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Isolation and trypsin-enhanced propagation of turkey enteric (bluecomb) coronaviruses in a continuous human rectal adenocarcinoma cell line
1989
Dea, S. | Garzon, S. | Tijssen, P.
Turkey enteric coronavirus (TCV) from intestinal contents of diarrheal poults was isolated and serially propagated in HRT-18 cells, an established cell line derived from a human rectal adenocarcinoma. In these cells, TCV induced cytopathic changes, including polykaryocytosis, which depended on trypsin in the medium and incubation at 41 C. Viral antigens could be demonstrated in the cytoplasm by immunofluorescence, and extracellular virus was detected by an ELISA and negative electron microscopy. The cell-free virus had characteristics of TCV: shape, surface projections, buoyant density of 1.18 to 1.20 g/ml in sucrose, and hemagglutination of rat RBC. The one-step growth curve was complete by postinoculation hours 14 to 16, and maximal titers reached 9 to 9.5 log10 TCID50/ml during 5 passages, after which the titer remained stable. Electron microscopic examination of infected cell monolayers revealed budding of typical coronavirus particles through intracytoplasmic membranes and accumulation of complete virus in cytoplasmic vesicles. Late in the infection, aggregated progeny vial particles were detected near the outer surface of infected cells. One-day-old turkey poults inoculated orally with tissue culture-adapted TCV isolates developed mild to severe diarrhea.
Mostrar más [+] Menos [-]Antigenic and restriction enzyme analysis of isolates of Campylobacter fetus subsp venerealis recovered from persistently infected cattle
1989
Wesley, I.V. | Bryner, J.H.
Thirty-two isolates of Campylobacter fetus subsp venerealis were obtained from 1 bull and 4 heifers with experimentally induced infection. When whole-cell antigens of isolates were cross titrated with antisera to the infecting strain, isolates from 3 heifers had limited antigenic variation, whereas whole-cell antigens of isolates from 2 cattle (the bull and a heifer) differed serologically from those of the infecting strain. Changes were detected specifically in 6 heat-labile antigens. Of the 6 heat-labile factors evaluated, all were initially present on the infecting parent strain, but not on early isolates obtained from 4 of the 5 cattle. Restriction enzyme analysis revealed minor variation in the DNA fingerprints of isolates obtained from individual cattle, thus implying stability of the Campylobacter genome once persistent infection is established. Isolates with identical restriction enzyme patterns expressed different heat-labile antigens. Correlation could not be found between the DNA electrophoretic pattern and the expression of heat-labile antigens.
Mostrar más [+] Menos [-]Prevention of leucaena toxicosis of cattle in Florida by ruminal inoculation with 3-hydroxy-4-(1H)-pyridone-degrading bacteria
1989
Hammond, A.C. | Allison, M.J. | Williams, M.J. | Prine, G.M. | Bates, D.B.
Ruminal microorganisms in cattle at a Florida agriculture research station did not have the ability to detoxify leucaena by degradation of 3-hydroxy-4(lH)-pyridone (3,4,-DHP), but a DHP isomer (2,3-DHP) was degraded in some cattle. Cattle with microorganisms that degraded 2,3-DHP were mostly Senepol cattle imported from St. Croix, US Virgin Islands, where leucaena is an indigenous species. Hereford cattle at the research station in Florida generally did not degrade 3,4-DHP or 2,3-DHP. An experiment was conducted in which a pure culture of 3,4-DHP-degrading bacteria was inoculated into Hereford cattle (with ruminal fistula) grazing leucaena. The bacteria successfully colonized the rumen of recipient cattle and persisted through the following winter when there was no leucaena in the diet.
Mostrar más [+] Menos [-]Effects of inoculations with Eimeria zuernii on young calves treated with decoquinate or narasin with or without dexamethasone
1989
Fitzgerald, P.R. | Mansfield, M.E.
