Extended-spectrum ß-lactamase, AmpC-producing, and fluoroquinolone-resistant Escherichia coli in retail broiler chicken meat, Italy

Background: Globally, antimicrobial drug-resistant Escherichia coli is among the most common etiological agents of invasive disease in humans. In Europe, increasing proportions of infections due to third-generation cephalosporins and/or fluoroquinolone-resistant extraintestinal pathogenic E. coli (ExPEC) strains are reported. E. coli from poultry are those more closely linked to human E. coli, but lack of reliable data makes it difficult to assess the attributable risk of different food sources. In the present study, our objective was to investigate the antimicrobial resistance profile, phylogenetic background, and virulence factors of E. coli isolates from broiler chicken meat sold at retail in Palermo, Italy. Materials and Methods: Isolation of multidrug resistant (MDR) E. coli was performed during April-December 2013 on a total of 163 chicken meat samples. Susceptibility to a panel of nine antimicrobial agents was determined. PCR assays were carried out to detect extended-spectrum β-lactamase (ESBL), plasmid-mediated AmpC β-lactamase, and plasmid-mediated quinolone resistance (PMQR) genes, phylogenetic group, and ExPEC-associated traits. A single nucleotide polymorphism (SNP) PCR was done to detect E. coli sequence type (ST)131. Results: One hundred thirty-four isolates from 109 meat samples were MDR. B1 was the most prevalent phylogenetic group (47.8%), followed by groups D (25.4%), A (22.3%), and B2 (4.5%). ESBLs and AmpC β-lactamases were detected by PCR in 132 (98.5%) and 15 (11.2%) isolates. PMQR determinants were detected in 122 (91%) isolates. Twenty-two MDR isolates met the molecular definition of ExPEC. SNP-PCR results confirmed that four B2 isolates were ST131. Enterobacterial Repetitive Intergenic Consensus sequence-PCR analysis showed a large heterogeneity with 55 unique profiles and 31 clusters including 2-4 isolates. Conclusions: An alarmingly high prevalence of MDR E. coli from broiler chicken meat is evident in our geographic area. The ongoing use of antimicrobial drugs in livestock should be urgently restricted, particularly in the poultry sector

La sorveglianza della tubercolosi in Sicilia

Obiettivi. Descrivere l’andamento dei casi di tubercolosi (TB) in Sicilia negli anni 2010-2013. Disegno e partecipanti. Indagine osservazionale delle notifiche di TB relative agli anni 2010-2013 mediante l’analisi dei dati del Sistema informativo delle malattie infettive (SIMI) e indagine epidemiologico-molecolare mediante tipizzazione spoligo- e 24- loci mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) dei ceppi di Mycobacterium tuberculosis complex (MTBC) isolati a Palermo negli anni 2012-2013. Risultati. Negli anni 2010-2013 sono stati notificati in Sicilia 876 casi di TB con un tasso medio annuale di notifica pari a 4,4 casi per 100.000 abitanti. La proporzione di casi di TB in soggetti nati all’estero è crescente e nel 2012 e 2013 supera quella nella popolazione autoctona. L’età mediana dei pazienti stranieri è risultata significativamente inferiore agli italiani (30,5 vs. 51,2 anni, p <0,001). La distribuzione per classi di età è stata significativamente differente nelle due subpopolazioni, con la proporzione più elevata di casi nelle fascia di età 15-44 anni tra gli stranieri e, al contrario, una distribuzione uniforme dei pazienti italiani tra le fasce di età 15-44, 45-64 e >64 anni. Nell’82,5% dei casi è stata diagnosticata una localizzazione polmonare. La tipizzazione molecolare è stata effettuata su 151 ceppi di MTBC. Mediante spoligotyping essi sono stati attribuiti a 14 differenti lineage e 33 sublineage rappresentati con frequenza diversa tra le due subpopolazioni di pazienti. Soltanto nove ceppi (6,2%) sono stati inclusi in quattro cluster. Sono stati identificati, infine, due ceppi multiresistenti. Conclusioni. L’epidemiologia della TB in Sicilia è complessa e in rapida evoluzione, con una crescente proporzione di casi in soggetti non italiani e una grande eterogeneità dei ceppi di MTBC circolanti. L’integrazione di strumenti convenzionali e molecolari è necessaria per una sorveglianza efficace e un’interpretazione accurata dell’epidemiologia della TB in Sicilia

Extraintestinal pathogenic Escherichia coli sequence type 131 H30-R and H30-Rx subclones in retail chicken meat, Italy

