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Acute respiratory infections remain a leading cause of death among young children in low- and middle-income countries. The etiological diagnosis of these infections is challenging due to the similarity in clinical presentations and overlapping symptoms caused by various pathogens. This database provides comprehensive epidemiological, clinical, paraclinical, and biological data on 801 Moroccan children admitted to the Children's Hospital of Rabat for the management of Clinical Severe Pneumonia. Identification of the pathogens responsible of respiratory infections was carried out using blood samples for hemoculture, standard bacterial culture and multiplex RT-PCR using the TrueScience RespiFinder Pathogen Identification Panel (Applied Biosystems).
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Bacteriophages (phages), viruses that infect bacteria, are found in abundance not only in the environment but also in the human body. The use of phages for the diagnosis of melioidosis, a tropical infectious disease caused by Burkholderia pseudomallei, is emerging as a promising novel approach, but our understanding of conditions under which Burkholderia prophages can be induced remains limited. Here, we first demonstrated the isolation of Burkholderia phages from the hemocultures of melioidosis patients. The B. pseudomallei-positive hemoculture bottles were filtered to remove bacteria, and then phages were isolated and purified by spot and double agar overlay plaque assays. Forty blood samples (hemoculture-confirmed melioidosis) were tested, and phages were found in 30% of the samples. Transmission electron microscopy and genome analysis of the isolated phages, vB_HM387 and vB_HM795, showed that both phages are Myoviruses. These two phages were stable at a pH of 5-7 and temperatures of 25-37°C, suggesting their ability to survive in human blood. The genome sizes of vB_HM387 and vB_HM795 are 36.3 and 44.0 kb, respectively. A phylogenetic analysis indicated that vB_HM387 has homologs, but vB_HM795 is a novel Myovirus, suggesting the heterogeneity of Burkholderia phages in melioidosis patients. The key finding that Burkholderia phages could be isolated from the blood of melioidosis patients highlights the potential application of phage-based assays by detecting phages in blood as a pathogen-derived biomarker of infection.
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BACKGROUND: Sepsis is a life-threatening condition for which critically important antimicrobials are often indicated. The value of blood culture for sepsis is indisputable, but appropriate guidelines on sampling and interpretation are currently lacking in cattle. OBJECTIVE: Compare the diagnostic accuracy of 2 blood culture media (pediatric plus [PP] and plus aerobic [PA]) and hypoglycemia for bacteremia detection. Estimate the contamination risk of blood cultures in critically ill calves. ANIMALS: One hundred twenty-six critically ill calves, 0 to 114 days. METHODS: Retrospective cross-sectional study in which the performance of PP, PA and hypoglycemia to diagnose sepsis was assessed using a Bayesian latent class model. A Cox proportional hazards model was used to compare time to positivity (TTP). Potential contamination was descriptively analyzed. Isolates were considered relevant when they were; member of the Enterobacterales, isolated from both blood cultures vials, or well-known, significant bovine pathogens. RESULTS: The sensitivities for PP, PA, and hypoglycemia were higher when excluding assumed contaminants; 68.7% (95% credibility interval = 30.5%-93.7%), 87.5% (47.0%-99.5%), and 61.3% (49.7%-72.4%), respectively. Specificity was estimated at 95.1% (82.2%-99.7%), 94.2% (80.7%-99.7%), and 72.4% (64.6%-79.6%), respectively. Out of 121 interpretable samples, 14.9% grew a presumed contaminant in PA, PP, or both. There was no significant difference in the TTP between PA and PP. CONCLUSIONS AND CLINICAL IMPORTANCE: PA and PP appear to outperform hypoglycemia as diagnostic tests for sepsis. PA seems most sensitive, but a larger sample size is required to verify this. Accuracy increased greatly after excluding assumed contaminants. The type of culture did not influence TTP or the contamination rate.
