Survey of blood cultures methods in Italy in 2010

Sepsis is a serious clinical condition, associated with high mortality despite advanced modern medical treatment. Traditionally, the detection and identification of bacteria and fungi circulating in the blood-stream is based on blood cultures. A number of factors influence the yield of blood culture, most of them concerning the microbiologist skill and the laboratory organization. In order to collect information about the practices and procedures used for the detection of microrganisms in blood cultures in the italian laboratory (lab), an e-mail with the invitation to participate in the survey was sent to 2000 members of the Italian Association of Clinical Microbiology. Responses were received from 100 lab, located from all over the country (in 18/20 italian regions). The results presented hereby concern specimen collection, culture techniques, rapid identification and susceptibility testing, laboratory organization, relationships with physicians. In summary, most lab use automated systems (96%), the bottles are incubated immediately during public holidays in 72/96 lab (75%) and in 49/97 lab at night (50.5%), the lenght of incubation was 5 or 7 days in 93% of the lab, although it is common to extend the incubation period when brucellosis (74 lab), endocarditis (49 lab), systemic mycosis (33 lab) is suspected. A wide variety of media are employed for subcultures. All lab process the positive bottles at least once a day, while only in 42 of 81 (51.9%) lab the positive blood are processed on holiday. Communication between clinicians and microbiologist include: distribution of specimen collection guidelines (96/100 lab), availability to microbiologist of patients’ clinical situation (77/96 lab, 80.2%), and adding to report the microbiologist’ suggestion (75/98 lab, 76.5%). The results, compared with those collected with a similar questionnaire in 2001, show a greater adherence to guidelines: the number of bottles examined by lab yearly is almost doubled, the length of incubation is shortened to 5 days in 42% (vs 9.2% in 2001), direct susceptibility tests seem to be performed more frequently (in 29% of lab vs18.6% in 2001, mostly in larger hospitals), more lab process positive bottles on Sunday, cooperation with clinicians is improved

Epidemiology of intestinal parasitosis in Italy between 2005 and 2008: diagnostic techniques and methodologies

Aim of the study was to keep a real and actual photo relating to 2005-2008 regarding to diagnostic techniques and methodologies for intestinal parasites; so it would be possible to know specific epidemiology and suggest more rational and efficacious guide-lines. All members of AMCLI were involved in the proposal of a retrospective study regarding bowel parasites, helminths and protozoa.To engaged laboratories we asked how O&P was performed, if a specifical research for E. vermicularis and S. stercoralis was performed, if for the identification of D. fragilis, Entamoeba histolytica/dispar and Cryptosporidum spp were performed recommended specific permanent stains. 23 laboratories gave assent; but for an inferior number was possible to use the data for analysis and evaluation. Relating O&P only some laboratories performed permanent stains: Giemsa for D. fragilis, antigen and/or Trichrome stain for E. histolytica/dispar, antigen and/or acid fast stain for Cryptosporidium spp.Not all laboratories research specifically S. stercoralis. So the epidemiology is differentiated and related more to adequate or not adequate techniques than cohorts of examined populations. The overall positivity for parasites ranged from 0% to18.7%,for protozoa (pathogens or not) were from 0% to 14.7%; for nematodes from 0% to 3.7%; for cestodes from 0% to 1.0%; for trematodes from 0% to 1.0%.Among helminths, E. vermicularis, followed by S. stercoralis, also in O&P, is the most frequent.The specific research of S. stercoralis gave a positivity from 0% to 33.3%; the cellophane tape test was positive for E. vermicularis from 0% fo 21.9% of cases.Among pathogen protozoa, D. fragilis, when permanent stain were applied, prevailed from 0% to 16.6%; G. duodenalis from 0.8% to 4.3%; E. histolytica/dispar, using a permanent stain or research of antigen, was identified from 0% to 20.6%. Coccidia were very rare, with Cryptosporidium spp observed from 0% to 5.2%. These are our conclusions: many laboratories need for a critical revision of own technical procedures and diagnostic methodologies

Nuovo terreno cromogeno MRSA ID per la rilevazione di S. aureus meticillino-resistente: affidabilità nella lettura

We proceeded to the evaluation of the reability of reading and interpretation of cultures on the chromogenic medium MRSA ID for the detection of methicillin-resistant S. aureus, within a surveillance program. Our observations demonstrate such method results a useful instrument of screening for the prevention and the control of the spread of the infections by MRSA

The State of Microbiology Diagnostic of Prosthetic Joint Infection in Europe: An In-Depth Survey Among Clinical Microbiologists

Background: This study aims to give an overview on how microbiology diagnosis tests of Prosthetic joint infections (PJI) is performed in Europe, and to explore whether any factor influences the decision on implementing a test. Methods: An extensive online survey of clinical microbiologists from seven European countries (Belgium, Estonia, Germany, Italy, Netherlands, Switzerland, and Spain). Following items were assessed: (i). general information on the laboratory, (ii) preference of the laboratory and clinical microbiologists regarding samples, (iii) transportation and (iv) processing of explanted foreign bodies and tissues and synovial fluid, (v) culture media and culture duration, (vi) reporting (identification and susceptibility testing), and (vii) use of molecular microbiology techniques. Results: Invited were 163 clinical microbiologists. The response rate from each country was above 50% (range 51–78%), except for Germany (36%). Frequent PJI diagnostics were the use of tissue pre-processing (58.1%), culturing synovial fluid in blood culture bottles (45.5%), use of sonication for processing explanted prosthesis (56.8%), reporting the presence of synovial leukocyte counts (67%), use of blood aerobic and anaerobic agar (97.7%), and enrichment media thioglycolate (69.3%). The most common incubation time of the culture media is 7–14 days (34.1–70.5%). The clinicians were called to report the culture results (80.7%), and to give antibiotic recommendation (67%). Conclusion: There are common practices in processing PJI samples and reporting results, which is promising for harmonization of PJI diagnostic in the future. However, variation in diagnostic tests should also be considered in interpreting and comparing clinical microbiology results

