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Introduction: Non-tuberculous Mycobacteria (NTM) are mainly environmental but can cause opportunistic infections and diseases in humans and animals. Livestock and wild animals can be infected with NTM. In Argentina, there are native wild species facing conservation risks, and they are the focus of protection and reintroduction projects designed to preserve biodiversity in various ecoregions. The aim of this study was to report the presence of NTM in samples collected from four endangered native wild species from nine Argentine provinces, as part of their pre-release health assessment. Methods: A total of 165 samples from giant anteater, peccary, tapir and pampas deer were obtained, these included either bronchoalveolar or endotracheal lavages, or oropharyngeal, nasopharyngeal or tracheal swabs. Bacteriological culture followed by molecular identification and sequencing were performed. Results: A total of 27 NTM were detected, including Mycobacterium avium subsp. hominissuis, M. intracellulare, M. terrae, M. gordonense, M. kumamotonense, M. fortuitum, M. saskatchewanense, and M. genavense. Results revealed a 16,36% NTM recovery rate, with the giant anteater showing the highest prevalence among the mammals under study. Discussion: In Argentina, due to extensive production systems, the interaction between domestic and wild species sharing the same environment is frequent, increasing the exposure of all the species to these NTM. In this way, the transmission of infectious agents from one to another is feasible. Moreover, NTMs might interfere with the diagnosis of bovine tuberculosis and paratuberculosis. These findings emphasize the importance of active health surveillance in conservation programs. It highlights the need to address NTM epidemiology in wildlife and its impact on conservation and public health.

RESUMEN

The results of evaluating the effectiveness of the use of liquid transport media at the preanalytical stage of bacteriological diagnosis of diphtheria infection are presented. A typical toxigenic strain of C. diphtheriae biovar gravis № 665 was used. The experiments were carried out using a laboratory-prepared medium based on GRM-broth (State research center for applied biotechnology and microbiology, Obolensk), a transport system with a fleecy probe swab (DELTALAB) and a transport system ∑-Transwab ® with a polyurethane Sigma-swab (Medical Wire & Equipment Co. (Bath) Ltd.). The tampons were pooled with a 24-hour bacterial culture of C. diphtheriae, then immediately seeded on Tellurite-containing blood agar. Storage conditions were simulated for 6-24 hours: at room conditions +(20-25)° C, in the refrigerator +(4-8)° C, in a thermostat +(37±1)° C. Storage of C. diphtheriae was most optimal on two liquid transport systems in a refrigerator +(4-8)° C for 6 and 24 hours; in room conditions +(20-25)° C - there was a decrease in seeding after 6 hours and loss of pathological material after 24 hours, more pronounced on a fleecy probe swab; under thermostat conditions +(37±1)° C on both transport systems, a decrease in seeding was noted after 6 hours and a complete loss of pathological material after 24 hours. The results obtained demonstrated the efficiency of using the Amies liquid transport medium and justify the need to develop a domestic analogue of the transport system based on the Amies liquid medium for the bacteriological diagnosis of diphtheria infection.

Asunto(s)

Corynebacterium diphtheriae , Difteria , Corynebacterium , Medios de Cultivo , Difteria/diagnóstico , Humanos , Manejo de Especímenes

RESUMEN

The results of evaluating the effectiveness of C. diphtheriae inoculation using different types of dry swabs in studies simulating various conditions of its storage at the preanalytical stage of a laboratory study for diphtheria are presented. A typical toxigenic strain of C. diphtheriae biovar gravis No. 665 was used. A commercial dry, sterile cotton swab probe (Ningbo Greetmed Medical Instruments Co., LTD, China), a commercial dry, sterile swab probe (plastic and viscose) (COPAN, Italy), tufters with a fluffy probe-tampon on a polystyrene applicator, standard (DELTALAB, SL, Spain). The tampons were pooled with a 24-hour bacterial culture of C. diphtheriae, then immediately seeded on Tellurite-containing blood agar and Corynebacagar. Storage conditions were simulated for 3 hours: at room conditions +(20-25)°C, in the refrigerator +(4-8)°C, in a thermostat +(37±1)°C. Optimal storage of C. diphtheriae on all three types of dry swabs at + (4-8) ° C; at +(20-25)° C - growth is observed when seeding from a cotton swab; in a swab with a fleecy probe-tampon, a decrease in the inoculation of C. diphtheriae was noted; when using a viscose swab - a significant loss of C. diphtheriae. At +(37±1)°C, a significant decrease in the inoculation of C. diphtheriae on all three types of tampons was noted, up to the absence of growth when using a viscose tampon. To exclude the loss of C. diphtheriae, it is necessary to observe the conditions for taking and storing biological material at the preanalytical stage of a laboratory study, which will improve the quality of laboratory microbiological studies for diphtheria infection.

