Sex abstinence for at least three days before sampling. No antibiotic for at least 1 week before sampling.
Bugs present in the whole genital tract:
To better differentiate prostatic germs:
All these samples will be analysed for the presence of all common bacteria, yeasts, urogenital mycoplasmata, DNA of Chlamydia trachomatis, Neisseria gonorrhoeae, HPV, and additionally for specific Chlamydia t. secretory IgA, that represents a good local marker of infection. The results will also be confirmed by Electron Microscopy.
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S. Mazzoli,1 A. Salvi,1 F. Guercini,2 M. Marongiu,1 A. Raugei 3 1 Centro MST, Ospedale S.M. Annunziata, Firenze, Italy 2 Istituto di Urologia, Universita’ degli Studi, Perugia, Italy 3 Istituto di Urologia, Universita’ degli Studi, Firenze, Italy
Chlamydia pneumoniae (C.p.) is one of the newest pathogens of the respiratory tract in humans. Every year almost 10% communicable pneumonitis are caused by this microorganism. The seroprevalence of C.p. in normal populations is high, estimated to be 50% at the age of fifthly, confirming its wide diffusion. Recently, C.p. has been connected with coronary chronic disease and myocardial infarction. Very recently C.p. has been found in patients with interstitial cystitis, a condition related to prostatitis.
We have analyzed for the presence of C.p.DNA, by nested PCR, prostatic biopsies, EPS, post EPS urine, total ejaculate and first void early morning urine from patients affected by different prostatic pathologies: chronic abacterial prostatitis, benign prostatic hyper-plasia (BPH) and prostate cancer. 40 patients were included in the study and 87% resulted positive for Chlamydia pneumoniae DNA. 100% of the prostate biopsies (N. 10 patients) were positive, demonstrating the presence of the microorganism inside the prostate gland both in prostatitis, BPH and prostate cancer patients.
Chlamydia pneumoniae, a microorganism inducing chronic body damages, has to be better studied in relation to chronic prostatic pathologies and prostate cancer. Several interrogatives remain also open: the role of machrophages and other immunologically related cells in transporting the microrganism inside the prostate gland and in modulating the infection;
its persistence in relation to the various stages of prostate damage.
C.p. positivity in these chronic prostatitis, resistant to several therapeutic regimens of antibiotics, open new pharmacological approaches. The constant presence of Chlamydia pneumoniae in all the prostate pathologies examined open a discussion about the role of this microrganism in their development during the time: we postulate that the three conditions - prostatitis, BPH, prostate cancer - may represent different moment of the same process in which external conditions due to the host, especially immunological conditions, can induce the determinism of the one instead of the other pathology.
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S Mazzoli, A Salvi, C Romeo, l Ramacciotti STD Center, Santa Maria Annunziata Hospital, SHA of Florence, Florence, Italy
Chronic prostatitis represents one of the emerging infective problems in young male adults of our area in Italy. Mean age in these patients is confirmed around 30-31 years. Due to early advent of symptoms and the fact that symptoms persist for years, considering the relative young age of these patients and additionally the fact that no common bacteria, fungi, or mycoplasmata were found by conventional culture methods in semen or /and prostatic secretions, we started to look for other micro-organisms, difficult to culture, that the patient could acquire in young age, probably together with the beginning of sexual activity.
With the introduction in Italy of molecular diagnostic methods to detect bacterial and viral DNA (PCR), we started to search systematically STDs agents such as Chlamydia trachomatis (Ct), Neisseria gonhorroeae (Ng) (Roche Molecular System USA), and, later in 1998, Human Papilloma Virus (HPV) (Diatech-ltaly, Alphagenics Diaco Biotecnology ltaly, Bioline Diagnostics-ltaly, Cytic USA) in chronic prostatitis patients" biological fluids. Out of 820 chronic abacterial prostatitis patients screened for Ct and Ng DNA in semen in the last 2 years, 15.7% (129 patients) tested positive; prevalence rate for single micro-organism was 7.80% and 7.92%, respectively. None of them tested positive in urethra. Out of the 114 patients screened for presence of HPV DNA in semen, 30.70 % were positive. The co-infection rate for Ct/HPV was 51.4% and for Ng/HPV it was 34.2; 82.35% of Chlamydia t. and/or Gonococcus DNApositive patients were ems to be underestimated if we consider the 40.2 % Western blot confirmed specific anti Chlamydia t. secretory IgA positives in semen.
