Non-tuberculosis Mycobacteria Research Articles

The natural route of infection of guinea pigs from cows with tuberculosis in a cattle farm

The success of the fight against tuberculosis depends on timely and accurate diagnosis, since the identification and removal of animals suspected of the disease from the herd at the very beginning of the development of the infectious process is the main condition for recreational activities. Prevention of tuberculosis is based on the implementation of measures to prevent the introduction of the pathogen into a safe economy. Control of well-being is achieved by checking the number of cattle by intradermal tuberculin test twice a year (in spring and autumn), young cattle from two months of age and cattle of the feeding group - once a year. Carrying out such studies is quite sufficient, since it is known that the delayed-type hypersensitivity state develops 7-45 days after infection of cattle with M. bovis tuberculosis pathogen. The main disadvantage of allergic studies is the well-known fact that the reaction to the intradermal administration of tuberculin indicates only the sensitization of the animal's body by various types of mycobacteria. Non-tuberculosis mycobacteria do not cause pathological changes characteristic of tuberculosis in cattle and even in guinea pigs that are very sensitive to infection. Therefore, when monitoring the welfare of farms, the results of allergic studies are considered indicative, and the reacting animals are suspected of being infected. Decisive in the diagnosis is pathological or laboratory confirmation of tuberculosis in animals killed for diagnostic purposes. There is little data in the literature on the timing of the development of the infectious process and the identification of pathological changes in cattle characteristic of tuberculosis. Data on the timing of the detection of changes characteristic of tuberculosis in guinea pigs with natural infection from sick cows was not found. Previously conducted studies in an experiment on natural infection of cattle with tuberculosis, the authors found that when healthy cows are introduced into a dysfunctional herd, they become infected with tuberculosis with the formation of characteristic pathological changes approximately 12 months after they are kept together with sick animals. In an experiment on natural infection of guinea pigs, the authors found that guinea pigs in the same room with cows infected with M. bovis become infected and develop tuberculosis. Changes characteristic of tuberculosis in experimental guinea pigs are detected 5 months after they are kept together with sick cows. Based on a review of the literature and the results of the studies conducted, the authors propose to use guinea pigs as a laboratory model for reproducing tuberculosis in order to control the welfare of livestock farms in safe farms or to assess the quality of recreational activities in farms with tuberculosis.

Direct Molecular Characterization of Acid-Fast Bacilli Smear of Nontuberculosis Mycobacterium Species Causing Pulmonary Tuberculosis in Guna Yala Region, Panama.

Mycobacterium tuberculosis (MTB) stands out as the main causative agent of pulmonary tuberculosis (TB). However, nontuberculous mycobacteria (NTM) species also have the potential to infect and cause TB in susceptible individuals. The objective of this study was to identify NTM species that cause public health problems in remote areas. The study was carried out using 105 sputum smears obtained from patients from the Guna Yala Region of Panama with clinical signs suggestive of TB. DNA was extracted from sputum smears. Nontuberculous mycobacteria and MTB were characterized using polymerase chain reaction restriction analysis (hsp65, rpob) and an evaluation of 24-mycobacterial interspersed repetitive units-variable number of tandem repeats loci. Twenty-six Mycobacterium species were characterized; 19 (18%) were identified as MTB, and 7 (6.7%) were identified as NTM (four M. avium complex, two M. haemophilum, one M. tusciae). These results suggest that at least one in five cases of pulmonary TB among this population is caused by an NTM. Thus, identifying the bacteria causing pulmonary disease is key even in remote regions of the world where standard diagnosis and culture are not available. Strengthening the laboratory capacity within the Guna Yala Region is needed to identify NTM infections promptly.

Clinical course of peritoneal dialysis-related peritonitis due to non-tuberculosis mycobacterium – A single centre experience spanning 20 years

