Description of novel resistance islands harbouring blaCTX-M-2 in IncC type 2 plasmids.

OBJECTIVES: We sequenced two IncA/C plasmids harbouring blaCTX-M-2 in Klebsiella pneumoniae clinical isolates and compared their antibiotic resistance islands. METHODS: Transconjugants were obtained from two clinical K. pneumoniae isolates harbouring blaCTX-M-2. Plasmid DNA from transconjugants underwent short-read whole-genome sequencing, reads were assembled, and gaps were closed by PCR and sequencing. Determination of plasmid replicons, antibiotic resistance genes, identification and characterisation of insertion sequence (IS) elements, and comparison with publicly available plasmid sequences were performed. RESULTS: blaCTX-M-2 was located in a complex class 1 integron In35::ISCR1::blaCTX-M-2, inserted in two different transposons designated Tn7057 and Tn7058, that reside in the resistance islands of plasmids pUR-KP0923 and pUR-KP1025, respectively. The general modules of both transposons were In35::ISCR1::blaCTX-M-2-Tn1000-like-Tn2*-ISKpn11-12-13 variable module-ΔTn21. In Tn7057 there was ΔIS10R-catA2 associated with an additional ISKpn13. Both plasmids belonged to IncC type 2 and ST3. pUR-KP0923 was 167 138 bp in length and had a 37 926-bp resistance island at position 4 (RI-4). Plasmid pUR-KP1025 was 168 128 bp with a RI-4 of 36 222 bp. CONCLUSION: This report describes the molecular nature of two transposons (Tn7057 and Tn7058) harbouring blaCTX-M-2 that reside in IncC type 2 ST3 plasmids. These transposons mediate resistance to oxyimino-cephalosporins, gentamicin and, in the case of Tn7057, chloramphenicol. CTX-M-2 is an important extended-spectrum ß-lactamase (ESBL) to South American epidemiology. It is remarkable that despite being only two plasmid sequences, the information revealed here could contribute to a better understanding of the resistance islands from IncC type 2 plasmids.

Diversity of the epidemiology of carbapenemase-producing Enterobacteriaceae in long-term acute care rehabilitation settings from an area of hyperendemicity, and evaluation of an intervention bundle.

BACKGROUND: Long-term acute care rehabilitation facilities (LTACRFs) are affected by carbapenem-resistant Enterobacteriaceae (CRE) in endemic areas. However, the contribution of different subpopulations of patients has not been investigated in these settings. AIM: To study the epidemiology of CRE in an LTACRF, and the effect of an infection control intervention. METHODS: A surveillance programme was implemented in a large Italian LTACRF. The intervention included screening for CRE carriage at admission and weekly (for negative patients), and enforcement of contact precautions plus cohorting (in wards and rehabilitation areas) for presumed and confirmed carriers. Prevalence and incidence of CRE colonization and the number of CRE bacteraemias were monitored over one year. FINDINGS: Overall, 1084 patients underwent screening (adherence 89.8%). At admission, 11.6% of patients were colonized, and 9.9% of those negative at admission subsequently became colonized. These percentages were significantly higher among patients with severe brain injuries (SBIs) who were exposed to a higher intensity of care (44.1% vs 8.6% and 63.5% vs 6.8%, respectively). The majority of CRE bacteraemias occurred in the SBI ward. The intervention was associated with a decline in the incidence of CRE colonization in the SBI ward (from 17.7 to 7.2 acquisitions/100 at-risk patient-weeks), but not in other wards. All CRE isolates were Klebsiella pneumoniae carbapenemase-producing K. pneumoniae. CONCLUSIONS: A peculiar CRE epidemiology was observed in a LTACRF from Italy, with very high rates of carriage and cross-transmission in SBI patients. A simplified infection control bundle was effective at reducing the incidence of CRE colonization in the SBI ward.

A simple phenotypic method for screening of MCR-1-mediated colistin resistance.

