Agent Of Pneumonia Research Articles

Mitochondrial Dysfunction in Aged Macrophages and Lung during Primary Streptococcus pneumoniae Infection is Improved with Pirfenidone

Pneumococcal infections remain a leading cause of death in older adults, with the most serious cases occurring in persons ≥65 years of age. There is an urgent need to investigate molecular pathways underlying these impairments and devise new therapeutics to modulate innate immunity. The goal of our current study is to understand the impact of chronological aging on mitochondrial function in response to Streptococcus pneumoniae, a causative agent of bacterial pneumonia. Using chronologically aged murine models, our findings demonstrate that decreased ATP production is associated with dysregulated mitochondrial complex expression, enhanced oxidative stress, diminished antioxidant responses, and decreased numbers of healthy mitochondria in aged adult macrophages and lung in response to S. pneumoniae. Pre-treatment of aged macrophages with pirfenidone, an anti-fibrotic drug with antioxidant and anti-inflammatory properties, improved mitochondrial function and decreased cellular oxidative stress responses. In vivo administration of pirfenidone decreased superoxide formation, increased healthy mitochondria number, improved ATP production, and decreased inflammatory cell recruitment and pulmonary oedema in aged mouse lung during infection. Taken together, our data shed light on the susceptibility of older persons to S. pneumoniae and provide a possible therapeutic to improve mitochondrial responses in this population.

Epidemiological and clinical analysis of community-acquired Mycoplasma pneumonia in children from a Spanish population, 2010-2015

ResumenIntroducción y objetivosMycoplasma pneumoniae (MP) es uno de los agentes etiológicos más comunes de las neumonías adquiridas en la comunidad (NAC) en niños. Objetivo: describir las características clínicas y epidemiológicas, tratamiento y evolución de los pacientes con NAC por MP (NACM) en un hospital terciario de Valencia, España.Material y métodosSe revisaron retrospectivamente las historias clínicas de los niños < 14 años con NACM entre enero de 2010 y diciembre de 2015. Los pacientes con evidencia radiológica de neumonía y confirmación microbiológica de MP (PCR de exudado nasofaríngeo y/o anticuerpos IgM específicos frente a MP) se consideraron NACM.ResultadosUn total de 162 pacientes se diagnosticaron de NACM; mediana de edad de 6 años (rango intercuartílico: 4-9 años). La proporción de pruebas positivas para MP en pacientes con NAC, así como el uso empírico de macrólidos, aumentó progresivamente con la edad. Hubo un pico de casos en 2011 y en 2015, con un máximo de casos en julio, agosto, noviembre y diciembre. El patrón radiológico más frecuente fue el infiltrado segmentario (62,3%), mientras que 22 (13,6%) presentaron derrame pleural. Los niños con NACM desarrollaron una clínica leve, con poca elevación de parámetros inflamatorios. Se inició tratamiento empírico con un macrólido en el 68,5% de los casos. La necesidad de ingreso hospitalario fue inversamente proporcional a la edad del paciente.ConclusionesSegún este estudio, los niños con NAC de mayor edad tuvieron la mayor proporción de infección por MP, siendo poco sintomáticos y con escasa elevación de parámetros inflamatorios, pudiéndose beneficiar del tratamiento empírico con macrólidos. Por consiguiente, conocer la epidemiología de un área geográfica podría ser importante para el abordaje de las NAC en niños.

Btk inhibitor ibrutinib reduces inflammatory myeloid cell responses in the lung during murine pneumococcal pneumonia

