Characterization of nonencapsulated Actinobacillus pleuropneumoniae serovar K12:O3 isolates.

Three Actinobacillus pleuropneumoniae isolates from clinical cases of porcine pleuropneumonia were positive by capsular serovar 12-specific PCR assay, but not reactive to antiserum prepared against serovar 12 using the rapid slide agglutination (RSA) test. The isolates were positive for apxIICA, apxIIICA, apxIBD, apxIIIBD, and apxIVA in the PCR toxin gene assay, which is the profile seen in serovars 2, 4, 6, 8, and 15, and reacted with antisera against serovars 3, 6, 8, 15, and 17. Nucleotide sequence analysis revealed that genes involved in the biosynthesis of capsular polysaccharide of the 3 isolates were identical or nearly identical to those of serovar 12. However, genes involved in the biosynthesis of O-polysaccharide of the 3 isolates were highly similar to those of reference strains of serovars 3, 6, 8, 15, 17, and 19. In agreement with results from the RSA test, transmission electron microscopic analysis confirmed the absence of detectable capsular material in the 3 isolates. The existence of nonencapsulated A. pleuropneumoniae serovar K12:O3 would hamper precise serodetection.

Rapid detection of Actinobacillus pleuropneumoniae targeting the apxIVA gene for diagnosis of contagious porcine pleuropneumonia in pigs by polymerase spiral reaction.

Actinobacillus pleuropneumoniae is the primary aetiological agent of contagious porcine pleuropneumonia associated with serious economic impact on pig husbandry worldwide. Diagnosis of the disease by existing techniques including isolation and identification of bacteria followed by serotyping, serological techniques, conventional PCR, real-time PCR and LAMP assays are cumbersome, time-consuming, costly and not suitable for rapid field application. A novel isothermal polymerase chain reaction (PSR) technique is standardized for all the reagents, incubation time and incubation temperature against A. pleuropneumoniae. The sensitivity of the assay was determined against various dilutions of purified DNA and total bacterial count. The specificity of the assay was determined against 11 closely related bacterial isolates. The relative sensitivity and specificity were compared with bacterial isolation, conventional PCR and real-time PCR assays. The PSR assay for specific detection was standardized at 64°C for 30 min of incubation in a water bath. The result was visible by the naked eye after centrifugation of the reaction mixture or after incorporation of SYBR Green dye as yellowish-green fluorescence. The technique was found to be 100% specific and equally sensitive with real-time PCR and 10 times more sensitive than conventional PCR. The PSR assay could be applicable in the detection of the organisms in porcine nasal swabs spiked with A. pleuropneumoniae. This is the first-ever report on the development of PSR for specific detection of A. pleuropneumoniae and can be applied for early diagnosis at the field level.

Novel DNA Markers for Identification of Actinobacillus pleuropneumoniae.