OBJECTIVES: To evaluate a novel method, the colistin-MAC test, for phenotypic screening of acquired colistin resistance mediated by transferable mcr-1 resistance determinants, based on colistin MIC reduction in the presence of dipicolinic acid (DPA). METHODS: The colistin-MAC test consists in a broth microdilution method, in which colistin MIC is tested in the absence or presence of DPA (900 µg/mL). Overall, 74 colistin-resistant strains of Enterobacteriaceae (65 Escherichia coli and nine other species), including 61 strains carrying mcr-1-like genes and 13 strains negative for mcr genes, were evaluated with the colistin-MAC test. The presence of mcr-1-like and mcr-2-like genes was assessed by real-time PCR and end-point PCR. For 20 strains, whole-genome sequencing data were also available. RESULTS: A ≥8-fold reduction of colistin MIC in the presence of DPA was observed with 59 mcr-1-positive strains, including 53 E. coli of clinical origin, three E. coli transconjugants carrying MCR-1-encoding plasmids, one Enterobacter cloacae complex and two Citrobacter spp. Colistin MICs were unchanged, increased or at most reduced by twofold with the 13 mcr-negative colistin-resistant strains (nine E. coli and four Klebsiella pneumoniae), but also with two mcr-1-like-positive K. pneumoniae strains. CONCLUSIONS: The colistin-MAC test could be a simple phenotypic test for presumptive identification of mcr-1-positive strains among isolates of colistin-resistant E. coli, based on a ≥8-fold reduction of colistin MIC in the presence of DPA. Evaluation of the test with a larger number of strains, species and mcr-type resistance determinants would be of interest.

Draft Genome Sequence of Clostridium difficile Belonging to Ribotype 018 and Sequence Type 17.

Clostridium difficile, belonging to ribotype 018 (RT018), is one of the most prevalent genotypes circulating in hospital settings in Italy. Here, we report the draft genome of C. difficile CD8-15 belonging to RT018, isolated from a patient with fatal C. difficile-associated infection.

Severe obesity prevalence in 8- to 9-year-old Italian children: a large population-based study.

BACKGROUND AND OBJECTIVES: Little information is available on severe obesity in childhood. This study estimates the prevalence of severe obesity in 8- to 9-year-old children resident in Italy and its association with gender, age, geographical area and parents' nutritional status and education using the World Health Organization (WHO) and International Obesity Task Force (IOTF) criteria. SUBJECTS/METHODS: A nationally representative sample of grade 3 Italian students was measured in 2010 (N=42,431) using standardized instruments and methodology. Severe obesity in children was assessed using definitions provided by the WHO and by the IOTF. Prevalence was estimated within categories of sociodemographic variables and their independent effects were estimated using multivariate logistic regression. RESULTS: The estimated prevalence of severe obesity in 2010 was 4.5% (95% confidence interval (CI): 4.2-4.7) according to the WHO definition and 2.7% (95% CI: 2.5-2.9) with IOTF cutoffs. These values were slightly lower than those observed in 2008. The prevalence was higher in males, in 8-year-old children and in the South. Parental low education and high body mass index were strongly associated with childhood severe obesity. CONCLUSION: According to the definition used, between 30,000 and 50,000 children aged 8-9 years suffer severe obesity in Italy.

Pseudomonas aeruginosa infection in cystic fibrosis caused by an epidemic metallo-ß-lactamase-producing clone with a heterogeneous carbapenem resistance phenotype.

An epidemic IMP-13 metallo-ß-lactamase (MBL)-producing Pseudomonas aeruginosa clone, causing infections and even large outbreaks in Italian critical care settings, was detected in a young cystic fibrosis patient. In this patient, the chronic infection was sustained by distinct clonal sub-populations of the MBL-producing P. aeruginosa clone, either susceptible or resistant to carbapenems. These findings underscore the importance of infection prevention practices in cystic fibrosis settings and pose an important diagnostic and therapeutic challenge.

BMI from nutritional surveillance of 8-9 years old children in Tuscany (Italy).

