Detection of urogenital mycoplasmal infections in primates by use of polymerase chain reaction.

Urogenital mycoplasmal infections could affect use of primates as models for reproductive system studies and could affect reproduction in captive primates, but could be useful as animal models of similar human infections. We conducted a pilot study to assess detection of urogenital mycoplasmal infections in primates by use of polymerase chain reaction (PCR). Healthy animals were anesthetized, and vaginal, cervical, or endometrial and urethral swab specimens were collected from females and males, respectively. Specimens were tested by PCR supplemented with dot blotting and nonradiolabeled oligonucleotide probing for 16S rRNA sequences conserved among mollicutes. Specimens with positive results were tested by species-specific PCRs with primers for 16S rRNA sequences of Ureaplasma urealyticum and Mycoplasma hominis and for adhesin gene sequences of Mycoplasma genitalium. Spiked duplicate reactions were included as internal controls for each reaction. Results for 232 specimens from 166 animals indicate that naturally acquired urogenital infections are readily detected and suggest that urogenital mycoplasmal infections are common in laboratory primates (48/166 [29%] overall). M. hominis and U. urealyticum appeared to be common among the studied primates overall and especially in chimpanzees. Mycoplasmas other than M. genitalium, M. hominis, and U. urealyticum appeared to be at least as common as these three, with specimens from 18 of 48 animals (38%) having positive "generic" PCR results, but no positive results in species-specific PCRs.

Detection of Mycoplasma pulmonis in cilia-associated respiratory bacillus isolates and in respiratory tracts of rats by nested PCR.

To improve the detection of Mycoplasma pulmonis contamination of isolates of cilia-associated respiratory (CAR) bacillus, we developed a nested PCR method using primers for 16S rRNA gene sequences. Of 140 samples of 16 different CAR bacillus isolates, 73 (52%) were inhibitory in the first PCR, as indicated by the absence of amplicons of the internal control, but only 11 of 140 (7.9%) were inhibitory in the second PCR. Of 27 samples known to contain M. pulmonis, only 12 (44%) were positive in the first PCR, but 25 of 27 (93%) were positive in the second PCR. Nested PCR also detected M. pulmonis in 21 of 61 (34%) CAR bacillus samples from which M. pulmonis could not be cultured and identified 2 additional M. pulmonis-contaminated CAR bacillus isolates. Of 359 respiratory and reproductive tract lavage samples from rats and mice, 35 (9.8%) were inhibitory in the first PCR, but only 15 (4.2%) were inhibitory in the second PCR. Of 72 lavage specimens from rats inoculated with an avirulent, poorly infective M. pulmonis strain, 14 (19%) were positive by nested PCR, but only 2 of 72 (2.8%) were positive by culture. Nested PCR also detected M. pulmonis in 14 of 20 (70%) paraffin sections of lung and trachea from rats and mice inoculated with CAR bacillus isolates known to contain M. pulmonis, whereas single PCR gave no positive results. We conclude that nested PCR is superior to single PCR or culture for detecting M. pulmonis, and that M. pulmonis is present in all but four CAR bacillus isolates in our collection that were from naturally infected rats; the four isolates that were exceptions were obtained from rats from a single colony.

Adverse side effects of oral antibacterial therapy in dogs and cats: an epidemiologic study of pet owners' observations.

This epidemiologic study surveyed pet owners for observations of adverse reactions to oral antibacterials which were prescribed to outpatients at a university teaching hospital. Multiple questions were asked of all clients whose dog or cat was treated during the 18-month study. The questionnaire was designed prospectively to define those reactions that clearly seemed to be medication related. In this study, owners noted a significant number of adverse side effects with erythromycin stearate administration to dogs. Trimethoprim-sulfadiazine was used most often in dogs but did not result in a substantial number of adverse reactions.

Recognition of triiodothyronine-containing epitopes in canine thyroglobulin by circulating thyroglobulin autoantibodies.

Serum triiodothyronine autoantibody (T3 AA), triiodothyronine (T3), and thyroxine (T4) concentrations were determined in 45 canine sera containing substantial amounts of thyroglobulin autoantibodies (Tg AA); sera also were assayed to investigate the ability of free T3 to inhibit Tg AA binding to canine Tg. Serum T3 AA concentrations defined 2 groups of sera; 28 sera had low T3 AA concentration (< or = 20 ng/ml) and 17 sera had high T3 AA concentration (> or = 250 ng/ml). Direct linear correlation between T3 AA concentration and apparent serum T3 concentration was observed (r = 0.75). Serum with low T3 AA concentration had apparent T3 concentration that was significantly (P < 0.01) lower than that in serum with high T3 AA concentration. Mean serum T4 concentration was not significantly different between serum with low or high T3 AA concentration. Mean Tg AA activity was significantly (P < 0.05) lower in serum with low T3 AA concentration than in serum with high T3 AA concentration. Addition of free T3 to serum significantly (P < 0.05) decreased detectable activity of Tg AA in both groups of sera. However, significant difference in magnitude of the reduction was not observed between sera with low or high T3 AA concentration. Results indicate that a fraction of Tg AA recognizes T3-containing epitopes in Tg. Increased prevalence of T3 AA for serum with high Tg AA activity indicates that T3 AA may be another valid indicator of lymphocytic thyroiditis. These antibodies may be generated against the hormonogenic epitopes of Tg.

Serum thyroid hormone concentrations and thyrotropin responsiveness in dogs with generalized dermatologic disease.

Fifty-eight dogs with generalized dermatologic disease that had not been given glucocorticoids systemically or topically within 6 weeks of entering the study were evaluated for thyroid function by use of the thyrotropin-response test. Dogs were classified as euthyroid or hypothyroid on the basis of test results and response to thyroid hormone replacement therapy. Baseline serum thyroxine (T4), free T4 (fT4), and triiodothyronine (T3) concentrations were evaluated in the 58 dogs. Serum T4, fT4, and T3 concentrations were evaluated in 200 healthy dogs to establish normal values. Hormone concentrations were considered low if they were less than the mean -2 SD of the values for control dogs. Specificity of T4 and fT4 concentrations was 100% in predicting hypothyroidism; none of the euthyroid dogs with generalized skin disease had baseline serum T4 or fT4 concentration in the low range. Sensitivity was better for fT4 (89%) than for T4 (44%) concentration. Significant difference was not observed in serum T4 and fT4 concentrations between euthyroid dogs with generalized skin disease and healthy control dogs without skin disease. Serum T3 concentration was not accurate in predicting thyroid function; most of the euthyroid and hypothyroid dogs with skin disease had serum T3 concentration within the normal range.

Comparison of two doses of aqueous bovine thyrotropin for thyroid function testing in dogs.

Thyroid function was evaluated in 20 healthy dogs by thyrotropin (TSH) response testing. Two dose regimens were used: 5 IU of TSH given IV and 1 IU of TSH given IV. Blood samples were collected prior to and at 4 and 6 hours after TSH administration. Serum was obtained and analyzed for total 3,5,3'-tri-iodothyronine and thyroxine (T4) concentrations by radioimmunoassay. All dogs were classified as euthyroid on the basis of response to 5 IU of TSH at 4 and 6 hours. The 1-IU dose of TSH failed to induce adequate increase in T4 concentration in 7 dogs at 4 and 6 hours when the criteria for normal response were post-TSH serum concentration T4 greater than or equal to 3.0 micrograms/dl and serum T4 increase by greater than or equal to 100% over baseline serum T4 concentration. One IU of TSH induced increase in serum T4 concentration over baseline; however, the increase was significantly (P less than 0.05) less than that in response to a 5-IU dose at 6 hours after administration of TSH.