Respiratory Isolates Research Articles

Comparative expression of Toll-like receptors and inflammatory cytokines in pigs infected with different virulent porcine reproductive and respiratory syndrome virus isolates

BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) is largely responsible for heavy economic losses in the swine industry worldwide because of its high mutation rate and subsequent emergence of virulent strains. However, the immunological and pathological responses of pigs to PRRSV strains with different virulence have not been completely elucidated.MethodsTwenty-four piglets were divided into 4 groups (n = 6 each) and inoculated with highly pathogenic PRRSV isolate BB0907 (HP), low pathogenic PRRSV NT0801 (LP), LP derivative strain NT0801-F70 (LP-der), and DMEM medium (control), respectively. The changes in TLR2, 3, 7, and 8 gene expression and TNF-α, IL-1β, IL-6, IFN-γ, and IL-10 secretion were evaluated using real-time PCR and ELISA at 6, 9, and 15 days post inoculation (d.p.i.). The cytokine levels were evaluated in the supernatants of porcine alveolar macrophages (PAMs) and peripheral blood mononuclear cells (PBMCs) following stimulation with LTA, poly(I:C), CL097, and PRRSV individually.ResultsHP caused more severe clinical signs and pathological lesions in swine than LP and LP-der had almost no virulence compared with LP. The serum levels of IL-1β, IL-6, TNF-α, and IFN-γ were increased in HP-infected piglets, which were greater than in those infected with LP or LP-der. The mRNA levels of TLR3, 7, and 8 were significantly up-regulated in PAMs in HP-infected pigs compared to those in groups LP and LP-der. Furthermore, TNF-α and IL-1β secretion in PAMs from group LP was statistically greater than those from the control group after stimulation with either poly(I:C) or CL097. Meanwhile, TNF-α, IL-1β, and IL-6 levels in CL097-stimulated PBMCs from HP-infected pigs were markedly higher than those from the LP- and LP-der-infected groups.ConclusionsWe found that HP was a stronger inducer of TLR 3, 7, and 8 expression and IL-1β, IL-6, TNF-α, and IFN-γ production compared to LP and LP-der. HP enhanced production of TNF-α, IL-1β, and IL-6 in PBMCs following CL097-stimulation more than LP and LP-der, whereas LP enhanced the secretion of TNF-α and IL-1β in poly(I:C)- and CL097-stimulated PAMs. Our data regarding cellular reactivity to different isolates should be useful in the development of more efficacious vaccines.

Absence of Mycobacterium intracellulare and Presence of Mycobacterium chimaera in Household Water and Biofilm Samples of Patients in the United States with Mycobacterium avium Complex Respiratory Disease

Recent studies have shown that respiratory isolates from pulmonary disease patients and household water/biofilm isolates of Mycobacterium avium could be matched by DNA fingerprinting. To determine if this is true for Mycobacterium intracellulare, household water sources for 36 patients with Mycobacterium avium complex (MAC) lung disease were evaluated. MAC household water isolates from three published studies that included 37 additional MAC respiratory disease patients were also evaluated. Species identification was done initially using nonsequencing methods with confirmation by internal transcribed spacer (ITS) and/or partial 16S rRNA gene sequencing. M. intracellulare was identified by nonsequencing methods in 54 respiratory cultures and 41 household water/biofilm samples. By ITS sequencing, 49 (90.7%) respiratory isolates were M. intracellulare and 4 (7.4%) were Mycobacterium chimaera. In contrast, 30 (73%) household water samples were M. chimaera, 8 (20%) were other MAC X species (i.e., isolates positive with a MAC probe but negative with species-specific M. avium and M. intracellulare probes), and 3 (7%) were M. avium; none were M. intracellulare. In comparison, M. avium was recovered from 141 water/biofilm samples. These results indicate that M. intracellulare lung disease in the United States is acquired from environmental sources other than household water. Nonsequencing methods for identification of nontuberculous mycobacteria (including those of the MAC) might fail to distinguish closely related species (such as M. intracellulare and M. chimaera). This is the first report of M. chimaera recovery from household water. The study underscores the importance of taxonomy and distinguishing the many species and subspecies of the MAC.

