KPC-3-producing bacteria are endemic in many countries but only recently became apparent their wide distribution in different Portuguese hospitals. The aim of this study is to characterize genetic backgrounds associated with blaKPC−3 among Klebsiella pneumoniae isolates recently identified on non-hospitalized patients in Portugal. Twenty KPC-producing K. pneumoniae identified between October 2014 and November 2015 in three different community laboratories were characterized. Isolates were mainly from patients from long-term care facilities (n = 11) or nursing homes (n = 6), most of them (75%) previously hospitalized in different Portuguese hospitals. Standard methods were used for bacterial identification and antibiotic susceptibility testing. Carbapenemase production was assessed by the Blue-Carba test, and identification of bla genes was performed by PCR and sequencing. Epidemiological features of KPC-producing K. pneumoniae included population structure (XbaI-PFGE, MLST and wzi sequencing), genetic context (mapping of Tn4401), and plasmid (replicon typing, S1-PFGE, and hybridization) analysis. All K. pneumoniae isolates produced KPC-3, with two MDR K. pneumoniae epidemic clones representing 75% of the isolates, namely ST147 (wzi64/K14.64, February–November 2015) and ST15 (two lineages exhibiting capsular types wzi19/K19 or wzi93/K60, July-November 2015). Other sporadic clones were detected: ST231 (n = 3; wzi104), ST348 (n = 1; wzi94) and ST109 (n = 1, wzi22/K22.37). blaKPC−3 was identified within Tn4401d in all isolates, located in most cases (80%) on cointegrated plasmids (repAFIA+repAFII+oriColE1;105-250 kb) or in 50 kb IncN plasmids. In conclusion, this study highlights a polyclonal structure of KPC-3-producing K. pneumoniae and the predominance of the ST147 clone among non-hospitalized patients in Portugal, linked to platforms still unnoticed in Europe (blaKPC−3-Tn4401d-IncFIA) or firstly reported (blaKPC−3-Tn4401d-IncN). This scenario underlines the recent penetration of successful mobile genetic elements in previously circulating MDR K. pneumoniae lineages (mainly ST147 and ST15) in Portugal, rather than the importation of the global lineages from clonal group 258.

The presence of bats in caves, attics, ceilings, and roofs is important epidemiologically as they can increase the chance of human acquisition of pathogens, including Histoplasma capsulatum. Brazilian urban areas contain many species of bats, especially insectivorous bats, that are attracted by a wide range of readily available food and shelter. From August 2003 to December 2008, we analysed 2427 bats in the São Paulo State region. Homogenates of the livers and spleens of the bats were plated on specific medium to identify animals infected with H. capsulatum. The fungus was isolated from 87 bats (3·6%). The infected bats were identified as Molossus molossus (74), Nyctinomops macrotis (10), Tadarida brasiliensis (1), Molossus rufus (1) and Eumops glaucinus (1), all insectivorous species. The data presented are a relevant contribution to the epidemiology of H. capsulatum in densely populated urban areas such as in São Paulo State, especially since histoplasmosis is not included in the mandatory disease notification system.

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New arr alleles emerged in class 1 integrons from a clinical Pseudomonas aeruginosa strain (arr-4) and four Klebsiella pneumoniae strains (arr-5) in Brazil/American continent. arr-4 was preceded by aacA7-catB3, whereas arr-5 was the unique cassette. The putative proteins shared 75% (Arr-5) and 78% (Arr-4) identities with Arr-2.Rifampin resistance is a notable global health problem concern (11), since it is the front line drug for treating tuberculosis (4) and preventing meningococcal diseases (13). More than 90% of rifampin resistance is due to mutational alterations in an 81-bp rifampin resistance determining region of the rpoB gene (9). However, resistance may also arise by horizontal acquisition of arr genes, which code for ADP-ribosyltransferases responsible for the drug inactivation. The arr-1 gene was first described in the Mycobacterium smegmatis chromosome (10). Subsequently, arr-2 and arr-3 alleles were described as gene cassettes in class 1 integrons present in gram-negative isolates from Europe and Asia (1,6,8,14), showing high-level rifampin MICs ranging from 32 to Ͼ256 g/ml.We report here the identification and characterization of two new arr alleles, arr-4 and arr-5, and the emergence of this class of gene in the American continent as gene cassettes from class 1 integrons present in clinical Pseudomonas aeruginosa and Klebsiella pneumoniae strains.One P. aeruginosa (PS1111) and four K. pneumoniae (K56, K224, K688, and K830) isolates recovered from different inpatients of a Brazilian hospital (Rio de Janeiro city) in 2005 tested positive for class 1 integron signatures (3). DNA macrorestriction using SpeI, followed by pulsed-field gel electrophoresis, was performed as previously described (3) and demonstrated four distinct profiles among the K. pneumoniae strains. Sequence analysis of the class 1 integron variable regions showed the presence of a 453-bp open reading frame, encoding a predicted protein of 150 amino acid residues in all integrons. These sequences from the K. pneumoniae strains and from isolate PS1111 shared 78% amino acid identity, and 75 and 78% amino acid identity, respectively, with ADP-ribosyltransferase Arr-2 (Fig. 1). Therefore, these open reading frames were named arr-4 (PS1111) and arr-5 (the K. pneumoniae strains). The genes were assigned to be parts of cassettes due to the presence of attC sites. The arr-4 gene cassette was preceded by a aacA7-catB3 array, which confer resistance to aminoglycosides and chloranphenicol, respectively, and arr-5 was the unique cassette inserted in all K. pneumoniae integrons. The arr-4 gene cassette is 516 bp long and presented an attC site only 22 bp in length due to the loss of 2L and 2R simple sites (Fig. 2). The arr-5 gene cassette is 555 bp, presenting a classical attC of 61 bp, encompassing the core site sequences that make up the left-hand and right-hand consensus sites composed of 1L, 2L, 1R, and 2R simple sites (Fig. 2) (12).arr-2 and arr-3 gene cassettes share 99% nucleotide identity and, moreover, present identical attC site sequence...

Hospital associated methicillin-resist Staphylococcus aureus has long been associated to outbreaks in the hospital environment. In this work, we investigated an outbreak of Hospital associated methicillin-resist Staphylococcus aureus carrying the Panton-Valentine leukocidin gene, which occurred in a large community hospital in Rio de Janeiro, Brazil.