INTRODUCTION: The latest increase in childhood obesity focused attention on the important consequences that this phenomenon may have on public health in relationship to the increasing risk that an obese child may become an obese adult. To deal with this problem, there is necessary to assess systematically the distribution of childhood nutritional status at different levels: international, regional and local. In this paper are presented data on underweight, overweight and obesity prevalence in third grade primary school children, aged 8/9 years in Tuscany (2008) and its distribution in relationship to the demographic breadth of their place of residence. METHODS: Data from statistic sample of 2109 (1.091 males, 1.018 females), 8/9 years school-children were collected; weight and height were measured using standardised personnel and instruments. Exact month age was calculated between the data of measurement and that of birth. Body Mass Index (BMI) classes were calculated using Cole et al.'s epidemiologic cut-off for children and adolescents. Residence areas were divided into four classes based on the number of inhabitants (< 10.000; 10.000-50.000; > 50.000; > 50.000 metropolitan). RESULTS: The prevalence of underweight was 0.88% (0.76% in males and 1.01% in females), the prevalence of overweight was 23.43% (22.33% in males and 24.65% in females), the prevalence of obese was 7.95% (9.08% in males, 6.70% in females). The lowest prevalence of obese (6.46%) was found in towns with over 50.000 residents (metropolitan). CONCLUSION: The obesity prevalence in Tuscany children is still lower than that of the Italian National Survey, while the overweight prevalence it's the same. Obesity prevalence (10.71%) is higher in municipalities with low residents number (< 10.000).

Underweight and overweight among children and adolescents in Tuscany (Italy). Prevalence and short-term trends.

INTRODUCTION: The recent increase in both childhood obesity and adolescent anorexia nervosa in developed countries has underlined the important consequences that these trends may have on public health, as there is an increased risk that these conditions may become chronic diseases in adulthood. Therefore, it is necessary to monitor prevalence rates and trends in thinness and overweight (including obesity) among children and adolescents at different levels: international, national and sub-national. Since 2001/2002, a nutritional surveillance system has been implemented in the Tuscany Region to estimate the nutritional status and lifestyles of children and adolescents. The main objectives were to assess the prevalence of thinness, overweight and obesity among Tuscan children and adolescents and to provide baseline information on the prevalence of thinness, for the first time calculated according to the new international definitions, for geographical comparisons and descriptions of time trends. METHODS: Independent cross-sectional sample surveys were conducted in 2002, 2004 and 2006 in Tuscany, North-Central Italy. Data were collected from stratified two-stage cluster samples of children aged 9 years (n = 3,048 in 2002 and n = 1,430 in 2006) and of adolescents aged 11-13-15 years (n = 1,066, n = 1185 and n = 1,160 in 2004 and n = 1,189, n = 1,211 and n = 1,178 in 2006, respectively). Weights and heights of primary school children were measured by means of standardized methods, while those of adolescents were self-reported. Decimal age was calculated from the date of birth to the date of measurement. Body Mass Index classes were calculated according to the International Obesity Task Force standards. Instead of the term underweight in children, we used the term thinness, which the World Health Organization uses to mean low Body Mass Index for age in adults and adolescents. According to Cole's recently published cut-offs for thinness, we divided our Body Mass Index values below 18.5 into three grades. RESULTS: This study presents data on the prevalence of different grades of nutritional status (thinness, normal weight, overweight and obesity) among Tuscan school-aged children from primary to high school (9-11-13-15-y-old), assessed by means of Body Mass Index, according to international definitions. From 2002 to 2006 the prevalence of thinness among children aged 9 years decreased from 4.6% to 4.2%, and the prevalence of normal weight from 63.7% to 62.4%; the prevalence of overweight (including obesity) rose from 31.7% to 33.4%. From 2004 to 2006, among pre-adolescents aged 11 years, the prevalence of thinness declined from 11.0% to 10.1%; the prevalence of normal weight rose from 68.4% to 70.2%, and the prevalence of overweight declined from 20.7% to 19.6%. Among adolescents aged 13 years, the prevalence of thinness declined from 9.8% to 8.0%; the prevalence of normal weight rose from 73.5% to 74.0%, and the prevalence of overweight from 16.8% to 17.9%. Among adolescents aged 15 years, the prevalence of thinness declined from 9.8% to 8.7%, and the prevalence of normal weight from 77.0% to 71.6%, while the prevalence of overweight rose from 13.3% to 19.7%. The 2006 data showed that the trend in the prevalence of overweight (including obesity) tended to decrease with age for both sexes, though more markedly in girls (from 34.0% at 9-y of age to 12.2% at 15-y of age) than in boys (from 32.8% at 9-y of age to 22.8% at 13-y of age to 27.5% at 15-y of age). By contrast, the prevalence of thinness increased with age in girls (from 4.9% at 9-y of age to 14,1% at 15-y of age), while boys presented a similar low prevalence at 9 and 15-y of age (3.3% and 3.1%), doubling the values at 11 and 13-y of age (7.5% and 6.5%). The trend in the prevalence of normal weight increased with age from 62.4% at 9-y of age to 74.0% at 13-y of age and to 71.6% at 15-y of age. Boys displayed a higher prevalence than girls only at 9-y of age (63.9% vs. 61.0%). DISCUSSION: The results of this study allow us to analyze data from the nutritional surveillance system in Tuscany using recent definitions of Body Mass Index cut-off points among children, pre-adolescents and adolescents. As a rule, the trend in the prevalence of overweight (including obesity) among girls from 9-y to 15-y-old strongly decreased, while the prevalence of thinness increased. In boys, this decrease was less marked and the prevalence of thinness displayed an irregular trend, with an increment from 9-y to 11-y-old and a decrease from 13-y to 15-y old. The trend in the prevalence of normal weight increased with age, with a higher prevalence among boys than girls.