GRP-086 Identifying New Tuberculosis Cases Through Pharmacy Dispensing Records in Prof Dr Fernando Fonseca Hospital, Portugal

BackgroundControlling and preventing tuberculosis (TB) continues to be a major public healthcare challenge. Pharmacy and clinical records can thus contribute with important information concerning newly-diagnosed inpatients, treatment regimens and resistant...

Activities of carbapenem and comparator agents against contemporary US Pseudomonas aeruginosa isolates from the CAPITAL surveillance program

Antimicrobial susceptibilities of contemporary Pseudomonas aeruginosa clinical isolates were determined from the CAPITAL 2010 surveillance program. Isolates were collected from 100 sites throughout the USA and Puerto Rico, and included isolates representing a range of patient demographics and infection types. A total of 2722 isolates were tested for susceptibility to a broad spectrum of agents, with susceptibilities ranging from 98.8% for colistin to 74% for levofloxacin. Doripenem was the most active carbapenem agent, with 88.6% of isolates susceptible, in comparison with 78.1% and 84.6% for imipenem and meropenem, respectively. Lower respiratory tract isolates and isolates from the intensive care unit setting were the least susceptible overall. Resistance rates were typically highest in lower respiratory tract isolates, with the exception of urinary tract isolates, which displayed the highest resistance for levofloxacin. Overall, multidrug-resistant isolates comprised 14.8% of the total sample population.

Detection of clonally related Escherichia coli isolates producing different CMY β-lactamases from a cystic fibrosis patient

This study reports details on Escherichia coli isolates recovered from a cystic fibrosis (CF) patient in order to understand how this pathogen adapts to and resists broad-spectrum antipseudomonal therapy in this context. Five E. coli isolates were obtained from various clinical samples (airways, urine or dialysis catheter) over a 7 month period covering a double-lung transplantation. All isolates were analysed in terms of clonality [enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus sequence typing], virulence profiles (phylogroup and search for 15 virulence genes), growth patterns (morphotype, biofilm-forming ability and growth rate), hypermutability and antimicrobial susceptibility, with molecular characterization of β-lactamases and porins. The five isolates shared similar ERIC-PCR profiles and sequence types (ST1193) and exhibited the same virulence profile. The respiratory isolates were strong mutators, exhibited mucoid or small-colony morphotypes, exhibited strong biofilm-forming ability and grew slowly compared with the others. All isolates were highly resistant to ceftazidime. The respiratory isolates showed reduced susceptibility to cefepime and high resistance to aztreonam. The patient had received a 31 day course of ceftazidime/aztreonam until transplantation. All isolates harboured the same wild-type chromosomal AmpC. A CMY-2 enzyme was detected in the non-respiratory isolates. The respiratory isolates harboured L293S and V211A/L293S new CMY-2 variants, which were designated CMY-94 and CMY-95, respectively. OmpF porin loss was observed in the non-respiratory isolates. Our study shows that, similarly to Pseudomonas aeruginosa, E. coli can undergo phenotypic and genomic changes in the CF context. For the first time, we identified an in vivo expanded-spectrum evolution of the CMY-2 β-lactamase, during bacterial persistence in the CF lung.

Temporal changes in respiratory adenovirus serotypes circulating in the greater Toronto area, Ontario, during December 2008 to April 2010

BackgroundCertain adenovirus serotypes cause severe infections, especially in children. It is important to monitor temporal changes in serotypes causing clinical disease. The objective of this study was to document circulating respiratory adenovirus serotypes by sequencing adenovirus culture isolates from the Greater Toronto Area, Ontario, during December 2008 to April 2010.MethodsNucleic acid extraction was performed on 90 respiratory tract adenovirus culture isolates. PCR amplification was conducted with primers targeting the adenovirus hexon gene hypervariable region 7. Sanger sequencing and phylogenetic analyses were performed to determine serotype identities.ResultsAmong 90 clinical respiratory isolates sequenced, eight different serotypes were identified. Serotype 3 (34, 38%), serotype 2 (30, 30%), and serotype 1 (14, 16%) isolates were most common; serotypes 5, 6, 11, 17 and 21 were also observed. Seventeen (50%) of the 34 HAdV-3 isolates were identified between December 2008 and February 2009, while none were identified from December 2009 to February 2010. Between December 2008 and April 2009, the two most common serotypes were HAdV-3 and HAdV-2, detected in 18 (53%) and 8 (24%) of the 34 cultures isolates, respectively. However, from December 2009 to April 2010, there was an increase in HAdV-2, which became the most prevalent serotype, detected in 10 (50%) of the 20 isolates identified (p = 0.05).ConclusionsThere was a gradual shift in prevailing adenovirus serotypes during the 17 month study period, from predominantly HAdV-3 to HAdV-2. If an adenovirus vaccine were to be broadly implemented, multiple serotypes should be included.

