H. Krauss

Role of Chlamydia trachomatis and Mycoplasmas in Chronic Prostatitis. A Review

Acute bacterial prostatitis caused by common urinary tract pathogens is an infrequent disease, and diagnostic difficulties are rarely encountered. On the other hand, chronic prostatitis is a common disease requiring rather elaborate diagnostic procedures. We applied the localization protocol of the four-specimen technique and combined quantitative determinations of microorganisms and quantitative cytologic analysis plus, in chlamydial infections, serologic investigations. Our studies provide good evidence that Ureaplasma urealyticum and Chlamydia trachomatis must be considered etiologic agents in many cases of chronic bacterial prostatitis. These unconventional microorganisms are assumed to infect the prostate by way of intracanalicular ascension from the urethra.

Monoclonal antibody based capture ELISA/ELIFA for detection of Coxiella burnetii in clinical specimens

A CAPTURE ELISA/ELIFA system based on monoclonal capture and biotinylated monoclonal detection antibody is described. The assay is fast, highly specific and detects a minimum dose of 2500 Coxiella (C.) burnetii particles. In contrast to the sophisticated and cumbersome isolation procedures, even non-specialized laboratories could use this assay system for investigating clinical samples of different origin for C. burnetii within a short period of time.

Analysis of the entire nucleotide sequence of the cryptic plasmid QpH1 fromCoxiella burnetii

The complete plasmid QpH1 fromCoxiella burnetii, isolate ‘Nine Mile’, phase I, was cloned asNotI fragment with a size of 37329 bp. The entire plasmid was sequenced by the chain termination method afterEcoRI subcloning. 37 open reading frames coding for polypeptides larger than 100 amino acid residues were determined. The predicted polypeptide products of the open reading frames were compared by computer analysis with reported protein sequences. Homologies of predicted polypetide products to analogous proteins are described.

Comparison of dark-field microscopy, culture, and polymerase chain reaction (PCR) for detection of Borrelia burgdorferi in field-collected Ixodes ricinus ticks.

In a study based on 100 field-collected female Ixodes (I.) ricinus ticks from the surroundings of Giessen, dark-field microscopy (DFM), culture, and PCR were compared as procedures for detecting Lyme borreliosis spirochetes in ticks. By DFM, 16 ticks were found to be infected with spirochetes. From the midgut of 18 ticks (including 14 microscopically positive specimens), spirochetes were cultured in BSK II medium and in BSK II medium supplemented with either co-trimoxazole (500 micrograms/ml) or 5-fluorouracil and kanamycin (200 micrograms/ml and 8 micrograms/ml). Using these selective media, the isolation rate was increased by 50% compared to BSK II medium without additives. Midgut homogenates of 22 ticks (including 13 ticks positive by culture and 12 microscopically positive ticks) were found to contain Borrelia (B.) burgdorferi-specific DNA by PCR using a primer set based on sequences of the flagellin gene of B. burgdorferi.

Studies of the prevalence of Coxiella burnetii, the agent of Q fever, in the foothills of the southern Bavarian Forest, Germany.

In studies carried out in 1991 in the foothills of the southern part of the Bavarian Forest, in the district of Freyung/Grafenau, ticks and small mammals were collected and examined for the presence of Coxiella (C.) burnetii and sera of small mammals and cattle investigated for antibodies against this rickettsia. A total of 1716 imagines and nymphs of Ixodes ricinus were collected by flagging and 892 larvae and nymphs of the same tick species removed from small mammals. In addition to 1095 serum samples from cattle, 326 specimens of nine species of small terrestrian mammals were examined. Neither in ticks nor in rodents, C. burnetii was detected, however, in 17 of 21 localities, seropositive cattle were found. Altogether, 12% of all 1095 heads of cattle tested were seropositive for C. burnetii antibodies. These serological results indicated a wide dissemination of C. burnetii in cattle of the region investigated, but there was no indication of a natural focus. As in other areas of Europe, an independent natural cycle of the agent involving cattle only is assumed to occur in this region.