BackgroundStreptococcus pneumoniae is a major causative agent in community-acquired pneumonia and sepsis. Overwhelming lung inflammation during pneumococcal pneumonia may hamper lung function. Ibrutinib is an irreversible inhibitor of Bruton’s tyrosine kinase (Btk), a key signaling protein controlling the activation of various immune cells, including macrophages and neutrophils. The aim of this study was to determine whether ibrutinib treatment ameliorates acute lung inflammation during pneumococcal pneumonia.MethodsMice were treated orally with ibrutinib and the effect on acute pulmonary inflammation elicited by the gram-positive bacterial cell wall component lipoteichoic acid (LTA) and during ceftriaxone-treated pneumococcal pneumonia was assessed.ResultsTreatment with ibrutinib prior to and after intranasal LTA instillation reduced alveolar macrophage activation, neutrophil influx, cytokine release and plasma leakage into the lung. Postponed treatment with ibrutinib supplementing antibiotic therapy during ongoing pneumococcal pneumonia did not impair bacterial killing in lung, blood and spleen. In this setting, ibrutinib reduced alveolar macrophage and systemic neutrophil activation and substantially diminished further monocyte and neutrophil influx in the lung. In vitro, ibrutinib inhibited macrophage TNF secretion and neutrophil activation upon LTA and pneumococcal stimulation.ConclusionsTaken together, these data indicate that the Btk inhibitor ibrutinib reduces inflammatory myeloid cell responses during acute pulmonary inflammation evoked by LTA and antibiotic-treated pneumococcal pneumonia and suggest that ibrutinib has the potential to inhibit ongoing lung inflammation in an acute infectious setting.

Етіологія ензоотичної пневмонії свиней та стратегії боротьби з нею в фермерських господарствах Запорізької та Полтавської областей

Porcine enzootic pneumonia is a multifactorial disease, it leads to large and significant economic losses in pig farms and it remains a relevant veterinary problem. The involvement of new etiological agents in the association of pathogens and the constantly changing conditions on the farms lead to the fact that previously developed control schemes and methods of disease lost their effectiveness. Therefore, the identification of all the infectious agents that make up the association, as well as the concomitant factors that led to the occurrence of the disease in each case is the only correct way to control measures. When we studied the situation in pig farms in the south and in the central region of Ukraine, we found that mycoplasmas were the main etiological agent for Porcine enzootic pneumonia. We noted that mycoplasmosis as a monoinfection was recorded only in 9-12 % of cases. In most outbreaks Mycoplasmas were only members of associations along with Porcine reproductive and respiratory syndrome infection (6%), Type 2 Circovirus infection (6%), enterbacterias (19%) and pasterellas (59%). The course of the disease was also aggravated by the stress of early weaning of piglets and violations of the conditions of feeding and animal welfare. The most difficult enzootic pneumonia occurred in those farms where mycoplasmosis and pasteurellosis were exacerbated by the presence of pathogens of actinobacillus pleuropneumonia. In such farms the average daily weight gain decreased in piglets by a third and feed consumption increased to 28 %. Piglets on affected farms were ill with obvious respiratory clinical signs. And 82% of dead or killed piglets had signs of fibrinous pleuropneumonia or catarrhal pneumonia. In those farms where we recorded associated respiratory infections, we also found reproductive-neonatal infections of pigs. In such farms treatment and prophylactic measures using traditional antimicrobial special remedies and monovaccines, did not give the expected effect. We proposed the use of autovaccines for associated enzootic pneumonia.

Prevalence of vaginal Group-B-Streptococcus, antibiotic and antigen sensitivity amongst parturients at the federal medical center Owerri, Nigeria

Background: Group B streptococcus (GBS) is one of the principal agents of early onset neonatal sepsis, pneumonia and meningitis with significant morbidity for newborns and parturients. Aim: This study was done to determine the prevalence of vaginal group B streptococcus amongst parturients in FMC Owerri. It also aimed to elucidate the antibiotic sensitivity of the strains of GBS isolated while comparing the sensitivity of antigen detection tests of GBS to culture. Patients and Methods: This cross-sectional study was done at the labour ward and lying-in units of the Federal Medical Center, Owerri,from February 2015 to October, 2015. It involved one hundred and eighty (180) women recruited systematically. Two swab samples, high vaginal and rectal were collected from the parturients intrapartum and cultured. The babies were examined and weighed immediately after delivery, seen on the second and tenth days after delivery for features of fever or any other complaints. Results: The overall prevalence of GBS amongst parturients was found to be 6.1%. Prevalence of vaginal colonization was 3.3% and that of the rectum was 2.8%, (2c=0.1282, df=1,p=0.502). Neonates with colonized mothers all had complaints of fever postpartum which was statistically significant (=55.86, df=1, p<0.001). All the microbes showed 100% sensitivity to penicillin G, erythromycin, gentamycin and ceftriaxone. Antibiotic resistance was discovered in augmentin (67%), Cloxacillin (55.6%) and ofloxacin (33.3%). Antigen detection tests gave Sensitivity=100%, specificity=96.4%, positive predictive value (PPV) = 64.7%, negative predictive value (NPV) =100%. Detection in both vaginal (=60.290,p=0.000) and rectal (=50.799,p=0.000) samples were statistically significant. Conclusion: The prevalence of GBS in parturients in the Centre is low. Strains of GBS isolated amongst the sampled population in FMC Owerri had a high sensitivity to penicillin G, erythromycin, gentamycin and 3 rd generation cephalosporins. Antigen detection method for screening of GBS though very sensitive is however not specific with a significant false positive detection rate due to antigen cross reaction. It is however recommended that antigen detection be employed as intrapartum measure in high risk cases to reduce turnover time whilst supported by culture results later. There was associated significant neonatal sequaelae and further research to establish causal relationship will be advised. Keywords: Prevalence, Group B streptococcus, colonization, antibiotic, antigen sensitivity

