scholarly journals Experimental Brucella Abortus Strain 19 Arthritis in Young Cattle

1994 ◽  
Vol 6 (1) ◽  
pp. 56-61 ◽  
Author(s):  
B. Johnson ◽  
D. A. Mosier ◽  
R. J. Morton ◽  
A. W. Confer
Keyword(s):  
Brucella Abortus ◽  
Fluorescent Antibody ◽  
Post Inoculation ◽  
Antibody Staining ◽  
Transmission Electron ◽  
Colony Forming Units ◽  
Chronic Synovitis ◽  
Young Cattle ◽  
Culture Negative ◽  
Fluorescent Antibody Staining

Several reports have shown an association between lameness in cattle and vaccination with Brucella abortus strain 19. Affected joints are culture negative for Brucella, but the synovial fluid is positive for B. abortus antibodies. The joints contain cloudy fluid, with villous proliferation of the synovium. Brucella abortus antigens are often found in the synovium with fluorescent antibody staining. This report describes the experimental reproduction of a chronic synovitis in 6 young Angus steers using intra-articular injections of B. abortus strain 19. The carpal and tibial joints were injected with 5 × 109 colony-forming units/ml of B. abortus strain 19 and regularly biopsied over a 28-day period. Steers started becoming serologically positive for B. abortus on post-inoculation day (PID) 5 and were all positive by PID 7. Joints were cultured and examined by fluorescent antibody staining, immunohistochemical methods, and light and transmission electron microscopy. Lesions typical of the field cases were present by PID 21. Brucella abortus was cultured more often during PID 1–5 (6 of 9 joints) than during PID 7–28 (3 of 15 joints). Brucella abortus was only found on PID 1, and 5 by fluorescent antibody staining and in only 2 joints immunohistochemically on PID 5 and 7. The reproduction of lesions typical of field cases but the inability to locate B. abortus antigens in the synovium raises the question of whether in field cases the synovium is continually or intermittently seeded with bacteria or if factors other than just the bacterium are needed to perpetuate the lesion.

scholarly journals EXPERIMENTAL INFECTION WITH MYCOPLASMA PNEUMONIAE (EATON'S AGENT)

1965 ◽  
Vol 121 (6) ◽  
pp. 1071-1086 ◽  
Author(s):  
Adnan S. Dajani ◽  
Wallace A. Clyde ◽  
Floyd W. Denny
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Fluorescent Antibody ◽  
Indirect Fluorescent Antibody ◽  
Quantitative Culture ◽  
Culture Methods ◽  
Qualitative And Quantitative ◽  
Antibody Staining ◽  
Colony Forming Units ◽  
Mycoplasma Pneumoniae Infection ◽  
Fluorescent Antibody Staining

The pathogenesis of Mycoplasma pneumoniae infection was studied in the Syrian hamster with qualitative and quantitative culture methods and special histopathologic techniques. The animals were readily infected with the mycoplasma, which multiplied throughout the respiratory tract. Sensitivity of this experimental host to infection was indicated by the 50 per cent infective dose, which was 10 colony-forming units of the organism. Inoculation consistently resulted in the production of peribronchial pneumonitis which was induced by the mycoplasma. The organisms were visualized in a superficial location in the mucosa of involved bronchi, by means of indirect fluorescent antibody staining and by a modification of the Brown and Brenn technique. The data indicate applicability of the hamster to the study of problems concerned with M. pneumoniae disease which are impractical or impossible to resolve in the human host.

Penetration of Salmonella typhimurium into Turkey Skin

1993 ◽  
Vol 56 (4) ◽  
pp. 292-296 ◽  
Author(s):  
JEONG-WEON KIM ◽  
STEPHEN J. KNABEL ◽  
STEPHANIE DOORES
Keyword(s):  
Salmonella Typhimurium ◽  
Fluorescent Antibody ◽  
Mechanical Damage ◽  
Bacterial Invasion ◽  
Deep Penetration ◽  
Cell Penetration ◽  
Antibody Staining ◽  
Transmission Electron ◽  
Skin Section ◽  
Fluorescent Antibody Staining