Peritoneal dialysis (PD)-related peritonitis caused by non-tuberculous mycobacteria (NTM) are difficult to diagnose, is associated with significant morbidity and mortality, and clinical course remains unclear. We determined the prevalence and clinical course of peritonitis caused by these organisms through our kidney registry over 20-year period. We reviewed all patients with NTM peritonitis identified in our tertiary centre between July 2000 and July 2020. The demographic characteristics, microbiological and clinical outcomes were examined. Among 27 patients identified, 20 patients presented with abdominal pain and all had cloudy peritoneal fluid. Twenty-one cases had concomitant exit site infection and 14 cases had prior antibiotic use. The majority of the cases are caused by Mycobacterium chelonae (37%) and Mycobacterium fortuitum (29.7%), with most being resistant to fluoroquinolones (59.3%) and cefoxitin (73.1%). They are all sensitive to amikacin otherwise. None of the cases achieve primary response at day 10 and 20 cases resulted in Tenckhoff catheter removal. Only two of them were able to resume PD. Eight patients died in our cohort. The presence of exit site infection, the use of prior antibiotics and topical disinfectants did not associate with a poorer outcome. NTM peritonitis remains difficult to treat and often with a delay in diagnosis. Refractory peritonitis with negative culture and a poor response to standard antibiotics should raise a possibility of NTM infection and prompt catheter removal and an expert with experience treating NTM infections should be consulted.

Detecting Mycobacterium tuberculosis complex and rifampicin resistance via a new rapid multienzyme isothermal point mutation assay

Simple, rapid, and accurate detection of the Mycobacterium tuberculosis complex (MTBC) and drug resistance is critical for improving patient care and decreasing the spread of tuberculosis. To this end, we have developed a new simple and rapid molecular method, which combines multienzyme isothermal rapid amplification and a lateral flow strip, to detect MTBC and simultaneously detect rifampin (RIF) resistance. Our findings showed that it has sufficient sensitivity and specificity for discriminating 118 MTBC strains from 51 non-tuberculosis mycobacteria strains and 11 of the most common respiratory tract bacteria. Further, compared to drug susceptibility testing, the assay has a sensitivity, specificity, and accuracy of 54.1%, 100.0%, and 75.2%, respectively, for detection of RIF resistance. Some of the advantages of this assay are that no special instrumentation is required, a constant low temperature of 39 °C is sufficient for the reaction, the turnaround time is less than 20 min from the start of the reaction to read out and the result can be seen with the naked eye and does not require specialized training. These characteristics of the new assay make it particularly useful for detecting MTBC and RIF resistance in resource-limited settings.

Detection of Non-Tuberculosis Mycobacteria by Real Time PCR

Background:: Identification of non-tuberculosis mycobacteria by culture and phenotypic description is commonly used; however, it takes 4 to 6 weeks or even a longer time for slow growing species as well as for identification of some species that may be missed by biochemical characteristics methods. This study aimed to evaluate Real Time PCR for Detection of NTM by Amplification of Internal Transcribed Spacer (ITS) and 16S rRNA. Methods:: In our investigation, using Real Time PCR and two pairs of unique primers targeted to ITS and 16S rRNA genes as well as Beta-actin as an internal control, Non tuberculosis mycobacteria species were detected. Results:: Real time PCR was performed on the prepared dilutions. In addition, the threshold of sensitivity in this study was 10pg. To test the specificity, the genome of several bacteria responsible for respiratory infections was used, in which only the test response related to the non-tuberculosis mycobacterium genome and internal control was positive. Conclusion:: In this research, an effective and up-to-date Real Time PCR method was used to design a diagnostic kit from all aspects. To avoid any error or mistake and to minimize the false results, internal control was used. The ability to design diagnostic kits allows us to increase efficiency, minimize mistakes, and save a considerable amount of time and cost.

Clinical characteristics and drug resistance profile of 276 non-tuberculosis mycobacterium pulmonary disease patients