Actinobacillus pleuropneumoniae causes porcine pleuropneumonia, an important disease in the pig industry. Accurate and sensitive diagnostics such as DNA-based diagnostics are essential for preventing or responding to an outbreak. The specificity of DNA-based diagnostics depends on species-specific markers. Previously, an insertion element was found within an A. pleuropneumoniae-specific gene commonly used for A. pleuropneumoniae detection, prompting the need for additional species-specific markers. Herein, 12 marker candidates highly conserved (99 - 100% identity) among 34 A. pleuropneumoniae genomes (covering 13 serovars) were identified to be A. pleuropneumoniae-specific in silico, as these sequences are distinct from 30 genomes of 13 other Actinobacillus and problematic [Actinobacillus] species and more than 1700 genomes of other bacteria in the Pasteurellaceae family. Five marker candidates are within the apxIVA gene, a known A. pleuropneumoniae-specific gene, validating our in silico marker discovery method. Seven other A. pleuropneumoniae-specific marker candidates within the eamA, nusG, sppA, xerD, ybbN, ycfL, and ychJ genes were validated by polymerase chain reaction (PCR) to be specific to 129 isolates of A. pleuropneumoniae (covering all 19 serovars), but not to four closely related Actinobacillus species, four [Actinobacillus] species, or seven other bacterial species. This is the first study to identify A. pleuropneumoniae-specific markers through genome mining. Seven novel A. pleuropneumoniae-specific DNA markers were identified by a combination of in silico and molecular methods and can serve as additional or alternative targets for A. pleuropneumoniae diagnostics, potentially leading to better control of the disease. IMPORTANCE Species-specific markers are crucial for infectious disease diagnostics. Mutations within a marker sequence can lead to false-negative results, inappropriate treatment, and economic loss. The availability of several species-specific markers is therefore desirable. In this study, 12 DNA markers specific to A. pleuropneumoniae, a pig pathogen, were simultaneously identified. Five marker candidates are within a known A. pleuropneumoniae-specific gene. Seven novel markers can be used as additional targets in DNA-based diagnostics, which in turn can expedite disease diagnosis, assist farm management, and lead to better animal health and food security. The marker discovery strategy outlined herein requires less time, effort, and cost, and results in more markers compared with conventional methods. Identification of species-specific markers of other pathogens and corresponding infectious disease diagnostics are possible, conceivably improving health care and the economy.

Severe bilateral pleuropneumonia caused by Legionella sainthelensi: a case report.

BACKGROUND: Legionella spp. are ubiquitous freshwater bacteria responsible for rare but potentially severe cases of Legionnaires' disease (LD). Legionella sainthelensi is a non-pneumophila Legionella species that was first isolated in 1980 from water near Mt. St-Helens (USA). Although rare cases of LD caused by L. sainthelensi have been reported, very little data is available on this pathogen. CASE PRESENTATION: We describe the first documented case of severe bilateral pleuropneumonia caused by L. sainthelensi. The patient was a 35-year-old woman with Sharp's syndrome treated with long-term hydroxychloroquine and corticosteroids who was hospitalized for an infectious illness in a university hospital in Reunion Island (France). The patient's clinical presentation was complicated at first (bilateral pneumonia, multiloculated pleural effusion, then bronchopleural fistula) but her clinical condition eventually improved with the reintroduction of macrolides (spiramycin) in intensive care unit. Etiological diagnosis was confirmed by PCR syndromic assay and culture on bronchoalveolar lavage. CONCLUSIONS: To date, only 14 documented cases of L. sainthelensi infection have been described worldwide. This pathogen is difficult to identify because it is not or poorly detected by urinary antigen and molecular methods (like PCR syndromic assays that primarily target L. pneumophila and that have only recently been deployed in microbiology laboratories). Pneumonia caused by L. sainthelensi is likely underdiagnosed as a result. Clinicians should consider the possibility of non-pneumophila Legionella infection in patients with a compatible clinical presentation when microbiological diagnostic tools targeted L. pneumophila tested negative.

Pleuropneumonia resulting from varicella and COVID-19 co-infection in a 10-month-old infant.

COVID-19 is a new disease leading to respiratory complications in adults. Children appear to have more modest symptoms than adults. Varicella is often described as a benign disease in the pediatric population. However, patients with varicella and COVID-19 co-infection can develop a more serious respiratory infection. We report the case of an infant who had a co-infection with both viruses that led to pleuropneumonia. The main question in the present case concerns the link between COVID-19 and varicella infection, and the possible modulation in immune response due to the two virus infections.

Characterization of nontypeable Actinobacillus pleuropneumoniae isolates.

Two Actinobacillus pleuropneumoniae isolates from clinical cases of porcine pleuropneumonia in Japan were positive in the capsular serovar 15-specific PCR assay, but nontypeable (NT) in the agar gel precipitation (AGP) test. Nucleotide sequence analysis of gene clusters involved in the biosynthesis of capsular polysaccharide (CPS) and lipopolysaccharide O-polysaccharide (O-PS) revealed that both clusters contained transposable element ISApl1 of A. pleuropneumoniae belonging to the IS30 family. Immunoblot analysis revealed that these 2 isolates could not produce O-PS. We conclude that the ISApl1 of A. pleuropneumoniae can interfere in the biosynthesis of both CPS and O-PS.