Asunto(s)

Corynebacterium diphtheriae , Difteria , Corynebacterium , Medios de Cultivo , Difteria/diagnóstico , Humanos

RESUMEN

The results of comparative experimental studies of identification of nontoxigenic C. diphtheriae strain by three different commercial laboratories are presented. A typical nontoxigenic strain of C. diphtheriae biovar mitis was used. For the studies, three lines of ten-fold dilutions of bacterial culture were prepared, followed by control planting on the medium and counting CFU/ml. In the experiment, tampons were pooled with a 24-hour bacterial culture of a nontoxigenic C. diphtheriae strain. Tampons were provided from three different laboratories - ∑-Transwab® with Ames liquid medium (from the first and second laboratories) and a viscose tampon with coal medium (from the third laboratory). After pooled, tampons were delivered to commercial laboratories. And as a result of the experiment, Corynebacterium spp. was identified in first laboratory (103 CFU/tamp), S. epidermidis (102 CFU/ml) - in second laboratory and nontoxigenic C. diphtheriae biovar gravis - in third laboratory. The study indicates that there is a need to the supervision of bacteriological investigations conducted in various laboratories. This will improve the quality of investigations on diphtheria infection and identify of diphtheria carrier, which is a reservoir of the causative agent of diphtheria, and will contribute to the maintenance of sanitary and epidemiological well-being in our country.

Asunto(s)

Corynebacterium diphtheriae , Difteria , Corynebacterium diphtheriae/genética , Medios de Cultivo , Difteria/diagnóstico , Humanos

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The purpose of the work is to evaluate the cultural and morphological properties of colonies of clinically significant corynebacteria on culture mediums for the isolation of corynebacteria. The study used 9 culture mediums for the isolation of corynebacteria: a culture medium for the isolation of corynebacteria (Corynebacagar); Tellurite-containing blood agars on base - Culture medium № 1 GRM, Culture agar for the cultivation of microorganisms (GRM agar), Culture medium for determining the sensitivity of microorganisms to antibacterial preparations - AGV, culture agar for the cultivation of dry microorganisms (SPA), Clauberg medium II, Hoyle Medium agar (Oxoid), Blood agar base (Conda), Columbia Agar Base (Conda). The work used 7 test strains of microorganisms from the State collections of pathogenic microorganisms - C. diphtheriae biovars gravis, mitis, intermedius, belfanti and subspecies lausannense, C. ulcerans and C.pseudotuberculosis. Studies were carried out in accordance with MUK 4.2.3065-13 «Laboratory diagnosis of diphtheria infection¼. We describe culture-morphological properties of strains on all tested culture mediums the isolation of corynebacteria after 24 and 48 hours of incubation. Analysis of the results on the growth properties of culture mediums showed that all culture mediums had high sensitivity - from dilution 10-7 for all test strains. Colonies of corynebacteria were visually detected on culture mediums after 19-20 hours of cultivation. When cultivating a suspension of corynebacteria from breeding 10-6 on culture mediums, the number of colonies ranged from 95±5 to 120±10. Conclusion. All culture mediums had differential diagnostic properties that ensure the growth of corynebacteria after the day of incubation.