Community acquired pneumonia of viral-bacterial etiology in young people: actual aspects of clinical and laboratory diagnostics

Community acquired pneumonia is one of the most topical acute respiratory diseases, which is caused by high incidence rate, especially in organized groups of people, constantly changing microbial flora, the increasing resistance of bacteria to antibacterial drugs, difficulties in etiological diagnostics, and the possibility of life-threatening complications and fatalities. An innovative algorithm of etiological diagnostics of community acquired pneumonia is suggested, which includes immunochromatography express-tests of sputum for viruses detection, bacteriological sputum culture, immunoenzyme assay of blood and polymerase chain reaction of sputum and blood serum. It is shown that standard bacteriological sputum culture did not allow us to reveal causative agents of community-acquired pneumonia timely and precisely in most cases, whereas the application of more comprehensive etiological diagnostics enabled us to reveal the causative agent in the majority of the examined patients. And express methods provided an opportunity to verify the infectious agent within 1-2 days and administer early effective etiotropic treatment. It is demonstrated that modern community acquired viral bacterial pneumonia has a number of clinical and laboratory features depending on the revealed viral agents. These features may be used as additional diagnostic criteria of the disease, especially when modern methods of etiological diagnostics are unavailable. The obtained results showed the effectiveness of the use of immunochromatography express-tests of sputum, polymerase chain reaction of sputum and blood serum, and immunoenzyme assay of blood. As a result of statistical analysis, a number of characteristic clinical and laboratory predictors of certain viral-bacterial associations of modern community acquired pneumonia was determined.

Transmission Electron Microscopy Improves the Diagnostic Sensitivity in Nonbacterial Etiology of Severe Pneumonia: A Retrospective Study

BackgroundSevere pneumonia is responsible for great mortality and morbidity worldwide, and early-applied effective anti-infective therapy can improve the prognosis of patients. However, identification of infectious agents in severe pneumonia remains a major challenge so far. In this study, the potential utility of transmission electron microscopy (TEM) in detecting nonbacterial pathogens in patients with severe pneumonia was retrospectively evaluated.Materials and MethodsA total of 106 patients diagnosed with severe pneumonia at our hospital from September 2015 to December 2017 were included, and their baseline clinical characteristics were collected. Nonbacterial infectious agents detected by TEM in bronchoalveolar lavage fluid (BALF) and serological tests were summarized. The detection rates were further compared between TEM and serological tests.ResultsBALF examination under the transmission electron microscope revealed 24 viruses, 16 mycoplasmas, 18 chlamydia, 2 fungi and 74 bacteria in 99 samples, among which 61 samples were mixed infections. The combined use of serological tests and TEM significantly improved the detection rate of nonbacterial infectious agents in patients with severe pneumonia.ConclusionsOur data support that implementation of TEM could improve the sensitivity for detecting viruses, atypical pathogens and mixed infections in BALF from patient of severe pneumonia. Therefore, TEM may be used as an auxiliary diagnostic method of other microbiological tests in severe pneumonia.