Objective To investigate and analyze the clinical characteristics, pathogen species composition and drug sensitivity to first-line anti-tuberculosis drugs of patients with non-tuberculosis mycobacterium (NTM) pulmonary disease from Nanyang, Henan province, and provide evidence to improve the diagnosis and treatment of NTM pulmonary disease. Methods A total of 276 patients with NTM pulmonary disease (all living in Nanyang) who visited the Sixth People's Hospital of Nanyang City from January 2015 to December 2019 were enrolled retrospectively. The baseline data, clinical symptoms, imaging manifestations and other information of the patients were collected, PCR reverse dot blot hybridization was used to identify NTM strains, and absolute concentration method was used for drug sensitivity test. SPSS18.0 was used for statistical analysis. Results From 2015 to 2019, the identification rates of NTM patients in suspected tuberculosis (TB) patients were 1.05%, 1.11%, 1.23%, 1.54% and 1.70%, respectively. Among the 276 NTM patients, men accounted for 61.59% (170/276), and the average age of the patients at onset was（51.21 ± 8.33）years. The main clinical manifestations were cough, expectoration, chest tightness, shortness of breath or anhelation. The most common imaging manifestations were pleural thickening (53.62%, 148/276), bronchiectasis (42.39%, 117/276) and cavity (40.58%, 112/276). Eight NTM strains were identified, and the top four strains were Mycobacterium intracellular (32.61%, 90/276), Mycobacterium kansaii (23.91%, 66/276), Mycobacterium abscessus (21.38%, 59/276) and Mycobacterium avium (14.49%, 40/276). The drug resistance rates of the four major NTM to isoniazid, rifampicin, ethambutol, streptomycin, kanamycin and ofloxacin were 92.55% (236/255), 83.53% (213/255), 54.51% (139/255), 89.02% (227/255), 78.82% (201/255) and 64.31% (164/255), respectively. In NTM subspecies, M. aeruginosa was more likely to cause symptoms such as fever, chest tightness, shortness of breath or anhelation and imaging findings such as bronchiectasis and cavity compared with M. avium. Conclusion The identification rate of NTM pulmonary disease in Nanyang increased year by year. The clinical characteristics were similar to those of patients with pulmonary TB. Close attention should be paid to the patients with risk factors and complications in clinical practice, and the imaging manifestations and bacteriological characteristic should be differentiated. The common NTM strains had high resistance to the first-line anti-TB drugs, so the treatment plan should be standardized.

Microflora of autopsy material from HIV-infected patients

According to literature, lungs are affected in 25 – 60% of HIV-infected individuals. The most common opportunistic disease in this group of patients is pneumonia of various etiology. HIV-positive patients show a wide microbiological landscape of pneumonia pathogens.The aim of the study was to determine the structure of the microflora of autopsy material obtained from the lungs of HIV-infected patients.Methods. 83 samples of autopsy material from patients with HIV infection and 27 samples from HIV-negative patients were analyzed. The deceased were treated in the pulmonary departments of Samara with a diagnosis of community-acquired pneumonia. Due to the large variety of microorganisms recovered from the autopsy material, the microorganisms were divided into several groups: Enterobacteria, non-fermentative gram-negative bacteria, Enterococci, Staphylococci, Sterptococci, and Corynebacteria. The fungi were divided into two subgroups. The first included Candida spp., the second included other types of fungi. The tests for pneumocystis infection, Mycobacterium tuberculosis, and non-tuberculosis mycobacteria were not performed.Results. Enterobacteria were found in 39.2%, non-fermentative gram-negative bacteria – in 27.3% and Enterococci – in 19.6% of the HIV-infected patients. Staphylococcus spp. were found in 4.2%, and Corynebacteria and Streptococci in 0.7% of those patients. Candida spp. accounted for 6.3%, other fungi were found in 2.0% of cases. Enterobacteria and non-fermentative gram-negative bacteria were found in 26.2% of HIV-negative patients. Enterococcus spp. – 21.4%, Staphylococci – 7.1%, Streptococci – 2.4% of those patients. Candida spp. 16.7% were found in 16.7% of HIV-negative patients. Enterobacteria were found significantly more often in the autopsy material from HIV-infected patients. The study shows the high importance of gram-negative flora as a causative agent of infectious lesions of the lung tissue. At the same time, we statistically confirmed an increase in prevalence of Enterobacteria in HIV-positive patients. Although these pathogens are not always detected in the sputum from HIV-infected patients with bronchopulmonary disorders, they are detected only by autopsy examination of the lung tissue.Conclusion. Diagnosis and treatment of pneumonia in HIV-infected patients with severe immunodeficiency have a few specific features, such as increasing the frequency of sputum culture, reducing the number of errors at the preanalytical stage of collecting the biomaterial, ensuring proper conditions for its transportation to a bacteriological laboratory, improving the quality of analysis. The physicians should be aware of the possible detection of gram-negative flora as a causative agent of lung infections and the possible change of the causative agent.

Nanoparticulate β-Cyclodextrin with Gallium Tetraphenylporphyrin Demonstrates in Vitro and in Vivo Antimicrobial Efficacy against Mycobacteroides abscessus and Mycobacterium avium.