Penetration of Salmonella typhimurium into the turkey skins from three different defeathering systems (conventional, kosher, and steam-spray) was quantified by transverse sectioning of inoculated skins with a cryostat and fluorescent antibody staining of skin section homogenates. The microtopography of each skin observed by transmission electron microscopy was correlated with cell penetration. Although few cells were recovered due to freeze injury or freeze killing, direct plating of skin section homogenates showed the different properties of each skin for cell penetration. Fluorescent antibody staining recovered the most cells and revealed the cell distribution within the skin. In conventional skin, over 90% of cells resided in the upper 50 μm depth of skin, which implies the poor penetration of cells. Kosher skin showed relatively even distribution of cells up to 150 μm in depth, which indicated the penetration of cells even into the dermis. Steam-spray skin allowed the deepest (over 200 μm depth) and the highest number of penetration among the three types of skin. Conventional skin retained a thin layer of epidermis (0.3 ~ μm of stratum germinativum) on the surface, and it appeared to act as an effective physical shield against bacterial invasion. The loss of all epidermis on steam-spray skin could explain the deep penetration following this process. Kosher skin retained the most epidermis; however, the characteristic lengthy picking time in kosher processing caused mechanical damage to the skin and allowed deep penetration of cells.

Profilin is predominantly associated with monomeric actin in Acanthamoeba

1999 ◽  
Vol 112 (21) ◽  
pp. 3779-3790 ◽  
Author(s):  
D.A. Kaiser ◽  
V.K. Vinson ◽  
D.B. Murphy ◽  
T.D. Pollard
Keyword(s):  
Monoclonal Antibodies ◽  
Plasma Membrane ◽  
Fluorescent Antibody ◽  
Gel Filtration ◽  
Live Cells ◽  
A431 Cells ◽  
Antibody Staining ◽  
Large Particles ◽  
Biochemical Fractionation ◽  
Fluorescent Antibody Staining

We used biochemical fractionation, immunoassays and microscopy of live and fixed Acanthamoeba to determine how much profilin is bound to its known ligands: actin, membrane PIP(2), Arp2/3 complex and polyproline sequences. Virtually all profilin is soluble after gentle homogenization of cells. During gel filtration of extracts on Sephadex G75, approximately 60% of profilin chromatographs with monomeric actin, 40% is free and none voids with Arp2/3 complex or other large particles. Selective monoclonal antibodies confirm that most of the profilin is bound to actin: 65% in extract immunoadsorption assays and 74–89% by fluorescent antibody staining. Other than monomeric actin, no major profilin ligands are detected in crude extracts. Profilin-II labeled with rhodamine on cysteine at position 58 retains its affinity for actin, PIP(2) and poly-L-proline. When syringe-loaded into live cells, it distributes throughout the cytoplasm, is excluded from membrane-bounded organelles, and concentrates in lamellapodia and sites of endocytosis but not directly on the plasma membrane. Some profilin fluorescence appears punctate, but since no particulate profilin is detected biochemically, these spots may be soluble profilin between organelles that exclude profilin. The distribution of profilin in fixed human A431 cells is similar to that in amoebas. Our results show that the major pool of polymerizable actin monomers is complexed with profilin and spread throughout the cytoplasm.

scholarly journals Detection ofCryptosporidium molnariOocysts from Fish by Fluorescent-Antibody Staining Assays forCryptosporidiumspp. Affecting Humans

2011 ◽  
Vol 77 (5) ◽  
pp. 1878-1880 ◽  
Author(s):  
Rona Barugahare ◽  
Michelle M. Dennis ◽  
Joy A. Becker ◽  
Jan Šlapeta
Keyword(s):  
Ribosomal Dna ◽  
Fluorescent Antibody ◽  
Small Subunit ◽  
Rapid Diagnosis ◽  
Antibody Staining ◽  
Direct Fluorescent Antibody ◽  
Murray Cod ◽  
Diagnosis Of Infection ◽  
Formalin Fixed ◽  
Fluorescent Antibody Staining

ABSTRACTThree direct fluorescent-antibody staining assay kits for the detection of zoonoticCryptosporidiumspecies were used to detectCryptosporidium molnarifrom Murray cod, and the cryptosporidia were characterized by using small-subunit (SSU) ribosomal DNA (rDNA). To facilitate rapid diagnosis of infection, this study demonstrated that all three kits detected freshC. molnariand two kits detected formalin-fixed oocysts.

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