Asunto(s)

Corynebacterium diphtheriae , Difteria , Corynebacterium , Medios de Cultivo , Difteria/diagnóstico , Humanos , Laboratorios

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The purpose of the work was to assess the state of bacteriological diagnosis of diphtheria infection in Russia in order to establish possible reasons for the decrease in the release of C. diphtheriae. The Reference Center for Monitoring the Pathogens of Measles, Rubella, Mumps, Pertussis and Diphtheria in 2018 in 85 subjects of Russia conducted a questionnaire of laboratories of medical organizations and the Centers for Hygiene and Epidemiology of Rospotrebnadzor, carrying out bacteriological studies for diphtheria infection. It was found that the number of studies conducted over the five-year period decreased by 1.2 times. The tendency to decrease the number of bacteriological studies for diphtheria is observed in the territories of almost all federal districts. In 99% and 29% of cases, the institutions of the FBUZ Centers for Hygiene and Epidemiology and medical organizations (MO) and use in their work documents regulating bacteriological studies for diphtheria infection. In a number of territories, the list of documents used includes documents that are invalid or do not define such studies. Most organizations use dry tampons when examining for diphtheria, however, 13.1% and 53.4% of FBUZ Centers for Hygiene and Epidemiology and medical organizations (respectively) use commercial transport environments, which does not comply with regulatory documentation. Analysis of the quality of work of bacteriological laboratories showed shortcomings at the stage of preparation of media (use of donor blood, or absence of addition of blood and potassium tellurite), Elek tests (addition of horse serum or absence of serum to the medium), setting of incomplete biochemical series (absence of tests for urease and nitrate reductase), absence of standard control strains, incomplete volume of internal laboratory quality control. Given the continuing circulation of the pathogen in various countries of the world and in our country, as well as the possibility of imported cases of infection from endemic regions, the analysis was aimed at drawing the attention of specialists to the problem of improving the quality of laboratory diagnosis of diphtheria in Russia.

Asunto(s)

Corynebacterium diphtheriae , Difteria , Técnicas de Laboratorio Clínico , Medios de Cultivo , Difteria/diagnóstico , Difteria/epidemiología , Humanos , Federación de Rusia/epidemiología

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Tuberculosis (TB) remains a leading killer among infectious diseases of humans worldwide. Delayed diagnosis is a crucial problem in global TB control programs. Bacteriological methods currently used to diagnose TB in endemic countries take up to 8 weeks, which poses a significant delay in starting antibiotic therapy. The presence of a heterogeneous population of Mycobacterium tuberculosis, the causative agent of TB, is among the reasons for delayed diagnosis by bacteriological methods. Previously, it has been shown that mycobacterial resuscitation-promoting factors (RPFs), a family of proteins secreted by actively growing bacteria into the media, are capable of activating the growth of dormant bacteria, thus enhancing the detection of bacilli in the sputum of confirmed TB cases. However, the variability in bacterial resuscitation by RPF in the sputum of suspected pulmonary TB cases that showed differential smear and/or culture positivity during diagnosis has not been fully explored. Here, we report the presence of non-replicating bacteria in the sputum of suspected TB cases that show differential growth response to RPF treatment. Using crude and recombinant RPF treatment, we show improved sensitivity and reduced time to detect bacilli in the sputum samples of smear-positive/culture-negative or smear-negative/culture-negative cases. We also report the phenotypic heterogeneity in the RPF responsiveness among Mtb strains using an in vitro dormancy model. Our findings have implications for improving the bacteriological diagnostic modalities currently used to diagnose TB in endemic countries.

RESUMEN

This work aims at describing the diversity of osteomyelitis of the jaw (OJ) and at assessing the relevance of a new method designed to avoid salivary contamination during bone sampling in order to improve microbiological analysis and clinical decision-making. We reviewed medical and microbiological data of patients with a suspected OJ based on clinical and/or CT-scan signs and at least one bone sample made for microbiological analysis. During the study period, a new procedure for intraoral bone sampling was elaborated by surgeons and infectious diseases specialists authoring this article (based on stratified samples, cleaning of the surgical site and change of instruments between each sample). A comparison of the microbiological analyses between the two procedures was performed. From 2012 to 2017, 56 patients were included. Median age was 58 years (11-90), sex ratio: 1.24. Main risk factors were having a dental disease (n = 24) or cancer (n = 21). Nineteen patients with the new sample procedure were compared to 37 patients with standard procedure, especially non-cancer patients (n = 16 and 19, respectively). With the new procedure, a median of 3 (1-7) microorganisms per sample was recovered, vs. 7 (1-14) with the former (p < 0.001), a significant decrease of the microbial density was observed for all types of microbes, especially in deeper samples and cultures were more frequently sterile. The way sampling is managed deeply influences microbiological analysis. This strategy facilitates the distinction between pathogens and contaminants and should constitute the first step toward an evidence-based antimicrobial strategy for OJ.

