The global crisis of multidrug resistance: how to face healthcare associated infections without effective antibiotics?

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Association of total serum cholesterol with functional outcome following home care rehabilitation in Italian patients with stroke.

BACKGROUND: Stroke is a disabling disease. In elderly populations, stroke is the third leading cause of death and the primary cause of reduction in or loss of functional ability and personal autonomy. Possible associations between levels of total serum cholesterol (TC) and both incidence of stroke and functional outcomes after rehabilitation are still under study. OBJECTIVE: To detect positive and negative prognostic factors associated with functional outcomes in first-time stroke patients admitted to an integrated home care rehabilitative program. METHODS: This study enrolled 141 patients with a first-time stroke who were admitted to a home care rehabilitation program. Primary outcome measures were the Barthel activities of daily living (ADL) and mobility indices at the beginning and end of the rehabilitative treatment. The impact of TC and other demographic and clinical variables was analyzed using bivariate and multivariate logistic regression analyses. RESULTS: Age and Short Portable Mental Status Questionnaire (SPMSQ) score were negatively associated with functional outcome. In contrast, elevated TC was positively associated with a better home rehabilitative treatment outcome. Barthel index score at admission was negatively associated with outcomes assessed by the Barthel ADL index and age with outcomes assessed by the Barthel mobility index. In a multivariate logistic regression analysis, SPMSQ score and elevated TC were significantly associated with outcome. Specifically, higher SPMSQ scores were negatively associated with better rehabilitative treatment outcomes, whereas elevated TC was positively associated. CONCLUSIONS: Elevated TC seems to be associated with better functional outcomes in patients with first-time stroke

Simultaneous detection of Escherichia coli, Salmonella enterica, Listeria monocytegenes and Bacillus cereus by oligonucleotide microarray

Background: Traditional laboratory methods to detect pathogenic bacteria are time consuming and laborious. Therefore, it is essential to use powerful and reliable molecular methods for quick and simultaneous detection of microbial pathogens. Objectives: The current study aimed to evaluate the capability and efficiency of 23S rDNA sequence for rapid and simultaneous detection of four important food-borne pathogens by an oligonucleotide microarray technique. Materials and Methods: The 23S rDNA sequences of Escherichia coli, Salmonella enterica, Listeria monocytogenes and Bacillus cereus were obtained from GenBank databases and used to design the oligonucleotide probes and primers by Vector NTI software. Oligonucleotide probes were placed on a nylon membrane and hybridization was performed between probes and 23S rDNA digoxigenin-labeled polymerase chain reaction (PCR) products. Hybridization signals were visualized by NBT/BCIP color development. Results: Positive hybridization color was produced for Escherichia coli, Salmonella enterica, Listeria monocytogenes and Bacillus cereus. The oligonucleotide microarray detected all bacterial strains in a single reaction in less than five hours. The sensitivity of the performed microarray assay was 103 cfu/mL for each species of pathogen. No cross reaction was found between the tested bacterial species. Conclusions: The obtained results indicated that amplification of 23S rDNA gene followed by oligonucleotide microarray hybridization is a rapid and reliable method to identify and discriminate foodborne pathogens tested under the study

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

Characterization of endemic Shigella boydii strains isolated in Iran by serotyping, antimicrobial resistance, plasmid profile, ribotyping and pulsed-field gel electrophoresis

Background: Shigellosis is one of the major causes of morbidity in children with diarrhea in Iran. The present study was undertaken to characterize apparently sporadic Shigella boydii strains isolated from pediatric patients in Iran. Findings: Ten S. boydii strains isolated from pediatric cases of gastroenteritis and acute diarrhea in Tehran between December 2002 and November 2003 were submitted to serotyping, antimicrobial susceptibility testing, plasmid profile analysis, ribotyping and pulsed field gel electrophoresis (PFGE). Seven isolates were attributed to serotype 2, whereas the remaining three belonged to serotypes 14, 18, 19, respectively. Six drug resistance phenotypes (R1 to R6) were defined with R4 - streptomycin (STR), ampicillin (AMP), sulfamethoxazole-trimethoprim (SXT) - being the most prevalent. Plasmid analysis resulted in seven different plasmid profiles with one to five DNA bands. All strains, but one, shared the same ribotype, but PFGE differentiated them in four groups. Conclusion: Based upon ribotyping and PFGE results, endemic circulation of S. boydii in Tehran, Iran, could be attributed to a few clones. Resistance pattern and plasmid profile analysis proved to be very effective in discriminating apparently unrelated strains of S. boydi

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

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

Antibacterial activity of Borago officinalis and Brassica juncea aqueous extracts evaluated in vitro and in situ using different food model systems

The present study was undertaken to characterize the antibacterial activity of the aqueous extracts (AEs) obtained from the leaves of Borago officinalis L. and Brassica juncea L. The antagonistic activity was evaluated against several bacteria (42 strains of Listeria monocytogenes, 35 strains of Staphylococcus aureus, 38 strains of Enterobacter spp. and 18 strains of Salmonella enterica) commonly associated with foodborne diseases by paper disc diffusion method. The susceptibility to the plant extracts was strain specific. Thirty-five strains (7 L. monocytogenes, 11 S. aureus, 1 S. Enteritidis, 1 S. Veneziana, 7 Enterobacter hormaechei, 5 Enterobacter cloacae, 1 Enterobacter sakazakii and 2 Enterobacter amnigenus) were sensitive to both AEs. The activity of B. juncea AE towards the Gram-positive strains was generally higher than that observed for B. officinalis (45 and 22 strains inhibited by B. juncea and B. officinalis, respectively), while an opposite trend was registered against the Gram-negative strains (22 and 35 strains inhibited by B. juncea and B. officinalis, respectively). The highest inhibition was displayed by B. juncea AE against E. sakazakii 23A. B. officinalis AE showed the same minimum inhibitory concentration (MIC) (10 mg/mL) for the majority of the most sensitive strains, while the MIC of B. juncea AE was different for each bacterial species and the lowest concentration was registered to inhibit enterobacteria (3.1 mg/mL). After 1-year storage in different thermal conditions (room temperature, 4 C and 20 C), both AEs lost their inhibitory power. The extracts did not show cellular toxicity when tested against sheep erythrocytes. Hence, B. officinalis and B. juncea AEs were effective as natural antibacterial substances. AEs were tested in situ in three food model systems (meat, fish and vegetable) at two concentrations, but only when added at a concentration 10-fold higher than that showing definite efficacy in vitro (100 and 31 mg/mL for B. officinalis and B. juncea, respectively), they inhibited the growth of the sensitive strains, even though the cells were still viable after 24 h. The influence of AEs on the volatile organic compounds (VOCs) composition of the food models was analysed by gas chromatography/mass spectrometry. The different levels of alcohols, aldehydes, esters, hydrocarbons, ketones and phenol registered, showed a consistent effect of B. officinalis and B. juncea AEs on the VOCs of the food models. However, the sniffing assay found only B. juncea AE impacting consistently the final aroma of the food models

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