Contagious Bovine Pleuropneumonia: A Comprehensive Overview.

Contagious bovine pleuropneumonia (CBPP) is a respiratory disease of cattle that is listed as notifiable by the World Organization for Animal Health. It is endemic in sub-Saharan Africa and causes important productivity losses due to the high mortality and morbidity rates. CBPP is caused by Mycoplasma mycoides subsp. mycoides (Mmm) and is characterized by severe fibrinous bronchopneumonia and pleural effusion during the acute to subacute stages and by pulmonary sequestra in chronic cases. Additional lesions can be detected in the kidneys and in the carpal and tarsal joints of calves. Mmm infection occurs through the inhalation of infected aerosol droplets. After the colonization of bronchioles and alveoli, Mmm invades blood and lymphatic vessels and causes vasculitis. Moreover, Mmm can be occasionally demonstrated in blood and in a variety of other tissues. In the lung, Mmm antigen is commonly detected on bronchiolar and alveolar epithelial cells, in lung phagocytic cells, within the wall of blood and lymphatic vessels, inside necrotic areas, and within tertiary lymphoid follicles. Mmm antigen can also be present in the cytoplasm of macrophages within lymph node sinuses, in the germinal center of lymphoid follicles, in glomerular endothelial cells, and in renal tubules. A complete pathological examination is of great value for a rapid presumptive diagnosis, but laboratory investigations are mandatory for definitive diagnosis. The purpose of this review is to describe the main features of CBPP including the causative agent, history, geographic distribution, epidemiology, clinical course, diagnosis, and control. A special focus is placed on gross and microscopic lesions in order to familiarize veterinarians with the pathology and pathogenesis of CBPP.

Glomerulonefritis infecciosa y pleuroneumonía por Streptococcus pneumoniae. Caso clínico pediátrico / Infectious glomerulonephritis and pleuropneumonia due to Streptococcus pneumoniae. Pediatric clinical case

La glomerulonefritis aguda desencadenada por Streptococcus pneumoniae es una patología de baja prevalencia. Existen diversos reportes que comunican distintas cepas nefritogénicas; sin embargo, la 6C ha sido escasamente señalada como tal.Se presenta el caso de un paciente de 4 años, quien ingresó a Terapia Intensiva con pleuroneumonía por Streptococcus pneumoniae serotipo 6C y desarrolló, de modo concomitante, edemas, hipertensión arterial, hematuria, proteinuria, disminución del filtrado glomerular y del nivel de complemento C3. Se diagnosticó glomerulonefritis aguda. Su evolución fue satisfactoria en un breve plazo. Esta patología, por lo general, es de curso transitorio y benigno; sin embargo, en ocasiones, puede complicar la evolución de un paciente críticamente enfermo, por lo cual se hace necesario tenerla entre los diagnósticos diferenciales para considerar.

Acute glomerulonephritis caused by Streptococcuspneumoniaeis a low prevalence pathology. There are several reports communicating different nephritogenic serotypes, however, 6C has been scarcely indicated as such. It is presented the case of a 4-year-old patient who entered Intensive Therapy Unit with pleuropneumonia due to Streptococcuspneumoniae serotype 6C and concomitantly developed edemas, arterial hypertension, hematuria, proteinuria, decreased glomerular filtration rate and C3 complement level. Acute glomerulonephritis was diagnosed. His evolution was satisfactory in a short time. This pathology is usually of a transitory and benign course; however, sometimes it can potentially complicate the evolution of a critically ill patient, so it is necessary to have it among the differential diagnoses to consider.

[Infectious glomerulonephritis and pleuropneumonia due to Streptococcus pneumoniae. Pediatric clinical case]. / Glomerulonefritis infecciosa y pleuroneumonía por Streptococcus pneumoniae. Caso clínico pediátrico.

