Determination of Mycoplasma bovis susceptibilities against six antimicrobial agents using the E test method

The objective of this study was to determine the susceptibility of Mycoplasma bovis against six antibiotics using the E test methodology. Fifty-eight isolates of M. bovis originating from 55 affected cattle were evaluated. Specimen originated from: lung tissue, synovial fluid, tracheo-bronchial wash, milk, and external or inner ear discharge. Antimicrobial agents tested were azythromycin, clindamycin, erythromycin, enrofloxacin, spectinomycin and tetracycline. The E test strips were placed on the surface of Hayflick plates on which organism suspension was spread. Plates were incubated at 35 degrees C in a candle jar for 72 h. MICs were then read by determining where the growth inhibition zone intersected with the MIC scale on the strip. M. bovis Donetta isolate was used as a control. All MICs were >256 microg/ml for erythromycin. MIC50 and MIC90 obtained for azythromycin were 3 and >256 microg/ml, respectively. MIC50 and MIC90 obtained for tetracycline were 4 and 8 microg/ml, respectively. MIC50 and MIC90 obtained for spectinomycin were 2 and >1021 microg/ml, respectively. MIC50 and MIC90 obtained for clindamycin were 0.19 and >256 microg/ml, respectively. MIC50 and MIC90 obtained for enrofloxacin were 0.19 and 0.25 microg/ml, respectively. Resistance was not associated with the specimen source except for azythromycin. M. bovis susceptibilities were easily determined by the E test which demonstrated the efficacy of enrofloxacin and the acquired resistance to tetracycline, spectinomycin, azythromycin and clindamycin.     

Age-related alterations to immune parameters in Labrador retriever dogs

In order to assess age-related changes in the immune status of Labrador retriever dogs, leukocyte phenotypes, lymphocyte proliferative capacity, and serum antibody levels were measured in four cohorts of dogs, ranging from 2 to 10 years of age. Absolute numbers of white blood cells, lymphocytes, monocytes, granulocytes, and CD3+, CD4+, CD8+ and CD21+ lymphocytes significantly decreased with increasing age. Relative percentages of lymphocytes and CD4 cells were significantly decreased, and relative percentages of granulocytes and CD8 cells significantly increased, with age. The CD4:CD8 ratio showed a significant age-related decrease. Proliferative responses of T-cells to mitogens in whole-blood cultures either increased (Concanavalin A) or remained the same (phytohemagglutinin) with age when data was normalised to allow for differences in responding cell number. Similarly, normalised data of proliferative response to anti-CD3 stimulation together with phorbol 12-myristate 13-acetate showed an age-related increase. Serum levels of total IgA significantly increased with age whereas total IgG levels remained unchanged. These observations illustrate a significant change to a number of immune parameters with age. However, further work is required to determine whether the differences reported here are sufficient to cause overt or functional immune senescence in Labrador retriever dogs.     

Activin A: from sometime reproductive factor to genuine cytokine

The growth factor, activin A, was initially characterized as a putative reproductive hormone but is now known to have many other divergent roles. One of these is during inflammation. Following intravenous injection of bacterial lipopolysaccharide (LPS) into sheep, activin A is released extremely rapidly into the circulation. The release of activin A appears to be independent of fever, prostaglandins or other key proinflammatory cytokines such as TNF-alpha or IL-1beta. While the precise roles and function of this factor in inflammation are yet to be elucidated, the activin response occurs in other mammalian species besides the sheep and elevated activin has been documented for a number of clinical inflammatory conditions. Activin A therefore seems to be part of the regulatory component of the innate immune response.     

Relative sensitivity of polymerase chain reaction assays used for detection of feline herpesvirus type 1 DNA in clinical samples and commercial vaccines

OBJECTIVE: To determine relative detection rates and detection limits for 6 published polymerase chain reaction (PCR) assays used for detection of feline herpesvirus type 1 (FHV-1) DNA. SAMPLE POPULATION: 5 vaccines licensed for use in preventing FHV-1-associated disease; 15 conjunctival biopsy specimens collected from cats with keratitis, conjunctivitis, or both; and a plaque-purified field isolate of FHV-1 cultured in vitro. PROCEDURE: Vaccines and clinical samples were assessed for FHV-1 DNA by use of all 6 assays. Detection rates were calculated by assuming that any sample in which FHV-1 DNA was detected was a true-positive result. Detection limits were estimated by use of serial dilutions of DNA extracted from cultured FHV-1 and 1 clinical sample. RESULTS: Testing by use of all 6 assays resulted in detection of FHV-1 DNA in all 5 vaccines. Testing by use of all 6 assays yielded concordant results for 9 of 15 conjunctival biopsy specimens (8 with negative results and 1 with a positive result). Calculated detection rates for clinical samples ranged from 29% to 86%. Assay sensitivity was ranked similarly by use of detection rate or detection limit. CONCLUSIONS AND CLINICAL RELEVANCE: Testing by use of all assays was equally likely to detect vaccine virus. Therefore, a positive PCR result in a cat may reflect vaccine virus rather than wild-type virus. Test sensitivity as assessed by detection limits and detection rates varied greatly. Because FHV-1 can be shed in clinically normal animals, high detection rate will not necessarily correlate with high diagnostic sensitivity.