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Teorema de Bayes , Cultivo de Sangre , Enfermedades de los Bovinos , Medios de Cultivo , Hipoglucemia , Sensibilidad y Especificidad , Sepsis , Animales , Bovinos , Cultivo de Sangre/veterinaria , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/sangre , Hipoglucemia/veterinaria , Hipoglucemia/diagnóstico , Hipoglucemia/sangre , Estudios Retrospectivos , Sepsis/veterinaria , Sepsis/diagnóstico , Sepsis/microbiología , Estudios Transversales , Masculino , FemeninoRESUMEN
Abstract Routine blood culture is used for the detection of bloodstream infections by aerobic and anaerobic bacteria and by common pathogenic yeasts. A retrospective study was conducted in a public hospital in Maceió-AL, by collecting data of all medical records with positive blood cultures. Out of the 2,107 blood cultures performed, 17% were positive with Staphylococcus coagulase negative (51.14%), followed by Staphylococcus aureus (11.21%) and Klebsiella pneumoniae (6.32%). Gram-positive bacteria predominated among positive blood cultures, highlighting the group of Staphylococcus coagulase-negative. While Gram-negative bacteria had a higher number of species among positive blood cultures.
Resumo A cultura sanguínea de rotina é usada para a detecção de infecções na corrente sanguínea por bactérias aeróbias e anaeróbias e por leveduras patogênicas comuns. Estudo retrospectivo realizado em hospital público de Maceió-AL, por meio da coleta de dados de todos os prontuários com culturas sanguíneas positivas. Das 2.107 culturas sanguíneas realizadas, 17% foram positivas com Staphylococcus coagulase negativo (51,14%), seguido por Staphylococcus aureus (11,21%) e Klebsiella pneumoniae (6,32%). As bactérias Gram-positiva predominaram entre as culturas de sangue positivas, destacando-se o grupo das Staphylococcus coagulase-negativo. Enquanto as bactérias Gram-negativas apresentaram um número maior de espécies entre as culturas de sangue positivas.
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Abstract Routine blood culture is used for the detection of bloodstream infections by aerobic and anaerobic bacteria and by common pathogenic yeasts. A retrospective study was conducted in a public hospital in Maceió-AL, by collecting data of all medical records with positive blood cultures. Out of the 2,107 blood cultures performed, 17% were positive with Staphylococcus coagulase negative (51.14%), followed by Staphylococcus aureus (11.21%) and Klebsiella pneumoniae (6.32%). Gram-positive bacteria predominated among positive blood cultures, highlighting the group of Staphylococcus coagulase-negative. While Gram-negative bacteria had a higher number of species among positive blood cultures.
Resumo A cultura sanguínea de rotina é usada para a detecção de infecções na corrente sanguínea por bactérias aeróbias e anaeróbias e por leveduras patogênicas comuns. Estudo retrospectivo realizado em hospital público de Maceió-AL, por meio da coleta de dados de todos os prontuários com culturas sanguíneas positivas. Das 2.107 culturas sanguíneas realizadas, 17% foram positivas com Staphylococcus coagulase negativo (51,14%), seguido por Staphylococcus aureus (11,21%) e Klebsiella pneumoniae (6,32%). As bactérias Gram-positiva predominaram entre as culturas de sangue positivas, destacando-se o grupo das Staphylococcus coagulase-negativo. Enquanto as bactérias Gram-negativas apresentaram um número maior de espécies entre as culturas de sangue positivas.
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Humanos , Sepsis , Bacterias Gramnegativas , Brasil , Estudios Retrospectivos , HospitalesRESUMEN
Mixed infections by different Trypanosoma species or genotypes are a common and puzzling phenomenon. Therefore, it is critical to refine the diagnostic techniques and to understand to what extent these methods detect trypanosomes. We aimed to develop an accessible strategy to enhance the sensitivity of the hemoculture, as well as to understand the limitations of the hemoculture and the blood clot as a source of parasitic DNA. We investigated trypanosomatid infections in 472 bats by molecular characterization (18S rDNA gene) of the DNA obtained from the blood clot and, innovatively, from three hemoculture sample types: the amplified flagellates ("isolate"), the pellet of the culture harvested in its very initial growth stage ("first aliquot"), and the pellet of non-grown cultures with failure of amplification ("sediment"). We compared (a) the characterization of the flagellates obtained by first aliquots and isolates; and (b) the performance of the hemoculture and blood clot for trypanosomatid detection. We observed: (i) a putative new species of Bodo in Artibeus lituratus; (ii) the potential of Trypanosoma cruzi selection in the hemoculture; (iii) that the first aliquots and sediments overcome the selective pressure of the hemoculture; and (iv) that the blood clot technique performs better than the hemoculture. However, combining these methods enhances the detection of single and mixed infections.