Acute glomerulonephritis caused by Streptococcus pneumoniae is a low prevalence pathology. There are several reports communicating different nephritogenic serotypes, however, 6C has been scarcely indicated as such. It is presented the case of a 4-year-old patient who entered Intensive Therapy Unit with pleuropneumonia due to Streptococcus pneumoniae serotype 6C and concomitantly developed edemas, arterial hypertension, hematuria, proteinuria, decreased glomerular filtration rate and C3 complement level. Acute glomerulonephritis was diagnosed. His evolution was satisfactory in a short time. This pathology is usually of a transitory and benign course; however, sometimes it can potentially complicate the evolution of a critically ill patient, so it is necessary to have it among the differential diagnoses to consider.

La glomerulonefritis aguda desencadenada por Streptococcus pneumoniae es una patología de baja prevalencia. Existen diversos reportes que comunican distintas cepas nefritogénicas; sin embargo, la 6C ha sido escasamente señalada como tal. Se presenta el caso de un paciente de 4 años, quien ingresó a Terapia Intensiva con pleuroneumonía por Streptococcus pneumoniae serotipo 6C y desarrolló, de modo concomitante, edemas, hipertensión arterial, hematuria, proteinuria, disminución del filtrado glomerular y del nivel de complemento C3. Se diagnosticó glomerulonefritis aguda. Su evolución fue satisfactoria en un breve plazo. Esta patología, por lo general, es de curso transitorio y benigno; sin embargo, en ocasiones, puede complicar la evolución de un paciente críticamente enfermo, por lo cual se hace necesario tenerla entre los diagnósticos diferenciales para considerar.

Contagious caprine pleuropneumonia - a comprehensive review.

Contagious caprine pleuropneumonia (CCPP) is a serious disease of goats, occasionally sheep and wild ruminants, caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). The disease is characterized by severe serofibrinous pleuropneumonia, very high morbidity (∼100%), and mortality (80-100%). CCPP affects goats in more than 40 countries of the world thereby posing a serious threat to goat farming around the globe. The characteristic clinical signs of CCPP are severe respiratory distress associated with sero-mucoid nasal discharge, coughing, dyspnea, pyrexia, pleurodynia, and general malaise. In later stages, severe lobar fibrinous pleuropneumonia, profuse fluid accumulation in pleural cavity, severe congestion of lungs and adhesion formation is observed. Mycoplasmal antigen interactions with host immune system and its role in CCPP pathogenesis are not clearly understood. CCPP is not a zoonotic disease. Diagnosis has overcome cumbersome and lengthy conventional tests involving culture, isolation, and identification by advanced serological (LAT, cELISA) or gene-based amplification of DNA (PCR, RFLP, and hybridization) and sequencing. The latex agglutination test (LAT) is rapid, simple, and better test for field and real-time diagnosis applicable to whole blood or serum and is more sensitive than the CFT and easier than the cELISA. Moreover, the studies on antibiotic sensitivity and exploration of novel antibiotics (fluoroquinolones, macrolides) can help in better therapeutic management besides preventing menace of antibiotic resistance. Re-visiting conventional prophylactic measures focussing on developing novel strain-based or recombinant vaccines using specific antigens (capsular or cellular) should be the most important strategy for controlling the disease worldwide.

Pleuroparenchymal fibroelastosis with positive MPO-ANCA diagnosed with a CT-guided percutaneous needle biopsy.

A 67-year-old woman was referred to our hospital because of gradually increasing dyspnoea on exertion for 6 months. Chest CT scan showed subpleural parenchymal fibrotic opacities with traction bronchiectasis in the bilateral upper lung fields. Serum rheumatoid factor and myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA) were positive. There was no evident reason to suspect connective tissue disease such as ANCA-associated vasculitis or rheumatoid arthritis. We performed a CT-guided percutaneous needle biopsy of the subpleural lesion that showed slight uptake on the fluorodeoxyglucose-positron emission tomography (FDG-PET) CT scan. This specimen showed subpleural fibrosis as evidenced by an abnormal increase of elastic tissue and minimal collagen deposition, which indicated pleuroparenchymal fibroelastosis (PPFE). Although PPFE can be associated with a variety of causes, its association with MPO-ANCA is unknown. A CT-guided transthoracic lung biopsy caused no adverse events and was useful in the diagnosis of PPFE in our patient.