1880. Does the Label Matter: Initial Use of Newly Approved Antimicrobial Agents in Community Hospitals Without Robust Antimicrobial Stewardship Programs

BackgroundAntimicrobial agents are often used for indications not approved by the Food and Drug Administration (FDA). While this can be appropriate at times, they may be used in settings with no published experience. Judicious use of new agents is critical for conserving their utility. This study characterized initial use after FDA approval of select antimicrobial agents in community hospitals without robust antimicrobial stewardship programs (ASP).MethodsInitial use of systemic antimicrobials approved by the FDA since 2014 was retrospectively reviewed at 6 community hospitals (50–350 beds). Up to 10 charts of first use were reviewed per drug at each hospital. Time from FDA approval to first administration was measured for the following agents: ceftazidime–avibactam, ceftolozane–tazobactam, dalbavancin, isavuconazonium, oritavancin, and peramivir. Clinical indications, prescribing service, and empiric uses were recorded.ResultsMean time from FDA approval to first administration ranged from 12 (tedizolid) to 26 months (dalbavancin). Of frequently used agents (Figure 1), adherence to initial FDA indications ranged from 7% (ceftolozane–tazobactam for complicated urinary tract infection) to 100% (tedizolid for acute bacterial skin and soft-tissue infection). Pneumonia (35%) and osteomyelitis (35%) were the most common off-label indications for ceftazidime–avibactam. Pneumonia was the most common off-label indication for ceftolozane–tazobactam (78%). The most common off-label indications for oritavancin were osteomyelitis (14%) and bacteremia (11%). Infectious Diseases was the main prescribing service for all agents (range 74–95%). Use of Gram-positive agents was mostly empiric whereas Gram-negative agents were targeted against specific pathogens. ConclusionNewly approved antimicrobial agents were used at these six community hospitals within 1–2 years after FDA approval. Agents with primarily Gram-positive activity were more often used for FDA approved indications. Given frequent use of novel Gram-negative agents for pneumonia, there is need for early trials to determine their role for this indication. In the meantime, ASP’s should consider off-label indications such as pneumonia when developing local criteria for use.Disclosures All authors: No reported disclosures.

General overview of the detection of Mycoplasma hyopneumoniae DNA by quantitative polymerase chain reaction in diagnostic cases submitted to the Iowa State University Veterinary Diagnostic Laboratory from 2004 to 2016

Mycoplasma hyopneumoniae (Mhp) is the etiologic agent of enzootic pneumonia and a major causative agent of the porcine respiratory disease complex. This study summarizes and describes the general diagnostic trends on Mhp detection by quantitative polymerase chain reaction (qPCR) in cases submitted to the Iowa State University Veterinary Diagnostic Laboratory from 2004 to 2016. The following variables were included in the analysis: animal age, geographic location, sample type, season, and submission year. The overall frequency of detection found was 27.04% and ranged from 17.9% to 40.7%. Lung homogenate and bronchial swabs had a greater Mhp qPCR detection rate than other sample types (P &lt; .001) followed by bronchoalveolar lavage (P &lt; .001), while oral fluids had the lowest Mhp detection rate (P &lt; .001). The fall season had a greater percentage of positive Mhp qPCR results when compared to other seasons (P &lt; .001), while spring had the lowest percentage. Finishing-age pigs had a greater percentage of Mhp qPCR detection when compared to other age groups (P &lt; .001), while suckling pigs had the lowest percentage (P &lt; .001).

Utilization of macrophage extracellular trap nucleotides by Mycoplasma hyopneumoniae.

Mycoplasma hyopneumoniae is the causative agent of enzootic pneumonia in swine, an important disease worldwide. It has finite biosynthetic capabilities, including a deficit in de novo nucleotide synthesis. The source(s) for nucleotides in vivo are unknown, but mycoplasmas are known to carry membrane-bound nucleases thought to participate in the acquisition of nucleotides from host genomic DNA. Recent research has demonstrated that neutrophils can produce extracellular traps (NETs), chromatin NETs decorated with granular proteins to interact with and eliminate pathogens. We hypothesized that M. hyopneumoniae could utilize its membrane nuclease to obtain nucleotides from extracellular traps to construct its own DNA. Using the human monocytic cell line THP-1, we induced macrophage extracellular traps (METs), which are structurally similar to NETs. The thymidine analogue ethynyl deoxyuridine (EdU) was incorporated into THP-1 DNA and METs were induced. When incubated with M. hyopneumoniae, METs were degraded and the modified nucleotide label could be co-localized within M. hyopneumoniae DNA. When the nucleases were inhibited, MET degradation and nucleotide transfer were also inhibited. Controls confirmed that the EdU originated directly from the METs and not from free nucleotides arising from intracellular pools released during extrusion of the chromosomal DNA. M. hyopneumoniae incorporated labelled nucleotides more efficiently when 'fed' on METs than from free nucleotides in the medium, suggesting a tight linkage between nuclease degradation of DNA and nucleotide transport. These results strongly suggest that M. hyopneumoniae could degrade extracellular traps formed in vivo during infection and incorporate those host nucleotides into its own DNA.

