Genomic Characterization of Campylobacter jejuni Associated with Perimyocarditis: A Family Case Report.

Campylobacter spp. is the leading cause of foodborne gastrointestinal infections in humans worldwide. This study reports the first case of four family members who had contact with the same source of Campylobacter jejuni contamination with different results. Only the little siblings were infected by the same C. jejuni strain, but with different symptoms. Whereas the daughter was slightly affected with mild enteritis, the son suffered a longer campylobacteriosis followed with a perimyocarditis. This is the first case of the youngest patient affected by C. jejuni-related perimyocarditis published to date. The genomes of both strains were characterized by whole-genome sequencing and compared with the C. jejuni NCTC 11168 genome to gain insights into the molecular features that may be associated with perimyocarditis. Various comparison tools were used for the comparative genomics analysis, including the identification of virulence and antimicrobial resistance genes, phase variable (PV) genes, and single nucleotide polymorphisms (SNPs) identification. Comparisons of the strains identified 16 SNPs between them, which constituted small but significant changes mainly affecting the ON/OFF state of PV genes after passing through both hosts. These results suggest that PV occurs during human colonization, which modulates bacteria virulence through human host adaptation, which ultimately is related to complications after a campylobacteriosis episode depending on the host status. The findings highlight the importance of the relation between host and pathogen in severe complications of Campylobacter infections.

A Pilot Study for the Evaluation of an Interferon Gamma Release Assay (IGRA) To Measure T-Cell Immune Responses after SARS-CoV-2 Infection or Vaccination in a Unique Cloistered Cohort.

Assessment of T-cell responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens may be of value to determine long-lasting protection to breakthrough infections or reinfections. Interferon gamma release assay is a validated method to test cellular immunity in mycobacterial infections and has been proposed for patients with SARS-CoV-2 infection or vaccination. Quantitative IgG to spike and qualitative IgG to nucleocapsid antigens were determined by chemiluminescence microparticle immunoassay using the Architect platform (Abbott), and interferon gamma release assays against two Qiagen proprietary mixes of SARS-CoV-2 spike protein (antigen 1 and antigen 2) were performed for a selected group of subjects. A total of 121 subjects in a cloistered institution after a COVID-19 outbreak was studied. IgG spike levels and interferon gamma concentrations were highest among subjects after two doses of vaccine, followed by patients with a longer history of past COVID-19 and no vaccination. The best cutoff for the interferon gamma assay was 25 IU/L for all subgroups of individuals and the two sets of SARS-CoV-2 antigens studied. Testing T-cell response may be of clinical utility to determine immunity after exposure to SARS-CoV-2 antigens, with the interferon gamma concentration of 25 IU/L as the best cutoff either after infection or vaccination.

Viral RNA load in plasma is associated with critical illness and a dysregulated host response in COVID-19.

BACKGROUND: COVID-19 can course with respiratory and extrapulmonary disease. SARS-CoV-2 RNA is detected in respiratory samples but also in blood, stool and urine. Severe COVID-19 is characterized by a dysregulated host response to this virus. We studied whether viral RNAemia or viral RNA load in plasma is associated with severe COVID-19 and also to this dysregulated response. METHODS: A total of 250 patients with COVID-19 were recruited (50 outpatients, 100 hospitalized ward patients and 100 critically ill). Viral RNA detection and quantification in plasma was performed using droplet digital PCR, targeting the N1 and N2 regions of the SARS-CoV-2 nucleoprotein gene. The association between SARS-CoV-2 RNAemia and viral RNA load in plasma with severity was evaluated by multivariate logistic regression. Correlations between viral RNA load and biomarkers evidencing dysregulation of host response were evaluated by calculating the Spearman correlation coefficients. RESULTS: The frequency of viral RNAemia was higher in the critically ill patients (78%) compared to ward patients (27%) and outpatients (2%) (p < 0.001). Critical patients had higher viral RNA loads in plasma than non-critically ill patients, with non-survivors showing the highest values. When outpatients and ward patients were compared, viral RNAemia did not show significant associations in the multivariate analysis. In contrast, when ward patients were compared with ICU patients, both viral RNAemia and viral RNA load in plasma were associated with critical illness (OR [CI 95%], p): RNAemia (3.92 [1.183-12.968], 0.025), viral RNA load (N1) (1.962 [1.244-3.096], 0.004); viral RNA load (N2) (2.229 [1.382-3.595], 0.001). Viral RNA load in plasma correlated with higher levels of chemokines (CXCL10, CCL2), biomarkers indicative of a systemic inflammatory response (IL-6, CRP, ferritin), activation of NK cells (IL-15), endothelial dysfunction (VCAM-1, angiopoietin-2, ICAM-1), coagulation activation (D-Dimer and INR), tissue damage (LDH, GPT), neutrophil response (neutrophils counts, myeloperoxidase, GM-CSF) and immunodepression (PD-L1, IL-10, lymphopenia and monocytopenia). CONCLUSIONS: SARS-CoV-2 RNAemia and viral RNA load in plasma are associated with critical illness in COVID-19. Viral RNA load in plasma correlates with key signatures of dysregulated host responses, suggesting a major role of uncontrolled viral replication in the pathogenesis of this disease.