Extraintestinal pathogenic Escherichia coli sequence type 131 (ST131), typically fluoroquinolone-resistant (FQ-R) and/or extended-spectrum β-lactamase (ESBL)-producing, has emerged globally. Among clinical isolates, ST131, primarily its H30-R and H30-Rx subclones, accounts for most antimicrobial-resistant E. coli and is the dominant E. coli strain worldwide. We assessed its prevalence and characteristics among raw chicken meat samples on sale in Palermo, Italy. A collection of 237 fluoroquinolone resistant and ESBL/AmpC producing E. coli isolates, which had been isolated from processed retail chicken meat in the period May 2013-April 2015, was analyzed. Established polymerase chain reaction methods were used to define ST131 and its H30 subclones, ESBL, AmpC, and plasmid-mediated quinolone resistance (PMQR) determinants. Amplified Fragment Length Polymorphism (AFLP) was performed to assess the relatedness among ST131 isolates. Out of the 237 E. coli isolates, 12 isolates belonged to the phylogenetic group B2. Based on the molecular definition of ExPEC, all isolates were attributed with the status of ExPEC. SNP-PCR results confirmed that nine isolates were ST131. SNP-PCR for H30-R and H30-Rx subclones showed that six and three ExPEC ST131 were positive for H30-R and H30-Rx, respectively. The results of AFLP showed that, except for four isolates grouped into two clusters which proved to be indistinguishable, the isolates under study were genetically heterogeneous. To the best of our knowledge, this is the first report of H30-R and H30-Rx subclones in animal food samples. Our findings appear to support the role of food chain in their transmission to humans

NDM-1 and OXA-163 producing Klebsiella pneumoniae isolates in Cairo, Egypt, 2012

Here we describe carbapenem resistance determinants in two Klebsiella pneumoniae isolates recovered from two hospitalised patients in the same intensive care unit of a cancer hospital in Cairo, Egypt. PCR and sequencing were used to detect and characterise β-lactamase genes. Clonal relationships between the isolates were analysed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The first K. pneumoniae isolate carried the blaNDM-1 gene and the second isolate carried the blaOXA-163 gene. Both isolates co-expressed the extended-spectrum β-lactamase CTX-M-15. The two isolates belonged to different sequence types (STs), ST11 and ST16, respectively. No history of travel was established for the two patients. The first identification of NDM-1-producing K. pneumoniae in Egypt adds further evidence to the spread of NDM-1-producing Gram-negative micro-organisms in North Africa. The additional detection of blaOXA-163 in a K. pneumoniae isolate confirms its endemic presence in a critical healthcare setting of this geographic area

Cephalosporin resistant Escherichia coli from cancer patients in Cairo, Egypt.

Cephalosporin-resistant Escherichia coli has been increasingly reported worldwide. In this study, 32 cephalosporin resistant E. coli isolates identified from cancer patients in Cairo, Egypt in 2009-2010 were analyzed. Twenty-three were of phylogenetic group D, seven A and one each B1 and B2. By rep-PCR 15 phylogroup D isolates were grouped in four clusters, one with sequence type (ST) 405 and three ST68. Seventeen isolates showed single patterns. blaCTX-M-15 and aac(6')-Ib-cr were the most common resistance determinants. blaOXA-48 and blaVIM were also detected. Multidrug resistant E. coli seriously affects healthcare, especially in immunocompromised hosts, such as cancer patients

Enhanced surveillance of invasive listeriosis in the Lombardy region, Italy, in the years 2006-2010 reveals major clones and an increase in serotype 1/2a

BACKGROUND: Invasive listeriosis is a rare, life-threatening foodborne disease. Lombardy, an Italian region accounting for 16% of the total population, reported 55% of all listeriosis cases in the years 2006-2010. The aim of our study was to provide a snapshot of listeriosis epidemiology in this region after the implementation of a voluntary laboratory-based surveillance system. METHODS: We characterized by serotyping, pulsed-field gel electrophoresis, multilocus sequence typing and detection of epidemic clone markers, 134 isolates from 132 listeriosis cases, including 15 pregnancy-related cases, occurring in the years 2006-2010 in Lombardy. Demographic and clinical characteristics of cases have also been described. RESULTS: The mean age of non pregnancy-associated cases was 64.7 years, with 55.9% of cases being older than 65 years. Cases having no underlying medical conditions accounted for 11.6%. The all-cause fatality rate of 83 cases with a known survival outcome was 25.3%. Serotypes 1/2a and 4b comprised 52.2% and 38.8% of isolates, respectively. Seventy-three AscI pulsotypes and 25 sequence types assigned to 23 clonal complexes were recognized. Moreover, 53 (39.5%) isolates tested positive for the epidemic clone markers. Twelve molecular subtype clusters including at least three isolates were detected, with cluster 11 (1/2a/ST38) including 31 isolates identified during the entire study period. No outbreaks were notified to public health authorities during this period. CONCLUSIONS: The findings of our study proved that epidemiology of listeriosis in Lombardy is characterized by a high prevalence of major clones and the increasing role of serotype 1/2a. Molecular subtyping is an essential tool in the epidemiology and surveillance of listeriosis. Rapid molecular cluster detection could alert about putative outbreaks, thus increasing the chance of detecting and inactivating routes of transmission