<i>Bordetella holmesii</i>: Comparison of Two Isolates from Blood and a Respiratory Sample

Interest in Bordetella holmesii is increasing, but very little is known about this bacterium, which can be isolated from both blood and respiratory samples. In this study, we compared a B. holmesii isolate from the blood sample of an adult with bacteremia with another isolate from a nasopharyngeal swab from an adult with whooping cough syndrome. Genetic analysis was carried out, targeting relevant genes, and virulence properties were studied in cellular and animal models. Our genomic analysis provided no evidence of traits specific to either blood or respiratory isolates of B. holmesii. Neither isolate was cytotoxic to human tracheal epithelial cells. Both isolates were only weakly invasive and they did not persist within epithelial cells for less than 48 h.

An update on the MLST scheme for Pasteurella multocida

Pasteurella multocida is a cause of economically important diseases in almost all domestic livestock species, as well as wildlife. While a range of typing methods have traditionally been used, the development of a Multi-Locus Sequence Typing (MLST) scheme in 2010 represented the first standardised, sequence based, Web supported typing scheme. The initial scheme (termed the RIRDC MLST scheme) was based on 63 avian isolates from diseased Australian poultry and three international reference strains, which formed 29 Sequence Types (STs). The MLST database (http://pubmlst.org/pmultocida_rirdc/) now contains data from over 560 isolates that form 220 STs. The use of the scheme in published studies to date has demonstrated some key points: A) the highly clonal nature of haemorrhagic septicaemia (HS) isolates; B) bovine respiratory isolates are typically very distinct from HS isolates; C) evidence of host/niche association (i.e. some STs are associated with specific hosts); and D) the distinct genotype of P. multocida isolates of capsule type B from calf pleuritis and peritonitis cases in New Zealand. The continued use of this MLST scheme by research groups around the world will add to our understanding of the population structure and host associations of this major veterinary pathogen.

Bovine coronaviruses from the respiratory tract: Antigenic and genetic diversity

BoCV isolated from respiratory tract, nasal swab and broncho alveolar washing fluid samples were evaluated for genetic and antigenic differences. These BoCV from the respiratory tract of healthy and clinically ill cattle with BRD signs were compared to reference and vaccine strains based on Spike protein coding sequences and VNT using convalescent antisera. Based on this study, the BoCV isolates belong to one of two genomic clades (clade 1 and 2) which can be differentiated antigenically. The respiratory isolates from Oklahoma in this study were further divided by genetic differences into three subclades, 2a, 2b, and 2c. Reference enteric BoCV strains and a vaccine strain were in clade 1. Currently available vaccines designed to control enteric disease are based on viruses from one clade while viruses isolated from respiratory tracts, in this study, belong to the other clade.

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Introduction: Anecdotally, an increase in early isolates of Serratia, Pseudomonas, Acinetobacter, Citrobacter and Enterobacter (SPACE) species from respiratory specimens was noted in patients admitted to our trauma-orthopedics-vascular (TOV) intensive care unit (ICU). Given the guideline-recommended empiric anti-infective therapy for early pneumonia may not provide sufficient coverage for SPACE organisms, specifically Pseudomonas and AmpC-producing Enterobacteriaceae, patients may receive inadequate initial anti-infective therapy. Hypothesis: We hypothesize that SPACE respiratory isolates are being identified early, defined < 5 days following admission, in our TOV ICU patients. Methods: Medical records of 212 patients with gram negative respiratory isolates (GNRI) and admitted to TOV ICU from November 1st, 2009 to May 31st, 2012 were retrospectively reviewed. The primary aim of the study is to evaluate the number of GNRI with SPACE species. The secondary aim is to determine the times between admission, intubation, and respiratory cultures with SPACE organisms. Results: Cultures from 83% of patients grew at least one SPACE organism. In approximately 1/3 of patients with GNRI, a SPACE organism was isolated < 5 days following admission. The median times between first GNRI positive for a SPACE organism and hospital and ICU admissions were 7.3 days and 6.6 days, respectively. The median time between first intubation and GNRI positive for SPACE organism was 6.4 days. Conclusions: Of TOV ICU patients with GNRI, the majority were SPACE organisms. Although the median time from admission to SPACE respiratory isolate was > 5 days, about 1/3 of patients with GNRI yielded a SPACE organism < 5 days, which remains concerning given current anti-infective recommendations for early pneumonia. Patient specific characteristics of these patients with early pulmonary SPACE isolates will be collated for trends or risk factors. These data will facilitate both the use of targeted anti-infective therapy in our ICU setting as well as the development of quality improvement initiatives that may optimize anti-infective utilization.