The emergence of drug-resistant pathogens causes the greatest challenge for drug development research. Recently, gallium(III)-based compounds have received great attention as novel antimicrobial agents against drug-resistant pathogens. Here, we synthesized a new β-cyclodextrin Ga nanoparticle (CDGaTP) using Ga tetraphenylporphyrin (GaTP, a hemin analogue) and β-cyclodextrin. The newly synthesized nanoparticle was nontoxic and efficient at a single dose, showing sustained drug release for 15 days in vitro. CDGaTP's activity with transferrin or lactoferrin was tested, and synergism in activity was observed against nontuberculosis mycobacteria (NTM), Mycobacterium avium (M.avium), and Mycobacteroides abscessus. Human serum albumin (HSA) decreased the efficacy of both GaTP and CDGaTP in a concentration-dependent manner. The NTMs incubated with GaTP or CDGaTP significantly produced reactive oxygen species (ROS), indicating potential inhibition of antioxidant enzymes, such as catalase. The single-dose CDGaTP displayed a prolonged intracellular inhibitory activity in an in vitro macrophage infection model against both NTMs. In addition, CDGaTP, not GaTP, was effective in a murine lung M.avium infection model when delivered via intranasal administration. These results suggest that CDGaTP provides new opportunities for the development of gallium-porphyrin based antibiotics.

Some Opportunistic Infections in Childhood: Causes, Diagnosis and Treatment Methods

The article discusses the features of the emergence and development of the so-called opportunistic infections that occur in children with weakened immunity.
 Such infections are often caused by non-tuberculosis mycobacteria, fungi, and herpesviruses, and such infections can also develop in children who are treated with immunobiological drugs. Since these are serious and rapidly developing diseases, the diagnosis should be made at the earliest stages of the development of the disease and be based on the features of clinical phenomena described in the literature and anamnesis data indicating the presence of such infections. The earlier an infection is detected, the faster it will be possible to start appropriate therapy.
 The relevance of this topic is due to the fact that, despite the improvement in the diagnosis of opportunistic infections in recent years, they still are a problem for pediatricians, who are not always able to diagnose them in time. Accordingly, the pediatrician should, in case of suspicion of the presence of such an infection in a child patient, contact specialists who have experience in their treatment in order to take timely measures to stop the development of the disease.
 The aim of the study is to consider the features of some opportunistic infections in childhood, their causes, diagnosis, and treatment methods.

Non-tuberculous mycobacteria in phthisiopulmonary practice in the Republic of Uzbekistan

The objective of the study: monitoring the spectrum of non-tuberculous mycobacteria isolated from patients who referred for medical care to the Republican Specialized Scientific and Practical Medical Center of Phthisiology and Pulmonology.Subjects and methods. The diagnostic procedure of ATS/IDSA was used to define if the patient suffers from mycobacteriosis. The following specimens were collected to isolate non-tuberculosis mycobacteria: sputum, bronchoalveolar lavage fluid, feces, pleural fluid, surgical and biopsy specimens, and urine. The following tests were performed: Ziehl-Nielsen microscopy, microscopy stained by auramine-O, cultures by Middlebrook 7H9 in BACTEC™ MGIT™ 960 System, Becton Dickinson, USA. Non-tuberculosis mycobateria were differentiated from mycobacterium tuberculosis complex using the sdmpt64 chromatographic test (SD Bioline TBAg MPT64 test, Korea). The non-tuberculosis species were defined by the hybridization technology of DNA* strips GenoType Mycobacterium AS/CM, version 1.0.Results. Of 14,544 patients with suspected respiratory tuberculosis, non-tuberculous mycobacteria were detected in 38 (0.26%) of them, 17 (44.7%) patients had Mycobacterium avium complex, in them there were 26 men (68.4%) and 12 (31.6%) women. Non-tuberculosis mycobacteria were isolated mainly from sputum – in 27 (71.2%) patients and urine – in 6 (15.7%) patients. In 26 (68.4%) patients, mycobacteriosis was caused by slow-growing non-tuberculosis mycobacteria, of which Mycobacterium avium complex prevailed – in 17 people as well as Mgordonae – in 8 patients. Rapidly growing non-tuberculosis mycobacteria were identified in 12 (31.6%) patients, they included M. fortuitum (5 cases) and M. chelonae (4) prevailed.