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Bloodstream infection (BI) is the cause of high mortality. Hospital bloodstream infection (HBI) complicates hemodialysis, pneumonia, oncohematological diseases. Positive hemoculture obtaining depends on the volume of blood inoculation, the number of blood samples, the incubation time. To test the principles of microbiological culturomics in the diagnosis BI of hospital patients with a therapeutic profile. 848 hospital cardiac patients with suspected BI were included. 10 ml of blood were taken intravenously with a syringe, blood was inoculated into 200 ml of the heart-brain medium (HBM) in an anaerobic bottle. It was incubated for 7 or more days in a thermostat at +37º C. The hemocultures were obtained in 64.3% of cases with paired blood sampling with an interval of 30 minutes whereas an increase in the number of blood samples reduced the effectiveness of obtaining hemocultures to 9.1%. When incubating bottles for more than 7 days there were obtained 200 additional hemocultures containing 239 strains of microorganisms. Episodes of HBI were observed more often in the cases of the circulatory system (77.8%), including infectious endocarditis (IE) (47.0%), rheumatism (22.1%), myocarditis (14.6%). Episodes of HBI occurred more often in men with IE and coronary heart disease, in women - with rheumatism and myocarditis. Patients aged 45-75 were in the group of risk with a probability of complications of HBI up to 73.7%. When examining the blood of 848 hospital patients of cardiological profile HBI was detected in 38.3% of cases. Among clinical isolates gram-positive cocci with a great number S.epidermidis prevailed. Polymicrobial hemocultures (16.3%) were characterized by two and three associates in one blood sample. Among the hematological indicators in HBI there were: leukocytosis, increased ESR, lymphocytosis, decreased hemoglobin; increased values of fibrinogen, CRP, γ-globulin, α2-globulin, low levels of total protein and A/G coefficient. The techniques of microbiological culturomics were used. HBI was diagnosed in 38.3% of the therapeutic patients of cardiological profile. The etiology of HBI was characterized by polymicrobicity in 16.3% of cases. Hematological markers of HBI were identified.
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Bacteriemia , Miocarditis , Enfermedades Reumáticas , Sepsis , Femenino , Corazón , Hospitales , Humanos , Masculino , Sepsis/diagnósticoRESUMEN
OBJECTIVE: To reduce the number of blood culture samples collected. PATIENTS AND METHOD: We performed a cluster randomized controlled trial in adult acute care, and subacute care and rehabilitation wards in a university hospital in France. A poster associating an image of eyes looking at the reader with a summary of blood culture sampling guidelines was displayed in hospital wards in the intervention group. The incidence rate of blood cultures per 1000 days during pre- and post-intervention periods was calculated. RESULTS: Thirty-one wards participated in the study. The median difference in blood cultures/1000 days between periods was -1.863 [-11.941; 1.007] in the intervention group and -5.824 [-14.763; -2.217] in the control group (P=0.27). CONCLUSION: The intervention did not show the expected effect, possibly due to the choice of blood cultures as a target of good practice, but also to confounding factors such as the stringent policy of decreasing unnecessary costly testing.
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Cultivo de Sangre , Recolección de Muestras de Sangre/estadística & datos numéricos , Carteles como Asunto , HumanosRESUMEN
In cases of septic shock, high quality blood cultures are essential for identifying the causative agent and adapting the antibiotic treatment. Student nurses are relatively unfamiliar with recent guidelines on collecting blood cultures. Improving how they are taught during initial training, a key moment for the learning of best care practices, should help to optimise the global management of septic shock.