Antimicrobial Resistance Genotype Trend and Its Association with Host Clinical Characteristics in Respiratory Isolates of Haemophilus influenzae

Background: β-Lactam resistance genotype trends in clinical isolates of Haemophilus influenzae and their correlation with the clinical background were analyzed. Methods: Five hundred and ten respiratory isolates of H. influenzae collected during the period 2002–2009 were classified by PCR into gBLNAS (genotype for β-lactamase-negative ampicillin-susceptible), gBLNAR (genotype for β-lactamase-negative ampicillin-resistant) and 3 other genotypes. The associations with host clinical data and antimicrobial susceptibility were analyzed in all 144 isolates between 2008 and 2009. Results: The 8-year trend analysis detected an increase in gBLNAR with a decrease in gBLNAS. The probability of being a causative pathogen did not differ between genotypes. Host clinical characteristics such as age and gender did not differ with gBLNAR or gBLNAS, but the underlying respiratory diseases did differ. gBLNAR was found at the highest rate in 83% of isolates from patients with nontuberculous mycobacteriosis. In contrast, gBLNAR accounted for as little as 33% of isolates from chronic obstructive pulmonary disease. Conclusions: There were no differences in the pathogenicity of gBLNAR and gBLNAS. The underlying respiratory diseases may be related to the resistance genotype.

Mycobacterial Interspersed Repetitive-Unit–Variable-Number Tandem-Repeat (MIRU-VNTR) Genotyping of Mycobacterium intracellulare for Strain Comparison with Establishment of a PCR-Based Database

Strain comparison is important to population genetics and to evaluate relapses in patients with Mycobacterium avium complex (MAC) lung disease, but the "gold standard" of pulsed-field gel electrophoresis (PFGE) is time-consuming and complex. We used variable-number tandem repeats (VNTR) for fingerprinting of respiratory isolates of M. intracellulare from patients with underlying bronchiectasis, to establish a nonsequence-based database for population analysis. Different genotypes identified by PFGE underwent species identification using a 16S rRNA gene multiplex PCR. Genotypes of M. intracellulare were confirmed by internal transcribed spacer 1 (ITS1) sequencing and characterized using seven VNTR primers. The pattern of VNTR amplicon sizes and repeat number defined each specific VNTR type. Forty-two VNTR types were identified among 84 genotypes. PFGE revealed most isolates with the same VNTR type to be clonal or exhibit similar grouping of bands. Repetitive sequence-based PCR (rep-PCR) showed minimal pattern diversity between VNTR types compared to PFGE. Fingerprinting of relapse isolates from 31 treated patients using VNTR combined with 16S multiplex PCR unambiguously and reliably distinguished different genotypes from the same patient, with results comparable to those of PFGE. VNTR for strain comparison is easier and faster than PFGE, is as accurate as PFGE, and does not require sequencing. Starting with a collection of 167 M. intracellulare isolates, VNTR distinguished M. intracellulare into 42 clonal groups. Comparison of isolates from different geographic areas, habitats, and clinical settings is now possible.

Epidemiology and clinical significance of non-tuberculous mycobacteria isolated from pulmonary specimens

A tertiary university medical centre in northern Israel. To evaluate the clinical significance of non-tuberculous mycobacteria (NTM) isolated from pulmonary specimens. Clinical and microbiological data were collected from patient files. Cases were classified as definite, probable and possible NTM. Between 2004 and 2010, 215 cases with respiratory isolates of NTM were identified. Mycobacterium xenopi was the most common species (n = 84, 39.1%), followed by M. simiae (n = 52, 24.2%). A total of 170 (79.1%) cases were classified as possible and 24 (11.2%) as probable NTM. Only 21 (9.8%) cases were considered definite NTM, the majority of which were M. kansasii and M. avium complex. M. xenopi and M. simiae are the most prevalent species of NTM isolated from respiratory samples in northern Israel. However, most of these isolates represent colonisation. Of the relatively small number of clinically significant isolates, M. kansasii and M. avium complex were the most common.