A pilot trial of low-dose naltrexone in primary progressive multiple sclerosis.

A sixth month phase II multicenter-pilot trial with a low dose of the opiate antagonist Naltrexone (LDN) has been carried out in 40 patients with primary progressive multiple sclerosis (PPMS). The primary end points were safety and tolerability. Secondary outcomes were efficacy on spasticity, pain, fatigue, depression, and quality of life. Clinical and biochemical evaluations were serially performed. Protein concentration of beta-endorphins (BE) and mRNA levels and allelic variants of the mu-opiod receptor gene (OPRM1) were analyzed. Five dropouts and two major adverse events occurred. The remaining adverse events did not interfere with daily living. Neurological disability progressed in only one patient. A significant reduction of spasticity was measured at the end of the trial. BE concentration increased during the trial, but no association was found between OPRM1 variants and improvement of spasticity. Our data clearly indicate that LDN is safe and well tolerated in patients with PPMS.

Disease-modifying drugs in childhood-juvenile multiple sclerosis: results of an Italian co-operative study.

OBJECTIVE: Immunomodulatory drugs (IDs) (interferon beta (IFNgamma) and glatiramer acetate (GA)) reduce relapse rate and disease progression in relapsing remitting multiple sclerosis (RRMS) but extensive data are not available on the effectiveness and tolerability of these drugs in childhood or adolescence. The aim of this study was to evaluate the impact of IFNbeta and GA in MS patients treated before 16 years of age. METHODS: A research group (Immunomodulatory Treatment of Early onset MS (ITEMS)) was promoted in Italy to collect a large series of patients affected by clinically definite and RRMS and treated with IDs before 16 years of age. Fifteen centres recognized subjects suitable for inclusion: 76 patients (52 females) were collected with a mean age at onset of 12.4 (SD 2.5) years, a mean disease duration of 18.6 (SD 14.7) and a relapse rate of 3.1 (SD 2.9). RESULTS: Results were evaluated in 65 (45 females) subjects with a pretreatment and a treatment duration >3 months: 38 were treated with IFNbeta-1a once weekly (Avonex), 18 with IFNbeta three times weekly (16 with Rebif, 2 with Betaferon) and nine with GA (Copaxone). The mean pretreatment period was respectively 20, 18 and 9.2 months. The treatment duration lasted respectively 23.3, 40.7 and 33.3 months. The mean annualized relapse rate decreased dramatically during the treatment: from 2.4 to 0.4 in the Avonex group, from 3.2 to 0.8 in the Rebif-Betaferon group and from 2.8 to 0.25 in the GA group. The mean final EDSS scores were respectively (in brackets the initial scores): 1.3 (1.4), 1.6 (1.8) and 0.6 (1.1). In the whole group, the final score was unchanged or reduced in all subjects except eight. Clinical side effects were recorded in 41/65 subjects (mainly in subjects treated with IFNbeta), abnormal laboratory findings were observed in 13/65 subjects: they were transient in most cases. IFNgamma was stopped in six cases: in four because of inefficacy and in two cases because of side effects. CONCLUSIONS: Sixty-five clinically definite MS subjects were treated during childhood or adolescence with IDs. The treatment reduced the relapse rate and the progression of the disease in most cases. Side effects were common in subjects treated with IFNbeta but were well tolerated in most cases.