Age-related changes in the levels and kinetics of pulmonary cytokine and chemokine responses to Streptococcus pneumoniae in mouse pneumonia models

Streptococcuspneumoniae is a major human pathogen at the extremes of age. The elderly are particularly vulnerable to S.pneumoniae, the most common causative agent of bacterial pneumonia in this population. Despite the availability of vaccines and antibiotics, mortality rates associated with pneumococcal pneumonia in this age group remain high. In light of globally increasing life-expectancy, a better understanding of the patho-mechanisms of elderly pneumococcal pneumonia, including alterations in innate immune responses, is needed to develop improved therapies.In this study we aimed at investigating how increased susceptibility to pneumococcal infection relates to inflammation kinetics in the aged mouse pneumonia model by determining pulmonary cytokine and chemokine levels and comparing these parameters to those measured in young adult mice.Firstly, we detected overall higher pulmonary cytokine and chemokine levels in aged mice. However, upon induction of pneumococcal pneumonia in aged mice, delayed production of certain analytes, such as IFN-γ, MIG (CXCL9), IP-10 (CXCL10), MCP-1 (CCL2), TARC (CCL17) and MDC (CCL22) became apparent. In addition, aged mice were unable to control excess inflammatory responses: while young mice showed peak inflammatory responses at 20 h and subsequent resolution by 48 h post intranasal challenge, in aged mice increasing cytokine and chemokine levels were measured. These findings highlight the importance of considering multiple time points when delineating inflammatory responses to S.pneumoniae in an age-related context. Finally, correlation between pulmonary bacterial burden and cytokine or chemokine levels in young mice suggested that appropriately controlled inflammatory responses support the host to fight pneumococcal infection.

Surface proteins mhp390 (P68) contributes to cilium adherence and mediates inflammation and apoptosis in Mycoplasma hyopneumoniae

Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia (EP) and responsible for major economic losses in global swine industry. After colonization of the respiratory epithelium, M. hyopneumoniae elicits a general mucociliary clearance loss, prolonged inflammatory response, host immunosuppression and secondary infections. Until now, the pathogenesis of M. hyopneumoniae is not completely elucidated. This present study explores the pathogenicity of mhp390 (P68, a membrane-associated lipoprotein) by elucidating its multiple functions. Microtitrer plate adherence assay demonstrated that mhp390 is a new cilia adhesin that plays an important role in binding to swine tracheal cilia. Notably, mhp390 could induce significant apoptosis of lymphocytes and monocytes from peripheral blood mononuclear cells (PBMCs), as well as primary alveolar macrophages (PAMs), which might weaken the host immune response. In addition, mhp390 contributes to the production of proinflammatory cytokines, at least partially, via the release of IL-1β and TNF-α. To the best of our knowledge, this is the first report of the multiple functions of M. hyopneumoniae mhp390, which may supplement known virulence genes and further develop our understanding of the pathogenicity of M. hyopneumoniae.

Pulmonary fungal granulomas and fibrinous pneumonia caused by different hypocrealean fungi in reptiles