Healthcare associated pathogens in a changing world

In developed countries about 10% of the hospitalizations are complicated by a healthcare-associated infection [1]. Up to 75% of these infections are due to multidrug-resistant organisms (MDROs) [1]. Antimicrobial resistant bacterial infections are associated to higher morbidity, mortality and healthcare costs than those caused by susceptible organisms [1]. The findings of the point prevalence survey in European acute care hospitals published in 2013 by the European Centre for Disease Control and Prevention (ECDC) show large variations between countries and between different regions of the same country, with Italy being allocated within the high-endemic areas for both MRSA and MDROs [2]. Despite antimicrobial resistance affects most bacterial species, MDR Gram negatives represent the most serious threat. In a few years Enterobacteriaceae, mainly Escherichia coli and Klebsiella pneumoniae, have evolved from extended spectrum β-lactamase (ESBL) producing to carbapanem-resistant organisms [3]. Simultaneously, Acinetobacter baumannii has quickly become extremely or pan-drug resistant [4]. Carbapenem resistant Gram negatives heavily impact on clinical outcomes with mortality rates significantly higher than the susceptible strains of the same species [1]. Of further concern, very few antimicrobial agents are available for an effective treatment of these infections and new agents active against these organisms are not currently in development. Many intertwining factors are driving these epidemiological changes, involving patients, healthcare delivery systems, infection control practices and, most important, misuse and inappropriate use of antibiotics in all healthcare facilities, in community and in animal husbandry. In particular, the transition of the healthcare delivery systems from a hospital-centered model to a healthcare facility network has gradually blurred the borders between hospital and community and the patients’ travel within this network has critically contributed to disseminate MDROs [5]. As a consequence, antimicrobial resistance is now as common, if not more so, in post-acute clinical facilities, such as long term care settings and nursing homes [5]. The “revolving door” is the very efficacious image used as the paradygm of the spreading routes of organisms with hospital and community reservoirs, as E. coli or MRSA. The revolving door, indeed, enlightens how the colonized patients entering back and forth several healthcare settings drive the amplification of the antibiotic resistance [6]. Stringent infection control and prevention practices and wise use of antibiotics are unanimously agreed as the key actions to fight MDROs. Of course, we need new antibiotics, but first we have to learn how to protect them from a precipitous erosion of their effectiveness

Methicillin-resistant Staphylococcus aureus nasal colonization in a level III neonatal intensive care unit: Incidence and risk factors

Objective: To describe epidemiologic features and identify risk factors for methicillin-resistant Staphylococcus aureus (MRSA) acquisition in a level III neonatal intensive care unit (NICU). Setting: A prospective, cohort study in a university-affiliated NICU with an infection control program including weekly nasal cultures of all neonates. Methods: Demographic, clinical, and microbiologic data were prospectively collected between June 2009 and June 2013. Molecular characterization of MRSA isolates was done by multilocus variable number tandem repeat fingerprinting, staphylococcal cassette chromosome mec typing, and on representative isolates by multilocus sequence typing and s. pa typing. Results: Of 949 neonates, 217 (22.87%) had a culture growing MRSA, including 117 neonates testing positive at their first sampling. Of these latter infants, 96 (82.05%) were born with the infection and 59 (50.43%) had been transferred from the nursery. Length of stay and colonization pressure were strong independent predictors of MRSA acquisition. Among MRSA isolates, 7 sequence types were identified, with ST22-IVa, spa type t223, being the predominant strain. Conclusions: In an endemic area, early MRSA acquisition and high colonization pressure, likely related to an influx of colonized infants from a well-infant nursery, can support persistence of MRSA in NICUs. Surveillance, molecular tracking of strains, and reinforcement of infection control practices, involving well-infant nurseries in a comprehensive infection control program, could be helpful in containing MRSA transmission