Systemic infections associated with tattoos or permanent makeup: A systematic review

IntroductionThe aim of this article is to summarize published information on systemic infective complications of tattoos to gain an update of the current picture. MethodsA literature search was performed in PubMed database (2009–2019), and compared with a search without year restriction. Eligibility criteria were studies on systemic tattoo-related infections, including case reports, case series, outbreak investigations, reviews, and systematic reviews. ResultsWe identified 17 manuscripts with systemic infections between 2009 and 2019, with one reported fatality. In contrast to the historical records, no reports of systemic tuberculosis, syphilis or viral (hepatitis or HIV) infections were reported within the study period. A few sporadic cases or Mycobacterium leprae (India) or regional lymphadenopathy associated with skin lesions in non-tuberculosis mycobacteria were identified. Persistent fever with rigour was common in bacterial bloodstream infections. One episode of staphylococcal toxic shock syndrome and several episodes of septic shock were reported, associated with cellulitis or necrotizing fasciitis within two weeks of the procedure, predominantly caused by pyogenic bacteria (S. aureus or streptococcus). Identification of lung or systemic embolisms in the absence of local symptoms, was indicative of (right or left) infective endocarditis. ConclusionsBacterial bloodstream infections should be considered in subjects developing fever and rigour after tattoos, regardless of local symptoms. A shift in causative organisms has been documented, when comparing with historical reports. NTM are emerging organisms causing lymphadenopathy. Strict hygiene conditions are essential when performing a tattoo.

Mycobacteria Tales Post Lung Transplantation: An Indian Experience

Purpose Post-lung transplant infection is one of the leading causes of morbidity and mortality. The diagnosis of pulmonary tuberculosis can be even more challenging in the setting of lung transplantation where other pulmonary complications can make diagnosis difficult. Infection may be caused by reactivation of a primary infection in the recipient, reactivation of a lesion from the donor lung, or primary infection. We present a retrospective study from India to define the incidence, clinical presentation, and course of Mycobacterial infections in a cohort of lung transplant recipients. Methods Between 1 April 2017 and 1 September 2020, 140 lung transplants were performed. Serial bronchoscopies with transbronchial biopsies and bronchioalveolar lavages were performed during follow-up. Explant Histopathological findings were taken into consideration. Pleural based interventions in the form of Tapping or CT guided Pleural biopsy were noted. Similarly, NTM infections were documented with species identification on cultures. Results 12 (8.57%) Recipients had Mycobacterial Infections post lung transplant. Mycobacterium Tuberculosis species was diagnosed in 9 patients (75%). Rest 3 recipients (25%) had Non-Tuberculosis Mycobacterium species (2 Mycobacterium abscessus and 1 Mycobacterium fortuitum). In 4 MTB patients, diagnosis was determined from the explanted lungs histopathology. The remainder were diagnosed during postoperative period. Median time to diagnosis was 40.5 days . Radiographic findings (HRCT scan) were Consolidations (n=3), Pleural Effusion (n=3), Tree in Bud (n=1), Cavity (n=1), no typical radiographical presentations (n=4) All patients responded well to anti-mycobacterial therapy. 3 recipients with NTM received individualised treatment for the same. Among these 12 recipients, 3 recipients expired with death attributable to Mycobacterial disease in only 1 recipient. Conclusion Mycobacterial Infection is not rare in lung transplant recipients and can be managed successfully with individualised antimycobacterial therapy. Non-Rifampicin based AntiMycobacterial treatment strategy is preferred to avoid drug interactions with immunosuppressants. Explant Lung Histopathology is an important tool to diagnose Tuberculosis specifically in Indian settings. NTM infection is not uncommon post lung transplantation & should have low threshold for diagnosis.

Potential zoonotic pathogens hosted by endangered bonobos

Few publications, often limited to one specific pathogen, have studied bonobos (Pan paniscus), our closest living relatives, as possible reservoirs of certain human infectious agents. Here, 91 stool samples from semicaptive bonobos and bonobos reintroduced in the wild, in the Democratic Republic of the Congo, were screened for different infectious agents: viruses, bacteria and parasites. We showed the presence of potentially zoonotic viral, bacterial or parasitic agents in stool samples, sometimes coinfecting the same individuals. A high prevalence of Human mastadenoviruses (HAdV-C, HAdV-B, HAdV-E) was observed. Encephalomyocarditis viruses were identified in semicaptive bonobos, although identified genotypes were different from those identified in the previous fatal myocarditis epidemic at the same site in 2009. Non-pallidum Treponema spp. including symbiotic T. succinifaciens, T. berlinense and several potential new species with unknown pathogenicity were identified. We detected DNA of non-tuberculosis Mycobacterium spp., Acinetobacter spp., Salmonella spp. as well as pathogenic Leptospira interrogans. Zoonotic parasites such as Taenia solium and Strongyloides stercoralis were predominantly present in wild bonobos, while Giardia lamblia was found only in bonobos in contact with humans, suggesting a possible exchange. One third of bonobos carried Oesophagostomum spp., particularly zoonotic O. stephanostomum and O. bifurcum-like species, as well as other uncharacterized Nematoda. Trypanosoma theileri has been identified in semicaptive bonobos. Pathogens typically known to be transmitted sexually were not identified. We present here the results of a reasonably-sized screening study detecting DNA/RNA sequence evidence of potentially pathogenic viruses and microorganisms in bonobo based on a noninvasive sampling method (feces) and focused PCR diagnostics.