Detecting DNA, we proved microorganism presence in 20% (165/820) abacterial prostatitis patients. The high co- infection rate between these STDs agents emphasises about common risk factors strictly connected with infections by these microorganisms early, probably at the start of sexual activity.
In a recent statistical study from our Laboratory data, I have found out that Ureaplasma urelyticum represents the most common bacterial isolate from total ejaculate of our prostatitis patients: 172 U. u. strains out of 933 different microrganisms isolated, that means 18,43%, that's a high prevalence rate. In addition in all these patients the bacterial presence was 10,000 Colony Forming Units, that's means infection. Our sampling protocol exclude lower genital tract contamination of total ejaculate.Antibiotic susceptibility is under evaluation and as soon as possible I will inform you of the in vitro resistance and susceptibility rates (%). But always in our experience I can say that what we see in "in vitro studies" it is not exactly what it works in vivo!
Co-infected by HPV, either low or high risk HPV strains. Gonococcai and Chlamydial co-infection rate was 9.3%, but it seems Chlamydia trachomatis is one of the most prevalent microorganism in non-bacterial prostatitis patients in my area in Italy; I can say that Chlamydia, like Ureaplasma urealyticum, in my experience, is unfortunalely really difficult to eradicate. We perform very long term follow up in our prostatitis patients with chlamydial infections by DNA detection and specific chlamydial immunological markers, and, recently, we have found intra-macrophages Chlamydial bodies after repetitive cycles of tetracycline theraphy in some of our patients. So, we have the evidence that Chlamydia is mantained inside immunocompetent cells in a state able to re-induce possible infection in permissive conditions. This is an experimental evidence in our chronic Chalmydial prostatitis patients. So that we have to go on with further studies. Of course this is a very serious problem!! It seems we are in front of a very "smart" organism!!
F.Guercini1, S.Mazzoli,2 C.Pajoncini,1 A.Salvi,2 M.Porena 1,
1Dept. of Urology, University of Perugia, Italy. 2STD Centre, St. M. Annunziata Hospital, Florence, Italy
Objectives: In the National Institute of Health Classification prostatitis syndrome are categorized in clinical practice on the basis of bacteriuria and the number of inflammatory cells in the prostatic secretion. According to these indications many patients are considered carriers of abacterial prostatitis and are treated as such. The presence of an agent inflecting the prostate cannot however be completely excluded. It could nest in acini or in fibrous-calcifications but still be active and capable of rendering the socalled abacterial prostatitis chronic.
Materials & Methods: We recruited 56 of the last 145 patients referred to our Centre because of prostatitis. The age-range was 18-46 years (mean 32 years). No patient was affected by bacteriuria. In the prostatic secretion all had at least 10 leucocytes per microscopic field according to Stamey’s method (1966). 2-4 weeks before prostatic secretion sampling, urethral swabs showed no patient was positive for Trichomonas vaginalis, Chlamydia tracomatis, Mycoplasma hominis, Ureaplasma urealyticum, HPV ed Herpes genitalis. Using the transper-ineal route, ultrasound guided needle aspirates were taken from sonographically dishomogenous areas in the prostate. Samples underwent histological analysis and DNA extraction to detect Chlamydia trac-omatis, Neisseria gonorrhoeae and HPV using PCR amplification.
Results: Histological findings were indicative of inflammation in all 56 patients with lymphocyte aggregates being found rarely within the gland (19%) and mostly in the perigland area (46%) and stroma (35%). Cultures were positive for aerobic (56%) and anaerobic (23%) agents. Twentyone patients (38%) presented with more than 2 species of microorganisms and 9 (15%) with more than 3. DNA infected with Chlamydia trachomatis was found in 19 (34%) patients. The 9 samples (16%) with only anaerobic bacteria were associated with a high number of leucocytes in Stamey’s test (>15).
Conclusions: The accuracy of needle sampling under ultrasound guidance using the transperitoneal route excludes false positive results caused by contamination with pathogens in the urethra. The high frequency of positivity for microorganisms detected using these techniques indicates studies on more patients should be performed in order to revise the classification of prostatic syndromes and to define them more accurately.
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