Comparison of phenotypic and virulence genes characteristics in human and chicken isolates of Proteus mirabilis

The objective of this work is to compare the phenotypic and virulence genes characteristics in human and chicken isolates of Proteus mirabilis. The bacterial examination of 50 livers of individual broilers, marketed by four major outlets, revealed a high recovery of P. mirabilis (66%), and a low recovery frequency of Salmonella spp. (4%), Serratia odorifera (2%), Citrobacter brakii (2%), and Providencia stuartii (2%). The phenotypic biochemical characterization of the recovered 33 chicken isolates of P. mirabilis were compared to 30 human isolates (23 urinary and six respiratory isolates). The comparison revealed significant differences in the presence of gelatinase enzyme (100% presence in chicken isolates versus 91·3 and 83·3% presence in human urinary and respiratory isolates, respectively, P<0·05). The H2S production occurred in 100% of chicken isolates versus 95·6 and 66·7% presence in human urinary and respiratory isolates, respectively, P<0·05). The other 17 biochemical characteristics did not differ significantly among the three groups of isolates (P>0·05). Two virulence genes, the mrpA and FliL, were having a typical 100% presence in randomly selected isolates of P. mirabilis recovered from chicken livers (N = 10) versus isolates recovered from urinary (N = 5) and respiratory specimens of humans (N = 5) (P>0·05). The average percentage similarity of mrpA gene nucleotide sequence of poultry isolates to human urinary and respiratory isolates was 93·2 and 97·5-%, respectively. The high similarity in phenotypic characteristics, associated with typical frequency of presence of two virulence genes, and high similarity in sequences of mrpA gene among poultry versus human P. mirabilis isolates justifies future investigations targeting the evaluation of adaptable pathogenicity of avian Proteus mirabilis isolates to mammalian hosts.

Bovine coronaviruses from the respiratory tract

Bovine coronaviruses (BoCV) have been associated with bovine respiratory diseases (BRD) in cattle pulled for treatment in feedlots and from healthy cattle in the US with BoCV identified by virus isolation from nasal swab and bronchoalveolar lavage (BAL) samples, and also by seroconversions for the detection of active infections. Bovine coronaviruses have been identified in pneumonic lungs from field cases, often in combination with other viruses and bacteria including Mycoplasma spp. Experimentally, BoCV infections have caused respiratory tract lesions. It has been suggested there is a dual tropism of BoCV for the respiratory and enteric tracts. Historically, BoCV was associated with neonatal enteric disease and adult winter dysentery. Respiratory isolates were identified later and focused on post-weaning cattle entering feedlots. Control measures are limited. There are USDA-licensed killed and modified live virus (MLV) vaccines for BoCV; however, these are for control of the enteric disease in cattle. There are no USDA-licensed vaccines for control of BoCV associated BRD in cattle. The purpose of the study was to compare BoCVisolates from the respiratory tract of cattle in Oklahoma to reference respiratory tract and enteric strains via antigenic and genetic procedures.

Serotypes, antimicrobial susceptibility, and beta-lactam resistance mechanisms of clinical Haemophilus influenzae isolates from Bulgaria in a pre-vaccination period