PCR-reverse blot hybridization assay in respiratory specimens for rapid detection and differentiation of mycobacteria in HIV-negative population

BackgroundRapid identification of pathogenic Mycobacterium species is critical for a successful treatment. However, traditional method is time-consuming and cannot discriminate isolated non-tuberculosis mycobacteria (NTM) at species level. In the retrospective study, we evaluated the clinical applicability of PCR-reverse blot hybridization assay (PCR-REBA Myco-ID) with clinical specimens for rapid detection and differentiation of mycobacterial species.MethodsA total of 334 sputum and 362 bronchial alveolar lavage fluids (BALF) from 696 patients with mycobacterium pulmonary disease (MPD) and 210 patients with non-mycobacterium pulmonary disease used as controls were analyzed. Sputum or BALF were obtained for MGIT 960-TBc ID test and PCR-REBA Myco-ID assay. High resolution melt analysis (HRM) was used to resolve inconsistent results of MGIT 960-TBc ID test and PCR-REBA Myco-ID assay.ResultsA total of 334 sputum and 362 BALF specimens from 696 MPD patients (292 MTB and 404 NTM) were eventually analyzed. In total, 292 MTBC and 436 NTM isolates (mixed infection of two species in 32 specimens) across 10 Mycobacterium species were identified. The most frequently isolated NTM species were M. intracellulare (n = 236, 54.1%), followed by M. abscessus (n = 106, 24.3%), M. kansasii (n = 46, 10.6%), M. avium (n = 36, 8.3%). Twenty-two cases had M. intracellulare and M. abscessus mixed infection and ten cases had M. avium and M. abscessus mixed infection. A high level of agreement (n = 696; 94.5%) was found between MGIT 960-TBc ID and PCR-REBA Myco-ID (k = 0.845, P = 0.000). PCR-REBA Myco-ID assay had higher AUC for both MTBC and NTM than MGIT 960-TBc ID test.ConclusionPCR-REBA Myco-ID has the advantages of rapid, comparatively easy to perform, relatively low cost and superior accuracy in mycobacterial species identification compared with MGIT 960-TBc ID. We recommend it into workflow of mycobacterial laboratories especially in source-limited countries.

Genetic Manipulation of Non-tuberculosis Mycobacteria.

Non-tuberculosis mycobacteria (NTMs) comprise a large group of organisms that are phenotypically diverse. Analysis of the growing number of completed NTM genomes has revealed both significant intra-genus genetic diversity, and a high percentage of predicted genes that appear to be unique to this group. Most NTMs have not been studied, however, the rise in NTM infections in several countries has prompted increasing interest in these organisms. Mycobacterial research has recently benefitted from the development of new genetic tools and a growing number of studies describing the genetic manipulation of NTMs have now been reported. In this review, we discuss the use of both site-specific and random mutagenesis tools in NTMs, highlighting the challenges that exist in applying these techniques to this diverse group of organisms.

Pediatric Spinal Subdural Abscesses: A Report of Three Consecutive Patients

Background: Despite technological advances in medical treatment, the prognosis of the rarely reported spinal subdural abscesses (SSAs) has remained a serious entity largely unaffected, especially when they are diagnosed late. In this study, the authors aimed to present the surgical outcomes of 3 consecutive pediatric patients with SSA. Materials and Methods: We retrospectively reviewed the medical records of pediatric patients with spinal lesions who underwent surgery at 2 neurosurgical centers spanning 7 years, from 2012 to 2019. All pediatric patients who were diagnosed with SSA (n = 3) were selected as the core sample for this study. Results: Three pediatric patients (2 females and 1 male) with SSA were surgically treated. Holocord SSA was observed in 1 patient. The mean age was 7.1 ± 7.7 years. The most common presenting symptoms were gait disturbance and weakness of lower extremities (100%). The mean preoperative course was 5.7 ± 4.0 weeks. The causative pathogens were Escherichia coli (E. coli) and M. tuberculosis. In the 2nd case, the pathogen was non-tuberculosis mycobacterium in the extramedullary abscess. In the 44th postoperative month, she underwent surgery for intramedullary abscess. The causative pathogen was E. coli. Except for 1 male adolescent who presented with severe clinical status (paraplegic), the improvement was observed in all patients at their last follow-up after 50.3 ± 43.5 months of average. Conclusions: Drainage followed by appropriate antibiotics is the optimal treatment for SSAs. Early diagnosis and urgent surgical treatment are essential for a good prognosis. All surgically treated SSA patients with neurological deficits were rehabilitated with physical therapy postoperatively.