Molecular epidemiology of an enterovirus A71 outbreak associated with severe neurological disease, Spain, 2016.

IntroductionEnterovirus A71 (EV-A71) is an emerging pathogen that causes a wide range of disorders including severe neurological manifestations. In the past 20 years, this virus has been associated with large outbreaks of hand, foot and mouth disease with neurological complications in the Asia-Pacific region, while in Europe mainly sporadic cases have been reported. In spring 2016, however, an EV-A71 outbreak associated with severe neurological cases was reported in Catalonia and spread further to other Spanish regions.AimOur objective was to investigate the epidemiology and clinical characteristics of the outbreak.MethodsWe carried out a retrospective study which included 233 EV-A71-positive samples collected during 2016 from hospitalised patients. We analysed the clinical manifestations associated with EV-A71 infections and performed phylogenetic analyses of the 3'-VP1 and 3Dpol regions from all Spanish strains and a set of EV-A71 from other countries.ResultsMost EV-A71 infections were reported in children (mean age: 2.6 years) and the highest incidence was between May and July 2016 (83%). Most isolates (218/233) were classified as subgenogroup C1 and 217 of them were grouped in one cluster phylogenetically related to a new recombinant variant strain associated with severe neurological diseases in Germany and France in 2015 and 2016. Moreover, we found a clear association of EV-A71-C1 infection with severe neurological disorders, brainstem encephalitis being the most commonly reported.ConclusionAn emerging recombinant variant of EV-A71-C1 was responsible for the large outbreak in 2016 in Spain that was associated with many severe neurological cases.

Comparison of epidemiology and clinical characteristics of infections by human parechovirus vs. those by enterovirus during the first month of life.

UNLABELLED: Human parechoviruses (HPeV) have been recently recognized as important viral agents in paediatric infections. The aims of this study were to investigate the HPeV infection prevalence in infants <1 month in Spain and, secondly, to analyse the clinical and epidemiological characteristics of the infected patients compared with those infected by enterovirus (EV). Infants <1 month with neurological or systemic symptoms were included in a multicentre prospective study. EV and HPeV detection by RT-PCR and genotyping were performed in cerebrospinal fluids (CSF), sera or throat swabs. Out of the total of 84 infants studied during 2013, 32 were EV positive (38 %) and 9 HPeV positive (11 %). HPeV-3 was identified in eight cases and HPeV-5 in one. Mean age of HPeV-positive patients was 18 days. Diagnoses were fever without source (FWS) (67 %), clinical sepsis (22 %) and encephalitis (11 %). Leukocytes in blood and CSF were normal. Pleocytosis (p = 0.03) and meningitis (p = 0.001) were significantly more frequent in patients with EV infections than with HPeV. CONCLUSIONS: Although HPeV-3 infections were detected less frequently than EV, they still account for approximately 10 % of the cases analysed in infants younger than 1 month. HPeV-3 was mainly associated with FWS and without leukocytosis and pleocytosis in CSF. In these cases, HPeV screening is desirable to identify the aetiologic agent and prevent unnecessary treatment and prolonged hospitalization.

Surveillance for Enteroviruses Associated with Hand, Foot, and Mouth Disease, and Other Mucocutaneous Symptoms in Spain, 2006-2020.