Asunto(s)

Infecciones Bacterianas/diagnóstico , Biopsia/métodos , Huesos/microbiología , Maxilares/microbiología , Osteomielitis/diagnóstico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bacterias/clasificación , Bacterias/aislamiento & purificación , Infecciones Bacterianas/clasificación , Infecciones Bacterianas/microbiología , Biopsia/efectos adversos , Biopsia/instrumentación , Huesos/patología , Niño , Femenino , Humanos , Maxilares/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Neoplasias/complicaciones , Osteomielitis/microbiología , Estudios Retrospectivos , Factores de Riesgo , Saliva/microbiología , Tomografía Computarizada por Rayos X/métodos , Adulto Joven

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BACKGROUND AND AIM: In the last decades, the inhabitants of the Romanian region known as Jiu Valley underwent changes in their social and economic status which determined changes in behaviour and health, which influenced their general health condition. One of the consequences was the exacerbation of tuberculosis. In order to control this situation, there was a need to increase the efficiency of diagnosis. This optimization can be reached by a better detection of mycobacterium infection, optimal isolation of strains and identification of the resistance of strains to antituberculous drugs. METHODS: In order to identify the best diagnostic modality, we compared the efficacy of the classical bacteriological diagnosis, still performed in the field, to the modern methods of molecular biology. The study included two groups, one represented by 213 patients who were investigated using the classical bacteriological methods, and 49 who were diagnosed using the PCR method. RESULTS: The tuberculosis patients who have been evaluated only with the classical bacteriological methods where diagnosed as TB positive and treated according to the national guidelines, which are in agreement with the international guidelines. The PCR diagnostic methods had a superior diagnostic value compared to the traditional bacteriological method. CONCLUSIONS: The results revealed the superiority of the modern molecular biology methods based on PCR. However the bacteriological method remains useful in areas where PCR cannot be afforded.

RESUMEN

INTRODUCTION: Mycobacterium abscessus is an emerging mycobacteria that is responsible for lung diseases and healthcare-associated extrapulmonary infections. Recent findings support its taxonomic status as a single species comprising 3 subspecies designated abscessus, bolletii and massiliense. We performed a review of English-language publications investigating all three of these subspecies. Areas covered: Worldwide, human infections are often attributable to environmental contamination, although the isolation of M. abscessus in this reservoir is very rare. Basic research has demonstrated an association between virulence and cell wall components and cording, and genome analysis has identified gene transfer from other bacteria. The bacteriological diagnosis of M. abscessus is based on innovative tools combining molecular biology and mass spectrometry. Genotypic and phenotypic susceptibility testing are required to predict the success of macrolide (clarithromycin or azithromycin)-based therapeutic regimens. Genotyping methods are helpful to assess relapse and cross-transmission and to search for a common source. Treatment is not standardised, and outcomes are often unsatisfactory. Expert commentary: M. abscessus is still an open field in terms of clinical and bacteriological research. Further knowledge of its ecology and transmission routes, as well as host-pathogen interactions, is required. Because the number of human cases is increasing, it is also necessary to identify more active treatments and perform clinical trials to assess standard effective regimens.

Asunto(s)

Antibacterianos/uso terapéutico , Azitromicina/uso terapéutico , Claritromicina/uso terapéutico , Infecciones por Mycobacterium/epidemiología , Mycobacterium/aislamiento & purificación , Infecciones del Sistema Respiratorio/epidemiología , Antibacterianos/administración & dosificación , Azitromicina/administración & dosificación , Claritromicina/administración & dosificación , Fibrosis Quística/tratamiento farmacológico , Fibrosis Quística/epidemiología , Fibrosis Quística/microbiología , Humanos , Mycobacterium/clasificación , Mycobacterium/efectos de los fármacos , Mycobacterium/genética , Infecciones por Mycobacterium/diagnóstico , Infecciones por Mycobacterium/tratamiento farmacológico , Infecciones por Mycobacterium/microbiología , Prevalencia , Infecciones del Sistema Respiratorio/tratamiento farmacológico , Infecciones del Sistema Respiratorio/microbiología , Resultado del Tratamiento , Virulencia