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Cultivo de Sangre , Educación en Enfermería , Estudiantes de Enfermería/psicología , Antibacterianos/uso terapéutico , Humanos , Aprendizaje , Choque Séptico/tratamiento farmacológico , Choque Séptico/enfermeríaRESUMEN
OBJECTIVE: Pathogens are usually identified from blood cultures using a two-step procedure: Gram staining on the day of bacterial growth (D0), followed by identification and susceptibility testing the following day (D1). We aimed to evaluate the use of rapid tests performed on D0 in patients presenting with Enterobacteriaceae bacteremia. PATIENTS AND METHODS: Patients with≥1 positive monomicrobial blood culture with Gram staining suggestive of an Enterobacteriaceae were prospectively included. Two successive strategies were evaluated: i) conventional strategy (CS), ii) combination of a rapid identification test and third-generation cephalosporin susceptibility testing (rapid strategy, e.g. RS). RESULTS: Eighty-three patients were included (CS=42; RS=41). Compared with CS, the median delay of identification was significantly shorter with RS (22 hours [20-27] vs. 47 hours [42-53]; P<0.001). Patients in the RS group more frequently received an effective (82.9% vs. 73.8%, P=0.43) and appropriate (70.7% vs. 54.7%, P=0.17) antibiotic therapy on D1. Moreover, all five RS patients infected with a non-susceptible strain received an effective therapy on D1 versus only three of eight CS patients. CONCLUSIONS: Use of rapid testing was associated with a reduced time to result availability. This strategy should be useful to initiate an early effective and appropriate therapy and to improve the care of patients.
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Bacteriemia/diagnóstico , Bacteriemia/terapia , Pruebas Diagnósticas de Rutina/métodos , Infecciones por Enterobacteriaceae/diagnóstico , Infecciones por Enterobacteriaceae/terapia , Anciano , Anciano de 80 o más Años , Bacteriemia/epidemiología , Bacteriemia/microbiología , Pruebas Diagnósticas de Rutina/estadística & datos numéricos , Intervención Médica Temprana/métodos , Enterobacteriaceae/aislamiento & purificación , Infecciones por Enterobacteriaceae/epidemiología , Infecciones por Enterobacteriaceae/microbiología , Femenino , Violeta de Genciana , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Fenazinas , Sepsis/tratamiento farmacológico , Sepsis/epidemiología , Sepsis/microbiología , Factores de Tiempo , Tiempo de Tratamiento/estadística & datos numéricosRESUMEN
Infective endocarditis is a grave disease because of a high level of morbidity and mortality. We conducted a retrospective analysis of 100 patients hospitalized for infective endocarditis from January 2009 until December 2015. The mean age was 41 years with a male predominance. Infective endocarditis occurs on a native valve (77 patients), prosthetic valves (12 patients), Pacemaker (7 patients) and congenital heart disease (4 patients). The diagnostic delay was 77 days on average. The fever was present in 85 patients. Blood cultures were negative in 54 patients. Echocardiography allowed visualizing the vegetations in 95 patients. In patients with prosthetics valves, a paraprosthetic regurgitation was diagnosed in 54 % of these patients and vegetation in 18 %. The main complications are heart failure (42), neurological (19), spleen (10) and renal (23) complications. The evolution under medical treatment was marked by the clinical-laboratory improvement in 57 patients, the lack of improvement with persistent fever in 21 patients. The mortality rate of our series is 23 %. Only seven patients underwent urgent surgery for hemodynamic and infectious indications. At the end of this work, it is emphasized that the epidemiological profile of infective endocarditis does not follow the changes of Wold. Its management is difficult in our context (significant diagnostic delay, often negative blood cultures, high level of morbidity, lack of recourse to early surgery). The improvement of this disease consistes of the prevention of acute articular rhumatism and the establishment of an heart team endocarditis to adapt international recommendations to our context.
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Endocarditis/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Cultivo de Sangre , Niño , Diagnóstico Tardío , Ecocardiografía , Endocarditis/diagnóstico , Endocarditis/mortalidad , Endocarditis/terapia , Endocarditis Bacteriana/microbiología , Femenino , Fiebre/etiología , Cardiopatías Congénitas/complicaciones , Insuficiencia Cardíaca/etiología , Humanos , Masculino , Persona de Mediana Edad , Marruecos , Marcapaso Artificial/efectos adversos , Infecciones Relacionadas con Prótesis/complicaciones , Estudios Retrospectivos , Enfermedades Reumáticas/prevención & control , Factores de Riesgo , Resultado del Tratamiento , Adulto JovenRESUMEN
Bacterial blood stream infections (BSI) are a common cause of mortality and morbidity globally. As the causative agents and the resulting treatment decisions vary, near-patient testing and surveillance tools are necessary to monitor bacterial causes and resistance to antimicrobial agents. The gold standard to identify BSIs is blood culture (BC), a methodology not widely available in resource-limited settings. The aim of the study was to map out a target product profile of a simplified BC system (SBCS) to inform product development efforts. To identify the desired characteristics of a SBCS, we enlisted a small group of specialists working in Africa and Asia. Questions were used to understand challenges and how these constraints inform system requirements. The specialists were infectious disease physicians, public health/clinical microbiologists, clinical researchers, and technology experts with different geographical backgrounds. All suggested that BC should ideally be available at the district hospital level. Many of the same operational challenges, such as limited availability of culture bottles, electricity and internet connectivity, profuse dust, the lack of ambient temperature control, and human capacity constraints were identified across the different regions. BCs, although the accepted gold standard for diagnosis of BSIs, are not widely available outside of reference/research centers in Africa and Asia. To extend the reach of this important tool, it is crucial to engage product developers and academic research partners to develop accessible alternatives.