Sixteen 7-week-old Holstein male calves were inoculated with sporulated oocysts of Eimeria zuernii. Four calves (controls) were euthanatized and necropsied at 14 and 20 days after inoculation (DAI). Two calves were treated with 20 mg of dexamethasone (IM) on 13, 14, and 15 DAI and euthanatized and necropsied 17 DAI and 2 calves were given similar treatments and necropsied 20 DAI. The 8 other calves were euthanatized and necropsied 20 DAI. Two were started on the anticoccidial drug decoquinate in feed 13 DAI; 2 others were given decoquinated on the same schedule plus dexamethasone on 13, 14, and 15 DAI. Two calves were given the antibiotic narasin in feed beginning 13 DAI and 2 calves were given parasin on the same schedule plus dexamethasone on 13, 14, and 15 DAI. All calves, except 2 controls necropsied 14 DAI and 4 calves given decoquinate, discharged moderate-to-large numbers of oocysts in feces and had moderate-to-severe changes in fecal consistency. Histologic examintions revealed large numbers of endogenous stages in tissues of calves treated or not treated with dexamethasone. Few endogenous stages were observed in tissues from calves that were given decoquinate or decoquinate plus dexamethasone. Calves given narasin or narasin plus dexamethasone had moderate-to-large numbers of endogenous stages in the tissues.
Mostrar más [+] Menos [-]Role of leukotriene B4 in the pathogenesis of Klebsiella pneumoniae-induced bovine mastitis
1989
Rose, D.M. | Giri, S.N. | Wood, S.J. | Cullor, J.S.
Mastitis was induced in 4 lactating cows by inoculation of Klebsiella pneumoniae (10(7) organisms/ml) via the teat canal. Sterile isotonic saline solution (1 ml) was instilled into designated control quarters via the teat canal. Changes in milk leukotriene B4 and C4 (LTB4, LTC4) concentrations, milk somatic cell counts, and milk bovine serum albumin concentration were monitored over a 24-hour postinoculation period. Milk LTB4 concentration before inoculation in control quarters and quarters later to be infected was 376 +/- 45 and 326 +/- 56 pg/ml of milk, respectively. A significant (P less than 0.05) increase in milk LTB4 concentration in the infected quarters was first observed at postinoculation hour 6, and milk LTB4 concentration in infected quarters generally remained significantly high through postinoculation hour 14. Thereafter, milk LTB4 concentration in infected quarters was not significantly different from the concentration in control quarters. Measurable amounts of LTC4 were not detected in the milk of either control or infected quarters. Milk bovine serum albumin concentration in the infected quarters generally was high throughout the study, as were milk somatic cell counts. The results of this study suggested that LTB4 contributes to the pathogenesis of bovine mastitis.
Mostrar más [+] Menos [-]Efficacy of ivermectin in oral drench and paste formulation against migrating larvae of experimentally inoculated Parascaris equorum
1989
French, D.D. | Klei, T.R. | Taylor, H.W. | Chapman, M.R.
Twenty-one mixed-breed pony foals, reared and maintained under parasite-free conditions, were used to test the efficacy of ivermectin in oral drench and paste formulations (200 microgram/kg) against 11-day-old migrating larvae of Parascaris equorum. Three replicates of 4 foals and 3 replicates of 3 foals were formed on the basis of age. Foals in replicates of 4 were randomly allocated to be indicators, or to receive vehicle (control) or ivermectin paste or ivermectin liquid. Foals in replicates of 3 were randomly allocated to receive vehicle or ivermectin paste or ivermectin liquid. The recovery of larvae from the lungs, liver, and small intestines of the indicator foals showed that 99.9% of the larvae were in the lungs 11 days after inoculation (day 0 of treatment). The recoveries of larvae from lungs and small intestines of controls at 25 days after inoculation indicated that all larvae had migrated to the small intestine by this time. The mean length of larvae recovered from the lungs (11 days after inoculation) was 0.87 mm; the mean length of those recovered from the small intestine (25 days after inoculation) was 3.65 mm. Using larvae recovered from small intestinal contents for calculations, ivermectin in both formulations was 100% effective against 11-day P equorum (P less than 0.01, compared with control group geometric mean of 1498.4).
Mostrar más [+] Menos [-]Endobronchial inoculation of various doses of Haemophilus (Actinobacillus) pleuropneumoniae in pigs
1989
Leengoed, L.A.M.G. van | Kamp, E.M.
Twelve-week-old specific-pathogen-free pigs were inoculated deep in the bronchi with Haemophilus (Actinobacillus) pleuropneumoniae strain 13261 in doses ranging from 8 x 10(1) to 9 X 10(7) colony-forming units (CFU). Pigs that survived infection were euthanatized and examined 48 hours after inoculation. The relationship between dose and severity of disease was evaluated clinically and the weight of pneumonic lesions was compared. The relationship between infection dose and weight of pneumonic lesions proved to be unimodal and not linear. Inoculation of 10(4) CFU of strain 13261 resulted in severe pneumonic lesions and mortality of 29%. In contrast, death was not observed after inoculation with 10(6) CFU of strain 13261 and pneumonic lesions were less severe (P < 0.05). An infective dose of 10(3) CFU induced pneumonic lesions that tended (not statistically significant) to be less severe than those induced by a dose of 10(4) CFU. The peak fever response in all infected pigs was observed from 6 to 12 hours after inoculation. Leukocytosis developed within 12 hours after inoculation, because of an increase of neutrophilic granulocytes. Thereafter, WBC count decreased owing to lymphopenia. Serum iron concentration decreased 80% after inoculation, and zinc concentration decreased 54%.