Microbiological pattern of prosthetic hip and knee infections: a high-volume, single-centre experience in China.

Introduction. Prosthetic joint infection (PJI) is a serious complication after arthroplasty, which results in high morbidity, prolonged treatment and considerable healthcare expenses in the absence of accurate diagnosis. In China, microbiological data on PJIs are still scarce.Hypothesis/Gap Statement. The incidence of PJI is increasing year by year, and the proportion of drug-resistant bacteria infection is nicreasing, which brings severe challenges to the treatment of infection.Aim. This study aimed to identify the pathogens in PJIs, multi-drug resistance, and evaluate the effect of the treatment regimen in patients with PJI.Methodology. A total of 366 consecutive cases of PJI in the hip or knee joint were admitted at the Orthopedic Surgery Center in Zhengzhou, China from January 2012 to December 2018. Infections were confirmed in accordance with the Infectious Diseases Society of America and the Musculoskeletal Infection Society (MSIS) criteria. Concurrently, patient demographic data, incidence and antibiotic resistance were investigated. Statistical differences were analysed using Fisher's exact test or chi-square test.Results. Altogether, 318 PJI cases satisfying the inclusion criteria were enrolled in this study, including 148 with hip PJIs and 170 with knee PJIs. The average age of patients with hip PJIs was lesser than that of patients with knee PJIs (56.4 vs. 68.6 years). Meanwhile, coagulase-negative staphylococcus (CNS, n=81, 25.5 %) was the predominant causative pathogen, followed by Staphylococcus aureus (n=67, 21.1 %). Methicillin-resistant Staphylococcus (MRS) was identified in 28.9 % of PJI patients. In addition, fungus accounted for 4.8 % (n=15), non-tuberculosis mycobacterium accounted for 1.6 % (n=5), polymicrobial pathogens accounted for 21.7 % (n=69), and Gram-negative bacteria accounted for 7.9 % (n=25) of the total infections. The results of antibiotic susceptibility testing showed that gentamicin and clindamycin β-lactam antibiotics were poorly susceptible to Gram-positive isolates, but they were sensitive to rifampicin, linezolid and vancomycin. While antibiotics such as amikacin and imipenem were effective against Gram-negative bacteria, there was a high resistance rate of other pathogens to gentamicin, clindamycin and some quinolone antibacterial drugs. Empirical antibiotic treatment should combine vancomycin and cephalosporin, levofloxacin or clindamycin. When the pathogen is confirmed, the treatment should be individualized.Conclusions. The prevalence of culture-negative PJIs is still very high. Gram-positive bacteria are still the main type of pathogens that cause PJIs. Attention should be paid to the high incidence of MRS, such as MRSA and MR-CNS, among PJI patients. Empirical antibiotic treatment should cover Gram-positive isolates, especially Staphylococcus.

Same Game, Different Players: Emerging Pathogens of the CF Lung.

The survival rates of individuals with cystic fibrosis (CF) have significantly increased as a result of improved therapies, such as the inclusion of cystic fibrosis transmembrane conductance regulator (CFTR) modulators for some mutations. However, microbial infection of the airways remains a significant clinical problem. The well-known pathogens Pseudomonas aeruginosa and Staphylococcus aureus continue to establish difficult-to-treat infections in the CF lung. However, in recent years, there has been an increased prevalence of both Aspergillus fumigatus (Af) and non-tuberculous mycobacteria (NTM) species isolated from CF patient sputa. The emergence of these pathogens opens an important area of discussion about multikingdom infections, specifically, how interspecies interactions have the potential to shape the course of infection, such as tolerance to host immune defenses and antimicrobial therapies. Their ability to establish themselves in an existing polymicrobial environment suggests to us that microbial interactions play a significant role, and characterizing these mechanisms and understanding their implications will be critical to the future development of better antimicrobial therapies. With this minireview, we hope to inspire conversations about and demonstrate the merit of more research in this area.IMPORTANCE Incidences of non-tuberculous mycobacteria (NTM) and Aspergillus fumigatus have increased around the world over the past decade and have become a significant health threat to immunocompromised individuals such as those with cystic fibrosis (CF). CF is characterized by the buildup of mucus in the lungs which become chronically infected by a myriad of pathogens. The emergence of these pathogens in established infection sites raises many questions about the microbial ecosystems they become a part of, specifically how changes in these ecosystems impact disease outcomes. Understanding how microbial communities establish and maintain themselves despite medical treatment and host immune defenses will be critical for the development of improved therapeutic strategies.