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Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.
Resumo Trypanosoma (Megatrypanum) theileri é um protozoário flagelado que infecta ruminantes e apresenta alta diversidade genética. Neste estudo, investigamos as taxas de prevalência deste protozoário com base na hemocultura e no diagnóstico molecular. Os isolados de T . theileri obtidos foram caracterizados pelos marcadores moleculares SSU rDNA e gGAPDH e o diagnóstico molecular foi baseado no gene do tipo Catepsina L (PCR-TthCATL). O PCR-TthCATL e a hemocultura indicaram uma taxa de prevalência total de 8,13% e a sequência derivada do gene Catepsina L denominada IB de T. theileri foi identificada pela primeira vez em bovinos da Amazônia Ocidental, bem como a IF no Brasil. Também descrevemos uma possível nova sequência derivada da PCR-TthCATL em bovinos, designada IL.
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Animales , Femenino , Bovinos , Trypanosoma/clasificación , Tripanosomiasis Bovina/parasitología , Variación Genética/genética , Enfermedades de los Bovinos/parasitología , Filogenia , Trypanosoma/genética , Trypanosoma/inmunología , Tripanosomiasis Bovina/diagnóstico , Tripanosomiasis Bovina/epidemiología , Brasil/epidemiología , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Reacción en Cadena de la Polimerasa , ADN Protozoario/genética , Catepsina L/genética , GenotipoRESUMEN
Coagulase-negative staphylococci (CNS) are important pathogens causing nosocomial infections worldwide with increasing resistance to antimicrobials. The aim of this study was to characterize resistance aspects of CNS isolated from patients with bloodstream infections acquired in hospitals in Belo Horizonte, MG, Brazil. Staphylococcus strains were characterized using repetitive sequence-based polymerase chain reaction (rep-PCR) fingerprinting with (GTG)5 primer. Phenotypic resistance was analyzed using AST-P5085 card (bioMérieuxVitek®). PCR was used to detect mecA, vanA, blaZ, ermA/B/C, aac-aphD, and SCC-mec. For statistical analyses, we used hierarchical cluster, chi-square test (χ2), and correspondence. Several clusters were formed within the same species using (GTG)5 primer, and strains showed resistance to the following antimicrobials: benzylpenicillin (100%); oxacillin (93.1%); gentamicin (36.3%); ciprofloxacin (63.7%); moxifloxacin (32.7%); norfloxacin (81.0%); erythromycin (86.2%); clindamycin (75.8%); linezolid, teicoplanin and vancomycin (1.7%); tigecycline (0%); fusidic acid (10.35%); rifampicin (13.7%); and trimethoprim/sulfamethoxazole (46.5%). Regarding genotypic analyses, 40%, 0%, 78%, 42%, 100%, 24%, and 30% were positive for mecA, vanA, blaZ, ermA, ermB, ermC, and aac-aphD, respectively. Regarding staphylococcal cassette mec (SCCmec) type, 3.4% presented type I; 5.0% type II; 27.1% type III; 20.3% type IIIA; and 32.2% type IIIB. Six clusters were formed and frequency distributions of resistant strains to oxacillin, gentamicin, ciprofloxacin, moxifloxacin, norfloxacin, erythromycin, clindamycin, linezolid, teicoplanin, vancomycin, fusidic acid, rifampicin, and trimethoprim/sulfamethoxazole, and mecA, blaZ, ermC, aac-aphD, and SCCmec type differed (p < 0.001). In conclusion, the strains investigated in this study were multidrug resistant and carried multiple antibiotic resistance genes.
