 (S) in M. synoviae. The 2 monoclonal antibodies clearly distinguished between all serotypes of M. gallisepticum and M. synoviae that were examined by Western blot transfer. Autoradiographs of 125I-labeled M. gallisepticum and M. synoviae indicated that p 110 (G) and p 45-50 (S) were surface membrane peptides. Indirect immunofluorescence of M. gallisepticum and M. synoviae in Vero cell cultures supported the autoradiographic findings. The p 110 (G) antigen of M. gallisepticum was heat-stable, pronase-sensitive, and resistant to periodate oxidation, suggesting that its chemical composition is protein. In contrast, the p 45-50 antigen complex of M. synoviae appeared as a broad band in protein blots treated with monoclonal antibody S 221, was sensitive to pronase, and responded to Schiff's reagent but was not completely inhibited by periodate oxidation, suggesting that it is a complex of repeating sequences probably composed of glycosylated peptides.     

Characterization and activation requirements of bovine lymphocytes acquiring cytotoxic activity after interleukin-2 treatment.

Interleukin-2 (IL-2) treatment of cells and generation of non-major histocompatibility complex (MHC)-restricted cytotoxic cells from peripheral blood mononuclear leukocytes (PBML) was studied. Effector-target conjugate assays demonstrated that bovine PBML bound but did not lyse K562, HL60S and HL60R cells unless activated with IL-2. The magnitude of IL-2-activated killing of tumor cells as well as the magnitude of antibody-dependent cellular cytotoxicity depended on the IL-2 concentration. A short treatment (12-18 h) of effector cells with IL-2 was sufficient for development of cytotoxic activity. Withdrawal of IL-2 from the culture resulted in a reduction of cytotoxic activity that could be restored by further addition of IL-2. Cytotoxic activity of IL-2-activated populations obtained after nylon wool or Sephadex G-10 passage, and Percoll gradient centrifugation of PBML suggests that lymphokine-activated killer (LAK) cell activity in PBML is mainly mediated by a non-adherent lymphocyte lacking markers for B-cells. Positive and negative selection experiments using cell sorting confirmed these findings and demonstrated that the cell responsible for LAK cell activity in cattle belongs to a non-monocyte, non-B, CD2+ lymphocyte population. Furthermore, cytotoxic activity could not be generated in CD2+ populations enriched for cells expressing molecules equivalent to human and murine CD4 and CD8. These findings suggest that effector cells mediating non MHC-restricted cytotoxicity in cattle prevail in a population bearing a CD2+, CD4-, CD8- phenotype and that this population depends on the continuous presence of IL-2 for optimal cytotoxic function.     

Effects of programmed growth rate and days fed on performance and carcass characteristics of feedlot steers.

Two experiments were conducted to investigate a feeding regimen in which a programmed amount of feed was offered daily to control growth rate of steers. In Exp. 1, steers (n = 107, 309 +/- 3 kg) were used to determine effects of offering ad libitum access to feed (AL) vs a programmed intake feeding regimen (PI) and the number of days steers were fed (168 vs 203) on performance and carcass characteristics. Steers in the programmed intake feeding regimen were fed to achieve a predicted gain of 1.13 kg/d for the first 78 kg of gain, 1.36 kg/d for the next 124 kg of gain, and were given ad libitum access to feed for the final 54 or 103 kg of gain before slaughter (for steers fed for 168 d or 203 d, respectively). Feed efficiency was greater (P < 0.02) for steers in the PI than for those in the AL feeding regimen (0.193 vs 0.183 kg gain/kg feed, respectively). From d 169 to 203, steers in the PI feeding regimen had greater (P < 0.06) ADG (1.60 vs 1.38 kg/d) and similar (P = 0.38) feed efficiency than steers in the AL regimen. In Exp. 2, steers (n = 96; 308 +/- 3 kg BW) were offered feed ad libitum throughout the experiment (AL) or were programmed to gain at a high (PI-H) or low (PI-L) growth rate. For the first 78 kg of gain, intake was restricted to achieve predicted gains of 1.13 kg/d (PI-L) or 1.25 kg/d (PI-H). For the next 124 kg of gain, intake was restricted to achieve predicted gains of 1.36 kg/d (PI-L) or 1.47 kg/d (PI-H). Feed was offered ad libitum for the final 58 kg of gain. Overall ADG was similar (P > 0.37) among feeding regimens despite lower DMI for the steers in the PI-L and PI-H feeding regimens than for those in the AL regimen. Feeding regimen did not affect (P < 0.22) carcass characteristics. Programmed intake feeding regimens sustained growth rate and feed efficiency for an extended period of time without detrimental effects on carcass characteristics.     