Isolation and molecular characterization of a urease-negative Actinobacillus pleuropneumoniae mutant.

An atypical urease-negative mutant of Actinobacillus pleuropneumoniae serovar 2 was isolated in Japan. Nucleotide sequence analysis of the urease gene cluster revealed that the insertion of a short DNA sequence into the cbiM gene was responsible for the urease-negative activity of the mutant. Veterinary diagnostic laboratories should be watchful for the presence of aberrant urease-negative A. pleuropneumoniae isolates.

The value of molecular techniques to diagnose Ureaplasma urealyticum and Nocardia farcinica pleuropneumonia in a patient with diffuse large B-cell lymphoma.

An unusual case of pleural empyema related to Nocardia farcinica and Ureaplasma urealyticum, occurring after autologous haematopoietic stem cell transplantation in a 30-year-old patient with lymphoma, is reported. This case illustrates the role of repeated and comprehensive microbiological investigations and the contribution of molecular techniques in reaching the aetiological diagnosis.

Thoracic splenosis mimicking a pleuropneumonia: A case report.

RATIONALE: Splenosis is the development of one or more heterotopic splenic tissue autoimplants following rupture of the spleen and remains mostly asymptomatic. PATIENT CONCERNS: We report a case of a 50-year old post-traumatic splenectomized man admitted for a left side community acquired pneumonia resistant to antibiotics. DIAGNOSES: The diagnosis of intrathoracic ectopic spleen was suspected because of the history of spleen trauma with diaphragm rupture and the absence of Howell-Jolly bodies. INTERVENTIONS: Technetium (Tc)-m colloid scintigraphy SPECT, fused with CT scan showed an intense radionuclide uptake on hyper vascularized masses without any additional pathologic uptake and confirmed the diagnosis of thoracic splenosis. OUTCOMES: Despite any lifelong penicillin prophylaxis, he had no history of infections eight years after the diagnosis. LESSONS: Physician must be aware of this differential diagnosis and of its consequences. Depending on its size and location, it may lead to incorrect diagnosis (tumor, empyema, abscess ...), treatment and invasive procedures while the diagnosis of splenosis only relies upon imaging studies associated with functionnal study of the uptake of particles or cells.

Nucleotide sequence analysis of a DNA region involved in capsular polysaccharide biosynthesis reveals the molecular basis of the nontypeability of two Actinobacillus pleuropneumoniae isolates.

The aim of our study was to reveal the molecular basis of the serologic nontypeability of 2 Actinobacillus pleuropneumoniae field isolates. Nine field strains of A. pleuropneumoniae, the causative agent of porcine pleuropneumonia, were isolated from pigs raised on the same farm and sent to our diagnostic laboratory for serotyping. Seven of the 9 strains were identified as serovar 15 strains by immunodiffusion tests. However, 2 strains, designated FH24-2 and FH24-5, could not be serotyped with antiserum prepared against serovars 1-15. Strain FH24-5 showed positive results in 2 serovar 15-specific PCR tests, whereas strain FH24-2 was only positive in 1 of the 2 PCR tests. The nucleotide sequence analysis of gene clusters involved in capsular polysaccharide biosynthesis of the 2 nontypeable strains revealed that both had been rendered nontypeable by the action of ISApl1, a transposable element of A. pleuropneumoniae belonging to the IS30 family. The results showed that ISApl1 of A. pleuropneumoniae can interfere with both the serologic and molecular typing methods, and that nucleotide sequence analysis across the capsular gene clusters is the best means of determining the cause of serologic nontypeability in A. pleuropneumoniae.

