Changes in the relative expression pattern of multiple vitellogenin genes in adult male and larval zebrafish exposed to exogenous estrogens.

Production of the lipoprotein vitellogenin (Vg) is induced in fish upon exposure to estrogens and is a biomarker of endocrine disruption in fish. In some fish, three types of Vg (VgA, VgB, and VgC) are recognized and transcribed from at least three distinct Vg genes (vtg). We investigated expression of vtg coding for Vg1A/B, Vg2A/B, and VgC in adult male and larval zebrafish exposed to various estrogenic substances. Quantitative PCR was conducted for transcripts of each vtg and a control gene (beta-actin). Male fish were exposed to 17beta-estradiol (E2) and 17alpha-ethinylestradiol, total RNA was extracted from excised liver, and histopathology of liver, trunk kidney, and gonads was conducted. Larval fish were exposed to 10 different estrogenic substances and total RNA was extracted from groups of whole larvae. In adult male fish, the relative fold change varied, but pattern of expression change (i.e., Vg1A/B > Vg2A/B > VgC) was consistent. Larger males exposed to E2 had significantly higher induction of each vtg. In larval zebrafish, the relative fold change in vtg expression varied according to specific estrogenic substance tested, but the pattern of change (i.e., Vg2A/B > Vg1A/B > VgC) was consistent for each substance that induced vtg.

Bordetella bronchiseptica fimbrial protein-enhanced immunogenicity of a Mannheimia haemolytica leukotoxin fragment.

Leukotoxin produced by Mannheimia (Pasteurella) haemolytica is an important virulence factor in shipping fever pneumonia in feedlot cattle and is a critical protective antigen. In this study, the immune response to a chimeric protein generated by combining a gene fragment encoding neutralizing epitopes of M. haemolytica leukotoxin and a fimbrial protein gene (fim N) from Bordetella bronchiseptica was evaluated. The recombinant gene was cloned in a bacterial expression vector under the control of the tac promoter and expressed as a fusion protein with glutathione-S-transferase (GST) in Escherichia coli. Immunization of mice with the recombinant protein, GST-LTXFIM elicited a significantly stronger anti-leukotoxin antibody response than comparable immunizations with GST-LTX fusion proteins lacking FIM N. The GST-LTXFIM was also more stable than GST-LTX during storage at -80 degrees C, thus alleviating a stability problem inherent to leukotoxin. This chimeric protein may be a candidate for inclusion in new generation vaccines against shipping fever pneumonia.

Clinical utility of measuring white blood cells on vaginal wet mount and endocervical gram stain for the prediction of chlamydial and gonococcal infections.

BACKGROUND: White blood cells on endocervical Gram stain and vaginal wet mount are frequently used to predict chlamydial and gonococcal infections. Previous studies provide conflicting evidence for the clinical utility of these tests. GOAL: To evaluate the clinical utility of measuring white blood cells on vaginal wet mount and endocervical Gram stain for the prediction of chlamydial infection and gonorrhea. STUDY DESIGN: Women undergoing pelvic examinations at 10 county health department family planning and sexually transmitted disease clinics were tested for chlamydial infection by ligase chain reaction assay (n = 4550) and for gonorrhea by culture (n = 4402). Vaginal wet mount and endocervical Gram stains were performed in county laboratories at the time of examination. RESULTS: The prevalences of chlamydial infection and gonorrhea were 8.8% and 3.2%, respectively. For detection of chlamydial or gonococcal infection, the likelihood ratio was 2.85 (95% CI, 2.10-3.87) for > 30 white blood cells on vaginal wet mount and 2.91 (95% CI, 2.07-4.09) for > 30 white blood cells on endocervical Gram stain. Similar results were seen for individual diagnoses either of chlamydial infection or of gonorrhea. CONCLUSION: Vaginal wet mount and endocervical Gram stain white blood cells are useful for the presumptive diagnosis of chlamydial infection or gonorrhea only in settings with a relatively high prevalence of infection or when other predictors can increase the likelihood of infection.

Selective screening for chlamydial infection: which criteria to use?

BACKGROUND: Screening sexually active women for Chlamydia trachomatis is necessary to detect asymptomatic infections. Selective screening is a common strategy because universal screening is too costly in many settings. In order to guide local programs in the choice of selective screening criteria, we examined the performance of previously proposed screening criteria for C. trachomatis. METHODS: A clinic-based, cross-sectional study was conducted in public family planning and sexually transmitted disease (STD) clinics in ten counties in North Carolina. Women (n = 4471 in family planning and n = 2201 in STD clinics) undergoing pelvic examination were enrolled consecutively. Nine sets of screening criteria, including age alone, were compared using sensitivity, specificity, number of tests required and receiver-operator characteristic (ROC) analysis. All women underwent testing with ligase chain reaction assay of cervical specimens to identify C trachomatis infection. RESULTS: The prevalence of C. trachomatis was 7.8% and 11.0% in family planning and STD clinics, respectively. The sensitivities of published criteria ranged from 0.50 to 0.97. Specificities ranged from 0.05 to 0.66. In family planning clinics, the best performing criteria would detect 84% of infections while screening 51% of women. In STD clinics, the same criteria would detect 83% of infections but require testing 67% of women. Testing women aged < or =22 would detect 77% of infections in family planning and 74% of infections in STD clinics, while testing 51% and 48% of the women, respectively. CONCLUSIONS: When site-specific criteria cannot be developed, age alone is an acceptable strategy for selective screening for chlamydial infection.