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Bacteriemia/microbiología , Coagulasa/genética , Farmacorresistencia Bacteriana Múltiple/genética , Infecciones Estafilocócicas/microbiología , Staphylococcus/genética , Staphylococcus/aislamiento & purificación , Antibacterianos/uso terapéutico , Bacteriemia/tratamiento farmacológico , Brasil , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Infecciones Estafilocócicas/tratamiento farmacológico , Staphylococcus/efectos de los fármacosRESUMEN
La bacteriemia es una complicación grave de las infecciones bacterianas. Un diagnóstico temprano del microorganismo responsable permite aplicar tratamientos efectivos en menor intervalo de tiempo. Los hemocultivos son diagnosticadores clínicos diseñado para este fin. Objetivo: Realizar un estudio de estabilidad acelerado de un lote del hemocultivo HemoCen Aerobio que permita planificar su diseño en estante en condiciones reales. Métodos: Se formuló un lote del hemocultivo HemoCen Aerobio en el Centro Nacional de Biopreparados, BioCen y se envasó asépticamente en los Laboratorios Biológicos Farmacéuticos, LABIOFAM. Se llevó a cabo un estudio de estabilidad acelerado por el Método de Arrenhius. Los frascos se conservaron durante 120 días a 15 °C, 30 °C y 50 °C. Se realizaron evaluaciones físico-químicas, organolépticas y capacidad de promoción de crecimiento de Staphylococcus aureus ATCC 25923 a los 7, 15, 30, 60 y 120 días. Resultados: El estudio de estabilidad demostró que el pH y el color del medio se deterioran progresivamente en el tiempo cuando las temperaturas aumentan entre 30 °C y 50 °C. La promoción de crecimiento de Staphylococcus aureus resultó favorable con índices de recuperación entre 20 y 40 UFC·frasco-1. Discusión: HemoCen Aerobio resulta funcional con un desempeño analítico satisfactorio, cuyos índices de recuperación microbiana se encuentran acorde a los valores reportados en bacteriemias de escasa magnitud. Estos resultados sientan las bases para planificar un estudio de estabilidad en estante en condiciones reales. Conclusión: Se estima un período de validez de 2 años(AU)
Bacteremia is a serious complication of bacterial infections. Early diagnosis of the causative organism allows applying appropriate treatments in a shorter time interval. Hemocultures are clinical diagnosticians designed for this purpose. Objective: Perform an accelerated stability study of a batch of HemoCen Aerobic hemoculture that allows planning its shelf designed in true conditions. Methods: A batch of HemoCen Aerobic hemoculture was formulated at the National Bioproducts Center, BioCen, and aseptically packaged at the Biological Pharmaceutical Laboratories, LABIOFAM. An accelerated stability study was carried out by the Arrenhius Method. The bottles were stored for 120 days at 15 °C, 30 °C and 50 °C. Physicochemical, organoleptic and growth promotion capacity evaluations of Staphylococcus aureus ATCC 25923 were realized at 7, 15, 30, 60 and 120 days. Results: The stability study demonstrated that the pH and the color of the medium progressively deteriorate over time as temperatures increase between 30 °C and 50 °C. Growth promotion of Staphylococcus aureus was favorable with recovery rates between 20 and 40 CFU bottle-1. Discussion: HemoCen Aerobic is functional with a satisfactory analytical performance, which recovery rates are consistent with the values reported in bacteremia of low magnitude. These results provide the basis for planning a shelf stability study under real conditions. Conclusion: A durability period of 2 years was estimated(AU)
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Humanos , Bacteriemia/diagnóstico , Diagnóstico Precoz , Cultivo de Sangre/métodosRESUMEN
Contaminated and infected wounds occur very frequently in veterinary medicine and can cause systemic inflammatory response syndrome, sepsis, and death. This study aimed to test the feasibility of collecting wound material by deep-tissue or punch biopsy for microbial culture, determine the frequency of bacteria in the wound(s) and blood cultures and the susceptibility of these microbes to antimicrobials, and evaluate clinical parameters that could be related to prognosis. Thirty dogs with wounds and signs of SIRS/sepsis were included in this study. Bacteria were isolated from all wounds and 41 bacterial isolates could be identified based on culture of the materials collected by punch biopsy; 53.66% of the