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Isolation and genetic characterization of an Actinobacillus pleuropneumoniae serovar K12:O3 strain.

An atypical Actinobacillus pleuropneumoniae serovar 12 strain, termed QAS106, was isolated from a clinical case of porcine pleuropneumonia in Japan. An immunodiffusion (ID) test identified the strain as serovar 12. However, the ID test also demonstrated that strain QAS106 shared antigenic determinants with both the serovar 3 and 15 reference strains. Strain QAS106 was positive in the capsular serovar 12-specific polymerase chain reaction (PCR) assay, while the PCR toxin gene profiling and omlA PCR typing assays indicated that strain QAS106 was similar to serovar 3. The nucleotide sequence of the 16S ribosomal DNA (rDNA) of strain QAS106 was identical with that of serovars 3 and 12, but it showed 99.7% identity with that of serovar 15. Nucleotide sequence analysis revealed that genes involved in biosynthesis of the capsular polysaccharide (CPS) of strain QAS106 were identical to those of serovar 12 at the amino acid level. On the other hand, strain QAS106 would express putative proteins involved in the biosynthesis of lipopolysaccharide (LPS) O-polysaccharide (O-PS), the amino acid sequences of which were identical or nearly identical to those of serovars 3 and 15. In conclusion, strain QAS106 should be recognized as K12:O3, even though typical serovar 12 strains are K12:O12. The emergence of an atypical A. pleuropneumoniae serovar 12 strain expressing a rare combination of CPS and O-PS antigens would hamper precise serodiagnosis by the use of either CPS- or LPS-based serodiagnostic methodology alone.

[Pleuropulmonary tularemia in a 63-year-old hunter in Germany]. / Pleuropulmonale Tularämie bei einem 63-jährigen Jäger in Deutschland.

HISTORY: A 63-year-old amateur hunter without relevant preexisting diseases presented with cough lasting for 4 weeks and B symptoms. INVESTIGATIONS: Radiography and ultrasonography showed a left-sided pleural effusion. Laboratory markers of infection were in normal range. A thoracocentesis was performed for diagnostic purposes. TREATMENT AND COURSE: Culture of Francisella tularensis spp. holarctica from pleural fluid led to the diagnosis of tularemia with unusual manifestation as an isolated one-sided pleuritis, confirmed by highly positive serology. The patient fully recovered on oral treatment with doxycycline. CONCLUSION: Due to its low prevalence in Germany and multiple possible clinical manifestations, the diagnosis of tularemia can be difficult. Hence, it is decisive to include tularemia in the differential diagnosis, especially if anamnestic or epidemiological evidence exists.

Chest pain in primary care: is the localization of pain diagnostically helpful in the critical evaluation of patients?--A cross sectional study.

BACKGROUND: Chest pain is a common complaint and reason for consultation in primary care. Traditional textbooks still assign pain localization a certain discriminative role in the differential diagnosis of chest pain. The aim of our study was to synthesize pain drawings from a large sample of chest pain patients and to examine whether pain localizations differ for different underlying etiologies. METHODS: We conducted a cross-sectional study including 1212 consecutive patients with chest pain recruited in 74 primary care offices in Germany. Primary care providers (PCPs) marked pain localization and radiation of each patient on a pictogram. After 6 months, an independent interdisciplinary reference panel reviewed clinical data of every patient, deciding on the etiology of chest pain at the time of patient recruitment. PCP drawings were entered in a specially designed computer program to produce merged pain charts for different etiologies. Dissimilarities between individual pain localizations and differences on the level of diagnostic groups were analyzed using the Hausdorff distance and the C-index. RESULTS: Pain location in patients with coronary heart disease (CHD) did not differ from the combined group of all other patients, including patients with chest wall syndrome (CWS), gastro-esophageal reflux disease (GERD) or psychogenic chest pain. There was also no difference in chest pain location between male and female CHD patients. CONCLUSIONS: Pain localization is not helpful in discriminating CHD from other common chest pain etiologies.