To determine the serotypes, antimicrobial susceptibility, and beta-lactam resistance mechanisms of Haemophilus influenzae strains isolated from invasive and respiratory tract infections (RTIs) prior to the introduction of Haemophilus influenzae type b (Hib) vaccination in Bulgaria. A total of 259 isolates were serotyped by polymerase chain reaction. Susceptibility to antibiotics and beta-lactamase production were determined, and DNA sequencing of the ftsI gene was performed for ampicillin non-susceptible strains. The invasive H. influenzae infections in children were mainly due to serotype b (94.5% in meningitis and 88.9% in other invasive cases). Non-typeable strains (97.4%) were the most frequently found H. influenzae strains in RTIs both in children and adults. Non-susceptibility to ampicillin occurred in 22% of all strains. Ceftriaxone and levofloxacin were the most active agents tested. Ampicillin resistance occurred in 34.4% of invasive strains, and beta-lactamase production was the only mechanism found. Among respiratory tract isolates, ampicillin non-susceptible strains (18%) were classified into the following groups: beta-lactamase-positive, ampicillin-resistant (BLPAR) strains (7.2%); beta-lactamase-negative, ampicillin-non-susceptible (BLNAR) strains (8.2%); and beta- lactamase-positive, amoxicillin-clavulanate-resistant (BLPACR) strains (2.6%). Among 21 BLNAR and BLPACR strains there were 9 different patterns of multiple-amino acid substitutions in penicillin-binding protein 3. Of these, most isolates (81.0%) belonged to group II, defined by the Asn526Lys substitution. Beta-lactamase production was more common among invasive strains than in respiratory isolates. BLNAR and BLPACR H. influenzae were found only among respiratory tract isolates.

AWARE Ceftaroline Surveillance Program (2008–2010): Trends in Resistance Patterns Among Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis in the United States

Ceftaroline fosamil, the prodrug form of the active metabolite ceftaroline, is a new broad-spectrum parenteral cephalosporin with antibacterial activity against the prevalent respiratory pathogens Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and Staphylococcus aureus. Bacterial resistance surveillance (5330 isolates) was conducted in the United States between 2008 and 2010 to assess the in vitro activity of ceftaroline and comparator antibacterial agents against invasive respiratory isolates of S. pneumoniae (3329 isolates), H. influenzae (1545 isolates), and M. catarrhalis (456 isolates). All organisms were cultured from patient infections in 71 US hospital laboratories and were submitted to a central reference monitor for broth microdilution testing by Clinical and Laboratory Standards Institute reference methods. Against S. pneumoniae, ceftaroline inhibited 98.7% of strains at the susceptible breakpoint of ≤ 0.25 µg/mL (50% minimum inhibitory concentration [MIC(50)], 0.01 µg/mL; 90% MIC [MIC(90)], 0.12 µg/mL) and was 16-fold more active than ceftriaxone (MIC(90), 2 µg/mL). Among 70 ceftriaxone-resistant pneumococcal isolates, all were inhibited by ≤ 0.5 µg/mL of ceftaroline. Haemophilus influenzae (MIC(50), ≤ 0.008 µg/mL; MIC(90), 0.015 µg/mL) and M. catarrhalis (MIC(50), 0.06 µg/mL; MIC(90), 0.12 µg/mL) were very susceptible to ceftaroline regardless of β-lactamase production. Whereas the high-level of activity of ceftaroline was maintained against S. pneumoniae and H. influenzae from 2008 through 2010, increased rates of nonsusceptibility were observed for amoxicillin/clavulanate, erythromycin, and levofloxacin among S. pneumoniae and for trimethoprim/sulfamethoxazole and azithromycin among H. influenzae. In summary, ceftaroline resistance surveillance (Assessing Worldwide Antimicrobial Resistance Evaluation [AWARE] Program) in the United States (2008-2010) documented in vitro sustained potency and spectrum against Gram-positive and Gram-negative pathogens known to cause community-acquired bacterial pneumonia.

An investigation into the prevalence and outcome of patients admitted to a pediatric intensive care unit with viral respiratory tract infections in Cape Town, South Africa

To describe the prevalence and outcome of patients admitted to a pediatric intensive care unit with viral respiratory tract infections. Retrospective descriptive study. Pediatric intensive care unit in a tertiary pediatric hospital situated in Cape Town, South Africa. All children (n = 195; 20% pediatric intensive care unit admissions) with positive respiratory viral isolates between April 1 and December 31, 2009. None. Demographic, clinical, laboratory, and outcome data were recorded from medical folders. Complete data were available for 175 patients (median age [interquartile range] 4.7 months [2.3-12.9 months]; 49% male). One hundred four (59.4%) patients had comorbid conditions; 30 (17%) were HIV-infected. Rhinovirus (n = 76 [39%]), respiratory syncytial virus (n = 54 [27.7%]), adenovirus (n = 30 [15.4%]), influenza A (n = 26 [13.3%]), parainfluenza (n = 23 [11.8%]), and human metapneumovirus (n = 12 [6.2%]) were most commonly isolated. Ninety-five infections (51.4%) were isolated >48 hrs after admission. Seasonal patterns were identified for respiratory syncytial virus, human metapneumovirus, and influenza A, whereas others occurred throughout the year. Twenty-five patients (14.3%) had more than one viral isolate. Presumed bacterial coinfection, which occurred in 68 (39%) patients (18 [26.5%] HIV-infected), was associated with significantly longer pediatric intensive care unit and hospital stays but not with mortality. Twenty patients died (11%, standardized mortality ratio 0.64). High Pediatric Index of Mortality scores, HIV exposure and infection, nosocomial infection, and influenza A infection were associated with mortality. Viral respiratory tract infection is common in this pediatric intensive care unit associated with significant morbidity and mortality, which may relate to the high burden of comorbidity and HIV.

