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.     

Tuberculosis and brucellosis prevalence survey on dairy cattle in Mbarara milk basin (Uganda)

We determined the prevalence of tuberculosis and brucellosis reactors in the dairy herds in the Mbarara district of Uganda in 2002. This is one of the most important dairy-production areas of the country and includes both pastoral and agro-pastoral zones. A total of 340 (of 11,995) randomly selected herds were tested for tuberculosis, using the intradermal tuberculosis-skin test and 315 (of 10,562) herds tested for brucellosis using the serum Rose Bengal test.

The herd prevalence for tuberculosis reactors was 74.1% (95% confidence intervals 69, 78), the individual-animal prevalence was of 6.0% (5.6, 6.5) and within-herd range was 1–50% (up to 100% if suspicious reactors were included).

The herd prevalence for brucellosis was 55.6% (50, 61.2) individual-animal prevalence 15.8% (14.8, 16.7) and within-herd range 1–90%.

The reactor prevalence increased with the age of the animals for both tuberculosis and brucellosis.

Tuberculosis reactor prevalences were higher in animals from the agro-pastoral zone. However, the individual-animal and herd prevalences of brucellosis seroprevalences were higher in the pastoral zone.     

Results of thyroid function tests and concentrations of plasma proteins in dogs administered etodolac

OBJECTIVE: To determine the effects of etodolac administration on results of thyroid function tests and concentrations of plasma proteins in clinically normal dogs. Animals: 19 healthy random-source mixed-breed dogs. PROCEDURE: Blood samples for measurement of serum thyroxine (T4), 3,5,3'-triiodothyronine (T3), free T4 (fT4), and endogenous canine thyroid stimulating hormone (cTSH) were measured twice before as well as on days 14 and 28 of etodolac administration (mean dosage, 13.7 mg/kg, PO, q 24 h). Plasma total protein, albumin, and globulin concentrations and serum osmolality were measured once before as well as on days 14 and 28 of etodolac administration. RESULTS: Etodolac administration did not significantly affect serum T4, T3, fT4, or cTSH concentrations or serum osmolality. Significant decreases in plasma total protein, albumin, and globulin concentrations were detected on days 14 and 28 of administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results of thyroid function tests are not altered when etodolac is administered for up to 4 weeks. Therefore, interpretation of results of these tests should accurately reflect thyroid function during etodolac treatment. Plasma total protein, albumin, or globulin concentrations that are less than the respective reference range in a dog administered etodolac for > or = 2 weeks may be an effect of treatment rather than an unrelated disease process. A decrease in plasma protein concentrations may reflect subclinical injury of the gastrointestinal tract.     

Evaluation of three enzyme-linked immunosorbent assays (ELISAs) for the detection of serum antibodies in sheep infected with Echinococcus granulosus

The aim of this study was to develop an immunological method for the identification of sheep infected with Echinococcus granulosus which would allow the monitoring of animals imported into countries free from hydatidosis and as an aid to countries where control schemes for the disease are in operation. Three enzyme-linked immunosorbent assays (ELISAs) were developed and validated, using as antigen either a purified 8 kDa hydatid cyst fluid protein (8kDaELISA), a recombinant EG95 oncosphere protein (OncELISA) or a crude protoscolex preparation (ProtELISA). Sera used for the assay validations were obtained from 249 sheep infected either naturally or experimentally with E. granulosus and from 1012 non-infected sheep. The highest diagnostic sensitivity was obtained using the ProtELISA at 62.7 and 51.4%, depending on the cut-off. Assay sensitivities were lower for the 8kDaELISA and the OncELISA. Diagnostic specificities were high, ranging from 95.8 to 99.5%, depending on the ELISA type and cut-off level chosen. A few sera from 39 sheep infected with T. hydatigena and from 19 sheep infected with T. ovis were recorded as positive. Western immunoblot analysis revealed that the dominant antigenic components in the crude protoscolex antigen preparation were macromolecules of about 70-150 kDa, most likely representing polysaccharides. This study demonstrated that the ProtELISA was the most effective immunological method of those assessed for detection of infection with E. granulosus in sheep. Because of its limited diagnostic sensitivity of about 50-60%, it should be useful for the detection of the presence of infected sheep on a flock basis and cannot be used for reliable identification of individual animals infected with E. granulosus.     

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.