Hand, foot, and mouth disease (HFMD) is a mild illness caused by enteroviruses (EV), although in some Asian countries, large outbreaks have been reported in the last 25 years, with a considerable incidence of neurological complications. This study describes epidemiological and clinical characteristics of EV infections involved in HFMD and other mucocutaneous symptoms from 2006 to 2020 in Spain. EV-positive samples from 368 patients were included. EV species A were identified in 85.1% of those typed EV. Coxsackievirus (CV) A6 was the prevalent serotype (60.9%), followed by EV-A71 (9.9%) and CVA16 (7.7%). Infections affected children (1-6 years old) mainly, and show seasonality with peaks in spring-summer and autumn. Clinical data indicated few cases of atypical HFMD as well as those with neurological complications (associated with the 2016 EV-A71 outbreak). Phylogenetic analysis of CVA6 VP1 sequences showed different sub-clusters circulating from 2010 to present. In conclusion, HFMD or exanthemas case reporting has increased in Spain in recent years, probably associated with an increase in circulation of CVA6, although they did not seem to show greater severity. However, EV surveillance in mucocutaneous manifestations should be improved to identify the emergence of new types or variants causing outbreaks and more severe pathologies.

[Drug-resistance in Mycobacterium tuberculosis in Castilla y León, Spain, 2001-2005: third collaborative study]. / Resistencias a fármacos de Mycobacterium tuberculosis en la Comunidad de Castilla y León (España), 2001-2005: tercer estudio multicéntrico.

INTRODUCTION: During 2001-2005, a regional anti-tuberculosis drug resistance survey was conducted in Castilla y León, Spain, in newly treated HIV negative tuberculosis (TB) patients. METHODS: A total of 918 Mycobacterium tuberculosis strains were studied (one strain per patient) from six hospitals corresponding to 46.7% of the total population of Castilla y León, using the proportion method on solid medium. RESULTS: Primary drug resistance was 4.2% (streptomycin 1.2%, isoniazid 3.2%, rifampin 0.3%, ethambutol 0.1% and pyrazinamide 0.5%). Mono-resistance was observed in 24 (2.6%) and resistance to both isoniazid and rifampin (multi-drug resistance) was detected in one case (0.1%). These results were not statistically significant compared to previous studies in the same Community. CONCLUSION: The incidence of primary drug resistance in the surveyed area was low, including isoniazid, allowing new anti-tuberculosis treatment with the standardised three-drug regimen to be started. Regular surveillance of drug resistance is recommended by the TB control programme in representative patient populations to optimize treatment regimens.

[Epidemiological and clinical characteristics of infants admitted to hospital due to human parechovirus infections: A prospective study in Spain]. / Características epidemiológicas y clínicas de los lactantes hospitalizados por infecciones por parechovirus humanos. Estudio prospectivo en España.

INTRODUCTION: Human parechovirus (HPeV) is one of the recently described picornaviridae viruses that have been associated with fever of unknown origin (FUO), clinical sepsis, gastroenteritis, meningitis, or encephalitis in very young infants. The aim of this study is to describe the epidemiology and clinical features of these viruses. PATIENTS AND METHODS: A prospective multicentre 3-year study was conducted in 12 hospitals in Spain. Out of 850 specimens examined, 47 were positive (5.52%), with HPeV-3 being the most frequent (29 cases). Infections occurred throughout the year, but mainly in May and July, and a biennial distribution was observed. More than half (57%) were neonates, and only 2 children were older than 3 months. Fever was present in all children, with irritability in 45%, rash in 18.6%, and diarrhoea in 14%. The results of biochemical tests were all in normal range. The most common final diagnosis was FUO (61%), followed by clinical sepsis (29%). Up to 29% of infants were admitted to the intensive care unit, but only one patient had sequelae. RESULTS: Out of 850 specimens examined, 47 were positive (5.52%) for HPeV, with HPeV-3 being the most frequent (29 cases). Infections occurred throughout the year, but mainly in May and July, and a biennial distribution was observed. More than half (57%) were neonates, and only 2 children were older than 3 months. Fever was present in all children, with irritability in 45%, rash in 18.6%, and diarrhoea in 14%. The results of biochemical tests were all in normal range. The most common final diagnosis was FUO (61%), followed by clinical sepsis (29%). Up to 29% of infants were admitted to the intensive care unit, but only one patient had sequelae CONCLUSIONS: HPeV circulates in our country, mainly during spring and summer, and affects young infants with a FUO and clinical sepsis. Molecular diagnostic techniques in all hospitals could help in improving the management of patients with these infections.

Perinatal infection and hypoxic-ischemic encephalopathy: a pilot study.