RESUMEN

Com o objetivo de avaliar o diagnóstico bacteriológico e controle de tratamento da tuberculose, os autores analisaram, nos anos de 1999 e 2000, os exames processados pelo Instituto Adolfo Lutz de Campinas e laboratórios de sua abrangência das Diretorias Regionais de Saúde (DIR)-XII, XV e XX, que atendem 93 municípios. Neste período foram realizados 53.341 baciloscopias, em 3.781 (7,1 por cento) foram detectados a presença de BAAR com positividade 4,9 por cento para diagnóstico, 1,2 por cento para controle de tratamento e 1 por cento para o sem informação quanto ao quesito diagnóstico e ou controle de tratamento. Foram realizadas 7.603 (14,3 por cento) culturas, havendo isolamento de microbactérias em 1.134 (14,9 por cento) amostras, 13,2 por cento para diagnóstico, 1,4 por cento para controle de tratamento e 0,3 por cento para o sem informação. Foram identificadas 936 cepas pertencentes ao complexo Mycobacterium tuberculosis e 48 ao complexo M.avium. O perfil de sensibilidade de 381 cepas de M.tuberculosis demonstrou que 78,0 por cento foram sensíveis às drogas testadas e 22,0 por cento resistentes a pelo menos uma droga. As análises destes dados contribuem para as ações do Programa de Controle da Tuberculose, auxiliando na determinação do perfil epidemiológico da doença na regiäo de Campinas e nas DIRs envolvidas. (AU)

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With the purpose of evaluating the bacteriological diagnosis and the tuberculosis treatmentcontrol, the authors analyzed, in the years of 1999/2000, the tests processed by the Adolfo Lutz Institutein Campinas (SP), Health Regional Department (DIRS) and laboratories related to it from XII, XV and XX,witch serve 93 towns. During that period 53.341 samples were undertaken, 3.781 (7,1%) detected thepresence of BAAR with positiveness of 4,9% for diagnosis, 1,2% for treatment control and 1,0% for theones without information regarding the diagnosis item or treatment control. 7.603(14,3%) cultures weregrown, with isolation of micobacteria taking place in 1.134 (14,9%); 13,2% for diagnosis, 1,4% for treatmentcontrol and 0,3% for the ones lacking information. 936 heathers belonging to Mycobacterium tuberculosiscomplex and 48 belonging do M. avium complex were identified. The profile of sensitiveness of 381heathers of M. tuberculosis showed that 78% were sensitive to tested drugs used and 22% resisted atleast one drug. That data analyses have contributed to the realizations of the Tuberculosis ControlProgram, helping to determining the epidemiological profile of the illness in the Campinas region andinvolved DIRs. (AU)

Asunto(s)

Bacteriología , Tuberculosis Pulmonar , Resistencia a Medicamentos , Complejo Mycobacterium avium , Mycobacterium tuberculosis

RESUMEN

Tendo em vista que as dificuldades no diagnóstico laboratorial das men.ingites bacterianas poderiam estar ligadas a agentes antibacterianos presentes no líquido cefalorraquidiano (LCR), foi desenvolvida metodologia fácil para evidenciar a sua presença nesse material. Em placa de ágar Muellez Hinton, semeada com Staphylococcue aureus ATCC 6538 P não produtora de penicilinase e sensível a grande número de an tibacterianos, foi colocado um molde metálico com orifícios nos quais se aplicou o LCR que se difundia impedindo ou não o crescimento da bactéria-padrão. Das 641 amostras de LCR estudadas, em 38,53% o antibacteriano estava presente, e em 60,530/0 das amostras o agente etiológico foi caracterizado por exames bacteriológicos e/ou imunológicos. Os restantes 39,47% foram considerados de etiologia indeterminada. Pela análise estatística pode-se concluir que a presença de antibacteriano no LCR prejudica sobremaneira o diagnóstico laboratorial das meningites bacterianas. Considerando que a técnica empregada é de facil execução, o seu uso rotineiro contribuirá para melhor interpretação dos resultados, principalmente daquelas amostras de LCR purulento em que não se visualiza ou identifica o agente bacteriano (AU).

Asunto(s)

Humanos , Meningitis , Antibacterianos