Coinfection with multiple tick-borne pathogens in a Walker Hound kennel in North Carolina.

Both dogs and humans can be coinfected with various Ehrlichia, Bartonella, Rickettsia, and Babesia species. We investigated a kennel of sick Walker Hounds and their owners in southeastern North Carolina for evidence of tick-borne infections and associated risk factors. A high degree of coinfection was documented in the dog population. Of the 27 dogs, 26 were seroreactive to an Ehrlichia sp., 16 to Babesia canis, and 25 to Bartonella vinsonii, and 22 seroconverted to Rickettsia rickettsii antigens. According to PCR results, 15 dogs were infected with Ehrlichia canis, 9 with Ehrlichia chaffeensis, 8 with Ehrlichia ewingii, 3 with Ehrlichia equi, 9 with Ehrlichia platys, 20 with a Rickettsia species, 16 with a Bartonella species, and 7 with B. canis. The detection of DNA from any Ehrlichia species was associated with clinical illness and with concurrent B. canis infection (by PCR). Both E. canis and an uncharacterized Rickettsia species appeared to result in chronic or recurrent infection. Death in the dog population was associated with living in a dirt lot rather than the concrete kennel. Of 23 people on whom serologic testing was conducted, eight were seroreactive to Bartonella henselae, one to E. chaffeensis, and one to R. rickettsii antigen; however, none had clinical or hematologic abnormalities consistent with illness caused by these organisms. We conclude that kennel dogs with heavy tick exposure can be infected at a high rate with multiple, potentially zoonotic, tick-borne pathogens. In addition, our findings further illustrate the utility of PCR for documenting coinfection with tick-transmitted pathogens.

Mild measles and secondary vaccine failure during a sustained outbreak in a highly vaccinated population.

A prolonged school-based outbreak of measles provided an opportunity to study "vaccine-modified" mild measles and secondary vaccine failure. Thirty-six (97%) of 37 unvaccinated patients had rash illnesses that met the Centers for Disease Control clinical case definition of measles, but 29 (15%) of 198 vaccinated patients did not, primarily because of low-grade or absent fever. Of 122 patients with seroconfirmed measles, 10 patients (all previously vaccinated) had no detectable measles-specific IgM and significantly milder illness than either vaccinated or unvaccinated patients with IgM-positive serum. Of 108 vaccinated patients with seroconfirmed measles, 17 patients (16%) had IgM-negative serology or rash illnesses that failed to meet the clinical case definition; their mean age (13 years), age at the time of vaccination, and time since vaccination did not differ from those of other vaccinated patients. The occurrence of secondary vaccine failure and vaccine-modified measles does not appear to be a major impediment to measles control in the United States but may lead to underreporting of measles cases and result in overestimation of vaccine efficacy in highly vaccinated populations.

Kawasaki syndrome: a controlled study of an outbreak in Wisconsin.

The etiology of Kawasaki syndrome remains unestablished, although a possible role has been suggested for exposure to the application of carpet shampoo, house dust mites, and rickettsial infection. During an outbreak of 20 cases of Kawasaki syndrome that occurred in southeastern Wisconsin from November 1982 through March 1983, a case-control study was done of 15 cases and 30 matched controls. The study included questionnaire administration, dust collection from homes, and serum specimen collection. Only one patient had been exposed to a shampooed carpet within 30 days before onset of illness. No differences were noted between cases and controls in the degree of exposure to house dust mite-associated factors in the home, nor in the occurrence, density and species-specific prevalence of house dust mites in the home. Meadow voles exposed to house dust mites from the homes of patients did not develop serologic or pathologic evidence of infection due to rickettsiae in the spotted fever and typhus groups or Coxiella burnetii. Anti-mite-specific immunoglobulin E was not detected in serum specimens from cases or controls. Results from this study do not support hypotheses suggesting that the development of Kawasaki syndrome is associated with exposure to application of carpet shampoo, house dust mites, or rickettsial infection.

Indirect fluorescent-antibody technique for serological diagnosis of La Crosse (California) virus infections.

A clinically relevant indirect fluorescent-antibody technique (IFA) was developed for the serological diagnosis of La Crosse virus infections. The IFA (67%) was as sensitive as the hemagglutination inhibition (58%) and neutralization (58%) tests in the detection of antibodies in acute-phase specimens. Immunoglobulin M antibodies were detected by the IFA test in 48% (11 of 23) of these specimens. Diagnostically significant increases in IFA titer were detected in 86% (19 of 22) of the paired samples. Antibodies were detectable in some patients 7 years after infection; however, the IFA test was not as sensitive as the other two tests in the detection of previous infections.