Recent studies suggest a synergic effect of infection and hypoxia-ischemia in the causation of perinatal brain damage. We conducted a prospective pilot study on the presence of infection in hypoxic-ischemic encephalopathy (HIE), focusing on neurotropic viruses. Sixteen newborns with HIE were included in the study. There were no confirmed cases of viral infection. There was a case of bacterial early onset sepsis and four cases of suspected sepsis due to clinical and/or analytical signs, but with negative cultures. Our results do not support universal screening for viral infection in cases of HIE.

Human parechovius and enterovirus in neonates: Distinct infections with overlapping features.

INTRODUCTION: Human parechoviruses (HPeVs) have recently been added to the family Picornaviridae, where Enteroviruses (HEV) belong. The specific characteristics of HPeV infection in the neonate are not clear, and their involvement in neonatal infection is believed to be largely underestimated. HPeV type 3 has been recently linked to sepsis-like illness and neurological involvement in the newborn. OBJECTIVE: To assess the involvement of HPeV in central nervous system (CNS) infections throughout the neonatal period in term newborns, describe their clinical, analytical and cerebrospinal fluid (CSF) characteristics, and compare them to HEV infections. METHODS: Term newborns admitted for neurological symptoms or a suspected infection, aged 0-30 days were prospectively recruited (September 2012-August 2014). Bacterial cultures were performed in all patients. Viral tests were performed in CSF including: RT-PCR for cytomegalovirus, Herpes simplex type 1 and 2, Epstein-Barr virus, HEV and HPeV. HEV and HPeV positive samples were genotyped. RESULTS: Fifty-seven newborns were diagnosed of sepsis-like illness and/or CNS alteration. HEV (8.7%) and HPeV-3 (3.5%) were the two most common viral agents involved during the study period. The most frequent symptom at admission was fever. Irritability was present in 1/2 of HPeV and 1/5 of HEV cases. There were no other neurological symptoms. Blood and CSF analysis were unremarkable in HPeV infections. All cases resolved favorably. CONCLUSIONS: HPeV infection was clinically very similar to that of HEV, while it featured normal blood and CSF analysis. PRACTICE IMPLICATIONS: HPeV should be considered by clinicians in the differential diagnosis of neonatal infection, particularly when blood and CSF analysis are unremarkable.

Características epidemiológicas y clínicas de los lactantes hospitalizados por infecciones por parechovirus humanos. Estudio prospectivo en España / Epidemiological and clinical characteristics of infants admitted to hospital due to human parechovirus infections: A prospective study in Spain

Introducción: Los parechovirus humanos (HPeV) son virus de la familia Picornaviridae, recientemente descritos, a los que se atribuyen cuadros de fiebre sin foco (FSF), sepsis clínica, gastroenteritis, meningitis o encefalitis fundamentalmente en lactantes pequeños. Nuestro objetivo fue describir la epidemiología y las características clínicas de las infecciones por HPeV en nuestro medio. Pacientes y métodos: Estudio multicéntrico prospectivo, llevado a cabo en 12 hospitales a nivel nacional, entre 2013-2015, en niños < 3 años con FSF, sepsis clínica o patología neurológica. Se realizó determinación de HPeV mediante RT-PCR en el Centro Nacional de Microbiología en suero, heces o líquido cefalorraquídeo. Resultados: Se analizan 47 infecciones por HPeV de un total de 850 muestras (5,52%), siendo HPeV-3 el más frecuente (29 casos), con predominio en mayo y julio, con una distribución bienal. El 57% eran neonatos y solo 2 > 3 meses. Todos los pacientes presentaron fiebre, el 45% irritabilidad, el 18,6% exantema y el 14% diarrea. No se observa ninguna alteración específica en las pruebas bioquímicas. El diagnóstico final más frecuente fue FSF (61%) seguido de sepsis clínica (29%). Aunque un 29% de los niños precisaron ingreso en cuidados intensivos, solo un paciente presentó secuelas. Conclusiones: Los HPeV circulan en nuestro país, afectando fundamentalmente a lactantes < 2 meses y se asocian a FSF y sepsis clínica, con un predominio en primavera y verano. Sería de interés implementar las técnicas moleculares de diagnóstico en todos los hospitales para reconocer y manejar adecuadamente estas infecciones (AU)