In contrast to fungal dermatitis, fungal glossitis and disseminated visceral mycosis, fungal infection of the lung has so far rarely been described in reptiles. Pulmonary fungal granulomas were diagnosed histopathologically within the scope of post mortem examinations. Fragments of the 18S-internal transcribed spacer1-5.8S rDNA (SSU-ITS1-5.8S) and 28S rDNA (LSU), including domains (D)1 and D2 as well as the protein coding gene translation elongation factor 1 alpha (TEF) were used for phylogenetical analysis after isolation of the fungal pathogen by culturing. Ten reptiles, including lizards (n = 6), snakes (n = 1), crocodilians (n = 2) and tortoises (n = 1) presented with pulmonary fungal granulomas (n = 8) and fibrinous pneumonia (n = 2) caused by different non-clavicipitaceous and clavicipitaceous species of the order Hypocreales. Purpureocillium lavendulum (n = 2) and Metarhizium robertsii (n = 1) as the etiologic agents of pneumonia in reptile species are described for the first time. Fungal pulmonary granulomas caused by clavicipitaceous fungi (n = 6) were all associated with disseminated visceral mycosis as well as oral fungal granulomas (n = 4) and/or fungal dermatitis (n = 1). Differing infection routes being likely for clavicipitaceous and non-clavicipitaceous fungal pathogens. A potential zoonotic health risk should be taken into account during necropsy or lung sampling in live reptiles with pulmonary fungal granulomas, since human infections, mainly keratitis and sclerokeratitis, caused by Beauveria bassiana, Metarhizium robertsii and Purpureocillium lilacinum, have occasionally been described.

Pathogenic variability among Pasteurella multocida type A isolates from Brazilian pig farms

BackgroundPasteurella multocida type A (PmA) is considered a secondary agent of pneumonia in pigs. The role of PmA as a primary pathogen was investigated by challenging pigs with eight field strains isolated from pneumonia and serositis in six Brazilian states. Eight groups of eight pigs each were intranasally inoculated with different strains of PmA (1.5 mL/nostril of 10e7 CFU/mL). The control group (n = 12) received sterile PBS. The pigs were euthanized by electrocution and necropsied by 5 dpi. Macroscopic lesions were recorded, and swabs and fragments of thoracic and abdominal organs were analyzed by bacteriological and pathological assays. The PmA strains were analyzed for four virulence genes (toxA: toxin; pfhA: adhesion; tbpA and hgbB: iron acquisition) by PCR and sequencing and submitted to multilocus sequence typing (MLST).ResultsThe eight PmA strains were classified as follows: five as highly pathogenic (HP) for causing necrotic bronchopneumonia and diffuse fibrinous pleuritis and pericarditis; one as low pathogenic for causing only focal bronchopneumonia; and two as nonpathogenic because they did not cause injury to any pig. PCR for the gene pfhA was positive for all five HP isolates. Sequencing demonstrated that the pfhA region of the HP strains comprised four genes: tpsB1, pfhA1, tpsB2 and pfhA2. The low and nonpathogenic strains did not contain the genes tpsB2 and pfhA2. A deletion of four bases was observed in the pfhA gene in the low pathogenic strain, and an insertion of 37 kb of phage DNA was observed in the nonpathogenic strains. MLST clustered the HP isolates in one group and the low and nonpathogenic isolates in another. Only the nonpathogenic isolates matched sequence type 10; the other isolates did not match any type available in the MLST database.ConclusionsThe hypothesis that some PmA strains are primary pathogens and cause disease in pigs without any co-factor was confirmed. The pfhA region, comprising the genes tpsB1, tpsB2, pfhA1 and pfhA2, is related to the pathogenicity of PmA. The HP strains can cause necrotic bronchopneumonia, fibrinous pleuritis and pericarditis in pigs and can be identified by PCR amplification of the gene pfhA2.

Clinical characteristics and prognostic factors of pneumonia in patients with and without rheumatoid arthritis.

BackgroundTo elucidate the characteristics of pneumonia in rheumatoid arthritis (RA) patients and to assess whether pneumonia in RA patients differs from that in non-RA patients.MethodsWe retrospectively divided pneumonia patients into two groups, those with RA and those without RA, and compared the two groups. We evaluated the risk factors for mortality with univariate and multivariate logistic regression analysis.ResultsAmong 1549 patients, 71 had RA. The RA patients with pneumonia were 71.0±8.9 years old, 54.9% were female, 40.9% had a smoking history, and 71.8% had underlying respiratory disease. Female sex, non-smoker, and respiratory comorbidities were statistically more frequent in the RA patients than non-RA patients. The most frequent causative microbial agents of pneumonia in the RA patients were Streptococcus pneumoniae, Pseudomonas aeruginosa, Haemophilus influenzae, Mycoplasma pneumoniae, and influenza virus, whereas those of pneumonia in non-RA patients were S. pneumoniae, influenza virus, M. pneumoniae, Legionella spp., P. aeruginosa, H. influenzae, and Moraxella catarrhalis. Polymicrobial infection were identified as etiologies more frequently in the RA patients than non-RA patients. Although the severity of pneumonia did not differ between the two groups, mortality was statistically higher in the RA patients than non-RA patients. Multivariate analysis showed RA to be an independent risk factor for mortality.ConclusionsP. aeruginosa, H. influenzae, M. catarrhalis, and polymicrobial infection were statistically more frequent etiologies of pneumonia in the RA patients than non-RA patients. RA itself was found to be an independent risk factor for mortality from pneumonia.

