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SUMMARY

The effect of bovine mammary secretion during the early nonlactating period and of antibiotic preparations on bovine polymorphonuclear neutrophil (pmn) phagocytic function and morphology were evaluated in a series of in vitro multifactorial experiments. Benzathine cloxacillin (cl), benzathine cephapirin (ce), sodium novobiocin (no), and a combination of dihydrostreptomycin with procaine penicillin G (dp) were prepared in the presence and absence of a peanut oil aluminum monostearate vehicle. The pmn were isolated from bovine blood, and the effect of each antibiotic preparation on pmn function and morphology was evaluated in a buffer, fat, skim, and a combination of fat with skim from bovine mammary secretion during the nonlactating period. The fat and skim were diluted with buffer to approximate their concentration in mammary secretion. Phagocytic functions of pmn were monitored by fluorescent microscopy, which made it possible to estimate both ingestion and intracellular killing of bacteria by pmn. Changes in pmn morphology were monitored by transmission electron microscopy.

The ability of PMN to ingest and kill Staphylococcus aureus ATCC 25923 was significantly decreased by fat, skim, cl, ce, no, and dp. Effects of some antibiotics on ingestion and killing of bacteria by pmn were influenced by the addition of vehicle and by interactions with mammary secretion. Neutrophil morphology was altered by fat, skim, cl, ce, no, and dp. The detrimental effects of cl, ce, no, and dp on pmn morphology were influenced (some significantly) by the presence of vehicle and interactions with mammary secretion. There were significant correlations among secretion- and antibiotic-induced changes in pmn ingestion of bacteria, pmn killing of bacteria, and pmn morphology.

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in American Journal of Veterinary Research

manual pipetting, washed 4 times with sterile PBSS, a then resuspended in a balanced salt solution d that was supplemented with 10% fetal bovine serum e and 1% penicillin-streptomycin. f Concentration and vitality of PBMCs in each resuspended

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in American Journal of Veterinary Research

5% carbon dioxide). After incubation, nonadherent cells were removed with 1 wash of warm RPMI 1640. Adherent monocytes were overlaid with RPMI 1640 containing 100 U of penicillin/mL and 100 μg of streptomycin/mL. h Control samples used in the study

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in American Journal of Veterinary Research

% heat-inactivated FBS, e 0.1mM nonessential amino acids, f 2mM L -glutamine, g 55μM 2-mercaptoethanol, h and penicillin-streptomycin i (50 U/mL and 50 μg/mL, respectively). After isolation, cells were cultured for 24 hours at 37°C in an atmosphere

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in American Journal of Veterinary Research

microtitration plate. One well received 50 μL of RPMI with 15% FBS and 1.5% penicillin-streptomycin solution as nonantigen-stimulated control, and another well received 50 μL of concanavalin A d (mitogen control) at 0.05 μg/well. The remaining 4 wells received

Full access
in American Journal of Veterinary Research

-glutamine, penicillin (50 U/mL), streptomycin (50 mg/mL), and 5 × 10 −5 M β-2-mercaptoethanol and used immediately in the experimental assays. Macrophage cell line Canine DH82 cells h were used to approximate macrophage function; these cells are

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in American Journal of Veterinary Research

venipuncture with a 20-gauge needle. These samples were placed in blood collection tubes containing sodium heparin. Next, each sample was diluted 1:2 with complete RPMI culture medium (RPMI 1640 with 200 U of penicillin/mL and 200 mg of streptomycin/mL), a and

Full access
in American Journal of Veterinary Research

, M bovis lacks a typical bacterial cell wall. 2 Traditional bacteriocidal antimicrobials, such as ceftiofur and penicillin G procaine, which inhibit bacterial cell wall synthesis, have little effect against M bovis . Therefore, when faced with

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in American Journal of Veterinary Research

humidified chamber at 37°C and 5% CO 2 in IMDM containing 10% HI-FBS, penicillin (100 U/mL), streptomycin (100 U/mL), and tylosin (80 μg/mL). Monocytes were incubated in a humidified chamber at 37°C and 5% CO 2 for 7 days to allow for macrophage

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in American Journal of Veterinary Research

(L929 cells) 31 or chicken myelomonocytic cell line (HD11 cells) 32,33 were seeded at 2 × 10 6 cells/mL onto 6-well tissue culture plates at 37°C and incubated in RPMI 1640 medium a supplemented with 10% fetal bovine serum, b 100 U of penicillin

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in American Journal of Veterinary Research