Mostrar más [+] Menos [-]Infection of the middle nasal meatus of calves with Pasteurella haemolytica serotype 1
1989
Frank, G.H. | Nelson, S.L. | Briggs, R.E.
Eight healthy nonstressed calves were inoculated with Pasteurella haemolytica serotype 1, by instilling a broth culture into the middle nasal meatus of the left nostril. The inoculated left nostrils shed P haemolytica from the ventral nasal meatus at a steady rate for a mean of 7 days, whereas the uninoculated right nostrils of the same calves shed P haemolytica sporadically and in lower concentrations. The duration, frequency, and concentration of P haemolytica shed from the inoculated nostrils was significantly (P less than 0.05) greater than from the nostrils of other healthy calves that had been exposed by instilling the culture into the ventral nasal meatus of both nostrils in a previous study. The concentration of antibodies (IgG, IgA, and IgM) to P haemolytica increased significantly (P less than 0.05) in serum and nasal secretions after exposure. Four weeks after initial P haemolytica exposure, calves were exposed to infectious bovine rhinotracheitis virus and became clinically ill. Four calves were induced to shed P haemolytica from both nostrils by the virus infection; thus, they were harboring the bacterium and were susceptible to active recolonization. Four calves were not induced to shed P haemolytica. The apparent reason was not that they were resistant to active colonization, but that they were no longer harboring the bacterium, because they became active shedders after they were reinfected with P haemolytica.
Mostrar más [+] Menos [-]In situ production of interferon in tissues of chickens exposed as embryos to turkey herpesvirus and Marek's disease virus
1989
Sharma, J.M.
Chicken eggs at embryonation day (ED) 18 or newly hatched chicks were inoculated with turkey herpesvirus (HVT), Marek's disease virus (MDV), or virus-free diluent and, at intervals after inoculation, tissue homogenates of virus-exposed and virus-free chickens or chicken embryos were examined for interferon (IFN) activity. Homogenates of lung thymus and spleen specimens from chickens given HVT at ED 18 had IFN activity. Activity of IFN in the lungs was studied further. Homogenates of lung specimens from chickens exposed to HVT at hatching also had IFN activity, although the concentration of IFN was lower than that in chickens given HVT at ED 18. The pathogenic isolates of MDV (JM-(MDV)), but not the atenuated (Md11/75C-(MDV)) or nonpathogenic (SB1-(MDV)) isolates, inoculates at ED 18 also induced high lung IFN activity. Exposure to a combination of HVT and SB1-MDV induced IFN activity comparable with that in chickens given HVT alone. The IFN activity in homogenates of lung specimens from virus-exposed chickens was species specific and heat and pH stable, but was destroyed by trypsin treatment. Occassionally, low IFN activity also was detected in homogenates of tissus specimens from virus-free chickens or chicken embryos. This IFN activity could have been produced constitutively or may have been induced by substances (inducers) in the environment.
Mostrar más [+] Menos [-]Bovine serum and nasal secretion immunoglobulins against Pasteurella haemolytica serotype 1 antigens
1989
Nelson, S.L. | Frank, G.H.
Experimental intranasal inoculation of cattle with Pasteurella haemolytica serotype 1 resulted in a group that shed the bacteria in their nasal secretions (colonized) and a group that did not shed (uncolonized). After inoculation, antibody titers in serum and nasal secretions against the total P haemolytica increased significantly, and the proportion of total antibody against specific P haemolytica antigens changed so that the proportion directed against the 94- and 62-kD antigens increased. Prior to inoculation, the proportion of total antibody in the serum against 94- and 62-kD antigens of P haemolytica was higher in calves that remained uncolonized than in those that became colonized with P haemolytica after exposure. Antibody specificity of serum and nasal secretions differed in the relative amounts directed against each P haemolytica antigen. The specificity against P haemolytica antigens differed between IgG and IgA isotypes of serum and nasal secretions, with IgA being directed against fewer antigens than was IgG.
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