A paravaccinia virus isolated from cattle.

Isolation of viruses from calves with acute respiratory tract disease were attempted on bovine embryonic lung cell cultures. An isolate obtained from one calf with oral lesions and respiratory disease, designated 44-M-E482, was characterized as a paravaccinia virus on the basis of biological and physical properties. The calf from which the paravaccinia virus 44-M-E482 was isolated did not possess serum neutralizing antibody in its convalescent sera; neither did experimentally inoculated calves possess serum neutralizing antibody to the isolate. However, a low titer of serum neutralizing antibody was produced in one calf after several intravenous injections of the virus. Inoculation of calves with 44-M-E482 into the oral mucosa, skin, nasal cavity and pharynx did not cause any noticeable illness or lesions. The relation of 44-M-E482 to the viruses which cause bovine papular stomatitis and pseudocowpox is discussed.     

Adaptation and negative aftereffect to lateral optical displacement in newly hatched chicks

Chicks wearing hoods containing 8.5-degree wedge prisms from the day of hatching showed both significant reduction in the average lateral displacement of pecking (adaptation) and significant pecking overcompensation in the direction opposite to the original displacement (negative aftereffect) when matched 0-degree plates were substituted for the prisms on the 8th day.     

Evaluation of a commercial enzyme-linked immunosorbent assay for detection of Borrelia burgdorferi exposure in dogs

OBJECTIVE: To evaluate the effectiveness of a commercially available ELISA kit for detecting antibodies against Borrelia burgdorferi in dogs. SAMPLE POPULATION: Banked sera from 440 military working dogs were used for serologic analyses. PROCEDURE: Serum samples were analyzed for antibodies against B burgdorferi by use of a commercially available ELISA and subsequently by western blot analysis as a confirmatory test. RESULTS: Results from the ELISA indicated that 89 (20%) samples were positive for exposure to B burgdorferi or canine Lyme disease vaccine, and 351 (80%) were negative. Follow-up testing by western blot analysis indicated that results for 109 (25%) samples were positive and 331 (75%) were negative for exposure. All samples that had positive results on the ELISA also had positive results on western blot analysis (true positives). Of the 351 samples that had negative results on the ELISA, only 331 had negative results on western blot analysis (true negatives). The remaining 20 samples had positive results on western blot analysis. By use of a standard 2 x 2 table, it was determined that the ELISA had a sensitivity of 82%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 94%. CONCLUSIONS AND CLINICAL RELEVANCE: The commercial ELISA kit evaluated in this study appeared to lack adequate sensitivity for detecting all potential cases of borreliosis in dogs. The ELISA was also unable to discriminate natural exposure from exposure attributable to vaccination, which could complicate interpretation of positive results and treatment of dogs with clinical signs.     

Lung-tissue extract as an alternative to serum for surveillance for brucellosis in chamois

Serological surveys are the most-used way to study diseases in free-ranging wild animals. However, the difficulty in obtaining a sufficient number of suitable serum samples is a major problem. To resolve this problem, we investigated the possibility of using lung-tissue extract in place of blood serum for searching for antibodies against Brucella abortus. Antibodies titres against B. abortus was tested from blood serum and lung-tissue extract from 112 chamois and 99 cattle. Although in complement-fixation-test, lung-tissue extract titres usually were one-to three-fold lower than serum titres, there was a good agreement between serum and lung-tissue extract positivity both in chamois and in cattle. The lung-tissue extract appears a suitable resource in monitoring brucellosis in chamois.     

The status of warning services for plant pests in Italy*

A survey of the present status of warning services for plant protection in Italy shows the lack of a national project, so that the different regional governments approach the question in different ways. In spite of this, some common characters are present: (1) most of the regions manage the warning service directly; (2) everywhere, the warning service interacts with research institutes, farmers’ associations, agrometeorological networks and other warning services; (3)‘indirect warning’ is the prevalent model of organization; geographical areas are divided into homogeneous subareas, where information useful for producing advice is collected and elaborated; warnings are then disseminated by different means of communication, and farmers comply with them autonomously; (4) warnings are usually prepared by a team of advisers, who meet periodically, analyse available information and elaborate suggestions for crop protection; (5) available information comes from field monitoring, weather stations, insect and spore traps, forecasting models for pests and diseases; unfortunately, forecasting models are not widespread; (6) the content of warnings is rather uniform, including information on crops, pests and diseases, suggestions for control strategies and, frequently, meteorological conditions and forecasts; (7) different means are used to disseminate warnings to farmers; usually several methods co‐exist: bulletins published in local newspapers, sent by mail or fax, displayed on notice boards or available via the Internet; placards; telephone messages; surveys on local TV or radio.