isolates were gram-negative, mainly involving Pseudomonas aeruginosa, Klebsiella pneumoniae, and Enterococcus spp., and 46.34% were gram-positive bacteria such as Streptococcus spp., Enterococcus spp., and Staphylococcus spp. The survival rate was 66.67%. Based on blood culture analysis, we identified bacteremia in seven patients, predominantly of gram-negative bacteria, which negatively affected patient survival, as six dogs died. Hypoglycemia (≤60mg/dL) and severe hyperglycemia (≥180mg/dL) also negatively affected survival as 23.33% of the hypo/hyperglycemic dogs died. Factors such as blood lactate level at admission and hematocrit levels, and mean arterial pressure were not significantly correlated with death or survival of the dogs.(AU)
As feridas contaminadas e infectadas em cães ocorrem com grande frequência na medicina veterinária e podem causar síndrome da resposta inflamatória sistêmica, sepse e morte. Os objetivos do presente trabalho foram verificar a viabilidade da técnica de coleta de material da ferida por biópsia para realização de cultura microbiana, determinar a frequência das bactérias nas culturas das feridas e hemoculturas e a susceptibilidade destes agentes aos antimicrobianos, bem como avaliar parâmetros clínicos que pudessem ser relacionados ao prognóstico em 30 cães com feridas e sinais de SIRS/sepse. Foram isoladas bactérias de todas as feridas e a técnica de coleta de material para cultura por biópsia permitiu a obtenção de 41 agentes microbianos, sendo isoladas 53,66% bactérias Gram negativas e 46,34% Gram positivas, principalmente Pseudomonas aeruginosa, Klebsiella pneumoniae e Enterococcus spp. As bactérias gram positivas isoladas foram Streptococcus spp., Enterococcus spp. e Staphylococcus spp. A taxa de sobrevivência foi 66,67%. Na hemocultura constatou-se bacteremia em sete pacientes, com predominância de bactérias Gram negativas, o que influenciou negativamente na sobrevivência dos pacientes, pois seis cães vieram a óbito. A hipoglicemia (≤60mg/dL) ou hiperglicemia severa (≥180mg/dL), também influenciaram negativamente a sobrevivência, pois 23,33% dos pacientes hipo/hiperglicêmicos vieram a óbito. Já fatores como nível sérico de lactato na admissão do paciente, pressão arterial média (PAM) e hematócrito não apresentaram correlação estatística com o óbito ou sobrevivência destes pacientes.(AU)
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Animales , Perros , Infección de Heridas/complicaciones , Bacteriemia/veterinaria , Sepsis/veterinaria , Cultivo de Sangre/veterinaria , Técnicas Citológicas/veterinariaRESUMEN
Escherichia coli is the most common cause of bloodstream infections and community-acquired sepsis. The main aim of this study was to determine virulence characteristics of E. coli isolates from hemocultures of patients with a primary disease of urogenital tract, digestive system, a neoplastic blood disease, or other conditions. Results from a set of 314 E. coli isolates from hemocultures were compared to data from a previously published analysis of 1283 fecal commensal E. coli isolates. Genetic profiling of the 314 E. coli isolates involved determination of phylogenetic group (A, B1, B2, D, C, E, and F), identification of 21 virulence factors, as well as 30 bacteriocin-encoding determinants. Pulsed-field gel electrophoresis was used to analyze clonal character of the hemoculture-derived isolates. The E. coli isolates from hemocultures belonged mainly to phylogenetic groups B2 (59.9%) and D (21.0%), and less frequently to phylogroups A (10.2%) and B1 (5.7%). Commonly detected virulence factors included adhesins (fimA 92.0%, pap 47.1%, and sfa 26.8%), and iron-uptake encoding genes (fyuA 87.9%, fepC 79.6%, aer 70.7%, iucC 68.2%, and ireA 13.7%), followed by colibactin (pks island 31.5%), and cytotoxic necrotizing factor (cnf1 11.1%). A higher frequency of microcin producers (and microcin M determinant) and a lower frequency of colicin Ib and microcin B17 was found in hemoculture-derived isolates compared to commensal fecal isolates. E. coli isolates from hemocultures harbored more virulence genes compared to fecal E. coli isolates. In addition, hemoculture E. coli isolates from patients with primary diagnosis related to urogenital tract were clearly different and more virulence genes were detected in these isolates compared to both fecal isolates and hemoculture-derived isolates from patients with blood and gastrointestinal diseases.