A novel DNA chromatography method to discriminate Mycobacterium abscessus subspecies and macrolide susceptibility.

BackgroundThe clinical impact of infection with Mycobacterium (M.) abscessus complex (MABC), a group of emerging non-tuberculosis mycobacteria (NTM), is increasing. M. abscessus subsp. abscessus/bolletii frequently shows natural resistance to macrolide antibiotics, whereas M. abscessus subsp. massiliense is generally susceptible. Therefore, rapid and accurate discrimination of macrolide-susceptible MABC subgroups is required for effective clinical decisions about macrolide treatments for MABC infection. We aimed to develop a simple and rapid diagnostic that can identify MABC isolates showing macrolide susceptibility.MethodsWhole genome sequencing (WGS) was performed for 148 clinical or environmental MABC isolates from Japan to identify genetic markers that can discriminate three MABC subspecies and the macrolide-susceptible erm(41) T28C sequevar. Using the identified genetic markers, we established PCR based- or DNA chromatography-based assays. Validation testing was performed using MABC isolates from Taiwan.FindingWe identified unique sequence regions that could be used to differentiate the three subspecies. Our WGS-based phylogenetic analysis indicated that M. abscessus carrying the macrolide-susceptible erm(41) T28C sequevar were tightly clustered, and identified 11 genes that were significantly associated with the lineage for use as genetic markers. To detect these genetic markers and the erm(41) locus, we developed a DNA chromatography method that identified three subspecies, the erm(41) T28C sequevar and intact erm(41) for MABC in a single assay within one hour. The agreement rate between the DNA chromatography-based and WGS-based identification was 99·7%.InterpretationWe developed a novel, rapid and simple DNA chromatography method for identification of MABC macrolide susceptibility with high accuracy.FundingAMED, JSPS KAKENHI

Comparison of Different Diagnostic Techniques for the Identification of Pulmonary Tuberculosis and Using Serum Tnf-? Levels as a Biomarker for Evaluating the Severity of Tuberculosis

Background and Objective: Pulmonary tuberculosis(PTB) is a serious disease that mainly affects thelungs. The aim of this study was a Comparison between conventional, and molecular methods to diagnoseM.tuberculosis complex (MTBC) and using serum TNF-? levels as a biomarker for evaluating the severity ofthe disease. Methodology: The study analyzed 586 sputa collected from suspected pulmonary tuberculosispatients (217 female and 369 male) their ages ranged from 16 to 66 years for mycobacteriological study,while the immuno-study included 85 blood samples that collected from eighty patients and five healthypersons (45 male,40 female) their ages (18-66) years. We tested all sputum samples by AFB direct stain andGX methods, while we cultured sputum restrictively for Patients who have two different outcomes by AFBdirect stain and GX methods. TNF-? levels have been measured by sandwich ELISA in patients and healthypersons. Results: Our result showed detection of (135\586) patients infected with M.tuberculosis complex(MTBC) and (1\586) patient infected with non-mycobacterium tuberculosis(NMT) by AFB direct stain andGX methods comparing with culture method as a gold standard. Sensitivity and specificity for AFB directstain and GX techniques for diagnosis PTB were (15.0% and 100% ) and (95.0% and 66.7% ) respectively.On the other side, TNF-? levels were elevated in inpatient groups compared with healthy persons. The meanof TNF-? levels measured in multidrug resistance tuberculosis (MDR-TB) patients was higher than in othercases. Conclusion: Our findings demonstrate that the sensitivity of the GX technique is higher comparedto other techniques, and serum TNF-? levels can be used as a biomarker for evaluating the severity ofpulmonary tuberculosis.