The 16S/23S ribosomal spacer region ofCoxiella burnetii

The 16S/23S spacer region ofCoxiella burnetii isolate Nine Nile, phase 1, was sequenced. Sequence analysis revealed two tRNA coding regions for tRNAIle and tRNAAla. DNA sequence alignment demonstrated significant homology with tRNA species fromPseudomonas aeruginosa andRhodobacter sphaeroides, respectively. The non-coding tRNA spacer region was unique toCoxiella burnetii, based on database alignment.

Evaluation of the polymerase chain reaction (PCR) for detection of Chlamydia psittaci in abortion material from ewes

The polymerase chain reaction (PCR) was evaluated as a diagnostic tool for detection of Chlamydia (C.) psittaci in abortion material from 40 ewes. For this purpose, PCR results of 87 samples were compared with direct microscopic identification after chemical staining, cell culture isolation and a commercially available enzyme-linked immunosorbent assay (ELISA). The value for sensitivity as compared to cell culture was 97.7% whereas the specificity-value was calculated to be 84.1%.

Experimental Chlamydial Epididymitis

Male Wistar rats were infected with the chlamydial agent of guinea pig inclusion conjunctivitis by inoculation of chlamydiae into the vas deferens. Epididymitis was observed in all infected animals clinically and histologically. Chlamydiae were detected in the epithelium of epididymal tubules by immunohistochemical staining (alkaline phosphatase anti-alkaline phosphatase technique). Inflammation progressed from the cauda to the corpus and caput epididymidis leading to fibrosis of the cauda epididymidis 28 days after infection. Animals responded to the infection with a rise of both serum IgM and IgG antibodies.

Epidemiology and significance of Q fever in the Federal Republic of Germany

In the Federal Republic of Germany no large Q fever epidemics (more than 200 cases) have been encountered within the last 20 years; however, Q fever was prevalent throughout that period on a constant level (between 27 and 100 officially reported cases per year). Besides classical pneumonic Q fever, chronic forms associated with endocarditis, myocarditis and hepatitis were recently diagnosed for the first time in the Federal Republic of Germany. The disease Q fever in humans is often misdiagnosed as common cold or influenza, and more attention should be paid to this entity by the medical profession. Within ten years there has been a sharp increase of Q fever infections in livestock and pets as proved by seroepidemiologic investigations. Preliminary results of a seroepidemiological study indicate a parallel increase of seropositives in the human population, but further investigations on larger numbers of sera are required for statistic confirmation. There are reasons to believe that, in contrast to general opinion, in the Federal Republic of Germany C. burnetii is involved now in infertility in cattle, and besides being a zoonosis Q fever must be considered as a potentially important infectious disease of cattle causing economic losses in this country. Further investigations on this matter are required.

Analysis of caprine IgG1 and IgG2 subclass responses to Chlamydia psittaci infection and vaccination

Enzyme-linked immunosorbent assays (ELISAs) specific for caprine IgG(H+L), IgG1 and IgG2 were developed and evaluated for serodiagnosis of Chlamydia psittaci infections in a Tunisian goat flock with currently occurring chlamydial abortions and a clinically inapparent goat flock of an animal research facility. Additionally, ELISAs were applied to record the IgG1 and IgG2 dynamics of four goats vaccinated with inactivated Chlamydia psittaci and Coxiella burnetii. For screening purposes, the IgG(H+L) ELISA proved to be superior to the complement fixation test because it detected a larger number of chlamydial abortions and was easier to perform and to interpret. Analysis of Chlamydia psittaci-specific IgG1 and IgG2 responses to naturally occurring infections by ELISA revealed high IgG1 levels associated with IgG2 in goats with current abortions, whereas clinically inapparent, but seropositive goats were characterized by significantly lower IgG1 levels only (P less than 0.001). Similarly, the four vaccinated goats responded initially with Chlamydia psittaci-specific IgG1, whereas second and third vaccinations induced (as in goats with chlamydial abortions) predominantly IgG1, but also IgG2. The results indicated that clinically inapparent chlamydial infection may be distinguished from overt disease by analysing specific IgG1 and IgG2 responses. Applying Coxiella burnetii- specific ELISAs on field samples, IgG1 alone could be detected in eight, IgG2 alone in one and IgG1 combined with IgG2 in nine goats. The coxiella-specific antibody response of the four vaccinated goats was--in contrast to the chlamydia-specific response--characterized by IgG2 dominance.