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Conjunctival swabs from 44 free-living wild boars culled during a demographic control program applied in a Regional Park located in the Northern Italy were examined by 16S rRNA encoding gene nested PCR. In total, 22 (50%) wild boars were PCR positive. Sequencing of the amplicons identified Chlamydia suis and Chlamydia pecorum in 12 and 5 samples, respectively. For one sample found PCR positive, the nucleotide sequence could not be determined. Four conjunctival samples showed 92% sequence similarities to 16S rRNA sequences from Chlamydia-like organisms, as well as large intestine, uterus, and vaginal swab from the same four animals. Amoeba DNA was found in one Chlamydia-like organism positive conjunctival swab. To our knowledge, this is the first detection of members of the Parachlamydiaceae family in wild boars, confirming a large animal host range for Chlamydia-like organisms.

The aim of the present study was to evaluate the IgG and IgA seroprevalence to Simkania negevensis in adult patients with respiratory or gastrointestinal symptoms, by the microimmunofluorescence test (MIF). Serum samples were obtained fron 326 patients admitted to S. Orsola Hospital (Bologna, Italy) and collected a 24 month period in 2008-2010 for diagnostic investigations. As controls, 104 sera from blood donors were used. In subjects with gastrointestinal symptoms, we detected a S. negevensis IgG (68%) and IgA (18%) seroprevalence significantly higher than in the healthy controls. More extensive studies are needed to confirm the possible association of S. negevensis with gastrointestinal infections and its pathogen role.

Plasmids have been detected in the majority of strains in the genus Chlamydia and in many Chlamydophila species. Previous studies showed that FP Pring and FP Cello Chlamydophila felis strains have an extrachromosomial plasmid, whereas the FP Baker strain does not. Azuma et al. recently sequenced the entire genomic DNA sequence of the Japanese Cp. felis strain Fe/C-56 and described a 7,552 base pair circular plasmid. In the present study a highly conserved plasmid gene was detected in 11 Italian Cp. felis isolates, showing 100% nucleotide identity with the plasmid gene of Fe/C-56 Cp. felis strain.

Introduzione. Il Laboratorio di Microbiologia del Policlinico S. Orsola si occupa di diagnosi di infezioni da Chlamydia fin dal 1975: all’epoca non erano disponibili kit commerciali, pertanto le indagini microbiologiche prevedevano l’inoculo dei tamponi in colture cellulari e la colorazione del monostrato cellulare per evidenziare le inclusioni. Solo a metà degli anni ’80 cominciò la commercializzazione dei primi test immunoenzimatici per la ricerca di LPS e dei primi test molecolari, basati o sull’uso di sonde di ibridazione o su metodiche di amplificazione del DNA (LCR e PCR). Negli ultimi anni, infine, sono stati sviluppati nuovi metodi in Real-Time PCR per la determinazione contemporanea di DNA di C. trachomatis e Neisseria gonorrhoeae. Metodi. VERSANT® CT/GC DNA 1.0. Il test VERSANT® CT/GC DNA 1.0 (Siemens Healthcare Diagnostics) è basato su una procedura di PCR cinetica (kPCR) per la rilevazione del DNA di C. trachomatis e N. gonorrhoeae. IDEIA™ PCE Chlamydia. Il test IDEIA™ PCE Chlamydia (Oxoid) è un metodo immunoenzimatico per la determinazione di LPS di C. trachomatis. Risultati. Nel centro MTS del Policlinico S. Orsola sono stati eseguiti tamponi uretrali o cervicali. Dopo l’esecuzione, i pazienti sono stati invitati a procedere alla raccolta di un campione di urine. I tamponi sono stati analizzati con il metodo immunoenzimatico e solo il 5.0% di essi è risultato positivo. I campioni di urine sono stati studiati con il metodo molecolare e la percentuale di positività è aumentata a 11.9%. Conclusioni. La Real-Time PCR per la diagnosi delle infezioni da clamidia appare la metodica più sensibile e permette di analizzare campioni di urine, evitando in questo modo il ricorso a prelievi invasivi o dolorosi. Tale metodica sembra dunque particolarmente adatta per lo screening che, se applicato su larga scala, ci potrebbe dare una stima più realistica della prevalenza di questa infezione.

The modification of infectivity of different animal chlamydial species after their in vitro treatment with 10 and 80 µg/ml of PG-1 peptide, was studied. PG-1 at 10 µg/ml concentration had neither inhibitory nor increasing effects on the infectivity of all Chlamydia tested, whereas the incubation of 80 µg/ml of PG-1 with C. suis and Cp. pecorum isolates resulted in a twofold increase in the number of inclusions; a five- to tenfold increase in chlamydial infectivity was observed when Cp. psittaci and Cp. abortus strains were tested with the concentration of 80 µg/ml of PG-1 peptide. On the contrary, C. muridarum strain and Cp.felis isolates did not show any increase in the number of inclusions when incubated with PG-1 at a 80 µg/ml concentration. The different effect of PG-1 on the animals strains of chlamydiae could be due to the unique characteristics in the membrane structure of each strain.

Chlamydophila felis is a significant cause of conjunctivitis in cats in many countries. The transmission of chlamydial conjunctivitis from cats to human is possible, but a close contact is needed, so the risk for cat owners and people working in contact with cats is high. There have been very few reports in this connection, possibly because the transmission from cats to humans is a rare occurrence, or because such cases are underestimated as a result of failed isolation or inadequate investigation. In view of the C. felis circulation observed among feline populations, a seroepidemiological study was carried out about the circulation of C. felis in humans with and without contact with cats, using a microimmunofluorescence test (MIF). To verify cross-reactions with other species of chlamydiae, all the positive sera were tested by MIF also against C. psittaci and C. pneumoniae, and by immunoblotting analysis, to evaluate their reactivity to proteins of the three chlamydial species. A strong cross-reactivity between C. felis and C. pneumoniae was observed by MIF. By immunoblotting, all the C. felis-positive sera showed a strong reactivity with C. pneumoniae antigens, but a weak reactivity to C. felis and C. psittaci proteins. The results suggest no specific reactivity to C. felis in the sera examined. It is possible that, in the past, several cases of human diseases ascribed to C. felis infection on the basis of a not species-specific serology, were really due to C. pneumoniae. So, in the course of seroepidemiological surveys on feline chlamydiosis in the human population, cross-reactions between C. felis and C. pneumoniae should always be evaluated because of the higher seroprevalence of C. pneumoniae in the adult human population.

In this study the in vitro neutralizing activity of 71 swine sera resulted positive by microimmunofluorescence (MIF) test against Chlamydia suis was evaluated. These sera were collected from swine with clinical signs of conjunctivitis or reproductive disorders, reared in different farms in Emilia Romagna (Northern Italy). Neutralizing antibodies specific for C. suis were present in 57 out of the 71 swine sera tested (80.3%), positive by MIF with titres ranging between 32 and 2048. The 82.8% of the 71 swine sera tested presented by MIF titres ≥ 128. The presence of neutralizing antibodies was significantly correlated (p < 0.001) with MIF titres. All sera (11/11) positive by MIF with titres ranging from 512 and 2048 presented neutralizing antibodies and the 79.2% (42/53) of sera with MIF titres to C. suis ranging from 128 and 256 were neutralizing. Most of the sera presented a complement-dependent neutralizing activity even at a final dilution of 1:40.