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Infecciones por Escherichia coli/microbiología , Escherichia coli/aislamiento & purificación , Escherichia coli/patogenicidad , Sepsis/microbiología , Factores de Virulencia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Colicinas/genética , Electroforesis en Gel de Campo Pulsado , Escherichia coli/clasificación , Escherichia coli/genética , Femenino , Gastroenteritis/complicaciones , Genotipo , Humanos , Lactante , Masculino , Persona de Mediana Edad , Neoplasias/complicaciones , Filogenia , Infecciones Urinarias/complicaciones , Adulto JovenRESUMEN
BACKGROUND: False-positive blood cultures can lead to unnecessary risks and misuse of antibiotics; to reduce rates of false-positives, it would be useful to determine whether use of an antiseptic with a prolonged effect is required. METHODS: Clinical study of efficacy (blinded and randomized) to compare the rate of blood culture contamination when skin antisepsis was performed with 70% isopropyl alcohol or 2% chlorhexidine gluconate in 70% isopropyl alcohol in 2 hospitals. Patients aged 16 years or older with suspected bloodstream infection who were allocated in the emergency room, internal medicine ward, or intensive care unit were included. RESULTS: Five of 563 (0.9%) blood cultures from the isopropyl arm and 10 of 539 (1.9%) from the chlorhexidine arm were contaminated. No significant differences were observed among the rate of contamination (χ2=1.27; P = .3) or the relative risk of contamination (relative risk = 2.09; 95% confidence interval, 0.72-6.07; P = .2). CONCLUSIONS: The rates of blood contamination were not different when isopropyl alcohol and chlorhexidine were compared. Isopropyl alcohol could be used for skin antisepsis before blood collection.
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2-Propanol/administración & dosificación , Antiinfecciosos Locales/administración & dosificación , Cultivo de Sangre/métodos , Clorhexidina/administración & dosificación , Desinfección/métodos , Manejo de Especímenes/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Distribución Aleatoria , Adulto JovenRESUMEN
Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.
Resumo Trypanosoma (Megatrypanum) theileri é um protozoário flagelado que infecta ruminantes e apresenta alta diversidade genética. Neste estudo, investigamos as taxas de prevalência deste protozoário com base na hemocultura e no diagnóstico molecular. Os isolados de T . theileri obtidos foram caracterizados pelos marcadores moleculares SSU rDNA e gGAPDH e o diagnóstico molecular foi baseado no gene do tipo Catepsina L (PCR-TthCATL). O PCR-TthCATL e a hemocultura indicaram uma taxa de prevalência total de 8,13% e a sequência derivada do gene Catepsina L denominada IB de T. theileri foi identificada pela primeira vez em bovinos da Amazônia Ocidental, bem como a IF no Brasil. Também descrevemos uma possível nova sequência derivada da PCR-TthCATL em bovinos, designada IL.
RESUMEN
Coagulase-negative staphylococci (CNS) represent one of the most prevalent microorganisms in nosocomial infections worldwide, nevertheless little is known about their pathogenicity features. Thus, our aim was to characterize virulence aspects of CNS isolated from patients with bloodstream infections assisted in hospitals of Belo Horizonte, MG, Brazil. Strains were identified using bioMérieuxVitek® and for biofilm production evaluation, Congo Red Agar (CRA) and polystyrene plates were used. PCR was applied to detect icaA, icaB, icaC, atlE, sea, sec, sed, tsst-1 and agr. For statistical analyses were used hierarchical cluster, chi-square test and correspondence. 59 strains were analyzed, being S. haemolyticus the most prevalent. On CRA, 96.5% were biofilm producer, whereas on polystyrene plate, 100% showed adhesion at different times evaluated. Regarding genotypic analyses, 15.2%, 38.9%, 8.4%, 49.1%, 76.2%, 23.7%, 1.6%, 30.5% and 38.9% were positive for icaA, icaB, icaC, atlE, sea, sec, sed, tsst-1 and agr, respectively. Six clusters were formed and frequency distributions of agr, atlE, icaA, icaB, sea, sec, tsst-1 differed (P < 0.001). In conclusion, all strains were biofilm producer, with high prevalence of atlE, and had potential of toxin production, with high prevalence of sea. According to the group-analyses, icaB showed relationship with the strong adherence in samples.