Introduction: Human parechovirus (HPeV) is one of the recently described picornaviridae viruses that have been associated with fever of unknown origin (FUO), clinical sepsis, gastroenteritis, meningitis, or encephalitis in very young infants. The aim of this study is to describe the epidemiology and clinical features of these viruses. Patients and methods: A prospective multicentre 3-year study was conducted in 12 hospitals in Spain. Out of 850 specimens examined, 47 were positive (5.52%), with HPeV-3 being the most frequent (29 cases). Infections occurred throughout the year, but mainly in May and July, and a biennial distribution was observed. More than half (57%) were neonates, and only 2 children were older than 3 months. Fever was present in all children, with irritability in 45%, rash in 18.6%, and diarrhoea in 14%. The results of biochemical tests were all in normal range. The most common final diagnosis was FUO (61%), followed by clinical sepsis (29%). Up to 29% of infants were admitted to the intensive care unit, but only one patient had sequelae. Results: Out of 850 specimens examined, 47 were positive (5.52%) for HPeV, with HPeV-3 being the most frequent (29 cases). Infections occurred throughout the year, but mainly in May and July, and a biennial distribution was observed. More than half (57%) were neonates, and only 2 children were older than 3 months. Fever was present in all children, with irritability in 45%, rash in 18.6%, and diarrhoea in 14%. The results of biochemical tests were all in normal range. The most common final diagnosis was FUO (61%), followed by clinical sepsis (29%). Up to 29% of infants were admitted to the intensive care unit, but only one patient had sequelae. Conclusions: HPeV circulates in our country, mainly during spring and summer, and affects young infants with a FUO and clinical sepsis. Molecular diagnostic techniques in all hospitals could help in improving the management of patients with these infections (AU)

Resistencias a fármacos de Mycobacterium tuberculosis en la Comunidad de Castilla y León (España), 2001–2005: tercer estudio multicéntrico / Drug-resistance in Mycobacterium tuberculosis in Castilla y León, Spain, 2001–2005: Third collaborative study

Introducción Durante el período 2001–2005 se llevó a cabo el tercer estudio de resistencia a los fármacos antituberculosos de primera línea en Castilla y León, España, en casos nuevos de tuberculosis (TB).Métodos Se estudiaron 918 cepas de Mycobacterium tuberculosis procedentes de pacientes virus de la inmunodeficiencia humana negativos correspondientes a 6 hospitales que atienden al 46,7% del total de la población de la comunidad, y se utilizó el método de las proporciones en medio sólido. Resultados La proporción de resistencias (resistencia primaria) fue del 4,2% y la resistencia simple (monorresistencia) fue del 2,6%. El 1,2% fueron resistentes a estreptomicina; el 3,2% a isoniacida (I); el 0,3% a rifampicina (R); el 0,1% a etambutol, y el 0,5% a pirazinamida, y se observó una cepa (0,1%) multirresistente (resistencia a I y a R). No hubo diferencias significativas con los estudios previos. Conclusiones La incidencia de resistencia primaria y de multirresistencia en la Comunidad de Castilla y León es baja, asimismo, la resistencia a I es aceptable. Consecuentemente, la pauta de tratamiento de la TB puede incluir solamente a los 3 fármacos (R, I y etambutol). Es conveniente, de acuerdo con los programas de control de la TB, realizar estudios de resistencia a los fármacos antituberculosos para optimizar las pautas de tratamiento (AU)

Introduction During 2001–2005, a regional anti-tuberculosis drug resistance survey was conducted in Castilla y León, Spain, in newly treated HIV negative tuberculosis (TB) patients. Methods A total of 918 Mycobacterium tuberculosis strains were studied (one strain per patient) from six hospitals corresponding to 46.7% of the total population of Castilla y León, using the proportion method on solid medium. Results Primary drug resistance was 4.2% (streptomycin 1.2%, isoniazid 3.2%, rifampin 0.3%, ethambutol 0.1% and pyrazinamide 0.5%). Mono-resistance was observed in 24 (2.6%) and resistance to both isoniazid and rifampin (multi-drug resistance) was detected in one case (0.1%). These results were not statistically significant compared to previous studies in the same Community. Conclusion The incidence of primary drug resistance in the surveyed area was low, including isoniazid, allowing new anti-tuberculosis treatment with the standardised three-drug regimen to be started. Regular surveillance of drug resistance is recommended by the TB control programme in representative patient populations to optimize treatment regimens (AU)