Pasteurella multocida inactivated with ferric chloride and adjuvanted with bacterial DNA is a potent and efficacious vaccine in Balb/c mice.

Pasteurella multocida (P. multocida) is a principal pathogen of domestic animals and an opportunistic pathogen of humans. It is the causative agent of pneumonia and haemorrhagic septicaemia in cattle, sheep and goats, fowl cholera in chickens and progressive atrophic rhinitis in swine. In this study, we investigated the humoral and cellular immune responses and protective immunity conferred by an iron-inactivated vaccine with bacterial DNA (IIV+bDNA) as an adjuvant in mice. P. multocida was grown in BHI broth, inactivated with formalin and FeCl3 and adjuvanted with alum and bDNA. Mice were immunized with two whole-cell inactivated vaccine doses 2 weeks apart. The animals were challenged 4 weeks after booster immunization. Immunogens (vaccines and bDNA) posed no safety problems when mice were injected subcutaneously (s/c) with these preparations. The serum antibody titres were tested by ELISA. At 28 days post immunization, cell-mediated immunity responses were determined. The responses were measured by assay of IL-6 and IL-12 in lymphocyte spleen culture supernatants. ELISA results showed that the levels of antibodies in iron inactivated with bDNA adjuvant groups were higher than in the formalin inactivated with alum adjuvant vaccine group. The protection rate of IIV+bDNA adjuvant vaccine was superior to that of the other vaccines and it protected 100 % of the challenge group mice. Following immunization, bDNA promoted increased production of interleukins compared to the control groups. These studies indicate that bDNA is effective as an immune adjuvant, and along with stimulatory bDNA represent promising new humoral and cellular immune enhancers for vaccination applications. In addition, this vaccine is able to provide long-term protection against infection.

The Relationship Between CMV Seropositivity and Fungal Pneumonia in Renal Transplant Patients

Introduction Invasive fungal infections are major problem in solid organ transplant (SOT) recipients because of their net state of immuno-suppression. Overall, the most common fungal infection in SOT is candidiasis, followed by aspergillosis and cryptococcosis. Cytomegalovirus (CMV) is the most common viral infection following renal transplant (RT). About one-quarter of renal transplant recipients developed Seropositivity (CMVSP). CMSP is also described as a risk factor for fungal pneumonia because it led to a pronounced cellular immunosuppression in the SOT patients. To date, the role of CMVSP on the development of fungal pneumonia in SOT patients has not been investigated so far. In this study, we aimed to investigate the role of CMVSP on the development of Aspergillus pneumonia in renal transplant patients retrospectively. Materials and Methods We evaluated a total of 401 patients (male/female=273/128) who underwent renal transplant between 2011 and 2017. Medical records of these patients were reviewed for demographic, clinical, radiographic, laboratory and microbiology data, laboratory method for determining CMV. The diagnosis of pneumonia based on clinical respiratory symptoms and signs, imaging findings (chest radiography and/or computed tomography), positive microbiological tests and laboratory findings. Immune suppressive drugs and clinical outcome data were also noted. Results Our study included in 153 (38%) (male/female= 97/55 and with the mean age of 35 ± 14 years) renal transplantation patients (44 cadaveric, 109 living donors) who had pneumonia in our study population. The most frequently determined immunosuppression therapy included prednisolone plus cyclosporine, tacrolimus, or rapamycin. CMVSP was found in 77 (%0.3) of the patients. Twenty-one of the patients had positive culture from respiratory specimens. Fungal agents were detected in 16 (77%) of the patients and Aspergillus fumigatus was the most common (38%) fungus species. No statistical significant correlation was found between Seropositivity and microbiological agents of pneumonia (p>0.05) in our study group. Conclusions Our results suggest that Seropositivity may not be a risk for fungal pneumonia in RT patients. Further studies needed to reveal whether CMVSP is a risk for fungal pneumonia or not in larger fungal pneumonia series in RT patients.