Pulmonary Disease Caused by Rapidly Growing Mycobacteria: A Retrospective Study of 44 Cases in Japan

Background: The features of pulmonary disease caused by rapidly growing mycobacteria (RGM) have not been sufficiently documented. Objectives: To establish these features, we retrospectively evaluated 44 patients. Methods: We screened respiratory isolates at the National Toneyama Hospital (Osaka, Japan) between 2003 and 2007. Diagnosis was based on the latest guidelines of the American Thoracic Society. The patients were classified into 3 types according to their radiographic findings: fibrocavitary, nodular bronchiectatic and unclassified variant. Results: We obtained 1,348 nontuberculous mycobacteria respiratory isolates from 1,187 patients, including 119 RGM isolates from 100 patients. Forty-four of these 100 patients were definitively diagnosed with respiratory disease due to RGM. The most common pathogen was Mycobacteriumabscessus, which accounted for 65.9% of cases, followed by Mycobacterium fortuitum at 20.5%. There was a statistically significant difference in smoking history between patients infected with these 4 RGM species (excluding those with an unknown smoking history; p = 0.039). The overall evaluation of radiographic findings revealed 18.2% as fibrocavitary, 43.2% as nodular bronchiectatic and 38.6% as unclassified variants in these 44 patients. There was a significant difference in radiographic findings between the 4 RGM species (p = 0.002). There was also a significant difference in radiographic findings between M. abscessus and M. fortuitum infected patients (p = 0.022). Conclusions: Patients with M. abscessus seem to have less of a smoking history and more frequent nodular bronchiectatic radiographic patterns than patients with M. fortuitum. In contrast, fibrocavitary patterns might be more frequent with M. fortuitum infection.

Differentiation in Quinolone Resistance by Virulence Genotype in Pseudomonas aeruginosa

Pseudomonas aeruginosa is a leading pathogen that has become increasingly resistant to the fluoroquinolone antibiotics due to widespread prescribing. Adverse outcomes have been shown for patients infected with fluoroquinolone-resistant strains. The type III secretion system (TTSS) is a major virulence determinant during acute infections through the injection of effector toxins into host cells. Most strains exhibit a unique TTSS virulence genotype defined by the presence of either exoS or exoU gene encoding two of the effector toxins, ExoS and ExoU, respectively. Specific TTSS effector genotype has been shown previously to differentially impact virulence in pneumonia. In this study, we examined the relationship between TTSS effector genotype and fluoroquinolone resistance mechanisms in a collection of 270 respiratory isolates. We found that a higher proportion of exoU+ strains were fluoroquinolone-resistant compared to exoS+ strains (63% vs 49%, p = 0.03) despite its lower overall prevalence (38% exoU+ vs 56% exoS+). Results from sequencing the quinolone resistance determining regions (QRDRs) of the 4 target genes (gyrA, gyrB, parC, parE) indicated that strains containing the exoU gene were more likely to acquire ≥2 mutations than exoS+ strains at MICs ≤8 µg/ml (13% vs none) and twice as likely to have mutations in both gyrA and parC than exoS+ strains (48% vs 24% p = 0.0439). Our findings indicate that P. aeruginosa strains differentially develop resistance-conferring mutations that correlate with TTSS effector genotype and the more virulent exoU+ subpopulation. Differences in mutational processes by virulence genotype that were observed suggest co-evolution of resistance and virulence traits favoring a more virulent genotype in the quinolone-rich clinical environment.