Prevalence of Diphyllobothrium latum (Cestoda: Diphyllobothriidae) plerocercoids in fish species from four Italian lakes and risk for the consumers

In recent years there has been a re-emergence of diphyllobothriasis by Diphyllobothrium latum (Cestoda: Diphyllobothriidae) in Italy, France and Switzerland, where in the past this fish-borne zoonosis was widespread and then virtually disappeared. A change in eating habits such as the consumption of raw/undercooked freshwater fish, has led to an increased risk for consumers of ingesting infective larvae of D. latum. A survey on the factors responsible for the re-emergence of human diphyllobothriasis in Italy was carried out from March 2013 to December 2014. The aim of this study was to assess the diffusion of D. latum plerocercoids in the fish populations of the sub-alpine lakes of Maggiore, Como, Iseo and Garda, updating the scarce historical data and assessing a preliminary \u201crisk level\u201d of the lacustrine environments and fish species under investigation. A total of 2228 fish belonging to 5 species, 690 from Lake Maggiore, 500 from Lake Como, 655 from Lake Iseo and 383 from Lake Garda were submitted to parasitological examination. The presence of D. latum plerocercoid larvae was detected in 6.6%, 25.4% and 7.6% of perch (Perca fluviatilis) from Lakes Maggiore, Como and Iseo respectively. The parasite was also present in pike (Esox lucius) with prevalence values ranging from 71.4 to 84.2% and in 3.6\u20133.8% of burbot (Lota lota) from Lakes Iseo and Como. Fish from Lake Garda were negative as well as sampled whitefish (Coregonus lavaretus) and shad (Alosa fallax lacustris). The results of this survey showed a widespread presence of D. latum plerocercoid larvae in Maggiore, Como and Iseo fish populations. Urban fecal contamination of water is still a key issue to be resolved, together with the improvement of communication with consumers regarding the best dietary habits and the most effective processes of parasite inactivation, required for the consumption of raw/undercooked fish caught in high-risk areas

Microbiological characterization of plasmid-mediated AmpC ß-lactamases and E. coli hyperproducers: how and why ?

The aim of this study is the evaluation of phenotypic method for the detection of plasmid-mediated AmpC producing Enterobacteriaceae by agar diffusion.We developed a phenotypic method with double disk test (CLSI) and evaluation of synergism between Cloxacillin and/or Boronic Acid with cefotaxime and ceftazidime and cefepime with amoxicillin/clavulanic acid. As reference method for AmpC detection we used a multiplex PCR according to Perez-Perez. Among 7476 Enterobacteriaceae we detected 45 strains: 37 (82.2%) plasmid-mediated AmpC producers, 6 (13.3%) E. coli hyperproducers and 2 E. coli (4.5%) positive for both.The AmpC phenotypic test was positive for all the isolates, showing a typical ghost zone between cloxacillin and cephalosporins or boronic acid and cephalosporins.The AmpC multiplex PCR confirmed that 28 P. mirabilis and 7 E. coli harboured a gene belonging to the bla-CMY-LAT family. Sequencing defined the presence of CMY-16 in all P. mirabilis, CMY-2 in E. coli, DHA-1 in 3 K. pneumoniae and FOX in 1 K. pneumoniae and allowed us to identify eight strains as E. coli hyperproducer: six E. coli yielded no amplicon and 2 were also producer of CMY-2. In this study the phenotypic method showed a sensitivity and a specificity of 100%.Waiting for the indication of international authorities, we think this phenotypic screening method could be useful in the routine of microbiological laboratories

Antimicrobial susceptibility of enterococci from Italian patients with infective endocarditis

The epidemiological and clinical study “Studio Endocarditi Italiane” (SEI) evaluated 947 cases of infective endocarditis in 2004-2008. Enterococcal aethiology was described in 11% cases.The aim of our study was to evacuate the in vitro activity of twelve antibiotics alone and in association against 20 strains (15 E. faecalis, 5 E. faecium). Results show the in vitro activity of tigecyicline, daptomicine and linezolid. Synergism is appreciated for ceftriaxone-fosfomycin (FICindex50=0.34, FICindex90=0.56) against E. faecalis and for imipenem-fosfomycin against E. faecium.The majority of the tested strains show addition using ampicillin-ceftriaxone (FICindex50=0.85, FICindex90=1.01) and moxifloxacin- tigecycline (FICindex50=0.77, FICindex90=1.96)

Typing of Enterococcus spp. strains isolated from patients with infective endocarditis by an automated repetitive-sequence-based PCR system

The use of rep-PCR was studied to characterize 32 isolates of Enterococcus spp. from Italian patients with infective endocarditis and determine specific banding patterns associated with antimicrobial resistance phenotypes. The results obtained with rep-PCR were analyzed with respect to the antimicrobial profile by a MIC evaluation (E-test) of all strains tested. The automated rep-PCR technique resulted an easy method for the characterization of Enterococcus spp. It confirmed the eterogenicity of enterococcal strains involved in infective endocarditis. Furthermore results of antimicrobial susceptibility revealed that rep-PCR can not be used as diagnostic tool to characterize and identify difference between resistant and susceptible Enterococcus spp. strains