Application of BioFire FilmArray Blood Culture Identification panel for rapid identification of the causative agents of ventilator-associated pneumonia

ObjectiveTo evaluate the ability of the BioFire FilmArray Blood Culture Identification (BCID) panel to rapidly detect pathogens producing late-onset ventilator-associated pneumonia (VAP), a severe infection often produced by Gram-negative bacteria. These microorganisms are frequently multidrug resistant and typically require broad-spectrum empiric treatment. MethodsIn the context of an international multicentre clinical trial (MagicBullet), respiratory samples were collected at the time of suspicion of VAP from 165 patients in 32 participating hospitals in Spain, Greece and Italy. Microorganisms were identified using the BCID panel and compared with results obtained by conventional microbiologic techniques. ResultsPseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae were the most commonly identified species, representing 54.7% (70/128) of microorganisms. The BCID panel showed high global specificity (98.1%; 95% confidence interval, 96–100) and negative predictive values (96.6%) and a global sensitivity and positive predictive value of 78.6% (95% confidence interval, 70–88) and 87.3%, respectively, for these microorganisms. Importantly, the BCID panel provided results in only 1 hour directly from respiratory samples with minimal sample processing times. ConclusionsThe BCID panel may have clinical utility in rapidly ruling out microorganisms causing VAP, specifically multidrug-resistant Gram-negative species. This could facilitate the optimization of empiric treatment.

Prolonged antibiotic prophylaxis after thoracoabdominal esophagectomy does not reduce the risk of pneumonia in the first 30days: a retrospective before-and-after analysis.

Thoracoabdominal esophageal resection for malignant disease is frequently associated with pulmonary infection. Whether prolonged antibiotic prophylaxis beyond a single perioperative dose is advantageous in preventing pulmonary infection after thoracoabdominal esophagectomy remains unclear. In this retrospective before-and-after analysis, 173 patients between January 2009 and December 2014 from a prospectively maintained database were included. We evaluated the effect of a 5-day postoperative course of moxifloxacin, which is a frequently used antimicrobial agent for pneumonia, on the incidence of pulmonary infection and mortality after thoracoabdominal esophagectomy. 104 patients received only perioperative antimicrobial prophylaxis (control group) and 69 additionally received a 5-day postoperative antibiotic therapy with moxifloxacin (prolonged-course). 22 (12.7%) of all patients developed pneumonia within the first 30days after surgery. No statistically significant differences were seen between the prolonged group and control group in terms of pneumonia after 7 (p = 0.169) or 30days (p = 0.133), detected bacterial species (all p > 0.291) and 30-day mortality (5.8 vs 10.6%, p = 0.274). A preemptive 5-day postoperative course of moxifloxacin does not reduce the incidence of pulmonary infection and does not improve mortality after thoracoabdominal esophagectomy.

Development of a rapid isolation and culture method of wild type Mycoplasma spp. in pigs

Mycoplasma spp. are extracellular bacteria that colonize on the respiratory epithelium of humans and animals. It is a causative agent of pneumonia commonly complicated by opportunistic infectious bacteria. Mycoplasma spp. infection cause relatively mild disease in the absence of environmental stressors, but when complicated by secondary bacterial invaders the resultant disease can cause obvious clinical disease and severe production losses in intensively reared pigs Mycoplasma spp. are highly fastidious bacteria, difficult to culture and slow growing. Many species of Mycoplasma spp. are important pathogens causing respiratory infection in animals and known to induce huge economic losses. The aims of the present study were to develop a rapid isolation and culture method of wild type Mycoplasma spp. in pigs. We used Mycoplasma spp. genus specific direct PCR without DNA extraction procedure using PhireⓇ Animal Tissue Direct PCR Kit from the lung tissues with pneumonia lesions. Therefore, we could save the time for tissue processing and increase the accuracy of Mycoplasma spp. inclusion prediction in lung tissues. Thereafter, we used the optimized media to isolate and culture Mycoplasma spp. As the results, Mycoplasma spp. could be isolated and cultured quickly and efficiently. These results could provide an efficient strategy and method for the rapid and accurate isolation and culture of wild type Mycoplasma spp. in pigs.